The invention relates to B7 homology 3 protein (B7-H3) antibodies and antibody drug conjugates (ADCs), including compositions and methods of using said antibodies and ADCs.

Description

68 parts

›RELATED APPLICATIONS

This application claims priority to U.S. Provisional Application No. 62/347,476, filed on Jun. 8, 2016, and to U.S. Provisional Application No. 62/366,511, filed on Jul. 25, 2016. The entire contents of the foregoing applications are expressly incorporated herein by reference.

›SEQUENCE LISTING

The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Jun. 7, 2017, is named 117813-12603_ST25.txt and is 159,744 bytes in size.

›BACKGROUND OF THE INVENTION

The B7 homology 3 protein (B7-H3) (also known as CD276 and B7RP-2, and referred to herein as “B7-H3”) is a type I transmembrane glycoprotein of the immunoglobulin superfamily. Human B7-H3 contains a putative signal peptide, V-like and C-like Ig domains, a transmembrane region and a cytoplasmic domain. Exon duplication in humans results in the expression of two B7-H3 isoforms having either a single IgV-IgC-like domain (2IgB7-H3 isoform) or a IgV-IgC-IgV-IgC-like domain (4IgB7-H3 isoform) containing several conserved cysteine residues. The predominant B7-H3 isoform in human tissues and cell lines is the 4IgB7-H3 isoform (Steinberger et al., J. Immunol. 172(4): 2352-9 (2004)).

B7-H3 has been reported as having both co-stimulatory and co-inhibitory signaling functions (see, e.g., Chapoval et al., Nat. Immunol. 2: 269-74 (2001); Suh et al., Nat. Immunol. 4: 899-906 (2003); Prasad et al., J. Immunol. 173: 2500-6 (2004); and Wang et al., Eur. J. Immunol. 35: 428-38 (2005)). For example, in vitro studies have shown B7-H3's co-stimulatory function since B7-H3 was able to increase proliferation of cytotoxic T-lymphocytes (CTLs) and upregulate interferon gamma (IFN-γ) production in the presence of anti-CD3 antibody to mimic the T cell receptor signal (Chapoval et al., 2001). Moreover, in vivo studies using cardiac allografts in B7-H3−/− mice showed decreased production of key cytokine, chemokine and chemokine receptor mRNA transcripts (e.g., IL-2, IFN-γ, monocyte chemoattractant protein (MCP-1) and IFN-inducible protein (IP)-10) as compared to wild-type control (Wang et al., 2005). In contrast, B7-H3 co-inhibitory function has been observed, for example, in mice where B7-H3 protein inhibited T-cell activation and effector cytokine production (Suh et al., 2003). Although no ligands have been identified for human B7-H3, murine B7-H3 has been found to bind to the triggering receptor expressed on myeloid cells (TREM-) like transcript 2 (TLT-2), a modulator of adaptive an innate immunity cellular responses. Binding of murine B7-H3 to TLT-2 on CD8 + T-cells enhances T-cell effector functions such as proliferation, cytotoxicity and cytokine production (Hashiguchi et al., Proc. Nat'l. Acad. Sci. U.S.A. 105(30): 10495-500 (2008)).

B7-H3 is not constitutively expressed in many immune cells (e.g., natural killer (NK) cells, T-cells, and antigen-presenting cells (APCs)), however, its expression can be induced. Further, the expression of B7-H3 is not restricted to immune cells. B7-H3 transcripts are expressed in a variety of human tissues including colon, heart, liver, placenta, prostate, small intestine, testis, and uterus, as well as osteoblasts, fibroblasts, epithelial cells, and other cells of non-lymphoid lineage, potentially indicating immunological and non-immunological functions (Nygren et al. Front Biosci. 3:989-93 (2011)). However, protein expression in normal tissue is typically maintained at a low level and thus, may be subject to post-transcriptional regulation.

B7-H3 is also expressed in a variety of human cancers, including prostate cancer, clear cell renal cell carcinoma, glioma, melanoma, lung cancer, non-small cell lung cancer (NSCLC), small cell lung cancer, pancreatic cancer, gastric cancer, acute myeloid leukemia (AML), non-Hodgkin's lymphoma (NHL), ovarian cancer, colorectal cancer, colon cancer, renal cancer, hepatocellular carcinoma, kidney cancer, head and neck cancer, hypopharyngeal squamous cell carcinoma, glioblastoma, neuroblastoma, breast cancer, endometrial cancer, and urothelial cell carcinoma. Although the role of B7-H3 in cancer cells is unclear, its expression may orchestrate signaling events that may protect cancer cells from innate and adaptive immune responses. For example, B7-H3 is overexpressed in high-grade prostatic intraepithelial neoplasia and adenocarcinomas of the prostate, and high expression levels of B7-H3 in these cancerous cells is associated with an increased risk of cancer progression after surgery (Roth et al. Cancer Res. 67(16): 7893-900 (2007)). Further, tumor B7-H3 expression in NSCLC inversely correlated with the number of tumor-infiltrating lymphocytes and significantly correlated with lymph node metastasis (Sun et al. Lung Cancer 53(2): 143-51 (2006)). The level of circulating soluble B7-H3 in NSCLC patients has also been associated with higher tumor stage, tumor size, lymph node metastasis, and distant metastasis (Yamato et al., Br. J. Cancer 101(10):1709-16 (2009)).

B7-H3 may also play an important role in T-cell-mediated antitumor responses in a context dependent manner. For example, gastric cancer tumor cell expression of B7-H3 positively correlated with survival time, infiltration depth, and tissue type (Wu et al., World J. Gastroenterol. 12(3): 457-9 (2006)). Further, high expression of B7-H3 in pancreatic tumor cells was associated with patient survival after surgical resection and significantly correlated with the number of tumor-infiltrating CD8 + T-cells (Loos et al., BMC Cancer 9:463 (2009).

Antibody drug conjugates (ADC) represent a relatively new class of therapeutics comprising an antibody conjugated to a cytotoxic drug via a chemical linker. The therapeutic concept of ADCs is to combine binding capabilities of an antibody with a drug, where the antibody is used to deliver the drug to a tumor cell by means of binding to a target surface antigen, including target surface antigens that are overexpressed in the tumor cells.

There remains a need in the art for anti-B7-H3 antibodies and anti-B7-H3 ADCs that can be used for therapeutic purposes in the treatment of cancer.

›SUMMARY OF THE INVENTION · 1 of 8

In certain aspects, the present invention provides for antibodies and antibody drug conjugates (ADCs) that specifically bind to human B7-H3. In certain aspects, the present invention provides novel ADCs that can selectively deliver Bcl-xL inhibitors to target cancer cells, e.g., B7-H3 expressing cells.

In one aspect, the present invention provides an antoi-B7-H3 antibody, or antigen binding portion thereof, that binds to human B7-H3 (hB7-H3), wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 12 and a light chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 15.

In one embodiment, the anti-B7-H3 antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR2 having the amino acid sequence of SEQ ID NO: 140 and a light chain variable region comprising a CDR2 having the amino acid sequence of SEQ ID NO: 7.

In one embodiment, the anti-B7-H3 antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 10 and a light chain variable region comprising a CDR1 having the amino acid sequence of either SEQ ID NO: 136 or 138.

In one aspect, the present invention provides an anti-B7-H3 antibody, or antigen binding portion thereof, that binds to human B7-H3 (hB7-H3), wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 35 and a light chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 39.

In one embodiment, the anti-B7-H3 antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR2 having the amino acid sequence of SEQ ID NO: 34, and a light chain variable region comprising a CDR2 having the amino acid sequence of SEQ ID NO: 38.

In one embodiment, the anti-B7-H3 antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 33 and a light chain variable region comprising a CDR1 having the amino acid sequence of either SEQ ID NO: 37.

In one embodiment, the anti-B7-H3 antibody, or antigen binding portion thereof, is an IgG isotype.

In one embodiment, the anti-B7-H3 antibody, or antigen binding portion thereof, is an IgG1 or an IgG4 isotype.

In one embodiment, the anti-B7-H3 antibody, or antigen binding portion thereof, has a K D of 1.5×10 −8 or less as determined by surface plasmon resonance.

In one aspect, the present invention provides an anti-B7-H3 antibody, or antigen-binding portion thereof, that binds to hB7-H3, said antibody, or antigen-binding portion thereof, comprising either (i) a heavy chain variable region comprising a CDR set of SEQ ID NOs: 10, 11, and 12, and a light chain variable region comprising a CDR set of SEQ ID NOs: 14, 7, and 15, or (ii) a heavy chain variable region comprising a CDR set of SEQ ID NOs: 33, 34, and 35, and a light chain variable region comprising a CDR set of SEQ ID NOs: 37, 38, and 39.

In one aspect, the present invention provides an anti-B7-H3 antibody that binds to human B7-H3 (hB7-H3), wherein the antibody comprises a heavy chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 12 and a light chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 15.

In one embodiment, the anti-B7-H3 antibody comprises a heavy chain variable region comprising a CDR2 having the amino acid sequence of SEQ ID NO: 140 and a light chain variable region comprising a CDR2 having the amino acid sequence of SEQ ID NO: 7.

In one embodiment, the anti-B7-H3 antibody comprises a heavy chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 10 and a light chain variable region comprising a CDR1 having the amino acid sequence of either SEQ ID NO: 136 or 138.

In one aspect, the present invention provides an anti-B7-H3 antibody that binds to human B7-H3 (hB7-H3), wherein the antibody, or antigen binding portion thereof, comprises a heavy chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 35 and a light chain variable region comprising a CDR3 having the amino acid sequence of SEQ ID NO: 39.

In one embodiment, the anti-B7-H3 antibody comprises a heavy chain variable region comprising a CDR2 having the amino acid sequence of SEQ ID NO: 34, and a light chain variable region comprising a CDR2 having the amino acid sequence of SEQ ID NO: 38.

In one embodiment, the anti-B7-H3 antibody comprises a heavy chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 33 and a light chain variable region comprising a CDR1 having the amino acid sequence of either SEQ ID NO: 37.

In one embodiment, the anti-B7-H3 antibody is an IgG isotype.

In one embodiment, the anti-B7-H3 antibody is an IgG1 or an IgG4 isotype.

In one embodiment, the anti-B7-H3 antibody has a K D of 1.5×10 −8 or less as determined by surface plasmon resonance.

In one aspect, the present invention provides an anti-B7-H3 antibody that binds to hB7-H3, said antibody comprising either (i) a heavy chain variable region comprising a CDR set of SEQ ID NOs: 10, 11, and 12, and a light chain variable region comprising a CDR set of SEQ ID NOs: 14, 7, and 15, or (ii) a heavy chain variable region comprising a CDR set of SEQ ID NOs: 33, 34, and 35, and a light chain variable region comprising a CDR set of SEQ ID NOs: 37, 38, and 39.

In one embodiment, the anti-B7-H3 antibody, or antigen binding portion thereof, is humanized.

In one embodiment, the anti-B7-H3 antibody, or antigen binding portion thereof, further comprises a human acceptor framework. In one embodiment, the human acceptor framework comprises an amino acid sequence selected from the group consisting of SEQ ID Nos: 155, 156, 164, 165, 166, and 167. In one embodiment, the human acceptor framework comprises at least one framework region amino acid substitution. In one embodiment, the amino acid sequence of the framework is at least 65% identical to the sequence of said human acceptor framework and comprises at least 70 amino acid residues identical to said human acceptor framework.

›SUMMARY OF THE INVENTION · 2 of 8

In one embodiment, the human acceptor framework comprises at least one framework region amino acid substitution at a key residue, said key residue selected from the group consisting of a residue adjacent to a CDR; a glycosylation site residue; a rare residue; a residue capable of interacting with human B7-H3; a residue capable of interacting with a CDR; a canonical residue; a contact residue between heavy chain variable region and light chain variable region; a residue within a Vernier zone; and a residue in a region that overlaps between a Chothia-defined variable heavy chain CDR1 and a Kabat-defined first heavy chain framework. In one embodiment, the key residue is selected from the group consisting of 48H, 67H, 69H, 71H, 73H, 94H, and 2L. In one embodiment, the key residue substitution is in the variable heavy chain region and is selected from the group consisting of M48I, V67A, I69L, A71V, K73R, and R94G. In one embodiment, the key residue substitution is in the variable light chain region and is I2V.

In one aspect, the present invention provides an anti-B7-H3 antibody, or antigen-binding portion thereof, that binds to hB7-H3 comprising a heavy chain variable region comprising a CDR set of SEQ ID NOs: 25, 26, and 27, and a light chain variable region comprising a CDR set of SEQ ID NOs: 29, 30, and 31. In one embodiment, the anti-B7-H3 antibody, or antigen binding portion thereof, is humanized In one embodiment, the anti-B7-H3 antibody, or antigen binding portion thereof, further comprises a human acceptor framework.

In one embodiment, the human acceptor framework comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 155 to 158. In one embodiment, the human acceptor framework comprises at least one framework region amino acid substitution. In one embodiment, the amino acid sequence of the framework is at least 65% identical to the sequence of said human acceptor framework and comprises at least 70 amino acid residues identical to said human acceptor framework. In one embodiment, the amino acid sequence of the framework is at least 85% identical, 90% identical, 95% identical, 96% identical, 97% identical, 98% identical, or 99% identical to the sequence of the human acceptor framework and comprises at least 70, at least 75, at least 80, or at least 85 amino acid residues identical to the human acceptor framework.

In one embodiment, the human acceptor framework comprises at least one framework region amino acid substitution at a key residue, said key residue selected from the group consisting of: a residue adjacent to a CDR; a glycosylation site residue; a rare residue; a residue capable of interacting with human B7-H3; a residue capable of interacting with a CDR; a canonical residue; a contact residue between heavy chain variable region and light chain variable region; a residue within a Vernier zone; and a residue in a region that overlaps between a Chothia-defined variable heavy chain CDR1 and a Kabat-defined first heavy chain framework. In one embodiment, the key residue is selected from the group consisting of 69H, 46L, 47L, 64L, and 71L. In one embodiment, the key residue substitution is in the variable heavy chain region and is L69I. In one embodiment, the key residue substitution is in the variable light chain region and is selected from the group consisting of L46P, L47W, G64V, and F71H.

In one aspect, the present invention provides an anti-hB7-H3 antibody, or antigen-binding portion thereof, comprising a heavy chain CDR1 comprising an amino acid sequence as set forth in SEQ ID NO: 10, a heavy chain CDR2 comprising an amino acid sequence as set forth in SEQ ID NO: 140, a heavy chain CDR3 comprising an amino acid sequence as set forth in SEQ ID NO: 12, a light chain CDR1 comprising an amino acid sequence as set forth in SEQ ID NO: 136 or 138, a light chain CDR2 comprising an amino acid sequence as set forth in SEQ ID NO: 7, and a light chain CDR3 comprising an amino acid sequence as set forth in SEQ ID NO: 15.

In another aspect, the present invention provides an anti-hB7-H3 antibody, or antigen-binding portion thereof, comprising a heavy chain CDR1 comprising an amino acid sequence as set forth in SEQ ID NO: 33, a heavy chain CDR2 comprising an amino acid sequence as set forth in SEQ ID NO: 34, a heavy chain CDR3 comprising an amino acid sequence as set forth in SEQ ID NO: 35, a light chain CDR1 comprising an amino acid sequence as set forth in SEQ ID NO: 37, a light chain CDR2 comprising an amino acid sequence as set forth in SEQ ID NO: 38, and a light chain CDR3 comprising an amino acid sequence as set forth in SEQ ID NO: 39.

In one embodiment, the anti-hB7-H3 antibody, or antigen-binding portion thereof, comprises a heavy chain variable domain comprising an amino acid sequence set forth in SEQ ID NO: 139 and a light chain variable domain comprising an amino acid sequence set forth in SEQ ID NO: 135.

In one embodiment, the anti-hB7-H3 antibody, or antigen-binding portion thereof, comprises a heavy chain comprising an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 139, and/or a light chain comprising an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 135.

In one embodiment, the anti-hB7-H3 antibody, or antigen-binding portion thereof, comprises a heavy chain variable domain comprising an amino acid sequence set forth in SEQ ID NO: 147 and a light chain variable domain comprising an amino acid sequence set forth in SEQ ID NO: 144.

›SUMMARY OF THE INVENTION · 3 of 8

In one embodiment, the anti-hB7-H3 antibody, or antigen-binding portion thereof, comprises a heavy chain comprising an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 147, and/or a light chain comprising an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 144.

In one aspect, the present invention provides an anti-hB7-H3 antibody comprising a heavy chain CDR1 comprising an amino acid sequence as set forth in SEQ ID NO: 10, a heavy chain CDR2 comprising an amino acid sequence as set forth in SEQ ID NO: 140, a heavy chain CDR3 comprising an amino acid sequence as set forth in SEQ ID NO: 12, a light chain CDR1 comprising an amino acid sequence as set forth in SEQ ID NO: 136 or 138, a light chain CDR2 comprising an amino acid sequence as set forth in SEQ ID NO: 7, and a light chain CDR3 comprising an amino acid sequence as set forth in SEQ ID NO: 15.

In another aspect, the present invention provides an anti-hB7-H3 antibody comprising a heavy chain CDR1 comprising an amino acid sequence as set forth in SEQ ID NO: 33, a heavy chain CDR2 comprising an amino acid sequence as set forth in SEQ ID NO: 34, a heavy chain CDR3 comprising an amino acid sequence as set forth in SEQ ID NO: 35, a light chain CDR1 comprising an amino acid sequence as set forth in SEQ ID NO: 37, a light chain CDR2 comprising an amino acid sequence as set forth in SEQ ID NO: 38, and a light chain CDR3 comprising an amino acid sequence as set forth in SEQ ID NO: 39.

In one embodiment, the anti-hB7-H3 antibody comprises a heavy chain variable domain comprising an amino acid sequence set forth in SEQ ID NO: 139 and a light chain variable domain comprising an amino acid sequence set forth in SEQ ID NO: 135.

In one embodiment, the anti-hB7-H3 antibody comprises a heavy chain comprising an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 139, and/or a light chain comprising an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 135.

In one embodiment, the anti-hB7-H3 antibody comprises a heavy chain variable domain comprising an amino acid sequence set forth in SEQ ID NO: 147 and a light chain variable domain comprising an amino acid sequence set forth in SEQ ID NO: 144.

In one embodiment, the anti-hB7-H3 antibody comprises a heavy chain comprising an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 147, and/or a light chain comprising an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 144.

In one embodiment, the anti-hB7-H3 antibody, or antigen-binding portion thereof, comprises a heavy chain CDR set corresponding to antibody huAb13v1, and a light chain CDR set corresponding to antibody huAb13v1. In one embodiment, the anti-hB7-H3 antibody, or antigen-binding portion thereof, comprises a heavy chain variable region corresponding to antibody huAb13v1, and a light chain variable region corresponding to antibody huAb13v1.

In one embodiment, the anti-hB7-H3 antibody, or antigen-binding portion thereof, comprises a heavy chain CDR set corresponding to antibody huAb3v2.5, and a light chain CDR set corresponding to antibody huAb3v2.5. In one embodiment, the anti-hB7-H3 antibody, or antigen-binding portion thereof, comprises a heavy chain variable region corresponding to antibody huAb3v2.5, and a light chain variable region corresponding to antibody huAb3v2.5.

In one embodiment, the antibody, or antigen binding portion thereof, binds cynomolgus B7-H3.

In one embodiment, the antibody, or antigen binding portion thereof, has a dissociation constant (K D ) to hB7-H3 selected from the group consisting of: at most about 10 −7 M; at most about 10 −8 M; at most about 10 −9 M; at most about 10 −1 ° M; at most about 10 −11 M; at most about 10 −12 M; and at most 10 −13 M.

In one embodiment, the antibody, or antigen binding portion thereof, comprises a heavy chain immunoglobulin constant domain of a human IgM constant domain, a human IgG1 constant domain, a human IgG2 constant domain, a human IgG3 constant domain, a human IgG4 constant domain, a human IgA constant domain, or a human IgE constant domain.

In one embodiment, the antibody is an IgG having four polypeptide chains which are two heavy chains and two light chains.

In one embodiment, the antibody, or antigen-binding portion thereof, comprises the heavy chain immunoglobulin constant region domain is a human IgG1 constant domain. In one embodiment, the human IgG1 constant domain comprises an amino acid sequence of SEQ ID NO: 159 or SEQ ID NO: 160.

In one aspect, the present invention provides an isolated antibody, or antigen binding portion thereof, that binds to human B7-H3 (hB7-H3), wherein the antibody, or antigen binding portion thereof, comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 168 and a light chain comprising the amino acid sequence of SEQ ID NO: 169.

In one aspect, the present invention provides an isolated antibody, or antigen binding portion thereof, that binds to human B7-H3 (hB7-H3), wherein the antibody, or antigen binding portion thereof, comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 170 and a light chain comprising the amino acid sequence of SEQ ID NO: 171.

In one aspect, the present invention provides an isolated antibody, or antigen binding portion thereof, that binds to human B7-H3 (hB7-H3), wherein the antibody, or antigen binding portion thereof, comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 172 and a light chain comprising the amino acid sequence of SEQ ID NO: 173.

›SUMMARY OF THE INVENTION · 4 of 8

In one embodiment, the antibody, or antigen binding portion thereof, further comprises a light chain immunoglobulin constant domain comprising a human Ig kappa constant domain or a human Ig lambda constant domain.

In one embodiment, the anti-hB7-H3 antibody, or antigen-binding portion thereof, competes with the antibody, or antigen binding portion thereof, of any one of the anti-hB7-H3 antibodies, or antigen-binding portions thereof, disclosed herein.

In one aspect, the present invention provides a pharmaceutical composition comprising the anti-hB7-H3 antibody, or antigen binding portion thereof, as disclosed herein, and a pharmaceutically acceptable carrier.

In another aspect, the present invention provides an anti-hB7-H3 Antibody Drug Conjugate (ADC) comprising an anti-hB7-H3 antibody disclosed herein conjugated to a drug via a linker In one embodiment, the drug is an auristatin or a pyrrolobenzodiazepine (PBD). In one embodiment, the drug is a Bcl-xL inhibitor.

In one aspect, the present invention provides an anti-hB7-H3 antibody drug conjugate (ADC) comprising a drug linked to an anti-human B7-H3 (hB7-H3) antibody by way of a linker, wherein the drug is a Bcl-xL inhibitor according to structural formula (IIa), (IIb), (IIc), or (IId):

wherein:

Ar 1 is selected from

and is optionally substituted with one or more substituents independently selected from halo, hydroxy, nitro, lower alkyl, lower heteroalkyl, C 1-4 alkoxy, amino, cyano and halomethyl;

Ar 2 is selected from

or an N-oxide thereof, and is optionally substituted with one or more substituents independently selected from halo, hydroxy, nitro, lower alkyl, lower heteroalkyl, C 1-4 alkoxy, amino, cyano and halomethyl, wherein the R 12 —Z 2b —, R′—Z 2b —, #—N(R 4 )—R 13 —Z 2b —, or #—R′—Z 2b — substituents are attached to Ar 2 at any Ar 2 atom capable of being substituted; Z′ is selected from N, CH, C-halo, C—CH 3 and C—CN; Z 2a and Z 2b are each, independently from one another, selected from a bond, NR 6 , CR 6a R 6b , O, S, S(O), S(O) 2 , NR 6 C(O)—, —NR 6a C(O)NR 6b —, and NR 6 C(O)O—;

R′ is

wherein #, where attached to R′, is attached to R′ at any R′ atom capable of being substituted; X′ is selected at each occurrence from —N(R 10 )—, —N(R 10 )C(O)—, —N(R 10 )S(O) 2 —, —S(O) 2 N(R 10 )—, and —O—; n is selected from 0-3; R 10 is independently selected at each occurrence from hydrogen, lower alkyl, heterocycle, aminoalkyl, G-alkyl, and —(CH 2 ) 2 —O—(CH 2 ) 2 —O—(CH 2 ) 2 —NH 2 ; G at each occurrence is independently selected from a polyol, a polyethylene glycol with between 4 and 30 repeating units, a salt and a moiety that is charged at physiological pH; SP a is independently selected at each occurrence from oxygen, —S(O) 2 N(H)—, —N(H)S(O) 2 —, —N(H)C(O)—, —C(O)N(H)—, —N(H)—, arylene, heterocyclene, and optionally substituted methylene; wherein methylene is optionally substituted with one or more of —NH(CH 2 ) 2 G, NH 2 , C 1-8 alkyl, and carbonyl; m 2 is selected from 0-12; R 1 is selected from hydrogen, methyl, halo, halomethyl, ethyl, and cyano; R 2 is selected from hydrogen, methyl, halo, halomethyl and cyano; R 3 is selected from hydrogen, methyl, ethyl, halomethyl and haloethyl; R 4 is selected from hydrogen, lower alkyl and lower heteroalkyl or is taken together with an atom of R 13 to form a cycloalkyl or heterocyclyl ring having between 3 and 7 ring atoms; R 6 , R 6a and R 6b are each, independent from one another, selected from hydrogen, optionally substituted lower alkyl, optionally substituted lower heteroalkyl, optionally substituted cycloalkyl and optionally substituted heterocyclyl, or are taken together with an atom from R 4 and an atom from R 13 to form a cycloalkyl or heterocyclyl ring having between 3 and 7 ring atoms; R 11a and R 11b are each, independently of one another, selected from hydrogen, halo, methyl, ethyl, halomethyl, hydroxyl, methoxy, CN, and SCH 3 ; R 12 is optionally R′ or is selected from hydrogen, halo, cyano, optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, and optionally substituted cycloalkyl; R 13 is selected from optionally substituted C 1-8 alkylene, optionally substituted heteroalkylene, optionally substituted heterocyclene, and optionally substituted cycloalkylene; and # represents a point of attachment to a linker; and wherein the anti-hB7-H3 antibody binds to B7-H3 (SEQ ID NO: 149) with a dissociation constant (K d ) of about 1×10 −6 M or less. In a further embodiment, the antibody comprises a heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO: 147 and a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO: 144; binds to B7-H3 (SEQ ID NO: 149) with a dissociation constant (K d ) of about 1×10 −6 M or less, as determined by surface plasmon resonance; and/or inhibits tumor growth in an in vivo human small-cell lung carcinoma (SCLC) xenograft assay with a tumor growth inhibition % (TGI %) of at least about 50% relative to a human IgG antibody which is not specific for B7-H3, wherein the human IgG antibody is administered in the SCLC xenograft assay at the same dose and frequency as the anti-hB7-H3 antibody.

In one embodiment, the ADC is a compound according to structural formula (I):

(D-L-LK m Ab (I)

wherein D is the Bcl-xL inhibitor drug of formula (IIa), (IIb), (IIc) or (IId); L is the linker; Ab is the anti-hB7-H3 antibody; LK represents a covalent linkage linking the linker (L) to the anti-hB7-H3 antibody (Ab); and m is an integer ranging from 1 to 20.

In one embodiment, G at each occurrence is a salt or a moiety that is charged at physiological pH.

In one embodiment, G at each occurrence is a salt of a carboxylate, a sulfonate, a phosphonate, or ammonium.

In one embodiment, G at each occurrence is a moiety that is charged at physiological pH selected from the group consisting of carboxylate, a sulfonate, a phosphonate, and an amine.

In one embodiment, G at each occurrence is a moiety containing a polyethylene glycol with between 4 and 30 repeating units, or a polyol.

›SUMMARY OF THE INVENTION · 5 of 8

In one embodiment, the polyol is a sugar.

In one embodiment, the ADC is of the formula (IIa) or formula (IId), and R′ includes at least one substitutable nitrogen suitable for attachment to a linker.

In one embodiment, G is selected at each occurrence from:

wherein M is hydrogen or a positively charged counterion.

In one embodiment, R′ is selected from

wherein # represents either a hydrogen atom in the Bcl-xL inhibitor drug of the ADCs of formula (IIb) or (IIc) or the point of attachment in the Bcl-xL inhibitor drug of the ADCs of formula (IIa) or (IId) to a linker L.

In one embodiment, Ar 1 is selected from

and is optionally substituted with one or more substituents independently selected from halo, cyano, methyl, and halomethyl.

In one embodiment, Ar 1 is

In one embodiment, Ar 2 is

optionally substituted with one or more substituents.

In one embodiment, Ar 2 is selected from

and is optionally substituted with one or more substituents.

In one embodiment, Ar 2 is substituted with one or more solubilizing groups.

In one embodiment, each solubilizing group is, independently of the others, selected from a moiety containing a polyol, a polyethylene glycol with between 4 and 30 repeating units, a salt, or a moiety that is charged at physiological pH.

In one embodiment, Ar 2 is substituted with one or more solubilizing groups.

In one embodiment, Z 1 is N.

In one embodiment, Z 2a is O.

In one embodiment, R 1 is methyl or chloro.

In one embodiment, R 2 is hydrogen or methyl.

In one embodiment, R 2 is hydrogen.

In one embodiment, Z 2b is O.

In one embodiment, Z 2b is NH or CH 2 .

In one embodiment, the ADC is a compound according to structural formula (IIa).

In one embodiment, the ADC is a compound according to structural formula (IIa) which includes a core selected from structures (C.1)-(C.21):

In one embodiment, the ADC is a compound according to structural formula (IIa.1):

wherein Y is optionally substituted C 1 -C 8 alkylene; r is 0 or 1; and s is 1, 2 or 3.

In one embodiment, the ADC is a compound according to structural formula (IIa.2):

wherein U is selected from N, O and CH, with the proviso that when U is O, then V a and R 21a are absent; R 20 is selected from H and C 1 -C 4 alkyl; R 21a and R 21b are each, independently from one another, absent or selected from H, C 1 -C 4 alkyl and G, where G is selected from a polyol, PEG4-30, a salt and a moiety that is charged at physiological pH; V a and V b are each, independently from one another, absent or selected from a bond, and an optionally substituted alkylene; R 20 is selected from H and C 1 -C 4 alkyl; and s is 1, 2 or 3.

In one embodiment, the ADC is a compound according to structural formula (IIa.3):

wherein R b is selected from H, C 1 -C 4 alkyl and J b -G or is optionally taken together with an atom of T to form a ring having between 3 and 7 atoms; J a and J b are each, independently from one another, selected from optionally substituted C 1 -C 8 alkylene and optionally substituted phenylene; T is selected from optionally substituted C 1 -C 5 alkylene, CH 2 CH 2 OCH 2 CH 2 OCH 2 CH 2 , CH 2 CH 2 OCH 2 CH 2 OCH 2 CH 2 OCH 2 and a polyethylene glycol containing from 4 to 10 ethylene glycol units; G is selected from a polyol, PEG4-30, a salt and a moiety that is charged at physiological pH; and s is 1, 2 or 3.

In one embodiment, the ADC is a compound according to structural formula (IIb).

In one embodiment, the ADC is a compound according to structural formula (IIb.1):

wherein Y is optionally substituted C 1 -C 8 alkylene; G is selected from a polyol, PEG4-30, a salt and a moiety that is charged at physiological pH; r is 0 or 1; and s is 1, 2 or 3.

In one embodiment, the ADC is a compound according to structural formula (IIc).

In one embodiment, the ADC is a compound according to structural formula (IIc.1):

wherein Y a is optionally substituted C 1 -C 8 alkylene; Y b is optionally substituted C 1 -C 8 alkylene; R 23 is selected from H and C 1 -C 4 alkyl; and G is selected from a polyol, PEG4-30, a salt and a moiety that is charged at physiological pH.

In one embodiment, the ADC is a compound according to structural formula (IIc.2):

wherein Y a is optionally substituted C 1 -C 8 alkylene; Y b is optionally substituted C 1 -C 8 alkylene; Y c is optionally substituted C 1 -C 8 alkylene; R 23 is selected from H and C 1 -C 4 alkyl; R 25 is Y b -G or is taken together with an atom of Y c to form a ring having 4-6 ring atoms; and G is selected from a polyol, PEG4-30, a salt and a moiety that is charged at physiological pH.

In one embodiment, the Bcl-xL inhibitor is selected from the group consisting of the following compounds modified in that the hydrogen corresponding to the # position of structural formula (IIa), (IIb), (IIc), or (IId) is not present forming a monoradical:

6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-[1-({3-[2-({2-[2-(carboxymethoxy)ethoxy]ethyl}amino)ethoxy]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 2-{[(2-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]amino}ethyl)sulfonyl]amino}-2-deoxy-D-glucopyranose; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(4-{[(3R,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl]methyl}benzyl)amino]ethoxy}tricyclo[3.3.1.13,7]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(3-sulfopropyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[(2,3-dihydroxyprop yl)amino]ethoxy]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 2-({[4-({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]amino}methyl)phenyl]sulfonyl}amino)-2-deoxy-beta-D-glucopyranose; 8-(1,3-benzothiazol-2-ylcarbamoyl)-2-{6-carboxy-5-[1-({3-[2-({2-[1-(beta-D-glucopyranuronosyl)-1H-1,2,3-triazol-4-yl]ethyl}amino)ethoxy]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridin-2-yl}-1,2,3,4-tetrahydroisoquinoline; 3-[1-({3-[2-(2-[{4-(beta-D-allopyranosyloxy)benzyl]amino}ethoxy)ethoxy]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3,5-dimethyl-7-(2-{2-[(2-sulfoethyl)amino]ethoxy}ethoxy)tricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-phosphonoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[methyl(3-sulfo-L-alanyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(3-phosphonopropyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(3-sulfo-L-alanyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3,5-dimethyl-7-(2-{2-[(3-phosphonopropyl)amino]ethoxy}ethoxy)tricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; 3-{1-[(3-{2-[L-alpha-aspartyl(methyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carb oxylic acid; 6-{4-[({2-[2-(2-aminoethoxy)ethoxy]ethyl}[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]amino)methyl]benzyl}-2,6-anhydro-L-acid; 4-({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]amino}methyl)phenyl hexopyranosiduronic acid; 6-[1-(1,3-benzothiazol-2-ylcarbamoyl)-1,2,3,4-tetrahydroquinolin-7-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-phosphonoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[1-(1,3-benzothiazol-2-ylcarbamoyl)-1,2,3,4-tetrahydroquinolin-7-yl]-3-{1-[(3,5-dimethyl-7-{2-[methyl(3-sulfo-L-alanyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-([1,3]thiazolo[5,4-b]pyridin-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-([1,3]thiazolo[4,5-b]pyridin-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 6-[1-(1,3-benzothiazol-2-ylcarbamoyl)-1,2,3,4-tetrahydroquinolin-7-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[(2-carboxyethyl)amino]ethoxy]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(3-phosphonopropyl)(piperidin-4-yl)amino}ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 3-{1-[(3-{2-[D-alpha-aspartyl(methyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carb oxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3-(2-{[1-(carboxymethyl)piperidin-4-yl]amino}ethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; N-[(5S)-5-amino-6-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](methyl)amino}-6-oxohexyl]-N,N-dimethylmethanaminium; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[piperidin-4-yl(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-5-(3-phosphonopropoxy)-3,4-dihydroisoquinolin-2(1H)-yl]-3-[1-({3,5-dimethyl-7-[2-(methylamino)ethoxy]tricyclo[3.3.1.1 3,7 ]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3-(2-{[N-(2-carboxyethyl)-L-alpha-aspartyl]amino}ethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; 3-{1-[(3-{2-[(2-aminoethyl)(2-sulfoethyl))amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 6-[5-(2-aminoethoxy)-8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-[1-({3,5-dimethyl-7-[2-(methylamino)ethoxy]tricyclo[3.3.1.1 3,7 ]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-3-{1-[(3,5-dimethyl-7-{2-[(3-sulfopropyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[(2-carboxyethyl)(piperidin-4-yl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(3-sulfo-L-alanyl)(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[{2-[(2-carboxyethyl)amino]ethyl}(2-sulfoethyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 3-{1-[(3,5-dimethyl-7-{2-[(3-phosphonopropyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-([1,3]thiazolo[4,5-b]pyridin-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 3-{1-[(3,5-dimethyl-7-{2-[(3-phosphonopropyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-([1,3]thiazolo[5,4-b]pyridin-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-5-(carboxymethoxy)-3,4-dihydroisoquinolin-2(1H)-yl]-3-[1-({3,5-dimethyl-7-[2-(methylamino)ethoxy]tricyclo[3.3.1.1 3,7 ]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[(3-carboxypropyl)(piperidin-4-yl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 3-{1-[(3-{2-[L-alpha-aspartyl(2-sulfoethyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[(1,3-dihydroxypropan-2-yl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[5-(2-aminoethoxy)-8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[methyl(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-5-{2-[(2-sulfoethyl)amino]ethoxy}-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[methyl(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl) {2-[(2-sulfoethyl)amino]ethyl}amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-5-{2-[(2-carboxyethyl)amino]ethoxy}-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[methyl(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 3-{1-[(3,5-dimethyl-7-{2-[(3-phosphonopropyl)(piperidin-4-yl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-([1,3]thiazolo[4,5-b]pyridin-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 6-[4-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydro-2H-1,4-benzoxazin-6-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-5-(3-sulfopropoxy)-3,4-dihydroisoquinolin-2(1H)-yl]-3-[1-({3,5-dimethyl-7-[2-(methylamino)ethoxy]tricyclo[3.3.1.1 3,7 ]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridine-2-carboxylic acid; 3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[1-([1,3]thiazolo[4,5-b]pyridin-2-ylcarbamoyl)-1,2,3,4-tetrahydroquinolin-7-yl]pyridine-2-carboxylic acid; 3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-([1,3]thiazolo[4,5-b]pyridin-2-ylcarbamoyl)naphthalen-2-yl]pyridine-2-carboxylic acid; (1ξ)-1-({2-[5-(1-{[3-(2-aminoethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)-6-carboxypyridin-2-yl]-8-(1,3-benzothiazol-2-ylcarbamoyl)-1,2,3,4-tetrahydroisoquinolin-5-yl}methyl)-1,5-anhydro-D-glucitol; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[(3-carboxypropyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-3-{1-[(3,5-dimethyl-7-{2-[(3-phosphonopropyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3-(2-{[4-(beta-D-glucopyranosyloxy)benzyl]amino}ethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; 3-(1-{[3-(2-{[4-(beta-D-allopyranosyloxy)benzyl]amino}ethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 3-{1-[(3-{2-[azetidin-3-yl(2-sulfoethyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 3-{1-[(3-{2-[(3-aminopropyl)(2-sulfoethyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 6-[1-(1,3-benzothiazol-2-ylcarbamoyl)-1,2,3,4-tetrahydroquinolin-7-yl]-3-{1-[(3-{2-[(2-carboxyethyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[(N 6 ,N 6 -dimethyl-L-lysyl)(methyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 3-{1-[(3-{2-[(3-aminopropyl)(methyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[1-(1,3-benzothiazol-2-ylcarbamoyl)-1,2,3,4-tetrahydroquinolin-7-yl]pyridine-2-carboxylic acid; 3-{1-[(3-{2-[azetidin-3-yl(methyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[1-(1,3-benzothiazol-2-ylcarbamoyl)-1,2,3,4-tetrahydroquinolin-7-yl]pyridine-2-carboxylic acid; N 6 -(37-oxo-2,5,8,11,14,17,20,23,26,29,32,35-dodecaoxaheptatriacontan-37-yl)-L-lysyl-N-[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]-L-alaninamide; methyl 6-[4-(3-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]amino}propyl)-1H-1,2,3-triazol-1-yl]-6-deoxy-beta-L-glucopyranoside; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-3-{1-[(3-{2-[(2-carboxyethyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[5-(1,3-benzothiazol-2-ylcarbamoyl)quinolin-3-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[4-(1,3-benzothiazol-2-ylcarbamoyl)quinolin-6-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[5-(1,3-benzothiazol-2-ylcarbamoyl)quinolin-3-yl]-3-{1-[(3-{2-[(2-carboxyethyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[1-(1,3-benzothiazol-2-ylcarbamoyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 8-(1,3-benzothiazol-2-ylcarbamoyl)-2-{6-carboxy-5-[1-({3-[2-({3-[1-(beta-D-glucopyranuronosyl)-1H-1,2,3-triazol-4-yl]propyl}amino)ethoxy]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridin-2-yl}-1,2,3,4-tetrahydroisoquinoline; 6-[7-(1,3-benzothiazol-2-ylcarbamoyl)-1H-indol-2-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-6-[3-(methylamino)propyl]-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 5-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]amino}-5-deoxy-D-arabinitol; 1-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]amino}-1,2-dideoxy-D-arabino-hexitol; 6-[4-(1,3-benzothiazol-2-ylcarbamoyl)isoquinolin-6-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3-(2-{[3-hydroxy-2-(hydroxymethyl)propyl]amino}ethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; 1-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]amino}-1,2-dideoxy-D-erythro-pentitol; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3,5-dimethyl-7-(2-{[(2S,3S)-2,3,4-trihydroxybutyl]amino}ethoxy)tricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3-(2-{[(2S,3S,4R,5R,6R)-2,3,4,5,6,7-hexahydroxyheptyl]amino}ethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[({3-[(1,3-dihydroxypropan-2-yl)amino]propyl}sulfonyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[(3-{[1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl]amino}-3-oxopropyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3-(2-{[(3S)-3,4-dihydroxybutyl]amino}ethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; 4-({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]amino}methyl)phenyl beta-D-glucopyranosiduronic acid; 3-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]amino}propyl beta-D-glucopyranosiduronic acid; 6-[4-(1,3-benzothiazol-2-ylcarbamoyl)-2-oxidoisoquinolin-6-yl]-3-[1-({3,5-dimethyl-7-[2-(methylamino)ethoxy]tricyclo[3.3.1.1 3,7 ]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridine-2-carboxylic acid; 6-{8-[(1,3-benzothiazol-2-yl)carbamoyl]-3,4-dihydroisoquinolin-2(1H)-yl}-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]acetamido}tricyclo[3.3.1.1 3,7 ]decan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3,5-dimethyl-7-({2-[(2-sulfoethyl)amino]ethyl}sulfanyl)tricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; and 6-{8-[(1,3-benzothiazol-2-yl)carbamoyl]-3,4-dihydroisoquinolin-2(1H)-yl}-3-{1-[(3,5-dimethyl-7-{3-[(2-sulfoethyl)amino]propyl}tricyclo[3.3.1.1 3,7 ]decan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid.

›SUMMARY OF THE INVENTION · 6 of 8

In one embodiment, the linker is cleavable by a lysosomal enzyme. In one embodiment, the lysosomal enzyme is Cathepsin B.

In one embodiment, the linker comprises a segment according to structural formula (IVa), (IVb), (IVc), or (IVd):

wherein peptide represents a peptide (illustrated N→C, wherein peptide includes the amino and carboxy “termini”) a cleavable by a lysosomal enzyme; T represents a polymer comprising one or more ethylene glycol units or an alkylene chain, or combinations thereof;

R a is selected from hydrogen, C 16 alkyl, SO 3 H and CH 2 SO 3 H; R y is hydrogen or C 1-4 alkyl-(O) 1 —(C 1-4 alkylene) s -G 1 or C 1-4 alkyl-(N)—[(C 1-4 alkylene)-G 1 ]2; R z is C 1-4 alkyl-(O), —(C 1-4 alkylene) s -G 2 ; G 1 is SO 3 H, CO 2 H, PEG 4-32, or sugar moiety; G 2 is SO 3 H, CO 2 H, or PEG 4-32 moiety; r is 0 or 1; s is 0 or 1; p is an integer ranging from 0 to 5; q is 0 or 1; x is 0 or 1; y is 0 or 1; represents the point of attachment of the linker to the Bcl-xL inhibitor; and * represents the point of attachment to the remainder of the linker.

In one embodiment, the peptide is selected from the group consisting of Val-Cit; Cit-Val; Ala-Ala; Ala-Cit; Cit-Ala; Asn-Cit; Cit-Asn; Cit-Cit; Val-Glu; Glu-Val; Ser-Cit; Cit-Ser; Lys-Cit; Cit-Lys; Asp-Cit; Cit-Asp; Ala-Val; Val-Ala; Phe-Lys; Lys-Phe; Val-Lys; Lys-Val; Ala-Lys; Lys-Ala; Phe-Cit; Cit-Phe; Leu-Cit; Cit-Leu; Ile-Cit; Cit-Ile; Phe-Arg; Arg-Phe; Cit-Trp; and Trp-Cit.

In one embodiment, the lysosomal enzyme is f-glucuronidase or β-galactosidase.

In one embodiment, the linker comprises a segment according to structural formula (Va), (Vb), (Vc), (Vd), or (Ve):

wherein q is 0 or 1; r is 0 or 1; X 1 is CH 2 , O or NH; represents the point of attachment of the linker to the drug; and * represents the point of attachment to the remainder of the linker.

In one embodiment, the linker comprises a segment according to structural formula (VIIIa), (VIIIb), or (VIIIc):

or a hydrolyzed derivative thereof, wherein: R 9 is H or —O—(CH 2 CH 2 O) 11 —CH 3 ; x is 0 or 1; y is 0 or 1; G 3 is —CH 2 CH 2 CH 2 SO 3 H or —CH 2 CH 2 O—(CH 2 CH 2 O) 11 —CH 3 ; R w is —O—CH 2 CH 2 SO 3 H or —NH(CO)—CH 2 CH 2 O(CH CH 2 O) 12 —CH 3 ; * represents the point of attachment to the remainder of the linker; and represents the point of attachment of the linker to the antibody.

In one embodiment, the linker comprises a polyethylene glycol segment having from 1 to 6 ethylene glycol units.

In one embodiment, m is 2, 3 or 4.

In one embodiment, linker L is selected from IVa or IVb.

In one embodiment, linker L is selected from the group consisting of IVa.1-IVa.8, IVb.1-IVb.19, IVc.1-IVc.7, IVd.1-IVd.4, Va.1-Va.12, Vb.1-Vb.10, Vc. 1-Vc.11, Vd.1-Vd.6, Ve.1-Ve.2, VIa.1, VIc.1-VIc.2, VId.1-VId.4, VIIa.1-VIIa.4, VIIb.1-VIIb.8, VIIc.1-VIIc.6 in either the closed or open form.

In one embodiment, the linker L is selected from the group consisting of IVb.2, IVc.5, IVc.6, IVc.7, IVd.4, Vb.9, VIIa.1, VIIa.3, VIIc.1, VIIc.4, and VIIc.5, wherein the maleimide of each linker has reacted with the antibody Ab, forming a covalent attachment as either a succinimide (closed form) or succinamide (open form).

In one embodiment, the linker L is selected from the group consisting of IVb.2, IVc.5, IVc.6, IVd.4, VIIa.1, VIIa.3, VIIc.1, VIIc.4, VIIc.5, wherein the maleimide of each linker has reacted with the antibody Ab, forming a covalent attachment as either a succinimide (closed form) or succinamide (open form).

In one embodiment, the linker L is selected from the group consisting of IVb.2, VIIa.3, IVc.6, and VIIc.1, wherein is the attachment point to drug D and @ is the attachment point to the LK, wherein when the linker is in the open form as shown below, @ can be either at the α-position or β-position of the carboxylic acid next to it:

In one embodiment, LK is a linkage formed with an amino group on the anti-hB7H3 antibody Ab.

In one embodiment, LK is an amide or a thiourea.

In one embodiment, LK is a linkage formed with a sulfhydryl group on the anti-hB7-H3 antibody Ab.

In one embodiment, LK is a thioether.

In one embodiment, LK is selected from the group consisting of amide, thiourea and thioether; and m is an integer ranging from 1 to 8.

In one embodiment, D is a Bcl-xL inhibitor as described herein; L is selected from the group consisting of linkers IVa.1-IVa.8, IVb.1-IVb.19, IVc.1-IVc.7, IVd.1-IVd.4, Va.1-Va.12, Vb.1-Vb.10, Vc.1-Vc.11, Vd.1-Vd.6, Ve.1-Ve.2, VIa.1, VIc.1-Vlc.2, VId.1-VId.4, VIIa.1-VIIa.4, VIIb.1-VIIb.8, and VIIc.1-VIIc.6, wherein each linker has reacted with the antibody, Ab, forming a covalent attachment; LK is thioether; and m is an integer ranging from 1 to 8.

In one embodiment, D is the Bcl-xL inhibitor selected from the group consisting of the following compounds modified in that the hydrogen corresponding to the # position of structural formula (IIa), (IIb), (IIc), or (IId) is not present, forming a monoradical:

6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[(2-carboxyethyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 1-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]amino}-1,2-dideoxy-D-arabino-hexitol; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3-(2-{[3-hydroxy-2-(hydroxymethyl)propyl]amino}ethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; and 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3-(2-{[(3S)-3,4-dihydroxybutyl]amino}ethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid;

›SUMMARY OF THE INVENTION · 7 of 8

L is selected from the group consisting of linkers IVb.2, IVc.5, IVc.6, IVc.7, IVd.4, Vb.9, Vc.11, VIIa.1, VIIa.3, VIIc.1, VIIc.4, and VIIc.5 in either closed or open forms; LK is thioether; and

m is an integer ranging from 2 to 4.

In one embodiment, the ADC is selected from the group consisting of formulae i-vi:

wherein m is an integer from 1 to 6. In one embodiment, Ab is an anti-hB7-H3 antibody, wherein the anti-hB7-H3 antibody comprises a heavy chain CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 35, a heavy chain CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 34, and a heavy chain CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 33; and a light chain CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 39, a light chain CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 38, and a light chain CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 37. In one embodiment, the Ab is an anti-hB7-H3 antibody, wherein the anti-hB7H3 antibody comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 147, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 144. In one embodiment, Ab is an anti-hB7-H3 antibody, wherein the anti-hB7-H3 antibody comprises a heavy chain constant region comprising the amino acid sequence set forth in SEQ ID NO: 160 and/or a light chain constant region comprising the amino acid sequence set forth in SEQ ID NO: 161. In one embodiment, Ab is an anti-hB7-H3 antibody, wherein the anti-hB7-H3 antibody comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 168, and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 169. In one embodiment, Ab is an anti-hB7-H3 antibody, wherein the anti-hB7-H3 antibody comprises a heavy chain CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 12, a heavy chain CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 140, and a heavy chain CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 10; and a light chain CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 15, a light chain CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a light chain CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 136. In one embodiment, the Ab is an anti-hB7-H3 antibody, wherein the anti-hB7H3 antibody comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 139, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 135. In one embodiment, Ab is an anti-hB7-H3 antibody, wherein the anti-hB7-H3 antibody comprises a heavy chain constant region comprising the amino acid sequence set forth in SEQ ID NO: 160 and/or a light chain constant region comprising the amino acid sequence set forth in SEQ ID NO: 161. In one embodiment, Ab is an anti-hB7-H3 antibody, wherein the anti-hB7-H3 antibody comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 170, and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 171.

In one embodiment, m is an integer from 2 to 6. In one embodiment, m is 2.

In one embodiment, the ADC comprises an anti-hB7-H3 antibody comprising a heavy chain CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 12, a heavy chain CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 140, and a heavy chain CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 10; a light chain CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 15, a light chain CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a light chain CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 136 or 138.

In one embodiment, the ADC comprises an antibody comprising a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 139, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 135.

In one embodiment, the ADC comprises an the antibody comprising a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 139, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 137.

In one embodiment, the ADC comprises an antibody comprising a light chain CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 39, a light chain CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 38, and a light chain CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 37; and a heavy chain CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 35, a heavy chain CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 34, and a heavy chain CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 33.

In one embodiment, the ADC comprises an antibody comprising a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 147, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 144.

In one embodiment, the ADC is selected from the group consisting of huAb3v2.5-CZ, huAb3v2.5-TX, huAb3v2.5-TV, huAb3v2.5-YY, huAb3v2.5-AAA, huAb3v2.5-AAD, huAb3v2.6-CZ, huAb3v2.6-TX, huAb3v2.6-TV, huAb3v2.6-YY, huAb3v2.6-AAD, huAb13v1-CZ, huAb13v1-TX, huAb13v1-TV, huAb13v1-YY, huAb13v1-AAA, huAb13v1-AAD, wherein CZ, TX, TV, YY, AAA, and AAD are synthons disclosed in Table B, and wherein the conjugated synthons are either in open or closed form.

In one aspect, the present invention provides a pharmaceutical composition comprising an effective amount of an ADC described herein, and a pharmaceutically acceptable carrier.

In another aspect, the present invention provides a pharmaceutical composition comprising an ADC mixture comprising a plurality of ADCs described herein, and a pharmaceutically acceptable carrier.

›SUMMARY OF THE INVENTION · 8 of 8

In one embodiment, the ADC mixture has an average drug to antibody ratio (DAR) of 1.5 to 4.

In one embodiment, the ADC mixture comprises ADCs each having a DAR of 1.5 to 8.

In one aspect, the present invention provides a method for treating cancer, comprising administering a therapeutically effective amount of an ADC described herein to a subject in need thereof.

In one embodiment, the cancer is selected from the group consisting of small cell lung cancer, non small cell lung cancer, breast cancer, ovarian cancer, a glioblastoma, prostate cancer, pancreatic cancer, colon cancer, gastric cancer, melanoma, hepatocellular carcinoma, head and neck cancer, kidney cancer, leukemia, e.g., acute myeloid leukemia (AML), and lymphoma, e.g., non-Hodgkin's lymphoma (NHL).

In one embodiment, the cancer is a squamous cell carcinoma. In one embodiment, the squamous cell carcinoma is squamous lung cancer or squamous head and neck cancer.

In one embodiment, the cancer is triple negative breast cancer.

In one embodiment, the cancer is non-small cell lung cancer.

In one embodiment, the cancer is characterized as having an activating EGFR mutation. In one embodiment, the activating EGFR mutation is selected from the group consisting of an exon 19 deletion mutation, a single-point substitution mutation L858R in exon 21, a T790M point mutation, and combinations thereof.

In one aspect, the present invention provides a method for inhibiting or decreasing solid tumor growth in a subject having a solid tumor, said method comprising administering an effective amount of an ADC described herein to the subject having the solid tumor, such that the solid tumor growth is inhibited or decreased.

In one embodiment, the solid tumor is a non-small cell lung carcinoma.

In one embodiment, the ADC is administered in combination with an additional agent or an additional therapy.

In one embodiment, the additional agent is selected from the group consisting of an anti-PD1 antibody (e.g. pembrolizumab), an anti-PD-L1 antibody (e.g., atezolizumab), an anti-CTLA-4 antibody (e.g. ipilimumab), a MEK inhibitor (e.g. trametinib), an ERK inhibitor, a BRAF inhibitor (e.g. dabrafenib), osimertinib, erlotinib, gefitinib, sorafenib, a CDK9 inhibitor (e.g. dinaciclib), a MCL-1 inhibitor, temozolomide, a Bcl-2 inhibitor (e.g. venetoclax), a Bcl-xL inhibitor, ibrutinib, a mTOR inhibitor (e.g. everolimus), a PI3K inhibitor (e.g. buparlisib), duvelisib, idelalisib, an AKT inhibitor, a HER2 inhibitor (e.g. lapatinib), a taxane (e.g. docetaxel, paclitaxel, nab-paclitaxel), an ADC comprising an auristatin, an ADC comprising a PBD (e.g. rovalpituzumab tesirine), an ADC comprising a maytansinoid (e.g. TDM1), a TRAIL agonist, a proteasome inhibitor (e.g. bortezomib), and a nicotinamide phosphoribosyltransferase (NAMPT) inhibitor.

In one embodiment, the anti-B7-H3 ADCs of the invention are administered in combination with venetoclax to a human subject for the treatment of small cell lung cancer (SCLC).

In one embodiment, the additional therapy is radiation.

In one embodiment, the additional agent is a chemotherapeutic agent.

In one aspect, the present invention provides a process for the preparation of an ADC according to structural formula (I):

(D-L-LK m Ab (I)

wherein:

D is the Bcl-xL inhibitor drug of formula (IIa), (IIb), (IIc), or (IId) as disclosed herein;

L is the linker as disclosed herein;

Ab is an hB7-H3 antibody, wherein the hB7-H3 antibody comprises the heavy and light chain CDRs of huAb3v2.5, huAb3v2.6, or huAb13v1;

LK represents a covalent linkage linking linker L to antibody Ab; and

m is an integer ranging from 1 to 20;

the process comprising:

treating an antibody in an aqueous solution with an effective amount of a disulfide reducing agent at 30-40° C. for at least 15 minutes, and then cooling the antibody solution to 20-27° C.;

adding to the reduced antibody solution a solution of water/dimethyl sulfoxide comprising a synthon selected from the group of 2.1 to 2.176 (Table B);

adjusting the pH of the solution to a pH of 7.5 to 8.5;

allowing the reaction to run for 48 to 80 hours to form the ADC;

wherein the mass is shifted by 18±2 amu for each hydrolysis of a succinimide to a succinamide as measured by electron spray mass spectrometry; and

wherein the ADC is optionally purified by hydrophobic interaction chromatography.

In one embodiment, m is 2.

In another aspect, the present invention provides an ADC prepared by the process as described above.

›BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graphical representation of the epitope grouping of murine anti-B7-H3 hybridoma antibodies as determined by pair-wise binding assays.

FIG. 2 depicts an antibody reduction, modification with a maleimide derivative to give a thiosuccinimide intermediate, and subsequent hydrolysis of thiosuccinimide moiety FIG. 3 depicts the structure of an antibody-maleimidocaproyl-vc-PABA-MMAE ADC.

FIG. 4 depicts the structure of a PBD dimer (SGD-1882) conjugated to an antibody (Ab) via a maleimidocaproyl-valine-alanine linker (collectively referred to as SGD-1910).

FIG. 5 depicts the MS characterization of light chain and heavy chain of huAb13v1 1) prior to conjugation, 2) after conjugation to a maleimide derivative to give a thiosuccinimide intermediate and 3) post pH 8-mediated hydrolysis of the thiosuccinimide ring.

›DETAILED DESCRIPTION OF THE INVENTION · 1 of 40

Various aspects of the invention relate to anti-B7-H3 antibodies and antibody fragments, anti-B7-H3 ADCs, and pharmaceutical compositions thereof, as well as nucleic acids, recombinant expression vectors and host cells for making such antibodies and fragments. Methods of using the antibodies, fragments, and ADCs described herein to detect human B7-H3, to inhibit human B7-H3 activity (in vitro or in vivo), and to treat cancers are also encompassed by the invention. In certain embodiments, the invention provides anti-B7-H3 ADCs, including ADCs comprising Bcl-xL inhibitors, synthons useful for synthesizing the ADCs, compositions comprising the ADCs, methods of making the ADCs, and various methods of using the ADCs.

As will be appreciated by skilled artisans, the ADCs disclosed herein are “modular” in nature. Throughout the instant disclosure, various specific embodiments of the various “modules” comprising the ADCs, as well as the synthons useful for synthesizing the ADCs, are described. As specific non-limiting examples, specific embodiments of antibodies, linkers, and Bcl-xL inhibitors that may comprise the ADCs and synthons are described. It is intended that all of the specific embodiments described may be combined with each other as though each specific combination were explicitly described individually.

It will also be appreciated by skilled artisans that the various ADCs and/or ADC synthons described herein may be in the form of salts, and in certain embodiments, particularly pharmaceutically acceptable salts. The compounds of the present disclosure that possess a sufficiently acidic, a sufficiently basic, or both functional groups, can react with any of a number of inorganic bases, and inorganic and organic acids, to form a salt. Alternatively, compounds that are inherently charged, such as those with a quaternary nitrogen, can form a salt with an appropriate counterion, e.g., a halide such as a bromide, chloride, or fluoride.

Acids commonly employed to form acid addition salts are inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, and the like, and organic acids such as p-toluenesulfonic acid, methanesulfonic acid, oxalic acid, p-bromophenyl-sulfonic acid, carbonic acid, succinic acid, citric acid, etc. Base addition salts include those derived from inorganic bases, such as ammonium and alkali or alkaline earth metal hydroxides, carbonates, bicarbonates, and the like.

In the disclosure below, if both structural diagrams and nomenclature are included and if the nomenclature conflicts with the structural diagram, the structural diagram controls.

An outline of the Detailed Description of the Invention is provided below:

I. Definitions

II. Anti-B7-H3 Antibodies

II.A. Anti-B7-H3 Chimeric Antibodies

II.B. Humanized Anti-B7-H3 Antibodies

III. Anti-B7-H3 Antibody Drug Conjugates (ADCs)

III.A. Anti-B7-H3/Bcl-xL Inhibitor ADCs

III.A.1. Bcl-xL Inhibitors III.A.2 Bcl-xL Linkers

Cleavable Linkers Non-Cleavable Linkers Groups Used to Attach Linkers to Anti-B7-H3 Antibodies Linker Selection Considerations

III.A.3. Bcl-xL ADC Synthons III.A.4 Methods of Synthesis of Bcl-xL ADCs III.A.5. General Methods for Synthesizing Bcl-xL Inhibitors III.A.6. General Methods for Synthesizing Synthons III.A.7. General Methods for Synthesizing Anti-B7-H3 ADCs

III.B. Anti-B7-H3 ADCs: Other Exemplary Drugs for Conjugation

III.C. Anti-B7-H3 ADCs: Other Exemplary Linkers

IV. Purification of Anti-B7-H3 ADCs

V. Uses of Anti-B7-H3 Antibodies and Anti-B7-H3 ADCs

VI. Pharmaceutical Compositions

I. Definitions

In order that the invention may be more readily understood, certain terms are first defined. In addition, it should be noted that whenever a value or range of values of a parameter are recited, it is intended that values and ranges intermediate to the recited values are also intended to be part of this invention.

The term “anti-B7-H3 antibody” refers to an antibody that specifically binds to B7-H3. An antibody “which binds” an antigen of interest, i.e., B7-H3, is one capable of binding that antigen with sufficient affinity such that the antibody is useful in targeting a cell expressing the antigen. In a preferred embodiment, the antibody specifically binds to human B7-H3 (hB7-H3). Examples of anti-B7-H3 antibodies are disclosed in the examples below. Unless otherwise indicated, the term “anti-B7-H3 antibody” is meant to refer to an antibody which binds to wild type B7-H3 (e.g., a 4IgB7-H3 isoform of B7-H3) or any variant of B7-H3. The amino acid sequence of wild type human B7-H3 is provided below as SEQ ID NO: 149, where the signal peptide (amino acid residues 1-28) is underlined.

(SEQ ID NO: 149) MLRRRGSPGMGVHVGAALGALWFC LTGALEVQVPEDPVVALVGTDAT LCCSFSPEPGFSLAQLNLIWQLTDTKQLVHSFAEGQDQGSAYANRTA LFPDLLAQGNASLRLQRVRVADEGSFTCFVSIRDFGSAAVSLQVAAP YSKPSMTLEPNKDLRPGDTVTITCSSYQGYPEAEVFWQDGQGVPLTG NVTTSQMANEQGLFDVHSILRVVLGANGTYSCLVRNPVLQQDAHSSV TITPQRSPTGAVEVQVPEDPVVALVGTDATLRCSFSPEPGESLAQLN LIWQLTDTKQLVHSFTEGRDQGSAYANRTALFPDLLAQGNASLRLQR VRVADEGSFTCFVSIRDFGSAAVSLQVAAPYSKPSMTLEPNKDLRPG DTVTITCSSYRGYPEAEVFWQDGQGVPLTGNVTTSQMANEQGLFDVH SVLRVVLGANGTYSCLVRNPVLQQDAHGSVTITGQPMTFPPEALWVT VGLSVCLIALLVALAFVCWRKIKQSCEEENAGAEDQDGEGEGSKTAL QPLKHSDSKEDDGQEIA

Thus, in one embodiment of the invention, the antibody or ADC binds human B7-H3 as defined in SEQ ID NO: 149. The extracellular domain (ECD) of human B7-H3 is provided in SEQ ID NO: 152 (inclusive of a His tag). As such, in one embodiment of the invention, the antibody of ADC binds the ECD of human B7-H3 as described in the ECD of SEQ ID NO: 152.

The terms “specific binding” or “specifically binding”, as used herein, in reference to the interaction of an antibody or an ADC with a second chemical species, mean that the interaction is dependent upon the presence of a particular structure (e.g., an antigenic determinant or epitope) on the chemical species; for example, an antibody recognizes and binds to a specific protein structure rather than to proteins generally. If an antibody or ADC is specific for epitope “A”, the presence of a molecule containing epitope A (or free, unlabeled A), in a reaction containing labeled “A” and the antibody, will reduce the amount of labeled A bound to the antibody or ADC. By way of example, an antibody “binds specifically” to a target if the antibody, when labeled, can be competed away from its target by the corresponding non-labeled antibody. In one embodiment, an antibody specifically binds to a target, e.g., B7-H3, if the antibody has a K D for the target of at least about 10 −4 M, 10 −5 M, 10 −6 M, 10-7 M, 10 −8 M, 10 −9 M, 10 −10 M, 10 −11 M, 10 −12 M, or less (less meaning a number that is less than 10 −12 , e.g. 10 −13 ). In one embodiment, the term “specific binding to B7-H3” or “specifically binds to B7-H3,” as used herein, refers to an antibody or an ADC that binds to B7-H3 and has a dissociation constant (K D ) of 1.0×10 −7 M or less, as determined by surface plasmon resonance. It shall be understood, however, that the antibody or ADC may be capable of specifically binding to two or more antigens which are related in sequence. For example, in one embodiment, an antibody can specifically bind to both human and a non-human (e.g., mouse or non-human primate) orthologs of B7-H3.

›DETAILED DESCRIPTION OF THE INVENTION · 2 of 40

The term “antibody” refers to an immunoglobulin molecule that specifically binds to an antigen and comprises a heavy (H) chain(s) and a light (L chain(s). Each heavy chain is comprised of a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region. The heavy chain constant region is comprised of three domains, CH1, CH2 and CH3. Each light chain is comprised of a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region is comprised of one domain, CL. The VH and VL regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDR), interspersed with regions that are more conserved, termed framework regions (FR). Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. An antibody can be of any type (e.g., IgG, IgE, IgM, IgD, IgA and IgY) and class (e.g., IgG1, IgG2, IgG 3, IgG4, IgA1 and IgA2) or subclass.

