Lipid formulations for delivery of messenger RNA

This application is a continuation application of U.S. application Ser. No. 15/451,312, filed Mar. 6, 2017; which is a divisional application of U.S. application Ser. No. 14/521,161, filed Oct. 22, 2014; which claims priority to U.S. Provisional Application Ser. No. 61/894,299, filed Oct. 22, 2013 and U.S. Provisional Application Ser. No. 61/953,516, filed Mar. 14, 2014, the disclosures of which are hereby incorporated by reference.

The present specification makes reference to a Sequence Listing (submitted electronically as .txt file named “MRT-1121US3_MIT17255_SL.txt” on Jul. 3, 2018). The .txt file was generated on Jul. 3, 2018 and is 32,504 bytes in size. The entire contents of the Sequence Listing are herein incorporated by reference.

Delivery of nucleic acids has been explored extensively as a potential therapeutic option for certain disease states. In particular, RNA interference (RNAi) has been the subject of significant research and clinical development. While RNAi, such as short interfering RNA (siRNA), may have therapeutic potential, it is of little use in treating diseases involving deficiency of one or more proteins. messenger RNA (mRNA) therapy has become an increasingly important option for treatment of various diseases, in particular, for those associated with deficiency of one or more proteins.

The present invention provides improved methods and compositions for highly efficient delivery and expression of mRNA and encoded protein in vivo. The invention is based, in part, on the surprising discovery that liposomes based on a particular class of cationic lipids, such as, those having a structure of formula I-c described herein, are unexpectedly effective in delivering mRNA and producing encoded protein in vivo, more effective even as compared to those cationic lipids that were considered to be among the best in delivering mRNA in the prior art. Indeed, prior to the present invention, cationic lipids have been extensively explored as an important component of liposomes typically used to encapsulate nucleic acids including mRNA for in vivo delivery. Due to the uniquely fragile and long structure of mRNA and the complicated in vivo translation process, cationic lipids used in the liposomes typically play two roles. First, cationic lipids promote interaction with negatively charged mRNA during encapsulation, circulation and endocytosis, thereby capturing and protecting the mRNA. Then, once inside cytosol, cationic lipids need to be able to release the mRNA so that the mRNA can be translated to produce encoded protein. Some cationic lipids, in particular, those known as titratable cationic lipids are particularly effective in delivering mRNA. One example of such cationic lipids known to be capable of efficient delivery of mRNA is C12-200. Surprisingly, the present inventors found that cationic lipids described herein can be even more effective in delivering various mRNA in vivo, than those best known in the prior art including C12-200. For example, as shown in the Examples below, liposome particles incorporating a cationic lipid described herein (e.g., cKK-E12) resulted in close to 50% higher protein expression of human Factor IX protein detected in the plasma of administered mice, as compared to C12-200-based liposome particles. Furthermore, the plasma residence time of different proteins expressed from mRNA delivered by cKK-E12 based liposomes is sustained up to 7 days or longer post a single administration. Thus, the present inventors have demonstrated that this class of cationic lipids having a structure of formula I-c described herein (e.g., cKK-E12) can be uniquely useful in delivering mRNA for highly efficient and sustained production of protein (e.g., therapeutic protein) in vivo. The present invention therefore permits an improved mRNA therapy that can significantly reduce required amount of mRNA and associated lipids, administration frequency, and possible side effects, providing more potent, safer, and patient friendly mRNA therapy for various diseases.

In one aspect, the present invention provides methods of delivering messenger RNA (mRNA) in vivo, including administering to a subject in need of delivery a composition comprising an mRNA encoding a protein, encapsulated within a liposome such that the administering of the composition results in the expression of the protein encoded by the mRNA in vivo, wherein the liposome comprises a cationic lipid of formula I-c:

or a pharmaceutically acceptable salt thereof,

wherein:

p is an integer of between 1 and 9, inclusive; each instance of R 2 is independently hydrogen or optionally substituted C 1-6 alkyl; each instance of R 6 and R 7 is independently a group of the formula (i), (ii), or (iii); Formulae (i), (ii), and (iii) are:

wherein:

each instance of R′ is independently hydrogen or optionally substituted alkyl; X is O, S, or NR X , wherein R X is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, or a nitrogen protecting group; Y is O, S, or NR Y , wherein R Y is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, or a nitrogen protecting group; R P is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, an oxygen protecting group when attached to an oxygen atom, a sulfur protecting group when attached to a sulfur atom, or a nitrogen protecting group when attached to a nitrogen atom; and R L is optionally substituted C 1-50 alkyl, optionally substituted C 2-50 alkenyl, optionally substituted C 2-50 alkynyl, optionally substituted heteroC 1-50 alkyl, optionally substituted heteroC 2-50 alkenyl, optionally substituted heteroC 2-50 alkynyl, or a polymer.

In another aspect, the present invention provides methods of treating a disease or disorder including administering to subject in need of treatment a composition comprising an mRNA encoding a therapeutic protein encapsulated within a liposome such that the administering of the composition results in the expression of the protein encoded by the mRNA in one or more tissues affected by the disease or disorder, wherein the liposome comprises a cationic lipid having a structure of formula I-c.

In another aspect, the present invention provides compositions for delivery of messenger RNA (mRNA) comprising an mRNA encoding a protein encapsulated within a liposome, wherein the liposome comprises a cationic lipid having a structure of formula I-c.

In some embodiments, a suitable cationic lipid is cKK-E12:

In some embodiments, a suitable liposome further comprises one or more non-cationic lipids, one or more cholesterol-based lipids and/or one or more PEG-modified lipids. In some embodiments, the one or more non-cationic lipids are selected from distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoyl-phosphatidylethanolamine (POPE), dioleoyl-phosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl-phosphatidyl-ethanolamine (DSPE), 16-O-monomethyl PE, 16-O-dimethyl PE, 18-1-trans PE, 1-stearoyl-2-oleoyl-phosphatidyethanolamine (SOPE), or a mixture thereof.

In some embodiments, a suitable liposome further comprises one or more cholesterol-based lipids. In some embodiments, the one or more cholesterol-based lipids are selected from cholesterol, PEGylated cholesterol and DC-Chol (N,N-dimethyl-N-ethylcarboxamidocholesterol), 1,4-bis(3-N-oleylamino-propyl)piperazine.

In some embodiments, a suitable liposome further comprises one or more PEG-modified lipids. In some embodiments, the one or more PEG-modified lipids comprise a poly(ethylene) glycol chain of up to 5 kDa in length covalently attached to a lipid with alkyl chain(s) of C 6 -C 20 length. In some embodiments, a PEG-modified lipid is a derivatized ceramide such as N-Octanoyl-Sphingosine-1-[Succinyl(Methoxy Polyethylene Glycol)-2000]. In some embodiments, a PEG-modified or PEGylated lipid is PEGylated cholesterol or Dimyristoylglycerol (DMG)-PEG-2K.

In some embodiments, a suitable liposome comprises cKK-E12, DOPE, cholesterol and DMG-PEG2K.

In some embodiments, the cationic lipid (e.g., cKK-E12) constitutes about 30-50% (e.g., about 30-45%, about 30-40%, about 35-50%, about 35-45%, or about 35-40%) of the liposome by molar ratio. In some embodiments, the cationic lipid (e.g., cKK-E12) constitutes about 30%, about 35%, about 40%, about 45%, or about 50% of the liposome by molar ratio.

In particular embodiments, the ratio of cKK-E12:DOPE:cholesterol:DMG-PEG2K is approximately 40:30:20:10 by molar ratio. In particular embodiments, the ratio of cKK-E12:DOPE:cholesterol:DMG-PEG2K is approximately 40:30:25:5 by molar ratio. In particular embodiments, the ratio of cKK-E12:DOPE:cholesterol:DMG-PEG2K is approximately 40:32:25:3 by molar ratio.

In some embodiments, a suitable liposome has a size of or less than about 500 nm, 450 nm, 400 nm, 350 nm, 300 nm, 250 nm, 200 nm, 150 nm, 125 nm, 110 nm, 100 nm, 95 nm, 90 nm, 85 nm, 80 nm, 75 nm, 70 nm, 65 nm, 60 nm, 55 nm, or 50 nm.

In some embodiments, a composition according to the invention is administered intravenously. In some embodiments, a composition according to the invention is administered via pulmonary delivery. In some embodiments, the pulmonary delivery is by aerosolization, inhalation, nebulization or instillation. In some embodiments, a composition according to the invention is administered intrathecally. In some embodiments, the composition is formulated as respirable particles, nebulizable lipid, or inhalable dry powder.

In some embodiments, the expression of the protein encoded by the mRNA is detectable in liver, kidney, heart, spleen, serum, brain, skeletal muscle, lymph nodes, skin, and/or cerebrospinal fluid.

In some embodiments, the expression of the protein encoded by the mRNA is detectable 6, 12, 18, 24, 30, 36, 42, 48, 54, 60, 66, and/or 72 hours after the administration. In some embodiments, the expression of the protein encoded by the mRNA is detectable 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, and/or 7 days after the administration. In some embodiments, the expression of the protein encoded by the mRNA is detectable 1 week, 2 weeks, 3 weeks, and/or 4 weeks after the administration. In some embodiments, the expression of the protein encoded by the mRNA is detectable after a month after the administration.

In some embodiments, the protein encoded by the mRNA is a cytosolic protein. In some embodiments, the protein encoded by the mRNA is a secreted protein. In some embodiments, the protein encoded by the mRNA is an enzyme. In some embodiments, the mRNA has a length of or greater than about 0.5 kb, 1 kb, 1.5 kb, 2 kb, 2.5 kb, 3 kb, 3.5 kb, 4 kb, 4.5 kb, or 5 kb. In some embodiments, the protein encoded by the mRNA is Argininosuccinate Synthetase (ASS1), Factor IX, survival of motor neuron 1, or phenylalanine hydroxylase.

In some embodiments, the mRNA is administered at a dose ranging from about 0.1-5.0 mg/kg body weight, for example about 0.1-4.5, 0.1-4.0, 0.1-3.5, 0.1-3.0, 0.1-2.5, 0.1-2.0, 0.1-1.5, 0.1-1.0, 0.1-0.5, 0.1-0.3, 0.3-5.0, 0.3-4.5, 0.3-4.0, 0.3-3.5, 0.3-3.0, 0.3-2.5, 0.3-2.0, 0.3-1.5, 0.3-1.0, 0.3-0.5, 0.5-5.0, 0.5-4.5, 0.5-4.0, 0.5-3.5, 0.5-3.0, 0.5-2.5, 0.5-2.0, 0.5-1.5, or 0.5-1.0 mg/kg body weight. In some embodiments, the mRNA is administered at a dose of or less than about 5.0, 4.5, 4.0, 3.5, 3.0, 2.5, 2.0, 1.5, 1.0, 0.8, 0.6, 0.5, 0.4, 0.3, 0.2, or 0.1 mg/kg body weight.

In some embodiments, the mRNA comprises one or more modified nucleotides. In some embodiments, the one or more modified nucleotides comprise pseudouridine, N-1-methyl-pseudouridine, 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyl adenosine, 5-methylcytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5-propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, O(6)-methylguanine, and/or 2-thiocytidine. In some embodiments, the mRNA is unmodified.

Other features, objects, and advantages of the present invention are apparent in the detailed description, drawings and claims that follow. It should be understood, however, that the detailed description, the drawings, and the claims, while indicating embodiments of the present invention, are given by way of illustration only, not limitation. Various changes and modifications within the scope of the invention will become apparent to those skilled in the art.

The drawings are for illustration purposes only not for limitation.

FIG. 1 shows an exemplary graph of the levels of human factor IX (FIX) detected in the serum of treated mice 24 hours after administration of C12-200 or cKK-E12 liposomes containing FIX mRNA.

FIG. 2 shows an exemplary graph of FIX detected in the plasma of mice treated with 0.1, 0.3, 0.6, 1.0, or 3.0 mg/kg of one of two ratios of FIX mRNA containing cKK-E12 liposomes either 6 or 24 hours after administration.

FIG. 3 shows an exemplary graph of the level of ASS1 protein detected in the livers of mice treated with 0.1, 0.3, 0.6, 1.0, or 3.0 mg/kg of ASS1 mRNA-containing cKK-E12 liposomes 24 hours after administration.

FIG. 4 shows exemplary western blot analyses of ASS1 protein levels in the liver 24 hours post administration of 0.1, 0.3, 0.6, 1.0, or 3.0 mg/kg of cKK-E12 liposomes containing ASS1 mRNA.

FIG. 5 shows an exemplary graph of ASS1 protein levels in the liver of mice 0.5, 3, 6, 12, 24, 48, 72 hours after a single IV injection of ASS1 mRNA containing cKK-E12 liposomes (1 mg/kg). Also shown is the level of ASS1 protein 7 days after administration.

FIG. 6 shows exemplary western blot analyses of ASS1 protein levels in the liver 0.5, 3, 6, 12, 24, 48, 72 hours after a single IV injection of 1 mg/kg ASS1 mRNA containing cKK-E12 liposomes. Also shown is the level of ASS1 protein 7 days after administration.

FIG. 7 —shows detection of human ASS1 messenger RNA via in situ hybridization in the livers of treated mice. Exogenous mRNA is observable for at least 72 hr post-administration after a single dose (1.0 mg/kg) of ASS1 mRNA-loaded MD1-based lipid nanoparticles. Human ASS1 mRNA is detectable in sinusoidal cells as well as hepatocytes.

FIG. 8 —shows exemplary immunohistochemical staining of ASS1 protein levels in mouse liver 24 hours after administration of 1 mg/kg ASS1 mRNA containing cKK-E12 lipid nanoparticles. Human ASS1 protein is detectable in sinusoidal cells as well as hepatocytes.

FIG. 9 shows low magnification (4×) immunohistochemical staining of ASS1 protein levels in mouse liver 24 hours after administration of 1 mg/kg ASS1 mRNA containing cKK-E12 liposomes. A comparison to untreated mouse liver (left) demonstrates the widespread distribution of human ASS1 protein throughout the liver.

FIG. 10 shows exemplary results illustrating that cKK-E12 lipid nanoparticles efficiently delivered FL mRNA via nebulization. Mice were exposed to milligram of encapsulated FL mRNA and analysis was performed 24 hours post-exposure.

FIG. 11 illustrates detection via western blot of human SMN-1 protein derived from exogenous hSMN-1 mRNA that was transfected into BHK-21 cells. Various antibodies specific to human SMN were employed: (A) anti-SMN 4F11 antibody at 1:1,000 dilution; (B) Pierce PA5-27309 a-SMN antibody at 1:10,000 dilution; and (C) LSBio C138149 a-SMN antibody at 1:10,000 dilution.

FIG. 12A-C illustrates multiplex nucleic acid in situ detection of human Survival of Motor Neuron (hSMN-1) mRNA in (A) Cervical, (B) Thoracic and (C) Lumbar spinal tissue, 24 hours post intrathecal delivery.

FIG. 13 illustrates positive detection of human SMN-1 protein produced in the spinal cord of a rat 24 hours post-intrathecal administration of human SMN-1 mRNA-loaded lipid nanoparticles. Anti-human SMN 4F11 antibody was employed at 1:2500 dilution. Panel A represents treated rat spinal cord tissue and panel B represents untreated rat spinal cord tissue.

FIG. 14 In vivo transfection of CFTR knockout mice with C-terminal His 10 tagged (SEQ ID NO: 11) codon-optimized human CFTR mRNA encapsulated within either a lipid (cKK-E12) or polymeric (PEI) nanoparticle formulation. Following nebulized delivery of each respective mRNA formulation, Right and Left lung tissue lysate was collected and analyzed for CFTR expression by Western blot using anti-His antibody 1187. Control CFTR knockout lung tissue and CFTR-His 10 HEK293 lysate (“His 10 ” disclosed as SEQ ID NO: 11) was used as a negative and positive controls respectively.

FIG. 15 illustrates positive detection of active firefly luciferase (FFL) protein in a treated pig lung via luminescence upon exposure to FFL/CO-CFTR-C-His 10 mRNA (“His 10 ” disclosed as SEQ ID NO: 11) encapsulated cKK-E12 lipid nanoparticles. Pigs were treated with 1 mg FFL+9 mg CO-CFTR-C-His 10 mRNA (“His 10 ” disclosed as SEQ ID NO: 11) encapsulated lipid nanoparticles via nebulization using a Pari jet nebulizer and sacrificed 24 hours post-treatment. FFL luminescence was visualized using an IVIS bioluminometer.

In order for the present invention to be more readily understood, certain terms are first defined below. Additional definitions for the following terms and other terms are set forth throughout the specification. The publications and other reference materials referenced herein to describe the background of the invention and to provide additional detail regarding its practice are hereby incorporated by reference.

Amino acid: As used herein, term “amino acid,” in its broadest sense, refers to any compound and/or substance that can be incorporated into a polypeptide chain. In some embodiments, an amino acid has the general structure H E N—C(H)(R)—COHO. In some embodiments, an amino acid is a naturally occurring amino acid. In some embodiments, an amino acid is a synthetic amino acid; in some embodiments, an amino acid is a d-amino acid; in some embodiments, an amino acid is an 1-amino acid. “Standard amino acid” refers to any of the twenty standard 1-amino acids commonly found in naturally occurring peptides. “Nonstandard amino acid” refers to any amino acid, other than the standard amino acids, regardless of whether it is prepared synthetically or obtained from a natural source. As used herein, “synthetic amino acid” encompasses chemically modified amino acids, including but not limited to salts, amino acid derivatives (such as amides), and/or substitutions. Amino acids, including carboxyl- and/or amino-terminal amino acids in peptides, can be modified by methylation, amidation, acetylation, protecting groups, and/or substitution with other chemical groups that can change the peptide's circulating half-life without adversely affecting their activity. Amino acids may participate in a disulfide bond. Amino acids may comprise one or posttranslational modifications, such as association with one or more chemical entities (e.g., methyl groups, acetate groups, acetyl groups, phosphate groups, formyl moieties, isoprenoid groups, sulfate groups, polyethylene glycol moieties, lipid moieties, carbohydrate moieties, biotin moieties, etc.). The term “amino acid” is used interchangeably with “amino acid residue,” and may refer to a free amino acid and/or to an amino acid residue of a peptide. It will be apparent from the context in which the term is used whether it refers to a free amino acid or a residue of a peptide.

Animal: As used herein, the term “animal” refers to any member of the animal kingdom. In some embodiments, “animal” refers to humans, at any stage of development. In some embodiments, “animal” refers to non-human animals, at any stage of development. In certain embodiments, the non-human animal is a mammal (e.g., a rodent, a mouse, a rat, a rabbit, a monkey, a dog, a cat, a sheep, cattle, a primate, and/or a pig). In some embodiments, animals include, but are not limited to, mammals, birds, reptiles, amphibians, fish, insects, and/or worms. In some embodiments, an animal may be a transgenic animal, genetically-engineered animal, and/or a clone.

Approximately or about: As used herein, the term “approximately” or “about,” as applied to one or more values of interest, refers to a value that is similar to a stated reference value. In certain embodiments, the term “approximately” or “about” refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value).

Delivery: As used herein, the term “delivery” encompasses both local and systemic delivery. For example, delivery of mRNA encompasses situations in which an mRNA is delivered to a target tissue and the encoded protein is expressed and retained within the target tissue (also referred to as “local distribution” or “local delivery”), and situations in which an mRNA is delivered to a target tissue and the encoded protein is expressed and secreted into patient's circulation system (e.g., serum) and systematically distributed and taken up by other tissues (also referred to as “systemic distribution” or “systemic delivery).

Expression: As used herein, “expression” of a nucleic acid sequence refers to translation of an mRNA into a polypeptide, assemble multiple polypeptides (e.g., heavy chain or light chain of antibody) into an intact protein (e.g., antibody) and/or post-translational modification of a polypeptide or fully assembled protein (e.g., antibody). In this application, the terms “expression” and “production,” and grammatical equivalent, are used inter-changeably.

Functional: As used herein, a “functional” biological molecule is a biological molecule in a form in which it exhibits a property and/or activity by which it is characterized.

Half-life: As used herein, the term “half-life” is the time required for a quantity such as nucleic acid or protein concentration or activity to fall to half of its value as measured at the beginning of a time period.

Improve, increase, or reduce: As used herein, the terms “improve,” “increase” or “reduce,” or grammatical equivalents, indicate values that are relative to a baseline measurement, such as a measurement in the same individual prior to initiation of the treatment described herein, or a measurement in a control subject (or multiple control subject) in the absence of the treatment described herein. A “control subject” is a subject afflicted with the same form of disease as the subject being treated, who is about the same age as the subject being treated.

In Vitro: As used herein, the term “in vitro” refers to events that occur in an artificial environment, e.g., in a test tube or reaction vessel, in cell culture, etc., rather than within a multi-cellular organism.

In Vivo: As used herein, the term “in vivo” refers to events that occur within a multi-cellular organism, such as a human and a non-human animal. In the context of cell-based systems, the term may be used to refer to events that occur within a living cell (as opposed to, for example, in vitro systems).

Isolated: As used herein, the term “isolated” refers to a substance and/or entity that has been (1) separated from at least some of the components with which it was associated when initially produced (whether in nature and/or in an experimental setting), and/or (2) produced, prepared, and/or manufactured by the hand of man. Isolated substances and/or entities may be separated from about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% of the other components with which they were initially associated. In some embodiments, isolated agents are about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% pure. As used herein, a substance is “pure” if it is substantially free of other components. As used herein, calculation of percent purity of isolated substances and/or entities should not include excipients (e.g., buffer, solvent, water, etc.).

Local distribution or delivery: As used herein, the terms “local distribution,” “local delivery,” or grammatical equivalent, refer to tissue specific delivery or distribution. Typically, local distribution or delivery requires a protein (e.g., enzyme) encoded by mRNAs be translated and expressed intracellularly or with limited secretion that avoids entering the patient's circulation system.

messenger RNA (mRNA): As used herein, the term “messenger RNA (mRNA)” refers to a polynucleotide that encodes at least one polypeptide. mRNA as used herein encompasses both modified and unmodified RNA. mRNA may contain one or more coding and non-coding regions. mRNA can be purified from natural sources, produced using recombinant expression systems and optionally purified, chemically synthesized, etc. Where appropriate, e.g., in the case of chemically synthesized molecules, mRNA can comprise nucleoside analogs such as analogs having chemically modified bases or sugars, backbone modifications, etc. An mRNA sequence is presented in the 5′ to 3′ direction unless otherwise indicated. In some embodiments, an mRNA is or comprises natural nucleosides (e.g., adenosine, guanosine, cytidine, uridine); nucleoside analogs (e.g., 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyl adenosine, 5-methylcytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5-propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, O(6)-methylguanine, and 2-thiocytidine); chemically modified bases; biologically modified bases (e.g., methylated bases); intercalated bases; modified sugars (e.g., 2′-fluororibose, ribose, 2′-deoxyribose, arabinose, and hexose); and/or modified phosphate groups (e.g., phosphorothioates and 5′-N-phosphoramidite linkages).

In some embodiments, the mRNA comprises one or more nonstandard nucleotide residues. The nonstandard nucleotide residues may include, e.g., 5-methyl-cytidine (“5mC”), pseudouridine (“ψU”), and/or 2-thio-uridine (“2sU”). See, e.g., U.S. Pat. No. 8,278,036 or WO2011012316 for a discussion of such residues and their incorporation into mRNA. The mRNA may be RNA, which is defined as RNA in which 25% of U residues are 2-thio-uridine and 25% of C residues are 5-methylcytidine. Teachings for the use of RNA are disclosed US Patent Publication US20120195936 and international publication WO2011012316, both of which are hereby incorporated by reference in their entirety. The presence of nonstandard nucleotide residues may render an mRNA more stable and/or less immunogenic than a control mRNA with the same sequence but containing only standard residues. In further embodiments, the mRNA may comprise one or more nonstandard nucleotide residues chosen from isocytosine, pseudoisocytosine, 5-bromouracil, 5-propynyluracil, 6-aminopurine, 2-aminopurine, inosine, diaminopurine and 2-chloro-6-aminopurine cytosine, as well as combinations of these modifications and other nucleobase modifications. Certain embodiments may further include additional modifications to the furanose ring or nucleobase. Additional modifications may include, for example, sugar modifications or substitutions (e.g., one or more of a 2′-O-alkyl modification, a locked nucleic acid (LNA)). In some embodiments, the RNAs may be complexed or hybridized with additional polynucleotides and/or peptide polynucleotides (PNA). In embodiments where the sugar modification is a 2′-O-alkyl modification, such modification may include, but are not limited to a 2′-deoxy-2′-fluoro modification, a 2′-O-methyl modification, a 2′-O-methoxyethyl modification and a 2′-deoxy modification. In certain embodiments, any of these modifications may be present in 0-100% of the nucleotides—for example, more than 0%, 1%, 10%, 25%, 50%, 75%, 85%, 90%, 95%, or 100% of the constituent nucleotides individually or in combination.

Nucleic acid: As used herein, the term “nucleic acid,” in its broadest sense, refers to any compound and/or substance that is or can be incorporated into a polynucleotide chain. In some embodiments, a nucleic acid is a compound and/or substance that is or can be incorporated into a polynucleotide chain via a phosphodiester linkage. In some embodiments, “nucleic acid” refers to individual nucleic acid residues (e.g., nucleotides and/or nucleosides). In some embodiments, “nucleic acid” refers to a polynucleotide chain comprising individual nucleic acid residues. In some embodiments, “nucleic acid” encompasses RNA as well as single and/or double-stranded DNA and/or cDNA.

Patient: As used herein, the term “patient” or “subject” refers to any organism to which a provided composition may be administered, e.g., for experimental, diagnostic, prophylactic, cosmetic, and/or therapeutic purposes. Typical patients include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and/or humans). In some embodiments, a patient is a human. A human includes pre and post natal forms.

Pharmaceutically acceptable: The term “pharmaceutically acceptable” as used herein, refers to substances that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

Systemic distribution or delivery: As used herein, the terms “systemic distribution,” “systemic delivery,” or grammatical equivalent, refer to a delivery or distribution mechanism or approach that affect the entire body or an entire organism. Typically, systemic distribution or delivery is accomplished via body's circulation system, e.g., blood stream. Compared to the definition of “local distribution or delivery.”

Subject: As used herein, the term “subject” refers to a human or any non-human animal (e.g., mouse, rat, rabbit, dog, cat, cattle, swine, sheep, horse or primate). A human includes pre- and post-natal forms. In many embodiments, a subject is a human being. A subject can be a patient, which refers to a human presenting to a medical provider for diagnosis or treatment of a disease. The term “subject” is used herein interchangeably with “individual” or “patient.” A subject can be afflicted with or is susceptible to a disease or disorder but may or may not display symptoms of the disease or disorder.

Substantially: As used herein, the term “substantially” refers to the qualitative condition of exhibiting total or near-total extent or degree of a characteristic or property of interest. One of ordinary skill in the biological arts will understand that biological and chemical phenomena rarely, if ever, go to completion and/or proceed to completeness or achieve or avoid an absolute result. The term “substantially” is therefore used herein to capture the potential lack of completeness inherent in many biological and chemical phenomena.

Target tissues: As used herein, the term “target tissues” refers to any tissue that is affected by a disease to be treated. In some embodiments, target tissues include those tissues that display disease-associated pathology, symptom, or feature.

Therapeutically effective amount: As used herein, the term “therapeutically effective amount” of a therapeutic agent means an amount that is sufficient, when administered to a subject suffering from or susceptible to a disease, disorder, and/or condition, to treat, diagnose, prevent, and/or delay the onset of the symptom(s) of the disease, disorder, and/or condition. It will be appreciated by those of ordinary skill in the art that a therapeutically effective amount is typically administered via a dosing regimen comprising at least one unit dose.

Treating: As used herein, the term “treat,” “treatment,” or “treating” refers to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of and/or reduce incidence of one or more symptoms or features of a particular disease, disorder, and/or condition. Treatment may be administered to a subject who does not exhibit signs of a disease and/or exhibits only early signs of the disease for the purpose of decreasing the risk of developing pathology associated with the disease.

The present invention provides, among other things, methods and compositions for delivering mRNA in vivo using improved liposomes incorporating cationic lipids described herein.

Liposomes for mRNA Delivery

As used herein, the term “liposome” refers to any lamellar, multilamellar, or solid lipid nanoparticle vesicle. Typically, a liposome as used herein can be formed by mixing one or more lipids or by mixing one or more lipids and polymer(s). Thus, the term “liposome” as used herein encompasses both lipid and polymer based nanoparticles. In particular, a liposome according to the present invention incorporates a cationic lipid described herein. As a non-limiting example, a cationic lipid suitable for the present invention is cKK-E12, or (3,6-bis(4-(bis(2-hydroxydodecyl)amino)butyl)piperazine-2,5-dione), as described in more detail below. A suitable liposome may also contain second or additional cationic lipids, helper lipids (e.g., non-cationic lipids and/or cholesterol-based lipids), PEG-modified lipids, and/or polymers.

In some embodiments, cationic lipid(s) (e.g., cKK-E12) constitute(s) about 30-50% (e.g., about 30-45%, about 30-40%, about 35-50%, about 35-45%, or about 35-40%) of the liposome by molar ratio. In some embodiments, the cationic lipid (e.g., cKK-E12) constitutes about 30%, about 35%, about 40%, about 45%, or about 50% of the liposome by molar ratio.

Cationic Lipids

In some embodiments, provided liposomes or compositions provided comprise a cationic lipid according to formula I:

or a pharmaceutically acceptable salt thereof,

wherein:

In some embodiments, a cationic lipid in a provided composition or method is a compound of formula I. In some embodiments, a cationic lipid in a provided composition or method is a compound of formula I, wherein the compound comprises one or more basic groups. In some embodiments, a cationic lipid in a provided composition or method is a compound of formula I, wherein the compound comprises one or more amino groups.

In certain embodiments, a group of formula (i) represents a group of formula (i-a) or a group of formula (i-b):

wherein each variable is independently as defined above and described herein. In some embodiments, a group of formula (i) is a group of formula (i-a). In some embodiments, a group of formula (i) is a group of formula (i-b).

In some embodiments, at least one instance of R 1 is a group of formula (iv). In some embodiments, at least one instance of R 1 is a group of formula (iv), wherein at least one of R 6 and R 7 is a group of formula (i), (ii) or (iii). In some embodiments, at least one instance of R 1 is a group of formula (iv), wherein each of R 6 and R 7 is independently a group of formula (i), (ii) or (iii).

In some embodiments, each R 1 is independently a group of formula (iv). In some embodiments, each R 1 is independently a group of formula (iv), wherein at least one of R 6 and R 7 is a group of formula (i), (ii) or (iii). In some embodiments, each R 1 is independently a group of formula (iv), wherein each of R 6 and R 7 is independently a group of formula (i), (ii) or (iii). In some embodiments, each R 1 is independently a group of formula (iv), wherein each of R 6 and R 7 is independently a group of formula (i). In some embodiments, each R 1 is independently a group of formula (iv), wherein each of R 6 and R 7 is independently a group of formula (ii). In some embodiments, each R 1 is independently a group of formula (iv), wherein each of R 6 and R 7 is independently a group of formula (iii). In some embodiments, each R 1 is independently a group of formula (iv), wherein each of R 6 and R 7 is independently a group of formula (i-a). In some embodiments, each R 1 is independently a group of formula (iv), wherein each of R 6 and R 7 is independently a group of formula (i-b).

In some embodiments, each instance of R′ is hydrogen.

In some embodiments, L is an optionally substituted alkylene.

In some embodiments, a group of formula (iv) is of formula

wherein q is an integer between 1 and 50, inclusive, and each of R 6 and R 7 is independently as defined above and described herein.

As generally defined above, p is an integer of between 1 and 9, inclusive. In certain embodiments, p is 1. In certain embodiments, p is 2. In certain embodiments, p is 3. In certain embodiments, p is 4. In certain embodiments, p is 5. In certain embodiments, p is 6. In certain embodiments, p is 7. In certain embodiments, p is 8. In certain embodiments, p is 9.

In some embodiments, p is 1. In some embodiments, a compound of formula I is a compound of formula (I-a):

or a pharmaceutically acceptable salt thereof, wherein each variable is independently as defined above and described herein.

In some embodiments, p is 2. In some embodiments, a compound of formula I is a compound of formula (I-p2):

or a pharmaceutically acceptable salt thereof, wherein each variable is independently as defined above and described herein.

In some embodiments, p is 3. In some embodiments, a compound of formula I is a compound of formula (I-p3):

or a pharmaceutically acceptable salt thereof, wherein each variable is independently as defined above and described herein.

In some embodiments, p is 4. In some embodiments, a compound of formula I is a compound of formula (I-p4):

or a pharmaceutically acceptable salt thereof, wherein each variable is independently as defined above and described herein.

In some embodiments, p is 5. In some embodiments, a compound of formula I is a compound of formula (I-p5):

or a pharmaceutically acceptable salt thereof, wherein each variable is independently as defined above and described herein.

In some embodiments, p is 6. In some embodiments, a compound of formula I is a compound of formula (I-p6):

or a pharmaceutically acceptable salt thereof, wherein each variable is independently as defined above and described herein.

In some embodiments, p is 7. In some embodiments, a compound of formula I is a compound of formula (I-p7):

or a pharmaceutically acceptable salt thereof, wherein each variable is independently as defined above and described herein.

In some embodiments, p is 8. In some embodiments, a compound of formula I is a compound of formula (I-p8):

or a pharmaceutically acceptable salt thereof, wherein each variable is independently as defined above and described herein.

In some embodiments, p is 9. In some embodiments, a compound of formula I is a compound of formula (I-p9):

or a pharmaceutically acceptable salt thereof, wherein each variable is independently as defined above and described herein.

As generally defined above, each instance of Q is independently O, S, or NR Q , wherein R Q is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, a nitrogen protecting group, or a group of the formula (i), (ii), or (iii).

In certain embodiments, at least one instance of Q is O. In certain embodiments, each instance of Q is O. In certain embodiments, at least one instance of Q is S. In certain embodiments, each instance of Q is S. In certain embodiments, at least one instance of Q is NR Q , wherein R Q is as defined above and described herein. In certain embodiments, each instance of Q is NR Q , wherein each R Q is independently as defined above and described herein. In certain embodiments, each instance of R Q is independently hydrogen or a group of the formula (i), (ii), or (iii).

As generally defined above, R Q is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, a nitrogen protecting group, or a group of the formula (i), (ii) or (iii).

In some embodiments, R Q is hydrogen. In some embodiments, R Q is optionally substituted alkyl. In some embodiments, R Q is optionally substituted alkenyl. In some embodiments, R Q is optionally substituted alkynyl. In some embodiments, R Q is carbocyclyl. In some embodiments, R Q is optionally substituted heterocyclyl. In some embodiments, R Q is optionally substituted aryl. In some embodiments, R Q is optionally substituted heteroaryl. In some embodiments, R Q is a nitrogen protecting group. In some embodiments, R Q is a group of formula (i), (ii) or (iii). In some embodiments, R Q is a group of formula (i). In some embodiments, R Q is a group of formula (ii). In some embodiments, R Q is a group of formula (iii).

As generally defined above, each instance of R 1 is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, halogen, —OR A1 , —N(R A1 ) 2 , or —SR A1 , or a group of formula (iv), wherein each of R A1 and formula (iv) is independently as defined above and described herein.

In some embodiments, R 1 is hydrogen.

In certain embodiments, R 1 is optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl. In certain embodiments, at least one instance of R 1 is optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl.

In certain embodiments, R 1 is optionally substituted alkyl; e.g., optionally substituted C 1-6 alkyl, optionally substituted C 2-6 alkyl, optionally substituted C 3-6 alkyl, optionally substituted C 4-6 alkyl, optionally substituted C 4-5 alkyl, or optionally substituted C 3-4 alkyl. In certain embodiments, at least one instance of R 1 is optionally substituted alkyl; e.g., optionally substituted C 1-6 alkyl, optionally substituted C 2-6 alkyl, optionally substituted C 3-6 alkyl, optionally substituted C 4-6 alkyl, optionally substituted C 4-5 alkyl, or optionally substituted C 3-4 alkyl.

In certain embodiments, R 1 is optionally substituted alkenyl, e.g., optionally substituted C 2-6 alkenyl, optionally substituted C 3-6 alkenyl, optionally substituted C 4-6 alkenyl, optionally substituted C 4-5 alkenyl, or optionally substituted C 3-4 alkenyl. In certain embodiments, at least one instance of R 1 is optionally substituted alkenyl, e.g., optionally substituted C 2-6 alkenyl, optionally substituted C 3-6 alkenyl, optionally substituted C 4-6 alkenyl, optionally substituted C 4-5 alkenyl, or optionally substituted C 3-4 alkenyl.

In certain embodiments, R 1 is optionally substituted alkynyl, e.g., optionally substituted C 2-6 alkynyl, optionally substituted C 3-6 alkynyl, optionally substituted C 4-6 alkynyl, optionally substituted C 4-5 alkynyl, or optionally substituted C 3-4 alkynyl. In certain embodiments, at least one instance of R 1 is optionally substituted alkynyl, e.g., optionally substituted C 2-6 alkynyl, optionally substituted C 3-6 alkynyl, optionally substituted C 4-6 alkynyl, optionally substituted C 4-5 alkynyl, or optionally substituted C 3-4 alkynyl.

In certain embodiments, R 1 is optionally substituted carbocyclyl, e.g., optionally substituted C 3-10 carbocyclyl, optionally substituted C 5-8 carbocyclyl, optionally substituted C 5-6 carbocyclyl, optionally substituted C 5 carbocyclyl, or optionally substituted C 6 carbocyclyl. In certain embodiments, at least one instance of R 1 is optionally substituted carbocyclyl, e.g., optionally substituted C 3-10 carbocyclyl, optionally substituted C 5-8 carbocyclyl, optionally substituted C 5-6 carbocyclyl, optionally substituted C 5 carbocyclyl, or optionally substituted C 6 carbocyclyl.

In some embodiments, R 1 is optionally substituted heterocyclyl, e.g., optionally substituted 3-14 membered heterocyclyl, optionally substituted 3-10 membered heterocyclyl, optionally substituted 5-8 membered heterocyclyl, optionally substituted 5-6 membered heterocyclyl, optionally substituted 5-membered heterocyclyl, or optionally substituted 6-membered heterocyclyl. In certain embodiments, at least one instance of R 1 is optionally substituted heterocyclyl, e.g., optionally substituted 3-14 membered heterocyclyl, optionally substituted 3-10 membered heterocyclyl, optionally substituted 5-8 membered heterocyclyl, optionally substituted 5-6 membered heterocyclyl, optionally substituted 5-membered heterocyclyl, or optionally substituted 6-membered heterocyclyl.

In some embodiments, R 1 is optionally substituted aryl. In some embodiments, R 1 is optionally substituted phenyl. In some embodiments, R 1 is phenyl. In some embodiments, R 1 is substituted phenyl. In certain embodiments, at least one instance of R 1 is optionally substituted aryl, e.g., optionally substituted phenyl.

In some embodiments, R 1 is optionally substituted heteroaryl, e.g., optionally substituted 5-14 membered heteroaryl, optionally substituted 5-10 membered heteroaryl, optionally substituted 5-6 membered heteroaryl, optionally substituted 5 membered heteroaryl, or optionally substituted 6 membered heteroaryl. In certain embodiments, at least one instance of R 1 is optionally substituted heteroaryl, e.g., optionally substituted 5-14 membered heteroaryl, optionally substituted 5-10 membered heteroaryl, optionally substituted 5-6 membered heteroaryl, optionally substituted 5 membered heteroaryl, or optionally substituted 6 membered heteroaryl.

In some embodiments, R 1 is halogen. In some embodiments, R 1 is —F. In some embodiments, R 1 is —Cl. In some embodiments, R 1 is —Br. In some embodiments, R 1 is —I.

In some embodiments, R 1 is —OR A1 , wherein R A1 is as defined above and described herein. In some embodiments, R 1 is —N(R A1 ) 2 , wherein each R A1 is independently as defined above and described herein. In some embodiments, R 1 is —SR A1 , wherein R A1 is as defined above and described herein.

In some embodiments, an R 1 alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl group may be substituted. In some embodiments, an R 1 alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl group may be substituted with an optionally substituted amino group. In some embodiments, an R 1 alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl group may be substituted with an optionally substituted hydroxyl group. In some embodiments, an R 1 alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl group may be substituted with an optionally substituted thiol group. In any of the above embodiments, an R 1 alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl group may be substituted, for example, with an optionally substituted amino group (e.g., —NR 6 R), an optionally substituted hydroxyl group (e.g., —OR 6 ), an optionally substituted thiol group (e.g., —SR 6 ), or with a group of formula (i), (ii), or (iii), wherein each instance of R 6 and R 7 is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, a nitrogen protecting group when attached to a nitrogen atom, an oxygen protecting group when attached to an oxygen atom, and a sulfur protecting group when attached to a sulfur atom, or a group of formula (i), (ii), or (iii).

In some embodiments, R 1 is an optionally substituted natural amino acid side chain. In some embodiments, R 1 is a natural amino acid side chain. In some embodiments, R 1 is an optionally substituted unnatural amino acid side chain. In some embodiments, R 1 is an unnatural amino acid side chain.

In certain embodiments, each instance of R 1 is the same. In certain embodiments, at least one R 1 group is different. In certain embodiments, each R 1 group is different.