While the term “antibody” is not intended to include antigen binding portions of an antibody (defined below), it is intended, in certain embodiments, to include a small number of amino acid deletions from the carboxy end of the heavy chain(s). In one embodiment, an antibody comprises a heavy chain having 1-5 amino acid deletions the carboxy end of the heavy chain. In one embodiment, an antibody is a monoclonal antibody which is an IgG, having four polypeptide chains, two heavy (H) chains, and two light (L chains) that can bind to hB7-H3. In one embodiment, an antibody is a monoclonal IgG antibody comprising a lambda or a kappa light chain.

The term “antigen binding portion” or “antigen binding fragment” of an antibody (or simply “antibody portion” or “antibody fragment”), as used herein, refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen (e.g., hB7-H3). It has been shown that the antigen binding function of an antibody can be performed by fragments of a full-length antibody. Such antibody embodiments may also be bispecific, dual specific, or multi-specific formats; specifically binding to two or more different antigens. Examples of binding fragments encompassed within the term “antigen binding portion” of an antibody include (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CH1 domains; (ii) a F(ab′) 2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of the VH and CH1 domains; (iv) a Fv fragment consisting of the VL and VH domains of a single arm of an antibody, (v) a dAb fragment (Ward et al., (1989) Nature 341:544-546, Winter et al., PCT publication WO 90/05144 A1 herein incorporated by reference), which comprises a single variable domain; and (vi) an isolated complementarity determining region (CDR). Furthermore, although the two domains of the Fv fragment, VL and VH, are coded for by separate genes, they can be joined, using recombinant methods, by a synthetic linker that enables them to be made as a single protein chain in which the VL and VH regions pair to form monovalent molecules (known as single chain Fv (scFv); see e.g., Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883). Such single chain antibodies are also intended to be encompassed within the term “antigen binding portion” of an antibody. In certain embodiments of the invention, scFv molecules may be incorporated into a fusion protein. Other forms of single chain antibodies, such as diabodies are also encompassed. Diabodies are bivalent, bispecific antibodies in which VH and VL domains are expressed on a single polypeptide chain, but using a linker that is too short to allow for pairing between the two domains on the same chain, thereby forcing the domains to pair with complementary domains of another chain and creating two antigen binding sites (see e.g., Holliger, P., et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak, R. J., et al. (1994) Structure 2:1121-1123). Such antibody binding portions are known in the art (Kontermann and Dubel eds., Antibody Engineering (2001) Springer-Verlag. New York. 790 pp. (ISBN 3-540-41354-5).

An IgG is a class of antibody comprising two heavy chains and two light chains arranged in a Y-shape. An IgG constant domain refers to a heavy or light chain constant domain. Exemplary human IgG heavy chain and light chain constant domain amino acid sequences are known in the art and represented below in Table A.

An “isolated antibody”, as used herein, is intended to refer to an antibody that is substantially free of other antibodies having different antigenic specificities (e.g., an isolated antibody that specifically binds B7-H3 is substantially free of antibodies that specifically bind antigens other than B7-H3). An isolated antibody that specifically binds B7-H3 may, however, have cross-reactivity to other antigens, such as B7-H3 molecules from other species. Moreover, an isolated antibody may be substantially free of other cellular material and/or chemicals.

The term “humanized antibody” refers to antibodies which comprise heavy and light chain variable region sequences from a nonhuman species (e.g., a mouse) but in which at least a portion of the VH and/or VL sequence has been altered to be more “human-like”, i.e., more similar to human germline variable sequences. In particular, the term “humanized antibody” is an antibody or a variant, derivative, analog or fragment thereof which immunospecifically binds to an antigen of interest and which comprises a framework (FR) region having substantially the amino acid sequence of a human antibody and a complementary determining region (CDR) having substantially the amino acid sequence of a non-human antibody. As used herein, the term “substantially” in the context of a CDR refers to a CDR having an amino acid sequence at least 80%, preferably at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to the amino acid sequence of a non-human antibody CDR. A humanized antibody comprises substantially all of at least one, and typically two, variable domains (Fab, Fab′, F(ab′) 2 , FabC, Fv) in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin (i.e., donor antibody) and all or substantially all of the framework regions are those of a human immunoglobulin consensus sequence. Preferably, a humanized antibody also comprises at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. In some embodiments, a humanized antibody contains both the light chain as well as at least the variable domain of a heavy chain. The antibody also may include the CH1, hinge, CH2, CH3, and CH4 regions of the heavy chain. In some embodiments, a humanized antibody only contains a humanized light chain. In other embodiments, a humanized antibody only contains a humanized heavy chain. In specific embodiments, a humanized antibody only contains a humanized variable domain of a light chain and/or humanized heavy chain.

›DETAILED DESCRIPTION OF THE INVENTION · 3 of 40

The humanized antibody can be selected from any class of immunoglobulins, including IgM, IgG, IgD, IgA and IgE, and any isotype, including without limitation IgG1, IgG2, IgG3 and IgG4. The humanized antibody may comprise sequences from more than one class or isotype, and particular constant domains may be selected to optimize desired effector functions using techniques well-known in the art.

The terms “Kabat numbering,” “Kabat definitions,” and “Kabat labeling” are used interchangeably herein. These terms, which are recognized in the art, refer to a system of numbering amino acid residues which are more variable (i.e., hypervariable) than other amino acid residues in the heavy and light chain variable regions of an antibody, or an antigen binding portion thereof (Kabat et al. (1971) Ann. NY Acad, Sci. 190:382-391 and, Kabat, E. A., et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition , U.S. Department of Health and Human Services, NIH Publication No. 91-3242). For the heavy chain variable region, the hypervariable region ranges from amino acid positions 31 to 35 for CDR1, amino acid positions 50 to 65 for CDR2, and amino acid positions 95 to 102 for CDR3. For the light chain variable region, the hypervariable region ranges from amino acid positions 24 to 34 for CDR1, amino acid positions 50 to 56 for CDR2, and amino acid positions 89 to 97 for CDR3.

As used herein, the term “CDR” refers to the complementarity determining region within antibody variable sequences. There are three CDRs in each of the variable regions of the heavy chain (HC) and the light chain (LC), which are designated CDR1, CDR2 and CDR3 (or specifically HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3), for each of the variable regions. The term “CDR set” as used herein refers to a group of three CDRs that occur in a single variable region capable of binding the antigen. The exact boundaries of these CDRs have been defined differently according to different systems. The system described by Kabat (Kabat et al., Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987) and (1991)) not only provides an unambiguous residue numbering system applicable to any variable region of an antibody, but also provides precise residue boundaries defining the three CDRs. These CDRs may be referred to as Kabat CDRs. Chothia and coworkers (Chothia & Lesk, J. Mol. Biol. 196:901-917 (1987) and Chothia et al., Nature 342:877-883 (1989)) found that certain sub-portions within Kabat CDRs adopt nearly identical peptide backbone conformations, despite having great diversity at the level of amino acid sequence. These sub-portions were designated as L1, L2 and L3 or H1, H2 and H3 where the “L” and the “H” designates the light chain and the heavy chains regions, respectively. These regions may be referred to as Chothia CDRs, which have boundaries that overlap with Kabat CDRs. Other boundaries defining CDRs overlapping with the Kabat CDRs have been described by Padlan (FASEB J. 9:133-139 (1995)) and MacCallum (J. Mol. Biol. 262(5):732-45 (1996)). Still other CDR boundary definitions may not strictly follow one of the above systems, but will nonetheless overlap with the Kabat CDRs, although they may be shortened or lengthened in light of prediction or experimental findings that particular residues or groups of residues or even entire CDRs do not significantly impact antigen binding. The methods used herein may utilize CDRs defined according to any of these systems, although preferred embodiments use Kabat or Chothia defined CDRs.

As used herein, the term “framework” or “framework sequence” refers to the remaining sequences of a variable region minus the CDRs. Because the exact definition of a CDR sequence can be determined by different systems, the meaning of a framework sequence is subject to correspondingly different interpretations. The six CDRs (CDR-L1, CDR-L2, and CDR-L3 of light chain and CDR-H1, CDR-H2, and CDR-H3 of heavy chain) also divide the framework regions on the light chain and the heavy chain into four sub-regions (FR1, FR2, FR3 and FR4) on each chain, in which CDR1 is positioned between FR1 and FR2, CDR2 between FR2 and FR3, and CDR3 between FR3 and FR4. Without specifying the particular sub-regions as FR1, FR2, FR3 or FR4, a framework region, as referred by others, represents the combined FR's within the variable region of a single, naturally occurring immunoglobulin chain. As used herein, a FR represents one of the four sub-regions, and FRs represents two or more of the four sub-regions constituting a framework region.

The framework and CDR regions of a humanized antibody need not correspond precisely to the parental sequences, e.g., the donor antibody CDR or the consensus framework may be mutagenized by substitution, insertion and/or deletion of at least one amino acid residue so that the CDR or framework residue at that site does not correspond to either the donor antibody or the consensus framework. In a preferred embodiment, such mutations, however, will not be extensive. Usually, at least 80%, preferably at least 85%, more preferably at least 90%, and most preferably at least 95% of the humanized antibody residues will correspond to those of the parental FR and CDR sequences. As used herein, the term “consensus framework” refers to the framework region in the consensus immunoglobulin sequence. As used herein, the term “consensus immunoglobulin sequence” refers to the sequence formed from the most frequently occurring amino acids (or nucleotides) in a family of related immunoglobulin sequences (See e.g., Winnaker, From Genes to Clones (Verlagsgesellschaft, Weinheim, Germany 1987). In a family of immunoglobulins, each position in the consensus sequence is occupied by the amino acid occurring most frequently at that position in the family. If two amino acids occur equally frequently, either can be included in the consensus sequence.

The term “human acceptor framework”, as used herein, is meant to refer to a framework of an antibody or antibody fragment thereof comprising the amino acid sequence of a VH or VL framework derived from a human antibody or antibody fragment thereof or a human consensus sequence framework into which CDR's from a non-human species may be incorporated.

›DETAILED DESCRIPTION OF THE INVENTION · 4 of 40

“Percent (%) amino acid sequence identity” with respect to a peptide or polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical with the amino acid residues in the specific peptide or polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in various ways that are within the skill in the art, for instance, using publicly available computer software such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared. In one embodiment, the invention includes an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to an amino acid sequence set forth in any one of SEQ ID NOs: 1 to 148.

The term “multivalent antibody” is used herein to denote an antibody comprising two or more antigen binding sites. In certain embodiments, the multivalent antibody may be engineered to have the three or more antigen binding sites, and is generally not a naturally occurring antibody.

The term “multispecific antibody” refers to an antibody capable of binding two or more unrelated antigens. In one embodiment, the multispecific antibody is a bispecific antibody that is capable of binding to two unrelated antigens, e.g., a bispecific antibody, or antigen-binding portion thereof, that binds B7-H3 and CD3.

The term “dual variable domain” or “DVD,” as used interchangeably herein, are antigen binding proteins that comprise two or more antigen binding sites and are tetravalent or multivalent binding proteins. Such DVDs may be monospecific, i.e., capable of binding one antigen or multispecific, i.e. capable of binding two or more antigens. DVD binding proteins comprising two heavy chain DVD polypeptides and two light chain DVD polypeptides are referred to a DVD Ig. Each half of a DVD Ig comprises a heavy chain DVD polypeptide, and a light chain DVD polypeptide, and two antigen binding sites. Each binding site comprises a heavy chain variable domain and a light chain variable domain with a total of 6 CDRs involved in antigen binding per antigen binding site. In one embodiment, the CDRs described herein are used in an anti-B7-H3 DVD.

The term “chimeric antigen receptor” or “CAR” refers to a recombinant protein comprising at least (1) an antigen-binding region, e.g., a variable heavy or light chain of an antibody, (2) a transmembrane domain to anchor the CAR into a T cell, and (3) one or more intracellular signaling domains.

The term “activity” includes activities such as the binding specificity/affinity of an antibody or ADC for an antigen, for example, an anti-hB7-H3 antibody that binds to an hB7-H3 antigen and/or the neutralizing potency of an antibody, for example, an anti-hB7-H3 antibody whose binding to hB7-H3 inhibits the biological activity of hB7-H3, e.g., inhibition of proliferation of B7-H3 expressing cell lines, e.g., human H146 lung carcinoma cells, human H1650 lung carcinoma cells, or human EBC1 lung carcinoma cells.

The term “non small-cell lung carcinoma (NSCLC) xenograft assay,” as used herein, refers to an in vivo assay used to determine whether an anti-B7-H3 antibody or ADC, can inhibit tumor growth (e.g., further growth) and/or decrease tumor growth resulting from the transplantation of NSCLC cells into an immunodeficient mouse. An NSCLC xenograft assay includes transplantation of NSCLC cells into an immunodeficient mouse such that a tumor grows to a desired size, e.g., 200-250 mm 3 , whereupon the antibody or ADC is administered to the mouse to determine whether the antibody or ADC can inhibit and/or decrease tumor growth. In certain embodiments, the activity of the antibody or ADC is determined according to the percent tumor growth inhibition (% TGI) relative to a control antibody, e.g., a human IgG antibody (or collection thereof) which does not specifically bind tumor cells, e.g., is directed to an antigen not associated with cancer or is obtained from a source which is noncancerous (e.g., normal human serum). In such embodiments, the antibody (or ADC) and the control antibody are administered to the mouse at the same dose, with the same frequency, and via the same route. In one embodiment, the mouse used in the NSCLC xenograft assay is a severe combined immunodeficiency (SCID) mouse and/or an athymic CD-1 nude mouse. Examples of NSCLC cells that may be used in the NSCLC xenograft assay include, but are not limited to, H1299 cells (NCI-H1299 [H-1299] (ATCC® CRL-5803)), H1975 cells (NCI-H1975 cells [H1975] (ATCC® CRL-5908™)), and EBC-1 cells.

The term “small-cell lung carcinoma (SCLC) xenograft assay,” as used herein, refers to an in vivo assay used to determine whether an anti-B7-H3 antibody or ADC, can inhibit tumor growth (e.g., further growth) and/or decrease tumor growth resulting from the transplantation of SCLC cells into an immunodeficient mouse. An SCLC xenograft assay includes transplantation of SCLC cells into an immunodeficient mouse such that a tumor grows to a desired size, e.g., 200-250 mm 3 , whereupon the antibody or ADC is administered to the mouse to determine whether the antibody or ADC can inhibit and/or decrease tumor growth. In certain embodiments, the activity of the antibody or ADC is determined according to the percent tumor growth inhibition (% TGI) relative to a control antibody, e.g., a human IgG antibody (or collection thereof) which does not specifically bind tumor cells, e.g., is directed to an antigen not associated with cancer or is obtained from a source which is noncancerous (e.g., normal human serum). In such embodiments, the antibody (or ADC) and the control antibody are administered to the mouse at the same dose, with the same frequency, and via the same route. In one embodiment, the mouse used in the NSCLC xenograft assay is a severe combined immunodeficiency (SCID) mouse and/or an athymic CD-1 nude mouse. Examples of SCLC cells that may be used in the SCLC xenograft assay include, but are not limited to, H146 cells (NCI-H146 cells [H146] (ATCC® HTB-173™)), and H847 cells (NCI-H847 [H847] (ATCC® CRL-5846™)). The term “epitope” refers to a region of an antigen that is bound by an antibody or ADC. In certain embodiments, epitope determinants include chemically active surface groupings of molecules such as amino acids, sugar side chains, phosphoryl, or sulfonyl, and, in certain embodiments, may have specific three dimensional structural characteristics, and/or specific charge characteristics. In certain embodiments, an antibody is said to specifically bind an antigen when it preferentially recognizes its target antigen in a complex mixture of proteins and/or macromolecules.

›DETAILED DESCRIPTION OF THE INVENTION · 5 of 40

The term “surface plasmon resonance”, as used herein, refers to an optical phenomenon that allows for the analysis of real-time biospecific interactions by detection of alterations in protein concentrations within a biosensor matrix, for example using the BIAcore system (Pharmacia Biosensor AB, Uppsala, Sweden and Piscataway, N.J.). For further descriptions, see Jönsson, U., et al. (1993) Ann. Biol. Clin. 51:19-26; Jönsson, U., et al. (1991) Biotechniques 11:620-627; Johnsson, B., et al. (1995) J. Mol. Recognit. 8:125-131; and Johnnson, B., et al. (1991) Anal. Biochem. 198:268-277. In one embodiment, surface plasmon resonance is determined according to the methods described in Example 2.

The term “k on ” or “k a ”, as used herein, is intended to refer to the on rate constant for association of an antibody to the antigen to form the antibody/antigen complex.

The term “k off ” or “k d ”, as used herein, is intended to refer to the off rate constant for dissociation of an antibody from the antibody/antigen complex.

The term “K D ”, as used herein, is intended to refer to the equilibrium dissociation constant of a particular antibody-antigen interaction (e.g., huAb13 antibody and B7-H3). K D is calculated by k a /k d .

The term “competitive binding”, as used herein, refers to a situation in which a first antibody competes with a second antibody, for a binding site on a third molecule, e.g., an antigen. In one embodiment, competitive binding between two antibodies is determined using FACS analysis.

The term “competitive binding assay” is an assay used to determine whether two or more antibodies bind to the same epitope. In one embodiment, a competitive binding assay is a competition fluorescent activated cell sorting (FACS) assay which is used to determine whether two or more antibodies bind to the same epitope by determining whether the fluorescent signal of a labeled antibody is reduced due to the introduction of a non-labeled antibody, where competition for the same epitope will lower the level of fluorescence.

The term “labeled antibody” as used herein, refers to an antibody, or an antigen binding portion thereof, with a label incorporated that provides for the identification of the binding protein, e.g., an antibody. Preferably, the label is a detectable marker, e.g., incorporation of a radiolabeled amino acid or attachment to a polypeptide of biotinyl moieties that can be detected by marked avidin (e.g., streptavidin containing a fluorescent marker or enzymatic activity that can be detected by optical or colorimetric methods). Examples of labels for polypeptides include, but are not limited to, the following: radioisotopes or radionuclides (e.g., 3 H, 14 C, 35 S, 99 Y, 99 Tc, 111 In, 125 I, 131 I, 177 Lu, 166 Ho, or 153 Sm); fluorescent labels (e.g., FITC, rhodamine, lanthanide phosphors), enzymatic labels (e.g., horseradish peroxidase, luciferase, alkaline phosphatase); chemiluminescent markers; biotinyl groups; predetermined polypeptide epitopes recognized by a secondary reporter (e.g., leucine zipper pair sequences, binding sites for secondary antibodies, metal binding domains, epitope tags); and magnetic agents, such as gadolinium chelates.

The term “antibody-drug-conjugate” or “ADC” refers to a binding protein, such as an antibody or antigen binding fragment thereof, chemically linked to one or more chemical drug(s) (also referred to herein as agent(s), warhead(s), and payload(s)) that may optionally be therapeutic or cytotoxic agents. In a preferred embodiment, an ADC includes an antibody, a drug, (e.g. a cytotoxic drug), and a linker that enables attachment or conjugation of the drug to the antibody. An ADC typically has anywhere from 1 to 8 drugs conjugated to the antibody, including drug loaded species of 2, 4, 6, or 8. Non-limiting examples of drugs that may be included in the ADCs are mitotic inhibitors, antitumor antibiotics, immunomodulating agents, vectors for gene therapy, alkylating agents, antiangiogenic agents, antimetabolites, boron-containing agents, chemoprotective agents, hormones, antihormone agents, corticosteroids, photoactive therapeutic agents, oligonucleotides, radionuclide agents, topoisomerase inhibitors, kinase inhibitors (e.g., TEC-family kinase inhibitors and serine/threonine kinase inhibitors), and radiosensitizers. In one embodiment, the drug is a Bcl-xL inhibitor.

The terms “anti-B7-H3 antibody drug conjugate” or “anti-B7-H3 ADC”, used interchangeably herein, refer to an ADC comprising an antibody that specifically binds to B7-H3, whereby the antibody is conjugated to one or more chemical agent(s). In one embodiment, the anti-B7-H3 ADC comprises antibody huAb13v1, huAb3v2.5, or huAb3v2.6 conjugated to an auristatin, e.g., MMAE or MMAF. In one embodiment, the anti-B7-H3 ADC comprises antibody huAb13v1, huAb3v2.5, or huAb3v2.6 conjugated to a Bcl-xL inhibitor. In a preferred embodiment, the anti-B7-H3 ADC binds to human B7-H3 (hB7-H3).

The term “Bcl-xL inhibitor”, as used herein, refers to a compound which antagonizes Bcl-xL activity in a cell. In one embodiment, a Bcl-xL inhibitor promotes apoptosis of a cell by inhibiting Bcl-xL activity.

The term “auristatin”, as used herein, refers to a family of antimitotic agents. Auristatin derivatives are also included within the definition of the term “auristatin”. Examples of auristatins include, but are not limited to, auristatin E (AE), monomethylauristatin E (MMAE), monomethylauristatin F (MMAF), and synthetic analogs of dolastatin. In one embodiment, an anti-B7-H3 antibody described herein is conjugated to an auristatin to form an anti-B7-H3 ADC.

As used herein, the term “Ab-vcMMAE” is used to refer to an ADC comprising an antibody conjugated to monomethylauristatin E (MMAE) via a maleimidocaproyl valine citrulline p-aminobenzyloxycarbamyl (PABA) linker.

As used herein, the term “mcMMAF” is used to refer to a linker/drug combination of maleimidocaproyl-monomethylauristatin F (MMAF).

The term “drug-to-antibody ratio” or “DAR” refers to the number of drugs, e.g., Bcl-xL inhibitor, attached to the antibody of the ADC. The DAR of an ADC can range from 1 to 8, although higher loads, e.g., 20, are also possible depending on the number of linkage site on an antibody. The term DAR may be used in reference to the number of drugs loaded onto an individual antibody, or, alternatively, may be used in reference to the average or mean DAR of a group of ADCs.

›DETAILED DESCRIPTION OF THE INVENTION · 6 of 40

The term “undesired ADC species”, as used herein, refers to any drug loaded species which is to be separated from an ADC species having a different drug load. In one embodiment, the term undesired ADC species may refer to drug loaded species of 6 or more, i.e., ADCs with a DAR of 6 or more, including DAR6, DAR7, DAR8, and DAR greater than 8 (i.e., drug loaded species of 6, 7, 8, or greater than 8). In a separate embodiment, the term undesired ADC species may refer to drug loaded species of 8 or more, i.e., ADCs with a DAR of 8 or more, including DAR8, and DAR greater than 8 (i.e., drug loaded species of 8, or greater than 8).

The term “ADC mixture”, as used herein, refers to a composition containing a heterogeneous DAR distribution of ADCs. In one embodiment, an ADC mixture contains ADCs having a distribution of DARs of 1 to 8, e.g., 1.5, 2, 4, 6, and 8 (i.e., drug loaded species of 2, 4, 6, and 8). Notably, degradation products may result such that DARs of 1, 3, 5, and 7 may also be included in the mixture. Further, ADCs within the mixture may also have DARs greater than 8. The ADC mixture results from interchain disulfide reduction followed by conjugation. In one embodiment, the ADC mixture comprises both ADCs with a DAR of 4 or less (i.e., a drug loaded species of 4 or less) and ADCs with a DAR of 6 or more (i.e., a drug loaded species of 6 or more).

The term a “xenograft assay”, as used herein, refers to a human tumor xenograft assay, wherein human tumor cells are transplanted, either under the skin or into the organ type in which the tumor originated, into immunocompromised mice that do not reject human cells.

The term “cancer” is meant to refer to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth. Examples of cancer include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia or lymphoid malignancies. More particular examples of such cancers include glioblastoma, acute myeloid leukemia (AML), non-Hodgkin's lymphoma (NHL), non-small cell lung cancer, lung cancer, colon cancer, colorectal cancer, head and neck cancer, breast cancer (e.g., triple negative breast cancer), pancreatic cancer, squamous cell tumors, squamous cell carcinoma (e.g., squamous cell lung cancer or squamous cell head and neck cancer), anal cancer, skin cancer, and vulvar cancer. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having a tumor(s) that overexpresses B7-H3. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having a solid tumor which is likely to over-express B7-H3. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having squamous cell non-small cell lung cancer (NSCLC). In one embodiment, the antibodies or ADCs of the invention are administered to a patient having solid tumors, including advanced solid tumors. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having prostate cancer. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having non-small cell lung cancer. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having a glioblastoma. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having colon cancer. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having head and neck cancer. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having kidney cancer. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having clear cell renal cell carcinoma. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having glioma. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having melanoma. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having pancreatic cancer. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having gastric cancer. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having ovarian cancer. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having colorectal cancer. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having renal cancer. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having small cell lung cancer. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having hepatocellular carcinoma. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having hypopharyngeal squamous cell carcinoma. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having neuroblastoma. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having breast cancer. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having endometrial cancer. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having urothelial cell carcinoma. In one embodiment, the antibodies or ADCs of the invention are administered to a patient having acute myeloid leukemia (AML). In one embodiment, the antibodies or ADCs of the invention are administered to a patient having non-Hodgkin's lymphoma (NHL).

The term “B7-H3 expressing tumor,” as used herein, refers to a tumor which expresses B7-H3 protein. In one embodiment, B7-H3 expression in a tumor is determined using immunohistochemical staining of tumor cell membranes, where any immunohistochemical staining above background level in a tumor sample indicates that the tumor is a B7-H3 expressing tumor. Methods for detecting expression of B7-H3 in a tumor are known in the art, and include immunohistochemical assays. In contrast, a “B7-H3 negative tumor” is defined as a tumor having an absence of B7-H3 membrane staining above background in a tumor sample as determined by immunohistochemical techniques.

›DETAILED DESCRIPTION OF THE INVENTION · 7 of 40

The terms “overexpress,” “overexpression,” or “overexpressed” interchangeably refer to a gene that is transcribed or translated at a detectably greater level, usually in a cancer cell, in comparison to a normal cell. Overexpression therefore refers to both overexpression of protein and RNA (due to increased transcription, post transcriptional processing, translation, post translational processing, altered stability, and altered protein degradation), as well as local overexpression due to altered protein traffic patterns (increased nuclear localization), and augmented functional activity, e.g., as in an increased enzyme hydrolysis of substrate. Thus, overexpression refers to either protein or RNA levels. Overexpression can also be by 50%, 60%, 70%, 80%, 90% or more in comparison to a normal cell or comparison cell. In certain embodiments, the anti-B7-H3 antibodies or ADCs of the invention are used to treat solid tumors likely to overexpress B7-H3.

The term “gene amplification”, as used herein, refers to a cellular process characterized by the production of multiple copies of any particular piece of DNA. For example, a tumor cell may amplify, or copy, chromosomal segments as a result of cell signals and sometimes environmental events. The process of gene amplification leads to the production of additional copies of the gene. In one embodiment, the gene is B7-H3, i.e., “B7-H3 amplification.” In one embodiment, the compositions and methods disclosed herein are used to treat a subject having B7-H3 amplified cancer.

The term “administering” as used herein is meant to refer to the delivery of a substance (e.g., an anti-B7-H3 antibody or ADC) to achieve a therapeutic objective (e.g., the treatment of a B7-H3-associated disorder). Modes of administration may be parenteral, enteral and topical. Parenteral administration is usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion.

The term “combination therapy” or “combination” in the context of a therapeutic method (e.g., a treatment method), as used herein, refers to the administration of two or more therapeutic substances, e.g., an anti-B7-H3 antibody or ADC and an additional therapeutic agent. The additional therapeutic agent may be administered concomitant with, prior to, or following the administration of the anti-B7-H3 antibody or ADC.

As used herein, the term “effective amount” or “therapeutically effective amount” refers to the amount of a drug, e.g., an antibody or ADC, which is sufficient to reduce or ameliorate the severity and/or duration of a disorder, e.g., cancer, or one or more symptoms thereof, prevent the advancement of a disorder, cause regression of a disorder, prevent the recurrence, development, onset or progression of one or more symptoms associated with a disorder, detect a disorder, or enhance or improve the prophylactic or therapeutic effect(s) of another therapy (e.g., prophylactic or therapeutic agent). The effective amount of an antibody or ADC may, for example, inhibit tumor growth (e.g., inhibit an increase in tumor volume), decrease tumor growth (e.g., decrease tumor volume), reduce the number of cancer cells, and/or relieve to some extent one or more of the symptoms associated with the cancer. The effective amount may, for example, improve disease free survival (DFS), improve overall survival (OS), or decrease likelihood of recurrence.

Various chemical substituents are defined below. In some instances, the number of carbon atoms in a substituent (e.g., alkyl, alkanyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl, and aryl) is indicated by the prefix “C x -C y ” or “C x-y ” wherein x is the minimum and y is the maximum number of carbon atoms. Thus, for example, “C 1 -C 6 alkyl” refers to an alkyl containing from 1 to 6 carbon atoms. Illustrating further, “C 3 -C 8 cycloalkyl” means a saturated hydrocarbyl ring containing from 3 to 8 carbon ring atoms. If a substituent is described as being “substituted,” a hydrogen atom on a carbon or nitrogen is replaced with a non-hydrogen group. For example, a substituted alkyl substituent is an alkyl substituent in which at least one hydrogen atom on the alkyl is replaced with a non-hydrogen group. To illustrate, monofluoroalkyl is alkyl substituted with a fluoro radical, and difluoroalkyl is alkyl substituted with two fluoro radicals. It should be recognized that if there is more than one substitution on a substituent, each substitution may be identical or different (unless otherwise stated). If a substituent is described as being “optionally substituted”, the substituent may be either (1) not substituted or (2) substituted. Possible substituents include, but are not limited to, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, aryl, cycloalkyl, heterocyclyl, heteroaryl, halogen, C 1 -C 6 haloalkyl, oxo, —CN, NO 2 , —OR xa , —OC(O)R xz , —OC(O)N(R xa ) 2 , —SR xa , —S(O) 2 R xa , —S(O) 2 N(R xa ) 2 , —C(O)R xa , —C(O)OR xa , —C(O)N(R xa ) 2 , —C(O)N(R xa )S(O) 2 R xz , —N(R xa ) 2 , —N(R xa )C(O)R xz , —N(R xa )S(O) 2 R xz , —N(R xa )C(O)O(R xz ), —N(R xa )C(O)N(R xa ) 2 , —N(R xa )S(O) 2 N(R xa ) 2 , —(C 1 -C 6 alkylenyl)-CN, —(C 1 -C 6 alkylenyl)-OR xa , —(C 1 -C 6 alkylenyl)-OC(O)R xz , —(C 1- C 6 alkylenyl)-OC(O)N(R xa ) 2 , —(C 1 -C 6 alkylenyl)-SR xa , —(C 1 -C 6 alkylenyl)-S(O) 2 R xa , —(C 1- C 6 alkylenyl)-S(O) 2 N(R a ) 2 , —(C 1 -C 6 alkylenyl)-C(O)R xa , —(C 1 -C 6 alkylenyl)-C(O)OR xa , —(C 1 -C 6 alkylenyl)-C(O)N(R xa ) 2 , —(C 1 -C 6 alkylenyl)-C(O)N(R xa )S(O) 2 R xz , —(C 1 -C 6 alkylenyl)-N(R a ) 2 , —(C 1 -C 6 alkylenyl)-N(R xa )C(O)R z , —(C 1 -C 6 alkylenyl)-N(R xa )S(O) 2 R xz , —(C 1 -C 6 alkylenyl)-N(R xa )C(O)O(R z ), —(C 1 -C 6 alkylenyl)-N(R xa )C(O)N(R xa ) 2 , or —(C 1 -C 6 alkylenyl)-N(R xa )S(O) 2 N(R xa ) 2 ; wherein R xa , at each occurrence, is independently hydrogen, aryl, cycloalkyl, heterocyclyl, heteroaryl, C 1 -C 6 alkyl, or C 1 -C 6 haloalkyl; and R xz , at each occurrence, is independently aryl, cycloalkyl, heterocyclyl, heteroaryl, C 1 -C 6 alkyl or C 1 -C 6 haloalkyl.

›DETAILED DESCRIPTION OF THE INVENTION · 8 of 40

Various ADCs, synthons and Bcl-xL inhibitors comprising the ADCs and/or synthons are described in some embodiments herein by reference to structural formulae including substituents. It is to be understood that the various groups comprising substituents may be combined as valence and stability permit. Combinations of substituents and variables envisioned by this disclosure are only those that result in the formation of stable compounds. As used herein, the term “stable” refers to compounds that possess stability sufficient to allow manufacture and that maintain the integrity of the compound for a sufficient period of time to be useful for the purpose detailed herein.

As used herein, the following terms are intended to have the following meanings:

The term “alkoxy” refers to a group of the formula —OR xa , where R xa is an alkyl group. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy and the like.

The term “alkoxyalkyl” refers to an alkyl group substituted with an alkoxy group and may be represented by the general formula —R b OR xa where R b is an alkylene group and R xa is an alkyl group.

The term “alkyl” by itself or as part of another substituent refers to a saturated or unsaturated branched, straight-chain or cyclic monovalent hydrocarbon radical that is derived by the removal of one hydrogen atom from a single carbon atom of a parent alkane, alkene or alkyne. Typical alkyl groups include, but are not limited to, methyl; ethyls such as ethanyl, ethenyl, ethynyl; propyls such as propan-1-yl, propan-2-yl, cyclopropan-1-yl, prop-1-en-1-yl, prop-1-en-2-yl, prop-2-en-1-yl, cycloprop-1-en-1-yl; cycloprop-2-en-1-yl, prop-1-yn-1-yl, prop-2-yn-1-yl, etc.; butyls such as butan-1-yl, butan-2-yl, 2-methyl-propan-1-yl, 2-methyl-propan-2-yl, cyclobutan-1-yl, but-1-en-1-yl, but-1-en-2-yl, 2-methyl-prop-1-en-1-yl, but-2-en-1-yl, but-2-en-2-yl, buta-1,3-dien-1-yl, buta-1,3-dien-2-yl, cyclobut-1-en-1-yl, cyclobut-1-en-3-yl, cyclobuta-1,3-dien-1-yl, but-1-yn-1-yl, but-1-yn-3-yl, but-3-yn-1-yl, etc.; and the like. Where specific levels of saturation are intended, the nomenclature “alkanyl,” “alkenyl” and/or “alkynyl” are used, as defined below. The term “lower alkyl” refers to alkyl groups with 1 to 6 carbons.

The term “alkanyl” by itself or as part of another substituent refers to a saturated branched, straight-chain or cyclic alkyl derived by the removal of one hydrogen atom from a single carbon atom of a parent alkane. Typical alkanyl groups include, but are not limited to, methyl; ethanyl; propanyls such as propan-1-yl, propan-2-yl (isopropyl), cyclopropan-1-yl, etc.; butanyls such as butan-1-yl, butan-2-yl (sec-butyl), 2-methyl-propan-1-yl (isobutyl), 2-methyl-propan-2-yl (t-butyl), cyclobutan-1-yl, etc.; and the like.

The term “alkenyl” by itself or as part of another substituent refers to an unsaturated branched, straight-chain or cyclic alkyl having at least one carbon-carbon double bond derived by the removal of one hydrogen atom from a single carbon atom of a parent alkene. Typical alkenyl groups include, but are not limited to, ethenyl; propenyls such as prop-1-en-1-yl, prop-1-en-2-yl, prop-2-en-1-yl, prop-2-en-2-yl, cycloprop-1-en-1-yl; cycloprop-2-en-1-yl; butenyls such as but-1-en-1-yl, but-1-en-2-yl, 2-methyl-prop-1-en-1-yl, but-2-en-1-yl, but-2-en-2-yl, buta-1,3-dien-1-yl, buta-1,3-dien-2-yl, cyclobut-1-en-1-yl, cyclobut-1-en-3-yl, cyclobuta-1,3-dien-1-yl, etc.; and the like.

The term “alkynyl” by itself or as part of another substituent refers to an unsaturated branched, straight-chain or cyclic alkyl having at least one carbon-carbon triple bond derived by the removal of one hydrogen atom from a single carbon atom of a parent alkyne. Typical alkynyl groups include, but are not limited to, ethynyl; propynyls such as prop-1-yn-1-yl, prop-2-yn-1-yl, etc.; butynyls such as but-1-yn-1-yl, but-1-yn-3-yl, but-3-yn-1-yl, etc.; and the like.

The term “alkylamine” refers to a group of the formula —NHR xa and “dialkylamine” refers to a group of the formula —NR a R xa , where each R xa is, independently of the others, an alkyl group.

The term “alkylene” refers to an alkane, alkene or alkyne group having two terminal monovalent radical centers derived by the removal of one hydrogen atom from each of the two terminal carbon atoms. Typical alkylene groups include, but are not limited to, methylene; and saturated or unsaturated ethylene; propylene; butylene; and the like. The term “lower alkylene” refers to alkylene groups with 1 to 6 carbons.

The term “heteroalkylene” refers to a divalent alkylene having one or more —CH 2 — groups replaced with a thio, oxy, or —NR x3 — where R x3 is selected from hydrogen, lower alkyl and lower heteroalkyl. The heteroalkylene can be linear, branched, cyclic, bicyclic, or a combination thereof and can include up to 10 carbon atoms and up to 4 heteroatoms. The term “lower heteroalkylene” refers to alkylene groups with 1 to 4 carbon atoms and 1 to 3 heteroatoms.

The term “aryl” means an aromatic carbocyclyl containing from 6 to 14 carbon ring atoms. An aryl may be monocyclic or polycyclic (i.e., may contain more than one ring). In the case of polycyclic aromatic rings, only one ring the polycyclic system is required to be aromatic while the remaining ring(s) may be saturated, partially saturated or unsaturated. Examples of aryls include phenyl, naphthalenyl, indenyl, indanyl, and tetrahydronaphthyl.

The term “arylene” refers to an aryl group having two monovalent radical centers derived by the removal of one hydrogen atom from each of the two ring carbons. An exemplary arylene group is a phenylene.

An alkyl group may be substituted by a “carbonyl” which means that two hydrogen atoms from a single alkanylene carbon atom are removed and replaced with a double bond to an oxygen atom.

The prefix “halo” indicates that the substituent which includes the prefix is substituted with one or more independently selected halogen radicals. For example, haloalkyl means an alkyl substituent in which at least one hydrogen radical is replaced with a halogen radical. Typical halogen radicals include chloro, fluoro, bromo and iodo. Examples of haloalkyls include chloromethyl, 1-bromoethyl, fluoromethyl, difluoromethyl, trifluoromethyl, and 1,1,1-trifluoroethyl. It should be recognized that if a substituent is substituted by more than one halogen radical, those halogen radicals may be identical or different (unless otherwise stated).

›DETAILED DESCRIPTION OF THE INVENTION · 9 of 40

The term “haloalkoxy” refers to a group of the formula —OR c , where R c is a haloalkyl.

The terms “heteroalkyl,” “heteroalkanyl,” “heteroalkenyl,” “heteroalkynyl,” and “heteroalkylene” refer to alkyl, alkanyl, alkenyl, alkynyl, and alkylene groups, respectively, in which one or more of the carbon atoms, e.g., 1, 2 or 3 carbon atoms, are each independently replaced with the same or different heteroatoms or heteroatomic groups. Typical heteroatoms and/or heteroatomic groups which can replace the carbon atoms include, but are not limited to, —O—, —S—, —S—O—, —NR c —, —PH, —S(O)—, —S(O) 2 —, —S(O)NR c —, —S(O) 2 NR c —, and the like, including combinations thereof, where each R c is independently hydrogen or C 1 -C 6 alkyl. The term “lower heteroalkyl” refers to between 1 and 4 carbon atoms and between 1 and 3 heteroatoms.

The terms “cycloalkyl” and “heterocyclyl” refer to cyclic versions of “alkyl” and “heteroalkyl” groups, respectively. For heterocyclyl groups, a heteroatom can occupy the position that is attached to the remainder of the molecule. A cycloalkyl or heterocyclyl ring may be a single-ring (monocyclic) or have two or more rings (bicyclic or polycyclic).

Monocyclic cycloalkyl and heterocyclyl groups will typically contains from 3 to 7 ring atoms, more typically from 3 to 6 ring atoms, and even more typically 5 to 6 ring atoms. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl; cyclobutyls such as cyclobutanyl and cyclobutenyl; cyclopentyls such as cyclopentanyl and cyclopentenyl; cyclohexyls such as cyclohexanyl and cyclohexenyl; and the like. Examples of monocyclic heterocyclyls include, but are not limited to, oxetane, furanyl, dihydrofuranyl, tetrahydrofuranyl, tetrahydropyranyl, thiophenyl (thiofuranyl), dihydrothiophenyl, tetrahydrothiophenyl, pyrrolyl, pyrrolinyl, pyrrolidinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyrazolyl, pyrazolinyl, pyrazolidinyl, triazolyl, tetrazolyl, oxazolyl, oxazolidinyl, isoxazolidinyl, isoxazolyl, thiazolyl, isothiazolyl, thiazolinyl, isothiazolinyl, thiazolidinyl, isothiazolidinyl, thiodiazolyl, oxadiazolyl (including 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl (furazanyl), or 1,3,4-oxadiazolyl), oxatriazolyl (including 1,2,3,4-oxatriazolyl or 1,2,3,5-oxatriazolyl), dioxazolyl (including 1,2,3-dioxazolyl, 1,2,4-dioxazolyl, 1,3,2-dioxazolyl, or 1,3,4-dioxazolyl), 1,4-dioxanyl, dioxothiomorpholinyl, oxathiazolyl, oxathiolyl, oxathiolanyl, pyranyl, dihydropyranyl, thiopyranyl, tetrahydrothiopyranyl, pyridinyl (azinyl), piperidinyl, diazinyl (including pyridazinyl (1,2-diazinyl), pyrimidinyl (1,3-diazinyl), or pyrazinyl (1,4-diazinyl)), piperazinyl, triazinyl (including 1,3,5-triazinyl, 1,2,4-triazinyl, and 1,2,3-triazinyl)), oxazinyl (including 1,2-oxazinyl, 1,3-oxazinyl, or 1,4-oxazinyl)), oxathiazinyl (including 1,2,3-oxathiazinyl, 1,2,4-oxathiazinyl, 1,2,5-oxathiazinyl, or 1,2,6-oxathiazinyl)), oxadiazinyl (including 1,2,3-oxadiazinyl, 1,2,4-oxadiazinyl, 1,4,2-oxadiazinyl, or 1,3,5-oxadiazinyl)), morpholinyl, azepinyl, oxepinyl, thiepinyl, diazepinyl, pyridonyl (including pyrid-2(1H)-onyl and pyrid-4(1H)-onyl), furan-2(5H)-onyl, pyrimidonyl (including pyramid-2(1H)-onyl and pyramid-4(3H)-onyl), oxazol-2(3H)-onyl, 1H-imidazol-2(3H)-onyl, pyridazin-3(2H)-onyl, and pyrazin-2(1H)-onyl.

Polycyclic cycloalkyl and heterocyclyl groups contain more than one ring, and bicyclic cycloalkyl and heterocyclyl groups contain two rings. The rings may be in a bridged, fused or spiro orientation. Polycyclic cycloalkyl and heterocyclyl groups may include combinations of bridged, fused and/or spiro rings. In a spirocyclic cycloalkyl or heterocyclyl, one atom is common to two different rings. An example of a spirocycloalkyl is spiro[4.5]decane and an example of a spiroheterocyclyls is a spiropyrazoline.

In a bridged cycloalkyl or heterocyclyl, the rings share at least two common non-adjacent atoms. Examples of bridged cycloalkyls include, but are not limited to, adamantyl and norbornanyl rings. Examples of bridged heterocyclyls include, but are not limited to, 2-oxatricyclo[3.3.1.1 3,7 ]decanyl.

In a fused-ring cycloalkyl or heterocyclyl, two or more rings are fused together, such that two rings share one common bond. Examples of fused-ring cycloalkyls include decalin, naphthylene, tetralin, and anthracene. Examples of fused-ring heterocyclyls containing two or three rings include imidazopyrazinyl (including imidazo[1,2-a]pyrazinyl), imidazopyridinyl (including imidazo[1,2-a]pyridinyl), imidazopyridazinyl (including imidazo[1,2-b]pyridazinyl), thiazolopyridinyl (including thiazolo[5,4-c]pyridinyl, thiazolo[5,4-b]pyridinyl, thiazolo[4,5-b]pyridinyl, and thiazolo[4,5-c]pyridinyl), indolizinyl, pyranopyrrolyl, 4H-quinolizinyl, purinyl, naphthyridinyl, pyridopyridinyl (including pyrido[3,4-b]-pyridinyl, pyrido[3,2-b]-pyridinyl, or pyrido[4,3-b]-pyridinyl), and pteridinyl. Other examples of fused-ring heterocyclyls include benzo-fused heterocyclyls, such as dihydrochromenyl, tetrahydroisoquinolinyl, indolyl, isoindolyl (isobenzazolyl, pseudoisoindolyl), indoleninyl (pseudoindolyl), isoindazolyl (benzpyrazolyl), benzazinyl (including quinolinyl (1-benzazinyl) or isoquinolinyl (2-benzazinyl)), phthalazinyl, quinoxalinyl, quinazolinyl, benzodiazinyl (including cinnolinyl (1,2-benzodiazinyl) or quinazolinyl (1,3-benzodiazinyl)), benzopyranyl (including chromanyl or isochromanyl), benzoxazinyl (including 1,3,2-benzoxazinyl, 1,4,2-benzoxazinyl, 2,3,1-benzoxazinyl, or 3,1,4-benzoxazinyl), benzo[d]thiazolyl, and benzisoxazinyl (including 1,2-benzisoxazinyl or 1,4-benzisoxazinyl).

The term “cycloalkylene” refers to a cycloalkyl group having two monovalent radical centers derived by the removal of one hydrogen atom from each of two ring carbons. Exemplary cycloalkylene groups include:

The term “heteroaryl” refers to an aromatic heterocyclyl containing from 5 to 14 ring atoms. A heteroaryl may be a single ring or 2 or 3 fused rings. Examples of heteroaryls include 6-membered rings such as pyridyl, pyrazyl, pyrimidinyl, pyridazinyl, and 1,3,5-, 1,2,4- or 1,2,3-triazinyl; 5-membered ring substituents such as triazolyl, pyrrolyl, imidazyl, furanyl, thiophenyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, 1,2,3-, 1,2,4-, 1,2,5-, or 1,3,4-oxadiazolyl and isothiazolyl; 6/5-membered fused ring substituents such as imidazopyrazinyl (including imidazo[1,2-a]pyrazinyl)imidazopyridinyl (including imidazo[1,2-a]pyridinyl), imidazopyridazinyl (including imidazo[1,2-b]pyridazinyl), thiazolopyridinyl (including thiazolo[5,4-c]pyridinyl, thiazolo[5,4-b]pyridinyl, thiazolo[4,5-b]pyridinyl, and thiazolo[4,5-c]pyridinyl), benzo[d]thiazolyl, benzothiofuranyl, benzisoxazolyl, benzoxazolyl, purinyl, and anthranilyl; and 6/6-membered fused rings such as benzopyranyl, quinolinyl, isoquinolinyl, cinnolinyl, quinazolinyl, and benzoxazinyl. Heteroaryls may also be heterocycles having aromatic (4N+2 pi electron) resonance contributors such as pyridonyl (including pyrid-2(1H)-onyl and pyrid-4(1H)-onyl), pyrimidonyl (including pyramid-2(1H)-onyl and pyramid-4(3H)-onyl), pyridazin-3(2H)-onyl and pyrazin-2(1H)-onyl.

›DETAILED DESCRIPTION OF THE INVENTION · 10 of 40

The term “sulfonate” as used herein means a salt or ester of a sulfonic acid.

The term “methyl sulfonate” as used herein means a methyl ester of a sulfonic acid group.

The term “carboxylate” as used herein means a salt or ester of a carboxylic acid.

The term “polyol”, as used herein, means a group containing more than two hydroxyl groups independently or as a portion of a monomer unit. Polyols include, but are not limited to, reduced C 2 -C 6 carbohydrates, ethylene glycol, and glycerin.

The term “sugar” when used in context of “G 1 ” includes O-glycoside, N-glycoside, S-glycoside and C-glycoside (C-glycoslyl) carbohydrate derivatives of the monosaccharide and disaccharide classes and may originate from naturally-occurring sources or may be synthetic in origin.

For example “sugar” when used in context of “G 1 ” includes derivatives such as but not limited to those derived from glucuronic acid, galacturonic acid, galactose, and glucose among others. Suitable sugar substitutions include but are not limited to hydroxyl, amine, carboxylic acid, sulfonic acid, phosphonic acid, esters, and ethers.

The term “NHS ester” means the N-hydroxysuccinimide ester derivative of a carboxylic acid.

The term “amine” includes primary, secondary and tertiary aliphatic amines, including cyclic versions.

The term salt when used in context of “or salt thereof” include salts commonly used to form alkali metal salts and to form addition salts of free acids or free bases. In general, these salts typically may be prepared by conventional means by reacting, for example, the appropriate acid or base with a compound of the invention

Where a salt is intended to be administered to a patient (as opposed to, for example, being in use in an in vitro context), the salt preferably is pharmaceutically acceptable and/or physiologically compatible. The term “pharmaceutically acceptable” is used adjectivally in this patent application to mean that the modified noun is appropriate for use as a pharmaceutical product or as a part of a pharmaceutical product. The term “pharmaceutically acceptable salt” includes salts commonly used to form alkali metal salts and to form addition salts of free acids or free bases. In general, these salts typically may be prepared by conventional means by reacting, for example, the appropriate acid or base with a compound of the invention.

Various aspects of the invention are described in further detail in the following subsections.

II. Anti-B7-H3 Antibodies

One aspect of the invention provides anti-B7-H3 antibodies, or antigen binding portions thereof. In one embodiment, the present invention provides chimeric anti-B7-H3 antibodies, or antigen binding portions thereof. In yet another embodiment, the present invention provides humanized anti-B7-H3 antibodies, or antigen binding portions thereof. In another aspect, the invention features antibody drug conjugates (ADCs) comprising an anti-B7-H3 antibody described herein and at least one drug(s), such as, but not limited to, a Bcl-xL inhibitor or an auristatin. The antibodies or ADCs of the invention have characteristics including, but not limited to, binding to wild-type human B7-H3 in vitro, binding to wild-type human B7-H3 on tumor cells expressing B7-H3, and decreasing or inhibiting xenograft tumor growth in a mouse model.

One aspect of the invention features an anti-human B7-H3 (anti-hB7-H3) Antibody Drug Conjugate (ADC) comprising an anti-hB7-H3 antibody conjugated to a drug via a linker, wherein the drug is a Bcl-xL inhibitor. Exemplary anti-B7-H3 antibodies (and sequences thereof) that can be used in the ADCs are described herein.

The anti-B7-H3 antibodies described herein provide the ADCs of the invention with the ability to bind to B7-H3 such that the cytotoxic Bcl-xL drug attached to the antibody may be delivered to the B7-H3-expressing cell, particularly a B7-H3 expressing cancer cell.

While the term “antibody” is used throughout, it should be noted that antibody fragments (i.e., antigen-binding portions of an anti-B7-H3antibody) are also included in the invention and may be included in the embodiments (methods and compositions) described throughout. For example, an anti-B7-H3antibody fragment may be conjugated to the Bcl-xL inhibitors described herein. Thus, it is within the scope of the invention that in certain embodiments, antibody fragments of the anti-B7-H3antibodies described herein are conjugated to Bcl-xL inhibitors (including those described below in Section III.A) via linkers (including those described below in Section III.A). In certain embodiments, the anti-B7-H3 antibody binding portion is a Fab, a Fab′, a F(ab′)2, a Fv, a disulfide linked Fv, an scFv, a single domain antibody, or a diabody.

II.A. Anti-B7-H3 Chimeric Antibodies

A chimeric antibody is a molecule in which different portions of the antibody are derived from different animal species, such as antibodies having a variable region derived from a murine monoclonal antibody and a human immunoglobulin constant region. Methods for producing chimeric antibodies are known in the art. See e.g., Morrison, Science 229:1202 (1985); Oi et al., BioTechniques 4:214 (1986); Gillies et al., (1989) J. Immunol. Methods 125:191-202; U.S. Pat. Nos. 5,807,715; 4,816,567; and 4,816,397, which are incorporated herein by reference in their entireties. In addition, techniques developed for the production of “chimeric antibodies” (Morrison et al., 1984 , Proc. Natl. Acad. Sci. 81:851-855; Neuberger et al., 1984 , Nature 312:604-608; Takeda et al., 1985 , Nature 314:452-454, each of which are incorporated herein by reference in their entireties) by splicing genes from a mouse antibody molecule of appropriate antigen specificity together with genes from a human antibody molecule of appropriate biological activity can be used.

As described in Example 3, eighteen anti-B7-H3 murine antibodies were identified having high specific binding activity against human and cynomolgus B7-H3. Chimeric antibodies, in the context of a human immunoglobulin constant region, were generated from these eighteen antibodies.

›DETAILED DESCRIPTION OF THE INVENTION · 11 of 40

Thus, in one aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable region including an amino acid sequence set forth in SEQ ID NOs: 1, 9, 16, 24, 32, 40, 48, 56, 64, 72, 80, 87, 95, 101, or 108; and/or a light chain variable region including an amino acid sequence set forth in SEQ ID NOs: 5, 13, 20, 28, 36, 44, 52, 60, 68, 76, 84, 91, 98, 105, or 112.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable region including an amino acid sequence as set forth in SEQ ID NO: 1, and a light chain variable region including an amino acid sequence set forth in SEQ ID NO: 5.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 2; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 3; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 4; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 6; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 7; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 8.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable region including an amino acid sequence as set forth in SEQ ID NO: 9, and a light chain variable region including an amino acid sequence set forth in SEQ ID NO: 13.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 10; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 11; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 12; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 14 (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 7; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 15.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable region including an amino acid sequence as set forth in SEQ ID NO: 16, and a light chain variable region including an amino acid sequence set forth in SEQ ID NO: 20.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 17; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 18; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 19; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 21; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 22; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 23.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable region including an amino acid sequence as set forth in SEQ ID NO: 24, and a light chain variable region including an amino acid sequence set forth in SEQ ID NO: 28.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 25; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 26; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 27; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 29; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 30; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 31.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable region including an amino acid sequence as set forth in SEQ ID NO: 32, and a light chain variable region including an amino acid sequence set forth in SEQ ID NO: 36.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 33; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 34; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 35; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 37; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 38; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 182.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable region including an amino acid sequence as set forth in SEQ ID NO: 40, and a light chain variable region including an amino acid sequence set forth in SEQ ID NO: 44.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 41; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 42; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 43; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 45; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 46; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 47.

›DETAILED DESCRIPTION OF THE INVENTION · 12 of 40

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable region including an amino acid sequence as set forth in SEQ ID NO: 48, and a light chain variable region including an amino acid sequence set forth in SEQ ID NO: 52.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 49; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 50; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 51; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 53; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 54; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 55.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable region including an amino acid sequence as set forth in SEQ ID NO: 56, and a light chain variable region including an amino acid sequence set forth in SEQ ID NO: 60.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 57; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 58; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 59; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 61; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 62; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 63.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable region including an amino acid sequence as set forth in SEQ ID NO: 64, and a light chain variable region including an amino acid sequence set forth in SEQ ID NO: 68.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 65; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 66; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 67; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 69; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 70; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 71.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable region including an amino acid sequence as set forth in SEQ ID NO: 72, and a light chain variable region including an amino acid sequence set forth in SEQ ID NO: 76.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 73; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 74; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 75; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 77; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 78; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 79.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable region including an amino acid sequence as set forth in SEQ ID NO: 80, and a light chain variable region including an amino acid sequence set forth in SEQ ID NO: 84.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 81; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 82; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 83; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 85; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 7; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 86.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable region including an amino acid sequence as set forth in SEQ ID NO: 87, and a light chain variable region including an amino acid sequence set forth in SEQ ID NO: 91.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 88; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 89; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 90; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 92; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 93; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 94.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable region including an amino acid sequence as set forth in SEQ ID NO: 95, and a light chain variable region including an amino acid sequence set forth in SEQ ID NO: 98.

›DETAILED DESCRIPTION OF THE INVENTION · 13 of 40

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 49; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 96; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 97; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 99; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 93; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 100.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable region including an amino acid sequence as set forth in SEQ ID NO: 101, and a light chain variable region including an amino acid sequence set forth in SEQ ID NO: 105.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 102; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 103; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 104; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 106; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 46; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 107.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable region including an amino acid sequence as set forth in SEQ ID NO: 108, and a light chain variable region including an amino acid sequence set forth in SEQ ID NO: 112.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 109; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 110; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 111; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 113; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 114; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 115.

II.B. Humanized Anti-B7-H3 Antibodies

The chimeric antibodies disclosed herein may be used in the production of humanized anti-B7-H3 antibodies. For example, following the generation and characterization of chimeric anti-B7-H3 antibodies chAb1-chAb18, antibodies chAb3, chAb13, and chAb18 were selected for humanization. Specifically, six different humanized antibodies were created based on chAb3 (referred to herein as huAb3v1, huAb3v2, huAb3v3, huAb3v4, huAb3v5, and huAb3v6 (see Examples 12 and 13), nine different humanized antibodies were created based on chAb13 (referred to herein as huAb13v1, huAb13v2, huAb13v3, huAb13v4, huAb13v5, huAb13v6, huAb13v7, huAb13v8, huAb13v9), and ten different humanized antibodies were created based on chAb18 (referred to herein as huAb18v1, huAb18v2, huAb18v3, huAb18v4, huAb18v5, huAb18v6, huAb18v7, huAb18v8, huAb18v9, and huAb18v10 (see Examples 9 and 10)). Tables 8, 12, 16, 18, and 19 provide the amino acid sequences of CDR, VH and VL regions of humanized chAb3, chAb13, and chAb18, respectively.

Generally, humanized antibodies are antibody molecules from non-human species antibody that binds the desired antigen having one or more complementarity determining regions (CDRs) from the non-human species and framework regions from a human immunoglobulin molecule. Known human Ig sequences are disclosed, e.g., www.ncbi.nlm.nih.gov/entrez-/query.fcgi;

www.atcc.org/phage/hdb.html; www.sciquest.com/; www.abcam.com/; www.antibodyresource.com/onlinecomp.html; www.public.iastate.edu/.about.pedro/research_tools.html; www.mgen.uni-heidelberg.de/SD/IT/IT.html; www.whfreeman.com/immunology/CH-05/kuby05.htm; www.library.thinkquest.org/12429/Immune/Antibody.html; www.hhmi.org/grants/lectures/1996/vlab/; www.path.cam.ac.uk/.about.mrc7/m-ikeimages.html; www.antibodyresource.com/; mcb.harvard.edu/BioLinks/Immuno-logy.html.www.immunologylink.com/; pathbox.wustl.edu/.about.hcenter/index.-html; www.biotech.ufl.edu/.about.hcl/; www.pebio.com/pa/340913/340913.html-; www.nal.usda.gov/awic/pubs/antibody/; www.m.ehime-u.acjp/.about.yasuhito-/Elisa.html; www.biodesign.com/table.asp; www.icnet.uk/axp/facs/davies/lin-ks.html; www.biotech.ufl.edu/.about.fccl/protocol.html; www.isac-net.org/sites_geo.html; aximtl.imt.uni-marburg.de/.about.rek/AEP-Start.html; baserv.uci.kun.nl/.about.jraats/linksl.html; www.recab.uni-hd.de/immuno.bme.nwu.edu/; www.mrc-cpe.cam.ac.uk/imt-doc/pu-blic/INTRO.html; www.ibt.unam.mx/vir/V_mice.html; imgt.cnusc.fr:8104/; www.biochem.ucl.ac.uk/.about.martin/abs/index.html; antibody.bath.ac.uk/; abgen.cvm.tamu.edu/lab/wwwabgen.html; www.unizh.ch/.about.honegger/AHOsem-inar/Slide01.html; www.cryst.bbk.ac.uk/.about.ubcg07s/; www.nimr.mrc.ac.uk/CC/ccaewg/ccaewg.htm; www.path.cam.ac.uk/.about.mrc7/h-umanisation/TAHHP.html; www.ibt.unam.mx/vir/structure/stat_aim.html; www.biosci.missouri.edu/smithgp/index.html; www.cryst.bioc.cam.ac.uk/.abo-ut.fmolina/Web-pages/Pept/spottech.html; www.jerini.de/fr roducts.htm; www.patents.ibm.com/ibm.html.Kabat et al.,

Sequences of Proteins of Immunological Interest, U.S. Dept. Health (1983), each entirely incorporated herein by reference. Such imported sequences can be used to reduce immunogenicity or reduce, enhance or modify binding, affinity, on-rate, off-rate, avidity, specificity, half-life, or any other suitable characteristic, as known in the art.

Framework residues in the human framework regions may be substituted with the corresponding residue from the CDR donor antibody to alter, preferably improve, antigen binding. These framework substitutions are identified by methods well known in the art, e.g., by modeling of the interactions of the CDR and framework residues to identify framework residues important for antigen binding and sequence comparison to identify unusual framework residues at particular positions. (See, e.g., Queen et al., U.S. Pat. No. 5,585,089; Riechmann et al., Nature 332:323 (1988), which are incorporated herein by reference in their entireties.) Three-dimensional immunoglobulin models are commonly available and are familiar to those skilled in the art. Computer programs are available which illustrate and display probable three-dimensional conformational structures of selected candidate immunoglobulin sequences. Inspection of these displays permits analysis of the likely role of the residues in the functioning of the candidate immunoglobulin sequence, i.e., the analysis of residues that influence the ability of the candidate immunoglobulin to bind its antigen. In this way, FR residues can be selected and combined from the consensus and import sequences so that the desired antibody characteristic, such as increased affinity for the target antigen(s), is achieved. In general, the CDR residues are directly and most substantially involved in influencing antigen binding. Antibodies can be humanized using a variety of techniques known in the art, such as but not limited to those described in Jones et al., Nature 321:522 (1986); Verhoeyen et al., Science 239:1534 (1988)), Sims et al., J. Immunol. 151: 2296 (1993); Chothia and Lesk, J. Mol. Biol. 196:901 (1987), Carter et al., Proc. Natl. Acad. Sci. U.S.A. 89:4285 (1992); Presta et al., J. Immunol. 151:2623 (1993), Padlan, Molecular Immunology 28(4/5):489-498 (1991); Studnicka et al., Protein Engineering 7(6):805-814 (1994); Roguska. et al., PNAS 91:969-973 (1994); PCT publication WO 91/09967, PCT/: US98/16280, US96/18978, US91/09630, US91/05939, US94/01234, GB89/01334, GB91/01134, GB92/01755; WO90/14443, WO90/14424, WO90/14430, EP 229246, EP 592,106; EP 519,596, EP 239,400, U.S. Pat. Nos. 5,565,332, 5,723,323, 5,976,862, 5,824,514, 5,817,483, 5,814,476, 5,763,192, 5,723,323, 5,766886, 5,714,352, 6,204,023, 6,180,370, 5,693,762, 5,530,101, 5,585,089, 5,225,539; 4,816,567, each entirely incorporated herein by reference, included references cited therein.

›DETAILED DESCRIPTION OF THE INVENTION · 14 of 40

Humanized Anti-B7-H3 Antibodies Derived from chAb3

Six humanized antibodies based on chAb3 were created. The sequences of each are as follows:

A) huAb3v1 (VH amino acid sequence set forth in SEQ ID NO: 125 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 10, 11, and 12, respectively; and VL amino acid sequence set forth in SEQ ID NO: 128 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 14, 7, and 15, respectively);

B) huAb3v2 (VH amino acid sequence set forth in SEQ ID NO: 127 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 10, 11, and 12, respectively; and VL amino acid sequence set forth in SEQ ID NO: 128 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 14, 7, and 15, respectively);

C) huAb3v3 (VH amino acid sequence set forth in SEQ ID NO: 126 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 10, 11, and 12, respectively; and VL amino acid sequence set forth in SEQ ID NO: 129 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 14, 7, and 15, respectively);

D) huAb3v4 (VH amino acid sequence set forth in SEQ ID NO: 125 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 10, 11, and 12, respectively; and VL amino acid sequence set forth in SEQ ID NO: 130 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 14, 7, and 15, respectively);

E) huAb3v5 (VH amino acid sequence set forth in SEQ ID NO: 127 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 10, 11, and 12, respectively; and VL amino acid sequence set forth in SEQ ID NO: 130 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 14, 7, and 15, respectively); and

F) huAb3v6 (VH amino acid sequence set forth in SEQ ID NO: 126 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 10, 11, and 12, respectively; and VL amino acid sequence set forth in SEQ ID NO: 130 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 14, 7, and 15, respectively).

Of the six humanized versions of chAb3, huAb3v2 was selected for further modified in order to remove potential deamidation or isomerization sites in the light chain CDR1 or in the heavy chain CDR2. Nine variants of the humanized antibody huAb3v2 were generated, and are referred to herein as huAb3v2.1, huAb3v2.2, huAb3v2.3, huAb3v2.4, huAb3v2.5, huAb3v2.6, huAb3v2.7, huAb3v2.8, and huAb3v2.9 (CDR and variable domain sequences are provided in Table 16). The nine variants of the huAb3v2 antibody include the following:

A) huAb3v2.1 (VH amino acid sequence set forth in SEQ ID NO: 131 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 10, 132, and 12, respectively; and VL amino acid sequence set forth in SEQ ID NO: 133 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 134, 7, and 15, respectively);

B) huAb3v2.2 (VH amino acid sequence set forth in SEQ ID NO: 131 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 10, 132, and 12, respectively; and VL amino acid sequence set forth in SEQ ID NO: 135 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 136, 7, and 15, respectively);

C) huAb3v2.3 (VH amino acid sequence set forth in SEQ ID NO: 131 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 10, 132, and 12, respectively; and VL amino acid sequence set forth in SEQ ID NO: 137 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 138, 7, and 15, respectively);

D) huAb3v2.4 (VH amino acid sequence set forth in SEQ ID NO: 139 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 10, 140, and 12, respectively; and VL amino acid sequence set forth in SEQ ID NO: 133 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 134, 7, and 15, respectively);

E) huAb3v2.5 (VH amino acid sequence set forth in SEQ ID NO: 139 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 10, 140, and 12, respectively; and VL amino acid sequence set forth in SEQ ID NO: 135 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 136, 7, and 15, respectively);

F) huAb3v2.6 (VH amino acid sequence set forth in SEQ ID NO: 139 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 10, 140, and 12, respectively; and VL amino acid sequence set forth in SEQ ID NO: 137 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 138, 7, and 15, respectively);

G) huAb3v2.7 (VH amino acid sequence set forth in SEQ ID NO: 141 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 10, 142, and 12, respectively; and VL amino acid sequence set forth in SEQ ID NO: 133 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 134, 7, and 15, respectively);

H) huAb3v2.8 (VH amino acid sequence set forth in SEQ ID NO: 141 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 10, 142, and 12, respectively; and VL amino acid sequence set forth in SEQ ID NO: 135 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 136, 7, and 15, respectively); and

I) huAb3v2.9 (VH amino acid sequence set forth in SEQ ID NO: 141 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 10, 142, and 12, respectively; and VL amino acid sequence set forth in SEQ ID NO: 137 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 138, 7, and 15, respectively).

Thus, in one aspect, the present invention provides antibodies comprising variable and/or CDR sequences from a humanized antibody derived from chAb3. In one embodiment, the invention features anti-B7-H3 antibodies which are derived from Ab3 have improved characteristics, e.g., improved binding affinity to isolated B7-H3 protein and improved binding to B7-H3 expressing cells, as described in the Examples below. Collectively these novel antibodies are referred to herein as “Ab3 variant antibodies.” Generally, the Ab3 variant antibodies retain the same epitope specificity as Ab3. In various embodiments, anti-B7-H3 antibodies, or antigen binding fragments thereof, of the invention are capable of modulating a biological function of B7-H3.

›DETAILED DESCRIPTION OF THE INVENTION · 15 of 40

In one aspect, the present invention provides a humanized antibody, or antigen binding portion thereof, having a heavy chain variable region including an amino acid sequence set forth in SEQ ID NOs: 125, 126, 127, 131, 139, or 141; and/or a light chain variable region including an amino acid sequence set forth in SEQ ID NOs: 128, 129, 130, 133, 135, or 137.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen binding portion thereof, of the invention comprises a heavy chain variable region comprising a CDR1 domain comprising an amino acid sequence as set forth in SEQ ID NO: 10; a CDR2 domain comprising an amino acid sequence as set forth in SEQ ID NO: 11, 132, 140, or 142; and a CDR3 domain comprising an amino acid sequence as set forth in SEQ ID NO: 12; and a light chain variable region comprising a CDR1 domain comprising an amino acid sequence as set forth in SEQ ID NO: 14, 134, 136, or 138; a CDR2 domain comprising an amino acid sequence as set forth in SEQ ID NO: 7; and a CDR3 domain comprising an amino acid sequence as set forth in SEQ ID NO: 15.

In another aspect, the present invention is directed to a humanized anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 10; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 11; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 12; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 14; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 7; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 15.

In another aspect, the present invention is directed to a humanized anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 10; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 132; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 12; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 134; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 7; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 15.

In another aspect, the present invention is directed to a humanized anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 10; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 132; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 12; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 136; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 7; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 15.

In another aspect, the present invention is directed to a humanized anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 10; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 132; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 12; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 138; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 7; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 15.

›DETAILED DESCRIPTION OF THE INVENTION · 16 of 40

In another aspect, the present invention is directed to a humanized anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 10; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 140; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 12; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 134; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 7; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 15.

In another aspect, the present invention is directed to a humanized anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 10; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 140; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 12; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 136; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 7; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 15.

In another aspect, the present invention is directed to a humanized anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 10; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 140; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 12; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 138; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 7; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 15.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain comprising the amino acid sequence of SEQ ID NO: 172 and a light chain comprising the amino acid sequence of SEQ ID NO: 173. In one aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable region including an amino acid sequence as set forth in SEQ ID NO: 141, and a light chain variable region including an amino acid sequence set forth in SEQ ID NO: 133.

In another aspect, the present invention is directed to a humanized anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 10; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 142; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 12; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 134; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 7; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 15.

In another aspect, the present invention is directed to a humanized anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 10; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 142; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 12; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 136; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 7; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 15.

In another aspect, the present invention is directed to a humanized anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 10; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 142; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 12; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 138; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 7; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 15.

›DETAILED DESCRIPTION OF THE INVENTION · 17 of 40

Humanized Anti-B7-H3 Antibodies Derived from chAb13

The nine different humanized antibodies created based on chAb13 include the following:

A) huAb13v1 (VH amino acid sequence set forth in SEQ ID NO: 147 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 33, 34, and 35, respectively; and VL amino acid sequence set forth in SEQ ID NO: 144 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 37, 38, and 39, respectively);

B) huAb13v2 (VH amino acid sequence set forth in SEQ ID NO: 146 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 33, 34, and 35, respectively; and VL amino acid sequence set forth in SEQ ID NO: 143 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 37, 38, and 39, respectively);

C) huAb13v3 (VH amino acid sequence set forth in SEQ ID NO: 146 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 33, 34, and 35, respectively; and VL amino acid sequence set forth in SEQ ID NO: 144 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 37, 38, and 39, respectively);

D) huAb13v4 (VH amino acid sequence set forth in SEQ ID NO: 146 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 33, 34, and 35, respectively; and VL amino acid sequence set forth in SEQ ID NO: 145 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 37, 38, and 39, respectively);

E) huAb13v5 (VH amino acid sequence set forth in SEQ ID NO: 147 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 33, 34, and 35, respectively; and VL amino acid sequence set forth in SEQ ID NO: 143 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 37, 38, and 39, respectively);

F) huAb13v6 (VH amino acid sequence set forth in SEQ ID NO: 147 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 33, 34, and 35, respectively; and VL amino acid sequence set forth in SEQ ID NO: 145 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 37, 38, and 39, respectively);

G) huAb13v7 (VH amino acid sequence set forth in SEQ ID NO: 148 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 33, 34, and 35, respectively; and VL amino acid sequence set forth in SEQ ID NO: 143 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 37, 38, and 39, respectively);

H) huAb13v8 (VH amino acid sequence set forth in SEQ ID NO: 148 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 33, 34, and 35, respectively; and VL amino acid sequence set forth in SEQ ID NO: 144 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 37, 38, and 39, respectively);

I) huAb13v9 (VH amino acid sequence set forth in SEQ ID NO: 148 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 33, 34, and 35, respectively; and VL amino acid sequence set forth in SEQ ID NO: 145 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 37, 38, and 39, respectively).

Thus, in one aspect the present invention provides antibodies comprising variable and/or CDR sequences from a humanized antibody derived from chAb13. In one embodiment, the invention features anti-B7-H3 antibodies which are derived from chAb13 have improved characteristics, e.g., improved binding affinity to isolated B7-H3 protein and improved binding to B7-H3 expressing cells, as described in the Examples below. Collectively these novel antibodies are referred to herein as “Ab13 variant antibodies.” Generally, the Ab13 variant antibodies retain the same epitope specificity as Ab13. In various embodiments, anti-B7-H3 antibodies, or antigen binding fragments thereof, of the invention are capable of modulating a biological function of B7-H3.

In one aspect, the present invention provides a humanized antibody, or antigen binding portion thereof, having a heavy chain variable region including an amino acid sequence set forth in SEQ ID NOs: 146, 147, or 148; and/or a light chain variable region including an amino acid sequence set forth in SEQ ID NOs: 143, 144, or 145.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen binding portion thereof, of the invention comprises a heavy chain variable region comprising a CDR1 domain comprising an amino acid sequence as set forth in SEQ ID NO: 33; a CDR2 domain comprising an amino acid sequence as set forth in SEQ ID NO: 34; and a CDR3 domain comprising an amino acid sequence as set forth in SEQ ID NO: 35; and a light chain variable region comprising a CDR1 domain comprising an amino acid sequence as set forth in SEQ ID NO: 37; a CDR2 domain comprising an amino acid sequence as set forth in SEQ ID NO: 38; and a CDR3 domain comprising an amino acid sequence as set forth in SEQ ID NO: 39.

In one aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable region including an amino acid sequence as set forth in SEQ ID NO: 147, and a light chain variable region including an amino acid sequence set forth in SEQ ID NO: 144. In one embodiment, the invention provides an anti-B7H3 antibody comprising the CDR sequences set forth in the variable regions of huAb13v1 (SEQ ID NOs. 144 and 147).

In one aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen binding portion thereof, having a heavy chain comprising the amino acid sequence of SEQ ID NO: 168 and a light chain comprising the amino acid sequence of SEQ ID NO: 169.

›DETAILED DESCRIPTION OF THE INVENTION · 18 of 40

Humanized Anti-B7-H3 Antibodies Derived from chAb18

The ten different humanized antibodies created based on chAb18 include the following:

A) huAb18v1 (VH amino acid sequence set forth in SEQ ID NO: 116 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 25, 26, and 27, respectively; and VL amino acid sequence set forth in SEQ ID NO: 120 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 29, 30, and 31, respectively);

B) huAb18v2 (VH amino acid sequence set forth in SEQ ID NO: 118 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 25, 119, and 27, respectively; and VL amino acid sequence set forth in SEQ ID NO: 120 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 29, 30, and 31, respectively);

C) huAb18v3 (VH amino acid sequence set forth in SEQ ID NO: 117 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 25, 26, and 27, respectively; and VL amino acid sequence set forth in SEQ ID NO: 121 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 29, 30, and 31, respectively);

D) huAb18v4 (VH amino acid sequence set forth in SEQ ID NO: 118 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 25, 119, and 27, respectively; and VL amino acid sequence set forth in SEQ ID NO: 121 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 29, 30, and 31, respectively);

E) huAb18v5 (VH amino acid sequence set forth in SEQ ID NO: 116 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 25, 26, and 27, respectively; and VL amino acid sequence set forth in SEQ ID NO: 123 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 29, 30, and 31, respectively);

F) huAb18v6 (VH amino acid sequence set forth in SEQ ID NO: 118 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 25, 119, and 27, respectively; and VL amino acid sequence set forth in SEQ ID NO: 123 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 29, 30, and 31, respectively);

G) huAb18v7 (VH amino acid sequence set forth in SEQ ID NO: 118 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 25, 119, and 27, respectively; and VL amino acid sequence set forth in SEQ ID NO: 124 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 29, 30, and 31, respectively);

H) huAb18v8 (VH amino acid sequence set forth in SEQ ID NO: 117 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 25, 26, and 27, respectively; and VL amino acid sequence set forth in SEQ ID NO: 122 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 29, 30, and 31, respectively);

I) huAb18v9 (VH amino acid sequence set forth in SEQ ID NO: 117 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 25, 26, and 27, respectively; and VL amino acid sequence set forth in SEQ ID NO: 124 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 29, 30, and 31, respectively); and

J) huAb18v10 (VH amino acid sequence set forth in SEQ ID NO: 118 and VH CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 25, 119, and 27, respectively; and VL amino acid sequence set forth in SEQ ID NO: 122 and VL CDR1, CDR2, and CDR3 amino acid sequences set forth in SEQ ID NOs: 29, 30, and 31, respectively).

Thus, in one aspect the present invention provides antibodies comprising variable and/or CDR sequences from a humanized antibody derived from chAb18. In one embodiment, the invention features anti-B7-H3 antibodies which are derived from Ab18 have improved characteristics, e.g., improved binding affinity to isolated B7-H3 protein and improved binding to B7-H3 expressing cells, as described in the Examples below. Collectively these novel antibodies are referred to herein as “Ab18 variant antibodies.” Generally, the Ab18 variant antibodies retain the same epitope specificity as Ab18. In various embodiments, anti-B7-H3 antibodies, or antigen binding fragments thereof, of the invention are capable of modulating a biological function of B7-H3.

›DETAILED DESCRIPTION OF THE INVENTION · 19 of 40

In one aspect, the present invention provides a humanized antibody, or antigen binding portion thereof, having a heavy chain variable region including an amino acid sequence set forth in SEQ ID NOs: 116, 117, or 118; and/or a light chain variable region including an amino acid sequence set forth in SEQ ID NOs: 120, 121, 122, 123 or 124.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen binding portion thereof, of the invention comprises a heavy chain variable region comprising a CDR1 domain comprising an amino acid sequence as set forth in SEQ ID NO: 25; a CDR2 domain comprising an amino acid sequence as set forth in SEQ ID NO: 26 or 119; and a CDR3 domain comprising an amino acid sequence as set forth in SEQ ID NO: 27; and a light chain variable region comprising a CDR1 domain comprising an amino acid sequence as set forth in SEQ ID NO: 29; a CDR2 domain comprising an amino acid sequence as set forth in SEQ ID NO: 30; and a CDR3 domain comprising an amino acid sequence as set forth in SEQ ID NO: 31.

In another aspect, the present invention is directed to a humanized anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 25; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 26; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 27; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 29; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 30; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 31.

In another aspect, the present invention is directed to a humanized anti-B7-H3 antibody, or antigen-binding portion thereof, having a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 25; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 119; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 27; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 29; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 30; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 31.

In one aspect, the present invention provides a humanized antibody, or antigen binding portion thereof, having a heavy chain variable region including an amino acid sequence set forth in SEQ ID NOs: 116, 117, 118, 146, 147, 148, 125, 126, 127, 131, 139, or 141; and/or a light chain variable region including an amino acid sequence set forth in SEQ ID NOs: 120, 121, 122, 123, 124, 143, 144, 145, 128, 129, 130, 133, 135, or 137.

In another aspect, the present invention is directed to an anti-B7-H3 antibody, or antigen binding portion thereof, of the invention comprises a heavy chain variable region comprising a CDR1 domain comprising an amino acid sequence as set forth in SEQ ID NO: 10, 25, or 33; a CDR2 domain comprising an amino acid sequence as set forth in SEQ ID NO: 11, 26, 34, 119, 132, 140, or 142; and a CDR3 domain comprising an amino acid sequence as set forth in SEQ ID NO: 12, 27, or 35; and a light chain variable region comprising a CDR1 domain comprising an amino acid sequence as set forth in SEQ ID NO: 14, 29, 37, 134, 136, or 138; a CDR2 domain comprising an amino acid sequence as set forth in SEQ ID NO: 7, 30, or 38; and a CDR3 domain comprising an amino acid sequence as set forth in SEQ ID NO: 15, 31 or 39.

›DETAILED DESCRIPTION OF THE INVENTION · 20 of 40

In another aspect, the invention provides an anti-B7-H3 antibody, or antigen binding fragment thereof, that specifically competes with an anti-B7-H3 antibody, or fragment thereof, as described herein, wherein said competition can be detected in a competitive binding assay using said antibody, the human B7-H3 polypeptide, and the anti-B7-H3 antibody or fragment thereof.

In particular embodiments, the competing antibody, or antigen binding portion thereof, is an antibody, or antigen binding portion thereof, that competes with huAb3v2.5, huAb3v2.6, or huAb13v1.

In one embodiment, the anti-B7-H3 antibodies, or antigen binding portions thereof, of the invention bind to the extracellular domain of human B7-H3 (SEQ ID NO: 152) with a dissociation constant (K D ) of about 1×10 −6 M or less, as determined by surface plasmon resonance. Alternatively, the antibodies, or antigen binding portions thereof, bind to human B7-H3 with a K D of between about 1×10 −6 M and about 1×10 −11 M, as determined by surface plasmon resonance. In a further alternative, antibodies, or antigen binding portions thereof, bind to human B7-H3 with a K D of between about 1×10 −6 M and about 1×10 −7 M, as determined by surface plasmon resonance. Alternatively, antibodies, or antigen binding portions thereof, of the invention binds to human B7-H3 with a K D of between about 1×10 −6 M and about 5×10 −11 M, about 1×10 −6 M and about 5×10 −1 0 M; a K D of between about 1×10 −6 M and about 1×10 −9 M; a K D of between about 1×10 −6 M and about 5×10 −9 M; a K D of between about 1×10 −6 M and about 1×10 −8 M; a K D of between about 1×10 −6 M and about 5×10 −8 M; a K D of between about 1×10 −7 M and about 3.4×10 −11 M; a K D of between about 5.9×10 −7 M; and about 2.2×10 −7 M, as determined by surface plasmon resonance.

In one embodiment, the antibodies, or antigen binding portions thereof, of the invention bind to human B7-H3 (SEQ ID NO: 149) with a K D of about 1×10 −6 M or less, as determined by surface plasmon resonance. Alternatively, the antibodies, or antigen binding portions thereof, of the invention bind to human B7-H3 (SEQ ID NO: 149) with a K D of between about 8.2×10 −9 M and about 6.3×10 −10 M; a K D of between about 8.2×10 −9 M and about 2.0×10 −9 M; a K D of between about 2.3×10 −9 M and about 1.5×10 −10 M, as determined by surface plasmon resonance.

The foregoing establish a novel family of B7-H3 binding proteins, isolated in accordance with this invention, and including antigen binding polypeptides that comprise the CDR sequences listed in the Sequence Table provided herein.

Anti-B7-H3 antibodies provided herein may comprise a heavy chain variable region comprising CDR1, CDR2 and CDR3 sequences and a light chain variable region comprising CDR1, CDR2 and CDR3 sequences, wherein one or more of these CDR sequences comprise specified amino acid sequences based on the antibodies described herein (e.g., huAb13v1 or huAb3v2.5), or conservative modifications thereof, and wherein the antibodies retain the desired functional properties of the anti-B7-H3antibodies described herein. Accordingly, the anti-B7-H3 antibody, or antigen binding portion thereof, may comprise a heavy chain variable region comprising CDR1, CDR2, and CDR3 sequences and a light chain variable region comprising CDR1, CDR2, and CDR3 sequences, wherein: (a) the heavy chain variable region CDR3 sequence comprises SEQ ID NO: 12 or 35, and conservative modifications thereof, e.g., 1, 2, 3, 4, 5, 1-2, 1-3, 1-4 or 1-5 conservative amino acid substitutions; (b) the light chain variable region CDR3 sequence comprises SEQ ID NO: 15 or 39, and conservative modifications thereof, e.g., 1, 2, 3, 4, 5, 1-2, 1-3, 1-4 or 1-5 conservative amino acid substitutions; (c) the antibody specifically binds to B7-H3, and (d) the antibody exhibits 1, 2, 3, 4, 5, 6, or all of the following functional properties described herein, e.g., binding to soluble human B7-H3. In a one embodiment, the heavy chain variable region CDR2 sequence comprises SEQ ID NO: 140 or 34, and conservative modifications thereof, e.g., 1, 2, 3, 4, 5, 1-2, 1-3, 1-4 or 1-5 conservative amino acid substitutions; and the light chain variable region CDR2 sequence comprises SEQ ID NO: 7 or 38, and conservative modifications thereof, e.g., 1, 2, 3, 4, 5, 1-2, 1-3, 1-4 or 1-5 conservative amino acid substitutions. In another preferred embodiment, the heavy chain variable region CDR1 sequence comprises SEQ ID NO: 10 or 33, and conservative modifications thereof, e.g., 1, 2, 3, 4, 5, 1-2, 1-3, 1-4 or 1-5 conservative amino acid substitutions; and the light chain variable region CDR1 sequence comprises SEQ ID NO: 136, 138, or 37, and conservative modifications thereof, e.g., 1, 2, 3, 4, 5, 1-2, 1-3, 1-4 or 1-5 conservative amino acid substitutions.

Conservative amino acid substitutions may also be made in portions of the antibodies other than, or in addition to, the CDRs. For example, conservative amino acid modifications may be made in a framework region or in the Fc region. A variable region or a heavy or light chain may comprise 1, 2, 3, 4, 5, 1-2, 1-3, 1-4, 1-5, 1-10, 1-15, 1-20, 1-25, or 1-50 conservative amino acid substitutions relative to the anti-B7-H3 antibody sequences provided herein. In certain embodiments, the anti-B7-H3antibody comprises a combination of conservative and non-conservative amino acid modification.

To generate and to select CDRs having preferred B7-H3 binding and/or neutralizing activity with respect to hB7-H3, standard methods known in the art for generating antibodies, or antigen binding portions thereof, and assessing the B7-H3 binding and/or neutralizing characteristics of those antibodies, or antigen binding portions thereof, may be used, including but not limited to those specifically described herein.

In certain embodiments, the antibody comprises a heavy chain constant region, such as an IgG1, IgG2, IgG3, IgG4, IgA, IgE, IgM, or IgD constant region. In certain embodiments, the anti-B7-H3 antibody, or antigen binding portion thereof, comprises a heavy chain immunoglobulin constant domain selected from the group consisting of a human IgG constant domain, a human IgM constant domain, a human IgE constant domain, and a human IgA constant domain. In further embodiments, the antibody, or antigen binding portion thereof, has an IgG1 heavy chain constant region, an IgG2 heavy chain constant region, an IgG3 constant region, or an IgG4 heavy chain constant region. Preferably, the heavy chain constant region is an IgG1 heavy chain constant region or an IgG4 heavy chain constant region. Furthermore, the antibody can comprise a light chain constant region, either a kappa light chain constant region or a lambda light chain constant region. Preferably, the antibody comprises a kappa light chain constant region. Alternatively, the antibody portion can be, for example, a Fab fragment or a single chain Fv fragment.

›DETAILED DESCRIPTION OF THE INVENTION · 21 of 40

In certain embodiments, the anti-B7-H3 antibody binding portion is a Fab, a Fab′, a F(ab′)2, a Fv, a disulfide linked Fv, an scFv, a single domain antibody, or a diabody.

In certain embodiments, the anti-B7-H3 antibody, or antigen binding portion thereof, is a multispecific antibody, e.g. a bispecific antibody.

Replacements of amino acid residues in the Fc portion to alter antibody effector function have been described (Winter, et al. U.S. Pat. Nos. 5,648,260 and 5,624,821, incorporated by reference herein). The Fc portion of an antibody mediates several important effector functions e.g. cytokine induction, ADCC, phagocytosis, complement dependent cytotoxicity (CDC) and half-life/clearance rate of antibody and antigen-antibody complexes. In some cases these effector functions are desirable for therapeutic antibody but in other cases might be unnecessary or even deleterious, depending on the therapeutic objectives. Certain human IgG isotypes, particularly IgG1 and IgG3, mediate ADCC and CDC via binding to FcγRs and complement C1q, respectively. Neonatal Fc receptors (FcRn) are the critical components determining the circulating half-life of antibodies. In still another embodiment at least one amino acid residue is replaced in the constant region of the antibody, for example the Fc region of the antibody, such that effector functions of the antibody are altered.

One embodiment of the invention includes a recombinant chimeric antigen receptor (CAR) comprising the binding regions of the antibodies described herein, e.g., the heavy and/or light chain CDRs of huAb13v1. A recombinant CAR, as described herein, may be used to redirect T cell specificity to an antigen in a human leukocyte antigen (HLA)-independent fashion. Thus, CARs of the invention may be used in immunotherapy to help engineer a human subject's own immune cells to recognize and attack the subject's tumor (see, e.g., U.S. Pat. Nos. 6,410,319; 8,389,282; 8,822,647; 8,906,682; 8,911,993; 8,916,381; 8,975,071; and U.S. Patent Appln. Publ. No. US20140322275, each of which is incorporated by reference herein with respect to CAR technology). This type of immunotherapy is called adoptive cell transfer (ACT), and may be used to treat cancer in a subject in need thereof.

An anti-B7-H3 CAR of the invention preferably contains a extracellular antigen-binding domain specific for B7-H3, a transmembrane domain which is used to anchor the CAR into a T cell, and one or more intracellular signaling domains. In one embodiment of the invention, the CAR includes a transmembrane domain that comprises a transmembrane domain of a protein selected from the group consisting of the alpha, beta or zeta chain of the T-cell receptor, CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137 and CD154. In one embodiment of the invention, the CAR comprises a costimulatory domain, e.g., a costimulatory domain comprising a functional signaling domain of a protein selected from the group consisting of OX40, CD2, CD27, CD28, CD5, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278), and 4-1BB (CD137). In certain embodiments of the invention, the CAR comprises an scFv comprising the CDR or variable regions described herein e.g., CDRs or variable regions from the huAb13v1 antibody, a transmembrane domain, a co-stimulatory domain (e.g., a functional signaling domain from CD28 or 4-1BB), and a signaling domain comprising a functional signaling domain from CD3 (e.g., CD3-zeta).

In certain embodiments, the invention incudes a T cell comprising a CAR (also referred to as a CAR T cell) comprising antigen binding regions, e.g. CDRs, of the antibodies described herein or an scFv described herein.

In certain embodiments of the invention, the CAR comprises a heavy chain variable region comprising a CDR1 domain comprising an amino acid sequence as set forth in SEQ ID NO: 10, 25, or 33; a CDR2 domain comprising an amino acid sequence as set forth in SEQ ID NO: 11, 26, 34, 119, 132, 140, or 142; and a CDR3 domain comprising an amino acid sequence as set forth in SEQ ID NO: 12, 27, or 35; and a light chain variable region comprising a CDR1 domain comprising an amino acid sequence as set forth in SEQ ID NO: 14, 29, 37, 134, 136, or 138; a CDR2 domain comprising an amino acid sequence as set forth in SEQ ID NO: 7, 30, or 38; and a CDR3 domain comprising an amino acid sequence as set forth in SEQ ID NO: 15, 31 or 39.

In certain embodiments of the invention, the CAR comprises a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 10; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 11; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 12; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 14; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 7; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 15.

In certain embodiments of the invention, the CAR comprises a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 10; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 132; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 12; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 134; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 7; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 15.

In certain embodiments of the invention, the CAR comprises a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 10; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 132; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 12; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 136; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 7; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 15.

›DETAILED DESCRIPTION OF THE INVENTION · 22 of 40

In certain embodiments of the invention, the CAR comprises a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 10; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 132; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 12; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 138; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 7; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 15.

In certain embodiments of the invention, the CAR comprises a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 10; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 140; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 12; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 134; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 7; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 15.

In certain embodiments of the invention, the CAR comprises a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 10; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 140; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 12; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 136; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 7; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 15.

In certain embodiments of the invention, the CAR comprises a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 10; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 140; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 12; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 138; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 7; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 15.

In certain embodiments of the invention, the CAR comprises a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 10; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 142; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 12; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 134; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 7; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 15.

In certain embodiments of the invention, the CAR comprises a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 10; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 142; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 12; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 136; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 7; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 15.

In certain embodiments of the invention, the CAR comprises a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 10; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 142; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 12; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 138; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 7; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 15.

In certain embodiments of the invention, the CAR comprises a heavy chain variable region comprising a CDR1 domain comprising an amino acid sequence as set forth in SEQ ID NO: 33; a CDR2 domain comprising an amino acid sequence as set forth in SEQ ID NO: 34; and a CDR3 domain comprising an amino acid sequence as set forth in SEQ ID NO: 35; and a light chain variable region comprising a CDR1 domain comprising an amino acid sequence as set forth in SEQ ID NO: 37; a CDR2 domain comprising an amino acid sequence as set forth in SEQ ID NO: 38; and a CDR3 domain comprising an amino acid sequence as set forth in SEQ ID NO: 39.

In certain embodiments of the invention, the CAR comprises a heavy chain variable region comprising a CDR1 domain comprising an amino acid sequence as set forth in SEQ ID NO: 25; a CDR2 domain comprising an amino acid sequence as set forth in SEQ ID NO: 26 or 119; and a CDR3 domain comprising an amino acid sequence as set forth in SEQ ID NO: 27; and a light chain variable region comprising a CDR1 domain comprising an amino acid sequence as set forth in SEQ ID NO: 29; a CDR2 domain comprising an amino acid sequence as set forth in SEQ ID NO: 30; and a CDR3 domain comprising an amino acid sequence as set forth in SEQ ID NO: 31.

In certain embodiments of the invention, the CAR comprises a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 25; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 26; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 27; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 29; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 30; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 31.

In certain embodiments of the invention, the CAR comprises a heavy chain variable domain region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 25; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 119; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 27; and a light chain variable region including (a) a CDR1 having an amino acid sequence as set forth in SEQ ID NO: 29; (b) a CDR2 having an amino acid sequence as set forth in SEQ ID NO: 30; and (c) a CDR3 having an amino acid sequence as set forth in SEQ ID NO: 31.

›DETAILED DESCRIPTION OF THE INVENTION · 23 of 40

One embodiment of the invention includes a labeled anti-B7-H3 antibody, or antibody portion thereof, where the antibody is derivatized or linked to one or more functional molecule(s) (e.g., another peptide or protein). For example, a labeled antibody can be derived by functionally linking an antibody or antibody portion of the invention (by chemical coupling, genetic fusion, noncovalent association or otherwise) to one or more other molecular entities, such as another antibody (e.g., a bispecific antibody or a diabody), a detectable agent, a pharmaceutical agent, a protein or peptide that can mediate the association of the antibody or antibody portion with another molecule (such as a streptavidin core region or a polyhistidine tag), and/or a cytotoxic or therapeutic agent selected from the group consisting of a mitotic inhibitor, an antitumor antibiotic, an immunomodulating agent, a vector for gene therapy, an alkylating agent, an antiangiogenic agent, an antimetabolite, a boron-containing agent, a chemoprotective agent, a hormone, an antihormone agent, a corticosteroid, a photoactive therapeutic agent, an oligonucleotide, a radionuclide agent, a topoisomerase inhibitor, a kinase inhibitor, a radiosensitizer, and a combination thereof.

Useful detectable agents with which an antibody or antibody portion thereof, may be derivatized include fluorescent compounds. Exemplary fluorescent detectable agents include fluorescein, fluorescein isothiocyanate, rhodamine, 5-dimethylamine-1-napthalenesulfonyl chloride, phycoerythrin and the like. An antibody may also be derivatized with detectable enzymes, such as alkaline phosphatase, horseradish peroxidase, glucose oxidase and the like. When an antibody is derivatized with a detectable enzyme, it is detected by adding additional reagents that the enzyme uses to produce a detectable reaction product. For example, when the detectable agent horseradish peroxidase is present the addition of hydrogen peroxide and diaminobenzidine leads to a colored reaction product, which is detectable. An antibody may also be derivatized with biotin, and detected through indirect measurement of avidin or streptavidin binding.

In one embodiment, the antibody of the invention is conjugated to an imaging agent. Examples of imaging agents that may be used in the compositions and methods described herein include, but are not limited to, a radiolabel (e.g., indium), an enzyme, a fluorescent label, a luminescent label, a bioluminescent label, a magnetic label, and biotin.

In one embodiment, the antibodies or ADCs are linked to a radiolabel, such as, but not limited to, indium ( 111 In). 111 Indium may be used to label the antibodies and ADCs described herein for use in identifying B7-H3 positive tumors. In a certain embodiment, anti-B7-H3 antibodies (or ADCs) described herein are labeled with 111 I via a bifunctional chelator which is a bifunctional cyclohexyl diethylenetriaminepentaacetic acid (DTPA) chelate (see U.S. Pat. Nos. 5,124,471; 5,434,287; and 5,286,850, each of which is incorporated herein by reference).

Another embodiment of the invention provides a glycosylated binding protein wherein the anti-B7-H3 antibody or antigen binding portion thereof comprises one or more carbohydrate residues. Nascent in vivo protein production may undergo further processing, known as post-translational modification. In particular, sugar (glycosyl) residues may be added enzymatically, a process known as glycosylation. The resulting proteins bearing covalently linked oligosaccharide side chains are known as glycosylated proteins or glycoproteins. Antibodies are glycoproteins with one or more carbohydrate residues in the Fc domain, as well as the variable domain. Carbohydrate residues in the Fc domain have important effect on the effector function of the Fc domain, with minimal effect on antigen binding or half-life of the antibody (R. Jefferis, Biotechnol. Prog. 21 (2005), pp. 11-16). In contrast, glycosylation of the variable domain may have an effect on the antigen binding activity of the antibody. Glycosylation in the variable domain may have a negative effect on antibody binding affinity, likely due to steric hindrance (Co, M. S., et al., Mol. Immunol . (1993) 30:1361-1367), or result in increased affinity for the antigen (Wallick, S. C., et al., Exp. Med . (1988) 168:1099-1109; Wright, A., et al., EMBO J . (1991) 10:2717-2723).

One aspect of the invention is directed to generating glycosylation site mutants in which the O- or N-linked glycosylation site of the binding protein has been mutated. One skilled in the art can generate such mutants using standard well-known technologies. Glycosylation site mutants that retain the biological activity, but have increased or decreased binding activity, are another object of the invention.

In still another embodiment, the glycosylation of the anti-B7-H3 antibody or antigen binding portion of the invention is modified. For example, an aglycoslated antibody can be made (i.e., the antibody lacks glycosylation). Glycosylation can be altered to, for example, increase the affinity of the antibody for antigen. Such carbohydrate modifications can be accomplished by, for example, altering one or more sites of glycosylation within the antibody sequence. For example, one or more amino acid substitutions can be made that result in elimination of one or more variable region glycosylation sites to thereby eliminate glycosylation at that site. Such aglycosylation may increase the affinity of the antibody for antigen. Such an approach is described in further detail in PCT Publication WO2003016466A2, and U.S. Pat. Nos. 5,714,350 and 6,350,861, each of which is incorporated herein by reference in its entirety.

Additionally or alternatively, a modified anti-B7-H3 antibody of the invention can be made that has an altered type of glycosylation, such as a hypofucosylated antibody having reduced amounts of fucosyl residues or an antibody having increased bisecting GlcNAc structures. Such altered glycosylation patterns have been demonstrated to increase the ADCC ability of antibodies. Such carbohydrate modifications can be accomplished by, for example, expressing the antibody in a host cell with altered glycosylation machinery. Cells with altered glycosylation machinery have been described in the art and can be used as host cells in which to express recombinant antibodies of the invention to thereby produce an antibody with altered glycosylation. See, for example, Shields, R. L. et al. (2002) J. Biol. Chem. 277:26733-26740; Umana et al. (1999) Nat. Biotech. 17:176-1, as well as, European Patent No: EP 1,176,195; PCT Publications WO 03/035835; WO 99/54342 80, each of which is incorporated herein by reference in its entirety.

›DETAILED DESCRIPTION OF THE INVENTION · 24 of 40

Protein glycosylation depends on the amino acid sequence of the protein of interest, as well as the host cell in which the protein is expressed. Different organisms may produce different glycosylation enzymes (e.g., glycosyltransferases and glycosidases), and have different substrates (nucleotide sugars) available. Due to such factors, protein glycosylation pattern, and composition of glycosyl residues, may differ depending on the host system in which the particular protein is expressed. Glycosyl residues useful in the invention may include, but are not limited to, glucose, galactose, mannose, fucose, n-acetylglucosamine and sialic acid. Preferably the glycosylated binding protein comprises glycosyl residues such that the glycosylation pattern is human.

Differing protein glycosylation may result in differing protein characteristics. For instance, the efficacy of a therapeutic protein produced in a microorganism host, such as yeast, and glycosylated utilizing the yeast endogenous pathway may be reduced compared to that of the same protein expressed in a mammalian cell, such as a CHO cell line. Such glycoproteins may also be immunogenic in humans and show reduced half-life in vivo after administration. Specific receptors in humans and other animals may recognize specific glycosyl residues and promote the rapid clearance of the protein from the bloodstream. Other adverse effects may include changes in protein folding, solubility, susceptibility to proteases, trafficking, transport, compartmentalization, secretion, recognition by other proteins or factors, antigenicity, or allergenicity. Accordingly, a practitioner may prefer a therapeutic protein with a specific composition and pattern of glycosylation, for example glycosylation composition and pattern identical, or at least similar, to that produced in human cells or in the species-specific cells of the intended subject animal.

Expressing glycosylated proteins different from that of a host cell may be achieved by genetically modifying the host cell to express heterologous glycosylation enzymes. Using recombinant techniques, a practitioner may generate antibodies or antigen binding portions thereof exhibiting human protein glycosylation. For example, yeast strains have been genetically modified to express non-naturally occurring glycosylation enzymes such that glycosylated proteins (glycoproteins) produced in these yeast strains exhibit protein glycosylation identical to that of animal cells, especially human cells (U.S. patent Publication Nos. 20040018590 and 20020137134 and PCT publication WO2005100584 A2).

Antibodies may be produced by any of a number of techniques. For example, expression from host cells, wherein expression vector(s) encoding the heavy and light chains is (are) transfected into a host cell by standard techniques. The various forms of the term “transfection” are intended to encompass a wide variety of techniques commonly used for the introduction of exogenous DNA into a prokaryotic or eukaryotic host cell, e.g., electroporation, calcium-phosphate precipitation, DEAE-dextran transfection and the like. Although it is possible to express antibodies in either prokaryotic or eukaryotic host cells, expression of antibodies in eukaryotic cells is preferable, and most preferable in mammalian host cells, because such eukaryotic cells (and in particular mammalian cells) are more likely than prokaryotic cells to assemble and secrete a properly folded and immunologically active antibody.

Preferred mammalian host cells for expressing the recombinant antibodies of the invention include Chinese Hamster Ovary (CHO cells) (including dhfr-CHO cells, described in Urlaub and Chasin, (1980) Proc. Natl. Acad. Sci. USA 77:4216-4220, used with a DHFR selectable marker, e.g., as described in R. J. Kaufman and P. A. Sharp (1982) Mol. Biol. 159:601-621), NS0 myeloma cells, COS cells and SP2 cells. When recombinant expression vectors encoding antibody genes are introduced into mammalian host cells, the antibodies are produced by culturing the host cells for a period of time sufficient to allow for expression of the antibody in the host cells or, more preferably, secretion of the antibody into the culture medium in which the host cells are grown. Antibodies can be recovered from the culture medium using standard protein purification methods.

Host cells can also be used to produce functional antibody fragments, such as Fab fragments or scFv molecules. It will be understood that variations on the above procedure are within the scope of the invention. For example, it may be desirable to transfect a host cell with DNA encoding functional fragments of either the light chain and/or the heavy chain of an antibody of this invention. Recombinant DNA technology may also be used to remove some, or all, of the DNA encoding either or both of the light and heavy chains that is not necessary for binding to the antigens of interest. The molecules expressed from such truncated DNA molecules are also encompassed by the antibodies of the invention. In addition, bifunctional antibodies may be produced in which one heavy and one light chain are an antibody of the invention and the other heavy and light chain are specific for an antigen other than the antigens of interest by crosslinking an antibody of the invention to a second antibody by standard chemical crosslinking methods.

In a preferred system for recombinant expression of an antibody, or antigen binding portion thereof, a recombinant expression vector encoding both the antibody heavy chain and the antibody light chain is introduced into dhfr-CHO cells by calcium phosphate-mediated transfection. Within the recombinant expression vector, the antibody heavy and light chain genes are each operatively linked to CMV enhancer/AdMLP promoter regulatory elements to drive high levels of transcription of the genes. The recombinant expression vector also carries a DHFR gene, which allows for selection of CHO cells that have been transfected with the vector using methotrexate selection/amplification. The selected transformant host cells are cultured to allow for expression of the antibody heavy and light chains and intact antibody is recovered from the culture medium. Standard molecular biology techniques are used to prepare the recombinant expression vector, transfect the host cells, select for transformants, culture the host cells and recover the antibody from the culture medium. Still further the invention provides a method of synthesizing a recombinant antibody of the invention by culturing a host cell in a suitable culture medium until a recombinant antibody is synthesized. Recombinant antibodies of the invention may be produced using nucleic acid molecules corresponding to the amino acid sequences disclosed herein The method can further comprise isolating the recombinant antibody from the culture medium.

›DETAILED DESCRIPTION OF THE INVENTION · 25 of 40

The N- and C-termini of antibody polypeptide chains of the present invention may differ from the expected sequence due to commonly observed post-translational modifications. For example, C-terminal lysine residues are often missing from antibody heavy chains. Dick et al. (2008) Biotechnol. Bioeng. 100:1132. N-terminal glutamine residues, and to a lesser extent glutamate residues, are frequently converted to pyroglutamate residues on both light and heavy chains of therapeutic antibodies. Dick et al. (2007) Biotechnol. Bioeng. 97:544; Liu et al. (2011) JBC 28611211; Liu et al. (2011) J. Biol. Chem. 286:11211.

III. Anti-B7-H3 Antibody Drug Conjugates (ADCs)

Anti-B7-H3 antibodies described herein may be conjugated to a drug moiety to form an anti-B7-H3 Antibody Drug Conjugate (ADC). Antibody-drug conjugates (ADCs) may increase the therapeutic efficacy of antibodies in treating disease, e.g., cancer, due to the ability of the ADC to selectively deliver one or more drug moiety(s) to target tissues, such as a tumor-associated antigen, e.g., B7-H3 expressing tumors. Thus, in certain embodiments, the invention provides anti-B7-H3 ADCs for therapeutic use, e.g., treatment of cancer.

Anti-B7-H3 ADCs of the invention comprise an anti-B7-H3 antibody, i.e., an antibody that specifically binds to B7-H3, linked to one or more drug moieties. The specificity of the ADC is defined by the specificity of the antibody, i.e., anti-B7-H3. In one embodiment, an anti-B7-H3 antibody is linked to one or more cytotoxic drug(s) which is delivered internally to a transformed cancer cell expressing B7-H3.

Examples of drugs that may be used in the anti-B7-H3 ADC of the invention are provided below, as are linkers that may be used to conjugate the antibody and the one or more drug(s). The terms “drug,” “agent,” and “drug moiety” are used interchangeably herein. The terms “linked” and “conjugated” are also used interchangeably herein and indicate that the antibody and moiety are covalently linked.

In some embodiments, the ADC has the following formula (formula I):

(D-L-LK m Ab (I)

wherein Ab is the antibody, e.g., anti-B7-H3 antibody huAb13v1, huAb3v2.5, or huAb3v2.6, and (L) is a linker, (D) is a drug, and LK represents a covalent linkage linking linker L to antibody Ab; and m is an integer ranging from 1 to 20. D is a drug moiety having, for example, cytostatic, cytotoxic, or otherwise therapeutic activity against a target cell, e.g., a cell expressing B7-H3. In some embodiments, m ranges from 1 to 8, 1 to 7, 1 to 6, 2 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 1.5 to 8, 1.5 to 7, 1.5 to 6, 1.5 to 5, 1.5 to 4, 2 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2,or 2 to 4. The DAR of an ADC is equivalent to the “m” referred to in Formula I. In one embodiment, the ADC has a formula of Ab-(LK-L-D) m , wherein Ab is an anti-B7-H3 antibody, e.g. huAb13v1, huAb3v2.5, or huAb3v2.6, L is a linker, D is a drug, e.g., a Bcl-xL inhibitor or an auristatin such as MMAF or MMAE, and m is 2 to 4 (equivalent to a DAR of 2-4). Additional details regarding drugs (D of Formula I) and linkers (L of Formula I) that may be used in the ADCs of the invention, as well as alternative ADC structures, are described below.

III. A. Anti-B7-H3 ADCs: Bcl-xL Inhibitors, Linkers, Synthons, and Methods of Making Same

Dysregulated apoptotic pathways have also been implicated in the pathology of cancer. The implication that down-regulated apoptosis (and more particularly the Bcl-2 family of proteins) is involved in the onset of cancerous malignancy has revealed a novel way of targeting this still elusive disease. Research has shown, for example, the anti-apoptotic proteins, Bcl 2 and Bcl-xL, are over-expressed in many cancer cell types. See, Zhang, 2002, Nature Reviews/Drug Discovery 1:101; Kirkin et al., 2004, Biochimica Biophysica Acta 1644:229-249; and Amundson et al., 2000, Cancer Research 60:6101-6110. The effect of this deregulation is the survival of altered cells which would otherwise have undergone apoptosis in normal conditions. The repetition of these defects associated with unregulated proliferation is thought to be the starting point of cancerous evolution.

Aspects of the disclosure concern anti-hB7-H3 ADCs comprising an anti-hB7-H3 antibody conjugated to a drug via a linker, wherein the drug is a Bcl-xL inhibitor. In specific embodiments, the ADCs are compounds according to structural formula (I) below, or a pharmaceutically acceptable salt thereof, wherein Ab represents the anti-hB7-H3 antibody, D represents a Bcl-xL inhibitor drug (i.e., a compound of formula (IIa), (IIb), (IIc), or (IId) as shown below), L represents a linker, LK represents a covalent linkage linking the linker (L) to the anti-hB7-H3 antibody (Ab) and m represents the number of D-L-LK units linked to the antibody, which is an integer ranging from 1 to 20. In certain embodiments, m is 2, 3 or 4. In some embodiments, m ranges from 1 to 8, 1 to 7, 1 to 6, 2 to 6, 1 to 5, 1 to 4, 2 to 4, 1 to 3, 1 to 2, or is 1.

(D-L-LK m Ab (I)

Specific embodiments of various Bcl-xL inhibitors per se, and various Bcl-xL inhibitors (D), linkers (L) and anti-B7-H3 antibodies (Ab) that can comprise the ADCs described herein, as well as the number of Bcl-xL inhibitors linked to the ADCs, are described in more detail below.

Examples of Bcl-xL inhibitors that may be used in the anti-B7-H3 ADC of the invention are provided below, as are linkers that may be used to conjugate the antibody and the one or more Bcl-xL inhibitor(s). The terms “linked” and “conjugated” are also used interchangeably herein and indicate that the antibody and moiety are covalently linked.

III.A.1. Bcl-xL Inhibitors

One aspect of the instant disclosure concerns Bcl-xL inhibitors that have low cell permeability. The compounds are generally heterocyclic in nature and include one or more solubilizing groups that impart the compounds with high water solubility and low cell permeability. The solubilizing groups are generally groups that are capable of hydrogen bonding, forming dipole-dipole interactions, and/or that include a polyethylene glycol polymer containing from 1 to 30 units, one or more polyols, one or more salts, or one or more groups that are charged at physiological pH.

›DETAILED DESCRIPTION OF THE INVENTION · 26 of 40

The Bcl-xL inhibitors may be used as compounds or salts per se in the various methods described herein, or may be included as a component part of an ADC.

Specific embodiments of Bcl-xL inhibitors that may be used in unconjugated form, or that may be included as part of an ADC include compounds according to structural formulae (IIa), (IIb), (IIc), or (IId). In the present invention, when the Bcl-xL inhibitors are included as part of an ADC, # shown in structural formula (IIa), (IIb), (IIc), or (IId) below represents a point of attachment to a linker, which indicates that they are represented in a monoradical form.

or a pharmaceutically acceptable salt thereof, wherein:

Ar 1 is selected from

and is optionally substituted with one or more substituents independently selected from halo, hydroxy, nitro, lower alkyl, lower heteroalkyl, C 1-4 alkoxy, amino, cyano and halomethyl;

Ar 2 is selected from

and is optionally substituted with one or more substituents independently selected from halo, hydroxy, nitro, lower alkyl, lower heteroalkyl, C 1-4 alkoxy, amino, cyano and halomethyl, wherein the R 12 —Z 2b —, R′—Z 2b- , #—N(R 4 )—R 13 —Z 2b —, or #—R′—Z 2b — substituents are attached to Ar 2 at any Ar 2 atom capable of being substituted;

Z 1 is selected from N, CH, C-halo, C—CH 3 and C—CN;

Z 2a and Z 2b are each, independently from one another, selected from a bond, NR 6 , CR 6a R 6b , O, S, S(O), S(O) 2 , —NR 6 C(O)—, —NR 6a C(O)NR 6b —, and —NR 6 C(O)O—;

R′ is a alkylene, heteroalkylene, cycloalkylene, heterocyclene, aryl or heteroaryl independently substituted at one or more carbon or heteroatoms with a solubilizing moiety containing a group selected from a polyol, a polyethylene glycol containing from 4 to 30 ethylene glycol units, a salt, and a group that is charged at physiological pH and combinations thereof, wherein #, where attached to R′, is attached to R′ at any R′ atom capable of being substituted;

R 1 is selected from hydrogen, methyl, halo, halomethyl, ethyl, and cyano;

R 2 is selected from hydrogen, methyl, halo, halomethyl and cyano;

R 3 is selected from hydrogen, methyl, ethyl, halomethyl and haloethyl;

R 4 is selected from hydrogen, lower alkyl and lower heteroalkyl or is taken together with an atom of R 13 to form a cycloalkyl or heterocyclyl ring having between 3 and 7 ring atoms;

R 6 , R 6a and R 6b are each, independent from one another, selected from hydrogen, optionally substituted lower alkyl, optionally substituted lower heteroalkyl, optionally substituted cycloalkyl and optionally substituted heterocyclyl, or are taken together with an atom from R 4 and an atom from R 13 to form a cycloalkyl or heterocyclyl ring having between 3 and 7 ring atoms;

R 11a and R 11b are each, independently of one another, selected from hydrogen, halo, methyl, ethyl, halomethyl, hydroxyl, methoxy, CN, and SCH 3 ;

R 12 is optionally R′ or is selected from hydrogen, halo, cyano, optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, and optionally substituted cycloalkyl;

R 13 is selected from optionally substituted C 1-8 alkylene, optionally substituted heteroalkylene, optionally substituted heterocyclene, and optionally substituted cycloalkylene; and

# represents the point of attachment to a linker L.

One embodiment of Bcl-xL inhibitors that may be used in unconjugated form, or that may be included as part of an ADC include compounds according to structural formulae (IIa), (IIb), (IIc), or (IId):

or a pharmaceutically acceptable salt thereof, wherein:

Ar 1 is selected from

and is optionally substituted with one or more substituents independently selected from halo, hydroxy, nitro, lower alkyl, lower heteroalkyl, C 1-4 alkoxy, amino, cyano and halomethyl;

Ar 2 is selected from

Z 1 is selected from N, CH, C-halo, C—CH 3 and C—CN;

Z 2a and Z 2b are each, independently from one another, selected from a bond, NR 6 , CR 6a R 6b , O, S, S(O), S(O) 2 , —NR 6 C(O)—, —NR 6a C(O)NR 6b —, and —NR 6 C(O)O—;

R′ is

wherein #, where attached to R′, is attached to R′ at any R′ atom capable of being substituted;

X′ is selected at each occurrence from —N(R 10 )—, —N(R 10 )C(O)—, —N(R 10 )S(O) 2 —, —S(O) 2 N(R 10 )—, and —O—;

n is selected from 0-3;

R 10 is independently selected at each occurrence from hydrogen, lower alkyl, heterocycle, aminoalkyl, G-alkyl, and —(CH 2 ) 2 —O—(CH 2 ) 2 —O—(CH 2 ) 2 —NH 2 ;

G at each occurrence is independently selected from a polyol, a polyethylene glycol with between 4 and 30 repeating units, a salt and a moiety that is charged at physiological pH;

SP a is independently selected at each occurrence from oxygen, —S(O) 2 N(H)—, —N(H)S(O) 2 —, —N(H)C(O)—, —C(O)N(H)—, —N(H)—, arylene, heterocyclene, and optionally substituted methylene; wherein methylene is optionally substituted with one or more of —NH(CH 2 ) 2 G, NH 2 , C 1-8 alkyl, and carbonyl;

m 2 is selected from 0-12;

R 1 is selected from hydrogen, methyl, halo, halomethyl, ethyl, and cyano;

R 2 is selected from hydrogen, methyl, halo, halomethyl and cyano;

R 3 is selected from hydrogen, methyl, ethyl, halomethyl and haloethyl;

R 4 is selected from hydrogen, lower alkyl and lower heteroalkyl or is taken together with an atom of R 13 to form a cycloalkyl or heterocyclyl ring having between 3 and 7 ring atoms;

›DETAILED DESCRIPTION OF THE INVENTION · 27 of 40

R 11a and R 11b are each, independently of one another, selected from hydrogen, halo, methyl, ethyl, halomethyl, hydroxyl, methoxy, CN, and SCH 3 ;

R 13 is selected from optionally substituted C 1-8 alkylene, optionally substituted heteroalkylene, optionally substituted heterocyclene, and optionally substituted cycloalkylene; and

# represents the point of attachment to a linker L.

When a Bcl-xL inhibitor of structural formulae (IIa)-(IId) is not a component of an ADC, # in formulae (IIa)-(IId) represents the point of attachment to a hydrogen atom. When the Bcl-xL inhibitor is a component of an ADC, # in formulae (IIa)-(IId) represents the point of attachment to the linker. When a Bcl-xL inhibitor is a component of an ADC, the ADC may comprise one or more Bcl-xL inhibitors, which may be the same or different, but are typically the same.

In certain embodiments, R′ is a C 2 -C 8 heteroalkylene substituted with one or more moieties containing a salt and/or a group that is charged at physiological pH. The salt may be selected, for example, from the salt of a carboxylate, a sulfonate, a phosphonate, and an ammonium ion. For example, the salt may be the sodium or potassium salt of a carboxylate, sulfonate or phosphonate or the chloride salt of an ammonium ion. The group that is charged at physiological pH may be any group that is charged at a physiological pH, including, by way of example and not limitation, a zwitterionic group. In certain embodiments a group that is a salt is a dipolar moiety such as, but not limited to, N-oxides of amines including certain heterocyclyls such as, but not limited to, pyridine and quinoline. In specific embodiments the group that is charged at physiological pH is selected independently at each occurrence, from carboxylate, sulfonate, phosphonate, and amine.

In certain embodiments, R′ is a C 2 -C 8 heteroalkylene substituted with one or more moieties containing polyethylene glycol or a polyol such as a diol or a sugar moiety.

In certain embodiments, R′ may be substituted with groups in addition to a solubilizing moiety. For example, R′ may be substituted with one or more of the same or different alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, or halo groups.

In certain embodiments, R′ is represented by the formula:

or a pharmaceutically acceptable salt thereof, wherein:

X′ is selected at each occurrence from —N(R 10 )— and —O—;

n is selected from 1-3;

R 10 is individually selected at each occurrence from hydrogen, alkyl, heterocycle, aminoalkyl, G-alkyl, heterocycle, and —(CH 2 ) 2 —O—(CH 2 ) 2 —O—(CH 2 ) 2 —NH 2 ;

G at each occurrence is independently selected from a polyol, a polyethylene glycol with between 4 and 30 repeating unit (referred to herein as PEG4-30), a salt and a moiety that is charged at physiological pH;

SP a is independently selected at each occurrence from oxygen, sulfonamide, arylene, heterocyclene, and optionally substituted methylene; wherein methylene is optionally substituted with one or more of —NH(CH 2 ) 2 G, amine and carbonyl; and

m 2 is selected from 0-6,

wherein there is at least one substitutable nitrogen in R′ that is attached to a linker or a hydrogen atom at a substitutable nitrogen atom of R′.

In certain embodiments, R′ is

X′ is selected at each occurrence from —N(R 10 )—, —N(R 10 )C(O)—, —N(R 10 )S(O) 2 —, —S(O) 2 N(R 10 )—, and —O—;

n is selected from 0-3;

R 10 is independently selected at each occurrence from hydrogen, alkyl, heterocycle, aminoalkyl, G-alkyl, heterocycle, and —(CH 2 ) 2 —O—(CH 2 ) 2 —O—(CH 2 ) 2 —NH 2 ;

G at each occurrence is independently selected from a polyol, a polyethylene glycol with between 4 and 30 repeating units, a salt and a moiety that is charged at physiological pH;

SP a is independently selected at each occurrence from oxygen-S(O) 2 N(H)—, —N(H)S(O) 2 —, —N(H)C(O)—, —C(O)N(H)—, —N(H)—, arylene, heterocyclene, and optionally substituted methylene; wherein methylene is optionally substituted with one or more of —NH(CH 2 ) 2 G, amine, alkyl, and carbonyl;

m 2 is selected from 0-12, and

#, where attached to R′, is attached to R′ at any R′ atom capable of being substituted.

In certain embodiments, G at each occurrence is a salt or a moiety that is charged at physiological pH.

In certain embodiments, G at each occurrence is a salt of a carboxylate, a sulfonate, a phosphonate, or ammonium.

In certain embodiments, G at each occurrence is a moiety that is charged at physiological pH selected from the group consisting of carboxylate, a sulfonate, a phosphonate, and an amine.

In certain embodiments, G at each occurrence is a moiety containing a polyethylene glycol with between 4 and 30 repeating units, or a polyol.

In certain embodiments, the polyol is a sugar.

In certain embodiments, R′ of formula (IIa) or (IId) includes at least one substitutable nitrogen suitable for attachment to a linker.

In certain embodiments, G is selected independently at each occurrence from:

wherein M is hydrogen or a positively charged counterion. In certain embodiments, M is Na + , K + or Li + . In certain embodiments, M is hydrogen. In particular embodiments, G is SO 3 H.

In certain embodiments, G is selected independently at each occurrence from:

wherein M is hydrogen or a positively charged counterion. In certain embodiments, M is hydrogen.

In particular embodiments, G is SO 3 H.

In certain embodiments, R′ is selected from:

or a salt thereof. When Bcl-xL inhibitors of this embodiment are included in an ADC, the linker of the ADC is linked to the nitrogen atom of an available primary or secondary amine group.

In certain embodiments, R′ is selected from:

or a salt thereof. When Bcl-xL inhibitors of this embodiment are included in an ADC, the linker of the ADC is linked to the nitrogen atom of an available primary or secondary amine group.

In certain embodiments, R′ is selected from

›DETAILED DESCRIPTION OF THE INVENTION · 28 of 40

In certain embodiments, Ar 1 of formulae (IIa)-(IId) is selected from

and is optionally substituted with one or more substituents independently selected from halo, cyano, methyl, and halomethyl. In particular embodiments, Ar 1 is

In certain embodiments, Ar 2 is

optionally substituted with one or more substituents, wherein the R 12 —Z 2b —, R′—Z 2b —, #—N(R 4 )—R 13 —Z 2b —, or #—R′—Z 2b — substituents are attached to Ar 2 at any Ar 2 atom capable of being substituted.

In certain embodiments, Ar 2 is selected from:

and is optionally substituted with one or more substituents, wherein the R 12 —Z 2b —, R′—Z 2b —, #—N(R 4 )—R 13 —Z 2b —, or #—R′—Z 2b — substituents are attached to Ar 2 at any

Ar 2 atom capable of being substituted. In certain embodiments, Ar 2 is selected from:

and is optionally substituted with one or more substituents, wherein the R 12 —Z 2b —, R′—Z 2b —, #—N(R 4 )—R 13 —Z 2b —, or # R′—Z 2b — substituents are attached to Ar 2 at any Ar 2 atom capable of being substituted. In certain embodiments, Ar 2 is substituted with one or more solubilizing group. In certain embodiments, the each solubilizing group is, independently of the others, selected from a moiety containing a polyol, a polyethylene glycol with between 4 and 30 repeating units, a salt, or a moiety that is charged at physiological pH.

In certain embodiments, Z 1 of formulae (IIa)-(IId) is N.

In certain embodiments, Z 2a of formulae (IIa)-(IId) is O. In certain embodiments, Z 2a of formulae (IIa)-(IId) is CR 6a R 6b . In certain embodiments, Z 2a of formulae (IIa)-(IId) is S. In certain embodiments, Z 2a of formulae (IIa)-(IId) is —NR 6 C(O)—. In particular embodiments, R 6 is hydrogen.

In certain embodiments, Z 2b of formulae (IIa)-(IId) is O. In certain embodiments, Z 2b of formulae (IIa)-(IId) is NH or CH 2 .

In certain embodiments, R 1 of formulae (IIa)-(IId) is selected from methyl and chloro.

In certain embodiments, R 2 of formulae (IIa)-(IId) is selected from hydrogen and methyl. In particular embodiments, R 2 is hydrogen.

In certain embodiments the Bcl-xL inhibitor is a compound of formula (IIa). In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa), the compound has the structural formula (IIa.1),

or salts thereof, wherein:

Ar 1 , Ar 2 , Z 1 , Z 2a , Z 2b , R 1 , R 2 , R 1a , R 11b , R 12 , G and # are defined as above;

Y is optionally substituted C 1 -C 8 alkylene;

r is 0 or 1; and

s is 1, 2 or 3.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.1), r is 0 and s is 1.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.1), r is 0 and s is 2.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.1), r is 1 and s is 2.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.1), Z 2a is selected from O, NH, CH 2 and S. In particular embodiments, Z 2a is O. In certain embodiments, Z 2a of formula (IIa.1) is —CR 6a R 6b —. In certain embodiments, Z 2a of formula (IIa.1) is CH 2 . In certain embodiments, Z 2a of formula (IIa.1) is S. In certain embodiments, Z 2a of formula (IIa.1) is —NR 6 C(O)—.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.1), Y is selected from ethylene, propylene and butylene. In particular embodiments, Y is selected from ethylene and propylene.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.1), G is selected from

wherein M is hydrogen or a positively charged counterion. In particular embodiments, G is

In particular embodiments, G is SO 3 H.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.1), Ar 2 is selected from

wherein the R 12 —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.1), Ar 2 is selected from

wherein the R 12 —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted.

In particular embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.1), Ar 2 is

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.1), Z 2b —R 12 is selected from H, F, CN, OCH 3 , OH, NH 2 , OCH 2 CH 2 OCH 3 , N(CH 3 )C(═O)CH 3 , CH 2 N(CH 3 )C(═O)CH 3 SCH 3 , C(═O)N(CH 3 ) 2 and OCH 2 CH 2 N(CH 3 )(C(═O)CH 3 ). In particular embodiments, Z 2b —R 12 is selected from H, F and CN. In particular embodiments, Z 2b —R 12 is H.

In embodiments where Z 2b —R 12 is substituted with hydroxyl (OH), the oxygen can serve as the point of attachment to a linking group (See Section 4.4.1.1).

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.1), Ar 1 is

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.1), the group

bonded to the adamantane ring is selected from:

In certain embodiments, a compound of formula (IIa.1) may be converted into the compound of formula IIa.1.1, wherein n is selected from 1-3:

In certain embodiments, the compound of formula IIa.1.1 can be converted into a compound of formula IIa.1.2, wherein L represents a linker and LK represents a linkage formed between a reactive functional group on linker L and a complementary functional group on antibody.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa), the compound has the structural formula (IIa.2),

or salts thereof, wherein:

Ar 1 , Ar 2 , Z 1 , Z 2a , Z 2b , R 1 , R 2 , R 11a , R 11b , R 12 and # are defined as above;

U is selected from N, O and CH, with the proviso that when U is O, then V a and R 21a are absent;

›DETAILED DESCRIPTION OF THE INVENTION · 29 of 40

R 20 is selected from H and C 1 -C 4 alkyl;

R 21a and R 21b are each, independently from one another, absent or selected from H, C 1 -C 4 alkyl and G, where G is selected from a polyol, PEG4-30, a salt and a moiety that is charged at physiological pH;

V a and V b are each, independently from one another, absent or selected from a bond, and an optionally substituted alkylene;

R 20 is selected from H and C 1 -C 4 alkyl; and

s is 1, 2 or 3.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.2), s is 2.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.2), Z 2a is selected from O, NH, CH 2 and S. In particular embodiments, Z 2a is O. In certain embodiments, Z 2a of formula (IIa.2) is CR 6a R 6b . In certain embodiments, Z 2a of formula (IIa.2) is CH 2 . In certain embodiments, Z 2a of formula (IIa.2) is S. In certain embodiments, Z 2a of formula (IIa.2) is —NR 6 C(O)—.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.2), U is selected from N and O. In particular embodiments, U is O.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.2), V a is a bond, R 21a is a C 1 -C 4 alkyl group, V b is selected from methylene and ethylene and R21b is G. In particular embodiments, V a is a bond, R 21a is a methyl group and V b is selected from methylene and ethylene and R 21b is G.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.2), V a is selected from methylene and ethylene, R 21a is G, V b is selected from methylene and ethylene and R 21b is G. In particular embodiments, V a is ethylene, R 21a is G, V b is selected from methylene and ethylene and R 21b is G.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.2), G is selected from

wherein M is hydrogen or a positively charged counterion. In particular embodiments, G is

In particular embodiments, G is SO 3 H.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.2), R 20 is selected from hydrogen and a methyl group.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.2), Ar 2 is selected from

wherein the R 2 —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.2), Ar 2 is selected from

R 12 —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted.