In certain embodiments, R 1 is an alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl group substituted with an amino group of the formula —NR 6 R 7 .

In certain embodiments, R 1 is a group of formula (iv):

wherein:

L is an optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted heteroalkylene, optionally substituted heteroalkenylene, optionally substituted heteroalkynylene, optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, or optionally substituted heteroarylene, or combination thereof, and

each of R 6 and R 7 is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, a nitrogen protecting group, or a group of formula (i), (ii) or (iii):

wherein each of R′, Y, R P , R L and X is independently as defined above and described herein.

In some embodiments, at least one instance of R 1 is an alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl group substituted with an amino group of the formula —NR 6 R 7 . In some embodiments, at least one instance of R 1 is a group of formula (iv). In some embodiments, at least one instance of R 1 is a group of formula (iv), wherein at least one instance of R 6 and R 7 is a group of the formula (i), (ii) or (iii). In some embodiments, at least one instance of R 1 is a group of formula (iv), wherein each instance of R 6 and R 7 is a group of the formula (i), (ii) or (iii). In some embodiments, at least one instance of R 1 is a group of formula (iv), wherein each instance of R 6 and R 7 is a group of the formula (i). In some embodiments, at least one instance of R 1 is a group of formula (iv), wherein each instance of R 6 and R 7 is a group of the formula (ii). In some embodiments, at least one instance of R 1 is a group of formula (iv), wherein each instance of R 6 and R 7 is a group of the formula (iii).

In some embodiments, each instance of R 1 is a group of formula (iv). In some embodiments, each instance of R 1 is a group of formula (iv), wherein each instance of R 6 and R 7 is a group of the formula (i), (ii) or (iii). In some embodiments, each instance of R 1 is a group of formula (iv), wherein each instance of R 6 and R 7 is a group of the formula (i), (ii) or (iii). In some embodiments, each instance of R 1 is a group of formula (iv), wherein each instance of R 6 and R 7 is a group of the formula (i). In some embodiments, each instance of R 1 is a group of formula (iv), wherein each instance of R 6 and R 7 is a group of the formula (ii). In some embodiments, each instance of R 1 is a group of formula (iv), wherein each instance of R 6 and R 7 is a group of the formula (iii).

In certain embodiments, at least two instances of R 1 is a group of formula (iv). In certain embodiments, at least three instances of R 1 is a group of formula (iv). In certain embodiments, at least four instances of R 1 is a group of formula (iv). In certain embodiments, at least five instances of R 1 is a group of formula (iv). In certain embodiments, at least six instances of R 1 is a group of formula (iv). In certain embodiments, at least seven instances of R 1 is a group of formula (iv). In certain embodiments, at least eight instances of R 1 is a group of formula (iv). In certain embodiments, at least nine instances of R 1 is a group of formula (iv). In certain embodiments, each instance of R 1 is a group of formula (iv).

In certain embodiments, L is an optionally substituted alkylene; e.g., optionally substituted C 1-50 alkylene, optionally substituted C 1-40 alkylene, optionally substituted C 1-30 alkylene, optionally substituted C 1-20 alkylene, optionally substituted C 4-20 alkylene, optionally substituted C 6-20 alkylene, optionally substituted C 8-20 alkylene, optionally substituted C 10-20 alkylene, optionally substituted C 1-6 alkylene, optionally substituted C 2-6 alkylene, optionally substituted C 3-6 alkylene, optionally substituted C 4-6 alkylene, optionally substituted C 4-5 alkylene, or optionally substituted C 3-4 alkylene. In some embodiments, L is optionally substituted C 1 alkylene. In some embodiments, L is optionally substituted C 2 alkylene. In some embodiments, L is optionally substituted C 3 alkylene. In some embodiments, L is optionally substituted C 4 alkylene. In some embodiments, L is optionally substituted C 5 alkylene. In some embodiments, L is optionally substituted C 6 alkylene. In some embodiments, L is optionally substituted C 7 alkylene. In some embodiments, L is optionally substituted C 8 alkylene. In some embodiments, L is optionally substituted C 9 alkylene. In some embodiments, L is optionally substituted C 10 alkylene. In some embodiments, L is —CH 2 —. In some embodiments, L is —(CH 2 ) 2 —. In some embodiments, L is —(CH 2 ) 3 —. In some embodiments, L is —(CH 2 ) 4 —. In some embodiments, L is —(CH 2 ) 5 —. In some embodiments, L is —(CH 2 ) 6 —. In some embodiments, L is —(CH 2 ) 7 —. In some embodiments, L is —(CH 2 ) 8 —. In some embodiments, L is —(CH 2 ) 9 —. In some embodiments, L is —(CH 2 ) 10 —.

In certain embodiments, L is an optionally substituted alkenylene, e.g., optionally substituted C 2-50 alkenylene, optionally substituted C 2-40 alkenylene, optionally substituted C 2-30 alkenylene, optionally substituted C 2-20 alkenylene, optionally substituted C 4-20 alkenylene, optionally substituted C 6-20 alkenylene, optionally substituted C 8-20 alkenylene, optionally substituted C 10-20 alkenylene, optionally substituted C 2-6 alkenylene, optionally substituted C 3-6 alkenylene, optionally substituted C 4-6 alkenylene, optionally substituted C 4-5 alkenylene, or optionally substituted C 3-4 alkenylene.

In certain embodiments, L is an optionally substituted alkynylene, e.g., optionally substituted C 2-50 alkynylene, optionally substituted C 2-40 alkynylene, optionally substituted C 2-30 alkynylene, optionally substituted C 2-20 alkynylene, optionally substituted C 4-20 alkynylene, optionally substituted C 6-20 alkynylene, optionally substituted C 8-20 alkynylene, optionally substituted C 10-20 alkynylene, optionally substituted C 2-6 alkynylene, optionally substituted C 3-6 alkynylene, optionally substituted C 4-6 alkynylene, optionally substituted C 4-5 alkynylene, or optionally substituted C 3-4 alkynylene.

In certain embodiments, L is an optionally substituted heteroalkylene; e.g., optionally substituted heteroC 1-50 alkylene, optionally substituted heteroC 1-40 alkylene, optionally substituted heteroC 1-30 alkylene, optionally substituted heteroC 1-20 alkylene, optionally substituted heteroC 4-20 alkylene, optionally substituted heteroC 6-20 alkylene, optionally substituted heteroC 8-20 alkylene, optionally substituted heteroC 10-20 alkylene, optionally substituted heteroC 1-6 alkylene, optionally substituted heteroC 2-6 alkylene, optionally substituted heteroC 3-6 alkylene, optionally substituted heteroC 4-6 alkylene, optionally substituted heteroC 4-5 alkylene, or optionally substituted heteroC 3-4 alkylene. In some embodiments, L is optionally substituted heteroC 2 alkylene. In some embodiments, L is optionally substituted heteroC 3 alkylene. In some embodiments, L is optionally substituted heteroC 4 alkylene. In some embodiments, L is optionally substituted heteroC 5 alkylene. In some embodiments, L is optionally substituted heteroC 6 alkylene. In some embodiments, L is optionally substituted heteroC 7 alkylene. In some embodiments, L is optionally substituted heteroC 8 alkylene. In some embodiments, L is optionally substituted heteroC 9 alkylene. In some embodiments, L is optionally substituted heteroC 10 alkylene.

In certain embodiments, L is an optionally substituted heteroalkenylene, e.g., optionally substituted heteroC 2-50 alkenylene, optionally substituted heteroC 2-40 alkenylene, optionally substituted heteroC 2-30 alkenylene, optionally substituted heteroC 2-20 alkenylene, optionally substituted heteroC 4-20 alkenylene, optionally substituted heteroC 6-20 alkenylene, optionally substituted heteroC 8-20 alkenylene, optionally substituted heteroC 10-20 alkenylene, optionally substituted heteroC 2-6 alkenylene, optionally substituted heteroC 3-6 alkenylene, optionally substituted heteroC 4-6 alkenylene, optionally substituted heteroC 4-5 alkenylene, or optionally substituted heteroC 3-4 alkenylene.

In certain embodiments, L is an optionally substituted heteroalkynylene, e.g., optionally substituted heteroC 2-50 alkynylene, optionally substituted heteroC 2-40 alkynylene, optionally substituted heteroC 2-30 alkynylene, optionally substituted heteroC 2-20 alkynylene, optionally substituted heteroC 4-20 alkynylene, optionally substituted heteroC 6-20 alkynylene, optionally substituted heteroC 8-20 alkynylene, optionally substituted heteroC 10-20 alkynylene, optionally substituted heteroC 2-6 alkynylene, optionally substituted heteroC 3-6 alkynylene, optionally substituted heteroC 4-6 alkynylene, optionally substituted heteroC 4-5 alkynylene, or optionally substituted heteroC 3-4 alkynylene.

In certain embodiments, L is an optionally substituted carbocyclylene, e.g., optionally substituted C 3-10 carbocyclylene, optionally substituted C 5-8 carbocyclylene, optionally substituted C 5-6 carbocyclylene, optionally substituted C 5 carbocyclylene, or optionally substituted C 6 carbocyclylene.

In certain embodiments, L is an optionally substituted heterocyclylene, e.g., optionally substituted 3-14 membered heterocyclylene, optionally substituted 3-10 membered heterocyclylene, optionally substituted 5-8 membered heterocyclylene, optionally substituted 5-6 membered heterocyclylene, optionally substituted 5-membered heterocyclylene, or optionally substituted 6-membered heterocyclylene.

In certain embodiments, L is an optionally substituted arylene, e.g., optionally substituted phenylene. In some embodiments, L is optionally substituted phenylene. In some embodiments, L is substituted phenylene. In some embodiments, L is unsubstituted phenylene.

In certain embodiments, L is an optionally substituted heteroarylene, e.g., optionally substituted 5-14 membered heteroarylene, optionally substituted 5-10 membered heteroarylene, optionally substituted 5-6 membered heteroarylene, optionally substituted 5-membered heteroarylene, or optionally substituted 6-membered heteroarylene.

In certain embodiments, wherein L is an optionally substituted alkylene group, the group of formula (iv) is a group of the formula

wherein q is an integer between 1 and 50, inclusive, and each of R 6 and R 7 is independently as defined above and described herein.

In certain embodiments, q is an integer between 1 and 40, inclusive. In certain embodiments, q is an integer between 1 and 30, inclusive. In certain embodiments, q is an integer between 1 and 20, inclusive. In certain embodiments, q is an integer between 1 and 10, inclusive. In certain embodiments, q is an integer between 4 and 20, inclusive. In certain embodiments, q is an integer between 6 and 20, inclusive. In certain embodiments, q is an integer between 2 and 10, inclusive. In certain embodiments, q is an integer between 2 and 9, inclusive. In certain embodiments, q is an integer between 2 and 8, inclusive. In certain embodiments, q is an integer between 2 and 7, inclusive. In certain embodiments, q is an integer between 2 and 6, inclusive. In certain embodiments, q is an integer between 2 and 5, inclusive. In certain embodiments, q is an integer between 2 and 4, inclusive. In certain embodiments, q is an integer between 3 and 10, inclusive. In certain embodiments, q is an integer between 3 and 8, inclusive. In certain embodiments, q is an integer between 3 and 7, inclusive. In certain embodiments, q is an integer between 3 and 6, inclusive. In certain embodiments, q is an integer between 3 and 5, inclusive. In certain embodiments, q is 3 or 4. In certain embodiments, q is an integer between 3 and 9, inclusive. In certain embodiments, q is an integer between 8 and 20, inclusive. In certain embodiments, q is 1. In certain embodiments, q is 2. In certain embodiments, q is 3. In certain embodiments, q is 4. In certain embodiments, q is 5. In certain embodiments, q is 6. In certain embodiments, q is 7. In certain embodiments, q is 8. In certain embodiments, q is 9. In certain embodiments, q is 10.

As generally defined above, each R 6 is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, a nitrogen protecting group, or a group of formula (i), (ii) or (iii).

In some embodiments, R 6 is hydrogen.

In some embodiments, R 6 is optionally substituted alkyl. In some embodiments, R 6 is optionally substituted C 2-50 alkyl. In some embodiments, R 6 is optionally substituted C 2-40 alkyl. In some embodiments, R 6 is optionally substituted C 2-30 alkyl. In some embodiments, R 6 is optionally substituted C 2-20 alkyl. In some embodiments, R 6 is optionally substituted C 2-19 alkyl. In some embodiments, R 6 is optionally substituted C 2-18 alkyl. In some embodiments, R 6 is optionally substituted C 2-17 alkyl. In some embodiments, R 6 is optionally substituted C 2-16 alkyl. In some embodiments, R 6 is optionally substituted C 2-15 alkyl. In some embodiments, R 6 is optionally substituted C 2-14 alkyl. In some embodiments, R 6 is optionally substituted C 2-13 alkyl. In some embodiments, R 6 is optionally substituted C 2-12 alkyl. In some embodiments, R 6 is optionally substituted C 2-11 alkyl. In some embodiments, R 6 is optionally substituted C 2-10 alkyl. In some embodiments, R 6 is optionally substituted C 2-9 alkyl. In some embodiments, R 6 is optionally substituted C 2-8 alkyl. In some embodiments, R 6 is optionally substituted C 2-7 alkyl. In some embodiments, R 6 is optionally substituted C 2-6 alkyl.

In some embodiments, R 6 is optionally substituted C 4-50 alkyl. In some embodiments, R 6 is optionally substituted C 4-40 alkyl. In some embodiments, R 6 is optionally substituted C 4-30 alkyl. In some embodiments, R 6 is optionally substituted C 4-20 alkyl. In some embodiments, R 6 is optionally substituted C 4-19 alkyl. In some embodiments, R 6 is optionally substituted C 4-18 alkyl. In some embodiments, R 6 is optionally substituted C 4-17 alkyl. In some embodiments, R 6 is optionally substituted C 4-16 alkyl. In some embodiments, R 6 is optionally substituted C 4-15 alkyl. In some embodiments, R 6 is optionally substituted C 4-14 alkyl. In some embodiments, R 6 is optionally substituted C 4-13 alkyl. In some embodiments, R 6 is optionally substituted C 4-12 alkyl. In some embodiments, R 6 is optionally substituted C 4-11 alkyl. In some embodiments, R 6 is optionally substituted C 4-100 alkyl. In some embodiments, R 6 is optionally substituted C 4-9 alkyl. In some embodiments, R 6 is optionally substituted C 4-8 alkyl. In some embodiments, R 6 is optionally substituted C 4-7 alkyl. In some embodiments, R 6 is optionally substituted C 4-6 alkyl.

In some embodiments, R 6 is optionally substituted C 6-50 alkyl. In some embodiments, R 6 is optionally substituted C 6-40 alkyl. In some embodiments, R 6 is optionally substituted C 6-30 alkyl. In some embodiments, R 6 is optionally substituted C 6-20 alkyl. In some embodiments, R 6 is optionally substituted C 6-19 alkyl. In some embodiments, R 6 is optionally substituted C 6-18 alkyl. In some embodiments, R 6 is optionally substituted C 6-17 alkyl. In some embodiments, R 6 is optionally substituted C 6-16 alkyl. In some embodiments, R 6 is optionally substituted C 6-15 alkyl. In some embodiments, R 6 is optionally substituted C 6-14 alkyl. In some embodiments, R 6 is optionally substituted C 6-13 alkyl. In some embodiments, R 6 is optionally substituted C 6-12 alkyl. In some embodiments, R 6 is optionally substituted C 6-11 alkyl. In some embodiments, R 6 is optionally substituted C 6-10 alkyl. In some embodiments, R 6 is optionally substituted C 6-9 alkyl. In some embodiments, R 6 is optionally substituted C 6-8 alkyl. In some embodiments, R 6 is optionally substituted C 6-7 alkyl.

In some embodiments, R 6 is optionally substituted C 8-50 alkyl. In some embodiments, R 6 is optionally substituted C 8-40 alkyl. In some embodiments, R 6 is optionally substituted C 8-30 alkyl. In some embodiments, R 6 is optionally substituted C 8-20 alkyl. In some embodiments, R 6 is optionally substituted C 8-19 alkyl. In some embodiments, R 6 is optionally substituted C 8-18 alkyl. In some embodiments, R 6 is optionally substituted C 8-17 alkyl. In some embodiments, R 6 is optionally substituted C 8-16 alkyl. In some embodiments, R 6 is optionally substituted C 8-15 alkyl. In some embodiments, R 6 is optionally substituted C 8-14 alkyl. In some embodiments, R 6 is optionally substituted C 8-13 alkyl. In some embodiments, R 6 is optionally substituted C 8-12 alkyl. In some embodiments, R 6 is optionally substituted C 8-11 alkyl. In some embodiments, R 6 is optionally substituted C 8-10 alkyl. In some embodiments, R 6 is optionally substituted C 8-9 alkyl.

In some embodiments, R 6 is optionally substituted C 9-50 alkyl. In some embodiments, R 6 is optionally substituted C 9-40 alkyl. In some embodiments, R 6 is optionally substituted C 9-30 alkyl. In some embodiments, R 6 is optionally substituted C 9-20 alkyl. In some embodiments, R 6 is optionally substituted C 9-19 alkyl. In some embodiments, R 6 is optionally substituted C 9-18 alkyl. In some embodiments, R 6 is optionally substituted C 9-17 alkyl. In some embodiments, R 6 is optionally substituted C 9-16 alkyl. In some embodiments, R 6 is optionally substituted C 9-15 alkyl. In some embodiments, R 6 is optionally substituted C 9-14 alkyl. In some embodiments, R 6 is optionally substituted C 9-13 alkyl. In some embodiments, R 6 is optionally substituted C 9-12 alkyl. In some embodiments, R 6 is optionally substituted C 9-11 alkyl. In some embodiments, R 6 is optionally substituted C 9-10 alkyl.

In some embodiments, R 6 is optionally substituted C 10-50 alkyl. In some embodiments, R 6 is optionally substituted C 10-40 alkyl. In some embodiments, R 6 is optionally substituted C 10-30 alkyl. In some embodiments, R 6 is optionally substituted C 10-20 alkyl. In some embodiments, R 6 is optionally substituted C 10-19 alkyl. In some embodiments, R 6 is optionally substituted C 10-18 alkyl. In some embodiments, R 6 is optionally substituted C 10-17 alkyl. In some embodiments, R 6 is optionally substituted C 10-16 alkyl. In some embodiments, R 6 is optionally substituted C 10-15 alkyl. In some embodiments, R 6 is optionally substituted C 10-14 alkyl. In some embodiments, R 6 is optionally substituted C 10-13 alkyl. In some embodiments, R 6 is optionally substituted C 10-12 alkyl. In some embodiments, R 6 is optionally substituted C 10-11 alkyl.

In some embodiments, R 6 is optionally substituted C 11-50 alkyl. In some embodiments, R 6 is optionally substituted C 11-40 alkyl. In some embodiments, R 6 is optionally substituted C 11-30 alkyl. In some embodiments, R 6 is optionally substituted C 11-20 alkyl. In some embodiments, R 6 is optionally substituted C 11-19 alkyl. In some embodiments, R 6 is optionally substituted C 11-18 alkyl. In some embodiments, R 6 is optionally substituted C 11-17 alkyl. In some embodiments, R 6 is optionally substituted C 11-16 alkyl. In some embodiments, R 6 is optionally substituted C 11-15 alkyl. In some embodiments, R 6 is optionally substituted C 11-14 alkyl. In some embodiments, R 6 is optionally substituted C 11-13 alkyl. In some embodiments, R 6 is optionally substituted C 11-12 alkyl.

In some embodiments, R 6 is optionally substituted C 12-50 alkyl. In some embodiments, R 6 is optionally substituted C 12-40 alkyl. In some embodiments, R 6 is optionally substituted C 12-30 alkyl. In some embodiments, R 6 is optionally substituted C 12-20 alkyl. In some embodiments, R 6 is optionally substituted C 12-19 alkyl. In some embodiments, R 6 is optionally substituted C 12-18 alkyl. In some embodiments, R 6 is optionally substituted C 12-17 alkyl. In some embodiments, R 6 is optionally substituted C 12-16 alkyl. In some embodiments, R 6 is optionally substituted C 12-15 alkyl. In some embodiments, R 6 is optionally substituted C 12-14 alkyl. In some embodiments, R 6 is optionally substituted C 12-13 alkyl.