In particular embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.2), Ar 2 is

wherein the R 12 —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.2), Z 2b —R 12 is selected from H, F, CN, OCH 3 , OH, NH 2 , OCH 2 CH 2 OCH 3 , N(CH 3 )C(═O)CH 3 , CH 2 N(CH 3 )C(═O)CH 3 SCH 3 , C(═O)N(CH 3 ) 2 and OCH 2 CH 2 N(CH 3 )(C(═O)CH 3 ). In particular embodiments, Z 2b —R 12 is selected from H, F and CN. In particular embodiments, Z 2b —R 12 is H. In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.2), Ar 1 is

In particular embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.2), Ar 2 is

wherein the R 12 —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa), the compound has the structural formula (IIa.3),

or salts thereof, wherein:

Ar 1 , Ar 2 , Z 1 , Z 2a , Z 2b , R 1 , R 2 , R 11a , R 11b , R 12 and # are defined as above; R b is selected from H, C 1 -C 4 alkyl and J b -G or is optionally taken together with an atom of T to form a ring having between 3 and 7 atoms;

J a and J b are each, independently from one another, selected from optionally substituted C 1 -C 8 alkylene and optionally substituted phenylene;

T is selected from optionally substituted C 1 -C 8 alkylene, CH 2 CH 2 OCH 2 CH 2 OCH 2 CH 2 , CH 2 CH 2 OCH 2 CH 2 OCH 2 CH 2 OCH 2 and a polyethylene glycol containing from 4 to 10 ethylene glycol units;

G is selected from a polyol, PEG4-30, a salt and a moiety that is charged at physiological pH; and

s is 1, 2 or 3.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.3), s is 1.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.3), s is 2.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.3), Z 2a is selected from O, CH 2 and S. In particular embodiments, Z 2a is O. In certain embodiments, Z 2a of formula (IIa.3) is CR 6a R 6b . In certain embodiments, Z 2a of formula (IIa.3) is CH 2 . In certain embodiments, Z 2a of formula (IIa.3) is S. In certain embodiments, Z 2a of formula (IIa.3) is —NR 6 C(O)—.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.3), J a is selected from methylene and ethylene and R b is J b -G, wherein J b is methylene or ethylene. In some such embodiments, T is ethylene. In other such embodiments, T is CH 2 CH 2 OCH 2 CH 2 OCH 2 CH 2 . In other such embodiments, T is a polyethylene glycol containing from 4 to 10 ethylene glycol units.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.3), J a is selected from methylene and ethylene and R b is taken together with an atom of T to form a ring having 4-6 ring atoms.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.3), J a is selected from methylene and ethylene and R b is H or alkyl. In some such embodiments, T is ethylene.

In other such embodiments, T is CH 2 CH 2 OCH 2 CH 2 OCH 2 CH 2 .

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.3), G is selected from

wherein M is hydrogen or a positively charged counterion. In particular embodiments, G is

In particular embodiments, G is SO 3 H.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.3), R 20 is selected from hydrogen and a methyl group.

›DETAILED DESCRIPTION OF THE INVENTION · 30 of 40

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.3), Ar 2 is selected from

wherein the R 12 —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted.

In particular embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.3), Ar 2 is

herein the R 12 —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted. In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.3), Ar 2 is selected from

wherein the R 12 —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted. In particular embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.3), Ar 2 is

wherein the R 12 —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.3), Z 2b —R 12 is selected from H, F, CN, OCH 3 , OH, NH 2 , OCH 2 CH 2 OCH 3 , N(CH 3 )C(═O)CH 3 , CH 2 N(CH 3 )C(═O)CH 3 SCH 3 , C(═O)N(CH 3 ) 2 and OCH 2 CH 2 N(CH 3 )(C(═O)CH 3 ). In particular embodiments, Z 2b —R 12 is selected from H, F and CN. In particular embodiments, Z 2b —R 12 is H.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.3), Ar 1 is

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.3), the group

is selected from:

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIa.3), the group

is selected from:

In certain embodiments the Bcl-xL inhibitor is a compound of formula (IIb). In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIb), the compound has the structural formula (IIb. 1),

or salts thereof, wherein:

Ar 1 , Ar 2 , Z 2a , Z 2b , R 1 , R 2 , R 4 , R 11a , R 11b and # are defined as above;

Y is optionally substituted C 1 -C 8 alkylene;

G is selected from a polyol, PEG4-30, a salt and a moiety that is charged at physiological pH;

r is 0 or 1; and

s is 1, 2 or 3.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIb. 1), s is 1.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIb. 1), s is 2. In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIb. 1), s is 3.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIb. 1), Z 2a is selected from O, CH 2 , NH and S. In particular embodiments, Z 2a is O. In certain embodiments, Z 2a of formula (IIb.1) is CR 6a R 6b . In certain embodiments, Z 2a of formula (IIb.1) is CH 2 . In certain embodiments, Z 2a of formula (IIb. 1) is S. In certain embodiments, Z 2a of formula (IIb.1) is —NR 6 C(O)—.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIb. 1), Z 2b is selected from O, CH 2 , NH, NCH 3 and S. In particular embodiments, Z 2b is O. In particular embodiments, Z 2b is NH. In particular embodiments, Z 2b is NCH 3 .

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIb. 1), Y is ethylene and r is 0.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIb. 1), Y is ethylene and r is 1.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIb. 1), R 4 is H or methyl. In particular embodiments, R 4 is methyl. In other embodiments, R 4 is H.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIb. 1), R 4 is taken together with an atom of Y to form a ring having 4-6 ring atoms. In particular embodiments, the ring is a cyclobutane ring. In other embodiments, the ring is a piperazine ring. In other embodiments, the ring is a morpholine ring.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIb. 1), G is selected from

wherein M is hydrogen or a positively charged counterion. In particular embodiments, G is

In other embodiments, G is SO 3 H. In particular embodiments, G is NH 2 . In other embodiments, G is PO 3 H 2 . In particular embodiments, G is NH 2 . In particular embodiments, G is C(O)OH. In particular embodiments, G is polyol.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIb. 1), Ar 2 is selected from

wherein the G-(CH 2 ) s —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted.

In particular embodiments in which the Bcl-xL inhibitor is a compound of formula (IIb. 1), Ar 2 is

wherein the G-(CH 2 ) s —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted. In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIb.1), Ar 2 is selected from

wherein the G-(CH 2 ) s —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted. In particular embodiments in which the Bcl-xL inhibitor is a compound of formula (IIb. 1), Ar 2 is

wherein the G-(CH 2 ) s —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIb. 1), Ar 1 is

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIb. 1), the group

is selected from:

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIb. 1), the group

is selected from:

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIb. 1), the group

is selected from:

In certain embodiments the Bcl-xL inhibitor is a compound of formula (IIc). In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc), the compound has the structural formula (IIc.1)

or salts thereof, wherein:

Ar 1 , Ar 2 , Z 1 , Z 2a , Z 2b , R 1 , R 2 , R 4 , R 11a , R 11b and # are defined as above;

Y a is optionally substituted C 1 -C 8 alkylene;

Y b is optionally substituted C 1 -C 8 alkylene;

R 23 is selected from H and C 1 -C 4 alkyl; and

G is selected from a polyol, PEG4-30, a salt and a moiety that is charged at physiological pH;

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc. 1), Z 2a is selected from O, CH 2 , NH and S. In particular embodiments, Z 2a is O. In certain embodiments, Z 2a of formula (IIc.1) is CR 6a R 6b . In certain embodiments, Z 2a of formula (IIc.1) is S. In certain embodiments, Z 2a of formula (IIc.1) is —NR 6 C(O)—.

›DETAILED DESCRIPTION OF THE INVENTION · 31 of 40

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc. 1), Z 2b is selected from O, CH 2 , NH, NCH 3 and S. In particular embodiments, Z 2b is O. In particular embodiments, Z 2b is NH. In particular embodiments, Z 2b is NCH 3 .

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc. 1), Z 2b is a bond. In some such embodiments Y a is methylene or ethylene.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc. 1), Z 2b is O. In some such embodiments Y a is methylene, ethylene, or propylene.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc. 1), Z 2b is NR 6 , where R 6 is defined as above. In some such embodiments, R 6 is taken together with an atom from Y a to form a cycloalkyl or heterocyclyl ring having between 3 and 7 ring atoms. In some such embodiments, the ring has 5 atoms.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc. 1), Y a is ethylene.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc. 1), Y a is methylene.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc.1), Y a is propylene.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc. 1), R 4 is H or methyl. In particular embodiments, R 4 is H.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc. 1), Y b is ethylene or propylene. In particular embodiments, Y b is ethylene.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc. 1), R 23 is methyl.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc.1), R 23 is H.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc. 1), G is selected from

wherein M is hydrogen or a positively charged counterion. In particular embodiments, G is

In particular embodiments, G is SO 3 H.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc.1), Ar 2 is selected from

wherein the #—N(R 4 )—Y a —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted.

In particular embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc. 1), Ar 2 is

wherein the #—N(R 4 ) Y a —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted. In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc.1), Ar 2 is selected from

#—N(R 4 )—Y a —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted. In particular embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc. 1), Ar 2 is

wherein the #—N(R 4 ) a —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc. 1), Ar 1 is

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc. 1), the group

is selected from:

In other embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc. 1), the group

is selected from:

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc), the compound has the structural formula (IIc.2),

or salts thereof, wherein:

Ar 1 , Ar 2 , Z 1 , Z 2a , Z 2b , R 1 , R 2 , R 4 , R 11a , R 11b and # are defined as above;

Y a is optionally substituted C 1 -C 8 alkylene;

Y b is optionally substituted C 1 -C 8 alkylene;

Y c is optionally substituted C 1 -C 8 alkylene;

R 23 is selected from H and C 1 -C 4 alkyl;

R 25 is Y b -G or is taken together with an atom of Y c to form a ring having 4-6 ring atoms; and G is selected from a polyol, PEG4-30, a salt and a moiety that is charged at physiological pH.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc.2), Z 2a is selected from O, CH 2 , NH and S. In particular embodiments, Z 2a is O. In certain embodiments, Z 2a of formula (IIc.2) is CR 6a R 6b . In certain embodiments, Z 2a of formula (IIc.2) is S. In certain embodiments, Z 2a of formula (IIc.2) is —NR 6 C(O)—.In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc.2), Z 2b is selected from O, CH 2 , NH, NCH 3 and S. In particular embodiments, Z 2b is O. In particular embodiments, Z 2b is NH. In particular embodiments, Z 2b is NCH 3 .

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc.2), Z 2b is a bond. In some such embodiments Y a is methylene or ethylene.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc.2), Z 2b is NR 6 , where R 6 is defined as above. In some such embodiments, R 6 is taken together with an atom from Y a to form a cycloalkyl or heterocyclyl ring having between 3 and 7 ring atoms. In some such embodiments, the ring has 5 atoms.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc.2), Y a is ethylene.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc.2), Y a is methylene.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc.2), R 4 is H or methyl.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc.2), Y b is ethylene or propylene. In particular embodiments, Y b is ethylene.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc.2), Y c is ethylene or propylene. In particular embodiments, Y b is ethylene.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc.2), R 25 is taken together with an atom of Y C to form a ring having 4 or 5 ring atoms.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc.2), R 23 is methyl.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc.2), G is selected from

wherein M is hydrogen or a positively charged counterion. In particular embodiments, G is

In particular embodiments, G is SO 3 H.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc.2), Ar 2 is selected from

›DETAILED DESCRIPTION OF THE INVENTION · 32 of 40

wherein the #—N(R 4 )—Y a —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted.

In particular embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc.2), Ar 2 is

wherein the #—N(R 4 )—Y a —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted. In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc.2), Ar 2 is selected from

wherein the #—N(R 4 )—Y a —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted. In particular embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc.2), Ar 2 is

wherein the #—N(R 4 )—Y a —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc.2), Ar 1 is

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IIc.2), the group

is selected from:

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IId), the compound has the structural formula (IId.1),

or salts thereof, wherein:

Ar 1 , Ar 2 , Z 1 , Z 2a , Z 2b , R 1 , R 2 , R 11a , R 11b and # are defined as above;

Y a is optionally substituted alkylene;

Y b is optionally substituted alkylene;

R 23 is selected from H and C 1 -C 4 alkyl;

G a is selected from a polyol, PEG4-30, a salt and a moiety that is charged at physiological pH;

G b is selected from a polyol, PEG4-30, a salt and a moiety that is charged at physiological pH;

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IId. 1), s is 1.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IId. 1), s is 2.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IId. 1), Z 2a is selected from O, NH, CH 2 and S. In particular embodiments, Z 2a is O. In certain embodiments, Z 2a of formula (IId.1) is CR 6a R 6b . In certain embodiments, Z 2a of formula (IId.1) is S. In certain embodiments, Z 2a of formula (IId. 1) is —NR 6 C(O)—.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IId. 1), Z 2b is selected from O, NH, CH 2 and S. In particular embodiments, Z 2b is O.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IId. 1), Y a is selected from ethylene, propylene and butylene. In particular embodiments, Y is ethylene.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IId. 1), G a is selected from

wherein M is hydrogen or a positively charged counterion. In particular embodiments, G a is

In particular embodiments, G a is SO 3 H. In particular embodiments, G a is CO 2 H.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IId. 1), G b is selected from

wherein M is hydrogen or a positively charged counterion. In particular embodiments, G b is

In particular embodiments, G b is SO 3 H. In particular embodiments, G b is CO 2 H.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IId. 1), R 23 is methyl.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IId. 1), Ar 2 is selected from

wherein the G a -Y a —N(#)—(CH 2 ) s —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted.

In particular embodiments in which the Bcl-xL inhibitor is a compound of formula (IId. 1), Ar 2 is

wherein the G a -Y a —N(#)—(CH 2 ) s —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted. In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IId. 1), Ar 2 is selected from

wherein the G a -Y a —N(#)—(CH 2 ) s —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted. In particular embodiments in which the Bcl-xL inhibitor is a compound of formula (IId. 1), Ar 2 is

wherein the G a -Y a —N(#)—(CH 2 ) s —Z 2b — substituent is attached to Ar 2 at any Ar 2 atom capable of being substituted.

In certain embodiments in which the Bcl-xL inhibitor is a compound of formula (IId. 1), Ar 1 is

In certain embodiments, R 11a and R 11b of formulae (IIa)-(IId) are the same. In a particular embodiment, R 11a and R 11b are each methyl.

In certain embodiments, the compounds of formulae (IIa)-(IId) include one of the following cores (C.1)-(C.21):

Exemplary Bcl-xL inhibitors according to structural formulae (IIa)-(IId) that may be used in the methods described herein in unconjugated form and/or included in the ADCs described herein include the following compounds, and/or salts thereof:

Notably, when the Bcl-xL inhibitor of the present application is in conjugated form, the hydrogen corresponding to the # position of structural formulae (IIa)-(IId) is not present, forming a monoradical. For example, compound W2.01 (Example 1.1) is 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-[1-({3-[2-({2-[2-(carboxymethoxy)ethoxy]ethyl}amino)ethoxy]-5,7-dimethyltricyclo[3.3.1.13,7]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridine-2-carboxylic acid.

When it is in unconjugated form, it has the following structure:

When the same compound is included in the ADCs as shown in structural formula (IIa) or (IIb), the hydrogen corresponding to the # position is not present, forming a monoradical.

In certain embodiments, the Bcl-xL inhibitors according to structural formulae (IIa)-(IId) are selected from the group consisting of W2.01, W2.02, W2.03, W2.04, W2.05, W2.06, W2.07, W2.08, W2.09, W2.10, W2.11, W2.12, W2.13, W2.14, W2.15, W2.16, W2.17, W2.18, W2.19, W2.20, W2.21, W2.22, W2.23, W2.24, W2.25, W2.26, W2.27, W2.28, W2.29, W2.30, W2.31, W2.32, W2.33, W2.34, W2.35, W2.36, W2.37, W2.38, W2.39, W2.40, W2.41, W2.42, W2.43, W2.44, W2.45, W2.46, W2.47, W2.48, W2.49, W2.50, W2.51, W2.52, W2.53, W2.54, W2.55, W2.56, W2.57, W2.58, W2.59, W2.60, W2.61, W2.62, W2.63, W2.64, W2.65, W2.66, W2.67, W2.68, W2.69, W2.70, W2.71, W2.72, W2.73, W2.74, W2.75, W2.76, W2.77, W2.78, W2.79, W2.80, W2.81, W2.82, W2.83, W2.84, W2.85, W2.86, W2.87, W2.88, W2.89, W2.90, and W2.91, or pharmaceutically acceptable salts thereof.

›DETAILED DESCRIPTION OF THE INVENTION · 33 of 40

In certain embodiments, the ADC, or a pharmaceutically acceptable salt thereof, comprises a drug linked to an antibody by way of a linker, wherein the drug is a Bcl-xL inhibitor selected from the group consisting of W2.01, W2.02, W2.03, W2.04, W2.05, W2.06, W2.07, W2.08, W2.09, W2.10, W2.11, W2.12, W2.13, W2.14, W2.15, W2.16, W2.17, W2.18, W2.19, W2.20, W2.21, W2.22, W2.23, W2.24, W2.25, W2.26, W2.27, W2.28, W2.29, W2.30, W2.31, W2.32, W2.33, W2.34, W2.35, W2.36, W2.37, W2.38, W2.39, W2.40, W2.41, W2.42, W2.43, W2.44, W2.45, W2.46, W2.47, W2.48, W2.49, W2.50, W2.51, W2.52, W2.53, W2.54, W2.55, W2.56, W2.57, W2.58, W2.59, W2.60, W2.61, W2.62, W2.63, W2.64, W2.65, W2.66, W2.67, W2.68, W2.69, W2.70, W2.71, W2.72, W2.73, W2.74, W2.75, W2.76, W2.77, W2.78, W2.79, W2.80, W2.81, W2.82, W2.83, W2.84, W2.85, W2.86, W2.87, W2.88, W2.89, W2.90, and W2.91.

In certain embodiments, the ADC, or a pharmaceutically acceptable salt thereof, the Bcl-xL inhibitor is selected from the group consisting of the following compounds modified in that the hydrogen corresponding to the # position of structural formula (IIa), (IIb), (IIc), or (IId) is not present forming a monoradical:

6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-[1-({3-[2-({2-[2-(carboxymethoxy)ethoxy]ethyl}amino)ethoxy]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 2-{[(2-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]amino}ethyl)sulfonyl]amino}-2-deoxy-D-glucopyranose; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(4-{[(3R,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl]methyl}benzyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(3-sulfopropyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[(2,3-dihydroxypropyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 2-({[4-({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]amino}methyl)phenyl]sulfonyl}amino)-2-deoxy-beta-D-glucopyranose; 8-(1,3-benzothiazol-2-ylcarbamoyl)-2-{6-carboxy-5-[1-({3-[2-({2-[1-(beta-D-glucopyranuronosyl)-1H-1,2,3-triazol-4-yl]ethyl}amino)ethoxy]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridin-2-yl}-1,2,3,4-tetrahydroisoquinoline; 3-[1-({3-[2-(2-{[4-(beta-D-allopyranosyloxy)benzyl]amino}ethoxy)ethoxy]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3,5-dimethyl-7-(2-{2-[(2-sulfoethyl)amino]ethoxy}ethoxy)tricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-phosphonoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[methyl(3-sulfo-L-alanyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(3-phosphonopropyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(3-sulfo-L-alanyl)amino]ethoxy tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3,5-dimethyl-7-(2-{2-[(3-phosphonopropyl)amino]ethoxy}ethoxy)tricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; 3-{1-[(3-{2-[L-alpha-aspartyl(methyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 6-{4-[({2-[2-(2-aminoethoxy)ethoxy]ethyl}[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]amino)methyl]benzyl}-2,6-anhydro-L-gulonic acid; 4-({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]amino}methyl)phenyl hexopyranosiduronic acid; 6-[1-(1,3-benzothiazol-2-ylcarbamoyl)-1,2,3,4-tetrahydroquinolin-7-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-phosphonoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[1-(1,3-benzothiazol-2-ylcarbamoyl)-1,2,3,4-tetrahydroquinolin-7-yl]-3-{1-[(3,5-dimethyl-7-{2-[methyl(3-sulfo-L-alanyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-([1,3]thiazolo[5,4-b]pyridin-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-([1,3]thiazolo[4,5-b]pyridin-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 6-[1-(1,3-benzothiazol-2-ylcarbamoyl)-1,2,3,4-tetrahydroquinolin-7-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[(2-carboxyethyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(3-phosphonopropyl)(piperidin-4-yl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 3-{1-[(3-{2-[D-alpha-aspartyl(methyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3-(2-{[1-(carboxymethyl)piperidin-4-yl]amino}ethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; N-[(5S)-5-amino-6-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](methyl)amino}-6-oxohexyl]-N,N-dimethylmethanaminium; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[piperidin-4-yl(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-5-(3-phosphonopropoxy)-3,4-dihydroisoquinolin-2(1H)-yl]-3-[1-({3,5-dimethyl-7-[2-(methylamino)ethoxy]tricyclo[3.3.1.1 3,7 ]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3-(2-{[N-(2-carboxyethyl)-L-alpha-aspartyl]amino}ethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; 3-{1-[(3-{2-[(2-aminoethyl)(2-sulfoethyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 6-[5-(2-aminoethoxy)-8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-[1-({3,5-dimethyl-7-[2-(methylamino)ethoxy]tricyclo[3.3.1.1 3,7 ]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-3-{1-[(3,5-dimethyl-7-{2-[(3-sulfopropyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[(2-carboxyethyl)(piperidin-4-yl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(3-sulfo-L-alanyl)(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[{2-[(2-carboxyethyl)amino]ethyl}(2-sulfoethyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 3-{1-[(3,5-dimethyl-7-{2-[(3-phosphonopropyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-([1,3]thiazolo[4,5-b]pyridin-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 3-{1-[(3,5-dimethyl-7-{2-[(3-phosphonopropyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-([1,3]thiazolo[5,4-b]pyridin-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-5-(carboxymethoxy)-3,4-dihydroisoquinolin-2(1H)-yl]-3-[1-({3,5-dimethyl-7-[2-(methylamino)ethoxy]tricyclo[3.3.1.1 3,7 ]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[(3-carboxypropyl)(piperidin-4-yl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 3-{1-[(3-{2-[L-alpha-aspartyl(2-sulfoethyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[(1,3-dihydroxypropan-2-yl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[5-(2-aminoethoxy)-8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[methyl(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-5-{2-[(2-sulfoethyl)amino]ethoxy}-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[methyl(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl) {2-[(2-sulfoethyl)amino]ethyl}amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-5-{2-[(2-carboxyethyl)amino]ethoxy}-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[methyl(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 3-{1-[(3,5-dimethyl-7-{2-[(3-phosphonopropyl)(piperidin-4-yl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-([1,3]thiazolo[4,5-b]pyridin-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 6-[4-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydro-2H-1,4-benzoxazin-6-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-5-(3-sulfopropoxy)-3,4-dihydroisoquinolin-2(1H)-yl]-3-[1-({3,5-dimethyl-7-[2-(methylamino)ethoxy]tricyclo[3.3.1.1 3,7 ]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridine-2-carboxylic acid; 3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[1-([1,3]thiazolo[4,5-b]pyridin-2-ylcarbamoyl)-1,2,3,4-tetrahydroquinolin-7-yl]pyridine-2-carboxylic acid; 3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-([1,3]thiazolo[4,5-b]pyridin-2-ylcarbamoyl)naphthalen-2-yl]pyridine-2-carboxylic acid; (1ξ)-1-({2-[5-(1-{[3-(2-aminoethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)-6-carboxypyridin-2-yl]-8-(1,3-benzothiazol-2-ylcarbamoyl)-1,2,3,4-tetrahydroisoquinolin-5-yl}methyl)-1,5-anhydro-D-glucitol; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[(3-carboxypropyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-3-{1-[(3,5-dimethyl-7-{2-[(3-phosphonopropyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3-(2-{[4-(beta-D-glucopyranosyloxy)benzyl]amino}ethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; 3-(1-{[3-(2-{[4-(beta-D-allopyranosyloxy)benzyl]amino}ethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 3-{1-[(3-{2-[azetidin-3-yl(2-sulfoethyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 3-{1-[(3-{2-[(3-aminopropyl)(2-sulfoethyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridine-2-carboxylic acid; 6-[1-(1,3-benzothiazol-2-ylcarbamoyl)-1,2,3,4-tetrahydroquinolin-7-yl]-3-{1-[(3-{2-[(2-carboxyethyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[(N 6 ,N 6 -dimethyl-L-lysyl)(methyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 3-{1-[(3-{2-[(3-aminopropyl)(methyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[1-(1,3-benzothiazol-2-ylcarbamoyl)-1,2,3,4-tetrahydroquinolin-7-yl]pyridine-2-carboxylic acid; 3-{1-[(3-{2-[azetidin-3-yl(methyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}-6-[1-(1,3-benzothiazol-2-ylcarbamoyl)-1,2,3,4-tetrahydroquinolin-7-yl]pyridine-2-carboxylic acid; N 6 -(37-oxo-2,5,8,11,14,17,20,23,26,29,32,35-dodecaoxaheptatriacontan-37-yl)-L-lysyl-N-[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]-L-alaninamide; methyl 6-[4-(3-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]amino}propyl)-1H-1,2,3-triazol-1-yl]-6-deoxy-beta-L-glucopyranoside; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-3-{1-[(3-{2-[(2-carboxyethyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[5-(1,3-benzothiazol-2-ylcarbamoyl)quinolin-3-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[4-(1,3-benzothiazol-2-ylcarbamoyl)quinolin-6-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[5-(1,3-benzothiazol-2-ylcarbamoyl)quinolin-3-yl]-3-{1-[(3-{2-[(2-carboxyethyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[1-(1,3-benzothiazol-2-ylcarbamoyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 8-(1,3-benzothiazol-2-ylcarbamoyl)-2-{6-carboxy-5-[1-({3-[2-({3-[1-(beta-D-glucopyranuronosyl)-1H-1,2,3-triazol-4-yl]propyl}amino)ethoxy]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridin-2-yl}-1,2,3,4-tetrahydroisoquinoline; 6-[7-(1,3-benzothiazol-2-ylcarbamoyl)-1H-indol-2-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-6-[3-(methylamino)propyl]-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 5-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]amino}-5-deoxy-D-arabinitol; 1-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]amino}-1,2-dideoxy-D-arabino-hexitol; 6-[4-(1,3-benzothiazol-2-ylcarbamoyl)isoquinolin-6-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3-(2-{[3-hydroxy-2-(hydroxymethyl)propyl]amino}ethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; 1-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]amino}-1,2-dideoxy-D-erythro-pentitol; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3,5-dimethyl-7-(2-{[(2S,3S)-2,3,4-trihydroxybutyl]amino}ethoxy)tricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3-(2-{[(2S,3S,4R,5R,6R)-2, 3,4,5,6,7-hexahydroxyheptyl]amino}ethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[({3-[(1,3-dihydroxypropan-2-yl)amino]propyl}sulfonyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[(3-{[1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl]amino}-3-oxopropyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3-(2-{[(3S)-3,4-dihydroxybutyl]amino}ethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; 4-({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]amino}methyl)phenyl beta-D-glucopyranosiduronic acid; 3-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]amino}propyl beta-D-glucopyranosiduronic acid; 6-[4-(1,3-benzothiazol-2-ylcarbamoyl)-2-oxidoisoquinolin-6-yl]-3-[1-({3,5-dimethyl-7-[2-(methylamino)ethoxy]tricyclo[3.3.1.1 3,7 ]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridine-2-carboxylic acid; 6-{8-[(1,3-benzothiazol-2-yl)carbamoyl]-3,4-dihydroisoquinolin-2(1H)-yl}-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]acetamido}tricyclo[3.3.1.1 3,7 ]decan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3,5-dimethyl-7-({2-[(2-sulfoethyl)amino]ethyl}sulfanyl)tricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; and 6-{8-[(1,3-benzothiazol-2-yl)carbamoyl]-3,4-dihydroisoquinolin-2(1H)-yl}-3-{1-[(3,5-dimethyl-7-{3-[(2-sulfoethyl)amino]propyl}tricyclo[3.3.1.1 3,7 ]decan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid;

›DETAILED DESCRIPTION OF THE INVENTION · 34 of 40

and a pharmaceutically acceptable salt thereof.

The Bcl-xL inhibitors bind to and inhibit anti-apoptotic Bcl-xL proteins, inducing apoptosis. The ability of specific Bcl-xL inhibitors according to structural formulae (IIa)-(IId) to bind to and inhibit Bcl-xL activity may be confirmed in standard binding and activity assays, including, for example, the TR-FRET Bcl-xL binding assays described in Tao et al., 2014, ACS Med. Chem. Lett., 5:1088-1093. A specific TR-FRET Bcl-xL binding assay that can be used to confirm Bcl-xL binding is provided in Example 4, below. Typically, Bcl-xL inhibitors useful as inhibitors per se and in the ADCs described herein will exhibit a K i in the binding assay of Example 5 of less than about 1 nM, but may exhibit a significantly lower K i , for example a K i of less than about 1, 0.1, or even 0.01 nM.

Bcl-xL inhibitory activity may also be confirmed in standard cell-based cytotoxicity assays, such as the FL5.12 cellular and Molt-4 cytotoxicity assays described in Tao et al., 2014, ACS Med. Chem. Lett., 5:1088-1093. A specific Molt-4 cellular cytotoxicity assay that may be used to confirm Bcl-xL inhibitory activity of specific Bcl-xL inhibitors that are able to permeate cell membranes is provided in Examples 5 and 6, below. Typically, such cell-permeable Bcl-xL inhibitors will exhibit an EC 50 of less than about 500 nM in the Molt-4 cytotoxicity assay of Examples 5 and 6, but may exhibit a significantly lower EC 50 , for example an EC 50 of less than about 250, 100, 50, 20, 10 or even 5 nM.

Owing to the presence of solubilizing groups, many of the Bcl-xL inhibitors described herein are expected to exhibit low or very low cell permeability, and therefore will not yield significant activity in certain cellular assays due to the inability of the compound to traverse the cell membrane, including the Molt-4 cellular toxicity assay of Examples 5 and 6. Bcl-xL inhibitory activity of compounds that do not freely traverse cell membranes may be confirmed in cellular assays with permeabilized cells. The process of mitochondrial outer-membrane permeabilization (MOMP) is controlled by the Bcl-2 family proteins. Specifically, MOMP is promoted by the pro-apoptotic Bcl-2 family proteins Bax and Bak which, upon activation oligomerize on the outer mitochondrial membrane and form pores, leading to release of cytochrome c (cyt c). The release of cyt c triggers formulation of the apoptosome which, in turn, results in caspase activation and other events that commit the cell to undergo programmed cell death (see, Goldstein et al., 2005 , Cell Death and Differentiation 12:453-462). The oligomerization action of Bax and Bak is antagonized by the anti-apoptotic Bcl-2 family members, including Bcl-2 and Bcl-xL. Bcl-xL inhibitors, in cells that depend upon Bcl-xL for survival, can cause activation of Bax and/or Bak, MOMP, release of cyt c and downstream events leading to apoptosis. The process of cyt c release can be measured via western blot of both mitochondrial and cytosolic fractions of cells and used as a proxy measurement of apoptosis in cells.

As a means of detecting Bcl-xL inhibitory activity and consequent release of cyt c for Bcl-xL inhibitors with low cell permeability, the cells can be treated with an agent that causes selective pore formation in the plasma, but not mitochondrial, membrane. Specifically, the cholesterol/phospholipid ratio is much higher in the plasma membrane than the mitochondrial membrane. As a result, short incubation with low concentrations of the cholesterol-directed detergent digitonin selectively permeabilizes the plasma membrane without significantly affecting the mitochondrial membrane. This agent forms insoluble complexes with cholesterol leading to the segregation of cholesterol from its normal phospholipid binding sites. This action, in turn, leads to the formation of holes about 40-50 Å wide in the lipid bilayer. Once the plasma membrane is permeabilized, cytosolic components able to pass over digitonin-formed holes can be washed out, including the cytochrome C that was released from mitochondria to cytosol in the apoptotic cells (Campos, 2006 , Cytometry A 69(6):515-523).

Typically, Bcl-xL inhibitors will yield an EC 50 of less than about 10 nM in the Molt-4 cell permeabilized cyt c assay of Examples 5 and 6, although the compounds may exhibit significantly lower EC 50 s, for example, less than about 5, 1, or even 0.5 nM. As demonstrated in Example 6, Bcl-xL inhibitors having low or very low cell permeability that do not exhibit activity in the standard Molt-4 cellular toxicity assay with non-permeablized cells exhibit potent functional activity, as measured by release of cyt c, in cellular cytotoxicity assays with permeabilized cells. In addition to cytochrome c release, mitochondria undergoing apoptosis frequently lose their transmembrane mitochondrial membrane potential (Bouchier-Hayes et al., 2008 , Methods 44(3): 222-228). JC-1 is a cationic carbocyanine dye that accumulates in mitochondria and fluoresces red when mitochondria are healthy and is lost when the mitochondrial membrane is compromised (percentage depolarization; Smiley et al., 1991 , Proc. Natl. Acad. Sci. USA, 88: 3671-3675; Reers et al., 1991 : Biochemistry, 30: 4480-4486). This loss in signal can be detected in permeabilized cells using a fluorimeter (excitation 545 nm and emission of 590 nm) and is therefore fully quantitative, enhancing both reproducibility and throughput. Typically, Bcl-xL inhibitors will yield an EC 50 of less than about 10 nM in the Molt-4 cell permeabilized JC-1 assay of Examples 5 and 6, although the compounds may exhibit significantly lower EC 50 s, for example, less than about 5, 1, 0.5 or even 0.05 nM. As demonstrated in Example 6, Bcl-xL inhibitors having low or very low cell permeability that do not exhibit activity in the standard Molt-4 cellular toxicity assay with non-permeablized cells exhibit potent functional activity, as measured by their loss of transmembrane mitochondrial membrane potential in the JC-1 assay, in cellular cytotoxicity assays with permeabilized cells. Low permeability Bcl-xL inhibitors also exhibit potent activity when administered to cells in the form of ADCs (see, e.g., Example 8).

›DETAILED DESCRIPTION OF THE INVENTION · 35 of 40

Although many of the Bcl-xL inhibitors of structural formulae (IIa)-(IId) selectively or specifically inhibit Bcl-xL over other anti-apoptotic Bcl-2 family proteins, selective and/or specific inhibition of Bcl-xL is not necessary. The Bcl-xL inhibitors and ADCs comprising the compounds may also, in addition to inhibiting Bcl-xL, inhibit one or more other anti-apoptotic Bcl-2 family proteins, such as, for example, Bcl-2. In some embodiments, the Bcl-xL inhibitors and/or ADCs are selective and/or specific for Bcl-xL. By specific or selective is meant that the particular Bcl-xL inhibitor and/or ADC binds or inhibits Bcl-xL to a greater extent than Bcl-2 under equivalent assay conditions. In specific embodiments, the Bcl-xL inhibitors and/or ADCs exhibit in the range of about 10-fold, 100-fold, or even greater specificity or selectivity for Bcl-xL than Bcl-2 in binding assays.

III.A.2. Bcl-xL Linkers

In the ADCs described herein, the Bcl-xL inhibitors are linked to the antibody by way of linkers. The linker linking a Bcl-xL inhibitor to the antibody of an ADC may be short, long, hydrophobic, hydrophilic, flexible or rigid, or may be composed of segments that each independently has one or more of the above-mentioned properties such that the linker may include segments having different properties. The linkers may be polyvalent such that they covalently link more than one Bcl-xL inhibitor to a single site on the antibody, or monovalent such that covalently they link a single Bcl-xL inhibitor to a single site on the antibody.

As will be appreciated by skilled artisans, the linkers link the Bcl-xL inhibitors to the antibody by forming a covalent linkage to the Bcl-xL inhibitor at one location and a covalent linkage to antibody at another. The covalent linkages are formed by reaction between functional groups on the linker and functional groups on the inhibitors and antibody. As used herein, the expression “linker” is intended to include (i) unconjugated forms of the linker that include a functional group capable of covalently linking the linker to a Bcl-xL inhibitor and a functional group capable of covalently linking the linker to an antibody; (ii) partially conjugated forms of the linker that include a functional group capable of covalently linking the linker to an antibody and that is covalently linked to a Bcl-xL inhibitor, or vice versa; and (iii) fully conjugated forms of the linker that is covalently linked to both a Bcl-xL inhibitor and an antibody. In some specific embodiments of intermediate synthons and ADCs described herein, moieties comprising the functional groups on the linker and covalent linkages formed between the linker and antibody are specifically illustrated as R x and LK, respectively.

The linkers are preferably, but need not be, chemically stable to conditions outside the cell, and may be designed to cleave, immolate and/or otherwise specifically degrade inside the cell. Alternatively, linkers that are not designed to specifically cleave or degrade inside the cell may be used. A wide variety of linkers useful for linking drugs to antibodies in the context of ADCs are known in the art. Any of these linkers, as well as other linkers, may be used to link the Bcl-xL inhibitors to the antibody of the ADCs described herein.

Exemplary polyvalent linkers that may be used to link many Bcl-xL inhibitors to an antibody are described, for example, in U.S. Pat. No. 8,399,512; U.S. Published Application No. 2010/0152725; U.S. Pat. Nos. 8,524,214; 8,349,308; U.S. Published Application No. 2013/189218; U.S. Published Application No. 2014/017265; WO 2014/093379; WO 2014/093394; WO 2014/093640, the contents of which are incorporated herein by reference in their entireties. For example, the Fleximer® linker technology developed by Mersana et al. has the potential to enable high-DAR ADCs with good physicochemical properties. As shown below, the Fleximer® linker technology is based on incorporating drug molecules into a solubilizing poly-acetal backbone via a sequence of ester bonds. The methodology renders highly-loaded ADCs (DAR up to 20) whilst maintaining good physicochemical properties. This methodology could be utilized with Bcl-xL inhibitors as shown in the Scheme below.

To utilize the Fleximer® linker technology depicted in the scheme above, an aliphatic alcohol can be present or introduced into the Bcl-xL inhibitor. The alcohol moiety is then conjugated to an alanine moiety, which is then synthetically incorporated into the Fleximer® linker. Liposomal processing of the ADC in vitro releases the parent alcohol-containing drug.

Additional examples of dendritic type linkers can be found in US 2006/116422; US 2005/271615; de Groot et al., (2003) Angew. Chem. Int. Ed. 42:4490-4494; Amir et al., (2003) Angew. Chem. Int. Ed. 42:4494-4499; Shamis et al., (2004) J. Am. Chem. Soc. 126:1726-1731; Sun et al., (2002) Bioorganic & Medicinal Chemistry Letters 12:2213-2215; Sun et al., (2003) Bioorganic & Medicinal Chemistry 11:1761-1768; King et al., (2002) Tetrahedron Letters 43:1987-1990.

Exemplary monovalent linkers that may be used are described, for example, in Nolting, 2013, Antibody-Drug Conjugates, Methods in Molecular Biology 1045:71-100; Kitson et al., 2013 , CROs/CMOs—Chemica Oggi—Chemistry Today 31(4): 30-36; Ducry et al., 2010 , Bioconjugate Chem. 21:5-13; Zhao et al., 2011, J. Med. Chem. 54:3606-3623; U.S. Pat. Nos. 7,223,837; 8,568,728; 8,535,678; and WO2004010957, the content of each of which is incorporated herein by reference in their entireties.

By way of example and not limitation, some cleavable and noncleavable linkers that may be included in the ADCs described herein are described below.

Cleavable Linkers

In certain embodiments, the linker selected is cleavable in vitro and in vivo. Cleavable linkers may include chemically or enzymatically unstable or degradable linkages. Cleavable linkers generally rely on processes inside the cell to liberate the drug, such as reduction in the cytoplasm, exposure to acidic conditions in the lysosome, or cleavage by specific proteases or other enzymes within the cell. Cleavable linkers generally incorporate one or more chemical bonds that are either chemically or enzymatically cleavable while the remainder of the linker is noncleavable.

›DETAILED DESCRIPTION OF THE INVENTION · 36 of 40

In certain embodiments, a linker comprises a chemically labile group such as hydrazone and/or disulfide groups. Linkers comprising chemically labile groups exploit differential properties between the plasma and some cytoplasmic compartments. The intracellular conditions to facilitate drug release for hydrazone containing linkers are the acidic environment of endosomes and lysosomes, while the disulfide containing linkers are reduced in the cytosol, which contains high thiol concentrations, e.g., glutathione. In certain embodiments, the plasma stability of a linker comprising a chemically labile group may be increased by introducing steric hindrance using substituents near the chemically labile group.

Acid-labile groups, such as hydrazone, remain intact during systemic circulation in the blood's neutral pH environment (pH 7.3-7.5) and undergo hydrolysis and release the drug once the ADC is internalized into mildly acidic endosomal (pH 5.0-6.5) and lysosomal (pH 4.5-5.0) compartments of the cell. This pH dependent release mechanism has been associated with nonspecific release of the drug. To increase the stability of the hydrazone group of the linker, the linker may be varied by chemical modification, e.g., substitution, allowing tuning to achieve more efficient release in the lysosome with a minimized loss in circulation.

Hydrazone-containing linkers may contain additional cleavage sites, such as additional acid-labile cleavage sites and/or enzymatically labile cleavage sites. ADCs including exemplary hydrazone-containing linkers include the following structures:

wherein D and Ab represent the drug and Ab, respectively, and n represents the number of drug-linkers linked to the antibody. In certain linkers such as linker (Ig), the linker comprises two cleavable groups—a disulfide and a hydrazone moiety. For such linkers, effective release of the unmodified free drug requires acidic pH or disulfide reduction and acidic pH. Linkers such as (Ih) and (Ii) have been shown to be effective with a single hydrazone cleavage site.

Other acid-labile groups that may be included in linkers include cis-aconityl-containing linkers. cis-Aconityl chemistry uses a carboxylic acid juxtaposed to an amide bond to accelerate amide hydrolysis under acidic conditions.

Cleavable linkers may also include a disulfide group. Disulfides are thermodynamically stable at physiological pH and are designed to release the drug upon internalization inside cells, wherein the cytosol provides a significantly more reducing environment compared to the extracellular environment. Scission of disulfide bonds generally requires the presence of a cytoplasmic thiol cofactor, such as (reduced) glutathione (GSH), such that disulfide-containing linkers are reasonable stable in circulation, selectively releasing the drug in the cytosol. The intracellular enzyme protein disulfide isomerase, or similar enzymes capable of cleaving disulfide bonds, may also contribute to the preferential cleavage of disulfide bonds inside cells. GSH is reported to be present in cells in the concentration range of 0.5-10 mM compared with a significantly lower concentration of GSH or cysteine, the most abundant low-molecular weight thiol, in circulation at approximately 5 μM. Tumor cells, where irregular blood flow leads to a hypoxic state, result in enhanced activity of reductive enzymes and therefore even higher glutathione concentrations. In certain embodiments, the in vivo stability of a disulfide-containing linker may be enhanced by chemical modification of the linker, e.g., use of steric hindrance adjacent to the disulfide bond.

ADCs including exemplary disulfide-containing linkers include the following structures:

wherein D and Ab represent the drug and antibody, respectively, n represents the number of drug-linkers linked to the antibody and R is independently selected at each occurrence from hydrogen or alkyl, for example. In certain embodiments, increasing steric hindrance adjacent to the disulfide bond increases the stability of the linker. Structures such as (Ij) and (Il) show increased in vivo stability when one or more R groups is selected from a lower alkyl such as methyl.

Another type of linker that may be used is a linker that is specifically cleaved by an enzyme. Such linkers are typically peptide-based or include peptidic regions that act as substrates for enzymes. Peptide based linkers tend to be more stable in plasma and extracellular milieu than chemically labile linkers. Peptide bonds generally have good serum stability, as lysosomal proteolytic enzymes have very low activity in blood due to endogenous inhibitors and the unfavorably high pH value of blood compared to lysosomes. Release of a drug from an antibody occurs specifically due to the action of lysosomal proteases, e.g., cathepsin and plasmin. These proteases may be present at elevated levels in certain tumor tissues. In certain embodiments, the linker is cleavable by a lysosomal enzyme. In certain embodiments, the linker is cleavable by a lysosomal enzyme, and the lysosomal enzyme is Cathepsin B. In certain embodiments, the linker is cleavable by a lysosomal enzyme, and the lysosomal enzyme is β-glucuronidase or β-galactosidase. In certain embodiments, the linker is cleavable by a lysosomal enzyme, and the lysosomal enzyme is β-glucuronidase. In certain embodiments, the linker is cleavable by a lysosomal enzyme, and the lysosomal enzyme is β-galactosidase.

Those skilled in the art recognize the importance of cleavable linkers that are stable to plasma, yet are readily cleaved by a lysosomal enzyme. Disclosed herein, in certain embodiments, are linkers, cleavable by the lysosomal enzymes β-glucuronidase or β-galactosidase, that show improved plasma stability and reduced non-specific release of small molecule drug.

In exemplary embodiments, the cleavable peptide is selected from tetrapeptides such as Gly-Phe-Leu-Gly, Ala-Leu-Ala-Leu or dipeptides such as Val-Cit, Val-Ala, and Phe-Lys. In certain embodiments, dipeptides are preferred over longer polypeptides due to hydrophobicity of the longer peptides.

›DETAILED DESCRIPTION OF THE INVENTION · 37 of 40

A variety of dipeptide-based cleavable linkers useful for linking drugs such as doxorubicin, mitomycin, camptothecin, tallysomycin and auristatin/auristatin family members to antibodies have been described (see, Dubowchik et al., 1998 , J. Org. Chem. 67:1866-1872; Dubowchik et al., 1998 , Bioorg. Med. Chem. Lett. 8:3341-3346; Walker et al., 2002 , Bioorg. Med. Chem. Lett. 12:217-219; Walker et al., 2004 , Bioorg. Med. Chem. Lett. 14:4323-4327; and Francisco et al., 2003 , Blood 102:1458-1465, the contents of each of which are incorporated herein by reference). All of these dipeptide linkers, or modified versions of these dipeptide linkers, may be used in the ADCs described herein. Other dipeptide linkers that may be used include those found in ADCs such as Seattle Genetics' Brentuximab Vendotin SGN-35 (Adcetris™), Seattle Genetics SGN-75 (anti-CD-70, MC-monomethyl auristatin F(MMAF), Celldex Therapeutics glembatumumab (CDX-011) (anti-NMB, Val-Cit-monomethyl auristatin E(MMAE), and Cytogen PSMA-ADC (PSMA-ADC-1301) (anti-PSMA, Val-Cit-MMAE).

Enzymatically cleavable linkers may include a self-immolative spacer to spatially separate the drug from the site of enzymatic cleavage. The direct attachment of a drug to a peptide linker can result in proteolytic release of an amino acid adduct of the drug, thereby impairing its activity. The use of a self-immolative spacer allows for the elimination of the fully active, chemically unmodified drug upon amide bond hydrolysis.

One self-immolative spacer is the bifunctional para-aminobenzyl alcohol group, which is linked to the peptide through the amino group, forming an amide bond, while amine containing drugs may be attached through carbamate functionalities to the benzylic hydroxyl group of the linker (to give a p-amidobenzylcarbamate, PABC). The resulting prodrugs are activated upon protease-mediated cleavage, leading to a 1,6-elimination reaction releasing the unmodified drug, carbon dioxide, and remnants of the linker group. The following scheme depicts the fragmentation of p-amidobenzyl carbamate and release of the drug:

wherein X-D represents the unmodified drug. Heterocyclic variants of this self-immolative group have also been described. See U.S. Pat. No. 7,989,434.

In certain embodiments, the enzymatically cleavable linker is a β-glucuronic acid-based linker. Facile release of the drug may be realized through cleavage of the β-glucuronide glycosidic bond by the lysosomal enzyme β-glucuronidase. This enzyme is present abundantly within lysosomes and is overexpressed in some tumor types, while the enzyme activity outside cells is low. β-Glucuronic acid-based linkers may be used to circumvent the tendency of an ADC to undergo aggregation due to the hydrophilic nature of β-glucuronides. In certain embodiments, β-glucuronic acid-based linkers are preferred as linkers for ADCs linked to hydrophobic drugs. The following scheme depicts the release of the drug from and ADC containing a β-glucuronic acid-based linker:

A variety of cleavable β-glucuronic acid-based linkers useful for linking drugs such as auristatins, camptothecin and doxorubicin analogues, CBI minor-groove binders, and psymberin to antibodies have been described (see, Jeffrey et al., 2006 , Bioconjug. Chem. 17:831-840; Jeffrey et al., Bioorg. Med. Chem. Lett. 17:2278-2280; and Jiang et al., 2005 , J. Am. Chem. Soc. 127:11254-11255, the contents of each of which are incorporated herein by reference). All of these β-glucuronic acid-based linkers may be used in the ADCs described herein. In certain embodiments, the enzymatically cleavable linker is a β-galactoside-based linker. β-Galactoside is present abundantly within lysosomes, while the enzyme activity outside cells is low. Additionally, Bcl-xL inhibitors containing a phenol group can be covalently bonded to a linker through the phenolic oxygen. One such linker, described in U.S. Published App. No. 2009/0318668, relies on a methodology in which a diamino-ethane “SpaceLink” is used in conjunction with traditional “PABO”-based self-immolative groups to deliver phenols. The cleavage of the linker is depicted schematically below using a Bcl-xL inhibitor of the disclosure.

Cleavable linkers may include noncleavable portions or segments, and/or cleavable segments or portions may be included in an otherwise non-cleavable linker to render it cleavable. By way of example only, polyethylene glycol (PEG) and related polymers may include cleavable groups in the polymer backbone. For example, a polyethylene glycol or polymer linker may include one or more cleavable groups such as a disulfide, a hydrazone or a dipeptide.

Other degradable linkages that may be included in linkers include ester linkages formed by the reaction of PEG carboxylic acids or activated PEG carboxylic acids with alcohol groups on a biologically active agent, wherein such ester groups generally hydrolyze under physiological conditions to release the biologically active agent. Hydrolytically degradable linkages include, but are not limited to, carbonate linkages; imine linkages resulting from reaction of an amine and an aldehyde; phosphate ester linkages formed by reacting an alcohol with a phosphate group; acetal linkages that are the reaction product of an aldehyde and an alcohol; orthoester linkages that are the reaction product of a formate and an alcohol; and oligonucleotide linkages formed by a phosphoramidite group, including but not limited to, at the end of a polymer, and a 5′ hydroxyl group of an oligonucleotide.

In certain embodiments, the linker comprises an enzymatically cleavable peptide moiety, for example, a linker comprising structural formula (IVa), (IVb), (IVc) or (IVd):

or a pharmaceutically acceptable salt thereof, wherein:

peptide represents a peptide (illustrated N→C, wherein peptide includes the amino and carboxy “termini”) cleavable by a lysosomal enzyme; T represents a polymer comprising one or more ethylene glycol units or an alkylene chain, or combinations thereof; R a is selected from hydrogen, C 16 alkyl, SO 3 H and CH 2 SO 3 H; R y is hydrogen or C 1-4 alkyl-(O), —(C 1-4 alkylene) s -G 1 or C 1-4 alkyl-(N)—[(C 1-4 alkylene)-G 1 ] 2 ; R z is C 1-4 alkyl-(O), —(C 1-4 alkylene) s -G 2 ;

›DETAILED DESCRIPTION OF THE INVENTION · 38 of 40

G 1 is SO 3 H, CO 2 H, PEG 4-32, or sugar moiety; G 2 is SO 3 H, CO 2 H, or PEG 4-32 moiety; r is 0 or 1; s is 0 or 1; p is an integer ranging from 0 to 5; q is 0 or 1; x is 0 or 1; y is 0 or 1; represents the point of attachment of the linker to the Bcl-xL inhibitor; and * represents the point of attachment to the remainder of the linker.

In certain embodiments, the linker comprises an enzymatically cleavable peptide moiety, for example, a linker comprising structural formula (IVa), (IVb), (IVc), or (IVd), or a pharmaceutically acceptable salt thereof.

In certain embodiments, the peptide is selected from a tripeptide or a dipeptide. In particular embodiments, the dipeptide is selected from: Val-Cit; Cit-Val; Ala-Ala; Ala-Cit; Cit-Ala; Asn-Cit; Cit-Asn; Cit-Cit; Val-Glu; Glu-Val; Ser-Cit; Cit-Ser; Lys-Cit; Cit-Lys; Asp-Cit; Cit-Asp; Ala-Val; Val-Ala; Phe-Lys; Lys-Phe; Val-Lys; Lys-Val; Ala-Lys; Lys-Ala; Phe-Cit; Cit-Phe; Leu-Cit; Cit-Leu; Ile-Cit; Cit-Ile; Phe-Arg; Arg-Phe; Cit-Trp; and Trp-Cit; or a pharmaceutically acceptable salt thereof.

Exemplary embodiments of linkers according to structural formula (IVa) that may be included in the ADCs described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody):

Exemplary embodiments of linkers according to structural formula (IVb), (IVc), or (IVd) that may be included in the ADCs described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody):

In certain embodiments, the linker comprises an enzymatically cleavable sugar moiety, for example, a linker comprising structural formula (Va), (Vb), (Vc), (Vd), or (Ve):

or a pharmaceutically acceptable salt thereof, wherein:

q is 0 or 1; r is 0 or 1; X 1 is CH 2 , O or NH; represents the point of attachment of the linker to the drug; and * represents the point of attachment to the remainder of the linker.

Exemplary embodiments of linkers according to structural formula (Va) that may be included in the ADCs described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody):

Exemplary embodiments of linkers according to structural formula (Vb) that may be included in the ADCs described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody):

Exemplary embodiments of linkers according to structural formula (Vc) that may be included in the ADCs described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody):

Exemplary embodiments of linkers according to structural formula (Vd) that may be included in the ADCs described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody):

Exemplary embodiments of linkers according to structural formula (Ve) that may be included in the ADCs described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody):

Non-Cleavable Linkers

Although cleavable linkers may provide certain advantages, the linkers comprising the ADC described herein need not be cleavable. For noncleavable linkers, the drug release does not depend on the differential properties between the plasma and some cytoplasmic compartments. The release of the drug is postulated to occur after internalization of the ADC via antigen-mediated endocytosis and delivery to lysosomal compartment, where the antibody is degraded to the level of amino acids through intracellular proteolytic degradation. This process releases a drug derivative, which is formed by the drug, the linker, and the amino acid residue to which the linker was covalently attached. The amino-acid drug metabolites from conjugates with noncleavable linkers are more hydrophilic and generally less membrane permeable, which leads to less bystander effects and less nonspecific toxicities compared to conjugates with a cleavable linker. In general, ADCs with noncleavable linkers have greater stability in circulation than ADCs with cleavable linkers. Non-cleavable linkers may be alkylene chains, or maybe polymeric in natures, such as, for example, based upon polyalkylene glycol polymers, amide polymers, or may include segments of alkylene chains, polyalkylene glycols and/or amide polymers. In certain embodiments, the linker comprises a polyethylene glycol segment having from 1 to 6 ethylene glycol units.

A variety of non-cleavable linkers used to link drugs to antibodies have been described. (See, Jeffrey et al., 2006 , Bioconjug. Chem. 17; 831-840; Jeffrey et al., 2007 , Bioorg. Med. Chem. Lett. 17:2278-2280; and Jiang et al., 2005 , J. Am. Chem. Soc. 127:11254-11255, the contents of which are incorporated herein by reference). All of these linkers may be included in the ADCs described herein.

In certain embodiments, the linker is non-cleavable in vivo, for example a linker according to structural formula (VIa), (VIb), (VIc) or (VId) (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody:

or a pharmaceutically acceptable salt thereof, wherein:

R a is selected from hydrogen, alkyl, sulfonate and methyl sulfonate; R x is a moiety including a functional group capable of covalently linking the linker to an antibody; and

represents the point of attachment of the linker to the Bcl-xL inhibitor.

Exemplary embodiments of linkers according to structural formula (VIa)-(VId) that may be included in the ADCs described herein include the linkers illustrated below (as illustrated, the linkers

include a group suitable for covalently linking the linker to an antibody, and represents the point of attachment to a Bcl-xL inhibitor):

›DETAILED DESCRIPTION OF THE INVENTION · 39 of 40

Groups Used to Attach Linkers to Anti-B7-H3 Antibodies

Attachment groups can be electrophilic in nature and include: maleimide groups, activated disulfides, active esters such as NHS esters and HOBt esters, haloformates, acid halides, alkyl and benzyl halides such as haloacetamides. As discussed below, there are also emerging technologies related to “self-stabilizing” maleimides and “bridging disulfides” that can be used in accordance with the disclosure.

Loss of the drug-linker from the ADC has been observed as a result of a maleimide exchange process with albumin, cysteine or glutathione (Alley et al., 2008 , Bioconjugate Chem. 19: 759-769). This is particularly prevalent from highly solvent-accessible sites of conjugation while sites that are partially accessible and have a positively charged environment promote maleimide ring hydrolysis (Junutula et al., 2008 , Nat. Biotechnol. 26: 925-932). A recognized solution is to hydrolyze the succinimide formed from conjugation as this is resistant to deconjugation from the antibody, thereby making the ADC stable in serum. It has been reported previously that the succinimide ring will undergo hydrolysis under alkaline conditions (Kalia et al., 2007 , Bioorg. Med. Chem. Lett. 17: 6286-6289). One example of a “self-stabilizing” maleimide group that hydrolyzes spontaneously under antibody conjugation conditions to give an ADC species with improved stability is depicted in the schematic below. See U.S. Published Application No. 2013/0309256, International Application Publication No. WO 2013/173337, Tumey et al., 2014, Bioconjugate Chem. 25: 1871-1880, and Lyon et al., 2014 , Nat. Biotechnol. 32: 1059-1062. Thus, the maleimide attachment group is reacted with a sulfhydryl of an antibody to give an intermediate succinimide ring. The hydrolyzed form of the attachment group is resistant to deconjugation in the presence of plasma proteins.

As shown above, the maleimide ring of a linker may react with an antibody Ab, forming a covalent attachment as either a succinimide (closed form) or succinamide (open form).

Polytherics has disclosed a method for bridging a pair of sulfhydryl groups derived from reduction of a native hinge disulfide bond. See, Badescu et al., 2014 , Bioconjugate Chem. 25:1124-1136. The reaction is depicted in the schematic below. An advantage of this methodology is the ability to synthesize homogenous DAR4 ADCs by full reduction of IgGs (to give 4 pairs of sulfhydryls) followed by reaction with 4 equivalents of the alkylating agent. ADCs containing “bridged disulfides” are also claimed to have increased stability.

Similarly, as depicted below, a maleimide derivative that is capable of bridging a pair of sulfhydryl groups has been developed. See U.S. Published Application No. 2013/0224228.

In certain embodiments the attachment moiety comprises the structural formulae (VIIa), (VIIb), or (VIIc):

or a pharmaceutically acceptable salt thereof, wherein:

R9 is H or —O—(CH 2 CH 2 O) 11 —CH 3 ;

x is 0 or 1;

y is 0 or 1;

G 3 is —CH 2 CH 2 CH 2 SO 3 H or —CH 2 CH 2 O—(CH 2 CH 2 O) 11 —CH 3 ;

R w is —O—CH 2 CH 2 SO 3 H or —NH(CO)—CH 2 CH 2 O—(CH 2 CH 2 O) 12 —CH 3 ; and

* represents the point of attachment to the remainder of the linker.

In certain embodiments, the linker comprises a segment according to structural formulae (VIIIa), (VIIIb), or (VIIIc):

or a hydrolyzed derivative or a pharmaceutically acceptable salt thereof, wherein:

R 9 is H or —O—(CH 2 CH 2 O) 11 —CH 3 ;

x is 0 or 1;

y is 0 or 1;

G 3 is —CH 2 CH 2 CH 2 SO 3 H or —CH 2 CH 2 O—(CH 2 CH 2 O) 11 —CH 3 ;

R w is —O—CH 2 CH 2 SO 3 H or —NH(CO)—CH 2 CH 2 O—(CH 2 CH 2 O) 12 —CH 3 ;

* represents the point of attachment to the remainder of the linker; and

represents the point of attachment of the linker to the antibody.

Exemplary embodiments of linkers according to structural formula (VIIa) and (VIIb) that may be included in the ADCs described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody):

Exemplary embodiments of linkers according to structural formula (VIIc) that may be included in the ADCs described herein include the linkers illustrated below (as illustrated, the linkers include a group suitable for covalently linking the linker to an antibody):

In certain embodiments, L is selected from the group consisting of IVa.1-IVa.8, IVb.1-IVb.19, IVc.1-IVc.7, IVd.1-IVd.4, Va.1-Va.12, Vb.1-Vb.10, Vc. 1-Vc.11, Vd.1-Vd.6, Ve.1-Ve.2, VIa.1, VIc.1-VIc.2, VId.1-VId.4, VIIa.1-VIIa.4, VIIb.1-VIIb.8, VIIc.1-VIIc.6 in either the closed or open form, and a pharmaceutically acceptable salt thereof.

In certain embodiments, L is selected from the group consisting of IVb.2, IVc.5, IVc.6, IVc.7, IVd.4, Vb.9, VIIa.1, VIIa.3, VIIc.1, VIIc.4, and VIIc.5, wherein the maleimide of each linker has reacted with the antibody Ab, forming a covalent attachment as either a succinimide (closed form) or succinamide (open form), and a pharmaceutically acceptable salt thereof.

In certain embodiments, L is selected from the group consisting of IVb.2, IVc.5, IVc.6, IVd.4, VIIa.1, VIIa.3, VIIc.1, VIIc.4, VIIc.5, wherein the maleimide of each linker has reacted with the antibody Ab, forming a covalent attachment as either a succinimide (closed form) or succinamide(open form), and a pharmaceutically acceptable salt thereof.

In certain embodiments, L is selected from the group consisting of IVb.2, VIIa.3, IVc.6, and VIIc.1, wherein is the attachment point to drug D and @ is the attachment point to the LK, wherein when the linker is in the open form as shown below, @ can be either at the α-position or β-position of the carboxylic acid next to it:

Bcl-xL Linker Selection Considerations

As is known by skilled artisans, the linker selected for a particular ADC may be influenced by a variety of factors, including but not limited to, the site of attachment to the antibody (e.g., lys, cys or other amino acid residues), structural constraints of the drug pharmacophore and the lipophilicity of the drug. The specific linker selected for an ADC should seek to balance these different factors for the specific antibody/drug combination. For a review of the factors that are influenced by choice of linkers in ADCs, see Nolting, Chapter 5 “Linker Technology in Antibody-Drug Conjugates,” In: Antibody-Drug Conjugates: Methods in Molecular Biology , vol. 1045, pp. 71-100, Laurent Ducry (Ed.), Springer Science & Business Medica, LLC, 2013.

›DETAILED DESCRIPTION OF THE INVENTION · 40 of 40

For example, ADCs have been observed to effect killing of bystander antigen-negative cells present in the vicinity of the antigen-positive tumor cells. The mechanism of bystander cell killing by ADCs has indicated that metabolic products formed during intracellular processing of the ADCs may play a role. Neutral cytotoxic metabolites generated by metabolism of the ADCs in antigen-positive cells appear to play a role in bystander cell killing while charged metabolites may be prevented from diffusing across the membrane into the medium and therefore cannot affect bystander killing. In certain embodiments, the linker is selected to attenuate the bystander killing effect caused by cellular metabolites of the ADC. In certain embodiments, the linker is selected to increase the bystander killing effect.

The properties of the linker may also impact aggregation of the ADC under conditions of use and/or storage. Typically, ADCs reported in the literature contain no more than 3-4 drug molecules per antibody molecule (see, e.g., Chari, 2008 , Acc Chem Res 41:98-107). Attempts to obtain higher drug-to-antibody ratios (“DAR”) often failed, particularly if both the drug and the linker were hydrophobic, due to aggregation of the ADC (see King et al., 2002 , J Med Chem 45:4336-4343; Hollander et al., 2008 , Bioconjugate Chem 19:358-361; Burke et al., 2009 Bioconjugate Chem 20:1242-1250). In many instances, DARs higher than 3-4 could be beneficial as a means of increasing potency. In instances where the Bcl-xL inhibitor is hydrophobic in nature, it may be desirable to select linkers that are relatively hydrophilic as a means of reducing ADC aggregation, especially in instances where DARS greater than 3-4 are desired. Thus, in certain embodiments, the linker incorporates chemical moieties that reduce aggregation of the ADCs during storage and/or use. A linker may incorporate polar or hydrophilic groups such as charged groups or groups that become charged under physiological pH to reduce the aggregation of the ADCs. For example, a linker may incorporate charged groups such as salts or groups that deprotonate, e.g., carboxylates, or protonate, e.g., amines, at physiological pH.

Exemplary polyvalent linkers that have been reported to yield DARs as high as 20 that may be used to link numerous Bcl-xL inhibitors to an antibody are described in U.S. Pat. No. 8,399,512; U.S. Published Application No. 2010/0152725; U.S. Pat. Nos. 8,524,214; 8,349,308; U.S. Published Application No. 2013/189218; U.S. Published Application No. 2014/017265; WO 2014/093379; WO 2014/093394; WO 2014/093640, the content of which are incorporated herein by reference in their entireties.

In particular embodiments, the aggregation of the ADCs during storage or use is less than about 40% as determined by size-exclusion chromatography (SEC). In particular embodiments, the aggregation of the ADCs during storage or use is less than 35%, such as less than about 30%, such as less than about 25%, such as less than about 20%, such as less than about 15%, such as less than about 10%, such as less than about 5%, such as less than about 4%, or even less, as determined by size-exclusion chromatography (SEC).

III.A.3. Bcl-xL ADC Synthons

Antibody-Drug Conjugate synthons are synthetic intermediates used to form ADCs. The synthons are generally compounds according to structural formula (III):

›D-L-R x (III) · 1 of 16

or a pharmaceutically acceptable salt thereof, wherein D is a Bcl-xL inhibitor as previously described, L is a linker as previously described, and R x is a reactive group suitable for linking the synthon to an antibody.

In specific embodiments, the intermediate synthons are compounds according to structural formulae (IIIa), (IIIb), (IIIc) and (IIId), below, or a pharmaceutically acceptable salt thereof, where the various substituents Ar 1 , Ar 2 , Z 1 , Z 2a , Z 2b , R′, R 1 , R 2 , R 4 , R 11a , R 11b , R 12 and R 13 are as previously defined for structural formulae (IIa), (IIb), (IIc) and (IId), respectively, L is a linker as previously described and R x is a functional group as described above:

To synthesize an ADC, an intermediate synthon according to structural formula (III), or a salt thereof, is contacted with an antibody of interest under conditions in which functional group R x reacts with a “complementary” functional group on the antibody, F x , to form a covalent linkage.

D-L-R x +[F x m Ab→(I)[D-L-LK m Ab (III)

The identities of groups R x and F x will depend upon the chemistry used to link the synthon to the antibody. Generally, the chemistry used should not alter the integrity of the antibody, for example its ability to bind its target. Preferably, the binding properties of the conjugated antibody will closely resemble those of the unconjugated antibody. A variety of chemistries and techniques for conjugating molecules to biological molecules such as antibodies are known in the art and in particular to antibodies, are well-known. See, e.g., Amon et al., “Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy,” in: Monoclonal Antibodies And Cancer Therapy , Reisfeld et al. Eds., Alan R. Liss, Inc., 1985; Hellstrom et al., “Antibodies For Drug Delivery,” in: Controlled Drug Delivery , Robinson et al., Eds., Marcel Dekker, Inc., 2nd Ed. 1987; Thorpe, “Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review,” in: Monoclonal Antibodies ' 84 : Biological And Clinical Applications , Pinchera et al., Eds., 1985; “Analysis, Results, and Future Prospective of the Therapeutic Use of Radiolabeled Antibody In Cancer Therapy,” in: Monoclonal Antibodies For Cancer Detection And Therapy , Baldwin et al., Eds., Academic Press, 1985; Thorpe et al., 1982 , Immunol. Rev. 62:119-58; PCT publication WO 89/12624. Any of these chemistries may be used to link the synthons to an antibody.

Typically, the synthons are linked to the side chains of amino acid residues of the antibody, including, for example, the primary amino group of accessible lysine residues or the sulthydryl group of accessible cysteine residues. Free sulfhydryl groups may be obtained by reducing interchain disulfide bonds. In certain embodiments, LK is a linkage formed with an amino group on the anti-hB7-H3 antibody Ab. In certain embodiments, LK is an amide, thioether, or thiourea. In certain embodiments, LK is an amide or thiourea. In certain embodiments, LK is a linkage formed with a sulfhydryl group on the anti-hB7-H3 antibody Ab. In certain embodiments, LK is a thioether. In certain embodiments, LK is an amide, thioether, or thiourea; and m is an integer ranging from 1 to 8.

A number of functional groups R x and chemistries useful for linking synthons to accessible lysine residues are known, and include by way of example and not limitation NHS-esters and isothiocyanates.

A number of functional groups R x and chemistries useful for linking synthons to accessible free sulfhydryl groups of cysteine residues are known, and include by way of example and not limitation haloacetyls and maleimides.

However, conjugation chemistries are not limited to available side chain groups. Side chains such as amines may be converted to other useful groups, such as hydroxyls, by linking an appropriate small molecule to the amine. This strategy can be used to increase the number of available linking sites on the antibody by conjugating multifunctional small molecules to side chains of accessible amino acid residues of the antibody. Functional groups R suitable for covalently linking the synthons to these “converted” functional groups are then included in the synthons.

The antibody may also be engineered to include amino acid residues for conjugation. An approach for engineering antibodies to include non-genetically encoded amino acid residues useful for conjugating drugs in the context of ADCs is described in Axup et al., 2003 , Proc Natl Acad Sci 109:16101-16106 and Tian et al., 2014 , Proc Natl Acad Sci 111:1776-1771 as are chemistries and functional groups useful for linking synthons to the non-encoded amino acids.

Exemplary synthons useful for making ADCs described herein include, but are not limited to, the following synthons listed below in Table B.

In certain embodiments, the synthon is selected from the group consisting of synthon examples 2.1, 2.2, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 2.10, 2.11, 2.12, 2.13, 2.14, 2.15, 2.16, 2.17, 2.18, 2.19, 2.20, 2.21, 2.22, 2.23, 2.24, 2.25, 2.26, 2.27, 2.28, 2.29, 2.30, 2.31, 2.32, 2.33, 2.34, 2.35, 2.36, 2.37, 2.38, 2.39, 2.40, 2.41, 2.42, 2.43, 2.44, 2.45, 2.46, 2.47, 2.48, 2.49, 2.50, 2.51, 2.52, 2.53, 2.54, 2.55, 2.56, 2.57, 2.58, 2.59, 2.60, 2.61, 2.62, 2.63, 2.64, 2.65, 2.66, 2.67, 2.68, 2.69, 2.77, 2.78, 2.79, 2.80, 2.81, 2.82, 2.83, 2.84, 2.85, 2.86, 2.87, 2.88, 2.89, 2.90, 2.91, 2.92, 2.93, 2.94, 2.95, 2.96, 2.97, 2.98, 2.101, 2.102, 2.103, 2.104, 2.105, 2.106, 2.107, 2.108, 2.109, 2.110, 2.111, 2.112, 2.113, 2.114, 2.115, 2.116, 2.117, 2.118, 2.119, 2.120, 2.121, 2.122, 2.123, 2.124, 2.125, 2.126, 2.127, 2.128, 2.129, 2.130, 2.131, 2.132, 2.133, 2.134, 2.135, 2.136, 2.137, 2.138, 2.139, 2.140, 2.141, 2.142, 2.143, 2.144, 2.145, 2.146, 2.147, 2.148, 2.149, 2.150, 2.151, 2.152, 2.153, 2.154, 2.155, 2.156, 2.157, 2.158, 2.159, 2.160, 2.161, 2.162, 2.163, 2.164, 2.166, 2.167, 2.168, 2.169, 2.170, 2.171, 2.172, 2.173, 2.174, 2.175, and 2.176, or a pharmaceutically acceptable salt thereof. The compound names of these synthon are provided below:

›D-L-R x (III) · 2 of 16

N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](3-sulfopropyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-[4-({[{2-[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethoxy]ethyl}(2-sulfoethyl)carbamoyl]oxy}methyl)phenyl]-N 5 -carbamoyl-L-ornithinamide; methyl 6-[4-(3-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]([4-({N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N 5 -carbamoyl-L-ornithyl}amino)benzyl]oxy}carbonyl)amino}propyl)-1H-1,2,3-triazol-1-yl]-6-deoxy-beta-L-glucopyranoside; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-(4-{[([2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]{3-[1-(beta-D-glucopyranuronosyl)-1H-1,2,3-triazol-4-yl]propyl}carbamoyl)oxy]methyl}phenyl)-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[(2R)-1-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](methyl)amino}-1-oxo-3-sulfopropan-2-yl]carbamoyl}oxy)methyl]phenyl}-L-alaninamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl][4-(beta-D-glucopyranosyloxy)benzyl]carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[4-(beta-D-allopyranosyloxy)benzyl][2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-phosphonoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-phosphonoethyl)carbamoyl}oxy)methyl]phenyl}-L-alaninamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](3-phosphonopropyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[(2R)-1-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]amino}-1-oxo-3-sulfopropan-2-yl]carbamoyl}oxy)methyl]phenyl}-L-alaninamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-[4-({[{2-[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethoxy]ethyl}(3-phosphonopropyl)carbamoyl]oxy}methyl)phenyl]-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-[4-({[{2-[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethoxy]ethyl}(3-phosphonopropyl)carbamoyl]oxy}methyl)phenyl]-L-alaninamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](3-phosphonopropyl)carbamoyl}oxy)methyl]phenyl}-L-alaninamide; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3-(2-{[(2S)-3-carboxy-2-({[(4-{[(2S)-2-{[(2S)-2-{[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]amino}-3-methylbutanoyl]amino}propanoyl]amino}benzyl)oxy]carbonyl}amino)propanoyl](methyl)amino}ethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl][4-(beta-D-glucopyranuronosyloxy)benzyl]carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[1-(1,3-benzothiazol-2-ylcarbamoyl)-1,2,3,4-tetrahydroquinolin-7-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-phosphonoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{2-carboxy-6-[8-([1,3]thiazolo[5,4-b]pyridin-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[(2R)-1-{[2-({3-[(4-{6-[1-(1,3-benzothiazol-2-ylcarbamoyl)-1,2,3,4-tetrahydroquinolin-7-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](methyl)amino}-1-oxo-3-sulfopropan-2-yl]carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[(2R)-1-{[2-({3-[(4-{6-[1-(1,3-benzothiazol-2-ylcarbamoyl)-1,2,3,4-tetrahydroquinolin-7-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](methyl)amino}-1-oxo-3-sulfopropan-2-yl]carbamoyl}oxy)methyl]phenyl}-L-alaninamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N 5 -carbamoyl-N-{4-[({[2-({3-[(4-{2-carboxy-6-[8-([1,3]thiazolo[4,5-b]pyridin-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[1-(1,3-benzothiazol-2-ylcarbamoyl)-1,2,3,4-tetrahydroquinolin-7-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-carboxyethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-carboxyethyl)carbamoyl}oxy)methyl]phenyl}-L-alaninamide; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3-(2-{[(2R)-3-carboxy-2-({[(4-{[(2S)-2-{[(2S)-2-{[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]amino}-3-methylbutanoyl]amino}propanoyl]amino}benzyl)oxy]carbonyl}amino)propanoyl](methyl)amino}eth oxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl][1-(carboxymethyl)piperidin-4-yl]carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; (S)-6-((2-((3-((4-(6-(8-(benzo[d]thiazol-2-ylcarbamoyl)-3,4-dihydroisoquinin-2(1H)-yl)-2-carboxypyridin-3-yl)-5-methyl-1H-pyrazol-1-yl)methyl)-5,7-dimethyladamantan-1-yl)oxy)ethyl)(methyl)amino)-5-((((4-((S)-2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido)-3-methylbutanamido)-5-ureidopentanamido)benzyl)oxy)carbonyl)amino)-N,N,N-trimethyl-6-oxohexan-1-aminium salt; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-L-alaninamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-[4-({[(4-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)amino}piperidin-1-yl)carbonyl]oxy}methyl)phenyl]-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-5-(3-phosphonopropoxy)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](methyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-[4-({[(4-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](3-phosphonopropyl)amino}piperidin-1-yl)carbonyl]oxy}methyl)phenyl]-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](3-phosphonopropyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-[4-({[(4-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-carboxyethyl)amino}piperidin-1-yl)carbonyl]oxy}methyl)phenyl]-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N 5 -{4-({[(4-{[2-({3-[(4-{2-carboxy-6-[8-([1,3]thiazolo[4,5-b]pyridin-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](3-phosphonopropyl)carbamoyl}oxy)methyl]phenyl}-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{2-carboxy-6-[8-([1,3]thiazolo[5,4-b]pyridin-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](3-phosphonopropyl)carbamoyl}oxy)methyl]phenyl}-L-ornithinamide; N-{6-[(chloroacetyl)amino]hexanoyl}-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-L-alaninamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-5-(carboxymethoxy)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](methyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-[4-({[(2-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)amino}ethyl)(2-carboxyethyl)carbamoyl]oxy}methyl)phenyl]-N 5 -carbamoyl-L-ornithinamide; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-[1-({3-[2-({(2S)-2-[{[(4-{[(2S)-5-(carbamoylamino)-2-{[(2S)-2-{[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]amino}-3-methylbutanoyl]amino}pentanoyl]amino}benzyl)oxy]carbonyl}(2-carboxyethyl)amino]-3-carboxypropanoyl}amino)ethoxy]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3-(2-{[(2S)-2-({[(4-{[(2S)-5-(carbamoylamino)-2-{[(2S)-2-{[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]amino}-3-methylbutanoyl]amino}pentanoyl]amino}benzyl)oxy]carbonyl}amino)-3-carboxypropanoyl](2-sulfoethyl)amino}ethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-[4-({[(4-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](3-carboxypropyl)amino}piperidin-1-yl)carbonyl]oxy}methyl)phenyl]-N 5 -carbamoyl-L-ornithinamide; 4-[(1E)-3-({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-5-(carboxymethoxy)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](methyl)carbamoyl}oxy)prop-1-en-1-yl]-2-({N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-beta-alanyl}amino)phenyl beta-D-glucopyranosiduronic acid; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-[4-({[(2-{[8-(1,3-benzothiazol-2-ylcarbamoyl)-2-(6-carboxy-5-{1-[(3,5-dimethyl-7-{2-[methyl(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridin-2-yl)-1,2,3,4-tetrahydroisoquinolin-5-yl]oxy}ethyl)carbamoyl]oxy}methyl)phenyl]-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-[4-({[(2-{[8-(1,3-benzothiazol-2-ylcarbamoyl)-2-(6-carboxy-5-{1-[(3,5-dimethyl-7-{2-[methyl(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridin-2-yl)-1,2,3,4-tetrahydroisoquinolin-5-yl]oxy}ethyl)(2-sulfoethyl)carbamoyl]oxy}methyl)phenyl]-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-[4-({[(2-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)amino}ethyl)(2-sulfoethyl)carbamoyl]oxy}methyl)phenyl]-N 5 -carbamoyl-L-ornithinamide; N-{6-[(chloroacetyl)amino]hexanoyl}-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[4-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydro-2H-1,4-benzoxazin-6-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-[4-({[(2-{[8-(1,3-benzothiazol-2-ylcarbamoyl)-2-(6-carboxy-5-{1-[(3,5-dimethyl-7-{2-[methyl(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridin-2-yl)-1,2,3,4-tetrahydroisoquinolin-5-yl]oxy}ethyl)(2-carboxyethyl)carbamoyl]oxy}methyl)phenyl]-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-5-(3-sulfopropoxy)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](methyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3-(2-{[(2S)-2-({[(4-{[(2S,3R,4S,5S,6S)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl]oxy}-3-[(3-{[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]amino}propanoyl)amino]benzyl)oxy]carbonyl}amino)-3-sulfopropanoyl](methyl)amino}ethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; 4-[(1E)-3-({[2-({3-[(4-{2-carboxy-6-[8-([1,3]thiazolo[5,4-b]pyridin-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)prop-1-en-1-yl]-2-({N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-beta-alanyl}amino)phenyl beta-D-glucopyranosiduronic acid; 4-[(1E)-3-({[2-({3-[(4-{2-carboxy-6-[8-([1,3]thiazolo[4,5-b]pyridin-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)prop-1-en-1-yl]-2-({N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-beta-alanyl}amino)phenyl beta-D-glucopyranosiduronic acid; 4-[(1E)-3-({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)prop-1-en-1-yl]-2-({N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-beta-alanyl}amino)phenyl beta-D-glucopyranosiduronic acid; 4-[(1E)-3-({[2-({3-[(4-{2-carboxy-6-[8-([1,3]thiazolo[5,4-b]pyridin-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](3-phosphonopropyl)carbamoyl}oxy)prop-1-en-1-yl]-2-({N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-beta-alanyl}amino)phenyl beta-D-glucopyranosiduronic acid; 4-[(1E)-3-({([2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-5-(3-phosphonopropoxy)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](methyl)carbamoyl}oxy)prop-1-en-1-yl]-2-({N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-beta-alanyl}amino)phenyl beta-D-glucopyranosiduronic acid; 4-[(1E)-3-({([2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](3-phosphonopropyl)carbamoyl}oxy)prop-1-en-1-yl]-2-({N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-beta-alanyl}amino)phenyl beta-D-glucopyranosiduronic acid; 4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]-3-[2-(2-{[3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoyl]amino}ethoxy)ethoxy]phenyl beta-D-glucopyranosiduronic acid; 4-[(1E)-3-({[2-({3-[(4-{2-carboxy-6-[8-([1,3]thiazolo[4,5-b]pyridin-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](3-phosphonopropyl)carbamoyl}oxy)prop-1-en-1-yl]-2-({N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-beta-alanyl}amino)phenyl beta-D-glucopyranosiduronic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[(2-carboxyethyl)({[(2E)-3-(4-{[(2S,3R,4S,5S,6S)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl]oxy}-3-[(3-([6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]amino}propanoyl)amino]phenyl)prop-2-en-1-yl]oxy}carbonyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[(2-carboxyethyl){[(4-{[(2S,3R,4S,5S,6S)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl]oxy}-2-[2-(2-{[3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoyl]amino}ethoxy)ethoxy]benzyl)oxy]carbonyl}amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; N-[6-(ethenylsulfonyl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; 4-[(1E)-3-{[(4-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](3-phosphonopropyl)amino}piperidin-1-yl)carbonyl]oxy}prop-1-en-1-yl]-2-({N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-beta-alanyl}amino)phenyl beta-D-glucopyranosiduronic acid; 4-[(1E)-3-{[(4-{[2-({3-[(4-{2-carboxy-6-[8-([1,3]thiazolo[4,5-b]pyridin-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]pyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](3-phosphonopropyl)amino}piperidin-1-yl)carbonyl]oxy}prop-1-en-1-yl]-2-({N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-beta-alanyl}amino)phenyl beta-D-glucopyranosiduronic acid; 4-[(1E)-3-({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)prop-1-en-1-yl]-2-({N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-beta-alanyl}amino)phenyl beta-D-glucopyranosiduronic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-[1-({3-[2-({N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-3-sulfo-L-alanyl}amino)ethoxy]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-[1-({3-[2-(2-{[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl](2-sulfoethyl)amino}ethoxy)ethoxy]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3-(2-{[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl](2-sulfoethyl)amino}ethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{[1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-21-oxo-22-(2-sulfoethyl)-3,6,9,12,15,18-hexaoxa-22-azatetracosan-24-yl]oxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{[1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-21-oxo-22-(2-sulfoethyl)-3,6,9,12,15,18,25-heptaoxa-22-azaheptacosan-27-yl]oxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3-(2-{[6-(ethenylsulfonyl)hexanoyl](2-sulfoethyl)amino}ethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[{6-[(chloroacetyl)amino]hexanoyl}(2-sulfoethyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](3-carboxypropyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-{6-[(bromoacetyl)amino]hexanoyl}-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; 4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](3-carboxypropyl)carbamoyl}oxy)methyl]-3-[2-(2-{[3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoyl]amino}ethoxy)ethoxy]phenyl beta-D-glucopyranosiduronic acid; 4-({[(4-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](3-carboxypropyl)amino}piperidin-1-yl)carbonyl]oxy}methyl)-3-[2-(2-{[3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoyl]amino}ethoxy)ethoxy]phenyl beta-D-glucopyranosiduronic acid; 4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](3-sulfopropyl)carbamoyl}oxy)methyl]-3-[2-(2-{[3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoyl]amino}ethoxy)ethoxy]phenyl beta-D-glucopyranosiduronic acid; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-[4-({[(3-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)amino}azetidin-1-yl)carbonyl]oxy}methyl)phenyl]-N 5 -carbamoyl-L-ornithinamide; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{[26-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-8,24-dioxo-3-(2-sulfoethyl)-11,14,17,20-tetraoxa-3,7,23-triazahexacos-1-yl]oxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-[4-({[(3-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)amino}prop yl)carbamoyl]oxy}methyl)phenyl]-N 5 -carbamoyl-L-ornithinamide; N-{6-[(iodoacetyl)amino]hexanoyl}-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-{6-[(ethenylsulfonyl)amino]hexanoyl}-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-{6-[(ethenylsulfonyl)amino]hexanoyl}-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[(3-{[6-(ethenylsulfonyl)hexanoyl]amino}propyl)(2-sulfoethyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; N-[3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[1-(1,3-benzothiazol-2-ylcarbamoyl)-1,2,3,4-tetrahydroquinolin-7-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[(2-carboxyethyl) {[(2-{[(2S,3R,4S,5S,6S)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl]oxy}-4-[2-(2-{[3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoyl]amino}ethoxy)ethoxy]benzyl)oxy]carbonyl}amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-3-sulfo-L-alanyl-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-carboxyethyl)carbamoyl}oxy)methyl]phenyl}-N-carbamoyl-L-ornithinamide; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{[(43S,46S)-43-({[(4-{[(2S)-2-{[(2S)-2-{[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]amino}-3-methylbutanoyl]amino}propanoyl]amino}benzyl)oxy]carbonyl}amino)-46-methyl-37,44,47-trioxo-2,5,8,11,14,17,20,23,26,29,32,35-dodecaoxa-38,45,48-triazapentacontan-50-yl]oxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[1-(1,3-benzothiazol-2-ylcarbamoyl)-1,2,3,4-tetrahydroquinolin-7-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-carboxyethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-carboxyethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-3-{1-[(3-{2-[(2-carboxyethyl) {[(2-{[(2R,3S,4R,5R,6R)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl]oxy}-4-[2-(2-{[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]amino}ethoxy)ethoxy]benzyl)oxy]carbonyl}amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[5-(1,3-benzothiazol-2-ylcarbamoyl)quinolin-3-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoy}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[4-(1,3-benzothiazol-2-ylcarbamoyl)quinolin-6-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-carboxyethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[4-(1,3-benzothiazol-2-ylcarbamoyl)quinolin-6-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[5-(1,3-benzothiazol-2-ylcarbamoyl)quinolin-3-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-carboxyethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[1-(1,3-benzothiazol-2-ylcarbamoyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[7-(1,3-benzothiazol-2-ylcarbamoyl)-1H-indol-2-yl]-2-carboxypyridin-3-yl}-5-methyl-H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-[4-({[{3-[8-(1,3-benzothiazol-2-ylcarbamoyl)-2-(6-carboxy-5-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridin-2-yl)-1,2,3,4-tetrahydroisoquinolin-6-yl]propyl}(methyl)carbamoyl]oxy}methyl)phenyl]-N 5 -carbamoyl-L-ornithinamide; N-(6-{[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl) acetyl]amino}hexanoyl)-L-valyl-N-{4-[(1 [2-(13-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1l-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-(4-[(([2-((3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl][3-(beta-L-glucopyranuronosyloxy)propyl]carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-(4-[(([2-((3-[(4-{6-[4-(1,3-benzothiazol-2-ylcarbamoyl)isoquinolin-6-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-alpha-glutamyl-L-valyl-N-{4-[(1 [2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetyl]-L-alpha-glutamyl-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; 1-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]({[4-({N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-D-valyl-N 5 -carbamoyl-D-ornithyl}amino)benzyl]oxy}carbonyl)amino}-1,2-dideoxy-D-arabino-hexitol; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-(4-[(([2-((3-[(4-{6-[4-(1,3-benzothiazol-2-ylcarbamoyl)-2-oxidoisoquinolin-6-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](methyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-({(3S,5S)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-oxo-5-[(2-sulfoethoxy)methyl]pyrrolidin-1-yl}acetyl)-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-{(2S)-2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3-[4-(2,5,8,11,14,17,20,23,26,29,32-undecaoxatetratriacontan-34-yloxy)phenyl]propanoyl}-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-({(3S,5S)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-oxo-5-[(2-sulfoethoxy)methyl]pyrrolidin-1-yl}acetyl)-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-{(2S)-2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3-[4-(2,5,8,11,14,17,20,23,26,29,32-undecaoxatetratriacontan-34-yloxy)phenyl]propanoyl}-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; (6S)-2,6-anhydro-6-(2-{2-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]-5-({N-[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetyl]-L-valyl-L-alanyl}amino)phenyl ethyl)-L-gulonic acid; 3-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]({[4-(4-{1 [6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]amino}butyl)-2-(beta-D-glucopyranuronosyloxy)benzyl]oxy}carbonyl)amino}propyl beta-D-glucopyranosiduronic acid; N-{[(3S,5S)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-5-(methoxymethyl)-2-oxopyrrolidin-1-yl]acetyl}-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; (6S)-2,6-anhydro-6-(2-{2-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]-5-({N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-L-alanyl}amino)phenyl}ethyl)-L-gulonic acid; 2-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]-5-(4-{[3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoyl]amino}butyl)phenyl beta-D-glucopyranosiduronic acid; 2-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]-5-[4-({(2S)-2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3-[4-(2,5,8,11,14,17,20,23,26,29,32-undecaoxatetratriacontan-34-yloxy)phenyl]propanoyl}amino)butyl]phenyl beta-D-glucopyranosiduronic acid; (6S)-2,6-anhydro-6-(2-{2-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]-5-[(N-{(2S)-2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3-[4-(2,5,8,11,14,17,20,23,26,29,32-undecaoxatetratriacontan-34-yloxy)phenyl]propanoyl}-L-valyl-L-alanyl)amino]phenyl}ethyl)-L-gulonic acid; 6-(8-(benzo[d]thiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl)-3-(1-((3-(2-((((2-(2-((2S,3R,4R,5S,6S)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)ethyl)-4-((S)-2-((S)-2-(2-((3S,5S)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-oxo-5-((2-sulfoethoxy)methyl)pyrrolidin-1-yl)acetamido)-3-methylbutanamido)propanamido)benzyl)oxy)carbonyl)(2-sulfoethyl)amino)ethoxy)-5,7-dimethyladamantan-1-yl)methyl)-5-methyl-1H-pyrazol-4-yl)picolinic acid; 6-(8-(benzo[d]thiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl)-3-(1-((3-(2-((((2-(((2S,3R,4S,5S,6S)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-4-(4-(2-((3S,5S)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-oxo-5-((2-sulfoethoxy)methyl)pyrrolidin-1-yl)acetamido)butyl)benzyl)oxy)carbonyl)(2-sulfoethyl)amino)ethoxy)-5,7-dimethyladamantan-1-yl)methyl)-5-methyl-1H-pyrazol-4-yl)picolinic acid; 2-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]-5-(4-{[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetyl]amino}butyl)phenyl beta-D-glucopyranosiduronic acid; 2-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]-5-(4-{[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetyl]amino}butyl)phenyl beta-D-glucopyranosiduronic acid; 2-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]-5-[4-({(2S)-2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3-[4-(2,5,8,11,14,17,20,23,26,29,32-undecaoxatetratriacontan-34-yloxy)phenyl]propanoyl}amino)butyl]phenyl beta-D-glucopyranosiduronic acid; N-[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]-3-(4-carboxybutyl)phenyl}-L-alaninamide; 2-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]-5-(3-{[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetyl]amino}propyl)phenyl beta-D-glucopyranosiduronic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[({[2-{[(2S,3R,4S,5S,6S)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl]oxy}-4-(4-{[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetyl]amino}butyl)benzyl]oxy}carbonyl)(3-{[1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl]amino}-3-oxopropyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-(8-(benzo[d]thiazol-2-ylcarbamoyl)naphthalen-2-yl)-3-(1-((3-(2-((((2-(((2S,3R,4S,5S,6S)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-4-(4-(2-((3S,5S)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-oxo-5-((2-sulfoethoxy)methyl)pyrrolidin-1-yl)acetamido)butyl)benzyl)oxy)carbonyl)(2-sulfoethyl)amino)ethoxy)-5,7-dimethyladamantan-1-yl)methyl)-5-methyl-1H-pyrazol-4-yl)picolinic acid; 2-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl][3-hydroxy-2-(hydroxymethyl)propyl]carbamoyl}oxy)methyl]-5-(3-{[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetyl]amino}propyl)phenyl beta-D-glucopyranosiduronic acid; N-({(3S,5S)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-oxo-5-[(2-sulfoethoxy)methyl]pyrrolidin-1-yl}acetyl)-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl J}oxy)methyl]-3-(2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50-heptadecaoxatripentacont-52-yn-53-yl)phenyl}-L-alaninamide; N-({(3S,5S)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-oxo-5-[(2-sulfoethoxy)methyl]pyrrolidin-1-yl}acetyl)-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]-3-(2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50-heptadecaoxatripentacontan-53-yl)phenyl}-L-alaninamide; 2-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl][(3S)-3,4-dihydroxybutyl]carbamoyl}oxy)methyl]-5-(3-{[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetyl]amino}propyl)phenyl beta-D-glucopyranosiduronic acid; 1-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]({[4-(4-{[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetyl]amino}butyl)-2-(beta-D-glucopyranuronosyloxy)benzyl]oxy}carbonyl)amino}-1,2-dideoxy-D-arabino-hexitol; 1-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]({[4-(4-{[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetyl]amino}butyl)-2-(beta-D-glucopyranuronosyloxy)benzyl]oxy}carbonyl)amino}-1,2-dideoxy-D-erythro-pentitol; N-[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl J}oxy)methyl]-3-[27-(2,5,8,11,14,17,20,23-octaoxahexacosan-26-yl)-2,5,8,11,14,17,20,23-octaoxa-27-azatriacontan-30-yl]phenyl}-L-alaninamide; (6S)-2,6-anhydro-6-(2-{2-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]-5-({N-[(2S)-3-[3,4-bis(2,5,8,11,14,17,20,23,26,29,32-undecaoxatetratriacontan-34-yloxy)phenyl]-2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoyl]-L-valyl-L-alanyl}amino)phenyl}ethyl)-L-gulonic acid; N-[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetyl]-N-(2,5,8,11,14,17,20,23,26,29,32-undecaoxatetratriacontan-34-yl)-beta-alanyl-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl J}oxy)methyl]-3-(2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50-heptadecaoxatripentacontan-53-yl)phenyl J}-L-alaninamide; N-[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetyl]-N-(2,5,8,11,14,17,20,23,26,29,32-undecaoxatetratriacontan-34-yl)-beta-alanyl-L-valyl-N-{4-[({[2-(3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]-3-[27-(2,5,8,11,14,17,20,23-octaoxahexacosan-26-yl)-2,5,8,11,14,17,20,23-octaoxa-27-azatriacontan-30-yl]phenyl}-L-alaninamide; N-{(3S)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3-[1-(2,5,8,11,14,17,20,23,26,29,32,35-dodecaoxaheptatriacontan-37-yl)-1H-1,2,3-triazol-4-yl]propanoyl}-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-((3R)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3-[1-(2,5,8,11,14,17,20,23,26,29,32,35-dodecaoxaheptatriacontan-37-yl)-1H-1,2,3-triazol-4-yl]propanoyl}-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-[1-({3-[2-({[(2-{2-[(2S,3R,4R,5S,6S)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl]ethyl}-4-{[(2S)-2-{[(2S)-2-{[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetyl]amino}-3-methylbutanoyl]amino}propanoyl]amino}benzyl)oxy]carbonyl}[(3R,4S,5R)-3,4,5,6-tetrahydroxyhexyl]amino)ethoxy]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-[1-({3-[2-({[(2-{2-[(2S,3R,4R,5S,6S)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl]ethyl}-4-{[(2S)-2-({(2S)-2-[({1 (3S,5S)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-oxo-5-[(2-sulfoethoxy)methyl]pyrrolidin-1-yl}acetyl)amino]-3-methylbutanoyl}amino)propanoyl]amino}benzyl)oxy]carbonyl}[(3R,4S,5R)-3,4,5,6-tetrahydroxyhexyl]amino)ethoxy]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridine-2-carboxylic acid; (6S)-2,6-anhydro-6-(2-{2-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]-5-({N-[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetyl]-N-(2,5,8,11,14,17,20,23,26,29,32-undecaoxatetratriacontan-34-yl)-beta-alanyl-L-valyl-L-alanyl}amino)phenyl}ethyl)-L-gulonic acid; (6S)-2,6-anhydro-6-(2-{2-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]-5-[(N-{2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3-[1-(2,5,8,11,14,17,20,23,26,29,32,35-dodecaoxaheptatriacontan-37-yl)-1H-1,2,3-triazol-4-yl]propanoyl}-L-valyl-L-alanyl)amino]phenyl}ethyl)-L-gulonic acid; (6S)-2,6-anhydro-6-(2-{2-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]-5-[(N-{(3S)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3-[1-(2,5,8,11,14,17,20,23,26,29,32,35-dodecaoxaheptatriacontan-37-yl)-1H-1,2,3-triazol-4-yl]propanoyl}-L-valyl-L-alanyl)amino]phenyl}ethyl)-L-gulonic acid; (6S)-2,6-anhydro-6-(2-{2-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]-5-[(N-{(3R)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3-[1-(2,5,8,11,14,17,20,23,26,29,32,35-dodecaoxaheptatriacontan-37-yl)-1H-1,2,3-triazol-4-yl]propanoyl}-L-valyl-L-alanyl)amino]phenyl}ethyl)-L-gulonic acid; (6S)-2,6-anhydro-6-(2-{2-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]-5-[(N-{(3S)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3-[1-(3-sulfopropyl)-1H-1,2,3-triazol-4-yl]propanoyl}-L-valyl-L-alanyl)amino]phenyl}ethyl)-L-gulonic acid; (6S)-2,6-anhydro-6-(2-{2-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]-5-[(N-{(3R)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3-[1-(3-sulfopropyl)-1H-1,2,3-triazol-4-yl]propanoyl}-L-valyl-L-alanyl)amino]phenyl}ethyl)-L-gulonic acid; (6S)-2,6-anhydro-6-(2-{2-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]-5-({N-[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetyl]-N-[2-(2-sulfoethoxy)ethyl]-beta-alanyl-L-valyl-L-alanyl}amino)phenyl}ethyl)-L-gulonic acid; 6-{8-[(1,3-benzothiazol-2-yl)carbamoyl]-3,4-dihydroisoquinolin-2(1H)-yl}-3-[1-({3-[2-({[(2-{2-[(2S,3R,4R,5S,6S)-6-carboxy-3,4,5-trihydroxyoxan-2-yl]ethyl}-4-{[(2S)-2-{[(2S)-2-{[(2S)-2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3-{4-[(2,5,8,11,14,17,20,23,26,29,32-undecaoxatetratriacontan-34-yl)oxy]phenyl}propanoyl]amino}-3-methylbutanoyl]amino}propanoyl]amino}phenyl)methoxy]carbonyl}[(3R,4S,5R)-3,4,5,6-tetrahydroxyhexyl]amino)ethoxy]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]decan-1-yl}methyl)-5-methyl-1H-pyrazol-4-yl]pyridine-2-carboxylic acid; 4-{[({2-[(3-{[4-(6-{8-[(1,3-benzothiazol-2-yl)carbamoyl]-3,4-dihydroisoquinolin-2(1H)-yl}-2-carboxypyridin-3-yl)-5-methyl-1H-pyrazol-1-yl]methyl}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]decan-1-yl)oxy]ethyl}[(3S)-3,4-dihydroxybutyl]carbamoyl)oxy]methyl}-3-(2-{2-[2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido]ethoxy}ethoxy)phenyl beta-D-glucopyranosiduronic acid; 2,6-anhydro-8-[2-({[{2-[(3-{[4-(6-{8-[(1,3-benzothiazol-2-yl)carbamoyl]-3,4-dihydroisoquinolin-2(1H)-yl}-2-carboxypyridin-3-yl)-5-methyl-1H-pyrazol-1-yl]methyl}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]decan-1-yl)oxy]ethyl}(2-sulfoethyl)carbamoyl]oxy}methyl)-5-{[(79S,82S)-74-[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetyl]-82-methyl-77,80,83-trioxo-79-(propan-2-yl)-2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65,68,71-tetracosaoxa-74,78,81-triazatrioctacontan-83-yl]amino}phenyl]-7,8-dideoxy-L-glycero-L-gulo-octonic acid; 6-{8-[(1,3-benzothiazol-2-yl)carbamoyl]-3,4-dihydroisoquinolin-2(1H)-yl}-3-{1-[(3-{2-[{[(4-{[(2S,5S)-2-[3-(carbamoylamino)propyl]-10-[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetyl]-4,7-dioxo-5-(propan-2-yl)-15-sulfo-13-oxa-3,6,10-triazapentadecanan-1-oyl]amino}phenyl)methoxy]carbonyl}(2-sulfoethyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]decan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-(8-(benzo[d]thiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl)-3-(1-((3-(2-((((2-(2-((2S,3R,4R,5S,6S)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)ethyl)-4-((S)-2-((S)-2-(2-((3S,5S)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-oxo-5-((2-sulfoethoxy)methyl)pyrrolidin-1-yl)acetamido)-3-methylbutanamido)propanamido)benzyl)oxy)carbonyl)((S)-3,4-dihydroxybutyl)amino)ethoxy)-5,7-dimethyladamantan-1-yl)methyl)-5-methyl-1H-pyrazol-4-yl)picolinic acid; 2,6-anhydro-8-(2-{[({2-[(3-{[4-(6-{8-[(1,3-benzothiazol-2-yl)carbamoyl]-3,4-dihydroisoquinolin-2(1H)-yl}-2-carboxypyridin-3-yl)-5-methyl-1H-pyrazol-1-yl]methyl}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]decan-1-yl)oxy]ethyl}[(3S)-3,4-dihydroxybutyl]carbamoyl)oxy]methyl}-5-{[(2S)-2-({(2S)-2-[2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido]-3-methylbutanoyl}amino)propanoyl]amino}phenyl)-7,8-dideoxy-L-glycero-L-gulo-octonic acid; 2-{[({2-[(3-{[4-(6-{8-[(1,3-benzothiazol-2-yl)carbamoyl]-3,4-dihydroisoquinolin-2(1H)-yl}-2-carboxypyridin-3-yl)-5-methyl-1H-pyrazol-1-yl]methyl}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]decan-1-yl)oxy]ethyl}[(3S)-3,4-dihydroxybutyl]carbamoyl)oxy]methyl}-5-{4-[2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido]butyl}phenyl beta-D-glucopyranosiduronic acid; 6-{8-[(1,3-benzothiazol-2-yl)carbamoyl]-3,4-dihydroisoquinolin-2(1H)-yl}-3-{1-[(3-{2-[{[(4-{[(2S)-5-(carbamoylamino)-2-{[(2S)-2-{[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]amino}-3-methylbutanoyl]amino}pentanoyl]amino}phenyl)methoxy]carbonyl}(2-sulfoethyl)amino]acetamido}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]decan-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-{4-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}sulfanyl)ethyl](2-sulfoethyl)carbamoyl}oxy)methyl]phenyl}-N 5 -carbamoyl-L-ornithinamide; N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]-L-valyl-N-[4-({[(3-{3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}propyl)(2-sulfoethyl)carbamoyl]oxy}methyl)phenyl]-N 5 -carbamoyl-L-ornithinamide; 2-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl][(3S)-3,4-dihydroxybutyl]carbamoyl}oxy)methyl]-5-{4-[({(3S,5S)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-oxo-5-[(2-sulfoethoxy)methyl]pyrrolidin-1-yl}acetyl)amino]butyl}phenyl beta-D-glucopyranosiduronic acid; 2,6-anhydro-8-[2-([{2-[(3-{[4-(6-{8-[(1,3-benzothiazol-2-yl)carbamoyl]-3,4-dihydroisoquinolin-2(1H)-yl}-2-carboxypyridin-3-yl)-5-methyl-1H-pyrazol-1-yl]methyl}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]decan-1-yl)oxy]ethyl}(2-sulfoethyl)carbamoyl]oxy}methyl)-5-{[N-({(3R,5S)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-oxo-5-[(2-sulfoethoxy)methyl]pyrrolidin-1-yl}acetyl)-L-valyl-L-alanyl]amino}phenyl]-7,8-dideoxy-L-glycero-L-gulo-octonic acid; 2,6-anhydro-8-{2-(({[{2-[(3-{[4-(6-{8-[(1,3-benzothiazol-2-yl)carbamoyl]-3,4-dihydroisoquinolin-2(1H)-yl}-2-carboxypyridin-3-yl)-5-methyl-1H-pyrazol-1-yl]methyl}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]decan-1-yl)oxy]ethyl}(2-sulfoethyl)carbamoyl]oxy}methyl)-5-[(N-{[(3R,5S)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-oxo-5-(41-oxo-2,5,8,11,14,17,20,23,26,29,32,35,38-tridecaoxa-42-azatritetracontan-43-yl)pyrrolidin-1-yl]acetyl}-L-valyl-L-alanyl)amino]phenyl}-7,8-dideoxy-L-glycero-L-gulo-octonic acid; (6S)-2,6-anhydro-6-(2-{2-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl][(3S)-3,4-dihydroxybutyl]carbamoyl}oxy)methyl]-5-({N-[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetyl]-N-(2,5,8,11,14,17,20,23,26,29,32-undecaoxatetratriacontan-34-yl)-b-alanyl-L-valyl-L-alanyl}amino)phenyl}ethyl)-L-gulonic acid; and (6S)-2,6-anhydro-6-(2-{2-[({[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl][(3S)-3,4-dihydroxybutyl]carbamoyl}oxy)methyl]-5-(({N-[(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetyl]-N-[2-(2-sulfoethoxy)ethyl]-b-alanyl-L-valyl-L-alanyl}amino)phenyl}ethyl)-L-gulonic acid.

›D-L-R x (III) · 3 of 16

In certain embodiments, the ADC, or a pharmaceutically acceptable salt thereof,

D is the Bcl-xL inhibitor selected from the group consisting of the following compounds modified in that the hydrogen corresponding to the # position of structural formula (IIa), (IIb), (IIc), or (IId) is not present, forming a monoradical:

W2.01, W2.02, W2.03, W2.04, W2.05, W2.06, W2.07, W2.08, W2.09, W2.10, W2.11, W2.12, W2.13, W2.14, W2.15, W2.16, W2.17, W2.18, W2.19, W2.20, W2.21, W2.22, W2.23, W2.24, W2.25, W2.26, W2.27, W2.28, W2.29, W2.30, W2.31, W2.32, W2.33, W2.34, W2.35, W2.36, W2.37, W2.38, W2.39, W2.40, W2.41, W2.42, W2.43, W2.44, W2.45, W2.46, W2.47, W2.48, W2.49, W2.50, W2.51, W2.52, W2.53, W2.54, W2.55, W2.56, W2.57, W2.58, W2.59, W2.60, W2.61, W2.62, W2.63, W2.64, W2.65, W2.66, W2.67, W2.68, W2.69, W2.70, W2.71, W2.72, W2.73, W2.74, W2.75, W2.76, W2.77, W2.78, W2.79, W2.80, W2.81, W2.82, W2.83, W2.84, W2.85, W2.86, W2.87, W2.88, W2.89, W2.90, and W2.91, and a pharmaceutically acceptable salt thereof;

L is selected from the group consisting of linkers IVa.1-IVa.8, IVb.1-IVb.19, IVc.1-IVc.7, IVd.1-IVd.4, Va.1-Va.12, Vb. 1-Vb.10, Vc.1-Vc.11, Vd.1-Vd.6, Ve.1-Ve.2, VIa.1, VIc. 1-VIc.2, VId.1-VId.4, VIIa.1-VIIa.4, VIIb.1-VIIb.8, VIIc.1-VIIc.6, wherein each linker has reacted with the antibody, Ab, forming a covalent attachment;

LK is thioether; and

m is an integer ranging from 1 to 8.

In certain embodiments, the ADC, or a pharmaceutically acceptable salt thereof,

6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-{1-[(3-{2-[(2-carboxyethyl)amino]ethoxy}-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)naphthalen-2-yl]-3-{1-[(3,5-dimethyl-7-{2-[(2-sulfoethyl)amino]ethoxy}tricyclo[3.3.1.1 3,7 ]dec-1-yl)methyl]-5-methyl-1H-pyrazol-4-yl}pyridine-2-carboxylic acid; 1-{[2-({3-[(4-{6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-2-carboxypyridin-3-yl}-5-methyl-1H-pyrazol-1-yl)methyl]-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl}oxy)ethyl]amino}-1,2-dideoxy-D-arabino-hexitol; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3-(2-{[3-hydroxy-2-(hydroxymethyl)propyl]amino}ethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid; 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl]-3-(1-{[3-(2-{[(3S)-3,4-dihydroxybutyl]amino}ethoxy)-5,7-dimethyltricyclo[3.3.1.1 3,7 ]dec-1-yl]methyl}-5-methyl-1H-pyrazol-4-yl)pyridine-2-carboxylic acid;

and pharmaceutically acceptable salts thereof;

L is selected from the group consisting of linkers IVb.2, IVc.5, IVc.6, IVc.7, IVd.4, Vb.9, Vc.11, VIIa.1, VIIa.3, VIIc.1, VIIc.4, and VIIc.5 in either closed or open forms and a pharmaceutically acceptable salt thereof;

LK is thioether; and

m is an integer ranging from 2 to 4.

To form an ADC, the maleimide ring of a synthon (for example, the synthons listed in Table B) may react with an antibody Ab, forming a covalent attachment as either a succinimide (closed form) or succinamide(open form). Similarly, other functional groups, e.g. acetyl halide or vinyl sulfone may react with an antibody, Ab, forming a covalent attachment.

In certain embodiments, the ADC, or a pharmaceutically acceptable salt thereof, is selected from the group consisting of AbA-CZ, AbA-TX, AbA-TV, AbA-YY, AbA-AAA, AbA-AAD, AbB-CZ, AbB-TX, AbB-TV, AbB-YY, AbB-AAD, AbG-CZ, AbG-TX, AbG-TV, AbG-YY, AbG-AAA, AbG-AAD, AbK-CZ, AbK-TX, AbK-TV, AbK-YY, AbK-AAA, AbK-AAD, wherein CZ, TX, TV, YY, AAA, and AAD are synthons disclosed in Table B, and wherein the conjugated synthons are either in open or closed form.

In one embodiment, the ADC, or a pharmaceutically acceptable salt thereof, is:

wherein m is 2, Ab is either an anti-hB7-H3 antibody, wherein the anti-hB7-H3 antibody comprises a heavy chain CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 35, a heavy chain CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 34, and a heavy chain CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 33; and a light chain CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 39, a light chain CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 38, and a light chain CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 37; or an anti-hB7-H3 antibody, wherein the anti-hB7H3 antibody comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 147, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 144; or an anti-hB7-H3 antibody, wherein the anti-hB7-H3 antibody comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 168, and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 169.

In one embodiment, the ADC, or a pharmaceutically acceptable salt thereof, is:

wherein m is 2, Ab is either an anti-hB7-H3 antibody, wherein the anti-hB7-H3 antibody comprises a heavy chain CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 12, a heavy chain CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 140, and a heavy chain CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 10; and a light chain CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 15, a light chain CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a light chain CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 136; or an anti-hB7-H3 antibody, wherein the anti-hB7H3 antibody comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 139, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 135; or an anti-hB7-H3 antibody, wherein the anti-hB7-H3 antibody comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 170, and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 171.

›D-L-R x (III) · 4 of 16

In one embodiment, the ADC, or a pharmaceutically acceptable salt thereof, is:

wherein m is 2, Ab is either an anti-hB7-H3 antibody, wherein the anti-hB7-H3 antibody comprises a heavy chain CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 12, a heavy chain CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 140, and a heavy chain CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 10; and a light chain CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 15, a light chain CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a light chain CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 136; or an anti-hB7-H3 antibody, wherein the anti-hB7H3 antibody comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 139, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 135; or an anti-hB7-H3 antibody, wherein the anti-hB7-H3 antibody comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 170, and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 171.

In one embodiment, the ADC, or a pharmaceutically acceptable salt thereof, is:

wherein m is 2, Ab is either an anti-hB7-H3 antibody, wherein the anti-hB7-H3 antibody comprises a heavy chain CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 12, a heavy chain CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 140, and a heavy chain CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 10; and a light chain CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 15, a light chain CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a light chain CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 136; or an anti-hB7-H3 antibody, wherein the anti-hB7H3 antibody comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 139, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 135; or an anti-hB7-H3 antibody, wherein the anti-hB7-H3 antibody comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 170, and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 171.

In one embodiment, the ADC, or a pharmaceutically acceptable salt thereof, is

›D-L-R x (III) · 5 of 16

In one embodiment, the ADC, or a pharmaceutically acceptable salt thereof, is:

wherein m is 2, Ab is either an anti-hB7-H3 antibody, wherein the anti-hB7-H3 antibody comprises a heavy chain CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 12, a heavy chain CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 140, and a heavy chain CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 10; and a light chain CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 15, a light chain CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a light chain CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 136; or an anti-hB7-H3 antibody, wherein the anti-hB7H3 antibody comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 139, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 135; or an anti-hB7-H3 antibody, wherein the anti-hB7-H3 antibody comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 170, and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 171.

In one embodiment, the ADC, or a pharmaceutically acceptable salt thereof, is:

wherein m is 2, Ab is either an anti-hB7-H3 antibody, wherein the anti-hB7-H3 antibody comprises a heavy chain CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 12, a heavy chain CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 140, and a heavy chain CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 10; and a light chain CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 15, a light chain CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a light chain CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 136; or an anti-hB7-H3 antibody, wherein the anti-hB7H3 antibody comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 139, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 135; or an anti-hB7-H3 antibody, wherein the anti-hB7-H3 antibody comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 170, and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 171.

In one embodiment, the ADC, or a pharmaceutically acceptable salt thereof, is:

wherein m is 2, Ab is either an anti-hB7-H3 antibody, wherein the anti-hB7-H3 antibody comprises a heavy chain CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 12, a heavy chain CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 140, and a heavy chain CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 10; and a light chain CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 15, a light chain CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a light chain CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 136; or an anti-hB7-H3 antibody, wherein the anti-hB7H3 antibody comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 139, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 135; or an anti-hB7-H3 antibody, wherein the anti-hB7-H3 antibody comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 170, and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 171.

›D-L-R x (III) · 6 of 16

Bcl-xL inhibitors, including warheads and synthons, and methods of making the same, are described in US 2016/0339117 (AbbVie Inc.), which is incorporated by reference herein.

III.A.4. Methods of Synthesis of Bcl-xL ADCs

The Bcl-xL inhibitors and synthons described herein may be synthesized using standard, known techniques of organic chemistry. General schemes for synthesizing Bcl-xL inhibitors and synthons that may be used as-is or modified to synthesize the full scope of Bcl-xL inhibitors and synthons described herein are provided below. Specific methods for synthesizing exemplary Bcl-xL inhibitors and synthons that may be useful for guidance are provided in the Examples section. ADCs may likewise be prepared by standard methods, such as methods analogous to those described in Hamblett et al., 2004, “Effects of Drug Loading on the Antitumor Activity of a Monoclonal Antibody Drug Conjugate”, Clin. Cancer Res. 10:7063-7070; Doronina et al., 2003, “Development of potent and highly efficacious monoclonal antibody auristatin conjugates for cancer therapy,” Nat. Biotechnol. 21(7):778-784; and Francisco et al., 2003 , Blood 102:1458-1465. For example, ADCs with four drugs per antibody may be prepared by partial reduction of the antibody with an excess of a reducing reagent such as DTT or TCEP at 37° C. for 30 min, then the buffer exchanged by elution through SEPHADEX® G-25 resin with 1 mM DTPA in DPBS. The eluent is diluted with further DPBS, and the thiol concentration of the antibody may be measured using 5,5′-dithiobis(2-nitrobenzoic acid) [Ellman's reagent]. An excess, for example 5-fold, of a linker-drug synthon is added at 4° C. for 1 hr, and the conjugation reaction may be quenched by addition of a substantial excess, for example 20-fold, of cysteine. The resulting ADC mixture may be purified on SEPHADEX G-25 equilibrated in PBS to remove unreacted synthons, desalted if desired, and purified by size-exclusion chromatography. The resulting ADC may then be then sterile filtered, for example, through a 0.2 μm filter, and lyophilized if desired for storage. In certain embodiments, all of the interchain cysteine disulfide bonds are replaced by linker-drug conjugates. One embodiment pertains to a method of making an ADC, comprising contacting a synthon described herein with an antibody under conditions in which the synthon covalently links to the antibody Examples of the foregoing Bcl-xL inhibtors, linkers, and synthons thereof, as well as methods of making the same, can be found in US Patent Publication No. US 2016/0339117, the entire contents of which are incorporated by reference herein.

Specific methods for synthesizing exemplary ADCs that may be used to synthesize the full range of ADCs described herein are provided in the Examples section.

III.A.5. General Methods for Synthesizing Bcl-xL Inhibitors

In the schemes below, the various substituents Ar 1 , Ar 2 , Z 1 , R 4 , R 10 , R 11a and R 11b are as defined in the Detailed Description section.

5.1.1. Synthesis of Compound (6)

The synthesis of an intermediate (6) is described in Scheme 1. Compound (1) can be treated with BH 3 . THF to provide compound (2). The reaction is typically performed at ambient temperature in a solvent, such as, but not limited to, tetrahydrofuran. Compound (3) can be prepared by treating compound (2) with

in the presence of cyanomethylenetributylphosphorane. The reaction is typically performed at an elevated temperature in a solvent such as, but not limited to, toluene. Compound (3) can be treated with ethane-1,2-diol in the presence of a base such as, but not limited to, triethylamine, to provide compound (4). The reaction is typically performed at an elevated temperature, and the reaction may be performed under microwave conditions. Compound (4) can be treated with a strong base, such as, but not limited to, n-butyllithium, followed by the addition of iodomethane, to provide compound (5). The addition and reaction is typically performed in a solvent such as, but not limited to, tetrahydrofuran, at a reduced temperature before warming up to ambient temperature for work up. Compound (5) can be treated with N-iodosuccinimide to provide compound (6). The reaction is typically performed at ambient temperature is a solvent such as, but not limited to, N,N-dimethylformamide.

5.1.2. Synthesis of Compound (12)

The synthesis of intermediate (12) is described in Scheme 2. Compound (3) can be treated with tri-n-butyl-allylstannane in the presence of ZnCl 2 .Et 2 O or N, N′-azoisobutyronitrile (AIBN) to provide compound (10) (Yamamoto et al., 1998 , Heterocycles 47:765-780). The reaction is typically performed at −78° C. in a solvent, such as, but not limited to dichloromethane. Compound (10) can be treated under standard conditions known in the art for hydroboration/oxidation to provide compound (11). For example, treatment of compound (10) with a reagent such as BH 3 .THF in a solvent such as, but not limited to, tetrahydrofuran followed by treatment of the intermediate alkylborane adduct with an oxidant such as, but not limited to, hydrogen peroxide in the presence of a base such as, but not limited to, sodium hydroxide would provide compound (11) (Brown et al., 1968 , J. Am. Chem. Soc. 86:397). Typically the addition of BH 3 .THF is performed at low temperature before warming to ambient temperature, which is followed by the addition of hydrogen peroxide and sodium hydroxide to generate the alcohol product. Compound (12) can be generated according to Scheme 1, as previously described for compound (6).

5.1.3. Synthesis of Compound (15)

The synthesis of intermediate (15) is described in Scheme 3. Compound (3) can be reacted with thiourea in a solvent mixture of acetic acid and 48% aqueous HBr solution at 100° C. to yield an intermediate that can be subsequently treated with sodium hydroxide in a solvent mixture such as, but not limited to, 20% v/v ethanol in water to provide compound (13). Compound (13) can be reacted with 2-chloroethanol in the presence of a base such as, but not limited to, sodium ethoxide to provide compound (14). The reaction is typically performed at ambient or elevated temperatures in a solvent such as, but not limited to, ethanol. Compound (15) can be generated according to Scheme 1, as previously described for compound (6).

›D-L-R x (III) · 7 of 16

5.1.4. Synthesis of Compound (22)

The synthesis of compound (22) is described in Scheme 4. Compound (16) can be reacted with iodomethane in the presence of a base such as, but not limited to, potassium carbonate to provide compound (17). The reaction is typically conducted at ambient or elevated temperature in a solvent such as, but not limited to, acetone or N,N-dimethylformamide. Compound (17) can be reacted under photochemical conditions with tosyl cyanide in the presence of benzophenone to provide compound (18) (see Kamijo et al., 2011 , Org. Lett., 13:5928-5931). The reaction is typically run at ambient temperature in a solvent such as, but not limited to, acetonitrile or benzene using a Riko 100W medium pressure mercury lamp as the light source. Compound (18) can be reacted with lithium hydroxide in a solvent system such as, but not limited to, mixtures of water and tetrahydrofuran or water and methanol to provide compound (19). Compound (19) can be treated with BH 3 .THF to provide compound (20). The reaction is typically performed at ambient temperature in a solvent, such as, but not limited to, tetrahydrofuran. Compound (21) can be prepared by treating compound (20) with

in the presence of cyanomethylenetributylphosphorane. The reaction is typically performed at an elevated temperature in a solvent such as, but not limited to, toluene. Compound (21) can be treated with N-iodosuccinimide to provide compound (22). The reaction is typically performed at ambient temperature is a solvent such as, but not limited to, N,N-dimethylformamide.

5.1.5. Synthesis of Compound (24)

The synthesis of pyrazole compound (24), is described in Scheme 5. Compound (22) can be treated with a reducing agent such as, but not limited to, lithium aluminum hydride in a solvent such as, but not limited to, diethyl ether or tetrahydrofuran to provide compound (23). Typically the reaction is performed at 0° C. before warming to ambient or elevated temperature. Compound (23) can be reacted with di-tert-butyl dicarbonate under standard conditions described herein or in the literature to provide compound (24).

5.1.6. Synthesis of Compound (24a)

The synthesis of intermediate (24a) is described in Scheme 6. Compound (22a) can be hydrolyzed using conditions described in the literature to provide compound (23a). Typically the reaction is run in the presence of potassium hydroxide in a solvent such as, but not limited to, ethylene glycol at elevated temperatures (see Roberts et al., 1994 , J. Org. Chem. 59:6464-6469; Yang et al, 2013 , Org. Lett., 15:690-693). Compound (24a) can be made from compound (23a) by Curtius rearrangement using conditions described in the literature. For example, compound (23a) can be reacted with sodium azide in the presence of tetrabutylammonium bromide, zinc(II) triflate and di-tert-butyl dicarbonate to provide compound (24a) (see Lebel et al., Org. Lett., 2005, 7:4107-4110). Typically the reaction is run at elevated temperatures, preferably from 40-50° C., in a solvent such as, but not limited to, tetrahydrofuran.

5.1.7. Synthesis of Compound (29)

As shown in Scheme 7, compounds of formula (27) can be prepared by reacting compounds of formula (25) with tert-butyl 3-bromo-6-fluoropicolinate (26) in the presence of a base, such as, but not limited to, N,N-diisopropylethylamine, or triethylamine. The reaction is typically performed under an inert atmosphere at an elevated temperature in a solvent, such as, but not limited to, dimethyl sulfoxide. Compounds of formula (27) can be reacted with 4,4,5,5-tetramethyl-1,3,2-dioxaborolane (28), under borylation conditions described herein or in the literature to provide compounds of formula (29).

5.1.8. Synthesis of Compound (38)

Scheme 8 describes a method to make intermediates which contain -Nu (nucleophile) tethered to an adamantane and picolinate protected as a t-butyl ester. Compound (30) can be reacted with compound (31) under Suzuki Coupling conditions described herein or in the literature to provide methyl compound (32). Compound (32) can be treated with a base such as but not limited to triethylamine, followed by methanesulfonyl chloride to provide compound (33). The addition is typically performed at low temperature before warming up to ambient temperature in a solvent, such as, but not limited to, dichloromethane. Compound (33) can be reacted with a nucleophile (Nu) of formula (34) to provide compound (35). Examples of nucleophiles include, but are not limited to, sodium azide, methylamine, ammonia and di-tert-butyl iminodicarbonate. Compound (17) can be reacted with lithium hydroxide to provide compound (36). The reaction is typically performed at ambient temperature in a solvent such as but not limited to tetrahydrofuran, methanol, water, or mixtures thereof. Compound (36) can be reacted with compound (37) under amidation conditions described herein or readily available in the literature to provide compounds of formula (38).

5.1.9. Synthesis of Compounds (42) and (43)

Scheme 9 shows representative methods used to make solubilized Bcl-xL inhibitors. Bcl-xL inhibitors can be synthesized using the general approach of modifying a primary amine with a solubilizing group and then attaching the resulting secondary amine to a linker as described in later schemes. For example, compound (41) can be prepared by reacting compound (39) with compound (40). The reaction is typically performed at ambient temperature in a solvent such as but not limited to N,N-dimethylformamide. Compound (41) can be reacted with trifluoroacetic acid to provide compound (43). The reaction is typically performed at ambient temperature in a solvent such as but not limited to dichloromethane. Another example shown in Scheme 9 is the reaction of compound (39) with diethyl vinylphosphonate, followed by reaction with bromotrimethylsilane and allyltrimethylsilane to provide compound (42). Other examples to introduce solubilizing groups on the Bcl-xL inhibitors described herein include, but are not limited to, reductive amination reactions, alkylations, and amidation reactions.

›D-L-R x (III) · 8 of 16

5.1.10. Synthesis of Compound (47)

Scheme 10 shows introduction of a solubilizing group by amidation reaction. Bcl-xL inhibitors can be synthesized using the general approach of modifying a primary or secondary amine with a solubilizing group and then attaching the resulting amine to a linker as described in later schemes. For example, compound (45) can be treated sequentially with HATU and compound (44), to provide compound (46). Compound (46) can be treated with diethylamine in solvents such as, but not limited to, N,N-dimethylformamide to give compound (47).

5.1.11. Synthesis of Compound (51)

Scheme 11 shows representative methods to make solubilized Bcl-xL inhibitors. Bcl-xL inhibitors can be synthesized using the general approach of modifying a primary amine with a spacer to give a differentially protected diamine. The unprotected secondary amine can be modified with a solubilizing group. Deprotection of a protected amine them reveals a site for linker attachment, as described in later schemes. For example, compound (39) can be reductively alkylated with reagents such as, but not limited to tert-butyl 4-oxopiperidine-1-carboxylate (48), under conditions known in the art, to provide a secondary amine (49). Compound (50) can be prepared by reacting compound (49) with 4-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylbutyl ethenesulfonate (40). The reaction is typically performed at ambient temperature in a solvent such as but not limited to N,N-dimethylformamide. Compound (40) can be reacted with trifluoroacetic acid to provide compound (51). The reaction is typically performed at ambient temperature in a solvent such as but not limited to dichloromethane.

5.1.12. Synthesis of Compound (61)

Scheme 12 describes a method to synthesize solubilized Bcl-xL inhibitors. Compound (52) can be reacted with methanesulfonyl chloride, in the presence of a base, such as, but not limited to, triethylamine, to provide compound (53). The reaction is typically performed at a low temperature in a solvent such as but not limited to dichloromethane. Compound (53) can be treated with ammonia in methanol to provide compound (54). The reaction is typically performed at an elevated temperature, and the reaction may be performed under microwave conditions. Compound (56) can be prepared by reacting compound (55) in the presence of a base such as but not limited to N,N-diisopropylethylamine. The reaction is typically performed at ambient temperature in a solvent such as but not limited to N,N-dimethylformamide. Compound (56) can be treated with di-t-butyldicarbonate and 4-(dimethylamino)pyridine to provide compound (57). The reaction is typically performed at ambient temperature in a solvent such as but not limited to tetrahydrofuran. Compound (59) can be prepared by reacting compound (57) with a boronate ester (or the equivalent boronic acid) of formula (58), under Suzuki Coupling conditions described herein or in the literature. Bis(2,5-dioxopyrrolidin-1-yl) carbonate can be reacted with compound (37), followed by reaction with compound (59), to provide compound (60). The reaction is typically performed at ambient temperature in a solvent such as, but not limited to, acetonitrile. Compound (61) can be prepared by treating compound (60) with trifluoroacetic acid. The reaction is typically performed at ambient temperature in a solvent such as but not limited to dichloromethane.

5.1.13. Synthesis of Compound (70)

Scheme 13 describes the synthesis of 5-hydroxy tetrahydroisoquinoline intermediates. Compound (63) can be prepared by treating compound (62) with N-bromosuccinimide. The reaction is typically performed at ambient temperature is a solvent such as, but not limited to, N,N-dimethylformamide. Compound (63) can be reacted with benzyl bromide in the presence of a base, such as, but not limited to, potassium carbonate, to provide compound (64). The reaction is typically performed at an elevated temperature, in a solvent such as, but not limited to, acetone. Compound (64) can be treated with carbon monoxide and methanol in the presence of a base, such as, but not limited to, triethylamine, and a catalyst, such as, but not limited to, compound (65). The reaction is typically performed at an elevated temperature under an inert atmosphere. Compound (65) can be treated with an acid, such as, but not limited to, hydrochloric acid in dioxane, to provide compound (66). The reaction is typically performed at ambient temperature in a solvent, such as, but not limited to, tetrahydrofuran. Compound (67) can be prepared by reacting compound (66) with tert-butyl 3-bromo-6-fluoropicolinate in the presence of a base, such as, but not limited to, triethylamine. The reaction is typically performed under an inert atmosphere at an elevated temperature in a solvent, such as, but not limited to, dimethyl sulfoxide. Compound (67) can be reacted with a boronic acid of formula (68), wherein Ad is the methyladamantane moiety of the compounds of the disclosure (e.g., the compounds of formulae (IIa)-(IId)), under Suzuki Coupling conditions described herein or in the literature to provide compound (69). Compound (70) can be prepared by reacting compound (69) with hydrogen in the presence of Pd(OH) 2 . The reaction is typically performed at an elevated temperature in a solvent such as, but not limited to tetrahydrofuran.

5.1.14. Synthesis of Compound (75)

Scheme 14 shows representative methods used to make solubilized Bcl-xL inhibitors. Bcl-xL inhibitors can be synthesized using the general approach of modifying an Ar 2 substituent with a solubilizing group and then attaching an amine to a linker as described in later schemes. For example, compound (71) can be reacted with tert-butyl 2-bromoacetate in the presence of a base such as, but not limited to, potassium carbonate in a solvent such as, but not limited, to N,N-dimethylformamide. Compound (72) can be treated with aqueous lithium hydroxide in a solvent such as, but not limited to, methanol, tetrahydrofuran or mixtures thereof to provide compound (73). Compound (74) can be obtained by amidation of compound (73) with compound (37) under conditions previously described. Compound (74) can be treated with acids such as, but not limited to trifluoroacetic acid or HCl, to provide a Bcl-xL inhibitor of the formula (75). The reaction is typically performed at ambient temperature in solvents such as, but not limited to, dichloromethane or 1,4-dioxane.

›D-L-R x (III) · 9 of 16

III.A.6. General Methods for Synthesizing Bcl-xL Synthons

In the schemes below, the various substituents Ar 1 , Ar 2 , Z 1 , Y, G, R 11a and R 11b are as defined in the Detailed Description section.