In some embodiments, R 6 is optionally substituted C 6 alkyl. In some embodiments, R 6 is optionally substituted C 7 alkyl. In some embodiments, R 6 is optionally substituted C 8 alkyl. In some embodiments, R 6 is optionally substituted C 9 alkyl. In some embodiments, R 6 is optionally substituted C 10 alkyl. In some embodiments, R 6 is optionally substituted C 11 alkyl. In some embodiments, R 6 is optionally substituted C 12 alkyl. In some embodiments, R 6 is optionally substituted C 13 alkyl. In some embodiments, R 6 is optionally substituted C 14 alkyl. In some embodiments, R 6 is optionally substituted C 15 alkyl. In some embodiments, R 6 is optionally substituted C 16 alkyl. In some embodiments, R 6 is optionally substituted C 17 alkyl. In some embodiments, R 6 is optionally substituted C 18 alkyl. In some embodiments, R 6 is optionally substituted C 19 alkyl. In some embodiments, R 6 is optionally substituted C 20 alkyl.

In some embodiments, for example, in any of the above embodiments, R 6 is a substituted alkyl group. In some embodiments, R 6 is an unsubstituted alkyl group. In some embodiments, R 6 is an optionally substituted straight-chain alkyl group. In some embodiments, R 6 is a substituted straight-chain alkyl group. In some embodiments, R 6 is an unsubstituted straight-chain alkyl group. In some embodiments, R 6 is an optionally substituted branched alkyl group. In some embodiments, R 6 is a substituted branched alkyl group. In some embodiments, R 6 is an unsubstituted branched alkyl group.

In some embodiments, R 6 is optionally substituted alkenyl. In some embodiments, R 6 is optionally substituted C 2-50 alkenyl. In some embodiments, R 6 is optionally substituted C 2-40 alkenyl. In some embodiments, R 6 is optionally substituted C 2-30 alkenyl. In some embodiments, R 6 is optionally substituted C 2-20 alkenyl. In some embodiments, R 6 is optionally substituted C 2-19 alkenyl. In some embodiments, R 6 is optionally substituted C 2-18 alkenyl. In some embodiments, R 6 is optionally substituted C 2-17 alkenyl. In some embodiments, R 6 is optionally substituted C 2-16 alkenyl. In some embodiments, R 6 is optionally substituted C 2-15 alkenyl. In some embodiments, R 6 is optionally substituted C 2-14 alkenyl. In some embodiments, R 6 is optionally substituted C 2-13 alkenyl. In some embodiments, R 6 is optionally substituted C 2-12 alkenyl. In some embodiments, R 6 is optionally substituted C 2-11 alkenyl. In some embodiments, R 6 is optionally substituted C 2-10 alkenyl. In some embodiments, R 6 is optionally substituted C 2-9 alkenyl. In some embodiments, R 6 is optionally substituted C 2-8 alkenyl. In some embodiments, R 6 is optionally substituted C 2-7 alkenyl. In some embodiments, R 6 is optionally substituted C 2-6 alkenyl.

In some embodiments, R 6 is optionally substituted C 4-50 alkenyl. In some embodiments, R 6 is optionally substituted C 4-40 alkenyl. In some embodiments, R 6 is optionally substituted C 4-30 alkenyl. In some embodiments, R 6 is optionally substituted C 4-20 alkenyl. In some embodiments, R 6 is optionally substituted C 4-19 alkenyl. In some embodiments, R 6 is optionally substituted C 4-18 alkenyl. In some embodiments, R 6 is optionally substituted C 4-17 alkenyl. In some embodiments, R 6 is optionally substituted C 4-16 alkenyl. In some embodiments, R 6 is optionally substituted C 4-15 alkenyl. In some embodiments, R 6 is optionally substituted C 4-14 alkenyl. In some embodiments, R 6 is optionally substituted C 4-13 alkenyl. In some embodiments, R 6 is optionally substituted C 4-12 alkenyl. In some embodiments, R 6 is optionally substituted C 4-11 alkenyl. In some embodiments, R 6 is optionally substituted C 4-10 alkenyl. In some embodiments, R 6 is optionally substituted C 4-9 alkenyl. In some embodiments, R 6 is optionally substituted C 4-8 alkenyl. In some embodiments, R 6 is optionally substituted C 4-7 alkenyl. In some embodiments, R 6 is optionally substituted C 4-6 alkenyl.

In some embodiments, R 6 is optionally substituted C 6-50 alkenyl. In some embodiments, R 6 is optionally substituted C 6-40 alkenyl. In some embodiments, R 6 is optionally substituted C 6-30 alkenyl. In some embodiments, R 6 is optionally substituted C 6-20 alkenyl. In some embodiments, R 6 is optionally substituted C 6-19 alkenyl. In some embodiments, R 6 is optionally substituted C 6-18 alkenyl. In some embodiments, R 6 is optionally substituted C 6-17 alkenyl. In some embodiments, R 6 is optionally substituted C 6-16 alkenyl. In some embodiments, R 6 is optionally substituted C 6-15 alkenyl. In some embodiments, R 6 is optionally substituted C 6-14 alkenyl. In some embodiments, R 6 is optionally substituted C 6-13 alkenyl. In some embodiments, R 6 is optionally substituted C 6-12 alkenyl. In some embodiments, R 6 is optionally substituted C 6-11 alkenyl. In some embodiments, R 6 is optionally substituted C 6-10 alkenyl. In some embodiments, R 6 is optionally substituted C 6-9 alkenyl. In some embodiments, R 6 is optionally substituted C 6-8 alkenyl. In some embodiments, R 6 is optionally substituted C 6-7 alkenyl.

In some embodiments, R 6 is optionally substituted C 8-50 alkenyl. In some embodiments, R 6 is optionally substituted C 8-40 alkenyl. In some embodiments, R 6 is optionally substituted C 8-30 alkenyl. In some embodiments, R 6 is optionally substituted C 8-20 alkenyl. In some embodiments, R 6 is optionally substituted C 8-19 alkenyl. In some embodiments, R 6 is optionally substituted C 8-18 alkenyl. In some embodiments, R 6 is optionally substituted C 8-17 alkenyl. In some embodiments, R 6 is optionally substituted C 8-16 alkenyl. In some embodiments, R 6 is optionally substituted C 8-15 alkenyl. In some embodiments, R 6 is optionally substituted C 8-14 alkenyl. In some embodiments, R 6 is optionally substituted C 8-13 alkenyl. In some embodiments, R 6 is optionally substituted C 8-12 alkenyl. In some embodiments, R 6 is optionally substituted C 8-11 alkenyl. In some embodiments, R 6 is optionally substituted C 8-10 alkenyl. In some embodiments, R 6 is optionally substituted C 8-9 alkenyl.

In some embodiments, R 6 is optionally substituted C 9-50 alkenyl. In some embodiments, R 6 is optionally substituted C 9-40 alkenyl. In some embodiments, R 6 is optionally substituted C 9-30 alkenyl. In some embodiments, R 6 is optionally substituted C 9-20 alkenyl. In some embodiments, R 6 is optionally substituted C 9-19 alkenyl. In some embodiments, R 6 is optionally substituted C 9-18 alkenyl. In some embodiments, R 6 is optionally substituted C 9-17 alkenyl. In some embodiments, R 6 is optionally substituted C 9-16 alkenyl. In some embodiments, R 6 is optionally substituted C 9-15 alkenyl. In some embodiments, R 6 is optionally substituted C 9-14 alkenyl. In some embodiments, R 6 is optionally substituted C 9-13 alkenyl. In some embodiments, R 6 is optionally substituted C 9-12 alkenyl. In some embodiments, R 6 is optionally substituted C 9-11 alkenyl. In some embodiments, R 6 is optionally substituted C 9-10 alkenyl.

In some embodiments, R 6 is optionally substituted C 10-50 alkenyl. In some embodiments, R 6 is optionally substituted C 10-40 alkenyl. In some embodiments, R 6 is optionally substituted C 10-30 alkenyl. In some embodiments, R 6 is optionally substituted C 10-20 alkenyl. In some embodiments, R 6 is optionally substituted C 10-19 alkenyl. In some embodiments, R 6 is optionally substituted C 10-18 alkenyl. In some embodiments, R 6 is optionally substituted C 10-17 alkenyl. In some embodiments, R 6 is optionally substituted C 10-16 alkenyl. In some embodiments, R 6 is optionally substituted C 10-15 alkenyl. In some embodiments, R 6 is optionally substituted C 10-14 alkenyl. In some embodiments, R 6 is optionally substituted C 10-13 alkenyl. In some embodiments, R 6 is optionally substituted C 10-12 alkenyl. In some embodiments, R 6 is optionally substituted C 10-11 alkenyl.

In some embodiments, R 6 is optionally substituted C 11-50 alkenyl. In some embodiments, R 6 is optionally substituted C 11-40 alkenyl. In some embodiments, R 6 is optionally substituted C 11-30 alkenyl. In some embodiments, R 6 is optionally substituted C 11-20 alkenyl. In some embodiments, R 6 is optionally substituted C 11-19 alkenyl. In some embodiments, R 6 is optionally substituted C 1-18 alkenyl. In some embodiments, R 6 is optionally substituted C 1-17 alkenyl. In some embodiments, R 6 is optionally substituted C 11-16 alkenyl. In some embodiments, R 6 is optionally substituted C 11-15 alkenyl. In some embodiments, R 6 is optionally substituted C 11-14 alkenyl. In some embodiments, R 6 is optionally substituted C 11-13 alkenyl. In some embodiments, R 6 is optionally substituted C 11-12 alkenyl.

In some embodiments, R 6 is optionally substituted C 12-50 alkenyl. In some embodiments, R 6 is optionally substituted C 12-40 alkenyl. In some embodiments, R 6 is optionally substituted C 12-30 alkenyl. In some embodiments, R 6 is optionally substituted C 12-20 alkenyl. In some embodiments, R 6 is optionally substituted C 12-19 alkenyl. In some embodiments, R 6 is optionally substituted C 12-18 alkenyl. In some embodiments, R 6 is optionally substituted C 12-17 alkenyl. In some embodiments, R 6 is optionally substituted C 12-16 alkenyl. In some embodiments, R 6 is optionally substituted C 12-15 alkenyl. In some embodiments, R 6 is optionally substituted C 12-14 alkenyl. In some embodiments, R 6 is optionally substituted C 12-13 alkenyl.

In some embodiments, R 6 is optionally substituted C 6 alkenyl. In some embodiments, R 6 is optionally substituted C 7 alkenyl. In some embodiments, R 6 is optionally substituted C 8 alkenyl. In some embodiments, R 6 is optionally substituted C 9 alkenyl. In some embodiments, R 6 is optionally substituted C 10 alkenyl. In some embodiments, R 6 is optionally substituted C 11 alkenyl. In some embodiments, R 6 is optionally substituted C 12 alkenyl. In some embodiments, R 6 is optionally substituted C 13 alkenyl. In some embodiments, R 6 is optionally substituted C 14 alkenyl. In some embodiments, R 6 is optionally substituted C 15 alkenyl. In some embodiments, R 6 is optionally substituted C 16 alkenyl. In some embodiments, R 6 is optionally substituted C 17 alkenyl. In some embodiments, R 6 is optionally substituted C 18 alkenyl. In some embodiments, R 6 is optionally substituted C 19 alkenyl. In some embodiments, R 6 is optionally substituted C 20 alkenyl.

In some embodiments, for example, in any of the above embodiments, R 6 is a substituted alkenyl group. In some embodiments, R 6 is an unsubstituted alkenyl group. In some embodiments, R 6 is an optionally substituted straight-chain alkenyl group. In some embodiments, R 6 is a substituted straight-chain alkenyl group. In some embodiments, R 6 is an unsubstituted straight-chain alkenyl group. In some embodiments, R 6 is an optionally substituted branched alkenyl group. In some embodiments, R 6 is a substituted branched alkenyl group. In some embodiments, R 6 is an unsubstituted branched alkenyl group.

In some embodiments, R 6 is optionally substituted alkynyl. In some embodiments, R 6 is optionally substituted C 2-50 alkynyl. In some embodiments, R 6 is optionally substituted C 2-40 alkynyl. In some embodiments, R 6 is optionally substituted C 2-30 alkynyl. In some embodiments, R 6 is optionally substituted C 2-20 alkynyl. In some embodiments, R 6 is optionally substituted C 2-19 alkynyl. In some embodiments, R 6 is optionally substituted C 2-18 alkynyl. In some embodiments, R 6 is optionally substituted C 2-17 alkynyl. In some embodiments, R 6 is optionally substituted C 2-16 alkynyl. In some embodiments, R 6 is optionally substituted C 2-15 alkynyl. In some embodiments, R 6 is optionally substituted C 2-14 alkynyl. In some embodiments, R 6 is optionally substituted C 2-13 alkynyl. In some embodiments, R 6 is optionally substituted C 2-12 alkynyl. In some embodiments, R 6 is optionally substituted C 2-11 alkynyl. In some embodiments, R 6 is optionally substituted C 2-10 alkynyl. In some embodiments, R 6 is optionally substituted C 2-9 alkynyl. In some embodiments, R 6 is optionally substituted C 2-8 alkynyl. In some embodiments, R 6 is optionally substituted C 2-7 alkynyl. In some embodiments, R 6 is optionally substituted C 2-6 alkynyl.

In some embodiments, R 6 is optionally substituted C 4-50 alkynyl. In some embodiments, R 6 is optionally substituted C 4-40 alkynyl. In some embodiments, R 6 is optionally substituted C 4-30 alkynyl. In some embodiments, R 6 is optionally substituted C 4-20 alkynyl. In some embodiments, R 6 is optionally substituted C 4-19 alkynyl. In some embodiments, R 6 is optionally substituted C 4-18 alkynyl. In some embodiments, R 6 is optionally substituted C 4-17 alkynyl. In some embodiments, R 6 is optionally substituted C 4-16 alkynyl. In some embodiments, R 6 is optionally substituted C 4-15 alkynyl. In some embodiments, R 6 is optionally substituted C 4-14 alkynyl. In some embodiments, R 6 is optionally substituted C 4-13 alkynyl. In some embodiments, R 6 is optionally substituted C 4-12 alkynyl. In some embodiments, R 6 is optionally substituted C 4-11 alkynyl. In some embodiments, R 6 is optionally substituted C 4-10 alkynyl. In some embodiments, R 6 is optionally substituted C 4-9 alkynyl. In some embodiments, R 6 is optionally substituted C 4-8 alkynyl. In some embodiments, R 6 is optionally substituted C 4-7 alkynyl. In some embodiments, R 6 is optionally substituted C 4-6 alkynyl.

In some embodiments, R 6 is optionally substituted C 6-50 alkynyl. In some embodiments, R 6 is optionally substituted C 6-40 alkynyl. In some embodiments, R 6 is optionally substituted C 6-30 alkynyl. In some embodiments, R 6 is optionally substituted C 6-20 alkynyl. In some embodiments, R 6 is optionally substituted C 6-19 alkynyl. In some embodiments, R 6 is optionally substituted C 6-18 alkynyl. In some embodiments, R 6 is optionally substituted C 6-17 alkynyl. In some embodiments, R 6 is optionally substituted C 6-16 alkynyl. In some embodiments, R 6 is optionally substituted C 6-15 alkynyl. In some embodiments, R 6 is optionally substituted C 6-14 alkynyl. In some embodiments, R 6 is optionally substituted C 6-13 alkynyl. In some embodiments, R 6 is optionally substituted C 6-12 alkynyl. In some embodiments, R 6 is optionally substituted C 6-11 alkynyl. In some embodiments, R 6 is optionally substituted C 6-10 alkynyl. In some embodiments, R 6 is optionally substituted C 6-9 alkynyl. In some embodiments, R 6 is optionally substituted C 6-8 alkynyl. In some embodiments, R 6 is optionally substituted C 6-7 alkynyl.

In some embodiments, R 6 is optionally substituted C 8-50 alkynyl. In some embodiments, R 6 is optionally substituted C 8-40 alkynyl. In some embodiments, R 6 is optionally substituted C 8-30 alkynyl. In some embodiments, R 6 is optionally substituted C 8-20 alkynyl. In some embodiments, R 6 is optionally substituted C 8-19 alkynyl. In some embodiments, R 6 is optionally substituted C 8-18 alkynyl. In some embodiments, R 6 is optionally substituted C 8-17 alkynyl. In some embodiments, R 6 is optionally substituted C 8-16 alkynyl. In some embodiments, R 6 is optionally substituted C 8-15 alkynyl. In some embodiments, R 6 is optionally substituted C 8-14 alkynyl. In some embodiments, R 6 is optionally substituted C 8-13 alkynyl. In some embodiments, R 6 is optionally substituted C 8-12 alkynyl. In some embodiments, R 6 is optionally substituted C 8-11 alkynyl. In some embodiments, R 6 is optionally substituted C 8-10 alkynyl. In some embodiments, R 6 is optionally substituted C 8-9 alkynyl.

In some embodiments, R 6 is optionally substituted C 9-50 alkynyl. In some embodiments, R 6 is optionally substituted C 9-40 alkynyl. In some embodiments, R 6 is optionally substituted C 9-30 alkynyl. In some embodiments, R 6 is optionally substituted C 9-20 alkynyl. In some embodiments, R 6 is optionally substituted C 9-19 alkynyl. In some embodiments, R 6 is optionally substituted C 9-18 alkynyl. In some embodiments, R 6 is optionally substituted C 9-17 alkynyl. In some embodiments, R 6 is optionally substituted C 9-16 alkynyl. In some embodiments, R 6 is optionally substituted C 9-15 alkynyl. In some embodiments, R 6 is optionally substituted C 9-14 alkynyl. In some embodiments, R 6 is optionally substituted C 9-13 alkynyl. In some embodiments, R 6 is optionally substituted C 9-12 alkynyl. In some embodiments, R 6 is optionally substituted C 9-11 alkynyl. In some embodiments, R 6 is optionally substituted C 9-10 alkynyl.

In some embodiments, R 6 is optionally substituted C 10-50 alkynyl. In some embodiments, R 6 is optionally substituted C 10-40 alkynyl. In some embodiments, R 6 is optionally substituted C 10-30 alkynyl. In some embodiments, R 6 is optionally substituted C 10-20 alkynyl. In some embodiments, R 6 is optionally substituted C 10-19 alkynyl. In some embodiments, R 6 is optionally substituted C 10-18 alkynyl. In some embodiments, R 6 is optionally substituted C 10-17 alkynyl. In some embodiments, R 6 is optionally substituted C 10-16 alkynyl. In some embodiments, R 6 is optionally substituted C 10-15 alkynyl. In some embodiments, R 6 is optionally substituted C 10-14 alkynyl. In some embodiments, R 6 is optionally substituted C 10-13 alkynyl. In some embodiments, R 6 is optionally substituted C 10-12 alkynyl. In some embodiments, R 6 is optionally substituted C 10-11 alkynyl.

In some embodiments, R 6 is optionally substituted C 11-50 alkynyl. In some embodiments, R 6 is optionally substituted C 11-40 alkynyl. In some embodiments, R 6 is optionally substituted C 1-30 alkynyl. In some embodiments, R 6 is optionally substituted C 11-20 alkynyl. In some embodiments, R 6 is optionally substituted C 11-19 alkynyl. In some embodiments, R 6 is optionally substituted C 11-18 alkynyl. In some embodiments, R 6 is optionally substituted C 11-17 alkynyl. In some embodiments, R 6 is optionally substituted C 11-16 alkynyl. In some embodiments, R 6 is optionally substituted C 11-15 alkynyl. In some embodiments, R 6 is optionally substituted C 11-14 alkynyl. In some embodiments, R 6 is optionally substituted C 11-13 alkynyl. In some embodiments, R 6 is optionally substituted C 11-12 alkynyl.

In some embodiments, R 6 is optionally substituted C 12-50 alkynyl. In some embodiments, R 6 is optionally substituted C 12-40 alkynyl. In some embodiments, R 6 is optionally substituted C 12-30 alkynyl. In some embodiments, R 6 is optionally substituted C 12-20 alkynyl. In some embodiments, R 6 is optionally substituted C 12-19 alkynyl. In some embodiments, R 6 is optionally substituted C 12-18 alkynyl. In some embodiments, R 6 is optionally substituted C 12-17 alkynyl. In some embodiments, R 6 is optionally substituted C 12-16 alkynyl. In some embodiments, R 6 is optionally substituted C 12-15 alkynyl. In some embodiments, R 6 is optionally substituted C 12-14 alkynyl. In some embodiments, R 6 is optionally substituted C 12-13 alkynyl.

In some embodiments, R 6 is optionally substituted C 6 alkynyl. In some embodiments, R 6 is optionally substituted C 7 alkynyl. In some embodiments, R 6 is optionally substituted C 8 alkynyl. In some embodiments, R 6 is optionally substituted C 9 alkynyl. In some embodiments, R 6 is optionally substituted C 10 alkynyl. In some embodiments, R 6 is optionally substituted C 11 alkynyl. In some embodiments, R 6 is optionally substituted C 12 alkynyl. In some embodiments, R 6 is optionally substituted C 13 alkynyl. In some embodiments, R 6 is optionally substituted C 14 alkynyl. In some embodiments, R 6 is optionally substituted C 15 alkynyl. In some embodiments, R 6 is optionally substituted C 16 alkynyl. In some embodiments, R 6 is optionally substituted C 17 alkynyl. In some embodiments, R 6 is optionally substituted C 18 alkynyl. In some embodiments, R 6 is optionally substituted C 19 alkynyl. In some embodiments, R 6 is optionally substituted C 20 alkynyl.