5.2.1. Synthesis of Compound (89)

As shown in scheme 15, compounds of formula (77), wherein PG is an appropriate base labile protecting group and AA(2) is Cit, Ala, or Lys, can be reacted with 4-(aminophenyl)methanol (78), under amidation conditions described herein or readily available in the literature to provide compound (79). Compound (80) can be prepared by reacting compound (79) with a base such as, but not limited to, diethylamine. The reaction is typically performed at ambient temperature in a solvent such as but not limited to N,N-dimethylformamide. Compound (81), wherein PG is an appropriate base or acid labile protecting group and AA(1) is Val or Phe, can be reacted with compound (80), under amidation conditions described herein or readily available in the literature to provide compound (82). Compound (83) can be prepared by treating compound (82) with diethylamine or trifluoroacetic acid, as appropriate. The reaction is typically performed at ambient temperature in a solvent such as but not limited to dichloromethane. Compound (84), wherein Sp is a spacer, can be reacted with compound (83) to provide compound (85). The reaction is typically performed at ambient temperature in a solvent such as but not limited to N,N-dimethylformamide. Compound (85) can be reacted with bis(4-nitrophenyl) carbonate (86) in the presence of a base such as, but not limited to N,N-diisopropylethylamine, to provide compounds (87). The reaction is typically performed at ambient temperature in a solvent such as but not limited to N,N-dimethylformamide. Compounds (87) can be reacted with compound (88) in the presence of a base such as, but not limited to, N,N-diisopropylethylamine, to provide compound (89). The reaction is typically performed at ambient temperature in a solvent such as, but not limited to, N,N-dimethylformamide.

5.2.2. Synthesis of Compounds (94) and (96)

Scheme 16 describes the installment of alternative mAb-linker attachments to dipeptide Synthons. Compound (88) can be reacted with compound (90) in the presence of a base such as, but not limited to, N,N-diisopropylamine to provide compound (91). The reaction is typically performed at ambient temperature in a solvent such as but not limited to N,N-dimethylformamide. Compound (92) can be prepared by reacting compound (91) with diethylamine. The reaction is typically performed at ambient temperature in a solvent such as but not limited to N,N-dimethylformamide. Compound (93), wherein X 1 is Cl, Br, or I, can be reacted with compound (92), under amidation conditions described herein or readily available in the literature to provide compound (94). Compound (92) can be reacted with compounds of formula (95) under amidation conditions described herein or readily available in the literature to provide compound (96).

5.2.3. Synthesis of Compound (106)

Scheme 17 describes the synthesis of vinyl glucuronide linker intermediates and synthons. (2R,3R,4S,5S,6S)-2-Bromo-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (97) can be treated with silver oxide, followed by 4-bromo-2-nitrophenol (98) to provide (2S,3R,4S,5S,6S)-2-(4-bromo-2-nitrophenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (99). The reaction is typically performed at ambient temperature in a solvent, such as, but not limited to, acetonitrile. (2S,3R,4S,5S,6S)-2-(4-Bromo-2-nitrophenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (99) can be reacted with (E)-tert-butyldimethyl((3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)allyl)oxy)silane (100) in the presence of a base such as, but not limited to, sodium carbonate, and a catalyst such as but not limited to tris(dibenzylideneacetone)dipalladium (Pd 2 (dba) 3 ), to provide (2S,3R,4S,5S,6S)-2-(4-((E)-3-((tert-butyldimethylsilyl)oxy)prop-1-en-1-yl)-2-nitrophenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (101). The reaction is typically performed at an elevated temperature in a solvent, such as, but not limited to, tetrahydrofuran. (2S,3R,4S,5S,6S)-2-(2-amino-4-((E)-3-hydroxyprop-1-en-1-yl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (102) can be prepared by reacting (2S,3R,4S,5S,6S)-2-(4-((E)-3-((tert-butyldimethylsilyl)oxy)prop-1-en-1-yl)-2-nitrophenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (101) with zinc in the presence of an acid such as, but not limited to, hydrochloric acid. The addition is typically performed at low temperature before warming to ambient temperature in a solvent such as, but not limited to, tetrahydrofuran, water, or mixtures thereof. (2S,3R,4S,5S,6S)-2-(2-amino-4-((E)-3-hydroxyprop-1-en-1-yl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (102) can be reacted with (9H-fluoren-9-yl)methyl (3-chloro-3-oxopropyl)carbamate (103), in the presence of a base such as, but not limited to, N,N-diisopropylethylamine, to provide (2S,3R,4S,5S,6S)-2-(2-(3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propanamido)-4-((E)-3-hydroxyprop-1-en-1-yl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (104). The addition is typically performed at low temperature before warming to ambient temperature in a solvent such as, but not limited to, dichloromethane. Compound (88) can be reacted with (2S,3R,4S,5S,6S)-2-(2-(3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propanamido)-4-((E)-3-hydroxyprop-1-en-1-yl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (104) in the presence of a base such as, but not limited to, N-ethyl-N-isopropylpropan-2-amine, followed by work up and reaction with compound (105) in the presence of a base such as, but not limited to, N,N-diisopropylethylamine to provide compound (106). The reactions are typically performed at ambient temperature in a solvent such as, but not limited to N,N-dimethylformamide.

›D-L-R x (III) · 10 of 16

5.2.4. Synthesis of Compound (115)

Scheme 18 describes the synthesis of a representative 2-ether glucuronide linker intermediate and synthon. (2S,3R,4S,5S,6S)-2-Bromo-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (97) can be reacted with 2,4-dihydroxybenzaldehyde (107) in the presence of silver carbonate to provide (2S,3R,4S,5S,6S)-2-(4-formyl-3-hydroxyphenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (108). The reaction is typically performed at an elevated temperature in a solvent, such as, but not limited to, acetonitrile. (2S,3R,4S,5S,6S)-2-(4-Formyl-3-hydroxyphenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (108) can be treated with sodium borohydride to provide (2S,3R,4S,5S,6S)-2-(3-hydroxy-4-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (109). The addition is typically performed at low temperature before warming to ambient temperature in a solvent such as but not limited to tetrahydrofuran, methanol, or mixtures thereof. (2S,3R,4S,5S,6S)-2-(4-(((tert-butyldimethylsilyl)oxy)methyl)-3-hydroxyphenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (110) can be prepared by reacting (2S,3R,4S,5S,6S)-2-(3-hydroxy-4-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (109) with tert-butyldimethylsilyl chloride in the presence of imidazole. The reaction is typically performed at low temperature in a solvent, such as, but not limited to, dichloromethane. (2S,3R,4S,5S,6S)-2-(3-(2-(2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)ethoxy)ethoxy)-4-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (111) can be prepared by reacting (2S,3R,4S,5S,6S)-2-(4-(((tert-butyldimethylsilyl)oxy)methyl)-3-hydroxyphenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (110) with (9H-fluoren-9-yl)methyl (2-(2-hydroxyethoxy)ethyl)carbamate in the presence of triphenylphosphine and a azodicarboxylate such as, but not limited to, di-tert-butyl diazene-1,2-dicarboxylate. The reaction is typically performed at ambient temperature in a solvent such as but not limited to toluene. (2S,3R,4S,5S,6S)-2-(3-(2-(2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)ethoxy)ethoxy)-4-(((tert-butyldimethylsilyl)oxy)methyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (111) can be treated with acetic acid to provide (2S,3R,4S,5S,6S)-2-(3-(2-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)ethoxy)ethoxy)-4-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (112). The reaction is typically performed at ambient temperature in a solvent such as but not limited to water, tetrahydrofuran, or mixtures thereof. (2S,3R,4S,5S,6S)-2-(3-(2-(2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)ethoxy)ethoxy)-4-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (113) can be prepared by reacting (2S,3R,4S,5S,6S)-2-(3-(2-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)ethoxy)ethoxy)-4-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (112) with bis(4-nitrophenyl) carbonate in the presence of a base such as but not limited to N-ethyl-N-isopropylpropan-2-amine. The reaction is typically performed at ambient temperature in a solvent such as but not limited to N,N-dimethylformamide. (2S,3R,4S,5S,6S)-2-(3-(2-(2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)ethoxy)ethoxy)-4-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (113) can be treated with compound (88) in the presence of a base such as but not limited to N-ethyl-N-isopropylpropan-2-amine, followed by treatment with lithium hydroxide to provide a compound (114). The reaction is typically performed at ambient temperature in a solvent such as but not limited to N,N-dimethylformamide, tetrahydrofuran, methanol, or mixtures thereof. Compound (115) can be prepared by reacting compound (114) with compound (84) in the presence of a base such as but not limited to N-ethyl-N-isopropylpropan-2-amine. The reaction is typically performed at ambient temperature in a solvent such as but not limited to N,N-dimethylformamide.

5.2.5. Synthesis of Compound (119)

Scheme 19 describes the introduction of a second solubilizing group to a sugar linker. Compound (116) can be reacted with (R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-sulfopropanoic acid (117), under amidation conditions described herein or readily available in the literature, followed by treatment with a base such as but not limited to diethylamine, to provide compound (118). Compound (118) can be reacted with compound (84), wherein Sp is a spacer, under amidation conditions described herein or readily available in the literature, to provide compound (119).

5.2.6. Synthesis of Compound (129)

Scheme 20 describes the synthesis of 4-ether glucuronide linker intermediates and synthons. 4-(2-(2-Bromoethoxy)ethoxy)-2-hydroxybenzaldehyde (122) can be prepared by reacting 2,4-dihydroxybenzaldehyde (120) with 1-bromo-2-(2-bromoethoxy)ethane (121) in the presence of a base such as, but not limited to, potassium carbonate. The reaction is typically performed at an elevated temperature in a solvent such as but not limited to acetonitrile. 4-(2-(2-Bromoethoxy)ethoxy)-2-hydroxybenzaldehyde (122) can be treated with sodium azide to provide 4-(2-(2-azidoethoxy)ethoxy)-2-hydroxybenzaldehyde (123). The reaction is typically performed at ambient temperature in a solvent such as but not limited to N,N-dimethylformamide. (2S,3R,4S,5S,6S)-2-(5-(2-(2-Azidoethoxy)ethoxy)-2-formylphenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (125) can be prepared by reacting 4-(2-(2-azidoethoxy)ethoxy)-2-hydroxybenzaldehyde (123) with (3R,4S,5S,6S)-2-bromo-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (124) in the presence of silver oxide. The reaction is typically performed at ambient temperature in a solvent such as, but not limited to, acetonitrile. Hydrogenation of (2S,3R,4S,5S,6S)-2-(5-(2-(2-azidoethoxy)ethoxy)-2-formylphenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (125) in the presence of Pd/C will provide (2S,3R,4S,5S,6S)-2-(5-(2-(2-aminoethoxy)ethoxy)-2-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (126). The reaction is typically performed at ambient temperature in a solvent such as, but not limited to, tetrahydrofuran. (2S,3R,4S,5S,6S)-2-(5-(2-(2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)ethoxy)ethoxy)-2-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (127) can be prepared by treating (2S,3R,4S,5S,6S)-2-(5-(2-(2-aminoethoxy)ethoxy)-2-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (126) with (9H-fluoren-9-yl)methyl carbonochloridate in the presence of a base, such as, but not limited to, N-ethyl-N-isopropylpropan-2-amine. The reaction is typically performed at low temperature in a solvent such as, but not limited to, dichloromethane. Compound (88) can be reacted with (2S,3R,4S,5S,6S)-2-(5-(2-(2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)ethoxy)ethoxy)-2-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (127) in the presence of a base, such as, but not limited to, N-ethyl-N-isopropylpropan-2-amine, followed by treatment with lithium hydroxide to provide compound (128). The reaction is typically performed at low temperature in a solvent such as, but not limited to, N,N-dimethylformamide. Compound (129) can be prepared by reacting compound (128) with compound (84) in the presence of a base such as, but not limited to, N-ethyl-N-isopropylpropan-2-amine. The reaction is typically performed at ambient temperature in a solvent such as but not limited to N,N-dimethylformamide.

›D-L-R x (III) · 11 of 16

5.2.7. Synthesis of Compound (139)

Scheme 21 describes the synthesis of carbamate glucuronide intermediates and synthons. 2-Amino-5-(hydroxymethyl)phenol (130) can be treated with sodium hydride and then reacted with 2-(2-azidoethoxy)ethyl 4-methylbenzenesulfonate (131) to provide (4-amino-3-(2-(2-azidoethoxy)ethoxy)phenyl)methanol (132). The reaction is typically performed at an elevated temperature in a solvent such as, but not limited to N,N-dimethylformamide. 2-(2-(2-Azidoethoxy)ethoxy)-4-(((tert-butyldimethylsilyl)oxy)methyl)aniline (133) can be prepared by reacting (4-amino-3-(2-(2-azidoethoxy)ethoxy)phenyl)methanol (132) with tert-butyldimethylchlorosilane in the presence of imidazole. The reaction is typically performed at ambient temperature in a solvent such as, but not limited to tetrahydrofuran. 2-(2-(2-Azidoethoxy)ethoxy)-4-(((tert-butyldimethylsilyl)oxy)methyl)aniline (133) can be treated with phosgene, in the presence of a base such as but not limited to triethylamine, followed by reaction with (3R,4S,5S,6S)-2-hydroxy-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (134) in the presence of a base such as but not limited to triethylamine, to provide 2S,3R,4S,5S,6S)-2-(((2-(2-(2-azidoethoxy)ethoxy)-4-(((tert-butyldimethylsilyl)oxy)methyl)phenyl)carbamoyl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (135). The reaction is typically performed in a solvent such as, but not limited to, toluene, and the additions are typically performed at low temperature, before warming up to ambient temperature after the phosgene addition and heating at an elevated temperature after the (3R,4S,5S,6S)-2-hydroxy-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (134) addition. (2S,3R,4S,5S,6S)-2-(((2-(2-(2-Azidoethoxy)ethoxy)-4-(hydroxymethyl)phenyl)carbamoyl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (136) can be prepared by reacting 2S,3R,4S,5S,6S)-2-(((2-(2-(2-azidoethoxy)ethoxy)-4-(((tert-butyldimethylsilyl)oxy)methyl)phenyl)carbamoyl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (135) with p-toluenesulfonic acid monohydrate. The reaction is typically performed at ambient temperature in a solvent such as, but not limited to methanol. (2S,3R,4S,5S,6S)-2-(((2-(2-(2-Azidoethoxy)ethoxy)-4-(hydroxymethyl)phenyl)carbamoyl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (136) can be reacted with bis(4-nitrophenyl)carbonate in the presence of a base such as, but not limited to, N,N-diisopropylethylamine, to provide (2S,3R,4S,5S,6S)-2-(((2-(2-(2-azidoethoxy)ethoxy)-4-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenyl)carbamoyl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (137). The reaction is typically performed at ambient temperature in a solvent such as, but not limited to, N,N-dimethylformamide. (2S,3R,4S,5S,6S)-2-(((2-(2-(2-Azidoethoxy)ethoxy)-4-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenyl)carbamoyl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (137) can be reacted with compound in the presence of a base such as, but not limited to, N,N-diisopropylethylamine, followed by treatment with aqueous lithium hydroxide, to provide compound (138). The first step is typically conducted at ambient temperature in a solvent such as, but not limited to N,N-dimethylformamide, and the second step is typically conducted at low temperature in a solvent such as but not limited to methanol. Compound (138) can be treated with tris(2-carboxyethyl))phosphine hydrochloride, followed by reaction with compound (84) in the presence of a base such as, but not limited to, N,N-diisopropylethylamine, to provide compound (139). The reaction with tris(2-carboxyethyl))phosphine hydrochloride is typically performed at ambient temperature in a solvent such as, but not limited to, tetrahydrofuran, water, or mixtures thereof, and the reaction with N-succinimidyl 6-maleimidohexanoate is typically performed at ambient temperature in a solvent such as, but not limited to, N,N-dimethylformamide.

5.2.8. Synthesis of Compound (149)

Scheme 22 describes the synthesis of galactoside linker intermediates and synthons. (2S,3R,4S,5S,6R)-6-(Acetoxymethyl)tetrahydro-2H-pyran-2,3,4,5-tetrayl tetraacetate (140) can be treated with HBr in acetic acid to provide (2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-bromotetrahydro-2H-pyran-3,4,5-triyl triacetate (141). The reaction is typically performed at ambient temperature under a nitrogen atmosphere. (2R,3S,4S,5R,6S)-2-(Acetoxymethyl)-6-(4-formyl-2-nitrophenoxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate (143) can be prepared by treating (2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-bromotetrahydro-2H-pyran-3,4,5-triyl triacetate (141) with silver(I) oxide in the presence of 4-hydroxy-3-nitrobenzaldehyde (142). The reaction is typically performed at ambient temperature in a solvent such as, but not limited to, acetonitrile. (2R,3S,4S,5R,6S)-2-(Acetoxymethyl)-6-(4-formyl-2-nitrophenoxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate (143) can be treated with sodium borohydride to provide (2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(4-(hydroxymethyl)-2-nitrophenoxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate (144). The reaction is typically performed at low temperature in a solvent such as but not limited to tetrahydrofuran, methanol, or mixtures thereof. (2R,3S,4S,5R,6S)-2-(Acetoxymethyl)-6-(2-amino-4-(hydroxymethyl)phenoxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate (145) can be prepared by treating (2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(4-(hydroxymethyl)-2-nitrophenoxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate (144) with zinc in the presence of hydrochloric acid. The reaction is typically performed at low temperature, under a nitrogen atmosphere, in a solvent such as, but not limited to, tetrahydrofuran. (2S,3R,4S,5S,6R)-2-(2-(3-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)propanamido)-4-(hydroxymethyl)phenoxy)-6-(acetoxymethyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (146) can be prepared by reacting (2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(2-amino-4-(hydroxymethyl)phenoxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate (145) with (9H-fluoren-9-yl)methyl (3-chloro-3-oxopropyl)carbamate (103) in the presence of a base such as, but not limited to, N,N-diisopropylethylamine. The reaction is typically performed at low temperature, in a solvent such as, but not limited to, dichloromethane. (2S,3R,4S,5S,6R)-2-(2-(3-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)propanamido)-4-(hydroxymethyl)phenoxy)-6-(acetoxymethyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (146) can be reacted with bis(4-nitrophenyl)carbonate in the presence of a base such as, but not limited to, N,N-diisopropylethylamine, to provide (2S,3R,4S,5S,6R)-2-(2-(3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propanamido)-4-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenoxy)-6-(acetoxymethyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (147). The reaction is typically performed at low temperature, in a solvent such as, but not limited to, N,N-dimethylformamide. (2S,3R,4S,5S,6R)-2-(2-(3-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)propanamido)-4-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenoxy)-6-(acetoxymethyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (147) can be reacted with compound (88) in the presence of a base such as, but not limited to N,N-diisopropylethylamine, followed by treatment with lithium hydroxide, to provide compound (148). The first step is typically performed at low temperature, in a solvent such as, but not limited to, N,N-dimethylformamide, and the second step is typically performed at ambient temperature, in a solvent such as, but not limited to, methanol. Compound (148) can be treated with compound (84), wherein Sp is a spacer, in the presence of a base, such as, but not limited to N,N-diisopropylethylamine, to provide compound (149). The reaction is typically performed at ambient temperature, in a solvent such as, but not limited to, N,N-dimethylformamide.

›D-L-R x (III) · 12 of 16

III.A.7. General Methods for Synthesizing Anti-B7-H3 ADCs

The present invention also discloses a process to prepare an anti-B7-H3 ADC according to structural formula (I):

(D-L-LK m Ab (I)

wherein D, L, LK, Ab and m are as defined in the Detailed Description section. The process comprises:

treating an antibody in an aqueous solution with an effective amount of a disulfide reducing agent at 30-40° C. for at least 15 minutes, and then cooling the antibody solution to 20-27° C.;

adding to the reduced antibody solution a solution of water/dimethyl sulfoxide comprising a synthon selected from the group of 2.1 to 2.176 (Table B);

adjusting the pH of the solution to a pH of 7.5 to 8.5;

allowing the reaction to run for 48 to 80 hours to form the ADC;

wherein the mass is shifted by 18±2 amu for each hydrolysis of a succinimide to a succinamide as measured by electron spray mass spectrometry; and

wherein the ADC is optionally purified by hydrophobic interaction chromatography.

In certain embodiments, the antibody is an hB7-H3 antibody, wherein the hB7-H3 antibody comprises the heavy and light chain CDRs of huAb3v2.5, huAb3v2.6, or huAb13v1.

The present invention is also directed to an anti-B7-H3 ADC prepared by the above-described process.

In certain embodiments, the anti-B7-H3 ADC disclosed in the present application is formed by contacting an antibody that binds an hB7-H3 cell surface receptor or tumor associated antigen expressed on a tumor cell with a drug-linker synthon under conditions in which the drug-linker synthon covalently links to the antibody through a maleimide moiety as shown in formulae (IIe) and (IIf), or through an acetyl halide as shown in (IIg), or through a vinyl sulfone as shown in (IIh).

wherein D is the Bcl-xL inhibitor drug according to structural formula (IIa), (IIb), (IIc) or (IId) as described above and L 1 is the portion of the linker not formed from the maleimide, acetyl halide or vinyl sulfone upon attachment of the synthon to the antibody; and wherein the drug-linker synthon is selected from the group consisting of synthon examples 2.1 to 2.176 (Table B), or a pharmaceutically acceptable salt thereof.

In certain embodiments, the contacting step is carried out under conditions such that the anti-B7-H3 ADC has a DAR of 2, 3 or 4.

III.B. Anti-B7-H3 ADCs: Other Exemplary Drugs for Conjugation

Anti-B7-H3 antibodies may be used in ADCs to target one or more drug(s) to a cell of interest, e.g., a cancer cell expressing B7-H3. The anti-B7-H3 ADCs of the invention provide a targeted therapy that may, for example, reduce the side effects often seen with anti-cancer therapies, as the one or more drug(s) is delivered to a specific cell.

Auristatins

Anti-B7-H3 antibodies of the invention, e.g., the huAb13v1, huAb3v2.5, or huAb3v2.6 antibody, may be conjugated to at least one auristatin. Auristatins represent a group of dolastatin analogs that have generally been shown to possess anticancer activity by interfering with microtubule dynamics and GTP hydrolysis, thereby inhibiting cellular division. For example, auristatin E (U.S. Pat. No. 5,635,483) is a synthetic analogue of the marine natural product dolastatin 10, a compound that inhibits tubulin polymerization by binding to the same site on tubulin as the anticancer drug vincristine (G. R. Pettit, Prog. Chem. Org. Nat. Prod, 70: 1-79 (1997)). Dolastatin 10, auristatin PE, and auristatin E are linear peptides having four amino acids, three of which are unique to the dolastatin class of compounds. Exemplary embodiments of the auristatin subclass of mitotic inhibitors include, but are not limited to, monomethyl auristatin D (MMAD or auristatin D derivative), monomethyl auristatin E (MMAE or auristatin E derivative), monomethyl auristatin F (MMAF or auristatin F derivative), auristatin F phenylenediamine (AFP), auristatin EB (AEB), auristatin EFP (AEFP), and 5-benzoylvaleric acid-AE ester (AEVB). The synthesis and structure of auristatin derivatives are described in U.S. Patent Application Publication Nos. 2003-0083263, 2005-0238649 and 2005-0009751; International Patent Publication No. WO 04/010957, International Patent Publication No. WO 02/088172, and U.S. Pat. Nos. 6,323,315; 6,239,104; 6,034,065; 5,780,588; 5,665,860; 5,663,149; 5,635,483; 5,599,902; 5,554,725; 5,530,097; 5,521,284; 5,504,191; 5,410,024; 5,138,036; 5,076,973; 4,986,988; 4,978,744; 4,879,278; 4,816,444; and 4,486,414, each of which is incorporated by reference herein.

In one embodiment, anti-B7-H3 antibodies of the invention, e.g., huAb13v1, huAb3v2.5, or huAb3v2.6, are conjugated to at least one MMAE (mono-methyl auristatin E). Monomethyl auristatin E (MMAE, vedotin) inhibits cell division by blocking the polymerization of tubulin. However, due to its super toxicity, auristatin E cannot be used as a drug itself. Auristatin E can be linked to a monoclonal antibody (mAb) that recognizes a specific marker expression in cancer cells and directs MMAE to the cancer cells. In one embodiment, the linker linking MMAE to the anti-B7-H3 antibody is stable in extracellular fluid (i.e., the medium or environment that is external to cells), but is cleaved by cathepsin once the ADC has bound to the specific cancer cell antigen and entered the cancer cell, thus releasing the toxic MMAE and activating the potent anti-mitotic mechanism.

In one embodiment, an anti-B7-H3 antibody described herein, e.g., huAb13v1, huAb3v2.5, or huAb3v2.6, is conjugated to at least one MMAF (monomethylauristatin F). Monomethyl auristatin F (MMAF) inhibits cell division by blocking the polymerization of tubulin. It has a charged C-terminal phenylalanine residue that attenuates its cytotoxic activity compared to its uncharged counterpart MMAE. However, due to its super toxicity, auristatin F cannot be used as a drug itself, but can be linked to a monoclonal antibody (mAb) that directs it to the cancer cells. In one embodiment, the linker to the anti-B7-H3 antibody is stable in extracellular fluid, but is cleaved by cathepsin once the conjugate has entered a tumor cell, thus activating the anti-mitotic mechanism.

›D-L-R x (III) · 13 of 16

The structures of MMAF and MMAE are provided below.

An example of huAb13v1, huAb3v2.5, or huAb3v2.6-vcMMAE is also provided in FIG. 3 . Notably, FIG. 3 describes a situation where the antibody (e.g., huAb13v1, huAb3v2.5, or huAb3v2.6) is coupled to a single drug and, therefore, has a DAR of 1. In certain embodiments, the ADC will have a DAR of 2 to 8, or, alternatively, 2 to 4.

Other Drugs for Conjugation

Examples of drugs that may be used in ADCs, i.e., drugs that may be conjugated to the anti-B7-H3 antibodies of the invention, are provided below, and include mitotic inhibitors, antitumor antibiotics, immunomodulating agents, gene therapy vectors, alkylating agents, antiangiogenic agents, antimetabolites, boron-containing agents, chemoprotective agents, hormone agents, glucocorticoids, photoactive therapeutic agents, oligonucleotides, radioactive isotopes, radiosensitizers, topoisomerase inhibitors, kinase inhibitors, and combinations thereof.

1. Mitotic Inhibitors

In one aspect, anti-B7-H3 antibodies may be conjugated to one or more mitotic inhibitor(s) to form an ADC for the treatment of cancer. The term “mitotic inhibitor”, as used herein, refers to a cytotoxic and/or therapeutic agent that blocks mitosis or cell division, a biological process particularly important to cancer cells. A mitotic inhibitor disrupts microtubules such that cell division is prevented, often by effecting microtubule polymerization (e.g., inhibiting microtubule polymerization) or microtubule depolymerization (e.g., stabilizing the microtubule cytoskeleton against depolymerization). Thus, in one embodiment, an anti-B7-H3 antibody of the invention is conjugated to one or more mitotic inhibitor(s) that disrupts microtubule formation by inhibiting tubulin polymerization. In another embodiment, an anti-B7-H3 antibody of the invention is conjugated to one or more mitotic inhibitor(s) that stabilizes the microtubule cytoskeleton from depolymerization. In one embodiment, the mitotic inhibitor used in the ADCs of the invention is Ixempra (ixabepilone). Examples of mitotic inhibitors that may be used in the anti-B7-H3 ADCs of the invention are provided below. Included in the genus of mitotic inhibitors are auristatins, described above.

a. Dolastatins

The anti-B7-H3 antibodies of the invention may be conjugated to at least one dolastatin to form an ADC. Dolastatins are short peptidic compounds isolated from the Indian Ocean sea hare Dolabella auricularia (see Pettit et al., J. Am. Chem. Soc., 1976, 98, 4677). Examples of dolastatins include dolastatin 10 and dolastatin 15. Dolastatin 15, a seven-subunit depsipeptide derived from Dolabella auricularia , and is a potent antimitotic agent structurally related to the antitubulin agent dolastatin 10, a five-subunit peptide obtained from the same organism. Thus, in one embodiment, the anti-B7-H3 ADC of the invention comprises an anti-B7-H3 antibody, as described herein, and at least one dolastatin. Auristatins, described above, are synthetic derivatives of dolastatin 10.

b. Maytansinoids

The anti-B7-H3 antibodies of the invention may be conjugated to at least one maytansinoid to form an ADC. Maytansinoids are potent antitumor agents that were originally isolated from members of the higher plant families Celastraceae, Rhamnaceae, and Euphorbiaceae, as well as some species of mosses (Kupchan et al, J. Am. Chem. Soc. 94:1354-1356 [1972]; Wani et al, J. Chem. Soc. Chem. Commun. 390: [1973]; Powell et al, J. Nat. Prod. 46:660-666 [1983]; Sakai et al, J. Nat. Prod. 51:845-850 [1988]; and Suwanborirux et al, Experientia 46:117-120 [1990]). Evidence suggests that maytansinoids inhibit mitosis by inhibiting polymerization of the microtubule protein tubulin, thereby preventing formation of microtubules (see, e.g., U.S. Pat. No. 6,441,163 and Remillard et al., Science, 189, 1002-1005 (1975)). Maytansinoids have been shown to inhibit tumor cell growth in vitro using cell culture models, and in vivo using laboratory animal systems. Moreover, the cytotoxicity of maytansinoids is 1,000-fold greater than conventional chemotherapeutic agents, such as, for example, methotrexate, daunorubicin, and vincristine (see, e.g., U.S. Pat. No. 5,208,020).

Maytansinoids to include maytansine, maytansinol, C-3 esters of maytansinol, and other maytansinol analogues and derivatives (see, e.g., U.S. Pat. Nos. 5,208,020 and 6,441,163, each of which is incorporated by reference herein). C-3 esters of maytansinol can be naturally occurring or synthetically derived. Moreover, both naturally occurring and synthetic C-3 maytansinol esters can be classified as a C-3 ester with simple carboxylic acids, or a C-3 ester with derivatives of N-methyl-L-alanine, the latter being more cytotoxic than the former. Synthetic maytansinoid analogues are described in, for example, Kupchan et al., J. Med. Chem., 21, 31-37 (1978).

Suitable maytansinoids for use in ADCs of the invention can be isolated from natural sources, synthetically produced, or semi-synthetically produced. Moreover, the maytansinoid can be modified in any suitable manner, so long as sufficient cytotoxicity is preserved in the ultimate conjugate molecule. In this regard, maytansinoids lack suitable functional groups to which antibodies can be linked. A linking moiety desirably is utilized to link the maytansinoid to the antibody to form the conjugate, and is described in more detail in the linker section below. The structure of an exemplary maytansinoid, mertansine (DM1), is provided below.

Representative examples of maytansinoids include, but are not limited, to DM1 (N 2′ -deacetyl-N 2′ -(3-mercapto-1-oxopropyl)-maytansine; also referred to as mertansine, drug maytansinoid 1; ImmunoGen, Inc.; see also Chari et al. (1992) Cancer Res 52:127), DM2, DM3 (N 2′ -deacetyl-N 2′ -(4-mercapto-1-oxopentyl)-maytansine), DM4 (4-methyl-4-mercapto-1-oxopentyl)-maytansine), and maytansinol (a synthetic maytansinoid analog). Other examples of maytansinoids are described in U.S. Pat. No. 8,142,784, incorporated by reference herein.

›D-L-R x (III) · 14 of 16

Ansamitocins are a group of maytansinoid antibiotics that have been isolated from various bacterial sources. These compounds have potent antitumor activities. Representative examples include, but are not limited to ansamitocin P1, ansamitocin P2, ansamitocin P3, and ansamitocin P4.

In one embodiment of the invention, an anti-B7-H3 antibody is conjugated to at least one DM1. In one embodiment, an anti-B7-H3 antibody is conjugated to at least one DM2. In one embodiment, an anti-B7-H3 antibody is conjugated to at least one DM3. In one embodiment, an anti-B7-H3 antibody is conjugated to at least one DM4.

d. Plant Alkaloids

The anti-B7-H3 antibodies of the invention may be conjugated to at least one plant alkaloid, e.g., a taxane or vinca alkaloid. Plant alkaloids are chemotherapy treatments derived made from certain types of plants. The vinca alkaloids are made from the periwinkle plant ( catharanthus rosea ), whereas the taxanes are made from the bark of the Pacific Yew tree ( taxus ). Both the vinca alkaloids and taxanes are also known as antimicrotubule agents, and are described in more detail below.

Taxanes

Anti-B7-H3 antibodies described herein may be conjugated to at least one taxane. The term “taxane” as used herein refers to the class of antineoplastic agents having a mechanism of microtubule action and having a structure that includes the taxane ring structure and a stereospecific side chain that is required for cytostatic activity. Also included within the term “taxane” are a variety of known derivatives, including both hydrophilic derivatives, and hydrophobic derivatives. Taxane derivatives include, but not limited to, galactose and mannose derivatives described in International Patent Application No. WO 99/18113; piperazino and other derivatives described in WO 99/14209; taxane derivatives described in WO 99/09021, WO 98/22451, and U.S. Pat. No. 5,869,680; 6-thio derivatives described in WO 98/28288; sulfenamide derivatives described in U.S. Pat. No. 5,821,263; and taxol derivative described in U.S. Pat. No. 5,415,869, each of which is incorporated by reference herein. Taxane compounds have also previously been described in U.S. Pat. Nos. 5,641,803, 5,665,671, 5,380,751, 5,728,687, 5,415,869, 5,407,683, 5,399,363, 5,424,073, 5,157,049, 5,773,464, 5,821,263, 5,840,929, 4,814,470, 5,438,072, 5,403,858, 4,960,790, 5,433,364, 4,942,184, 5,362,831, 5,705,503, and 5,278,324, all of which are expressly incorporated by reference. Further examples of taxanes include, but are not limited to, docetaxel (Taxotere; Sanofi Aventis), paclitaxel (Abraxane or Taxol; Abraxis Oncology), carbazitaxel, tesetaxel, opaxio, larotaxel, taxoprexin, BMS-184476, hongdoushan A, hongdoushan B, and hongdoushan C, and nanoparticle paclitaxel (ABI-007/Abraxene; Abraxis Bioscience).

In one embodiment, the anti-B7-H3 antibody of the invention is conjugated to at least one docetaxel molecule. In one embodiment, the anti-B7-H3 antibody of the invention is conjugated to at least one paclitaxel molecule.

Vinca alkaloids

In one embodiment, the anti-B7-H3 antibody is conjugated to at least one vinca alkaloid.

Vinca alkaloids are a class of cell-cycle-specific drugs that work by inhibiting the ability of cancer cells to divide by acting upon tubulin and preventing the formation of microtubules. Examples of vinca alkaloids that may be used in the ADCs of the invention include, but are not limited to, vindesine sulfate, vincristine, vinblastine, and vinorelbine.

2. Antitumor Antibiotics

Anti-B7-H3 antibodies of the invention may be conjugated to one or more antitumor antibiotic(s) for the treatment of cancer. As used herein, the term “antitumor antibiotic” means an antineoplastic drug that blocks cell growth by interfering with DNA and is made from a microorganism. Often, antitumor antibiotics either break up DNA strands or slow down or stop DNA synthesis. Examples of antitumor antibiotics that may be included in the anti-B7-H3 ADCs of the invention include, but are not limited to, actinomycines (e.g., pyrrolo[2,1-c][1,4]benzodiazepines), anthracyclines, calicheamicins, and duocarmycins, described in more detail below.

a. Actinomycins

The anti-B7-H3 antibodies of the invention may be conjugated to at least one actinomycin. Actinomycins are a subclass of antitumor antibiotics isolated from bacteria of the genus Streptomyces. Representative examples actinomycins include, but are not limited to, actinomycin D (Cosmegen [also known as actinomycin, dactinomycin, actinomycin IV, actinomycin C1], Lundbeck, Inc.), anthramycin, chicamycin A, DC-81, mazethramycin, neothramycin A, neothramycin B, porothramycin, prothracarcin B, SG2285, sibanomicin, sibiromycin, and tomaymycin. In one embodiment, the anti-B7-H3 antibody of the invention is conjugated to at least one pyrrolobenzodiazepine (PBD). Examples of PBDs include, but are not limited to, anthramycin, chicamycin A, DC-81, mazethramycin, neothramycin A, neothramycin B, porothramycin, prothracarcin B, SG2000 (SJG-136), SG2202 (ZC-207), SG2285 (ZC-423), sibanomicin, sibiromycin and tomaymycin. Thus, in one embodiment, anti-B7-H3 antibodies of the invention are conjugated to at least one actinomycin, e.g., actinomycin D, or at least one PBD, e.g., a pyrrolobenzodiazepine (PBD) dimer.

The structures of PBDs can be found, for example, in U.S. Patent Application Pub. Nos. 2013/0028917 and 2013/0028919, and in WO 2011/130598 A1, each of which are incorporated herein by reference in their entirety. The generic structure of a PBD is provided below.

PBDs differ in the number, type and position of substituents, in both their aromatic A rings and pyrrolo C rings, and in the degree of saturation of the C ring. In the B-ring, there is generally an imine (N═C), a carbinolamine (NH—CH(OH)), or a carbinolamine methyl ether (NH—CH(OMe)) at the N10-C11 position which is the electrophilic center responsible for alkylating DNA. All of the known natural products have an (S)-configuration at the chiral C11α position which provides them with a right-handed twist when viewed from the C ring towards the A ring. The PBD examples provided herein may be conjugated to the anti-B7-H3 antibodies of the invention. Further examples of PBDs which may be conjugated to the anti-B7-H3 antibodies of the invention can be found, for example, in U.S. Patent Application Publication Nos. 2013/0028917 A1 and 2013/0028919 A1, in U.S. Pat. No. 7,741,319 B2, and in WO 2011/130598 A1 and WO 2006/111759 A1, each of which are incorporated herein by reference in their entirety.

›D-L-R x (III) · 15 of 16

A representative PBD dimer having the following formula XXX may be conjugated to the anti-B7-H3 antibodies of the invention:

wherein:

R 30 is of formula XXXI:

where A is a C 5-7 aryl group, X is a group conjugated to the Linker unit selected from the group consisting of —O—, —S—, —C(O)O—, —C(O)—, —NH(C═O)—, and —N(R N )—, wherein R is selected from the group consisting of H, C 1-4 alkyl and (C 2 H 4 O) m CH 3 , where s is 1 to 3, and either:

(i) Q 1 is a single bond, and Q 2 is selected from the group consisting of a single bond and —Z—(CH 2 ) n —, where Z is selected from the group consisting of a single bond, O, S and NH and n is from 1 to 3; or

(ii) Q is —CH═CH—, and Q is a single bond;

R 130 is a C 5-10 aryl group, optionally substituted by one or more substituents selected from the group consisting of halo, nitro, cyano, C 1-12 alkoxy, C 3-20 heterocycloalkoxy, C 5-20 aryloxy, heteroaryloxy, alkylalkoxy, arylalkoxy, alkyl aryloxy, heteroarylalkoxy, alkylheteroaryloxy, C 1-7 alkyl, C 3-7 heterocyclyl and bis-oxy-C 1-3 alkylene;

R 31 and R 33 are independently selected from the group consisting of H, R x , OH, OR x , SH, SR x , NH 2 , NHR x , NR x R xx′ , nitro, Me 3 Sn and halo;

where R and R′ are independently selected from the group consisting of optionally substituted C 1-12 alkyl, C 3-20 heterocyclyl and C 5-20 aryl groups;

R 32 is selected from the group consisting of H, R x , OH, OR x , SH, SR x , NH 2 , NHR X , NHR x R xx , nitro, Me 3 Sn and halo;

either:

(a) R 34 is H, and R 11 is OH, OR xA , where R xA is C 1-4 alkyl;

(b) R 34 and R 35 form a nitrogen-carbon double bond between the nitrogen and carbon atoms to which they are bound; or

R xxx is a C 3-12 alkylene group, which chain may be interrupted by one or more heteroatoms, selected from the group consisting of O, S, NH, and an aromatic ring;

Y x and Y x′ are is selected from the group consisting of O, S, and NH;

R 31′ , R 32′ , R 33′ are selected from the same groups as R 31 , R 32 and R 33 respectively and R 34′ and R 35′ are the same as R 34 and R 35 , and each M is a monovalent pharmaceutically acceptable cation or both M groups together are a divalent pharmaceutically acceptable cation.

C 1-12 alkyl: The term “C 1-12 alkyl” as used herein, pertains to a monovalent moiety obtained by removing a hydrogen atom from a carbon atom of a hydrocarbon compound having from 1 to 12 carbon atoms, which may be aliphatic or alicyclic, and which may be saturated or unsaturated (e.g. partially unsaturated, fully unsaturated). Thus, the term “alkyl” includes the sub-classes alkenyl, alkynyl, cycloalkyl, etc., discussed below.

Examples of saturated alkyl groups include, but are not limited to, methyl (C 1 ), ethyl (C 2 ), propyl (C 3 ), butyl (C 4 ), pentyl (C 5 ), hexyl (C 6 ) and heptyl (C 7 ).

Examples of saturated linear alkyl groups include, but are not limited to, methyl (C 1 ), ethyl (C 2 ), n-propyl (C 3 ), n-butyl (C 4 ), n-pentyl (amyl) (C 5 ), n-hexyl (C 6 ) and n-heptyl (C 7 ).

Examples of saturated branched alkyl groups include iso-propyl (C 3 ), iso-butyl (C 4 ), sec-butyl (C 4 ), tert-butyl (C 4 ), iso-pentyl (C 5 ), and neo-pentyl (C 5 ).

C 3-20 heterocyclyl: The term “C 3-20 heterocyclyl” as used herein, pertains to a monovalent moiety obtained by removing a hydrogen atom from a ring atom of a heterocyclic compound, which moiety has from 3 to 20 ring atoms, of which from 1 to 10 are ring heteroatoms. Preferably, each ring has from 3 to 7 ring atoms, of which from 1 to 4 are ring heteroatoms.

In this context, the prefixes (e.g. C 3-20 , C 3-7 , C 5-6 , etc.) denote the number of ring atoms, or range of number of ring atoms, whether carbon atoms or heteroatoms. For example, the term “C 5-6 heterocyclyl”, as used herein, pertains to a heterocyclyl group having 5 or 6 ring atoms.

Examples of monocyclic heterocyclyl groups include, but are not limited to, those derived from:

N 1 : aziridine (C 3 ), azetidine (C 4 ), pyrrolidine (tetrahydropyrrole) (C 5 ), pyrroline (e.g., 3-pyrroline, 2,5-dihydropyrrole) (C 5 ), 2H-pyrrole or 3H-pyrrole (isopyrrole, isoazole) (C 5 ), piperidine (C 6 ), dihydropyridine (C 6 ), tetrahydropyridine (C 6 ), azepine (C 7 ); O 1 : oxirane (C 3 ), oxetane (C 4 ), oxolane (tetrahydrofuran) (C 5 ), oxole (dihydrofuran) (C 5 ), oxane (tetrahydropyran) (C 6 ), dihydropyran (C 6 ), pyran (C 6 ), oxepin (C 7 ); S 1 : thiirane (C 3 ), thietane (C 4 ), thiolane (tetrahydrothiophene) (C 5 ), thiane (tetrahydrothiopyran) (C 6 ), thiepane (C 7 ); O 2 : dioxolane (C 5 ), dioxane (C 6 ), and dioxepane (C 7 ); O 3 : trioxane (C 6 ); N 2 : imidazolidine (C 5 ), pyrazolidine (diazolidine) (C 5 ), imidazoline (C 5 ), pyrazoline (dihydropyrazole) (C 5 ), piperazine (C 6 ); N 1 O 1 : tetrahydrooxazole (C 5 ), dihydrooxazole (C 5 ), tetrahydroisoxazole (C 5 ), dihydroisoxazole (C 5 ), morpholine (C 6 ), tetrahydrooxazine (C 6 ), dihydrooxazine (C 6 ), oxazine (C 6 ); N 1 S 1 : thiazoline (C 5 ), thiazolidine (C 5 ), thiomorpholine (C 6 ); N 2 O 1 : oxadiazine (C 6 ); O 1 S 1 : oxathiole (C 5 ) and oxathiane (thioxane) (C 6 ); and, N 1 O 1 S 1 : oxathiazine (C 6 ).

Examples of substituted monocyclic heterocyclyl groups include those derived from saccharides, in cyclic form, for example, furanoses (C 5 ), such as arabinofuranose, lyxofuranose, ribofuranose, and xylofuranse, and pyranoses (C 6 ), such as allopyranose, altropyranose, glucopyranose, mannopyranose, gulopyranose, idopyranose, galactopyranose, and talopyranose.

C 5-20 aryl: The term “C 5-20 aryl”, as used herein, pertains to a monovalent moiety obtained by removing a hydrogen atom from an aromatic ring atom of an aromatic compound, which moiety has from 3 to 20 ring atoms. Preferably, each ring has from 5 to 7 ring atoms.

In this context, the prefixes (e.g. C 3-20 , C 5-7 , C 5-6 , etc.) denote the number of ring atoms, or range of number of ring atoms, whether carbon atoms or heteroatoms. For example, the term “C 5-6 aryl” as used herein, pertains to an aryl group having 5 or 6 ring atoms.

›D-L-R x (III) · 16 of 16

In one embodiment, the anti-B7-H3 antibodies of the invention may be conjugated to a PBD dimer having the following formula XXXIa:

wherein the above structure describes the PBD dimer SG2202 (ZC-207) and is conjugated to the anti-B7-H3 antibody of the invention via a linker L. SG2202 (ZC-207) is disclosed in, for example, U.S. Patent App. Pub. No. 2007/0173497, which is incorporated herein by reference in its entirety.

In another embodiment, a PBD dimer, SGD-1882, is conjugated to anti-B7-H3 antibody of the invention via a drug linker, as depicted in FIG. 4 . SGD-1882 is disclosed in Sutherland et al. (2013) Blood 122(8): 1455 and in U.S Patent App. Pub. No. 2013/0028919, which is incorporated herein by reference in its entirety. As described in FIG. 4 , the PBD dimer SGD-1882 may be conjugated to an antibody via an mc-val-ala-dipeptide linker (collectively referred to as SGD-1910 in FIG. 4 ). In a certain embodiment, an anti-B7-H3 antibody, as disclosed herein, is conjugated to the PBD dimer described in FIG. 4 . Thus, in a further embodiment, the invention includes an anti-B7-H3 antibody, as disclosed herein, conjugated to a PBD dimer via a mc-val-ala-dipeptide linker, as described in FIG. 4 . In certain embodiments, the invention includes an anti-B7-H3 antibody comprising a heavy chain variable region comprising a CDR3 domain comprising the amino acid sequence of SEQ ID NO: 35, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 34, and a CDR1 domain comprising the amino acid sequence

›Tables in the description — 28

TABLE A Sequences of human IgG heavy chain constant domains and light chain constant domains Sequence

Protein	Identifier	Sequence
Ig gamma-1	SEQ ID	ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDY
constant	NO: 159	FPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
region		LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK
		KVEPKSCDKTHTCPPCPAPELLGGPSVFLFPP
		KPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW
		YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL
		HQDWLNGKEYKCKVSNKALPAPIEKTISKAKG
		QPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
		PSDIAVEWESNGQPENNYKTTPPVLDSDGSFF
		LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT
		QKSLSLSPGK
Ig gamma-1	SEQ ID	ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDY
constant	NO: 160	FPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
region		LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK
mutant		KVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPP
		KPKDTLMISRTPEVICVVVDVSHEDPEVKFNW
		YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL
		HQDWLNGKEYKCKVSNKALPAPIEKTISKAKG
		QPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
		PSDIAVEWESNGQPENNYKTTPPVLDSDGSFF
		LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT
		QKSLSLSPGK
Ig Kappa	SEQ ID	RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNF
constant	NO: 161	YPREAKVQWKVDNALQSGNSQESVTEQDSKDS
region		TYSLSSTLTLSKADYEKHKVYACEVTHQGLSS
		PVTKSFNRGEC
Ig Lambda	SEQ ID	QPKAAPSVTLFPPSSEELQANKATLVCLISDF
constant	NO: 162	YPGAVTVAWKADSSPVKAGVETTTPSKQSNNK
region		YAASSYLSLTPEQWKSHRSYSCQVTHEGSTVE
		KTVAPTECS

App Ex. No.	Bcl-xL Inhibitor Cmpd No
1.1	W2.01
1.2	W2.02
1.3	W2.03
1.5	W2.05
1.6	W2.06
1.7	W2.07
1.8	W2.08
1.9	W2.09
1.10	W2.10
1.11	W2.11
1.12	W2.12
1.13	W2.13
1.14	W2.14
1.15	W2.15
1.16	W2.16
1.17	W2.17
1.18	W2.18
1.19	W2.19
1.20	W2.20
1.21	W2.21
1.22	W2.22
1.23	W2.23
1.24	W2.24
1.25	W2.25
1.26	W2.26
1.27	W2.27
1.28	W2.28
1.29	W2.29
1.30	W2.30
1.31	W2.31
1.32	W2.32
1.33	W2.33
1.34	W2.34
1.35	W2.35
1.36	W2.36
1.37	W2.37
1.38	W2.38
1.39	W2.39
1.40	W2.40
1.41	W2.41
1.42	W2.42
1.43	W2.43
1.44	W2.44
1.45	W2.45
1.46	W2.46
1.47	W2.47
1.48	W2.48
1.49	W2.49
1.50	W2.50
1.51	W2.51
1.52	W2.52
1.53	W2.53
1.54	W2.54
1.55	W2.55
1.56	W2.56
1.57	W2.57
1.58	W2.58
1.59	W2.59
1.60	W2.60
1.61	W2.61
1.62	W2.62
1.63	W2.63
1.64	W2.64
1.65	W2.65
1.66	W2.66
1.67	W2.67
1.68	W2.68
1.69	W2.69
1.70	W2.70
1.71	W2.71
1.72	W2.72
1.73	W2.73
1.74	W2.74
1.75	W2.75
1.76	W2.76
1.77	W2.77
1.78	W2.78
1.79	W2.79
1.80	W2.80
1.81	W2.81
1.82	W2.82
1.83	W2.83
1.84	W2.84
1.85	W2.85
1.86	W2.86
1.87	W2.87
1.88	W2.88
1.89	W2.89
1.90	W2.90
1.91	W2.91

TABLE B

Example	Synthon
No.	Code	Synthon Structure
2.1	CZ
2.2	DH
2.4	EP
2.5	EF
2.6	EG
2.7	EH
2.8	ER
2.9	ES
2.10	EQ
2.11	EU
2.12	EV
2.13	EW
2.14	EX
2.15	EY
2.16	EZ
2.17	FD
2.18	FS
2.19	FI
2.20	FV
2.21	GC
2.22	GB
2.23	FW
2.24	GD
2.25	GK
2.26	GJ
2.27	GW
2.28	HF
2.29	HG
2.30	HP
2.31	HR
2.32	HU
2.33	HT
2.34	HV
2.35	HZ
2.36	IA
2.37	IF
2.38	IG
2.39	IH
2.40	IJ
2.41	IK
2.42	IL
2.43	IM
2.44	IO
2.45	IP
2.46	IS
2.47	IU
2.48	IV
2.49	IZ
2.50	JD
2.51	JF
2.52	JK
2.53	JJ
2.54	JL
2.55	FE
2.56	GG
2.57	GM
2.58	HD
2.59	HS
2.60	HW
2.61	HX
2.62	HY
2.63	IB
2.64	IE
2.65	II
2.66	KY
2.67	IW
2.68	IY
2.69	JA
2.77	FA
2.78	FJ
2.79	FK
2.80	FQ
2.81	FR
2.82	JE
2.83	JM
2.84	LE
2.85	LH
2.86	LJ
2.87	MA
2.88	MD
2.89	MG
2.90	MS
2.91	MR
2.92	MQ
2.93	MZ
2.94	NA
2.95	NB
2.96	NP
2.97	NN
2.98	NO
2.101	OK
2.102	OW
2.103	PC
2.104	PI
2.105	PJ
2.106	PU
2.107	PV
2.108	PW
2.109	QW
2.110	RM
2.111	RR
2.112	SJ
2.113	SM
2.114	SN
2.115	SS
2.116	TA
2.117	TW
2.118	ST
2.119	ZL
2.120	SX
2.121	SW
2.122	TV
2.123	SZ
2.124	ZM
2.125	SV
2.126	SY
2.127	TK
2.128	TR
2.129	TY
2.130	TX
2.131	TZ
2.132	UA
2.133	UJ
2.134	UK
2.135	UU
2.136	UV
2.137	UZ
2.138	VB
2.139	VC
2.140	VS
2.141	VT
2.142	VY
2.143	WI
2.144	WK
2.145	WP
2.146	XD
2.147	XK
2.148	XL
2.149	YJ
2.150	YQ
2.151	YR
2.152	YS
2.153	YY
2.154	YT
2.155	YU
2.156	YV
2.157	YW
2.158	ZB
2.159	ZC
2.160	ZJ
2.161	ZE
2.162	ZS
2.163	ZW
2.164	ZX
2.166	AAA
2.167	AAD
2.168	AAE
2.169	ABG
2.170	ABL
2.171	ABN
2.172	AAF
2.173	ABO
2.174	ABM
2.175	ABU
2.176	ABV
2.177 (control)	LB
2.178 (control)	WD
2.179 (control)	ZZ
2.180 (control)	ZT
2.181 (control)	XW
2.182 (control)	SE
2.183 (control)	SR
2.184 (control)	YG
2.185 (control)	KZ

Anti-Cancer Agent	Comments	Examples
Antibodies	Antibodies which bind IGF-	A12 (fully humanized mAb)
	1R (insulin-like growth	19D12 (fully humanized mAb)
	factor type 1 receptor),	Cp751-871 (fully humanized mAb)
	which is expressed on the	H7C10 (humanized mAb)
	cell surface of most human	alphaIR3 (mouse)
	cancers	ScFV/FC (mouse/human chimera)
		EM/164 (mouse)
	Antibodies which bind	Matuzumab (EMD72000)
	EGFR; Mutations affecting	Erbitux ®/Cetuximab (Imclone)
	EGFR expression or activity	Vectibix ®/Panitumumab (Amgen)
	could result in cancer	mAb 806
		Nimotuxumab (TheraCIM)
	Antibodies which bind	AVEO (AV299) (AVEO)
	cMET (Mesechymal	AMG102 (Amgen)
	epithelial transition factor);	5D5 (OA-5d5) (Genentech)
	a member of the MET	H244G11 (Pierre Fabre)
	family of receptor tyrosine
	kinases)
	Anti-ErbB3 antibodies	Ab #14 (MM 121-14)
		Herceptin ® (Trastuzumab; Genentech)
		1B4C3; 2D1D12 (U3 Pharma AG)
Small Molecules	Insulin-like growth factor	NVP-AEW541-A
Targeting IGF1R	type 1 receptor which is	BMS-536,924 (1H-benzoimidazol-2-yl)-1H-
	expressed on the cell	pyridin-2-one)
	surface of many human	BMS-554,417
	cancers	Cycloligan
		TAE226
		PQ401
Small Molecules	EGFR;	Iressa ®/Gefitinib (AstraZeneca)
Targeting EGFR	Overexpression or	CI-1033 (PD 183805) (Pfizer)
	mutations affecting EGFR	Lapatinib (GW-572016) (GlaxoSmithKline)
	expression or activity could	Tykerb ®/Lapatinib Ditosylate (Smith Kline
	result in cancer	Beecham)
		Tarceva ®/Erlotinib HCL (OSI-774) (OSI
		Pharma)
		PKI-166 (Novartis)
		PD-158780
		EKB-569
		Tyrphostin AG 1478 (4-(3-Chloroanillino)-
		6,7-dimethoxyquinazoline)
Small Molecules	cMET (Mesenchymal	PHA665752
Targeting cMET	epithelial transition factor);	ARQ 197
	a member of the MET
	family of receptor tyrosine
	kinases)
Antimetabolites		Flourouracil (5-FU)
		Capecitabine/XELODA ® (HLR Roche)
		5-Trifluoromethyl-2′-deoxyuridine
		Methotrexate sodium (Trexall) (Barr)
		Raltitrexed/Tomudex ® (AstraZeneca)
		Pemetrexed/Alimta ® (Lilly)
		Tegafur
		Cytosine Arabinoside (Cytarabine, Ara-C)/
		Thioguanine ® (GlaxoSmithKline)
		5-azacytidine
		6-mercaptopurine (Mercaptopurine, 6-MP)
		Azathioprine/Azasan ® (AAIPHARMA LLC)
		6-thioguanine (6-TG)/Purinethol ® (TEVA)
		Pentostatin/Nipent ® (Hospira Inc.)
		Fludarabine phosphate/Fludara ® (Bayer
		Health Care)
		Cladribine (2-CdA, 2-chlorodeoxyadenosine)/
		Leustatin ® (Ortho Biotech)
Alkylating agents	An alkylating antineoplastic	Ribonucleotide Reductase Inhibitor (RNR)
	agent is an alkylating agent	Cyclophosphamide/Cytoxan (BMS)
	that attaches an alkyl group	Neosar (TEVA)
	to DNA. Since cancer cells	Ifosfamide/Mitoxana ® (ASTA Medica)
	generally proliferate	Thiotepa (Bedford, Abraxis, Teva)
	unrestrictively more than do	BCNU→ 1,3-bis(2-chloroethyl)-1-nitosourea
	healthy cells they are more	CCNU→ 1, -(2-chloroethyl)-3-cyclohexyl-1-
	sensitive to DNA damage,	nitrosourea (methyl CCNU)
	and alkylating agents are	Hexamethylmelamine (Altretamine, HMM)/
	used clinically to treat a	Hexalen ® (MGI Pharma Inc.)
	variety of tumors.	Busulfan/Myleran (GlaxoSmithKline)
		Procarbazine HCL/Matulane (Sigma Tau
		Pharmaceuticals, Inc.)
		Dacarbazine (DTIC)
		Chlorambucil/Leukara ® (SmithKline
		Beecham)
		Melphalan/Alkeran ® (GlaxoSmithKline)
		Cisplatin (Cisplatinum, CDDP)/Platinol
		(Bristol Myers)
		Carboplatin/Paraplatin (BMS)
		Oxaliplatin/Eloxitan ® (Sanofi-Aventis US)
Topoisomerase	Topoisomerase inhibitors	Doxorubicin HCL/Doxil ® (Alza)
inhibitors	are chemotherapy agents	Daunorubicin citrate/Daunoxome ® (Gilead)
	designed to interfere with	Mitoxantrone HCL/Novantrone (EMD
	the action of topoisomerase	Serono)
	enzymes (topoisomerase I	Actinomycin D
	and II), which are enzymes	Etoposide/Vepesid ® (BMS)/Etopophos ®
	that control the changes in	(Hospira, Bedford, Teva Parenteral, Etc.)
	DNA structure by	Topotecan HCL/Hycamtin ®
	catalyzing the breaking and	(GlaxoSmithKline)
	rejoining of the	Teniposide (VM-26)/Vumon ® (BMS)
	phosphodiester backbone of	Irinotecan HCL(CPT-ll/Camptosar ®
	DNA strands during the	(Pharmacia & Upjohn)
	normal cell cycle.
Microtubule	Microtubules are one of the	Vincristine/Oncovin ® (Lilly)
targeting agents	components of the	Vinblastine sulfate/Velban ®(discontinued)
	cytoskeleton. They have	(Lilly)
	diameter of ~24 nm and	Vinorelbine tartrate/Navelbine ®
	length varying from several	(PierreFabre)
	micrometers to possibly	Vindesine sulphate/Eldisine ® (Lilly)
	millimeters in axons of	Paclitaxel/Taxol ® (BMS)
	nerve cells. Microtubules	Docetaxel/Taxotere ® (Sanofi Aventis US)
	serve as structural	Nanoparticle paclitaxel (ABI-007)/
	components Within cells and	Abraxane ® (Abraxis BioScience, Inc.)
	are involved in many	Ixabepilone/IXEMPRA ™ (BMS)
	cellular processes including
	mitosis, cytokinesis, and
	vesicular transport.
Kinase inhibitors	Kinases are enzymes that	Imatinib mesylate/Gleevec (Novartis)
	catalyze the transfer of	Sunitinib malate/Sutent ® (Pfizer)
	phosphate groups from	Sorafenib tosylate/Nexavar ® (Bayer)
	high-energy, phosphate-	Nilotinib hydrochloride monohydrate/
	donating molecules to	Tasigna ® (Novartis), Osimertinib,
	specific substrates, and are	Cobimetinib, Trametinib, Dabrafenib,
	utilized to transmit signals	Dinaciclib
	and regulate complex
	processes in cells.
Protein synthesis	Induces cell apoptosis	L-asparaginase/Elspar ® (Merck & Co.)
inhibitors
Immunotherapeutic	Induces cancer patients to	Alpha interferon
agents	exhibit immune	Angiogenesis Inhibitor/Avastin ®
	responsiveness	(Genentech)
		IL-2→ Interleukin 2 (Aldesleukin)/Proleukin ®
		(Chiron)
		IL-12→ Interleukin 12
	Antibody/small molecule	Anti-CTLA-4 and PR-1 therapies
	immune checkpoint	Yervoy ® (ipilimumab; Bristol-Myers Squibb)
	modulators	Opdivo ® (nivolumab; Bristol-Myers Squibb)
		Keytrada ® (pembrolizumab; Merck)
Hormones	Hormone therapies	Toremifene citrate/Fareston ® (GTX, Inc.)
	associated with menopause	Fulvestrant/Faslodex ® (AstraZeneca)
	and aging seek to increase	Raloxifene HCL/Evista ® (Lilly)
	the amount of certain	Anastrazole/Arimidex ® (AstraZeneca)
	hormones in your body to	Letrozole/Femara ® (Novartis)
	compensate for age- or	Fadrozole (CGS 16949A)
	disease-related hormonal	Exemestane/Aromasin ® (Pharmacia &
	declines. Hormone therapy	Upjohn)
	as a cancer treatment either	Leuprolide acetate/Eligard ® (QTL USA)
	reduces the level of specific	Lupron ® (TAP Pharm)
	hormones or alters the	Goserelin acetate/Zoladex ® (AstraZeneca)
	cancer's ability to use these	Triptorelin pamoate/Trelstar ® (Watson Labs)
	hormones to grow and	Buserelin/Suprefact ® (Sanofi Aventis)
	spread.	Nafarelin/Synarel ® (Pfizer)
		Cetrorelix/Cetrotide ® (EMD Serono)
		Bicalutamide/Casodex ® (AstraZeneca)
		Nilutamide/Nilandron ® (Aventis Pharm.)
		Megestrol acetate/Megace ® (BMS)
		Somatostatin Analogs (Octreotide acetate/
		Sandostatin ® (Novartis)
Glucocorticoids	Anti-inflammatory drugs	Prednisolone
	used to reduce swelling that	Dexamethasone/Decadron ® (Wyeth)
	causes cancer pain.
Aromatose inhibitors	Includes imidazoles	Ketoconazole
mTOR inhibitors	the mTOR signaling	Sirolimus (Rapamycin)/Rapamune ® (Wyeth)
	pathway was originally	Temsirolimus (CCI-779)/Torisel ® (Wyeth)
	discovered during studies of	Deforolimus (AP23573)/(Ariad Pharm.)
	the immunosuppressive	Everolimus (RAD00I)/Certican ® (Novartis)
	agent rapamycin. This
	highly conserved pathway
	regulates cell proliferation
	and metabolism in response
	to environmental factors,
	linking cell growth factor
	receptor signaling via
	phosphoinositide-3-
	kinase(PI-3K) to cell
	growth, proliferation, and
	angiogenesis.

TABLE 2 — List of Anti-B7-H3 antibodies generated using mouse hybridoma technology FACS Binding (EC 50 nM)

Clone			Cynomolgous
Name	Species/Isotype	Human B7-H3	B7-H3	Mouse B7-H3
Ab1	mouse IgG1/k	2.10	1.79	299.0
Ab2	mouse IgG1/k	1.70	1.50	1.00
Ab3	mouse IgG1/k	1.66	1.42	0.94
Ab4	mouse IgG2b/k	4.06	3.10	1.75
Ab5	mouse IgG1/k	2.71	1.91	6.01
Ab6	mouse IgG1/k	1.59	1.53	No binding
Ab7	mouse IgG1/k	3.22	2.67	67.13
Ab8	mouse IgG1/k	3.83	8.63	193.0
Ab9	mouse IgG1/k	4.49	259.0	0.72
Ab10	mouse IgG2b/k	3.97	4.46	3.80
Ab11	mouse IgG1/k	23.40	2.03	568.60
Ab12	mouse IgG1/k	3.88	6.71	8.72
Ab13	mouse IgG1/k	1.94	4.12	25.80
Ab14	mouse IgG1/k	3.03	2.97	102.2
Ab15	mouse IgG1/k	5.37	6.52	4.61
Ab16	mouse IgG1/k	3.94	4.28	318.7
Ab17	mouse IgG2b/k	2.75	2.60	2.39
Ab18	mouse IgG1/k	5.98	6.49	No binding

TABLE 3 — Biacore kinetics of anti-B7-H3 mouse hybridoma antibodies binding to human and cynomolgus monkey B7-H3. Murine

Antibody	huB7-H3	cynoB7-H3
Name	k a (1/Ms)	k d (1/s)	K D (M)	k a (1/Ms)	k d (1/s)	K D (M)
Ab17	5.4E+05	1.9E−05	3.4E−11	5.1E+05	1.0E−05	1.9E−11
Ab18	2.1E+05	3.6E−05	1.7E−10	2.4E+05	2.9E−05	1.2E−10
Ab15	8.0E+04	3.4E−05	4.3E−10	7.7E+04	7.0E−05	9.1E−10
Ab4	6.9E+05	1.1E−03	1.6E−09	5.4E+05	9.6E−04	1.8E−09
Ab8	5.8E+04	9.9E−05	1.7E−09	1.6E+05	2.6E−04	1.7E−09
Ab10	4.1E+04	1.9E−04	4.6E−09	2.0E+05	4.2E−03	2.0E−08
Ab12	3.8E+04	2.5E−04	6.7E−09	5.5E+04	1.0E−05	1.8E−10
Ab5	1.3E+06	1.2E−02	9.2E−09	1.4E+06	2.8E−01	2.0E−07
Ab14	1.1E+05	1.4E−03	1.3E−08	6.9E+05	3.0E−03	4.3E−09
Ab9	6.6E+04	1.1E−03	1.7E−08	poor kinetic fit
Ab13	3.3E+05	5.8E−03	1.7E−08	4.4E+05	3.7E−03	8.4E−09
Ab3	5.2E+05	1.0E−02	1.9E−08	3.8E+05	1.0E−02	2.6E−08
Ab16	1.4E+05	3.2E−03	2.4E−08	7.5E+05	5.6E−03	7.5E−09
Ab2	1.2E+05	2.9E−03	2.4E−08	2.3E+05	1.1E−02	5.0E−08
Ab11	2.0E+04	8.9E−04	4.5E−08	2.7E+04	7.2E−05	2.6E−09
Ab6	1.2E+04	1.0E−02	8.4E−07	2.8E+04	1.2E−02	4.1E−07
Ab1	no	no
	observable	observable
	binding	binding
Ab7	little	little
	observable	observable
	binding	binding

TABLE 4 — Variable region amino acid sequences of anti-B7-H3 antibodies from mouse hybridomas

SEQ		Protein
ID NO:	Clone	Region	Residues	Amino Acid Sequence
1	chAb2	VH		QVQLQQPGAELVKPGASVKLSCKAS GY
				TFTSYWMH WVKQRPGQGLEWIG MIHPD
				SGTTNYNEKFRS KATLTVDKSSSTAYM
				QLSSLTSEDSAVYYCAV YYGSTYWYFD
				V WGTGTTVTVSS
2	chAb2	CDR-H1	Residues 26-35
GYTFTSYWMH
			of SEQ ID NO: 1
3	chAb2	CDR-H2	Residues 50-66
MIHPDSGTTNYNEKFRS
			of SEQ ID NO: 1
4	chAb2	CDR-H3	Residues 99-109
YYGSTYWYFDV
			of SEQ ID NO: 1
5	chAb2	VL		DVVMTQTPLSLPVSLGDQAYISC RSSQ
				SLVHINGNTYLH WYRQKPGQSPKLLIY
				KVSNRFS GVPDRFSGSGSGTDFTLKIS
				RVEAEDLGVYFC SQSTHFPFT FGSGTK
				LEIK
6	chAb2	CDR-L1	Residues 24-39
RSSQSLVHINGNTYLH
			of SEQ ID NO: 5
7	chAb2	CDR-L2	Residues 55-61
KVSNRFS
			of SEQ ID NO: 5
8	chAb2	CDR-L3	Residues 94-102
SQSTHFPFT
			of SEQ ID NO: 5
9	chAb3	VH		QVQLQQPGAELVKPGASVKLSCKAS GY
				TFSSYWMH WVKQRPGQGLEWIG LIHPD
				SGSTNYNEMFKN KATLTVDRSSSTAYV
				QLSSLTSEDSAVYFCAG GGRLYFDY WG
				QGTTLTVSS
10	chAb3	CDR-H1	Residues 26-35
GYTFSSYWMH
			of SEQ ID NO: 9
11	chAb3	CDR-H2	Residues 50-66
LIHPDSGSTNYNEMFKN
			of SEQ ID NO: 9
12	chAb3	CDR-H3	Residues 99-106
GGRLYFDY
			of SEQ ID NO: 9
13	chAb3	VL		DVVMTQTPLSLPVSLGDQASISC RSSQ
				SLVHSNGDTYLR WYLQKPGQSPKLLIY
				KVSNRFS GVPDRFSGSGSGTDFTLKIT
				RVEAEDLGVYFC SQSTHVPYT FGGGTK
				LEIK
14	chAb3	CDR-L1	Residues 24-39
RSSQSLVHSNGDTYLR
			of SEQ ID NO: 13
7	chAb3	CDR-L2	Residues 55-61
KVSNRFS
			of SEQ ID NO: 13
15	chAb3	CDR-L3	Residues 94-102
SQSTHVPYT
			of SEQ ID NO: 13
16	chAb4	VH		QVQLQQPGAELVKPGASVKLSCKAS GY
				SFTSYWMH WVKQRPGQGLEWIG MIHPN
				SGSNNYNEKFKS KATLTVDKSSNTAYM
				QLSSLTSEDSAVYYCAR RLGLHFDY WG
				QGTTLTVSS
17	chAb4	CDR-H1	Residues 26-35
GYSFTSYWMH
			of SEQ ID NO: 16
18	chAb4	CDR-H2	Residues 50-66
MIHPNSGSNNYNEKFKS
			of SEQ ID NO: 16
19	chAb4	CDR-H3	Residues 99-106
RLGLHFDY
			of SEQ ID NO: 16
20	chAb4	VL		DIVMTQSQKFMSTPVGDRVSITC KASQ
				NVGTAVA WYQQKPGQSPKLLIY SASNR
				YT GVPDRFTGSGSGTDFTLTISNMQSE
				DLADYFC QQYSSYPYT FGGGTKLEIK
21	chAb4	CDR-L1	Residues 24-34
KASQNVGTAVA
			of SEQ ID NO: 20
22	chAb4	CDR-L2	Residues 50-56
SASNRYT
			of SEQ ID NO: 20
23	chAb4	CDR-L3	Residues 89-97
QQYSSYPYT
			of SEQ ID NO: 20
24	chAb18	VH		QVQLQQSAAELARPGASVKMSCKAS GY
				SFTSYTIH WVKQRPGQGLEWIG YINPN
				SRNTDYNQKFKD ETTLTADRSSSTAYM
				QLISLTSEDSAVYYCAR YSGSTPYWYF
				DV WGAGTTVTVSS
25	chAb18	CDR-H1	Residues 26-35
GYSFTSYTIH
			of SEQ ID NO: 24
26	chAb18	CDR-H2	Residues 50-66
YINPNSRNTDYNQKFKD
			of SEQ ID NO: 24
27	chAb18	CDR-H3	Residues 99-110
YSGSTPYWYFDV
			of SEQ ID NO: 24
28	chAb18	VL		QIVLTQSPAILSASPGEKVTMTC RASS
				SVSYMN WYQQKPGSSPKPWIY ATSNLA
				S GVPARFSVSVSGTSHSLTISRVEAED
				AATYYC QQWSSNPLT FGAGTKLELK
29	chAb18	CDR-L1	Residues 24-33
RASSSVSYMN
			of SEQ ID NO: 28
30	chAb18	CDR-L2	Residues 49-55
ATSNLAS
			of SEQ ID NO: 28
31	chAb18	CDR-L3	Residues 88-96
QQWSSNPLT
			of SEQ ID NO: 28
32	chAb13	VH		DVQLQESGPDLVKPSQSLSLTCTVT GY
				SITSGYSWH WIRQFPGNKLEWMG YIHS
				SGSTNYNPSLKS RISINRDTSKNQFFL
				QLNSVTTEDTATYYCAG YDDYFEY WGQ
				GTTLTVSS
33	chAb13	CDR-H1	Residues 26-36
GYSITSGYSWH
			of SEQ ID NO: 32
34	chAb13	CDR-H2	Residues 51-66
YIHSSGSTNYNPSLKS
			of SEQ ID NO: 32
35	chAb13	CDR-H3	Residues 99-105
YDDYFEY
			of SEQ ID NO: 32
36	chAb13	VL		DIVMTQSQKFMSTSVGDRVSVTC KASQ
				NVGFNVA WYQQKPGQSPKALIY SASYR
				YS GVPDRFTGSGSGTDFTLTISNVQSE
				DLAEYFC QQYNSYPFT FGSGTKLEIK
37	chAb13	CDR-L1	Residues 24-34
KASQNVGFNVA
			of SEQ ID NO: 36
38	chAb13	CDR-L2	Residues 50-56
SASYRYS
			of SEQ ID NO: 36
182	enAb13	CDR-L3	Residues 89-97
QQYNSYPFT
			of SEQ ID NO: 36
40	chAb12	VH		EVQLVESGGGLVKPGGSLKLSCAAS GF
				TFSSYAMS WVRQTPEKRLEWVA TISSG
				TNYTYYPDSVKG RFTISRDNAKNTLYL
				QMTSLRSEDTAMYYCAR QGRYSWIAY W
				GQGTLVTVSA
41	chAb12	CDR-H1	Residues 26-35
GFTFSSYAMS
			of SEQ ID NO: 40
42	chAb12	CDR-H2	Residues 50-66
TISSGTNYTYYPDSVKG
			of SEQ ID NO: 40
43	chAb12	CDR-H3	Residues 99-107
QGRYSWIAY
			of SEQ ID NO: 40
44	chAb12	VL		DIVLTQSPASLAVSLGQRATISC RASK
				SVSTSDYSYMH WNQQKPGQPPKLLIY L
				ASNLES GVPARFSGSGSGTDFTLNIHP
				VEEEDAATYYC QHSRELLT FGAGTKLE
				LK
45	chAb12	CDR-L1	Residues 24-38
RASKSVSTSDYSYMH
			of SEQ ID NO: 44
46	chAb12	CDR-L2	Residues 54-60
LASNLES
			of SEQ ID NO: 44
47	chAb12	CDR-L3	Residues 93-100
QHSRELLT
			of SEQ ID NO: 44
48	chAb14	VH		EVKLVESGGGLVKPGGSLKLSCAAS GF
				TFSSYGMS WVRQTPEKRLEWVA TISGG
				GTNTYYPDSVEG RFTISRDNAKNFLYL
				QMSSLRSEDTALYYCAR HYGSQTMDY W
				GQGTSVTVSS
49	chAb14	CDR-H1	Residues 26-35
GFTFSSYGMS
			of SEQ ID NO: 48
50	chAb14	CDR-H2	Residues 50-66
TISGGGTNTYYPDSVEG
			of SEQ ID NO: 48
51	chAb14	CDR-H3	Residues 99-107
HYGSQTMDY
			of SEQ ID NO: 48
52	chAb14	VL		DIQMTQSPASLSASVGETVTITC RTSG
				NIHNYLT WYQQKQGKSPQLLVY NAKTL
				AD GVPSRFSGSGSGTQFSLKINSLQPE
				DFGSYYC QHFWSIMWT FGGGTKLEIK
53	chAb14	CDR-L1	Residues 24-34
RTSGNIHNYLT
			of SEQ ID NO: 52
54	chAb14	CDR-L2	Residues 50-56
NAKTLAD
			of SEQ ID NO: 52
55	chAb14	CDR-L3	Residues 89-97
QHFWSIMWT
			of SEQ ID NO: 52
56	chAb6	VH		QVQLQQSGAELMKPGASVKISCKAT GY
				TFSRYWIE WVKQRPGHGLEWIG EILPG
				SGSTNYNEKFKG KATFTADTSSNTAYM
				QVSSLTSEDSAVHYCAR RGYGYVPYAL
				DY WGQGTSVTVSS
57	chAb6	CDR-H1	Residues 26-35
GYTFSRYWIE
			of SEQ ID NO: 56
58	chAb6	CDR-H2	Residues 50-66
EILPGSGSTNYNEKFKG
			of SEQ ID NO: 56
59	chAb6	CDR-H3	Residues 99-110
RGYGYVPYALDY
			of SEQ ID NO: 56
60	chAb6	VL		EIQMTQTTSSLSASLGDRVTISC RASQ
				DISNSLN WYQQKPDGTVNLLIY YTSRL
				YS GVPSRFSGSGSGTDYSLTISNLEQE
				DIATYFC QQGNTLPYT FGGGTKLEIK
61	chAb6	CDR-L1	Residues 24-34
RASQDISNSLN
			of SEQ ID NO: 60
62	chAb6	CDR-L2	Residues 50-56
YTSRLYS
			of SEQ ID NO: 60
63	chAb6	CDR-L3	Residues 89-97
QQGNTLPYT
			of SEQ ID NO: 60
64	chAb11	VH		EVKLVESGGGLVQPGGSLRLSCATS GF
				TFTNYYMS WVRQPPGKALEWLG FIRNK
				ANDYTTEYSASVKG RFTISRDNSQSIL
				YLQMNTLRAEDSATYYCAR ESPGNPFA
				Y WGQGTLVTVSA
65	chAb11	CDR-H1	Residues 26-35
GFTFTNYYMS
			of SEQ ID NO: 64
66	chAb11	CDR-H2	Residues 50-68
FIRNKANDYTTEYSASVKG
			of SEQ ID NO: 64
67	chAb11	CDR-H3	Residues 101-109
ESPGNPFAY
			of SEQ ID NO: 64
68	chAb11	VL		DIVMTQSPSSLTVTAGEKVTMTC KSSQ
				SLLNSGTQKNFLT WYQQKPGQPPKLLI
				Y WASTRES GVPDRFTGSGSGTDFTLTI
				SSVQAEDLAVYFC QNDYIYPLT FGAGT
				KLELK
69	chAb11	CDR-L1	Residues 24-40
KSSQSLLNSGTQKNFLT
			of SEQ ID NO: 68
70	chAb11	CDR-L2	Residues 56-62
WASTRES
			of SEQ ID NO: 68
71	chAb11	CDR-L3	Residues 95-103
QNDYIYPLT
			of SEQ ID NO: 68
72	chAb16	VH		EVKLLESGGGLVQPGGSLKLSCAAS GF
				DFSRYWMS WVRQAPGKGLEWIG EINPD
				SSTINYTPSLKD KFIISRDNAKNTLYL
				QMSKVRSEDTALYYCAR PGFGNYIYAM
				DY WGQGTSVTVSS
73	chAb16	CDR-H1	Residues 26-35
GFDFSRYWMS
			of SEQ ID NO: 72
74	chAb16	CDR-H2	Residues 50-66
EINPDSSTINYTPSLKD
			of SEQ ID NO: 72
75	chAb16	CDR-H3	Residues 99-110
PGFGNYIYAMDY
			of SEQ ID NO: 72
76	chAb16	VL		DIQMTQTTSSLSASLGDRVTINC RASQ
				DISNFLN WYQQKPDGTVKLLIY YTSRL
				YL GVPSRFSGSGSGTDYSLTISNLEQE
				DIATYFC QQGNTLPPT FGGGTKLEIK
77	chAb16	CDR-L1	Residues 24-34
RASQDISNFLN
			of SEQ ID NO: 76
78	chAb16	CDR-L2	Residues 50-56
YTSRLYL
			of SEQ ID NO: 76
79	chAb16	CDR-L3	Residues 89-97
QQGNTLPPT
			of SEQ ID NO: 76
80	chAb10	VH		DVQLQESGPGLVKPSQSLSLTCTVT GY
				SITSDYAWN WIRQFPGNRLEWMG HINY
				SGITNYNPSLKS RISITRDTSKNQFFL
				QLYSVTTEDTATYFCAR RSLFYYYGSS
				LYAMDY WGQGTSVIVSS
81	chAb10	CDR-H1	Residues 26-36
GYSITSDYAWN
			of SEQ ID NO: 80
82	chAb10	CDR-H2	Residues 51-66
HINYSGITNYNPSLKS
			of SEQ ID NO: 80
83	chAb10	CDR-H3	Residues 99-114
RSLFYYYGSSLYAMDY
			of SEQ ID NO: 80
84	chAb10	VL		DVVMTQSPFSLPVSLGDQASISC RSSQ
				SLVHSNGNTYLH WYLQKPGQSPKLLIY
				KVSNRFS GVPDRFSGSGSGTDFTLKIS
				RVEAEDLGVYFC SQSTHVPWT FGGGTK
				LEIK
85	chAb10	CDR-L1	Residues 24-39
RSSQSLVHSNGNTYLH
			of SEQ ID NO: 84
7	chAb10	CDR-L2	Residues 55-61
KVSNRFS
			of SEQ ID NO: 84
86	chAb10	CDR-L3	Residues 94-102
SQSTHVPWT
			of SEQ ID NO: 84
87	chAb7	VH		EVQLVESGENLVKPGGSLKLSCAAS GF
				SFRGYGMS WVRQTPDKRLEWVA AISTG
				GNYTYYPDSVQG RFTISRDNANNTLYL
				QMSSLKSEDTAMYYCAR RGGNYAGFAY
				WGQGTLVTVSA
88	chAb7	CDR-H1	Residues 26-35
GFSFRGYGMS
			of SEQ ID NO: 87
89	chAb7	CDR-H2	Residues 50-66
AISTGGNYTYYPDSVQG
			of SEQ ID NO: 87
90	chAb7	CDR-H3	Residues 99-108
RGGNYAGFAY
			of SEQ ID NO: 87
91	chAb7	VL		DIQMTQSPASLSVSVGETVTITC RPSE
				NIYSNLA WYQQKQGKSPQLLVY AATNL
				AD GVPSRFSGSGSGTQYSLKINSLQSE
				DFGTYYC QHFWGTPFT FGSGTKLEIK
92	chAb7	CDR-L1	Residues 24-34
RPSENIYSNLA
			of SEQ ID NO: 91
93	chAb7	CDR-L2	Residues 50-56
AATNLAD
			of SEQ ID NO: 91
94	chAb7	CDR-L3	Residues 89-97
QHFWGTPFT
			of SEQ ID NO: 91
95	chAb8	VH		EVKLVESGGGLVKPGGSLKLSCAAS GF
				TFSSYGMS WVRQTPEKRLEWVA TISGG
				GNYTYCPDSVKG RFTISRDNAKNNLYL
				QMSSLRSEDTALYYCTR QRGYDYHYAM
				DF WGQGTSVTVSS
49	chAb8	CDR-H1	Residues 26-35
GFTFSSYGMS
			of SEQ ID NO: 95
96	chAb8	CDR-H2	Residues 50-66
TISGGGNYTYCPDSVKG
			of SEQ ID NO: 95
97	chAb8	CDR-H3	Residues 99-110
QRGYDYHYAMDF
			of SEQ ID NO: 95
98	chAb8	VL		DIQMTQSPASLSVSVGETVTITC RASE
				NIYSNLA WHQQKQGKSPQLLVY AATNL
				AD GVPSRFSGNGSDTQYSLKINSLQSE
				DFGSYFC QNFWGTSWT FGGGTKLEIK
99	chAb8	CDR-L1	Residues 24-34
RASENIYSNLA
			of SEQ ID NO: 98
93	chAb8	CDR-L2	Residues 50-56
AATNLAD
			of SEQ ID NO: 98
100	chAb8	CDR-L3	Residues 89-97
QNFWGTSWT
			of SEQ ID NO: 98
101	chAb17	VH		EVKLVESGGGLVQPGGSLKLSCAAS GF
				TFSSYIMS WVRQTPEKRLEWVA SIVSS
				NITYYPDSMKG RFTISRDNARNILYLQ
				MSSLKSEDTAMYYCAR SGTRAWFAY WG
				QGTLVTVSA
102	chAb17	CDR-H1	Residues 26-35
GFTFSSYIMS
			of SEQ ID NO: 101
103	chAb17	CDR-H2	Residues 50-65
SIVSSNITYYPDSMKG
			of SEQ ID NO: 101
104	chAb17	CDR-H3	Residues 98-106
SGTRAWFAY
			of SEQ ID NO: 101
105	chAb17	VL		DIVLTQSPASLAVSLGQRATISC RASK
				SVSTSAYSYMH WYQQKPGQPPKLLIY L
				ASNLES GVPARFSGSGSGTDFTLNIHP
				VEEEDAATYYC QHSRELPYT FGGGTKL
				EIK
106	chAb17	CDR-L1	Residues 24-38
RASKSVSTSAYSYMH
			of SEQ ID NO: 105
46	chAb17	CDR-L2	Residues 54-60
LASNLES
			of SEQ ID NO: 105
107	chAb17	CDR-L3	Residues 93-101
QHSRELPYT
			of SEQ ID NO: 105
108	chAb5	VH		QVQLQQPGDELVKPGASVKLSCKTS GY
				TFTTDWMH WVKQRPGQGLEWIG MIHPN
				SGTTNYNEKFKS KAALTVDKSSSTACM
				QLSSLTSEDSAVYYCAR SYWKWYFDV W
				GTGTTVTVSS
109	chAb5	CDR-H1	Residues 26-35
GYTFTTDWMH
			of SEQ ID NO: 108
110	chAb5	CDR-H2	Residues 50-66
MIHPNSGTTNYNEKFKS
			of SEQ ID NO: 108
111	chAb5	CDR-H3	Residues 99-107
SYWKWYFDV
			of SEQ ID NO: 108
112	chAb5	VL		QIVLTQSPAIMSASLGEEITLTC SASS
				SVSYMH WYQQKSGTSPKLLIY STSNLA
				S GVPSRFSGSGSGTFYSLTISSVEAED
				SADYYC HQWTSYMYT FGGGTKLEIK
113	chAb5	CDR-L1	Residues 24-33
SASSSVSYMH
			of SEQ ID NO: 112
114	chAb5	CDR-L2	Residues 49-55
STSNLAS
			of SEQ ID NO: 112
115	chAb5	CDR-L3	Residues 88-96
HQWTSYMYT
			of SEQ ID NO: 112

TABLE 8 — VH and VL amino acid sequences of humanized versions of chAb18 SEQ

ID		Protein
NO:	Clone	Region	Residues	Amino Acid Sequence
116	huAb18VH.1	VH		EVQLVQSGAEVKKPGSSVKVSCKAS
				GYSFTSYTIH WVRQAPGQGLEWMG Y
				INPNSRNTDYNQKFKD RVTITADKS
				TSTAYMELSSLRSEDTAVYYCAR YS
				GSTPYWYFDV WGQGTTVTVSS
25	huAb18VH.1	CDR-H1	Residues 26-35
GYSFTSYTIH
			of SEQ ID NO: 116
26	huAb18VH.1	CDR-H2	Residues 50-66
YINPNSRNTDYNQKFKD
			of SEQ ID NO: 116
27	huAb18VH.1	CDR-H3	Residues 99-110
YSGSTPYWYFDV
			of SEQ ID NO: 116
117	huAb18VH.1a	VH		EVQLVQSGAEVKKPGSSVKVSCKAS
				GYSFTSYTIH WVRQAPGQGLEWIG Y
				INPNSRNTDYNQKFKD RTTLTADRS
				TSTAYMELSSLRSEDTAVYYCAR YS
				GSTPYWYFDV WGQGTTVTVSS
25	huAb18VH.1a	CDR-H1	Residues 26-35
GYSFTSYTIH
			of SEQ ID NO: 117
26	huAb18VH.1a	CDR-H2	Residues 50-66
YINPNSRNTDYNQKFKD
			of SEQ ID NO: 117
27	huAb18VH.1a	CDR-H3	Residues 99-110
YSGSTPYWYFDV
			of SEQ ID NO: 117
118	huAbl8VH.1b	VH		EVQLVQSGAEVKKPGSSVKVSCKAS
				GYSFTSYTIH WVRQAPGQGLEWMG Y
				INPNSRNTDYAQKFQG RVTLTADKS
				TSTAYMELSSLRSEDTAVYYCAR YS
				GSTPYWYFDV WGQGTTVTVSS
25	huAbl8VH.1b	CDR-H1	Residues 26-35
GYSFTSYTIH
			of SEQ ID NO: 118
119	huAbl8VH.1b	CDR-H2	Residues 50-66
YINPNSRNTDYAQKFQG
			of SEQ ID NO: 118
27	huAbl8VH.1b	CDR-H3	Residues 99-110
YSGSTPYWYFDV
			of SEQ ID NO: 118
120	huAbl8VL.1	VL		DIQLTQSPSFLSASVGDRVTITC RA
				SSSVSYMN WYQQKPGKAPKLLIY AT
				SNLAS GVPSRFSGSGSGTEFTLTIS
				SLQPEDFATYYC QQWSSNPLT FGQG
				TKLEIK
29	huAb18VL.1	CDR-L1	Residues 24-33
RASSSVSYMN
			of SEQ ID NO: 120
30	huAb18VL.1	CDR-L2	Residues 49-55
ATSNLAS
			of SEQ ID NO: 120
31	huAb18VL.1	CDR-L3	Residues 88-96
QQWSSNPLT
			of SEQ ID NO: 120
121	huAb18VL.1a	VL		DIQLTQSPSFLSASVGDRVTITC RA
				SSSVSYMN WYQQKPGKSPKPWIY AT
				SNLAS GVPSRFSVSVSGTEHTLTIS
				SLQPEDFATYYC QQWSSNPLT FGQG
				TKLEIK
29	hu18AbVL.1a	CDR-L1	Residues 24-33
RASSSVSYMN
			of SEQ ID NO: 121
30	huAb18VL.1a	CDR-L2	Residues 49-55
ATSNLAS
			of SEQ ID NO: 121
31	huAb18VL.1a	CDR-L3	Residues 88-96
QQWSSNPLT
			of SEQ ID NO: 121
122	huAb18VL.1b	VL		DIQLTQSPSFLSASVGDRVTITC RA
				SSSVSYMN WYQQKPGKAPKPWIY AT
				SNLAS GVPSRFSVSGSGTEHTLTIS
				SLQPEDFATYYC QQWSSNPLT FGQG
				TKLEIK
29	huAb18VL.1b	CDR-L1	Residues 24-33
RASSSVSYMN
			of SEQ ID NO: 122
30	huAb18VL.1b	CDR-L2	Residues 49-55
ATSNLAS
			of SEQ ID NO: 122
31	huAb18VL.1b	CDR-L3	Residues 88-96
QQWSSNPLT
			of SEQ ID NO: 122
123	huAb18VL.2	VL		EIVLTQSPDFQSVTPKEKVTITC RA
				SSSVSYMN WYQQKPDQSPKLLIK AT
				SNLAS GVPSRFSGSGSGTDFTLTIN
				SLEAEDAATYYC QQWSSNPLT FGQG
				TKLEIK
29	huAb18VL.2	CDR-L1	Residues 24-33
RASSSVSYMN
			of SEQ ID NO: 123
30	huAb18VL.2	CDR-L2	Residues 49-55
ATSNLAS
			of SEQ ID NO: 123
31	huAb18VL.2	CDR-L3	Residues 88-96
QQWSSNPLT
			of SEQ ID NO: 123
124	huAb18VL.2a	VL		EIVLTQSPDFQSVTPKEKVTITC RA
				SSSVSYMN WYQQKPDQSPKPWIY AT
				SNLAS GVPSRFSVSVSGTDHTLTIN
				SLEAEDAATYYC QQWSSNPLT FGQG
				TKLEIK
29	huAb18VL.2a	CDR-L1	Residues 24-33
RASSSVSYMN
			of SEQ ID NO: 124
30	huAbl8VL.2a	CDR-L2	Residues 49-55
ATSNLAS
			of SEQ ID NO: 124
31	huAb18VL.2a	CDR-L3	Residues 88-96
QQWSSNPLT
			of SEQ ID NO: 124

TABLE 9 — Anti-B7-H3 Ab18 humanized antibodies

huAb18v1	huAb18 VH1/huAb18VL1
huAb18v2	huAb18 VH1b/huAb18VL1
huAb18v3	huAb18 VH1a/huAbVL1a
huAb18v4	huAb18 VH1b/huAb18VL1a
huAb18v5	huAn18 VH1/huAb18VL2
huAb18v6	huAb18 VH1b/huAb18VL2
huAb18v7	huAb18 VH1b/huAb18 VL2a
huAb18v8	huAb18 VH1a/huAb18 VL1b
huAb18v9	huAb18 VH1a/huAb18 VL2a
huAb18v10	huAb18 VH1b/huAb18 VL1b

TABLE 10 — In vitro characterization of humanized anti-B7-H3 Ab18 variants Affinity of

				FACS	naked
		DAR		Binding	mAbs	Cytotoxicity
	Sequence	by		to hu B7-	(Biacore,	(HCC38 Cell
Variant name	Numbers	MS	% agg by SEC	H3)	K D )	line IC 50 )
chAb18-CZ	24, 28	2.3		1.14	1.70E−10	0.28
huAb18v1-CZ	116, 120	2.6	3.3	1.27	5.20E−10	0.39
huAb18v2-CZ	118, 120	1.8	3.3	2.25	6.90E−10	1.19
huAb18v3-CZ	117, 121	2.4	3.6	1.27	2.30E−10	0.32
huAb18v4-CZ	118, 121	2.5	5.5	0.90	5.70E−10	0.29
huAb18v5-CZ	116, 123	3.4	4.2	2.91	2.30E−10	0.12
huAb18v6-CZ	118, 123	3.4	3.5	2.09	2.00E−10	0.14
huAb18v7-CZ	118, 124	4.3	3.6	1.92	4.00E−10	0.03
huAb18v8-CZ	117, 122	2.6	3.5	1.98	2.50E−10	1.3
huAb18v9-CZ	117, 124	2.4	3.8	1.58	3.80E−10	0.9
huAb18v10-CZ	118, 122	2.7	3.2	1.19	2.50E−10	0.57

TABLE 11 — In vivo efficacy of anti-B7-H3 ADCS (humanized chAb18-CZ variants)

	Conjugation	DAR by	Dose [a] //route/	Number of	TGI max
ADC	Method	MS	regimen	mice	(%)	TGD (%)
AB095	—	n/a	6 mg/kg/IP/QDx1	8	0	0
huAb18v1-CZ	A	2.6	6 mg/kg/IP/QDx1	8	79	45
huAb18v3-CZ	A	2.4	6 mg/kg/IP/QDx1	8	81	39
huAB18v4-CZ	A	2.5	6 mg/kg/IP/QDx1	8	85	48
huAB18v6-CZ	A	3.4	6 mg/kg/IP/QDx1	8	86	45
huAb18v7-CZ	A	4.3	6 mg/kg/IP/QDx1	8	87	42
huAb18v9-CZ	A	2.4	6 mg/kg/IP/QDx1	8	83	35
[a] dose is given in mg/kg/day

TABLE 12 — VH and VL sequences of humanized versions of chAb3 SEQ

ID		Protein
NO:	Clone	Region	Residues	Amino Acid Sequence
125	huAb3VH.1	VH		EVQLVQSGAEVKKPGSSVKVS
				CKAS GYTFSSYWMH WVRQAPG
				QGLEWMG LIHPDSGSTNYNEM
				FKN RVTITADKSTSTAYMELS
				SLRSEDTAVYYCAR GGRLYFD
				Y WGQGTTVTVSS
10	huAb3VH.1	CDR-H1	Residues 26-35
GYTFSSYWMH
			of SEQ ID NO: 125
11	huAb3VH.1	CDR-H2	Residues 50-66
LIHPDSGSTNYNEMFKR
			of SEQ ID NO: 125
12	huAb3VH.1	CDR-H3	Residues 99-106
GGRLYFDY
			of SEQ ID NO: 125
126	huAb3VH.1a	VH		EVQLVQSGAEVKKPGSSVKVS
				CKAS GYTFSSYWMH WVRQAPG
				QGLEWIG LIHPDSGSTNYNEM
				FKN RATLTVDRSTSTAYVELS
				SLRSEDTAVYFCAG GGRLYFD
				Y WGQGTTVTVSS
10	huAb3VH.1a	CDR-H1	Residues 26-35
GYTFSSYWMH
			of SEQ ID NO: 126
11	huAb3VH.1a	CDR-H2	Residues 50-66
LIHPDSGSTNYNEMFKR
			of SEQ ID NO: 126
12	huAb3VH.1a	CDR-H3	Residues 99-106
GGRLYFDY
			of SEQ ID NO: 126
127	huAb3VH.1b	VH		EVQLVQSGAEVKKPGSSVKVS
				CKAS GYTFSSYWMH WVRQAPG
				QGLEWIG LIHPDSGSTNYNEM
				FKN RATLTVDRSTSTAYMELS
				SLRSEDTAVYYCAG GGRLYFD
				Y WGQGTTVTVSS
10	huAb3VH.1b	CDR-H1	Residues 26-35
GYTFSSYWMH
			of SEQ ID NO: 127
11	huAb3VH.1b	CDR-H2	Residues 50-66
LIHPDSGSTNYNEMFKR
			of SEQ ID NO: 127
12	huAb3VH.1b	CDR-H3	Residues 99-106
GGRLYFDY
			of SEQ ID NO: 127
128	huAb3VL.1	VL		DIVMTQSPLSLPVTPGEPASI
				SC RSSQSLVHSNGDTYLR WYL
				QKPGQSPQLLIY KVSNRFS GV
				PDRFSGSGSGTDFTLKISRVE
				AEDVGVYYC SQSTHVPYT FGG
				GTKVEIK
14	huAb3VL.1	CDR-L1	Residues 24-39
RSSQSLVHSNGDTYLR
			of SEQ ID NO: 128
7	huAb3VL.1	CDR-L2	Residues 55-61
KVSNRFS
			of SEQ ID NO: 128
15	huAb3VL.1	CDR-L3	Residues 94-102
SQSTHVPYT
			of SEQ ID NO: 128
129	huAb3VL.1a	VL		DVVMTQSPLSLPVTPGEPASI
				SC RSSQSLVHSNGDTYLR WYL
				QKPGQSPQLLIY KVSNRFS GV
				PDRFSGSGSGTDFTLKISRVE
				AEDVGVYFC SQSTHVPYT FGG
				GTKVEIK
14	huAb3VL.1a	CDR-L1	Residues 24-39
RSSQSLVHSNGDTYLR
			of SEQ ID NO: 129
7	huAb3VL.1a	CDR-L2	Residues 55-61
KVSNRFS
			of SEQ ID NO: 129
15	huAb3VL.1a	CDR-L3	Residues 94-102
SQSTHVPYT
			of SEQ ID NO: 129
130	huAb3VL.1b	VL		DVVMTQSPLSLPVTPGEPASI
				SC RSSQSLVHSNGDTYLR WYL
				QKPGQSPQLLIY KVSNRFS GV
				PDRFSGSGSGTDFTLKISRVE
				AEDVGVYYC SQSTHVPYT FGG
				GTKVEIK
14	huAb3VL.1b	CDR-L1	Residues 24-39
RSSQSLVHSNGDTYLR
			of SEQ ID NO: 130
7	huAb3VL.1b	CDR-L2	Residues 55-61
KVSNRFS
			of SEQ ID NO: 130
15	huAb3VL.1b	CDR-L3	Residues 94-102
SQSTHVPYT
			of SEQ ID NO: 130

TABLE 13 — Humanized Ab3 antibodies

huAb3v1	huAb3 VH1/huAb3 VL1
huAb3v2	huAb3 VH1b/huAb3 VL1
huAb3v3	huAb3 VH1a/huAb3 VL1a
huAb3v4	huAb3 VH1/huAb3 VL1b
huAb3v5	huAb3 VH1b/huAb3 VL1b
huAb3v6	huAb3 VH1a/huAb3 VL1b

TABLE 14 — In vitro characterization of humanized variants of chAb3

					FACS		Cytotoxicity
			DAR		Binding to	Affinity of	(HCC38 Cell
	Seq. Id.	Conjugation	by	% agg	hu B7-H3)	naked mAbs	line IC 50 )
ADC	Number	Method	MS	by SEC	EC 50 (nM)	(Biacore, K D )	(nM)
chAb3-CZ	9, 13	A	3.8		0.61	1.90E−08	0.17
huAb3v1-	125, 128	A	3.6	3.3	1.45	5.20E−10	0.53
CZ
huAb3v2-	127, 128	A	3.8	10.1	0.73	6.90E−10	0.13
CZ
huAb3v3-	126, 129	A	3.6	2.5	1.68	2.30E−10	9.22
CZ
huAb3v4-	125, 130	A	3.1	3.1	n/a	5.70E−10	n/a
CZ
huAb3v5-	127, 130	A	3.1	5.9	0.85	2.30E−10	0.17
CZ
huAb3v6-	126, 130	A	3.3	4.9	1.78	2.00E−10	0.13
CZ

TABLE 15 — In vivo efficacy of humanized chAb3-CZ variants

	Conjugation		Dose [a] /route/	Number of
ADC	Method	DAR	regimen	mice	TGI max (%)	TGD (%)
AB095	—		6 mg/kg/IP/QDx1	8	0	0
huAb3v2-CZ	A	3.8	6 mg/kg/IP/QDx1	8	83	52
huAb3v6-CZ	A	3.3	6 mg/kg/IP/QDx1	8	91	88
[a] dose is given in mg/kg/day

TABLE 16 — Variable region sequences of huAb3v2 antibody variants SEQ

ID	Clone
NO:	Region	Protein	Residues	Amino Acid Sequence
131	huAb3v2.1	VH		EVQLVQSGAEVKKPGSSVKVSCK
				AS GYTFSSYWMH WVRQAPGQGLE
				WIG LIHPWSGSTNYNEMFKN RAT
				LTVDRSTSTAYMELSSLRSEDTA
				VYYCAG GGRLYFDY WGQGTTVTV
				SS
10	huAb3v2.1	CDR-H1	Residues 26-35
GYTFSSYWMH
			of SEQ ID NO: 131
132	huAb3v2.1	CDR-H2	Residues 50-66
LIHPWSGSTNYNEMFKN
			of SEQ ID NO: 131
12	huAb3v2.1	CDR-H3	Residues 99-106
GGRLYFDY
			of SEQ ID NO: 131
133	huAb3v2.1	VL		DIVMTQSPLSLPVTPGEPASISC
				RSSQSLVHSSGDTYLR WYLQKPG
				QSPQLLIY KVSNRFS GVPDRFSG
				SGSGTDFTLKISRVEAEDVGVYY
				C SQSTHVPYT FGGGTKVEIK
134	huAb3v2.1	CDR-L1	Residues 24-39
RSSQSLVHSSGDTYLR
			of SEQ ID NO: 133
7	huAb3v2.1	CDR-L2	Residues 55-61
KVSNRFS
			of SEQ ID NO: 133
15	huAb3v2.1	CDR-L3	Residues 94-102
SQSTHVPYT
			of SEQ ID NO: 133
131	huAb3v2.2	VH		EVQLVQSGAEVKKPGSSVKVSCK
				AS GYTFSSYWMH WVRQAPGQGLE
				WIG LIHPWSGSTNYNEMFKN RAT
				LTVDRSTSTAYMELSSLRSEDTA
				VYYCAG GGRLYFDY WGQGTTVTV
				SS
10	huAb3v2.2	CDR-H1	Residues 26-35
GYTFSSYWMH
			of SEQ ID NO: 131
132	huAb3v2.2	CDR-H2	Residues 50-66
LIHPWSGSTNYNEMFKN
			of SEQ ID NO: 131
12	huAb3v2.2	CDR-H3	Residues 99-106
GGRLYFDY
			of SEQ ID NO: 131
135	huAb3v2.2	VL		DIVMTQSPLSLPVTPGEPASISC
				RSSQSLVHSNRDTYLR WYLQKPG
				QSPQLLIY KVSNRFS GVPDRFSG
				SGSGTDFTLKISRVEAEDVGVYY
				C SQSTHVPYT FGGGTKVEIK
136	huAb3v2.2	CDR-L1	Residues 24-39
RSSQSLVHSNRDTYLR
			of SEQ ID NO: 135
7	huAb3v2.2	CDR-L2	Residues 55-61
KVSNRFS
			of SEQ ID NO: 135
15	huAb3v2.2	CDR-L3	Residues 94-102
SQSTHVPYT
			of SEQ ID NO: 135
131	huAb3v2.3	VH		EVQLVQSGAEVKKPGSSVKVSCK
				AS GYTFSSYWMH WVRQAPGQGLE
				WIG LIHPWSGSTNYNEMFKN RAT
				LTVDRSTSTAYMELSSLRSEDTA
				VYYCAG GGRLYFDY WGQGTTVTV
				SS
10	huAb3v2.3	CDR-H1	Residues 26-35
GYTFSSYWMH
			of SEQ ID NO: 131
132	huAb3v2.3	CDR-H2	Residues 50-66
LIHPWSGSTNYNEMFKN
			of SEQ ID NO: 131
12	huAb3v2.3	CDR-H3	Residues 99-106
GGRLYFDY
			of SEQ ID NO: 131
137	huAb3v2.3	VL		DIVMTQSPLSLPVTPGEPASISC
				RSSQSLVHSNQDTYLR WYLQKPG
				QSPQLLIY KVSNRFS GVPDRFSG
				SGSGTDFTLKISRVEAEDVGVYY
				C SQSTHVPYT FGGGTKVEIK
138	huAb3v2.3	CDR-L1	Residues 24-39
RSSQSLVHSNQDTYLR
			of SEQ ID NO: 137
7	huAb3v2.3	CDR-L2	Residues 55-61
KVSNRFS
			of SEQ ID NO: 137
15	huAb3v2.3	CDR-L3	Residues 94-102
SQSTHVPYT
			of SEQ ID NO: 137
139	huAb3v2.4	VH		EVQLVQSGAEVKKPGSSVKVSCK
				AS GYTFSSYWMH WVRQAPGQGLE
				WIG LIHPESGSTNYNEMFKN RAT
				LTVDRSTSTAYMELSSLRSEDTA
				VYYCAG GGRLYFDY WGQGTTVTV
				SS
10	huAb3v2.4	CDR-H1	Residues 26-35
GYTFSSYWMH
			of SEQ ID NO: 139
140	huAb3v2.4	CDR-H2	Residues 50-66
LIHPESGSTNYNEMFKN
			of SEQ ID NO: 139
12	huAb3v2.4	CDR-H3	Residues 99-106
GGRLYFDY
			of SEQ ID NO: 139
133	huAb3v2.4	VL		DIVMTQSPLSLPVTPGEPASISC
				RSSQSLVHSSGDTYLR WYLQKPG
				QSPQLLIY KVSNRFS GVPDRFSG
				SGSGTDFTLKISRVEAEDVGVYY
				C SQSTHVPYT FGGGTKVEIK
134	huAb3v2.4	CDR-L1	Residues 24-39
RSSQSLVHSSGDTYLR
			of SEQ ID NO: 133
7	huAb3v2.4	CDR-L2	Residues 55-61
KVSNRFS
			of SEQ ID NO: 133
15	huAb3v2.4	CDR-L3	Residues 94-102
SQSTHVPYT
			of SEQ ID NO: 133
139	huAb3v2.5	VH		EVQLVQSGAEVKKPGSSVKVSCK
				AS GYTFSSYWMH WVRQAPGQGLE
				WIG LIHPESGSTNYNEMFKN RAT
				LTVDRSTSTAYMELSSLRSEDTA
				VYYCAG GGRLYFDY WGQGTTVTV
				SS
10	huAb3v2.5	CDR-H1	Residues 26-35
GYTFSSYWMH
			of SEQ ID NO: 139
140	huAb3v2.5	CDR-H2	Residues 50-66
LIHPESGSTNYNEMFKN
			of SEQ ID NO: 139
12	huAb3v2.5	CDR-H3	Residues 99-106
GGRLYFDY
			of SEQ ID NO: 139
135	huAb3v2.5	VL		DIVMTQSPLSLPVTPGEPASISC
				RSSQSLVHSNRDTYLR WYLQKPG
				QSPQLLIY KVSNRFS GVPDRFSG
				SGSGTDFTLKISRVEAEDVGVYY
				C SQSTHVPYT FGGGTKVEIK
136	huAb3v2.5	CDR-L1	Residues 24-39
RSSQSLVHSNRDTYLR
			of SEQ ID NO: 135
7	huAb3v2.5	CDR-L2	Residues 55-61
KVSNRFS
			of SEQ ID NO: 135
15	huAb3v2.5	CDR-L3	Residues 94-102
SQSTHVPYT
			of SEQ ID NO: 135
139	huAb3v2.6	VH		EVQLVQSGAEVKKPGSSVKVSCK
				AS GYTFSSYWMH WVRQAPGQGLE
				WIG LIHPESGSTNYNEMFKN RAT
				LTVDRSTSTAYMELSSLRSEDTA
				VYYCAG GGRLYFDY WGQGTTVTV
				SS
10	huAb3v2.6	CDR-H1	Residues 26-35
GYTFSSYWMH
			of SEQ ID NO: 139
140	huAb3v2.6	CDR-H2	Residues 50-66
LIHPESGSTNYNEMFKN
			of SEQ ID NO: 139
12	huAb3v2.6	CDR-H3	Residues 99-106
GGRLYFDY
			of SEQ ID NO: 139
137	huAb3v2.6	VL		DIVMTQSPLSLPVTPGEPASISC
				RSSQSLVHSNQDTYLR WYLQKPG
				QSPQLLIY KVSNRFS GVPDRFSG
				SGSGTDFTLKISRVEAEDVGVYY
				C SQSTHVPYT FGGGTKVEIK
138	huAb3v2.6	CDR-L1	Residues 24-39
RSSQSLVHSNQDTYLR
			of SEQ ID NO: 137
7	huAb3v2.6	CDR-L2	Residues 55-61
KVSNRFS
			of SEQ ID NO: 137
15	huAb3v2.6	CDR-L3	Residues 94-102
SQSTHVPYT
			of SEQ ID NO: 137
141	huAb3v2.7			EVQLVQSGAEVKKPGSSVKVSCK
				AS GYTFSSYWMH WVRQAPGQGLE
				WIG LIHPISGSTNYNEMFKN RAT
				LTVDRSTSTAYMELSSLRSEDTA
				VYYCAG GGRLYFDY WGQGTTVTV
				SS
10	huAb3v2.7	CDR-H1	Residues 26-35
GYTFSSYWMH
			of SEQ ID NO: 141
142	huAb3v2.7	CDR-H2	Residues 50-66
LIHPISGSTNYNEMFKN
			of SEQ ID NO: 141
12	huAb3v2.7	CDR-H3	Residues 99-106
GGRLYFDY
			of SEQ ID NO: 141
133	huAb3v2.7			DIVMTQSPLSLPVTPGEPASISC
				RSSQSLVHSSGDTYLR WYLQKPG
				QSPQLLIY KVSNRFS GVPDRFSG
				SGSGTDFTLKISRVEAEDVGVYY
				C SQSTHVPYT FGGGTKVEIK
134	huAb3v2.7	CDR-L1	Residues 24-39
RSSQSLVHSSGDTYLR
			of SEQ ID NO: 133
7	huAb3v2.7	CDR-L2	Residues 55-61
KVSNRFS
			of SEQ ID NO: 133
15	huAb3v2.7	CDR-L3	Residues 94-102
SQSTHVPYT
			of SEQ ID NO: 133
141	huAb3v2.8	VH		EVQLVQSGAEVKKPGSSVKVSCK
				AS GYTFSSYWMH WVRQAPGQGLE
				WIG LIHPISGSTNYNEMFKN RAT
				LTVDRSTSTAYMELSSLRSEDTA
				VYYCAG GGRLYFDY WGQGTTVTV
				SS
10	huAb3v2.8	CDR-H1	Residues 26-35
GYTFSSYWMH
			of SEQ ID NO: 141
142	huAb3v2.8	CDR-H2	Residues 50-66
LIHPISGSTNYNEMFKN
			of SEQ ID NO: 141
12	huAb3v2.8	CDR-H3	Residues 99-106
GGRLYFDY
			of SEQ ID NO: 141
135	huAb3v2.8	VL		DIVMTQSPLSLPVTPGEPASISC
				RSSQSLVHSNRDTYLR WYLQKPG
				QSPQLLIY KVSNRFS GVPDRFSG
				SGSGTDFTLKISRVEAEDVGVYY
				C SQSTHVPYT FGGGTKVEIK
136	huAb3v2.8	CDR-L1	Residues 24-39
RSSQSLVHSNRDTYLR
			of SEQ ID NO: 135
7	huAb3v2.8	CDR-L2	Residues 55-61
KVSNRFS
			of SEQ ID NO: 135
15	huAb3v2.8	CDR-L3	Residues 94-102
SQSTHVPYT
			of SEQ ID NO: 135
141	huAb3v2.9	VH		EVQLVQSGAEVKKPGSSVKVSCK
				AS GYTFSSYWMH WVRQAPGQGLE
				WIG LIHPISGSTNYNEMFKN RAT
				LTVDRSTSTAYMELSSLRSEDTA
				VYYCAG GGRLYFDY WGQGTTVTV
				SS
10	huAb3v2.9	CDR-H1	Residues 26-35
GYTFSSYWME
			of SEQ ID NO: 141
142	huAb3v2.9	CDR-H2	Residues 50-66
LIHPISGSTNYNEMFKN
			of SEQ ID NO: 141
12	huAb3v2.9	CDR-H3	Residues 99-106
GGRLYFDY
			of SEQ ID NO: 141
137	huAb3v2.9	VL		DIVMTQSPLSLPVTPGEPASISC
				RSSQSLVHSNQDTYLR WYLQKPG
				QSPQLLIY KVSNRFS GVPDRFSG
				SGSGTDFTLKISRVEAEDVGVYY
				C SQSTHVPYT FGGGTKVEIK
138	huAb3v2.9	CDR-L1	Residues 24-39
RSSQSLVHSNQDTYLR
			of SEQ ID NO: 137
7	huAb3v2.9	CDR-L2	Residues 55-61
KVSNRFS
			of SEQ ID NO: 137
15	huAb3v2.9	CDR-L3	Residues 94-102
SQSTHVPYT
			of SEQ ID NO: 137

TABLE 17 — In vitro characterization of humanized huAb3v2 variants, including naked antibodies and ADCs Affinity of

	%		FACS (EC 50	naked	Cytotoxicity
	agg	ELISA	nM)	mAbs	(H847 Cell
	Sequence	Conjugation	DAR	by	hB7-H3	hB7-	cyB7-	(Biacore,	line IC 50 )
ADC	number	Method	by MS	SEC	EC 50 nM	H3	H3	K D )	(nM)
huAb3v2-	127, 128	A	3.5		0.44	5.11	2.87	2.30E−09	1.49
CZ
huAb3v2.2-	131, 135	A	0.7	1.8	0.10	5.29	3.68	Poor fit	26.7
CZ
huAb3v2.3-	131, 137	A	1.1	1.5	0.11	6.50	4.03	Poor fit	—
CZ
huAb3v2.5-	139, 135	A	3.4	15.6	0.13	5.14	4.86	5.30E−09	1.57
CZ
huAb3v2.6-	139, 137	A	3.3	15	0.09	5.64	3.31	5.80E−08	1.70
CZ
huAb3v2.8-	141, 135	A	2.0	5.7	0.14	3.94	3.01	Poor fit	2.36
CZ
huAb3v2.9-	141, 137	A	2.7	4.3	0.16	6.16	4.64	Poor fit	2.30
CZ

TABLE 18 — Amino acid variable region sequences of humanized Ab13 SEQ

ID		Protein
NO:	Clone	Region	Residues	Amino Acid Sequence
143	huAb13VL.1	VL		DIQMTQSPSSLSASVGDRVTIT
				C KASQNVGFNVA WYQQKPGKAP
				KLLIY SASYRYS GVPSRFSGSG
				SGTDFTLTISSLQPEDFATYYC
				QQYNWYPFT FGQGTKLEIK
37	huAb13VL.1	CDR-L1	Residues 24-34
KASQNVGFNVA
			of SEQ ID NO: 143
38	huAb13VL.1	CDR-L2	Residues 50-56
SASYRYS
			of SEQ ID NO: 143
39	huAb13VL.1	CDR-L3	Residues 89-97
QQYNWYPFT
			of SEQ ID NO: 143
144	huAb13VL.1a	VL		DIQMTQSPSSLSASVGDRVTIT
				C KASQNVGFNVA WYQQKPGKSP
				KALIY SASYRYS GVPSRFSGSG
				SGTDFTLTISSLQPEDFAEYFC
				QQYNWYPFT FGQGTKLEIK
37	huAb13VL.1a	CDR-L1	Residues 24-34
KASQNVGFNVA
			of SEQ ID NO: 144
38	huAb13VL.1a	CDR-L2	Residues 50-56
SASYRYS
			of SEQ ID NO: 144
39	huAb13VL.1a	CDR-L3	Residues 89-97
QQYNWYPFT
			of SEQ ID NO: 144
146	huAb13VH.1	VH		EVQLQESGPGLVKPSETLSLTC
				AVS GYSITSGYSWH WIRQPPGK
				GLEWIG YIHSSGSTNYNPSLKS
				RVTISVDTSKNQFSLKLSSVTA
				ADTAVYYCAR YDDYFEY WGQGT
				TVTVSS
33	huAb13VH.1	CDR-H1	Residues 26-36
GYSITSGYSWH
			of SEQ ID NO: 146
34	huAb13VH.1	CDR-H2	Residues 51-66
YIHSSGSTNYNPSLKS
			of SEQ ID NO: 146
35	huAb13VH.1	CDR-H3	Residues 99-105
YDDYFEY
			of SEQ ID NO: 146
145	huAB13VL.1b	VL		DIQMTQSPSSLSASVGDRVTIT
				C KASQNVGFNVA WYQQKPGKAP
				KLLIY SASYRYS GVPSRFSGSG
				SGTDFTLTISSLQPEDFATYFC
				QQYNWYPFT FGQGTKLEIK
37	huAb13VL.1b	CDR-L1	Residues 24-34
KASQNVGFNVA
			of SEQ ID NO: 145
38	huAb13VL.1b	CDR-L2	Residues 50-56
SASYRYS
			of SEQ ID NO: 145
39	huAb13VL.1b	CDR-L3	Residues 89-97
QQYNWYPFT
			of SEQ ID NO: 145
147	huAb13VH.1a	VH		EVQLQESGPGLVKPSETLSLTC
				AVT GYSITSGYSWH WIRQFPGN
				GLEWMG YIHSSGSTNYNPSLKS
				RISISRDTSKNQFFLKLSSVTA
				ADTAVYYCAG YDDYFEY WGQGT
				TVTVSS
33	huAb13VH.1a	CDR-H1	Residues 26-36
GYSITSGYSWH
			of SEQ ID NO: 147
34	huAb13VH.1a	CDR-H2	Residues 51-66
YIHSSGSTNYNPSLKS
			of SEQ ID NO: 147
35	huAb13VH.1a	CDR-H3	Residues 99-105
YDDYFEY
			of SEQ ID NO: 147
148	huAb13VH.1b	VH		EVQLQESGPGLVKPSETLSLTC
				AVS GYSITSGYSWH WIRQPPGN
				GLEWMG YIHSSGSTNYNPSLKS
				RITISRDTSKNQFSLKLSSVTA
				ADTAVYYCAR YDDYFEY WGQGT
				TVTVSS
33	huAb13VH.1b	CDR-H1	Residues 26-36
GYSITSGYSWH
			of SEQ ID NO: 148
34	huAb13VH.1b	CDR-H2	Residues 51-66
YIHSSGSTNYNPSLKS
			of SEQ ID NO: 148
35	huAb13VH.1b	CDR-H3	Residues 99-105
YDDYFEY
			of SEQ ID NO: 148

TABLE 19 — Variable region sequences of engineered huAb13 variants SEQ

ID		Protein
NO:	Clone	Region	Residues	Amino acid sequence
147	huAb13v1	VH		EVQLQESGPGLVKPSETLSLTCAV
				T GYSITSGYSWH WIRQFPGNGLEW
				MG YIHSSGSTNYNPSLKS RISISR
				DTSKNQFFLKLSSVTAADTAVYYC
				AG YDDYFEY WGQGTTVIVSS
33	huAb13v1	CDR-H1	Residues 26-36
GYSITSGYSWH
			of SEQ ID NO: 147
34	huAb13v1	CDR-H2	Residues 51-66
YIHSSGSTNYNPSLKS
			of SEQ ID NO: 147
35	huAb13v1	CDR-H3	Residues 99-105
YDDYFEY
			of SEQ ID NO: 147
144	huAb13v1	VL		DIQMTQSPSSLSASVGDRVTITC K
				ASQNVGFNVA WYQQKPGKSPKALI
				Y SASYRYS GVPSRFSGSGSGTDFT
				LTISSLQPEDFAEYFC QQYNWYPF
				T FGQGTKLEIK
37	huAb13v1	CDR-L1	Residues 24-34
KASQNVGFNVA
			of SEQ ID NO: 144
38	huAb13v1	CDR-L2	Residues 50-56
SASYRYS
			of SEQ ID NO: 144
39	huAb13v1	CDR-L3	Residues 89-97
QQYNWYPFT
			of SEQ ID NO: 144
146	huAb13v2	VH		EVQLQESGPGLVKPSETLSLTCAV
				S GYSITSGYSWH WIRQPPGKGLEW
				IG YIHSSGSTNYNPSLKS RVTISV
				DTSKNQFSLKLSSVTAADTAVYYC
				AR YDDYFEY WGQGTTVIVSS
33	huAb13v2	CDR-H1	Residues 26-36
GYSITSGYSWH
			of SEQ ID NO: 146
34	huAb13v2	CDR-H2	Residues 51-66
YIHSSGSTNYNPSLKS
			of SEQ ID NO: 146
35	huAb13v2	CDR-H3	Residues 99-105
YDDYFEY
			of SEQ ID NO: 146
143	huAb13v2	VL		DIQMTQSPSSLSASVGDRVTITC K
				ASQNVGFNVA WYQQKPGKAPKLLI
				Y SASYRYS GVPSRFSGSGSGTDFT
				LTISSLQPEDFATYYC QQYNWYPF
				T FGQGTKLEIK
37	huAb13v2	CDR-L1	Residues 24-34
KASQNVGFNVA
			of SEQ ID NO: 143
38	huAb13v2	CDR-L2	Residues 50-56
SASYRYS
			of SEQ ID NO: 143
39	huAb13v2	CDR-L3	Residues 89-97
QQYNWYPFT
			of SEQ ID NO: 143
146	huAb13v3	VH		EVQLQESGPGLVKPSETLSLTCAV
				S GYSITSGYSWH WIRQPPGKGLEW
				IG YIHSSGSTNYNPSLKS RVTISV
				DTSKNQFSLKLSSVTAADTAVYYC
				AR YDDYFEY WGQGTTVIVSS
33	huAb13v3	CDR-H1	Residues 26-36
GYSITSGYSWH
			of SEQ ID NO: 146
34	huAb13v3	CDR-H2	Residues 51-66
YIHSSGSTNYNPSLKS
			of SEQ ID NO: 146
35	huAb13v3	CDR-H3	Residues 99-105
YDDYFEY
			of SEQ ID NO: 146
144	huAb13v3	VL		DIQMTQSPSSLSASVGDRVTITC K
				ASQNVGFNVA WYQQKPGKSPKALI
				Y SASYRYS GVPSRFSGSGSGTDFT
				LTISSLQPEDFAEYFC QQYNWYPF
				T FGQGTKLEIK
37	huAb13v3	CDR-L1	Residues 24-34
KASQNVGFNVA
			of SEQ ID NO: 144
38	huAb13v3	CDR-L2	Residues 50-56
SASYRYS
			of SEQ ID NO: 144
39	huAb13v3	CDR-L3	Residues 89-97
QQYNWYPFT
			of SEQ ID NO: 144
146	huAb13v4	VH		EVQLQESGPGLVKPSETLSLTCAV
				S GYSITSGYSWH WIRQPPGKGLEW
				IG YIHSSGSTNYNPSLKS RVTISV
				DTSKNQFSLKLSSVTAADTAVYYC
				AR YDDYFEY WGQGTTVIVSS
33	huAb13v4	CDR-H1	Residues 26-36
GYSITSGYSWH
			of SEQ ID NO: 146
34	huAb13v4	CDR-H2	Residues 51-66
YIHSSGSTNYNPSLKS
			of SEQ ID NO: 146
35	huAb13v4	CDR-H3	Residues 99-105
YDDYFEY
			of SEQ ID NO: 146
145	huAb13v4	VL		DIQMTQSPSSLSASVGDRVTITC K
				ASQNVGFNVA WYQQKPGKAPKLLI
				Y SASYRYS GVPSRFSGSGSGTDFT
				LTISSLQPEDFATYFC QQYNWYPF
				T FGQGTKLEIK
37	huAb13v4	CDR-L1	Residues 24-34
KASQNVGFNVA
			of SEQ ID NO: 145
38	huAb13v4	CDR-L2	Residues 50-56
SASYRYS
			of SEQ ID NO: 145
39	huAb13v4	CDR-L3	Residues 89-97
QQYNWYPFT
			of SEQ ID NO: 145
147	huAb13v5	VH		EVQLQESGPGLVKPSETLSLTCAV
				T GYSITSGYSWH WIRQFPGNGLEW
				MG YIHSSGSTNYNPSLKS RISISR
				DTSKNQFFLKLSSVTAADTAVYYC
				AG YDDYFEY WGQGTTVIVSS
33	huAb13v5	CDR-H1	Residues 26-36
GYSITSGYSWH
			of SEQ ID NO: 147
34	huAb13v5	CDR-H2	Residues 51-66
YIHSSGSTNYNPSLKS
			of SEQ ID NO: 147
35	huAb13v5	CDR-H3	Residues 99-105
YDDYFEY
			of SEQ ID NO: 147
143	huAb13v5	VL		DIQMTQSPSSLSASVGDRVTITC K
				ASQNVGFNVA WYQQKPGKAPKLLI
				Y SASYRYS GVPSRFSGSGSGTDFT
				LTISSLQPEDFATYYC QQYNWYPF
				T FGQGTKLEIK
37	huAb13v5	CDR-L1	Residues 24-34
KASQNVGFNVA
			of SEQ ID NO: 143
38	huAb13v5	CDR-L2	Residues 50-56
SASYRYS
			of SEQ ID NO: 143
39	huAb13v5	CDR-L3	Residues 89-97
QQYNWYPFT
			of SEQ ID NO: 143
147	huAb13v6	VH		EVQLQESGPGLVKPSETLSLTCAV
				T GYSITSGYSWH WIRQFPGNGLEW
				MG YIHSSGSTNYNPSLKS RISISR
				DTSKNQFFLKLSSVTAADTAVYYC
				AG YDDYFEY WGQGTTVIVSS
33	huAb13v6	CDR-H1	Residues 26-36
GYSITSGYSWH
			of SEQ ID NO: 147
34	huAb13v6	CDR-H2	Residues 51-66
YIHSSGSTNYNPSLKS
			of SEQ ID NO: 147
35	huAb13v6	CDR-H3	Residues 99-105
YDDYFEY
			of SEQ ID NO: 147
145	huAb13v6	VL		DIQMTQSPSSLSASVGDRVTITC K
				ASQNVGFNVA WYQQKPGKAPKLLI
				Y SASYRYS GVPSRFSGSGSGTDFT
				LTISSLQPEDFATYFC QQYNWYPF
				T FGQGTKLEIK
37	huAb13v6	CDR-L1	Residues 24-34
KASQNVGFNVA
			of SEQ ID NO: 145
38	huAb13v6	CDR-L2	Residues 50-56
SASYRYS
			of SEQ ID NO: 145
39	huAb13v6	CDR-L3	Residues 89-97
QQYNWYPFT
			of SEQ ID NO: 145
148	huAb13v7	VH		EVQLQESGPGLVKPSETLSLTCAV
				S GYSITSGYSWH WIRQPPGNGLEW
				MG YIHSSGSTNYNPSLKS RITISR
				DTSKNQFSLKLSSVTAADTAVYYC
				AR YDDYFEY WGQGTTVIVSS
33	huAb13v7	CDR-H1	Residues 26-36
GYSITSGYSWH
			of SEQ ID NO: 148
34	huAb13v7	CDR-H2	Residues 51-66
YIHSSGSTNYNPSLKS
			of SEQ ID NO: 148
35	huAb13v7	CDR-H3	Residues 99-105
YDDYFEY
			of SEQ ID NO: 148
143	huAb13v7	VL		DIQMTQSPSSLSASVGDRVTITC K
				ASQNVGFNVA WYQQKPGKAPKLLI
				Y SASYRYS GVPSRFSGSGSGTDFT
				LTISSLQPEDFATYYC QQYNWYPF
				T FGQGTKLEIK
37	huAb13v7	CDR-L1	Residues 24-34
KASQNVGFNVA
			of SEQ ID NO: 143
38	huAb13v7	CDR-L2	Residues 50-56
SASYRYS
			of SEQ ID NO: 143
39	huAb13v7	CDR-L3	Residues 89-97
QQYNWYPFT
			of SEQ ID NO: 143
148	huAb13v8	VH		EVQLQESGPGLVKPSETLSLTCAV
				S GYSITSGYSWH WIRQPPGNGLEW
				MG YIHSSGSTNYNPSLKS RITISR
				DTSKNQFSLKLSSVTAADTAVYYC
				AR YDDYFEY WGQGTTVTVSS
33	huAb13v8	CDR-Hl	Residues 26-36
GYSITSGYSWH
			of SEQ ID NO: 148
34	huAb13v8	CDR-H2	Residues 51-66
YIHSSGSTNYNPSLKS
			of SEQ ID NO: 148
35	huAb13v8	CDR-H3	Residues 99-105
YDDYFEY
			of SEQ ID NO: 148
144	huAb13v8	VL		DIQMTQSPSSLSASVGDRVTITC K
				ASQNVGFNVA WYQQKPGKSPKALI
				Y SASYRYS GVPSRFSGSGSGTDFT
				LTISSLQPEDFAEYFC QQYNWYPF
				T FGQGTKLEIK
37	huAb13v8	CDR-L1	Residues 24-34
KASQNVGFNVA
			of SEQ ID NO: 144
38	huAb13v8	CDR-L2	Residues 50-56
SASYRYS
			of SEQ ID NO: 144
39	huAb13v8	CDR-L3	Residues 89-97
QQYNWYPFT
			of SEQ ID NO: 144
148	huAb13v9	VH		EVQLQESGPGLVKPSETLSLTCAV
				S GYSITSGYSWH WIRQPPGNGLEW
				MG YIHSSGSTNYNPSLKS RITISR
				DTSKNQFSLKLSSVTAADTAVYYC
				AR YDDYFEY WGQGTTVTVSS
33	huAb13v9	CDR-H1	Residues 26-36
GYSITSGYSWH
			of SEQ ID NO: 148
34	huAb13v9	CDR-H2	Residues 51-66
YIHSSGSTNYNPSLKS
			of SEQ ID NO: 148
35	huAb13v9	CDR-H3	Residues 99-105
YDDYFEY
			of SEQ ID NO: 148
145	huAb13v9	VL		DIQMTQSPSSLSASVGDRVTITC K
				ASQNVGFNVA WYQQKPGKAPKLLI
				Y SASYRYS GVPSRFSGSGSGTDFT
				LTISSLQPEDFATYFC QQYNWYPF
				T FGQGTKLEIK
37	huAb13v9	CDR-L1	Residues 24-34
KASQNVGFNVA
			of SEQ ID NO: 145
38	huAb13v9	CDR-L2	Residues 50-56
SASYRYS
			of SEQ ID NO: 145
39	huAb13v9	CDR-L3	Residues 89-97
QQYNWYPFT
			of SEQ ID NO: 145