In some embodiments, for example, in any of the above embodiments, R 6 is a substituted alkynyl group. In some embodiments, R 6 is an unsubstituted alknyl group. In some embodiments, R 6 is an optionally substituted straight-chain alkynyl group. In some embodiments, R 6 is a substituted straight-chain alkynyl group. In some embodiments, R 6 is an unsubstituted straight-chain alkynyl group. In some embodiments, R 6 is an optionally substituted branched alkynyl group. In some embodiments, R 6 is a substituted branched alkynyl group. In some embodiments, R 6 is an unsubstituted branched alkynyl group.

In some embodiments, R 6 is optionally substituted carbocyclyl. In some embodiments, R 6 is optionally substituted heterocyclyl. In some embodiments, R 6 is optionally substituted aryl. In some embodiments, R 6 is optionally substituted heteroaryl. In some embodiments, R 6 is a nitrogen protecting group.

In some embodiments, R 6 is a group of formula (i). In some embodiments, R 6 is a group of formula (i-a). In some embodiments, R 6 is a group of formula

In some embodiments, R 6 is a group of formula (i-b). In some embodiments, R 6 is a group of formula (ii). In some embodiments, R 6 is a group of formula (iii).

In some embodiments, R 6 is substituted with one or more hydroxyl groups. In some embodiments, R 6 is substituted with one hydroxyl group. In some embodiments, R 6 is substituted with one 2-hydroxyl group (C1 is the carbon atom directly bonded to the nitrogen atom depicted in formula (iv)).

As generally defined above, each R 7 is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, a nitrogen protecting group, or a group of formula (i), (ii) or (iii).

In some embodiments, R 7 is hydrogen.

In some embodiments, R 7 is optionally substituted alkyl. In some embodiments, R 7 is optionally substituted C 2-50 alkyl. In some embodiments, R 7 is optionally substituted C 2-40 alkyl. In some embodiments, R 7 is optionally substituted C 2-30 alkyl. In some embodiments, R 7 is optionally substituted C 2-20 alkyl. In some embodiments, R 7 is optionally substituted C 2-19 alkyl. In some embodiments, R 7 is optionally substituted C 2-18 alkyl. In some embodiments, R 7 is optionally substituted C 2-17 alkyl. In some embodiments, R 7 is optionally substituted C 2-16 alkyl. In some embodiments, R 7 is optionally substituted C 2-15 alkyl. In some embodiments, R 7 is optionally substituted C 2-14 alkyl. In some embodiments, R 7 is optionally substituted C 2-13 alkyl. In some embodiments, R 7 is optionally substituted C 2-12 alkyl. In some embodiments, R 7 is optionally substituted C 2-11 alkyl. In some embodiments, R 7 is optionally substituted C 2-10 alkyl. In some embodiments, R 7 is optionally substituted C 2-9 alkyl. In some embodiments, R 7 is optionally substituted C 2-8 alkyl. In some embodiments, R 7 is optionally substituted C 2-7 alkyl. In some embodiments, R 7 is optionally substituted C 2-6 alkyl.

In some embodiments, R 7 is optionally substituted C 4-50 alkyl. In some embodiments, R 7 is optionally substituted C 4-40 alkyl. In some embodiments, R 7 is optionally substituted C 4-30 alkyl. In some embodiments, R 7 is optionally substituted C 4-20 alkyl. In some embodiments, R 7 is optionally substituted C 4-19 alkyl. In some embodiments, R 7 is optionally substituted C 4-18 alkyl. In some embodiments, R 7 is optionally substituted C 4-17 alkyl. In some embodiments, R 7 is optionally substituted C 4-16 alkyl. In some embodiments, R 7 is optionally substituted C 4-15 alkyl. In some embodiments, R 7 is optionally substituted C 4-14 alkyl. In some embodiments, R 7 is optionally substituted C 4-13 alkyl. In some embodiments, R 7 is optionally substituted C 4-12 alkyl. In some embodiments, R 7 is optionally substituted C 4-11 alkyl. In some embodiments, R 7 is optionally substituted C 4-10 alkyl. In some embodiments, R 7 is optionally substituted C 4-9 alkyl. In some embodiments, R 7 is optionally substituted C 4-8 alkyl. In some embodiments, R 7 is optionally substituted C 4-7 alkyl. In some embodiments, R 7 is optionally substituted C 4-6 alkyl.

In some embodiments, R 7 is optionally substituted C 6-50 alkyl. In some embodiments, R 7 is optionally substituted C 6-40 alkyl. In some embodiments, R 7 is optionally substituted C 6-30 alkyl. In some embodiments, R 7 is optionally substituted C 6-20 alkyl. In some embodiments, R 7 is optionally substituted C 6-19 alkyl. In some embodiments, R 7 is optionally substituted C 6-18 alkyl. In some embodiments, R 7 is optionally substituted C 6-17 alkyl. In some embodiments, R 7 is optionally substituted C 6-16 alkyl. In some embodiments, R 7 is optionally substituted C 6-15 alkyl. In some embodiments, R 7 is optionally substituted C 6-14 alkyl. In some embodiments, R 7 is optionally substituted C 6-13 alkyl. In some embodiments, R 7 is optionally substituted C 6-12 alkyl. In some embodiments, R 7 is optionally substituted C 6-11 alkyl. In some embodiments, R 7 is optionally substituted C 6-10 alkyl. In some embodiments, R 7 is optionally substituted C 6-9 alkyl. In some embodiments, R 7 is optionally substituted C 6-8 alkyl. In some embodiments, R 7 is optionally substituted C 6-7 alkyl.

In some embodiments, R 7 is optionally substituted C 8-50 alkyl. In some embodiments, R 7 is optionally substituted C 8-40 alkyl. In some embodiments, R 7 is optionally substituted C 8-30 alkyl. In some embodiments, R 7 is optionally substituted C 8-20 alkyl. In some embodiments, R 7 is optionally substituted C 8-19 alkyl. In some embodiments, R 7 is optionally substituted C 8-18 alkyl. In some embodiments, R 7 is optionally substituted C 8-17 alkyl. In some embodiments, R 7 is optionally substituted C 8-16 alkyl. In some embodiments, R 7 is optionally substituted C 8-15 alkyl. In some embodiments, R 7 is optionally substituted C 8-14 alkyl. In some embodiments, R 7 is optionally substituted C 8-13 alkyl. In some embodiments, R 7 is optionally substituted C 8-12 alkyl. In some embodiments, R 7 is optionally substituted C 8-11 alkyl. In some embodiments, R 7 is optionally substituted C 8-10 alkyl. In some embodiments, R 7 is optionally substituted C 8-9 alkyl.

In some embodiments, R 7 is optionally substituted C 9-50 alkyl. In some embodiments, R 7 is optionally substituted C 9-40 alkyl. In some embodiments, R 7 is optionally substituted C 9-30 alkyl. In some embodiments, R 7 is optionally substituted C 9-20 alkyl. In some embodiments, R 7 is optionally substituted C 9-19 alkyl. In some embodiments, R 7 is optionally substituted C 9-18 alkyl. In some embodiments, R 7 is optionally substituted C 9-17 alkyl. In some embodiments, R 7 is optionally substituted C 9-16 alkyl. In some embodiments, R 7 is optionally substituted C 9-15 alkyl. In some embodiments, R 7 is optionally substituted C 9-14 alkyl. In some embodiments, R 7 is optionally substituted C 9-13 alkyl. In some embodiments, R 7 is optionally substituted C 9-12 alkyl. In some embodiments, R 7 is optionally substituted C 9-11 alkyl. In some embodiments, R 7 is optionally substituted C 9-10 alkyl.

In some embodiments, R 7 is optionally substituted C 10-50 alkyl. In some embodiments, R 7 is optionally substituted C 10-40 alkyl. In some embodiments, R 7 is optionally substituted C 10-30 alkyl. In some embodiments, R 7 is optionally substituted C 10-20 alkyl. In some embodiments, R 7 is optionally substituted C 10-19 alkyl. In some embodiments, R 7 is optionally substituted C 10-18 alkyl. In some embodiments, R 7 is optionally substituted C 10-17 alkyl. In some embodiments, R 7 is optionally substituted C 10-16 alkyl. In some embodiments, R 7 is optionally substituted C 10-15 alkyl. In some embodiments, R 7 is optionally substituted C 10-14 alkyl. In some embodiments, R 7 is optionally substituted C 10-13 alkyl. In some embodiments, R 7 is optionally substituted C 10-12 alkyl. In some embodiments, R 7 is optionally substituted C 10-11 alkyl.

In some embodiments, R 7 is optionally substituted C 11-50 alkyl. In some embodiments, R 7 is optionally substituted C 11-40 alkyl. In some embodiments, R 7 is optionally substituted C 11-30 alkyl. In some embodiments, R 7 is optionally substituted C 11-20 alkyl. In some embodiments, R 7 is optionally substituted C 11-19 alkyl. In some embodiments, R 7 is optionally substituted C 11-18 alkyl. In some embodiments, R 7 is optionally substituted C 11-17 alkyl. In some embodiments, R 7 is optionally substituted C 11-16 alkyl. In some embodiments, R 7 is optionally substituted C 11-15 alkyl. In some embodiments, R 7 is optionally substituted C 11-14 alkyl. In some embodiments, R 7 is optionally substituted C 11-13 alkyl. In some embodiments, R 7 is optionally substituted C 11-12 alkyl.

In some embodiments, R 7 is optionally substituted C 12-50 alkyl. In some embodiments, R 7 is optionally substituted C 12-40 alkyl. In some embodiments, R 7 is optionally substituted C 12-30 alkyl. In some embodiments, R 7 is optionally substituted C 12-20 alkyl. In some embodiments, R 7 is optionally substituted C 12-19 alkyl. In some embodiments, R 7 is optionally substituted C 12-18 alkyl. In some embodiments, R 7 is optionally substituted C 12-17 alkyl. In some embodiments, R 7 is optionally substituted C 12-16 alkyl. In some embodiments, R 7 is optionally substituted C 12-15 alkyl. In some embodiments, R 7 is optionally substituted C 12-14 alkyl. In some embodiments, R 7 is optionally substituted C 12-13 alkyl.

In some embodiments, R 7 is optionally substituted C 6 alkyl. In some embodiments, R 7 is optionally substituted C 7 alkyl. In some embodiments, R 7 is optionally substituted C 8 alkyl. In some embodiments, R 7 is optionally substituted C 9 alkyl. In some embodiments, R 7 is optionally substituted C 10 alkyl. In some embodiments, R 7 is optionally substituted C 11 alkyl. In some embodiments, R 7 is optionally substituted C 12 alkyl. In some embodiments, R 7 is optionally substituted C 13 alkyl. In some embodiments, R 7 is optionally substituted C 14 alkyl. In some embodiments, R 7 is optionally substituted C 15 alkyl. In some embodiments, R 7 is optionally substituted C 16 alkyl. In some embodiments, R 7 is optionally substituted C 17 alkyl. In some embodiments, R 7 is optionally substituted C 18 alkyl. In some embodiments, R 7 is optionally substituted C 19 alkyl. In some embodiments, R 7 is optionally substituted C 20 alkyl.

In some embodiments, for example, in any of the above embodiments, R 7 is a substituted alkyl group. In some embodiments, R 7 is an unsubstituted alkyl group. In some embodiments, R 7 is an optionally substituted straight-chain alkyl group. In some embodiments, R 7 is a substituted straight-chain alkyl group. In some embodiments, R 7 is an unsubstituted straight-chain alkyl group. In some embodiments, R 7 is an optionally substituted branched alkyl group. In some embodiments, R 7 is a substituted branched alkyl group. In some embodiments, R 7 is an unsubstituted branched alkyl group.

In some embodiments, R 7 is optionally substituted alkenyl. In some embodiments, R 7 is optionally substituted C 2-50 alkenyl. In some embodiments, R 7 is optionally substituted C 2-40 alkenyl. In some embodiments, R 7 is optionally substituted C 2-30 alkenyl. In some embodiments, R 7 is optionally substituted C 2-20 alkenyl. In some embodiments, R 7 is optionally substituted C 2-19 alkenyl. In some embodiments, R 7 is optionally substituted C 2-18 alkenyl. In some embodiments, R 7 is optionally substituted C 2-17 alkenyl. In some embodiments, R 7 is optionally substituted C 2-16 alkenyl. In some embodiments, R 7 is optionally substituted C 2-15 alkenyl. In some embodiments, R 7 is optionally substituted C 2-14 alkenyl. In some embodiments, R 7 is optionally substituted C 2-13 alkenyl. In some embodiments, R 7 is optionally substituted C 2-12 alkenyl. In some embodiments, R 7 is optionally substituted C 2-11 alkenyl. In some embodiments, R 7 is optionally substituted C 2-10 alkenyl. In some embodiments, R 7 is optionally substituted C 2-9 alkenyl. In some embodiments, R 7 is optionally substituted C 2-8 alkenyl. In some embodiments, R 7 is optionally substituted C 2-7 alkenyl. In some embodiments, R 7 is optionally substituted C 2-6 alkenyl.

In some embodiments, R 7 is optionally substituted C 4-50 alkenyl. In some embodiments, R 7 is optionally substituted C 4-40 alkenyl. In some embodiments, R 7 is optionally substituted C 4-30 alkenyl. In some embodiments, R 7 is optionally substituted C 4-20 alkenyl. In some embodiments, R 7 is optionally substituted C 4-19 alkenyl. In some embodiments, R 7 is optionally substituted C 4-18 alkenyl. In some embodiments, R 7 is optionally substituted C 4-17 alkenyl. In some embodiments, R 7 is optionally substituted C 4-16 alkenyl. In some embodiments, R 7 is optionally substituted C 4-15 alkenyl. In some embodiments, R 7 is optionally substituted C 4-14 alkenyl. In some embodiments, R 7 is optionally substituted C 4-13 alkenyl. In some embodiments, R 7 is optionally substituted C 4-12 alkenyl. In some embodiments, R 7 is optionally substituted C 4-11 alkenyl. In some embodiments, R 7 is optionally substituted C 4-10 alkenyl. In some embodiments, R 7 is optionally substituted C 4-9 alkenyl. In some embodiments, R 7 is optionally substituted C 4-8 alkenyl. In some embodiments, R 7 is optionally substituted C 4-7 alkenyl. In some embodiments, R 7 is optionally substituted C 4-6 alkenyl.

In some embodiments, R 7 is optionally substituted C 6-50 alkenyl. In some embodiments, R 7 is optionally substituted C 6-40 alkenyl. In some embodiments, R 7 is optionally substituted C 6-30 alkenyl. In some embodiments, R 7 is optionally substituted C 6-20 alkenyl. In some embodiments, R 7 is optionally substituted C 6-19 alkenyl. In some embodiments, R 7 is optionally substituted C 6-18 alkenyl. In some embodiments, R 7 is optionally substituted C 6-17 alkenyl. In some embodiments, R 7 is optionally substituted C 6-16 alkenyl. In some embodiments, R 7 is optionally substituted C 6-15 alkenyl. In some embodiments, R 7 is optionally substituted C 6-14 alkenyl. In some embodiments, R 7 is optionally substituted C 6-13 alkenyl. In some embodiments, R 7 is optionally substituted C 6-12 alkenyl. In some embodiments, R 7 is optionally substituted C 6-11 alkenyl. In some embodiments, R 7 is optionally substituted C 6-10 alkenyl. In some embodiments, R 7 is optionally substituted C 6-9 alkenyl. In some embodiments, R 7 is optionally substituted C 6-8 alkenyl. In some embodiments, R 7 is optionally substituted C 6-7 alkenyl.

In some embodiments, R 7 is optionally substituted C 8-50 alkenyl. In some embodiments, R 7 is optionally substituted C 8-40 alkenyl. In some embodiments, R 7 is optionally substituted C 8-30 alkenyl. In some embodiments, R 7 is optionally substituted C 8-20 alkenyl. In some embodiments, R 7 is optionally substituted C 8-19 alkenyl. In some embodiments, R 7 is optionally substituted C 8-18 alkenyl. In some embodiments, R 7 is optionally substituted C 8-17 alkenyl. In some embodiments, R 7 is optionally substituted C 8-16 alkenyl. In some embodiments, R 7 is optionally substituted C 8-15 alkenyl. In some embodiments, R 7 is optionally substituted C 8-14 alkenyl. In some embodiments, R 7 is optionally substituted C 8-13 alkenyl. In some embodiments, R 7 is optionally substituted C 8-12 alkenyl. In some embodiments, R 7 is optionally substituted C 8-11 alkenyl. In some embodiments, R 7 is optionally substituted C 8-10 alkenyl. In some embodiments, R 7 is optionally substituted C 8-9 alkenyl.

In some embodiments, R 7 is optionally substituted C 9-50 alkenyl. In some embodiments, R 7 is optionally substituted C 9-40 alkenyl. In some embodiments, R 7 is optionally substituted C 9-30 alkenyl. In some embodiments, R 7 is optionally substituted C 9-20 alkenyl. In some embodiments, R 7 is optionally substituted C 9-19 alkenyl. In some embodiments, R 7 is optionally substituted C 9-18 alkenyl. In some embodiments, R 7 is optionally substituted C 9-17 alkenyl. In some embodiments, R 7 is optionally substituted C 9-16 alkenyl. In some embodiments, R 7 is optionally substituted C 9-15 alkenyl. In some embodiments, R 7 is optionally substituted C 9-14 alkenyl. In some embodiments, R 7 is optionally substituted C 9-13 alkenyl. In some embodiments, R 7 is optionally substituted C 9-12 alkenyl. In some embodiments, R 7 is optionally substituted C 9-11 alkenyl. In some embodiments, R 7 is optionally substituted C 9-10 alkenyl.

In some embodiments, R 7 is optionally substituted C 10-50 alkenyl. In some embodiments, R 7 is optionally substituted C 10-40 alkenyl. In some embodiments, R 7 is optionally substituted C 10-30 alkenyl. In some embodiments, R 7 is optionally substituted C 10-20 alkenyl. In some embodiments, R 7 is optionally substituted C 10-19 alkenyl. In some embodiments, R 7 is optionally substituted C 10-18 alkenyl. In some embodiments, R 7 is optionally substituted C 10-17 alkenyl. In some embodiments, R 7 is optionally substituted C 10-16 alkenyl. In some embodiments, R 7 is optionally substituted C 10-15 alkenyl. In some embodiments, R 7 is optionally substituted C 10-14 alkenyl. In some embodiments, R 7 is optionally substituted C 10-13 alkenyl. In some embodiments, R 7 is optionally substituted C 10-12 alkenyl. In some embodiments, R 7 is optionally substituted C 10-11 alkenyl.

In some embodiments, R 7 is optionally substituted C 11-50 alkenyl. In some embodiments, R 7 is optionally substituted C 11-40 alkenyl. In some embodiments, R 7 is optionally substituted C 11-30 alkenyl. In some embodiments, R 7 is optionally substituted C 11-20 alkenyl. In some embodiments, R 7 is optionally substituted C 11-19 alkenyl. In some embodiments, R 7 is optionally substituted C 11-18 alkenyl. In some embodiments, R 7 is optionally substituted C 11-17 alkenyl. In some embodiments, R 7 is optionally substituted C 11-16 alkenyl. In some embodiments, R 7 is optionally substituted C 11-15 alkenyl. In some embodiments, R 7 is optionally substituted C 11-14 alkenyl. In some embodiments, R 7 is optionally substituted C 11-13 alkenyl. In some embodiments, R 7 is optionally substituted C 11-12 alkenyl.

In some embodiments, R 7 is optionally substituted C 12-50 alkenyl. In some embodiments, R 7 is optionally substituted C 12-40 alkenyl. In some embodiments, R 7 is optionally substituted C 12-30 alkenyl. In some embodiments, R 7 is optionally substituted C 12-20 alkenyl. In some embodiments, R 7 is optionally substituted C 12-19 alkenyl. In some embodiments, R 7 is optionally substituted C 12-18 alkenyl. In some embodiments, R 7 is optionally substituted C 12-17 alkenyl. In some embodiments, R 7 is optionally substituted C 12-16 alkenyl. In some embodiments, R 7 is optionally substituted C 12-15 alkenyl. In some embodiments, R 7 is optionally substituted C 12-14 alkenyl. In some embodiments, R 7 is optionally substituted C 12-13 alkenyl.