TABLE 20 — In vitro characterization of huAb13 VL.1a variants conjugated to synthon (or linker payload) CZ

			ELISA		Affinity of
		%	hB7-	FACS (EC 50	naked	Cytotoxicity
	DAR	agg	H3	nM)	mAbs	(H847 Cell
Variant	Sequence	by	by	EC 50	hB7-	cyB7-	(Biacore,	line IC 50 )
name	Number	MS	SEC	nM	H3	H3	K D )	(nM)
chAb13-CZ	32, 36	—	—	0.26	6.27	18.35	5.7E−09	—
huAb13v1-	147, 144	4.0	5.1	0.12	6.01	10.0	6.2E−09	0.09
CZ
huAb13v5-	147, 143	3.4	2.4	0.19	5.21	10.59	Poor fit	1.60
CZ
huAb13v6-	147, 145	3.6	7.3	0.14	5.83	12.95	Poor fit	0.84
CZ

TABLE 21 — In vitro tumor cell cytotoxicity of selected humanized B7-H3 ADCs with exemplary Bcl-xL inhibitor linker warheads (synthons)

			% agg	ADC
	Conjugation	DAR	by	conc	EC 50 nM
ADC	Method	by MS	SEC	(mg/ml)	H1650
huAb13v1-CZ	G	4	3.9	3	0.18
huAb13v1-TX	G	3.6	2.8	2.6	0.22
huAb13v1-TV	G	2.4	3	3.9	0.43
huAb13v1-AAA	G	2	20.2	2.7	0.37
huAb13v1-AAD	G	3.7	3.3	2.7	0.21
huAb13v1-WD	E	3	5.4	5.8	0.45
huAb13v1-LB	A	2.2	21.9	3.7	>133
huAb13v1-ZT	G	2.4	10.6	1.7	0.3
huAb13v1-ZZ	G	1.4	20.3	2.5	0.42
huAb13v1-XW	G	4.3	6.3	2.6	0.86
huAb13v1-SE	A	3.7	4	5.4	0.63
huAb13v1-SR	A	2.6	49.5	4.5	0.59
huAb13v1-YG	E	3.3	2.1	3.8	0.33
huAb13v1-KZ	A	2.8	16.8	3.5	178.8
huAb3v2.5-CZ	G	3.3	15.6	3.6	0.40
huAb3v2.5-TX	G	3.3	8.9	2.9	0.62
huAb3v2.5-TV	G	3.7	10.4	3.5	0.53
huAb3v2.5-YY	G	2.3	16.2	3.2	0.71
huAb3v2.5-AAA	G	2	14.8	3.3	0.85
huAb3v2.5-AAD	G	3.4	11.3	3.7	0.49
huAb3v2.5-WD	E	2.8	11.5	5.4	0.83
huAb3v2.5-LB	A	2.2	24.4	3.9	2.59
huAb3v2.5-ZT	G	1.6	70.1	3.3	0.95
huAb3v2.5-ZZ	G	1.2	19.4	3.7	1.1
huAb3v2.5-XW	G	3.9	16.4	3.4	2.18
huAb3v2.5-SE	A	3.7	10.6	5.4	0.85
huAb3v2.5-SR	A	1.8	48.5	5.1	0.59
huAb3v2.5-YG	E	4	8.6	3.3	0.71
huAb3v2.5-KZ	A	2.6	24.5	3.4	0.87
huAb3v2.6-CZ	G	3.4	15	3.6	0.40
huAb3v2.6-TX	G	3.2	10.4	3.4	0.47
huAb3v2.6-TV	G	3.3	10.7	3.8	0.52
huAb3v2.6-YY	G	2.2	19.9	3.4	0.72
huAb3v2.6-AAA	G	1.9	20.2	3.6	1.24
huAb3v2.6-AAD	G	3.4	11.9	3.7	0.85
huAb3v2.6-WD	E	3.1	12.4	5.3	0.79
huAb3v2.6-LB	A	2.4	27.2	3.9	2.07
huAb3v2.6-ZT	G	1.7	21.6	3.7	1.11
huAb3v2.6-ZZ	G	1.2	70.7	3.5	1.35
huAb3v2.6-XW	G	4	16.8	3.2	2.4
huAb3v2.6-SE	A	3.6	11.8	5.7	1.01
huAb3v2.6-SR	A	2.5	48.2	5.2	0.71
huAb3v2.6-YG	E	3.7	9.9	4.8	0.68
huAb3v2.6-KZ	A	3.5	26.1	3.6	5.52
MSL109-CZ	G	3.2	0.5	3.7	19.50
MSL109-TX	G	3.5	0.7	3	20.00
MSL109-TV	G	3.6	0	2.6	31.13
MSL109-YY	G	2.9	0	1.8	26.53
MSL109-AAA	G	1.9	13.7	3.2	23.52
MSL109-AAD	G	3	0.4	3.8	>67
MSL109-WD	E	2.9	0	7.06	18.22
MSL109-LB	A	1.8	0	4.2	9.88
MSL109-ZT	G	2.3	7.5	2.2	>67
MSL109-ZZ	G	1.4	15	3.5	>67
MSL109-XW	G	3.3	3.7	3.2	>67
MSL109-SE	A	3.6	33.4	6.0	29.56
MSL109-SR	A	1.8	2.3	3.8	53.29
MSL109-YG	E	3.1	13.2	4.0	19.93
MSL109-KZ	A	2.5	18	4.3	50.16

TABLE 22 — In vitro tumor cell cytotoxicity of selected humanized B7-H3 ADCs with exemplary Bcl-xL inhibitor synthons. ADC

	Conjugation		% agg by	conc	EC 50 nM
ADC	Method	DAR	SEC	(mg/ml)	H146
huAb13v1-AAA	I	2	3.3	11.6	2
E2
huAb13v1-WD	I	2	4.5	14.5	2
E2

TABLE 23 — In vivo efficacy of humanized anti-B7-H3 ADCs

	Conjugation		Dose [a] /route/	Number	TGI max
ADC	Method	DAR	regimen	of mice	(%)	TGD (%)
AB095	—	n/a	6 mg/kg/IP/QDx1	8	0	0
huAb3v2.5-CZ	A	3.5	6 mg/kg/IP/QDx1	8	92	122
huAb3v2.6-CZ	A	3.4	6 mg/kg/IP/QDx1	8	93	130
huAb3v2.9-CZ	A	2.8	6 mg/kg/IP/QDx1	8	94	135
huAb3v2.9-TX	E	1.7	6 mg/kg/IP/QDx1	8	93	109
huAb3v2.6-TX	E	2.7	6 mg/kg/IP/QDx1	8	92	130
huAb3v2.5-TX	E	2.5	6 mg/kg/IP/QDx1	8	86	89
[a] dose is given in mg/kg/day

TABLE 24 — In vivo efficacy of humanized anti-B7-H3 ADCs

	Conju-			Number
	gation		Dose [a] /route/	of	TGI max
ADC	Method	DAR	regimen	mice	(%)
AB095	—	n/a	6 mg/kg/IP/QDx1	8	0
huAb3v2.5-AAA	E	2.3	6 mg/kg/IP/QDx1	8	65
huAb3v2.5-XW	E	3.1	6 mg/kg/IP/QDx1	8	51
huAb3v2.6-AAA	E	3.5	6 mg/kg/IP/QDx1	8	47
huAb3v2.6-XW	E	4.0	6 mg/kg/IP/QDx1	8	43
huAb13v1-AAA	E	3.5	6 mg/kg/IP/QDx1	8	76
huAb13v1-XW	E	4.2	6 mg/kg/IP/QDx1	8	35
huAb13v1-TX E2	I	2	6 mg/kg/IP/QDx1	8	88
[a] dose is given in mg/kg/day

TABLE 25 — In vivo efficacy of humanized anti-B7-H3 ADC huAb13v1-WD in H1650

	DAR/	Dose
	Conjugation	mg/kg/	route/	TGI max	TGD
ADC	Method	day	regimen	(%)	(%)
AB095 (a)	N.A.	10	IP/QDx1	0	0
huAb13v1-WD-E2	2/I	1	IP/QDx1	46*	47*
huAb13v1-WD-E2	2/I	3	IP/QDx1	48*	47*
huAb13v1-WD-E2	2/I	10	IP/QDx1	62*	77*
(a) IgG1 mAb
*= p < 0.05 as compared to control treatment (AB095)
¥ = p < 0.05 as compared to the most active partner in a drug combination
N.A. = not applicable

TABLE 26 — In vivo efficacy of non-targeting (MSL109) BCL-xL inhibiting ADCs in NCI-H1650 model of NSCLC Growth Inhibition

		TGI max	TGD
Treatment	Dose [a] /route/regimen	(%)	(%)
MSL109 † -H	3/IP/Q4Dx6	18*	0
MSL109 † -H	10/IP/Q4Dx6	43*	20*
MSL109 † -H	30/IP/Q4Dx6	8	0
MSL109 † -CZ	3/IP/Q4Dx6	29*	0
MSL109 † -CZ	3/IP/Q7Dx6	18*	0
MSL109 † -CZ	10/IP/Q4Dx6	32*	16
MSL109 † -CZ	30/IP/Q4Dx6	32*	12
† Non-targeting antibody
[a] dose is given in mg/kg/day
*= p < 0.05 as compared to control treatment (AB095)
Q4Dx6 indicates one dose every 4 days for a total of 6 doses

TABLE 27 — In vivo efficacy of humanized huAb13v1 anti-B7-H3 conjugates as monotherapy and in combination with docetaxel DAR/

	Conjugation	Dose	route/
ADC	Method	mg/kg/day	regimen	TGI max (%)	TGD (%)
EBC1
AB095	—	10	Q4Dx6/IP	0	0
huAb13v1-TX E2	2/I	10	Q4Dx6/IP	58	67
Docetaxel	—	7.5	QDx1/IV	85	80
huAb13v1-TX E2 +	2/I	10 + 7.5	Q4Dx6/IP + Q	140	140
Docetaxel			Dx1/IV
NCI-H1299
AB095		10	Q4Dx6/IP	0	0
huAb13v1-TX E2	2/I	10	Q4Dx6/IP	80	24
Docetaxel	—	7.5	QDx1/IV	87	48
huAb13v1-TX E2 +	2/I	10 + 7.5	Q4Dx6/IP + Q	97	83
Docetaxel			Dx1/IV
NCI-H1975
AB095		10	Q4Dx6/IP	0	0
huAb13v1-TX E2	2/I	10	Q4Dx6/IP	52	62
Docetaxel		7.5	QDx1/IV	81	77
huAb13v1-TX E2 +	2/I	10 + 7.5	Q4Dx6/IP + Q	92	108
Docetaxel			Dx1/IV
NCI-H1650
AB095	—	8	Q7Dx6/IP	0	0
huAb13v1-CZ	2/I	10	QDx1/IP	80	100
Docetaxel	—	7.5	QDx1/IV	84	143
huAb13v1-CZ + Docetaxel	—	10 + 7.5	QDx1/IP + Q	99	>600
			Dx1/IV
AB095 (a)	N.A.	10	IP/Q14Dx3	0	0
DTX	N.A.	7.5	IV/Q14Dx3	80*	158*
huAb13v1-WD E2	2/I	10	IP/Q14Dx3	67*	83*
huAb13v1-WD E2 +	2/I +	10 + 7.5	IP/Q14Dx3 +	98* ¥	>717* ¥
DTX)	N.A.		IV/Q14Dx3
huAb13v1-WD E2	2/.	3	IP/Q14Dx3	56*	75*
huAb13v1-WD E2 +	2/I +	3 + 7.5	IP/Q14Dx3 +	99* ¥	>717* ¥
DTX)	N.A.		IV/Q14Dx3
huAb13v1-WD E2	2/I	1	IP/Q14Dx3	60*	67*
huAb13v1-WD E2 +	2/I +	1 + 7.5	IP/Q14Dx3 +	88* ¥	467* ¥
DTX	N.A		IV/Q14Dx3
huAb13v1-AAA E2	2/I	10	IP/Q14Dx3	63*	117*
huAb13v1-AAA E2 +	2/I +	10 + 7.5	IP/Q14Dx3 +	99* ¥	>717* ¥
DTX	N.A		IV/Q14Dx3
huAb13v1-AAA E2	2/I	3	IP/Q14Dx3	60*	117*
huAb13v1-AAA E2 +	2/I +	3 + 7.5	IP/Q14Dx3 +	99* ¥	>717* ¥
DTX	N.A		IV/Q14Dx3
huAb13v1-AAA E2	2/I	1	IP/Q14Dx3	50*	67*
huAb13v1-AAA E2 +	2/I +	1 + 7.5	IP/Q14Dx3 +	92* ¥	>717* ¥
DTX	N.A		IV/Q14Dx3
(a) IgG1 mAb
*= p < 0.05 as compared to control treatment (AB095)
¥ = p < 0.05 as compared to the most active partner in a drug combination
N.A. = not applicable

SEQUENCE SUMMARY SEQ ID

NO:	Description	Amino Acid Sequence
1	chAb2 VH amino acid sequence	QVQLQQPGAELVKPGASVKLSCKA
		S GYTFTSYWMH WVKQRPGQGLEWI
		G MIHPDSGTTNYNEKFRS KATLTV
		DKSSSTAYMQLSSLTSEDSAVYYC
		AV YYGSTYWYFDV WGTGTTVTVSS
2	chAb2 VH CDR1 amino acid sequence
GYTFTSYWMH
3	chAb2 VH CDR2 amino acid sequence
MIHPDSGTTNYNEKFRS
4	chAb2 VH CDR3 amino acid sequence
YYGSTYWYFDV
5	chAb2 VL amino acid sequence	DVVMTQTPLSLPVSLGDQAYISC R
		SSQSLVHINGNTYLH WYRQKPGQS
		PKLLIY KVSNRFS GVPDRFSGSGS
		GIDFILKISRVEAEDLGVYFC SQS
		THFPFT FGSGTKLEIK
6	chAb2 VL CDR1 amino acid sequence
RSSQSLVHINGNTYLH
7	chAb2, chAb3, chAb10, huAb3VL.1,
KVSNRFS
	huAb3VL.1a, huAb3VL.1b, huAb3v2.1,
	huAb3v2.2, huAb3v2.3, huAb3v2.4,
	huAb3v2.5, huAb3v2.6, huAb3v2.7,
	huAb3v2.8, and huAb3v2.9 VL CDR2 amino
	acid sequence
8	chAb2 VL CDR3 amino acid sequence
SQSTHFPFT
9	chAb3 VH amino acid sequence	QVQLQQPGAELVKPGASVKLSCKA
		S GYTFSSYWMH WVKQRPGQGLEWI
		G LIHPDSGSTNYNEMFKN KATLTV
		DRSSSTAYVQLSSLTSEDSAVYFC
		AG GGRLYFDY WGQGTTLIVSS
10	chAb3, huAb3VH.1, huAb3VH.1a,
GYTFSSYWMH
	huAb3VH.1b, huAb3v2.1, huAb3v2.2,
	huAb3v2.3, huAb3v2.4, huAb3v2.5,
	huAb3v2.6, huAb3v2.7, huAb3v2.8, and
	huAb3v2.9 VH CDR1 amino acid sequence
11	chAb3, huAb3VH.1, huAb3VH.1a, and
LIHPDSGSTNYNEMFKN
	huAb3VH.1b VH CDR2 amino acid sequence
12	chAb3, huAb3VH.1, huAb3VH.1a,
GGRLYFDY
	huAb3VH.1b, huAb3v2.1, huAb3v2.2,
	huAb3v2.3, huAb3v2.4, huAb3v2.5,
	huAb3v2.6, huAb3v2.7, huAb3v2.8, and
	huAb3v2.9 VH CDR3 amino acid sequence
13	chAb3 VL amino acid sequence	DVVMTQTPLSLPVSLGDQASISC R
		SSQSLVHSNGDTYLR WYLQKPGQS
		PKLLIY KVSNRFS GVPDRFSGSGS
		GTDFTLKITRVEAEDLGVYFC SQS
		THVPYT FGGGTKLEIK
14	chAb3, huAb3VL.1, huAb3VL.1a, and
RSSQSLVHSNGDTYLR
	huAb3VL.1b VL CDR1 amino acid sequence
15	chAb3, huAb3VL.1, huAb3VL.1a,
SQSTHVPYT
	huAb3VL.1b, huAb3v2.1, huAb3v2.2,
	huAb3v2.3, huAb3v2.4, huAb3v2.5,
	huAb3v2.6, huAb3v2.7, huAb3v2.8, and
	huAb3v2.9 VL CDR3 amino acid sequence
16	chAb4 VH amino acid sequence	QVQLQQPGAELVKPGASVKLSCKA
		S GYSFTSYWMH WVKQRPGQGLEWI
		G MIHPNSGSNNYNEKFKS KATLTV
		DKSSNTAYMQLSSLTSEDSAVYYC
		AR RLGLHFDY WGQGTTLIVSS
17	chAb4 VH CDR1 amino acid sequence
GYSFTSYWMH
18	chAb4 VH CDR2 amino acid sequence
MIHPNSGSNNYNEKFKS
19	chAb4 VH CDR3 amino acid sequence
RLGLHFDY
20	chAb4 VL amino acid sequence	DIVMTQSQKFMSTPVGDRVSITC K
		ASQNVGTAVA WYQQKPGQSPKLLI
		Y SASNRYT GVPDRFTGSGSGTDFT
		LTISNMQSEDLADYFC QQYSSYPY
		T FGGGTKLEIK
21	chAb4 VL CDR1 amino acid sequence
KASQNVGTAVA
22	chAb4 VL CDR2 amino acid sequence	SASNRYT
23	chAb4 VL CDR3 amino acid sequence	QQYSSYPYT
24	chAb18 VH amino acid sequence	QVQLQQSAAELARPGASVKMSCKA
		S GYSFTSYTIH WVKQRPGQGLEWI
		G YINPNSRNTDYNQKFKD ETTLTA
		DRSSSTAYMQLISLTSEDSAVYYC
		AR YSGSTPYWYFDV WGAGTTVTVS
		S
25	chAb18, huAb18VH.1, huAb18VH.1a, and	GYSFTSYTIH
	huAb18VH.1b VH CDR1 amino acid sequence
26	chAb18, huAb18VH.1, and huAb18VH.1a VH	YINPNSRNTDYNQKFKD
	CDR2 amino acid sequence
27	chAb18, huAb18VH.1, huAb18VH.1a, and	YSGSTPYWYFDV
	huAb18VH.1b VH CDR3 amino acid sequence
28	chAb18 VL amino acid sequence	QIVLTQSPAILSASPGEKVTMTC R
		ASSSVSYMN WYQQKPGSSPKPWIY
		ATSNLAS GVPARFSVSVSGTSHSL
		TISRVEAEDAATYYC QQWSSNPLT
		FGAGTKLELK
29	chAb18, huAb18VL.1, huAb18VL.1a,	RASSSVSYMN
	huAb18VL.1b, huAb18VL.2, and
	huAb18VL.2a, VL CDR1 amino acid
	sequence
30	chAb18, huAb18VL.1, huAb18VL.1a,	ATSNLAS
	huAb18VL.1b, huAb18VL.2, and
	huAb18VL.2a, VL CDR2 amino acid
	sequence
31	chAb18, huAb18VL.1, huAb18VL.1a,	QQWSSNPLT
	huAb18VL.1b, huAb18VL.2, and
	huAb18VL.2a, VL CDR3 amino acid
	sequence
32	chAb13 VH amino acid sequence	DVQLQESGPDLVKPSQSLSLTCTV
		T GYSITSGYSWH WIRQFPGNKLEW
		MG YIHSSGSTNYNPSLKS RISINR
		DTSKNQFFLQLNSVTTEDTATYYC
		AG YDDYFEY WGQGTTLTVSS
33	chAb13, huAb13Vh.1, huAb13Vh.1a,	GYSITSGYSWH
	huAb13Vh.1b, huAb13v1, huAb13v2,
	huAb13v3, huAb13v4, huAb13v5, huAb13v6,
	huAb13v7, huAb13v8, and huAb13v9 VH
	CDR1 amino acid sequence
34	chAb13, huAb13Vh.1, huAb13Vh.1a,	YIHSSGSTNYNPSLKS
	huAb13Vh.1b, huAb13v1, huAb13v2,
	huAb13v3, huAb13v4, huAb13v5, huAb13v6,
	huAb13v7, huAb13v8, and huAb13v9 VH
	CDR2 amino acid sequence
35	chAb13, huAb13Vh.1, huAb13Vh.1a,	YDDYFEY
	huAb13Vh.1b, huAb13v1, huAb13v2,
	huAb13v3, huAb13v4, huAb13v5, huAb13v6,
	huAb13v7, huAb13v8, and huAb13v9 VH
	CDR3 amino acid sequence
36	chAb13 VL amino acid sequence	DIVMTQSQKFMSTSVGDRVSVTC K
		ASQNVGFNVA WYQQKPGQSPKALI
		Y SASYRYS GVPDRFTGSGSGTDFT
		LTISNVQSEDLAEYFC QQYNSYPF
		T FGSGTKLEIK
37	chAb13, huAb13VL.1, huAb13VL.1a,	KASQNVGFNVA
	huAb13VL.1b, huAb13v1, huAb13v2,
	huAb13v3, huAb13v4, huAb13v5, huAb13v6,
	huAb13v7, huAb13v8, and huAb13v9 VL
	CDR1 amino acid sequence
38	chAb13, huAb13VL.1, huAb13VL.1a,	SASYRYS
	huAb13VL.1b, huAb13v1, huAb13v2,
	huAb13v3, huAb13v4, huAb13v5, huAb13v6,
	huAb13v7, huAb13v8, and huAb13v9 VL
	CDR2 amino acid sequence
39	huAb13VL.1, huAb13VL.1a, huAb13VL.1b,	QQYNWYPFT
	huAb13v1, huAb13v2, huAb13v3, huAb13v4,
	huAb13v5, huAb13v6, huAb13v7, huAb13v8,
	and huAb13v9 VL CDR3 amino acid
	sequence
40	chAb12 VH amino acid sequence	EVQLVESGGGLVKPGGSLKLSCAA
		S GFTFSSYAMS WVRQTPEKRLEWV
		A TISSGTNYTYYPDSVKG RFTISR
		DNAKNTLYLQMTSLRSEDTAMYYC
		AR QGRYSWIAY WGQGTLVTVSA
41	chAb12 VH CDR1 amino acid sequence	GFTFSSYAMS
42	chAb12 VH CDR2 amino acid sequence	TISSGTNYTYYPDSVKG
43	chAb12 VH CDR3 amino acid sequence	QGRYSWIAY
44	chAb12 VL amino acid sequence	DIVLTQSPASLAVSLGQRATISC R
		ASKSVSTSDYSYMH WNQQKPGQPP
		KLLIY LASNLES GVPARFSGSGSG
		TDFTLNIHPVEEEDAATYYC QHSR
		ELLT FGAGTKLELK
45	chAb12 VL CDR1 amino acid sequence	RASKSVSTSDYSYMH
46	chAb12and chAb17 VL CDR2 amino acid	LASNLES
47	chAb12 VL CDR3 amino acid sequence	QHSRELLT
48	chAb14 VH amino acid sequence	EVKLVESGGGLVKPGGSLKLSCAA
		S GFTFSSYGMS WVRQTPEKRLEWV
		A TISGGGTNTYYPDSVEG RFTISR
		DNAKNFLYLQMSSLRSEDTALYYC
		AR HYGSQTMDY WGQGTSVTVSS
49	chAb14 and chAb8 VH CDR1 amino acid	GFTFSSYGMS
	sequence
50	chAb14 VH CDR2 amino acid sequence	TISGGGTNTYYPDSVEG
51	chAb14 VH CDR3 amino acid sequence	HYGSQTMDY
52	chAb14 VL amino acid sequence	DIQMTQSPASLSASVGETVTITC R
		TSGNIHNYLT WYQQKQGKSPQLLV
		Y NAKTLAD GVPSRFSGSGSGTQFS
		LKINSLQPEDFGSYYC QHFWSIMW
		T FGGGTKLEIK
53	chAb14 VL CDR1 amino acid sequence	RTSGNIHNYLT
54	chAb14 VL CDR2 amino acid sequence	NAKTLAD
55	chAb14 VL CDR3 amino acid sequence	QHFWSIMWT
56	chAb6 VH amino acid sequence	QVQLQQSGAELMKPGASVKISCKA
		T GYTFSRYWIE WVKQRPGHGLEWI
		G EILPGSGSTNYNEKFKG KATFTA
		DTSSNTAYMQVSSLTSEDSAVHYC
		AR RGYGYVPYALDY WGQGTSVIVS
		S
57	chAb6 VH CDR1 amino acid sequence	GYTFSRYWIE
58	chAb6 VH CDR2 amino acid sequence
EILPGSGSTNYNEKFKG
59	chAb6 VH CDR3 amino acid sequence	RGYGYVPYALDY
60	chAb6 VL amino acid sequence	EIQMTQTTSSLSASLGDRVTISC R
		ASQDISNSLN WYQQKPDGTVNLLI
		Y YTSRLYS GVPSRFSGSGSGTDYS
		LTISNLEQEDIATYFC QQGNTLPY
		T FGGGTKLEIK
61	chAb6 VL CDR1 amino acid sequence	RASQDISNSLN
62	chAb6 VL CDR2 amino acid sequence	YTSRLYS
63	chAb6 VL CDR3 amino acid sequence	QQGNTLPYT
64	chAb11 VH amino acid sequence	EVKLVESGGGLVQPGGSLRLSCAT
		S GFTFTNYYMS WVRQPPGKALEWL
		G FIRNKANDYTTEYSASVKG RFTI
		SRDNSQSILYLQMNTLRAEDSATY
		YCAR ESPGNPFAY WGQGTLVTVSA
65	chAb11 VH CDR1 amino acid sequence	GFTFTNYYMS
66	chAb11 VH CDR2 amino acid sequence	FIRNKANDYTTEYSASVKG
67	chAb11 VH CDR3 amino acid sequence	ESPGNPFAY
68	chAb11 VL amino acid sequence	DIVMTQSPSSLTVTAGEKVTMTC K
		SSQSLLNSGTQKNFLT WYQQKPGQ
		PPKLLIY WASTRES GVPDRFTGSG
		SGTDFTLTISSVQAEDLAVYFC QN
		DYIYPLT FGAGTKLELK
70	chAb11 VL CDR2 amino acid sequence	WASTRES
71	chAb11 VL CDR3 amino acid sequence	QNDYIYPLT
72	chAb16 VH amino acid sequence	EVKLLESGGGLVQPGGSLKLSCAA
		S GFDFSRYWMS WVRQAPGKGLEWI
		G EINPDSSTINYTPSLKD KFIISR
		DNAKNTLYLQMSKVRSEDTALYYC
		AR PGFGNYIYAMDY WGQGTSVTVS
		S
73	chAb16 VH CDR1 amino acid sequence	GFDFSRYWMS
74	chAb16 VH CDR2 amino acid sequence	EINPDSSTINYTPSLKD
75	chAb16 VH CDR3 amino acid sequence	PGFGNYIYAMDY
76	chAb16 VL amino acid sequence	DIQMTQTTSSLSASLGDRVTINC R
		ASQDISNFLN WYQQKPDGTVKLLI
		Y YTSRLYL GVPSRFSGSGSGTDYS
		LTISNLEQEDIATYFC QQGNTLPP
		T FGGGTKLEIK
77	chAb16 VL CDR1 amino acid sequence	RASQDISNFLN
78	chAb16 VL CDR2 amino acid sequence
YTSRLYL
79	chAb16 VL CDR3 amino acid sequence	QQGNTLPPT
80	chAb10 VH amino acid sequence	DVQLQESGPGLVKPSQSLSLTCTV
		T GYSITSDYAWN WIRQFPGNRLEW
		MG HINYSGITNYNPSLKS RISITR
		DTSKNQFFLQLYSVTTEDTATYFC
		AR RSLFYYYGSSLYAMDY WGQGTS
		VTVSS
81	chAb10 VH CDR1 amino acid sequence	GYSITSDYAWN
82	chAb10 VH CDR2 amino acid sequence	HINYSGITNYNPSLKS
83	chAb10 VH CDR3 amino acid sequence	RSLFYYYGSSLYAMDY
84	chAb10 VL amino acid sequence	DVVMTQSPFSLPVSLGDQASISC R
		SSQSLVHSNGNTYLH WYLQKPGQS
		PKLLIY KVSNRFS GVPDRFSGSGS
		GTDFTLKISRVEAEDLGVYFC SQS
		THVPWT FGGGTKLEIK
85	chAb10 VL CDR1 amino acid sequence	RSSQSLVHSNGNTYLH
86	chAb10 VL CDR3 amino acid sequence	SQSTHVPWT
87	chAb7 VH amino acid sequence	EVQLVESGENLVKPGGSLKLSCAA
		S GFSFRGYGMS WVRQTPDKRLEWV
		A AISTGGNYTYYPDSVQG RFTISR
		DNANNTLYLQMSSLKSEDTAMYYC
		AR RGGNYAGFAY WGQGTLVTVSA
88	chAb7 VH CDR1 amino acid sequence	GFSFRGYGMS
89	chAb7 VH CDR2 amino acid sequence	AISTGGNYTYYPDSVQG
90	chAb7 VH CDR3 amino acid sequence	RGGNYAGFAY
91	chAb7 VL amino acid sequence	DIQMTQSPASLSVSVGETVTITC R
		PSENIYSNLA WYQQKQGKSPQLLV
		Y AATNLAD GVPSRFSGSGSGTQYS
		LKINSLQSEDFGTYYC QHFWGTPF
		T FGSGTKLEIK
92	chAb7 VL CDR1 amino acid sequence	RPSENIYSNLA
93	chAb7 and chAb8 VL CDR2 amino acid	AATNLAD
94	chAb7 VL CDR3 amino acid sequence	QHFWGTPFT
95	chAb8 VH amino acid sequence	EVKLVESGGGLVKPGGSLKLSCAA
		S GFTFSSYGMS WVRQTPEKRLEWV
		A TISGGGNYTYCPDSVKG RFTISR
		DNAKNNLYLQMSSLRSEDTALYYC
		TR QRGYDYHYAMDF WGQGTSVIVS
		S
96	chAb8 VH CDR2 amino acid sequence	TISGGGNYTYCPDSVKG
97	chAb8 VH CDR3 amino acid sequence	QRGYDYHYAMDF
98	chAb8 VL amino acid sequence	DIQMTQSPASLSVSVGETVTITC R
		ASENIYSNLA WHQQKQGKSPQLLV
		Y AATNLAD GVPSRFSGNGSDTQYS
		LKINSLQSEDFGSYFC QNFWGTSW
		T FGGGTKLEIK
99	chAb8 VL CDR1 amino acid sequence	RASENIYSNLA
100	chAb8 VL CDR3 amino acid sequence	QNFWGTSWT
101	chAb17 VH amino acid sequence	EVKLVESGGGLVQPGGSLKLSCAA
		S GFTFSSYIMS WVRQTPEKRLEWV
		A SIVSSNITYYPDSMKG RFTISRD
		NARNILYLQMSSLKSEDTAMYYCA
		R SGTRAWFAY WGQGTLVTVSA
102	chAb17 VH CDR1 amino acid sequence	GFTFSSYIMS
103	chAb17 VH CDR2 amino acid sequence	SIVSSNITYYPDSMKG
104	chAb17 VH CDR3 amino acid sequence	SGTRAWFAY
105	chAb17 VL amino acid sequence	DIVLTQSPASLAVSLGQRATISC R
		ASKSVSTSAYSYMH WYQQKPGQPP
		KLLIY LASNLES GVPARFSGSGSG
		TDFTLNIHPVEEEDAATYYC QHSR
		ELPYT FGGGTKLEIK
106	chAb17 VL CDR1 amino acid sequence	RASKSVSTSAYSYMH
107	chAb17 VL CDR3 amino acid sequence	QHSRELPYT
108	chAb5 VH amino acid sequence	QVQLQQPGDELVKPGASVKLSCKT
		S GYTFTTDWMH WVKQRPGQGLEWI
		G MIHPNSGTTNYNEKFKS KAALTV
		DKSSSTACMQLSSLTSEDSAVYYC
		AR SYWKWYFDV WGIGTTVIVSS
109	chAb5 VH CDR1 amino acid sequence	GYTFTTDWMH
110	chAb5 VH CDR2 amino acid sequence	MIHPNSGTTNYNEKFKS
111	chAb5 VH CDR3 amino acid sequence	SYWKWYFDV
112	chAb5 VL amino acid sequence	QIVLTQSPAIMSASLGEEITLTC S
		ASSSVSYMH WYQQKSGTSPKLLIY
		STSNLAS GVPSRFSGSGSGTFYSL
		TISSVEAEDSADYYC HQWTSYMYT
		FGGGTKLEIK
113	chAb5 VL CDR1 amino acid sequence
SASSSVSYMH
114	chAb5 VL CDR2 amino acid sequence
STSNLAS
115	chAb5 VL CDR3 amino acid sequence
HQWTSYMYT
116	huAb18VH.1, huAbl8v1, and huAb18v5 VH	EVQLVQSGAEVKKPGSSVKVSCKA
	amino acid sequence	S GYSFTSYTIH WVRQAPGQGLEWM
		G YINPNSRNTDYNQKFKD RVTITA
		DKSTSTAYMELSSLRSEDTAVYYC
		AR YSGSTPYWYFDV WGQGTIVTVS
		S
117	huAb18VH.1a, huAb18v3, huAb18v8, and	EVQLVQSGAEVKKPGSSVKVSCKA
	huAb18v9 VH amino acid sequence	S GYSFTSYTIH WVRQAPGQGLEWI
		G YINPNSRNTDYNQKFKD RTTLTA
		DRSTSTAYMELSSLRSEDTAVYYC
		AR YSGSTPYWYFDV WGQGTIVTVS
		S
118	huAb18VH.1b, huAb18v2, huAb18v4,	EVQLVQSGAEVKKPGSSVKVSCKA
	huAb18v6, huAb18v7, and huAb18v10 VH	S GYSFTSYTIH WVRQAPGQGLEWM
	amino acid sequence	G YINPNSRNTDYAQKFQG RVTLTA
		DKSTSTAYMELSSLRSEDTAVYYC
		AR YSGSTPYWYFDV WGQGTIVTVS
		S
119	huAb18VH.1b VH CDR2 amino acid sequence
YINPNSRNTDYAQKFQG
120	huAb18VL.1, huAbl8v1, and huAb18v2 VL	DIQLTQSPSFLSASVGDRVTITC R
	amino acid sequence	ASSSVSYMN WYQQKPGKAPKLLIY
		ATSNLAS GVPSRFSGSGSGTEFTL
		TISSLQPEDFATYYC QQWSSNPLT
		FGQGTKLEIK
121	huAb18VL.1a, huAb18v3, and huAb18v4 VL	DIQLTQSPSFLSASVGDRVTITC R
	amino acid sequence	ASSSVSYMN WYQQKPGKSPKPWIY
		ATSNLAS GVPSRFSVSVSGTEHTL
		TISSLQPEDFATYYC QQWSSNPLT
		FGQGTKLEIK
122	huAb18VL.1b, huAb18v8, and huAb18v10 VL	DIQLTQSPSFLSASVGDRVTITC R
	amino acid sequence	ASSSVSYMN WYQQKPGKAPKPWIY
		ATSNLAS GVPSRFSVSGSGTEHTL
		TISSLQPEDFATYYC QQWSSNPLT
		FGQGTKLEIK
123	huAb18VL.2, huAb18v5, and huAb18v6 VL	EIVLTQSPDFQSVTPKEKVTITC R
	amino acid sequence	ASSSVSYMN WYQQKPDQSPKLLIK
		ATSNLAS GVPSRFSGSGSGTDFTL
		TINSLEAEDAATYYC QQWSSNPLT
		FGQGTKLEIK
124	huAb18VL.2a, huAb18v7, and huAb18v9 VL	EIVLTQSPDFQSVTPKEKVTITC R
	amino acid sequence	ASSSVSYMN WYQQKPDQSPKPWIY
		ATSNLAS GVPSRFSVSVSGTDHTL
		TINSLEAEDAATYYC QQWSSNPLT
		FGQGTKLEIK
125	huAb3VH.1, huAb3v1, and huAb3v4 VH	EVQLVQSGAEVKKPGSSVKVSCKA
	amino acid sequence	S GYTFSSYWMH WVRQAPGQGLEWM
		G LIHPDSGSTNYNEMFKN RVTITA
		DKSTSTAYMELSSLRSEDTAVYYC
		AR GGRLYFD YWGQGTTVIVSS
126	huAb3VH.1a, huAb3v3, and huAb3v6 VH	EVQLVQSGAEVKKPGSSVKVSCKA
	amino acid sequence	S GYTFSSYWMH WVRQAPGQGLEWI
		G LIHPDSGSTNYNEMFKN RATLTV
		DRSTSTAYVELSSLRSEDTAVYFC
		AG GGRLYFDY WGQGTTVIVSS
127	huAb3VH.1b, huAb3v2, and huAb3v5 VH	EVQLVQSGAEVKKPGSSVKVSCKA
	amino acid sequence	S GYTFSSYWMH WVRQAPGQGLEWI
		G LIHPDSGSTNYNEMFKN RATLTV
		DRSTSTAYMELSSLRSEDTAVYYC
		AG GGRLYFDY WGQGTTVIVSS
128	huAb3VL.1, huAb3v1, and huAb3v2 VL	DIVMTQSPLSLPVTPGEPASISC R
	amino acid sequence	SSQSLVHSNGDTYLR WYLQKPGQS
		PQLLIY KVSNRFS GVPDRFSGSGS
		GTDFTLKISRVEAEDVGVYYC SQS
		THVPYT FGGGTKVEIK
129	huAb3VL.1a and huAb3v3 VL amino acid	DVVMTQSPLSLPVTPGEPASISC R
	sequence	SSQSLVHSNGDTYLR WYLQKPGQS
		PQLLIY KVSNRFS GVPDRFSGSGS
		GTDFTLKISRVEAEDVGVYFC SQS
		THVPYT FGGGTKVEIK
130	huAb3VL.1b, huAb3v4, huAb3v5, and	DVVMTQSPLSLPVTPGEPASISC R
	huAb3v6 VL amino acid sequence	SSQSLVHSNGDTYLR WYLQKPGQS
		PQLLIY KVSNRFS GVPDRFSGSGS
		GTDFTLKISRVEAEDVGVYYC SQS
		THVPYT FGGGTKVEIK
131	huAb3v2.1, huAb3v2.2, and huAb3v2.3 VH	EVQLVQSGAEVKKPGSSVKVSCKA
	amino acid sequence	S GYTFSSYWMH WVRQAPGQGLEWI
		G LIHPWSGSTNYNEMFKN RATLTV
		DRSTSTAYMELSSLRSEDTAVYYC
		AG GGRLYFDY WGQGTTVIVSS
132	huAb3v2.1, huAb3v2.2, and huAb3v2.3 VH
LIHPWSGSTNYNEMFKN
	CDR2 amino acid sequence
133	huAb3v2.1, huAb3v2.4, and huAb3v2.7 VL	DIVMTQSPLSLPVTPGEPASISC R
	amino acid sequence	SSQSLVHSSGDTYLR WYLQKPGQS
		PQLLIY KVSNRFS GVPDRFSGSGS
		GTDFTLKISRVEAEDVGVYYC SQS
		THVPYT FGGGTKVEIK
134	huAb3v2.1, huAb3v2.4, and huAb3v2.7 VL
RSSQSLVHSSGDTYLR
	CDR1 amino acid sequence
135	huAb3v2.2, huAb3v2.5, and huAb3v2.8 VL	DIVMTQSPLSLPVTPGEPASISC R
	amino acid sequence	SSQSLVHSNRDTYLR WYLQKPGQS
		PQLLIY KVSNRFS GVPDRFSGSGS
		GTDFTLKISRVEAEDVGVYYC SQS
		THVPYT FGGGTKVEIK
136	huAb3v2.2, huAb3v2.5, and huAb3v2.8 VL
RSSQSLVHSNRDTYLR
	CDR1 amino acid sequence
137	huAb3v2.3, huAb3v2.6, and huAb3v2.9 VL	DIVMTQSPLSLPVTPGEPASISC R
	amino acid sequence	SSQSLVHSNQDTYLR WYLQKPGQS
		PQLLIY KVSNRFS GVPDRFSGSGS
		GTDFTLKISRVEAEDVGVYYC SQS
		THVPYT FGGGTKVEIK
138	huAb3v2.3, huAb3v2.6, and huAb3v2.9 VL
RSSQSLVHSNQDTYLR
	CDR1 amino acid sequence
139	huAb3v2.4, huAb3v2.5, and huAb3v2.6 VH	EVQLVQSGAEVKKPGSSVKVSCKA
	amino acid sequence	S GYTFSSYWMH WVRQAPGQGLEWI
		G LIHPESGSTNYNEMFKN RATLTV
		DRSTSTAYMELSSLRSEDTAVYYC
		AG GGRLYFDY WGQGTTVIVSS
140	huAb3v2.4, huAb3v2.5, and huAb3v2.6
LIHPESGSTNYNEMFKN
	VH CDR2 amino acid sequence
141	huAb3v2.7, huAb3v2.8, and huAb3v2.9 VH	EVQLVQSGAEVKKPGSSVKVSCKA
	amino acid sequence	S GYTFSSYWMH WVRQAPGQGLEWI
		G LIHPISGSTNYNEMFKN RATLTV
		DRSTSTAYMELSSLRSEDTAVYYC
		AG GGRLYFDY WGQGTTVIVSS
142	huAb3v2.7, huAb3v2.8, and huAb3v2.9 VH
LIHPISGSTNYNEMFKN
	CDR2 amino acid sequence
143	huAb13VL.1, huAb13v2, huAb13v5, and	DIQMTQSPSSLSASVGDRVTITC K
	huAb13v7 VL amino acid sequence	ASQNVGFNVA WYQQKPGKAPKLLI
		Y SASYRYS GVPSRFSGSGSGTDFT
		LTISSLQPEDFATYYC QQYNWYPF
		T FGQGTKLEIK
144	huAb13VL.1a, huAb13v1, huAb13v3, and	DIQMTQSPSSLSASVGDRVTITC K
	huAb13v8 VL amino acid sequence	ASQNVGFNVA WYQQKPGKSPKALI
		Y SASYRYS GVPSRFSGSGSGTDFT
		LTISSLQPEDFAEYFC QQYNWYPF
		T FGQGTKLEIK
145	huAB13VL.1b, huAb13v4, huAb13v6, and	DIQMTQSPSSLSASVGDRVTITC K
	huAb13v9 VL amino acid sequence	ASQNVGFNVA WYQQKPGKAPKLLI
		Y SASYRYS GVPSRFSGSGSGTDFT
		LTISSLQPEDFATYFC QQYNWYPF
		T FGQGTKLEIK
146	huAb13VH.1, huAb13v2, huAb13v3, and	EVQLQESGPGLVKPSETLSLTCAV
	huAb13v4 VH amino acid sequence	S GYSITSGYSWH WIRQPPGKGLEW
		IG YIHSSGSTNYNPSLKS RVTISV
		DTSKNQFSLKLSSVTAADTAVYYC
		AR YDDYFEY WGQGTTVTVSS
147	huAb13VH.1a, huAb13v1, huAb13v5, and	EVQLQESGPGLVKPSETLSLTCAV
	huAb13v6 VH amino acid sequence	T GYSITSGYSWH WIRQFPGNGLEW
		MG YIHSSGSTNYNPSLKS RISISR
		DTSKNQFFLKLSSVTAADTAVYYC
		AG YDDYFEY WGQGTTVTVSS
148	huAb13VH.1b, huAb13v7, huAb13v8, and	EVQLQESGPGLVKPSETLSLTCAV
	huAb13v9 VH amino acid sequence	S GYSITSGYSWH WIRQPPGNGLEW
		MG YIHSSGSTNYNPSLKS RITISR
		DTSKNQFSLKLSSVTAADTAVYYC
		AR YDDYFEY WGQGTTVTVSS
149	B7-H3 amino acid sequence (human)	MLRRRGSPGMGVHVGAALGALWFC
		LTGALEVQVPEDPVVALVGTDATL
		CCSFSPEPGFSLAQLNLIWQLTDT
		KQLVHSFAEGQDQGSAYANRTALF
		PDLLAQGNASLRLQRVRVADEGSF
		TCFVSIRDFGSAAVSLQVAAPYSK
		PSMTLEPNKDLRPGDTVTITCSSY
		QGYPEAEVFWQDGQGVPLIGNVIT
		SQMANEQGLFDVHSILRVVLGANG
		TYSCLVRNPVLQQDAHSSVTITPQ
		RSPTGAVEVQVPEDPVVALVGTDA
		TLRCSFSPEPGFSLAQLNLIWQLT
		DTKQLVHSFTEGRDQGSAYANRTA
		LFPDLLAQGNASLRLQRVRVADEG
		SFTCFVSIRDFGSAAVSLQVAAPY
		SKPSMTLEPNKDLRPGDTVTITCS
		SYRGYPEAEVFWQDGQGVPLTGNV
		TTSQMANEQGLFDVHSVLRVVLGA
		NGTYSCLVRNPVLQQDAHGSVTIT
		GQPMTFPPEALWVTVGLSVCLIAL
		LVALAFVCWRKIKQSCEEENAGAE
		DQDGEGEGSKTALQPLKHSDSKED
		DGQEIA
150	Human B7-H3-ECD (fc fusion)	MLRRRGSPGMGVHVGAALGALWFC
	Note: Fc sequence is underlined	LTGALEVQVPEDPVVALVGTDATL
		CCSFSPEPGFSLAQLNLIWQLTDT
		KQLVHSFAEGQDQGSAYANRTALF
		PDLLAQGNASLRLQRVRVADEGSF
		TCFVSIRDFGSAAVSLQVAAPYSK
		PSMTLEPNKDLRPGDTVTITCSSY
		QGYPEAEVFWQDGQGVPLTGNVTT
		SQMANEQGLFDVHSILRVVLGANG
		TYSCLVRNPVLQQDAHSSVTITPQ
		RSPTGAVEVQVPEDPVVALVGTDA
		TLRCSFSPEPGFSLAQLNLIWQLT
		DTKQLVHSFTEGRDQGSAYANRTA
		LFPDLLAQGNASLRLQRVRVADEG
		SFTCFVSIRDFGSAAVSLQVAAPY
		SKPSMTLEPNKDLRPGDTVTITCS
		SYRGYPEAEVFWQDGQGVPLTGNV
		TTSQMANEQGLFDVHSVLRVVLGA
		NGTYSCLVRNPVLQQDAHGSVTIT
		GQPMTFAAADK THTCPPCPAPEAE
GAPSVFLFPPKPKDTLMISRTPEV
TCVVVDVSHEDPEVKFNWYVDGVE
VHNAKTKPREEQYNSTYRVVSVLT
VLHQDWLNGKEYKCKVSNKALPAP
IEKTISKAKGQPREPQVYTLPPSR
EEMTKNQVSLTCLVKGFYPSDIAV
EWESNGQPENNYKTTPPVLDSDGS
FFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK
151	Mouse B7-H3-ECD (fc fusion)	MLRGWGGPSVGVCVRTALGVLCLC
	Note: Fc sequence is underlined	LTGAVEVQVSEDPVVALVDTDATL
		RCSFSPEPGFSLAQLNLIWQLTDT
		KQLVHSFTEGRDQGSAYSNRTALF
		PDLLVQGNASLRLQRVRVTDEGSY
		TCFVSIQDFDSAAVSLQVAAPYSK
		PSMTLEPNKDLRPGNMVTITCSSY
		QGYPEAEVFWKDGQGVPLTGNVTT
		SQMANERGLFDVHSVLRVVLGANG
		TYSCLVRNPVLQQDAHGSVTITGQ
		PLTFAAADK THTCPPCPAPEAEGA
PSVFLFPPKPKDTLMISRTPEVTC
VVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVL
HQDWLNGKEYKCKVSNKALPAPIE
KTISKAKGQPREPQVYTLPPSREE
MTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFF
LYSKLTVDKSRWQQGNVFSCSVMH
EALHNHYTQKSLSLSPGK
152	Human B7-H3-ECD (his tag)	MEFGLSWLFLVAILKGVQCGALEV
		QVPEDPVVALVGTDATLCCSFSPE
		PGFSLAQLNLIWQLTDTKQLVHSF
		AEGQDQGSAYANRTALFPDLLAQG
		NASLRLQRVRVADEGSFTCFVSIR
		DFGSAAVSLQVAAPYSKPSMTLEP
		NKDLRPGDTVTITCSSYQGYPEAE
		VFWQDGQGVPLTGNVTTSQMANEQ
		GLFDVHSILRVVLGANGTYSCLVR
		NPVLQQDAHSSVTITPQRSPTGAV
		EVQVPEDPVVALVGTDATLRCSFS
		PEPGFSLAQLNLIWQLTDTKQLVH
		SFTEGRDQGSAYANRTALFPDLLA
		QGNASLRLQRVRVADEGSFTCFVS
		IRDFGSAAVSLQVAAPYSKPSMTL
		EPNKDLRPGDTVTITCSSYRGYPE
		AEVFWQDGQGVPLTGNVTTSQMAN
		EQGLFDVHSVLRVVLGANGTYSCL
		VRNPVLQQDAHGSVTITGQPMTHH
		HHHH
153	Mouse B7-H3-ECD (his tag)	MEFGLSWLFLVAILKGVQCVEVQV
		SEDPVVALVDTDATLRCSFSPEPG
		FSLAQLNLIWQLTDTKQLVHSFTE
		GRDQGSAYSNRTALFPDLLVQGNA
		SLRLQRVRVTDEGSYTCFVSIQDF
		DSAAVSLQVAAPYSKPSMTLEPNK
		DLRPGNMVTITCSSYQGYPEAEVF
		WKDGQGVPLTGNVTTSQMANERGL
		FDVHSVLRVVLGANGTYSCLVRNP
		VLQQDAHGSVTITGQPLTFHHHHH
		H
154	Cynomolgus B7-H3-ECD (his tag)	MLHRRGSPGMGVHVGAALGALWFC
		LTGALEVQVPEDPVVALVGTDATL
		RCSFSPEPGFSLAQLNLIWQLTDT
		KQLVHSFTEGRDQGSAYANRTALF
		LDLLAQGNASLRLQRVRVADEGSF
		TCFVSIRDFGSAAVSLQVAAPYSK
		PSMTLEPNKDLRPGDTVTITCSSY
		RGYPEAEVFWQDGQGAPLTGNVTT
		SQMANEQGLFDVHSVLRVVLGANG
		TYSCLVRNPVLQQDAHGSITITPQ
		RSPTGAVEVQVPEDPVVALVGTDA
		TLRCSF
		SPEPGFSLAQLNLIWQLTDTKQLV
		HSFTEGRDQGSAYANRTALFLDLL
		AQGNASLRLQRVRVADEGSFTCFV
		SIRDFGSAAVSLQVAAPYSKPSMT
		LEPNKDLRPGDTVTITCSSYRGYP
		EAEVFWQDGQGAPLTGNVTTSQMA
		NEQGLFDVHSVLRVVLGANGTYSC
		LVRNPVLQQDAHGSVTITGQPMTF
		AAAHHHHHHHH
155	Amino acid sequence of IGHV1-69*06	QVQLVQSGAEVKKPGSSVKVSCKA
		SGGTFSSYAISWVRQAPGQGLEWM
		GGIIPIFGTANYAQKFQGRVTITA
		DKSTSTAYMELSSLRSEDTAVYYC
		AR
156	Amino acid sequence of IGHJ6*01	WGQGTTVTVSS
157	Amino acid sequence of IGKV1-9*01	DIQLTQSPSFLSASVGDRVTITCR
		ASQGISSYLAWYQQKPGKAPKLLI
		YAASTLQSGVPSRFSGSGSGTEFT
		LTISSLQPEDFATYYCQQLNSYPP
158	Amino acid sequence of IGKJ2*01	FGQGTKLEIK
159	Ig gamma-1 constant region	ASTKGPSVFPLAPSSKSTSGGTAA
		LGCLVKDYFPEPVTVSWNSGALTS
		GVHTFPAVLQSSGLYSLSSVVTVP
		SSSLGTQTYICNVNHKPSNTKVDK
		KVEPKSCDKTHTCPPCPAPELLGG
		PSVFLFPPKPKDTLMISRTPEVTC
		VVVDVSHEDPEVKFNWYVDGVEVH
		NAKTKPREEQYNSTYRVVSVLTVL
		HQDWLNGKEYKCKVSNKALPAPIE
		KTISKAKGQPREPQVYTLPPSREE
		MTKNQVSLTCLVKGFYPSDIAVEW
		ESNGQPENNYKTTPPVLDSDGSFF
		LYSKLTVDKSRWQQGNVFSCSVMH
		EALHNHYTQKSLSLSPGK
160	Ig gamma-1 constant region mutant	ASTKGPSVFPLAPSSKSTSGGTAA
		LGCLVKDYFPEPVTVSWNSGALTS
		GVHTFPAVLQSSGLYSLSSVVTVP
		SSSLGTQTYICNVNHKPSNTKVDK
		KVEPKSCDKTHTCPPCPAPEAAGG
		PSVFLFPPKPKDTLMISRTPEVTC
		VVVDVSHEDPEVKFNWYVDGVEVH
		NAKTKPREEQYNSTYRVVSVLTVL
		HQDWLNGKEYKCKVSNKALPAPIE
		KTISKAKGQPREPQVYTLPPSREE
		MTKNQVSLTCLVKGFYPSDIAVEW
		ESNGQPENNYKTTPPVLDSDGSFF
		LYSKLTVDKSRWQQGNVFSCSVMH
		EALHNHYTQKSLSLSPGK
161	Ig Kappa constant region	RTVAAPSVFIFPPSDEQLKSGTAS
		VVCLLNNFYPREAKVQWKVDNALQ
		SGNSQESVTEQDSKDSTYSLSSTL
		TLSKADYEKHKVYACEVTHQGLSS
		PVTKSFNRGEC
162	Ig Lambda constant region	QPKAAPSVTLFPPSSEELQANKAT
		LVCLISDFYPGAVTVAWKADSSPV
		KAGVETTTPSKQSNNKYAASSYLS
		LTPEQWKSHRSYSCQVTHEGSTVE
		KTVAPTECS
163	Amino acid sequence of IGKV6-21*01	EIVLTQSPDFQSVTPKEKVTITCR
		ASQSIGSSLHWYQQKPDQSPKLLI
		KYASQSFSGVPSRFSGSGSGTDFT
		LTINSLEAEDAATYYCHQSSSLPX
164	Amino acid sequence of IGKV2-28*01	DIVMTQSPLSLPVTPGEPASISCR
		SSQSLLHSNGYNYLDWYLQKPGQS
		PQLLIYLGSNRASGVPDRFSGSGS
		GTDFTLKISRVEAEDVGVYYCMQA
		LQTPP
165	Amino acid sequence of IGKJ4*01	FGGGTKVEIK
166	Amino acid sequence of IGHV-b*01(0-1)	QVQLQESGPGLVKPSETLSLTCAV
		SGYSISSGYYWGWIRQPPGKGLEW
		IGSIYHSGSTYYNPSLKSRVTISV
		DTSKNQFSLKLSSVTAADTAVYYC
		AR
167	Amino acid sequence of IGKv1-39*01	DIQMTQSPSSLSASVGDRVTITCR
		ASQSISSYLNWYQQKPGKAPKLLI
		YAASSLQSGVPSRFSGSGSGTDFT
		LTISSLQPEDFATYYCQQSYSTPP
168	Amino acid sequence of huAb13v1 heavy	EVQLQESGPGLVKPSETLSLTCAV
	chain	TGYSITSGYSWHWIRQFPGNGLEW
	Note: Ig gamma-1 constant region mutant	MGYIHSSGSTNYNPSLKSRISISR
	sequence is underlined.	DTSKNQFFLKLSSVTAADTAVYYC
		AGYDDYFEYWGQGTTVTVSS ASTK
GPSVFPLAPSSKSTSGGTAALGCL
VKDYFPEPVTVSWNSGALTSGVHT
FPAVLQSSGLYSLSSVVTVPSSSL
GTQTYICNVNHKPSNTKVDKKVEP
KSCDKTHTCPPCPAPEAAGGPSVF
LFPPKPKDTLMISRTPEVICVVVD
VSHEDPEVKFNWYVDGVEVHNAKT
KPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTIS
KAKGQPREPQVYTLPPSREEMTKN
QVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSK
LTVDKSRWQQGNVFSCSVMHEALH
NHYTQKSLSLSPGK
169	Amino acid sequence of huAb13v1 light	DIQMTQSPSSLSASVGDRVTITCK
	chain	ASQNVGFNVAWYQQKPGKSPKALI
	Note: Ig kappa constant region sequence	YSASYRYSGVPSRFSGSGSGTDFT
	is underlined.	LTISSLQPEDFAEYFCQQYNWYPF
		TFGQGTKLEIK RTVAAPSVFIFPP
SDEQLKSGTASVVCLLNNFYPREA
KVQWKVDNALQSGNSQESVTEQDS
KDSTYSLSSTLTLSKADYEKHKVY
ACEVTHQGLSSPVTKSFNRGEC
170	Amino acid sequence of huAb3v2.5 heavy	EVQLVQSGAEVKKPGSSVKVSCKA
	chain	SGYTFSSYWMHWVRQAPGQGLEWI
	Note: Ig gamma-1 constant region mutant	GLIHPESGSTNYNEMFKNRATLTV
	sequence is underlined.	DRSTSTAYMELSSLRSEDTAVYYC
		AGGGRLYFDYWGQGTTVTVSS AST
KGPSVFPLAPSSKSTSGGTAALGC
LVKDYFPEPVTVSWNSGALTSGVH
TFPAVLQSSGLYSLSSVVTVPSSS
LGTQTYICNVNHKPSNTKVDKKVE
PKSCDKTHTCPPCPAPEAAGGPSV
FLFPPKPKDTLMISRTPEVTCVVV
DVSHEDPEVKFNWYVDGVEVHNAK
TKPREEQYNSTYRVVSVLTVLHQD
WLNGKEYKCKVSNKALPAPIEKTI
SKAKGQPREPQVYTLPPSREEMTK
NQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYS
KLTVDKSRWQQGNVFSCSVMHEAL
HNHYTQKSLSLSPGK
171	Amino acid sequence of huAb3v2.5 light	DIVMTQSPLSLPVTPGEPASISCR
	chain	SSQSLVHSNRDTYLRWYLQKPGQS
	Note: Ig kappa constant region sequence	PQLLIYKVSNRFSGVPDRFSGSGS
	is underlined.	GTDFTLKISRVEAEDVGVYYCSQS
		THVPYTFGGGTKVEIK RTVAAPSV
FIFPPSDEQLKSGTASVVCLLNNF
YPREAKVQWKVDNALQSGNSQESV
TEQDSKDSTYSLSSTLTLSKADYE
KHKVYACEVTHQGLSSPVTKSFNR
GEC
172	Amino acid sequence of huAb3v2.6 heavy	EVQLVQSGAEVKKPGSSVKVSCKA
	chain	SGYTFSSYWMHWVRQAPGQGLEWI
	Note: Ig gamma-1 constant region mutant	GLIHPESGSTNYNEMFKNRATLTV
	sequence is underlined.	DRSTSTAYM
		ELSSLRSEDTAVYYCAGGGRLYFD
		YWGQGTTVTVSS ASTKGPSVFPLA
PSSKSTSGGTAALGCLVKDYFPEP
VTVSWNSGALTSGVHTFPAVLQSS
GLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHT
CPPCPAPEAAGGPSVFLFPPKPKD
TLMISRTPEVTCVVVDVSHEDPEV
KFNWYVDGVEVHNAKTKPREEQYN
STYRVVSVLTVLHQDWLNGKEYKC
KVSNKALPAPIEKTISKAKGQPRE
PQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKT
TPPVLDSDGSFFLYSKLTVDKSRW
QQGNVFSCSVMHEALHNHYTQKSL
SLSPGK
173	Amino acid sequence of huAb3v2.6 light	DIVMTQSPLSLPVTPGEPASISCR
	chain	SSQSLVHSNQDTYLRWYLQKPGQS
	Note: Ig kappa constant region sequence	PQLLIYKVSNRFSGVPDRFSGSGS
	is underlined.	GTDFTLKISRVEAEDVGVYYCSQS
		THVPYTFGGGTKVEIK RTVAAPSV
FIFPPSDEQLKSGTASVVCLLNNF
YPREAKVQWKVDNALQSGNSQESV
TEQDSKDSTYSLSSTLTLSKADYE
KHKVYACEVTHQGLSSPVTKSFNR
GEC
174	Amino acid sequence of	QVQLVQSGAEVKKPGSSVKVSCKA
	IGHV1-69*06 IGHJ6	SGGTFSSYAISWVRQAPGQGLEWM
		GGIIPIFGTANYAQKFQGRVTITA
		DKSTSTAYMELSSLRSEDTAVYYC
		ARXXXXXXXXWGQGTTVTVSS
175	Amino acid sequence of	DIVMTQSPLSLPVTPGEPASISCR
	IGKV2-28*01 IGKJ4	SSQSLLHSNGYNYLDWYLQKPGQS
		PQLLIYLGSNRASGVPDRFSGSGS
		GTDFTLKISRVEAEDVGVYYCXXX
		XXXXXXFGGGTKVEIK
176	Amino acid sequence of IGHV4-b IGHJ6	QVQLQESGPGLVKPSETLSLTCAV
		SGYSISSGYYWGWIRQPPGKGLEW
		IGSIYHSGSTYYNPSLKSRVTISV
		DTSKNQFSLKLSSVTAADTAVYYC
		ARXXXXXXXWGQGTTVTVSS
177	Amino acid sequence of IGKV1-39 IGKJ2	DIQMTQSPSSLSASVGDRVTITCR
		ASQSISSYLNWYQQKPGKAPKLLI
		YAASSLQSGVPSRFSGSGSGTDFT
		LTISSLQPEDFATYYCXXXXXXXX
		XFGQGTKLEIK
178	Amino acid sequence of huAb3 VL1	DIVMTQSPLSLPVTPGEPASISCR
	variants	SSQSLVHSXGDTYLRWYLQKPGQS
	Note: X can be any amino acid except:	PQLLIYKVSNRFSGVPDRFSGSGS
	M, C, N, D, or Q	GTDFTLKISRVEAEDVGVYYCSQS
		THVPYTFGGGTKVEIK
179	Amino acid sequence of huAb3 VL1	DIVMTQSPLSLPVTPGEPASISCR
	variants	SSQSLVHSNXDTYLRWYLQKPGQS
	Note: X can be any amino acid except:	PQLLIYKVSNRFSGVPDRFSGSGS
	M, C, G, S, N, or P	GTDFTLKISRVEAEDVGVYYCSQS
		THVPYTFGGGTKVEIK
180	Amino acid sequence of huAb3 VH1b	EVQLVQSGAEVKKPGSSVKVSCKA
	variants	SGYTFSSYWMHWVRQAPGQGLEWI
	Note: X can be any amino acid except:	GLIHPXSGSTNYNEMFKNRATLTV
	M, C, N, D, or Q	DRSTSTAYMELSSLRSEDTAVYYC
		AGGGRLYFDYWGQGTTVTVSS
181	Amino acid sequence of huAb3 VH1b	EVQLVQSGAEVKKPGSSVKVSCKA
	variants	SGYTFSSYWMHWVRQAPGQGLEWI
	Note: X can be any amino acid except:	GLIHPDXGSTNYNEMFKNRATLTV
	M, C, G, S, N, or P	DRSTSTAYMELSSLRSEDTAVYYC
		AGGGRLYFDYWGQGTTVTVSS
182	chAb13 VL CDR3 amino acid sequence	QQYNSYPFT

description truncated at 500,000 characters

Stored text is truncated at the source; the tail of the description is not held.

Claims as granted

3 claims

Classifications

7 codes

IPC · International Patent Classification

Section A — Human necessities

A61K47/68
A61K31/437
A61K31/519
A61K39/00
A61K31/498

Section C — Chemistry; metallurgy

C07K16/00
C07K16/28

Claim changes

Soon

Coming soonHow the claims changed between publication and grant

See which claims were amended, added or cancelled during examination, with every added and removed word marked.

AmendedAddedCancelledUnchanged

The published claims of this application are not paired with the granted ones in what we hold.

File wrapper

⤢ drag to zoom

USPTOApplicanthover for detail · click to open

Pendency

2.9 y

1,063 days filing → grant

Office actions

after a restriction

Responses

1 RCE

Examiner

Maher M Haddad

art unit 1644 · TC 1600

Law firm

Law firm · Log in to unlock

Citations: 590 back · 2 forward

See the full prosecution history — every USPTO and applicant action on this file, in order.

Documents

Log in to open the documents of this file: the application as filed, every office action and response, the notice of allowance.

Chain of title

⤢ drag to zoom

Titlehover for detail · click to open

See the full assignment history — every owner this patent has passed through, with recordation dates and reel/frame numbers.