In some embodiments, R 7 is optionally substituted C 6 alkenyl. In some embodiments, R 7 is optionally substituted C 7 alkenyl. In some embodiments, R 7 is optionally substituted C 8 alkenyl. In some embodiments, R 7 is optionally substituted C 9 alkenyl. In some embodiments, R 7 is optionally substituted C 10 alkenyl. In some embodiments, R 7 is optionally substituted C 11 alkenyl. In some embodiments, R 7 is optionally substituted C 12 alkenyl. In some embodiments, R 7 is optionally substituted C 13 alkenyl. In some embodiments, R 7 is optionally substituted C 14 alkenyl. In some embodiments, R 7 is optionally substituted C 15 alkenyl. In some embodiments, R 7 is optionally substituted C 16 alkenyl. In some embodiments, R 7 is optionally substituted C 17 alkenyl. In some embodiments, R 7 is optionally substituted C 18 alkenyl. In some embodiments, R 7 is optionally substituted C 19 alkenyl. In some embodiments, R 7 is optionally substituted C 20 alkenyl.

In some embodiments, for example, in any of the above embodiments, R 7 is a substituted alkenyl group. In some embodiments, R 7 is an unsubstituted alkenyl group. In some embodiments, R 7 is an optionally substituted straight-chain alkenyl group. In some embodiments, R 7 is a substituted straight-chain alkenyl group. In some embodiments, R 7 is an unsubstituted straight-chain alkenyl group. In some embodiments, R 7 is an optionally substituted branched alkenyl group. In some embodiments, R 7 is a substituted branched alkenyl group. In some embodiments, R 7 is an unsubstituted branched alkenyl group.

In some embodiments, R 7 is optionally substituted alkynyl. In some embodiments, R 7 is optionally substituted C 2-50 alkynyl. In some embodiments, R 7 is optionally substituted C 2-40 alkynyl. In some embodiments, R 7 is optionally substituted C 2-30 alkynyl. In some embodiments, R 7 is optionally substituted C 2-20 alkynyl. In some embodiments, R 7 is optionally substituted C 2-19 alkynyl. In some embodiments, R 7 is optionally substituted C 2-18 alkynyl. In some embodiments, R 7 is optionally substituted C 2-17 alkynyl. In some embodiments, R 7 is optionally substituted C 2-16 alkynyl. In some embodiments, R 7 is optionally substituted C 2-15 alkynyl. In some embodiments, R 7 is optionally substituted C 2-14 alkynyl. In some embodiments, R 7 is optionally substituted C 2-13 alkynyl. In some embodiments, R 7 is optionally substituted C 2-12 alkynyl. In some embodiments, R 7 is optionally substituted C 2-11 alkynyl. In some embodiments, R 7 is optionally substituted C 2-10 alkynyl. In some embodiments, R 7 is optionally substituted C 2-9 alkynyl. In some embodiments, R 7 is optionally substituted C 2-8 alkynyl. In some embodiments, R 7 is optionally substituted C 2-7 alkynyl. In some embodiments, R 7 is optionally substituted C 2-6 alkynyl.

In some embodiments, R 7 is optionally substituted C 4-50 alkynyl. In some embodiments, R 7 is optionally substituted C 4-40 alkynyl. In some embodiments, R 7 is optionally substituted C 4-30 alkynyl. In some embodiments, R 7 is optionally substituted C 4-20 alkynyl. In some embodiments, R 7 is optionally substituted C 4-19 alkynyl. In some embodiments, R 7 is optionally substituted C 4-18 alkynyl. In some embodiments, R 7 is optionally substituted C 4-17 alkynyl. In some embodiments, R 7 is optionally substituted C 4-16 alkynyl. In some embodiments, R 7 is optionally substituted C 4-15 alkynyl. In some embodiments, R 7 is optionally substituted C 4-14 alkynyl. In some embodiments, R 7 is optionally substituted C 4-13 alkynyl. In some embodiments, R 7 is optionally substituted C 4-12 alkynyl. In some embodiments, R 7 is optionally substituted C 4-11 alkynyl. In some embodiments, R 7 is optionally substituted C 4-10 alkynyl. In some embodiments, R 7 is optionally substituted C 4-9 alkynyl. In some embodiments, R 7 is optionally substituted C 4-8 alkynyl. In some embodiments, R 7 is optionally substituted C 4-7 alkynyl. In some embodiments, R 7 is optionally substituted C 4-6 alkynyl.

In some embodiments, R 7 is optionally substituted C 6-50 alkynyl. In some embodiments, R 7 is optionally substituted C 6-40 alkynyl. In some embodiments, R 7 is optionally substituted C 6-30 alkynyl. In some embodiments, R 7 is optionally substituted C 6-20 alkynyl. In some embodiments, R 7 is optionally substituted C 6-19 alkynyl. In some embodiments, R 7 is optionally substituted C 6-18 alkynyl. In some embodiments, R 7 is optionally substituted C 6-17 alkynyl. In some embodiments, R 7 is optionally substituted C 6-16 alkynyl. In some embodiments, R 7 is optionally substituted C 6-15 alkynyl. In some embodiments, R 7 is optionally substituted C 6-14 alkynyl. In some embodiments, R 7 is optionally substituted C 6-13 alkynyl. In some embodiments, R 7 is optionally substituted C 6-12 alkynyl. In some embodiments, R 7 is optionally substituted C 6-11 alkynyl. In some embodiments, R 7 is optionally substituted C 6-10 alkynyl. In some embodiments, R 7 is optionally substituted C 6-9 alkynyl. In some embodiments, R 7 is optionally substituted C 6-8 alkynyl. In some embodiments, R 7 is optionally substituted C 6-7 alkynyl.

In some embodiments, R 7 is optionally substituted C 8-50 alkynyl. In some embodiments, R 7 is optionally substituted C 8-40 alkynyl. In some embodiments, R 7 is optionally substituted C 8-30 alkynyl. In some embodiments, R 7 is optionally substituted C 8-20 alkynyl. In some embodiments, R 7 is optionally substituted C 8-19 alkynyl. In some embodiments, R 7 is optionally substituted C 8-18 alkynyl. In some embodiments, R 7 is optionally substituted C 8-17 alkynyl. In some embodiments, R 7 is optionally substituted C 8-16 alkynyl. In some embodiments, R 7 is optionally substituted C 8-15 alkynyl. In some embodiments, R 7 is optionally substituted C 8-14 alkynyl. In some embodiments, R 7 is optionally substituted C 8-13 alkynyl. In some embodiments, R 7 is optionally substituted C 8-12 alkynyl. In some embodiments, R 7 is optionally substituted C 8-11 alkynyl. In some embodiments, R 7 is optionally substituted C 8-10 alkynyl. In some embodiments, R 7 is optionally substituted C 8-9 alkynyl.

In some embodiments, R 7 is optionally substituted C 9-50 alkynyl. In some embodiments, R 7 is optionally substituted C 9-40 alkynyl. In some embodiments, R 7 is optionally substituted C 9-30 alkynyl. In some embodiments, R 7 is optionally substituted C 9-20 alkynyl. In some embodiments, R 7 is optionally substituted C 9-19 alkynyl. In some embodiments, R 7 is optionally substituted C 9-18 alkynyl. In some embodiments, R 7 is optionally substituted C 9-17 alkynyl. In some embodiments, R 7 is optionally substituted C 9-16 alkynyl. In some embodiments, R 7 is optionally substituted C 9-15 alkynyl. In some embodiments, R 7 is optionally substituted C 9-14 alkynyl. In some embodiments, R 7 is optionally substituted C 9-13 alkynyl. In some embodiments, R 7 is optionally substituted C 9-12 alkynyl. In some embodiments, R 7 is optionally substituted C 9-11 alkynyl. In some embodiments, R 7 is optionally substituted C 9-10 alkynyl.

In some embodiments, R 7 is optionally substituted C 10-50 alkynyl. In some embodiments, R 7 is optionally substituted C 10-40 alkynyl. In some embodiments, R 7 is optionally substituted C 10-30 alkynyl. In some embodiments, R 7 is optionally substituted C 10-20 alkynyl. In some embodiments, R 7 is optionally substituted C 10-19 alkynyl. In some embodiments, R 7 is optionally substituted C 10-18 alkynyl. In some embodiments, R 7 is optionally substituted C 10-17 alkynyl. In some embodiments, R 7 is optionally substituted C 10-16 alkynyl. In some embodiments, R 7 is optionally substituted C 10-15 alkynyl. In some embodiments, R 7 is optionally substituted C 10-14 alkynyl. In some embodiments, R 7 is optionally substituted C 10-13 alkynyl. In some embodiments, R 7 is optionally substituted C 10-12 alkynyl. In some embodiments, R 7 is optionally substituted C 10-11 alkynyl.

In some embodiments, R 7 is optionally substituted C 11-50 alkynyl. In some embodiments, R 7 is optionally substituted C 11-40 alkynyl. In some embodiments, R 7 is optionally substituted C 11-30 alkynyl. In some embodiments, R 7 is optionally substituted C 11-20 alkynyl. In some embodiments, R 7 is optionally substituted C 11-19 alkynyl. In some embodiments, R 7 is optionally substituted C 11-18 alkynyl. In some embodiments, R 7 is optionally substituted C 11-17 alkynyl. In some embodiments, R 7 is optionally substituted C 11-16 alkynyl. In some embodiments, R 7 is optionally substituted C 11-15 alkynyl. In some embodiments, R 7 is optionally substituted C 11-14 alkynyl. In some embodiments, R 7 is optionally substituted C 11-13 alkynyl. In some embodiments, R 7 is optionally substituted C 11-12 alkynyl.

In some embodiments, R 7 is optionally substituted C 12-50 alkynyl. In some embodiments, R 7 is optionally substituted C 12-40 alkynyl. In some embodiments, R 7 is optionally substituted C 12-30 alkynyl. In some embodiments, R 7 is optionally substituted C 12-20 alkynyl. In some embodiments, R 7 is optionally substituted C 12-19 alkynyl. In some embodiments, R 7 is optionally substituted C 12-18 alkynyl. In some embodiments, R 7 is optionally substituted C 12-17 alkynyl. In some embodiments, R 7 is optionally substituted C 12-16 alkynyl. In some embodiments, R 7 is optionally substituted C 12-15 alkynyl. In some embodiments, R 7 is optionally substituted C 12-14 alkynyl. In some embodiments, R 7 is optionally substituted C 12-13 alkynyl.

In some embodiments, R 7 is optionally substituted C 6 alkynyl. In some embodiments, R 7 is optionally substituted C 7 alkynyl. In some embodiments, R 7 is optionally substituted C 8 alkynyl. In some embodiments, R 7 is optionally substituted C 9 alkynyl. In some embodiments, R 7 is optionally substituted C 10 alkynyl. In some embodiments, R 7 is optionally substituted C 11 alkynyl. In some embodiments, R 7 is optionally substituted C 12 alkynyl. In some embodiments, R 7 is optionally substituted C 13 alkynyl. In some embodiments, R 7 is optionally substituted C 14 alkynyl. In some embodiments, R 7 is optionally substituted C 15 alkynyl. In some embodiments, R 7 is optionally substituted C 16 alkynyl. In some embodiments, R 7 is optionally substituted C 17 alkynyl. In some embodiments, R 7 is optionally substituted C 18 alkynyl. In some embodiments, R 7 is optionally substituted C 19 alkynyl. In some embodiments, R 7 is optionally substituted C 20 alkynyl.

In some embodiments, for example, in any of the above embodiments, R 7 is a substituted alkynyl group. In some embodiments, R 7 is an unsubstituted alkynyl group. In some embodiments, R 7 is an optionally substituted straight-chain alkynyl group. In some embodiments, R 7 is a substituted straight-chain alkynyl group. In some embodiments, R 7 is an unsubstituted straight-chain alkynyl group. In some embodiments, R 7 is an optionally substituted branched alkynyl group. In some embodiments, R 7 is a substituted branched alkynyl group. In some embodiments, R 7 is an unsubstituted branched alkynyl group.

In some embodiments, R 7 is optionally substituted carbocyclyl. In some embodiments, R 7 is optionally substituted heterocyclyl. In some embodiments, R 7 is optionally substituted aryl. In some embodiments, R 7 is optionally substituted heteroaryl. In some embodiments, R 7 is a nitrogen protecting group.

In some embodiments, R 7 is a group of formula (i). In some embodiments, R 7 is a group of formula (i-a). In some embodiments, R 7 is a group of formula

In some embodiments, R 7 is a group of formula (i-b). In some embodiments, R 7 is a group of formula (ii). In some embodiments, R 7 is a group of formula (iii).

In some embodiments, at least one instance of R 6 and R 7 is a group of the formula (i), (ii) or (iii). In some embodiments, each instance of R 6 and R 7 is independently a group of the formula (i), (ii) or (iii). In some embodiments, each instance of R 6 and R 7 is independently a group of the formula (i). In some embodiments, each instance of R 6 and R 7 is independently a group of the formula (i-a). In some embodiments, each instance of R 6 and R 7 is independently a group of the formula (i-b). In some embodiments, each instance of R 6 and R 7 is independently a group of the formula (ii). In some embodiments, each instance of R 6 and R 7 is independently a group of the formula (iii).

In some embodiments, R 6 and R 7 are the same. In some embodiments, R 6 and R 7 are different.

In certain embodiments, both R 6 and R 7 are hydrogen. In certain embodiments, R 6 is hydrogen and R 7 is a group of the formula (i), (ii), or (iii). In certain embodiments, R 6 is hydrogen and R 7 is a group of the formula (i). In certain embodiments, R 6 is hydrogen and R 7 is a group of the formula (ii). In certain embodiments, R 6 is hydrogen and R 7 is a group of the formula (iii). In certain embodiments, each of R 6 and R 7 is independently a group of the formula (i), (ii), or (iii). In certain embodiments, each of R 6 and R 7 is independently a group of the formula (i). In certain embodiments, each of R 6 and R 7 is independently a group of the formula (ii). In certain embodiments, each of R 6 and R 7 is independently a group of the formula (iii). In certain embodiments, R 6 and R 7 are the same group, which is selected from formulas (i), (ii), and (iii). In some embodiments, R 6 and R 7 are the same group of formula (i). In some embodiments, R 6 and R 7 are the same group of formula (i-a). In some embodiments, R 6 and R 7 are the same group of formula (i-al). In some embodiments, R 6 and R 7 are the same group of formula (i-b).

In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is as defined above and described herein. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 1-50 alkyl, optionally substituted C 2-50 alkenyl, optionally substituted C 2-50 alkynyl, optionally substituted heteroC 1-50 alkyl, optionally substituted heteroC 2-50 alkenyl, or optionally substituted heteroC 2-50 alkynyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 5-50 alkyl, optionally substituted C 5-50 alkenyl, optionally substituted C 5-50 alkynyl, optionally substituted heteroC 5-50 alkyl, optionally substituted heteroC 5-50 alkenyl, or optionally substituted heteroC 5-50 alkynyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 5-40 alkyl, optionally substituted C 5-40 alkenyl, optionally substituted C 5-40 alkynyl, optionally substituted heteroC 5-40 alkyl, optionally substituted heteroC 5-40 alkenyl, or optionally substituted heteroC 5-40 alkynyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 5-30 alkyl, optionally substituted C 5-30 alkenyl, optionally substituted C 5-30 alkynyl, optionally substituted heteroC 5-30 alkyl, optionally substituted heteroC 5-30 alkenyl, or optionally substituted heteroC 5-30 alkynyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 5-25 alkyl, optionally substituted C 5-25 alkenyl, optionally substituted C 5-25 alkynyl, optionally substituted heteroC 5-25 alkyl, optionally substituted heteroC 5-25 alkenyl, or optionally substituted heteroC 5-25 alkynyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 5-20 alkyl, optionally substituted C 5-20 alkenyl, optionally substituted C 5-20 alkynyl, optionally substituted heteroC 5-20 alkyl, optionally substituted heteroC 5-20 alkenyl, or optionally substituted heteroC 5-20 alkynyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 5-15 alkyl, optionally substituted C 5-15 alkenyl, optionally substituted C 5-15 alkynyl, optionally substituted heteroC 5-15 alkyl, optionally substituted heteroC 5-15 alkenyl, or optionally substituted heteroC 5-15 alkynyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 5 alkyl, optionally substituted C 5 alkenyl, optionally substituted C 5 alkynyl, optionally substituted heteroC 5 alkyl, optionally substituted heteroC 5 alkenyl, or optionally substituted heteroC 5 alkynyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 6 alkyl, optionally substituted C 6 alkenyl, optionally substituted C 6 alkynyl, optionally substituted heteroC 6 alkyl, optionally substituted heteroC 6 alkenyl, or optionally substituted heteroC 6 alkynyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 7 alkyl, optionally substituted C 7 alkenyl, optionally substituted C 7 alkynyl, optionally substituted heteroC 7 alkyl, optionally substituted heteroC 7 alkenyl, or optionally substituted heteroC 7 alkynyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 8 alkyl, optionally substituted C 8 alkenyl, optionally substituted C 8 alkynyl, optionally substituted heteroC 5 alkyl, optionally substituted heteroC 5 alkenyl, or optionally substituted heteroC 5 alkynyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein K is optionally substituted C 9 alkyl, optionally substituted C 9 alkenyl, optionally substituted C 9 alkynyl, optionally substituted heteroC 9 alkyl, optionally substituted heteroC 9 alkenyl, or optionally substituted heteroC 9 alkynyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 10 alkyl, optionally substituted C 10 alkenyl, optionally substituted C 10 alkynyl, optionally substituted heteroC 10 alkyl, optionally substituted heteroC 10 alkenyl, or optionally substituted heteroC 10 alkynyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C11 alkyl, optionally substituted C 11 alkenyl, optionally substituted C 11 alkynyl, optionally substituted heteroC 11 alkyl, optionally substituted heteroC 11 alkenyl, or optionally substituted heteroC 11 alkynyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 12 alkyl, optionally substituted C 12 alkenyl, optionally substituted C 12 alkynyl, optionally substituted heteroC 12 alkyl, optionally substituted heteroC 12 alkenyl, or optionally substituted heteroC 12 alkynyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 13 alkyl, optionally substituted C 13 alkenyl, optionally substituted C 13 alkynyl, optionally substituted heteroC 13 alkyl, optionally substituted heteroC 13 alkenyl, or optionally substituted heteroC 1-3 alkynyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 14 alkyl, optionally substituted C 14 alkenyl, optionally substituted C 14 alkynyl, optionally substituted heteroC 14 alkyl, optionally substituted heteroC 14 alkenyl, or optionally substituted heteroC 14 alkynyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 15 alkyl, optionally substituted C 15 alkenyl, optionally substituted C 15 alkynyl, optionally substituted heteroC 15 alkyl, optionally substituted heteroC 15 alkenyl, or optionally substituted heteroC 15 alkynyl.

In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is as defined above and described herein. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 1-50 alkyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 5-50 alkyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 5-40 alkyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 5-30 alkyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 5-25 alkyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 5-20 alkyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R is optionally substituted C 5-15 alkyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 5 alkyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 6 alkyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 7 alkyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 8 alkyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 9 alkyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 10 alkyl. In some embodiments, R 6 and R are the same group of formula

wherein R L is optionally substituted C 11 alkyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 12 alkyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 13 alkyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 14 alkyl. In some embodiments, R 6 and R 7 are the same group of formula

wherein R L is optionally substituted C 15 alkyl.

As generally defined above, each occurrence of R A1 is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, an oxygen protecting group when attached to an oxygen atom, a sulfur protecting group when attached to an sulfur atom, a nitrogen protecting group when attached to a nitrogen atom, or two R A1 groups, together with the nitrogen atom to which they are attached, are joined to form an optionally substituted heterocyclic or optionally substituted heteroaryl ring.

In some embodiments, R A1 is hydrogen. In some embodiments, R A1 is optionally substituted alkyl. In some embodiments, R A1 is optionally substituted alkenyl. In some embodiments, R A1 is optionally substituted alkynyl. In some embodiments, R A1 is optionally substituted carbocyclyl. In some embodiments, R A1 is optionally substituted heterocyclyl. In some embodiments, R A1 is optionally substituted aryl. In some embodiments, R A1 is optionally substituted heteroaryl. In some embodiments, R A1 is an oxygen protecting group when attached to an oxygen atom. In some embodiments, R A1 is a sulfur protecting group when attached to a sulfur atom. In some embodiments, R A1 is a nitrogen protecting group when attached to a nitrogen atom. In some embodiments, two R A1 groups, together with the nitrogen atom to which they are attached, are joined to form an optionally substituted heterocyclic or optionally substituted heteroaryl ring.

As generally defined above, each instance of R 2 is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, a nitrogen protecting group, or a group of the formula (i), (ii), or (iii):

wherein each of R′, Y, R P , R L and X is independently as defined above and described herein.

In some embodiments, R 2 is hydrogen. In some embodiments, at least one instance of R 2 is hydrogen. In some embodiments, each instance of R 2 is hydrogen.

In certain embodiments, R 2 is optionally substituted alkyl; e.g., optionally substituted C 1-6 alkyl, optionally substituted C 2-6 alkyl, optionally substituted C 3-6 alkyl, optionally substituted C 4-6 alkyl, optionally substituted C 4-5 alkyl, or optionally substituted C 3-4 alkyl. In certain embodiments, at least one instance of R 2 is optionally substituted alkyl; e.g., optionally substituted C 1-6 alkyl, optionally substituted C 2-6 alkyl, optionally substituted C 3-6 alkyl, optionally substituted C 4-6 alkyl, optionally substituted C 4-5 alkyl, or optionally substituted C 3-4 alkyl.

In certain embodiments, R 2 is optionally substituted alkenyl, e.g., optionally substituted C 2-6 alkenyl, optionally substituted C 3-6 alkenyl, optionally substituted C 4-6 alkenyl, optionally substituted C 4-5 alkenyl, or optionally substituted C 3-4 alkenyl. In certain embodiments, at least one instance of R 2 is optionally substituted alkenyl, e.g., optionally substituted C 2-6 alkenyl, optionally substituted C 3-6 alkenyl, optionally substituted C 4-6 alkenyl, optionally substituted C 4-5 alkenyl, or optionally substituted C 3-4 alkenyl.

In certain embodiments, R 2 is optionally substituted alkynyl, e.g., optionally substituted C 2-6 alkynyl, optionally substituted C 3-6 alkynyl, optionally substituted C 4-6 alkynyl, optionally substituted C 4-5 alkynyl, or optionally substituted C 3-4 alkynyl. In certain embodiments, at least one instance of R 2 is optionally substituted alkynyl, e.g., optionally substituted C 2-6 alkynyl, optionally substituted C 3-6 alkynyl, optionally substituted C 4-6 alkynyl, optionally substituted C 4-5 alkynyl, or optionally substituted C 3-4 alkynyl.

In certain embodiments, R 2 is optionally substituted carbocyclyl, e.g., optionally substituted C 3-10 carbocyclyl, optionally substituted C 5-8 carbocyclyl, optionally substituted C 5-6 carbocyclyl, optionally substituted C 5 carbocyclyl, or optionally substituted C 6 carbocyclyl. In certain embodiments, at least one instance of R 2 is optionally substituted carbocyclyl, e.g., optionally substituted C 3-10 carbocyclyl, optionally substituted C 5-8 carbocyclyl, optionally substituted C 5-6 carbocyclyl, optionally substituted C 5 carbocyclyl, or optionally substituted C 6 carbocyclyl.

In certain embodiments, R 2 is optionally substituted heterocyclyl, e.g., optionally substituted 3-14 membered heterocyclyl, optionally substituted 3-10 membered heterocyclyl, optionally substituted 5-8 membered heterocyclyl, optionally substituted 5-6 membered heterocyclyl, optionally substituted 5-membered heterocyclyl, or optionally substituted 6-membered heterocyclyl. In certain embodiments, at least one instance of R 2 is optionally substituted heterocyclyl, e.g., optionally substituted 3-14 membered heterocyclyl, optionally substituted 3-10 membered heterocyclyl, optionally substituted 5-8 membered heterocyclyl, optionally substituted 5-6 membered heterocyclyl, optionally substituted 5-membered heterocyclyl, or optionally substituted 6-membered heterocyclyl.

In certain embodiments, R 2 is optionally substituted aryl, e.g., optionally substituted phenyl. In some embodiments, R 2 is optionally substituted phenyl. In some embodiments, R 2 is substituted phenyl. In some embodiments, R 2 is unsubstituted phenyl. In certain embodiments, at least one instance of R 2 is optionally substituted aryl, e.g., optionally substituted phenyl. In some embodiments, at least one instance of R 2 is optionally substituted phenyl. In some embodiments, at least one instance of R 2 is substituted phenyl. In some embodiments, at least one instance of R 2 is unsubstituted phenyl.

In certain embodiments, R 2 is optionally substituted heteroaryl, e.g., optionally substituted 5-14 membered heteroaryl, optionally substituted 5-10 membered heteroaryl, optionally substituted 5-6 membered heteroaryl, optionally substituted 5-membered heteroaryl, or optionally substituted 6-membered heteroaryl. In certain embodiments, at least one instance of R 2 is optionally substituted heteroaryl, e.g., optionally substituted 5-14 membered heteroaryl, optionally substituted 5-10 membered heteroaryl, optionally substituted 5-6 membered heteroaryl, optionally substituted 5-membered heteroaryl, or optionally substituted 6-membered heteroaryl.

In some embodiments, R 2 is a nitrogen protecting group. In some embodiments, at least one R 2 is a nitrogen protecting group.

In certain embodiments, R 2 is a group of the formula (i). In certain embodiments, R 2 is a group of the formula (ii). In certain embodiments, R 2 is a group of the formula (iii). In certain embodiments, at least one instance of R 2 is a group of the formula (i). In certain embodiments, at least one instance of R 2 is a group of the formula (ii). In certain embodiments, at least one instance of R 2 is a group of the formula (iii).

In certain embodiments, each instance of R 2 is a group other than formula (i), (ii), or (iii); in that instance, it follows that at least one R Q is a group of the formula (i), (ii), or (iii), or at least one R 1 is a group of formula (iv), and at least one of R 6 or R 7 encompassed by R 1 is a group of the formula (i), (ii), or (iii). For example, in certain embodiments, both instances of R 2 are hydrogen, and thus at least one R Q is a group of the formula (i), (ii), or (iii), or at least one R 1 is a group of formula (iv), and at least one of R 6 or R 7 encompassed by R 1 is a group of the formula (i), (ii), or (iii).

As generally defined above, each instance of R′ is independently hydrogen or optionally substituted alkyl. In some embodiments, R′ is hydrogen. In some embodiments, R′ is substituted alkyl. In certain embodiments, at least one instance of R′ is hydrogen. In certain embodiments, at least two instances of R′ is hydrogen. In certain embodiments, each instance of R′ is hydrogen. In certain embodiments, at least one instance of R′ is optionally substituted alkyl, e.g., methyl. In certain embodiments, at least two instances of R′ is optionally substituted alkyl, e.g., methyl. In some embodiments, at least one instance of R′ is hydrogen, and at least one instance of R′ is optionally substituted alkyl. In certain embodiments, one instance of R′ is optionally substituted alkyl, and the rest are hydrogen.

As generally defined above, X is O, S, or NR X . In some embodiments, X is O. In some embodiments, X is S. In some embodiments, X is NR X , wherein R X is as defined above and described herein.

As generally defined above, R X is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, or a nitrogen protecting group. In some embodiments, R X is hydrogen. In some embodiments, R X is optionally substituted alkyl. In some embodiments, R X is optionally substituted alkenyl. In some embodiments, R X is optionally substituted alkynyl. In some embodiments, R X is optionally substituted carbocyclyl. In some embodiments, R X is optionally substituted heterocyclyl. In some embodiments, R X is optionally substituted aryl. In some embodiments, R X is optionally substituted heteroaryl. In some embodiments, R X is a nitrogen protecting group.

As generally defined above, Y is O, S, or NR Y . In some embodiments, Y is O. In some embodiments, Y is S. In some embodiments, Y is NR Y , wherein R Y is as defined above and described herein.

As generally defined above, R Y is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, or a nitrogen protecting group. In some embodiments, R Y is hydrogen. In some embodiments, R Y is optionally substituted alkyl. In some embodiments, R Y is optionally substituted alkenyl. In some embodiments, R Y is optionally substituted alkynyl. In some embodiments, R Y is is optionally substituted carbocyclyl. In some embodiments, R Y is optionally substituted heterocyclyl. In some embodiments, R Y is optionally substituted aryl. In some embodiments, R Y is is optionally substituted heteroaryl. In some embodiments, R Y is a nitrogen protecting group.

As generally defined above, R P is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, an oxygen protecting group when attached to an oxygen atom, a sulfur protecting group when attached to a sulfur atom, or a nitrogen protecting group when attached to a nitrogen atom. In some embodiments, R P is hydrogen. In some embodiments, R P is optionally substituted alkyl. In some embodiments, R P is optionally substituted alkenyl. In some embodiments, R P is optionally substituted alkynyl. In some embodiments, R P is optionally substituted carbocyclyl. In some embodiments, R P is optionally substituted heterocyclyl. In some embodiments, R P is optionally substituted aryl. In some embodiments, R P is optionally substituted heteroaryl. In some embodiments, R P is an oxygen protecting group when attached to an oxygen atom. In some embodiments, R P is a sulfur protecting group when attached to a sulfur atom. In some embodiments, R P is a nitrogen protecting group when attached to a nitrogen atom.

As generally defined above, R L is optionally substituted C 1-50 alkyl, optionally substituted C 2-50 alkenyl, optionally substituted C 2-50 alkynyl, optionally substituted heteroC 1-50 alkyl, optionally substituted heteroC 2-50 alkenyl, optionally substituted heteroC 2-50 alkynyl, or a polymer.

In some embodiments, R L is optionally substituted C 1-50 alkyl. In some embodiments, R L is optionally substituted C 2-50 alkyl. In some embodiments, R L is optionally substituted C 2-40 alkyl. In some embodiments, R L is optionally substituted C 2-30 alkyl. In some embodiments, R L is optionally substituted C 2-20 alkyl. In some embodiments, R L is optionally substituted C 2-19 alkyl. In some embodiments, R L is optionally substituted C 2-18 alkyl. In some embodiments, R L is optionally substituted C 2-17 alkyl. In some embodiments, R L is optionally substituted C 2-16 alkyl. In some embodiments, R L is optionally substituted C 2-15 alkyl. In some embodiments, R L is optionally substituted C 2-14 alkyl. In some embodiments, R L is optionally substituted C 2-13 alkyl. In some embodiments, R L is optionally substituted C 2-12 alkyl. In some embodiments, R L is optionally substituted C 2-11 alkyl. In some embodiments, R L is optionally substituted C 2-10 alkyl. In some embodiments, R L is optionally substituted C 2-9 alkyl. In some embodiments, R L is optionally substituted C 2-8 alkyl. In some embodiments, R L is optionally substituted C 2-7 alkyl. In some embodiments, R L is optionally substituted C 2-6 alkyl.

In some embodiments, R L is optionally substituted C 4-50 alkyl. In some embodiments, R L is optionally substituted C 4-40 alkyl. In some embodiments, R L is optionally substituted C 4-30 alkyl. In some embodiments, R L is optionally substituted C 4-20 alkyl. In some embodiments, R L is optionally substituted C 4-19 alkyl. In some embodiments, R L is optionally substituted C 4-18 alkyl. In some embodiments, R L is optionally substituted C 4-17 alkyl. In some embodiments, R L is optionally substituted C 4-16 alkyl. In some embodiments, R L is optionally substituted C 4-15 alkyl. In some embodiments, R L is optionally substituted C 4-14 alkyl. In some embodiments, R L is optionally substituted C 4-13 alkyl. In some embodiments, R L is optionally substituted C 4-12 alkyl. In some embodiments, R L is optionally substituted C 4-11 alkyl. In some embodiments, R L is optionally substituted C 4-10 alkyl. In some embodiments, R L is optionally substituted C 4-9 alkyl. In some embodiments, R L is optionally substituted C 4-8 alkyl. In some embodiments, R L is optionally substituted C 4-7 alkyl. In some embodiments, R L is optionally substituted C 4-6 alkyl.

In some embodiments, R L is optionally substituted C 6-50 alkyl. In some embodiments, R L is optionally substituted C 6-40 alkyl. In some embodiments, R L is optionally substituted C 6-30 alkyl. In some embodiments, R L is optionally substituted C 6-20 alkyl. In some embodiments, R L is optionally substituted C 6-19 alkyl. In some embodiments, R L is optionally substituted C 6-18 alkyl. In some embodiments, R L is optionally substituted C 6-17 alkyl. In some embodiments, R L is optionally substituted C 6-16 alkyl. In some embodiments, R L is optionally substituted C 6-15 alkyl. In some embodiments, R L is optionally substituted C 6-14 alkyl. In some embodiments, R L is optionally substituted C 6-13 alkyl. In some embodiments, R L is optionally substituted C 6-12 alkyl. In some embodiments, R L is optionally substituted C 6-11 alkyl. In some embodiments, R L is optionally substituted C 6-10 alkyl. In some embodiments, R L is optionally substituted C 6-9 alkyl. In some embodiments, R L is optionally substituted C 6-8 alkyl. In some embodiments, R L is optionally substituted C 6-7 alkyl.

In some embodiments, R L is optionally substituted C 8-50 alkyl. In some embodiments, R L is optionally substituted C 8-40 alkyl. In some embodiments, R L is optionally substituted C 8-30 alkyl. In some embodiments, R L is optionally substituted C 8-20 alkyl. In some embodiments, R L is optionally substituted C 8-19 alkyl. In some embodiments, R L is optionally substituted C 8-18 alkyl. In some embodiments, R L is optionally substituted C 8-17 alkyl. In some embodiments, R L is optionally substituted C 8-16 alkyl. In some embodiments, R L is optionally substituted C 8-15 alkyl. In some embodiments, R L is optionally substituted C 8-14 alkyl. In some embodiments, R L is optionally substituted C 8-13 alkyl. In some embodiments, R L is optionally substituted C 8-12 alkyl. In some embodiments, R L is optionally substituted C 8-11 alkyl. In some embodiments, R L is optionally substituted C 8-10 alkyl. In some embodiments, R L is optionally substituted C 8-9 alkyl.

In some embodiments, R L is optionally substituted C 9-50 alkyl. In some embodiments, R L is optionally substituted C 9-40 alkyl. In some embodiments, R L is optionally substituted C 9-30 alkyl. In some embodiments, R L is optionally substituted C 9-20 alkyl. In some embodiments, R L is optionally substituted C 9-19 alkyl. In some embodiments, R L is optionally substituted C 9-18 alkyl. In some embodiments, R L is optionally substituted C 9-17 alkyl. In some embodiments, R L is optionally substituted C 9-16 alkyl. In some embodiments, R L is optionally substituted C 9-15 alkyl. In some embodiments, R L is optionally substituted C 9-14 alkyl. In some embodiments, R L is optionally substituted C 9-13 alkyl. In some embodiments, R L is optionally substituted C 9-12 alkyl. In some embodiments, R L is optionally substituted C 9-11 alkyl. In some embodiments, R L is optionally substituted C 9-10 alkyl.

In some embodiments, R L is optionally substituted C 10-50 alkyl. In some embodiments, R L is optionally substituted C 10-40 alkyl. In some embodiments, R L is optionally substituted C 10-30 alkyl. In some embodiments, R L is optionally substituted C 10-20 alkyl. In some embodiments, R L is optionally substituted C 10-19 alkyl. In some embodiments, R L is optionally substituted C 10-18 alkyl. In some embodiments, R L is optionally substituted C 10-17 alkyl. In some embodiments, R L is optionally substituted C 10-16 alkyl. In some embodiments, R L is optionally substituted C 10-15 alkyl. In some embodiments, R L is optionally substituted C 10-14 alkyl. In some embodiments, R L is optionally substituted C 10-13 alkyl. In some embodiments, R L is optionally substituted C 10-12 alkyl. In some embodiments, R L is optionally substituted C 10-11 alkyl.

In some embodiments, R L is optionally substituted C 11-50 alkyl. In some embodiments, R L is optionally substituted C 11-40 alkyl. In some embodiments, R L is optionally substituted C 11-30 alkyl. In some embodiments, R L is optionally substituted C 11-20 alkyl. In some embodiments, R L is optionally substituted C 11-19 alkyl. In some embodiments, R L is optionally substituted C 11-18 alkyl. In some embodiments, R L is optionally substituted C 11-17 alkyl. In some embodiments, R L is optionally substituted C 11-16 alkyl. In some embodiments, R L is optionally substituted C 11-15 alkyl. In some embodiments, R L is optionally substituted C 11-14 alkyl. In some embodiments, R L is optionally substituted C 11-13 alkyl. In some embodiments, R L is optionally substituted C 11-12 alkyl.

In some embodiments, R L is optionally substituted C 12-50 alkyl. In some embodiments, R L is optionally substituted C 12-40 alkyl. In some embodiments, R L is optionally substituted C 12-30 alkyl. In some embodiments, R L is optionally substituted C 12-20 alkyl. In some embodiments, R L is optionally substituted C 12-19 alkyl. In some embodiments, R L is optionally substituted C 12-18 alkyl. In some embodiments, R L is optionally substituted C 12-17 alkyl. In some embodiments, R L is optionally substituted C 12-16 alkyl. In some embodiments, R L is optionally substituted C 12-15 alkyl. In some embodiments, R L is optionally substituted C 12-14 alkyl. In some embodiments, R L is optionally substituted C 12-13 alkyl.

In some embodiments, R L is optionally substituted C 6 alkyl. In some embodiments, R L is optionally substituted C 7 alkyl. In some embodiments, R L is optionally substituted C 8 alkyl. In some embodiments, R L is optionally substituted C 9 alkyl. In some embodiments, R L is optionally substituted C 10 alkyl. In some embodiments, R L is optionally substituted C 11 alkyl. In some embodiments, R L is optionally substituted C 12 alkyl. In some embodiments, R L is optionally substituted C 13 alkyl. In some embodiments, R L is optionally substituted C 14 alkyl. In some embodiments, R L is optionally substituted C 15 alkyl. In some embodiments, R L is optionally substituted C 16 alkyl. In some embodiments, R L is optionally substituted C 17 alkyl. In some embodiments, R L is optionally substituted C 18 alkyl. In some embodiments, R L is optionally substituted C 19 alkyl. In some embodiments, R L is optionally substituted C 20 alkyl.

In some embodiments, for example, in any of the above embodiments, R L is a substituted alkyl group. In some embodiments, R L is an unsubstituted alkyl group. In some embodiments, R L is an optionally substituted straight-chain alkyl group. In some embodiments, R L is a substituted straight-chain alkyl group. In some embodiments, R L is an unsubstituted straight-chain alkyl group. In some embodiments, R L is an optionally substituted branched alkyl group. In some embodiments, R L is a substituted branched alkyl group. In some embodiments, R L is an unsubstituted branched alkyl group.

In certain embodiments, at least one instance of R L is an unsubstituted alkyl. Exemplary unsubstituted alkyl groups include, but are not limited to, —CH 3 , —C 2 H 5 , —C 3 H 7 , —C 4 H 9 , —C 5 H 11 , —C 6 H 13 , —C 7 H 15 , —C 8 H 17 , —C 9 H 19 , —C 10 H 21 , —C 11 H 23 , —C 12 H 25 , —C 13 H 27 , —C 14 H 29 , —C 15 H 31 , —C 16 H 33 , —C 17 H 35 , —C 18 H 37 , —C 19 H 39 , —C 20 H 41 , —C 21 H 43 , —C 22 H 45 , —C 23 H 47 , —C 24 H 49 , and —C 25 H 51 .

In certain embodiments, at least one instance of R L is a substituted alkyl. For example, in certain embodiments, at least one instance of R L is an alkyl substituted with one or more fluorine substituents. Exemplary fluorinated alkyl groups include, but are not limited to:

In some embodiments, R L is optionally substituted C 2-50 alkenyl. In some embodiments, R L is optionally substituted C 2-40 alkenyl. In some embodiments, R L is optionally substituted C 2-30 alkenyl. In some embodiments, R L is optionally substituted C 2-20 alkenyl. In some embodiments, R L is optionally substituted C 2-19 alkenyl. In some embodiments, R L is optionally substituted C 2-18 alkenyl. In some embodiments, R L is optionally substituted C 2-17 alkenyl. In some embodiments, R L is optionally substituted C 2-16 alkenyl. In some embodiments, R L is optionally substituted C 2-15 alkenyl. In some embodiments, R L is optionally substituted C 2-14 alkenyl. In some embodiments, R L is optionally substituted C 2-13 alkenyl. In some embodiments, R L is optionally substituted C 2-12 alkenyl. In some embodiments, R L is optionally substituted C 2-11 alkenyl. In some embodiments, R L is optionally substituted C 2-10 alkenyl. In some embodiments, R L is optionally substituted C 2-9 alkenyl. In some embodiments, R L is optionally substituted C 2-8 alkenyl. In some embodiments, R L is optionally substituted C 2-7 alkenyl. In some embodiments, R L is optionally substituted C 2-6 alkenyl.

In some embodiments, R L is optionally substituted C 4-50 alkenyl. In some embodiments, R L is optionally substituted C 4-40 alkenyl. In some embodiments, R L is optionally substituted C 4-30 alkenyl. In some embodiments, R L is optionally substituted C 4-20 alkenyl. In some embodiments, R L is optionally substituted C 4-19 alkenyl. In some embodiments, R L is optionally substituted C 4-18 alkenyl. In some embodiments, R L is optionally substituted C 4-17 alkenyl. In some embodiments, R L is optionally substituted C 4-16 alkenyl. In some embodiments, R L is optionally substituted C 4-15 alkenyl. In some embodiments, R L is optionally substituted C 4-14 alkenyl. In some embodiments, R L is optionally substituted C 4-13 alkenyl. In some embodiments, R L is optionally substituted C 4-12 alkenyl. In some embodiments, R L is optionally substituted C 4-11 alkenyl. In some embodiments, R L is optionally substituted C 4-10 alkenyl. In some embodiments, R L is optionally substituted C 4-9 alkenyl. In some embodiments, R L is optionally substituted C 4-8 alkenyl. In some embodiments, R L is optionally substituted C 4-7 alkenyl. In some embodiments, R L is optionally substituted C 4-6 alkenyl.

In some embodiments, R L is optionally substituted C 6-50 alkenyl. In some embodiments, R L is optionally substituted C 6-40 alkenyl. In some embodiments, R L is optionally substituted C 6-30 alkenyl. In some embodiments, R L is optionally substituted C 6-20 alkenyl. In some embodiments, R L is optionally substituted C 6-19 alkenyl. In some embodiments, R L is optionally substituted C 6-18 alkenyl. In some embodiments, R L is optionally substituted C 6-17 alkenyl. In some embodiments, R L is optionally substituted C 6-16 alkenyl. In some embodiments, R L is optionally substituted C 6-15 alkenyl. In some embodiments, R L is optionally substituted C 6-14 alkenyl. In some embodiments, R L is optionally substituted C 6-13 alkenyl. In some embodiments, R L is optionally substituted C 6-12 alkenyl. In some embodiments, R L is optionally substituted C 6-11 alkenyl. In some embodiments, R L is optionally substituted C 6-10 alkenyl. In some embodiments, R L is optionally substituted C 6-9 alkenyl. In some embodiments, R L is optionally substituted C 6-8 alkenyl. In some embodiments, R L is optionally substituted C 6-7 alkenyl.

In some embodiments, R L is optionally substituted C 8-50 alkenyl. In some embodiments, R L is optionally substituted C 8-40 alkenyl. In some embodiments, R L is optionally substituted C 8-30 alkenyl. In some embodiments, R L is optionally substituted C 8-20 alkenyl. In some embodiments, R L is optionally substituted C 8-19 alkenyl. In some embodiments, R L is optionally substituted C 8-18 alkenyl. In some embodiments, R L is optionally substituted C 8-17 alkenyl. In some embodiments, R L is optionally substituted C 8-16 alkenyl. In some embodiments, R L is optionally substituted C 8-15 alkenyl. In some embodiments, R L is optionally substituted C 8-14 alkenyl. In some embodiments, R L is optionally substituted C 8-13 alkenyl. In some embodiments, R L is optionally substituted C 8-12 alkenyl. In some embodiments, R L is optionally substituted C 8-11 alkenyl. In some embodiments, R L is optionally substituted C 8-10 alkenyl. In some embodiments, R L is optionally substituted C 8-9 alkenyl.

In some embodiments, R L is optionally substituted C 9-50 alkenyl. In some embodiments, R L is optionally substituted C 9-40 alkenyl. In some embodiments, R L is optionally substituted C 9-30 alkenyl. In some embodiments, R L is optionally substituted C 9-20 alkenyl. In some embodiments, R L is optionally substituted C 9-19 alkenyl. In some embodiments, R L is optionally substituted C 9-18 alkenyl. In some embodiments, R L is optionally substituted C 9-17 alkenyl. In some embodiments, R L is optionally substituted C 9-16 alkenyl. In some embodiments, R L is optionally substituted C 9-15 alkenyl. In some embodiments, R L is optionally substituted C 9-14 alkenyl. In some embodiments, R L is optionally substituted C 9-13 alkenyl. In some embodiments, R L is optionally substituted C 9-12 alkenyl. In some embodiments, R L is optionally substituted C 9-11 alkenyl. In some embodiments, R L is optionally substituted C 9-10 alkenyl.

In some embodiments, R L is optionally substituted C 10-50 alkenyl. In some embodiments, R L is optionally substituted C 10-40 alkenyl. In some embodiments, R L is optionally substituted C 10-30 alkenyl. In some embodiments, R L is optionally substituted C 10-20 alkenyl. In some embodiments, R L is optionally substituted C 10-19 alkenyl. In some embodiments, R L is optionally substituted C 10-18 alkenyl. In some embodiments, R L is optionally substituted C 10-17 alkenyl. In some embodiments, R L is optionally substituted C 10-16 alkenyl. In some embodiments, R L is optionally substituted C 1-15 alkenyl. In some embodiments, R L is optionally substituted C 10-14 alkenyl. In some embodiments, R L is optionally substituted C 10-13 alkenyl. In some embodiments, R L is optionally substituted C 10-12 alkenyl. In some embodiments, R L is optionally substituted C 10-11 alkenyl.

In some embodiments, R L is optionally substituted C 11-50 alkenyl. In some embodiments, R L is optionally substituted C 11-40 alkenyl. In some embodiments, R L is optionally substituted C 11-30 alkenyl. In some embodiments, R L is optionally substituted C 11-20 alkenyl. In some embodiments, R L is optionally substituted C 11-19 alkenyl. In some embodiments, R L is optionally substituted C 11-18 alkenyl. In some embodiments, R L is optionally substituted C 11-17 alkenyl. In some embodiments, R L is optionally substituted C 11-16 alkenyl. In some embodiments, R L is optionally substituted C 11-15 alkenyl. In some embodiments, R L is optionally substituted C 11-14 alkenyl. In some embodiments, R L is optionally substituted C 11-13 alkenyl. In some embodiments, R L is optionally substituted C 11-12 alkenyl.

In some embodiments, R L is optionally substituted C 12-50 alkenyl. In some embodiments, R L is optionally substituted C 12-40 alkenyl. In some embodiments, R L is optionally substituted C 12-30 alkenyl. In some embodiments, R L is optionally substituted C 12-20 alkenyl. In some embodiments, R L is optionally substituted C 12-19 alkenyl. In some embodiments, R L is optionally substituted C 12-18 alkenyl. In some embodiments, R L is optionally substituted C 12-17 alkenyl. In some embodiments, R L is optionally substituted C 12-16 alkenyl. In some embodiments, R L is optionally substituted C 12-15 alkenyl. In some embodiments, R L is optionally substituted C 12-14 alkenyl. In some embodiments, R L is optionally substituted C 12-13 alkenyl.

In some embodiments, R L is optionally substituted C 6 alkenyl. In some embodiments, R L is optionally substituted C 7 alkenyl. In some embodiments, R L is optionally substituted C 8 alkenyl. In some embodiments, R L is optionally substituted C 9 alkenyl. In some embodiments, R L is optionally substituted C 10 alkenyl. In some embodiments, R L is optionally substituted C11 alkenyl. In some embodiments, R L is optionally substituted C 12 alkenyl. In some embodiments, R L is optionally substituted C 13 alkenyl. In some embodiments, R L is optionally substituted C 14 alkenyl. In some embodiments, R L is optionally substituted C 15 alkenyl. In some embodiments, R L is optionally substituted C 16 alkenyl. In some embodiments, R L is optionally substituted C 17 alkenyl. In some embodiments, R L is optionally substituted C 18 alkenyl. In some embodiments, R L is optionally substituted C 19 alkenyl. In some embodiments, R L is optionally substituted C 20 alkenyl.

In some embodiments, for example, in any of the above embodiments, R L is a substituted alkyl group. In some embodiments, R L is an unsubstituted alkyl group. In some embodiments, R L is an optionally substituted straight-chain alkenyl group. In some embodiments, R L is a substituted straight-chain alkenyl group. In some embodiments, R L is an unsubstituted straight-chain alkenyl group. In some embodiments, R L is an optionally substituted branched alkenyl group. In some embodiments, R L is a substituted branched alkenyl group. In some embodiments, R L is an unsubstituted branched alkenyl group.

Exemplary unsubstituted alkenyl group include, but are not limited to:

Myristoleic —(C H2 ) 7 CH═CH(CH 2 ) 3 CH 3 ,

Palmitoliec —(CH) 7 CH═CH(CH 2 ) 5 CH 3 ,

Sapienic —(CH 2 ) 4 CH═CH(CH 2 ) 8 CH 3 ,

Oleic —(CH 2 ) 7 CH═CH(CH 2 ) 7 CH 3 ,

Linoleic —(CH 2 ) 7 CH═CHCH 2 CH═CH(CH 2 ) 4 CH 3 ,

α-linolenic —(CH 2 ) 7 CH═CHCH 2 CH═CHCH 2 CH═CHCH 2 CH═CH(CH 2 ) 4 CH 3 ,

Arachinodonic —(CH 2 ) 3 CH═CHCH 2 CH═CHCH 2 CH═CHCH 2 CH═CH(CH 2 ) 4 CH 3 ,

Eicosapentaenoic —(CH 2 ) 3 CH═CHCH 2 CH═CHCH 2 CH═CHCH 2 CH═CHCH 2 CH═CHCH 2 CH 3 ,

Erucic —(CH 2 ) 3 CH═CH(CH 2 ) 7 CH 3 , and

Docosahexaenoic —(CH 2 ) 2 CH═CHCH 2 CH═CHCH 2 CH═CHCH 2 CH═CHCH 2 CH═CHCH 2 CH═CH—CH 2 CH 3 .

In some embodiments, wherein R L is defined as a C 6-50 alkyl or C 6-50 alkenyl groups, such groups are meant to encompass lipophilic groups (also referred to as a “lipid tail”). Lipophilic groups comprise a group of molecules that include fats, waxes, oils, fatty acids, and the like. Lipid tails present in these lipid groups can be saturated and unsaturated, depending on whether or not the lipid tail comprises double bonds. The lipid tail can also comprise different lengths, often categorized as medium (i.e., with tails between 7-12 carbons, e.g., C 7-12 alkyl or C 7-12 alkenyl), long (i.e., with tails greater than 12 carbons and up to 22 carbons, e.g., C 13-22 alkyl or C 13-22 alkenyl), or very long (i.e., with tails greater than 22 carbons, e.g., C 23-30 alkyl or C 23-30 alkenyl).

In some embodiments, R L is optionally substituted C 2-50 alkynyl. In some embodiments, R L is optionally substituted C 2-40 alkynyl. In some embodiments, R L is optionally substituted C 2-30 alkynyl. In some embodiments, R L is optionally substituted C 2-20 alkynyl. In some embodiments, R L is optionally substituted C 2-19 alkynyl. In some embodiments, R L is optionally substituted C 2-18 alkynyl. In some embodiments, R L is optionally substituted C 2-17 alkynyl. In some embodiments, R L is optionally substituted C 2-16 alkynyl. In some embodiments, R L is optionally substituted C 2-15 alkynyl. In some embodiments, R L is optionally substituted C 2-14 alkynyl. In some embodiments, R L is optionally substituted C 2-13 alkynyl. In some embodiments, R L is optionally substituted C 2-12 alkynyl. In some embodiments, R L is optionally substituted C 2-11 alkynyl. In some embodiments, R L is optionally substituted C 2-10 alkynyl. In some embodiments, R L is optionally substituted C 2-9 alkynyl. In some embodiments, R L is optionally substituted C 2-8 alkynyl. In some embodiments, R L is optionally substituted C 2-7 alkynyl. In some embodiments, R L is optionally substituted C 2-6 alkynyl.

In some embodiments, R L is optionally substituted C 4-50 alkynyl. In some embodiments, R L is optionally substituted C 4-40 alkynyl. In some embodiments, R L is optionally substituted C 4-30 alkynyl. In some embodiments, R L is optionally substituted C 4-20 alkynyl. In some embodiments, R L is optionally substituted C 4-19 alkynyl. In some embodiments, R L is optionally substituted C 4-18 alkynyl. In some embodiments, R L is optionally substituted C 4-17 alkynyl. In some embodiments, R L is optionally substituted C 4-16 alkynyl. In some embodiments, R L is optionally substituted C 4-15 alkynyl. In some embodiments, R L is optionally substituted C 4-14 alkynyl. In some embodiments, R L is optionally substituted C 4-13 alkynyl. In some embodiments, R L is optionally substituted C 4-12 alkynyl. In some embodiments, R L is optionally substituted C 4-11 alkynyl. In some embodiments, R L is optionally substituted C 4-10 alkynyl. In some embodiments, R L is optionally substituted C 4-9 alkynyl. In some embodiments, R L is optionally substituted C 4-8 alkynyl. In some embodiments, R L is optionally substituted C 4-7 alkynyl. In some embodiments, R L is optionally substituted C 4-6 alkynyl.

In some embodiments, R L is optionally substituted C 6-50 alkynyl. In some embodiments, R L is optionally substituted C 6-40 alkynyl. In some embodiments, R L is optionally substituted C 6-30 alkynyl. In some embodiments, R L is optionally substituted C 6-20 alkynyl. In some embodiments, R L is optionally substituted C 6-19 alkynyl. In some embodiments, R L is optionally substituted C 6-18 alkynyl. In some embodiments, R L is optionally substituted C 6-17 alkynyl. In some embodiments, R L is optionally substituted C 6-16 alkynyl. In some embodiments, R L is optionally substituted C 6-15 alkynyl. In some embodiments, R L is optionally substituted C 6-14 alkynyl. In some embodiments, R L is optionally substituted C 6-13 alkynyl. In some embodiments, R L is optionally substituted C 6-12 alkynyl. In some embodiments, R L is optionally substituted C 6-11 alkynyl. In some embodiments, R L is optionally substituted C 6-10 alkynyl. In some embodiments, R L is optionally substituted C 6-9 alkynyl. In some embodiments, R L is optionally substituted C 6-8 alkynyl. In some embodiments, R L is optionally substituted C 6-7 alkynyl.

In some embodiments, R L is optionally substituted C 8-50 alkynyl. In some embodiments, R L is optionally substituted C 8-40 alkynyl. In some embodiments, R L is optionally substituted C 8-30 alkynyl. In some embodiments, R L is optionally substituted C 8-20 alkynyl. In some embodiments, R L is optionally substituted C 8-19 alkynyl. In some embodiments, R L is optionally substituted C 8-18 alkynyl. In some embodiments, R L is optionally substituted C 8-17 alkynyl. In some embodiments, R L is optionally substituted C 8-16 alkynyl. In some embodiments, R L is optionally substituted C 8-15 alkynyl. In some embodiments, R L is optionally substituted C 8-14 alkynyl. In some embodiments, R L is optionally substituted C 8-13 alkynyl. In some embodiments, R L is optionally substituted C 8-12 alkynyl. In some embodiments, R L is optionally substituted C 8-11 alkynyl. In some embodiments, R L is optionally substituted C 8-10 alkynyl. In some embodiments, R L is optionally substituted C 8-9 alkynyl.

In some embodiments, R L is optionally substituted C 9-50 alkynyl. In some embodiments, R L is optionally substituted C 9-40 alkynyl. In some embodiments, R L is optionally substituted C 9-30 alkynyl. In some embodiments, R L is optionally substituted C 9-20 alkynyl. In some embodiments, R L is optionally substituted C 9-19 alkynyl. In some embodiments, R L is optionally substituted C 9-18 alkynyl. In some embodiments, R L is optionally substituted C 9-17 alkynyl. In some embodiments, R L is optionally substituted C 9-16 alkynyl. In some embodiments, R L is optionally substituted C 9-15 alkynyl. In some embodiments, R L is optionally substituted C 9-14 alkynyl. In some embodiments, R L is optionally substituted C 9-13 alkynyl. In some embodiments, R L is optionally substituted C 9-12 alkynyl. In some embodiments, R L is optionally substituted C 9-11 alkynyl. In some embodiments, R L is optionally substituted C 9-10 alkynyl.

In some embodiments, R L is optionally substituted C 10-50 alkynyl. In some embodiments, R L is optionally substituted C 10-40 alkynyl. In some embodiments, R L is optionally substituted C 10-30 alkynyl. In some embodiments, R L is optionally substituted C 10-20 alkynyl. In some embodiments, R L is optionally substituted C 10-19 alkynyl. In some embodiments, R L is optionally substituted C 10-18 alkynyl. In some embodiments, R L is optionally substituted C 10-17 alkynyl. In some embodiments, R L is optionally substituted C 10-16 alkynyl. In some embodiments, R L is optionally substituted C 10-15 alkynyl. In some embodiments, R L is optionally substituted C 10-14 alkynyl. In some embodiments, R L is optionally substituted C 10-13 alkynyl. In some embodiments, R L is optionally substituted C 10-12 alkynyl. In some embodiments, R L is optionally substituted C 10-11 alkynyl.

In some embodiments, R L is optionally substituted C 11-50 alkynyl. In some embodiments, R L is optionally substituted C 11-40 alkynyl. In some embodiments, R L is optionally substituted C 11-30 alkynyl. In some embodiments, R L is optionally substituted C 11-20 alkynyl. In some embodiments, R L is optionally substituted C 11-19 alkynyl. In some embodiments, R L is optionally substituted C 11-18 alkynyl. In some embodiments, R L is optionally substituted C 11-17 alkynyl. In some embodiments, R L is optionally substituted C 11-16 alkynyl. In some embodiments, R L is optionally substituted C 11-15 alkynyl. In some embodiments, R L is optionally substituted C 11-14 alkynyl. In some embodiments, R L is optionally substituted C 11-13 alkynyl. In some embodiments, R L is optionally substituted C 11-12 alkynyl.

In some embodiments, R L is optionally substituted C 12-50 alkynyl. In some embodiments, R L is optionally substituted C 12-40 alkynyl. In some embodiments, R L is optionally substituted C 12-30 alkynyl. In some embodiments, R L is optionally substituted C 12-20 alkynyl. In some embodiments, R L is optionally substituted C 12-19 alkynyl. In some embodiments, R L is optionally substituted C 12-18 alkynyl. In some embodiments, R L is optionally substituted C 12-17 alkynyl. In some embodiments, R L is optionally substituted C 12-16 alkynyl. In some embodiments, R L is optionally substituted C 12-15 alkynyl. In some embodiments, R L is optionally substituted C 12-14 alkynyl. In some embodiments, R L is optionally substituted C 12-13 alkynyl.

In some embodiments, R L is optionally substituted C 6 alkynyl. In some embodiments, R L is optionally substituted C 7 alkynyl. In some embodiments, R L is optionally substituted C 8 alkynyl. In some embodiments, R L is optionally substituted C 9 alkynyl. In some embodiments, R L is optionally substituted C 10 alkynyl. In some embodiments, R L is optionally substituted C 11 alkynyl. In some embodiments, R L is optionally substituted C 12 alkynyl. In some embodiments, R L is optionally substituted C 13 alkynyl. In some embodiments, R L is optionally substituted C 14 alkynyl. In some embodiments, R L is optionally substituted C 15 alkynyl. In some embodiments, R L is optionally substituted C 16 alkynyl. In some embodiments, R L is optionally substituted C 17 alkynyl. In some embodiments, R L is optionally substituted C 18 alkynyl. In some embodiments, R L is optionally substituted C 19 alkynyl. In some embodiments, R L is optionally substituted C 20 alkynyl.

In some embodiments, for example, in any of the above embodiments, R L is a substituted alkynyl group. In some embodiments, R L is an unsubstituted alkynyl group. In some embodiments, R L is an optionally substituted straight-chain alkyl group. In some embodiments, R L is an optionally substituted straight-chain alkynyl group. In some embodiments, R L is a substituted straight-chain alkynyl group. In some embodiments, R L is an unsubstituted straight-chain alkynyl group. In some embodiments, R L is an optionally substituted branched alkynyl group. In some embodiments, R L is a substituted branched alkynyl group. In some embodiments, R L is an unsubstituted branched alkynyl group.

In some embodiments, R L is optionally substituted heteroC 1-50 alkyl. In some embodiments, R L is optionally substituted heteroC 2-50 alkyl. In some embodiments, R L is optionally substituted heteroC 2-40 alkyl. In some embodiments, R L is optionally substituted heteroC 2-30 alkyl. In some embodiments, R L is optionally substituted heteroC 2-20 alkyl. In some embodiments, R L is optionally substituted heteroC 2-19 alkyl. In some embodiments, R L is optionally substituted heteroC 2-18 alkyl. In some embodiments, R L is optionally substituted heteroC 2-17 alkyl. In some embodiments, R L is optionally substituted heteroC 2-16 alkyl. In some embodiments, R L is optionally substituted heteroC 2-15 alkyl. In some embodiments, R L is optionally substituted heteroC 2-14 alkyl. In some embodiments, R L is optionally substituted heteroC 2-13 alkyl. In some embodiments, R L is optionally substituted heteroC 2-12 alkyl. In some embodiments, R L is optionally substituted heteroC 2-11 alkyl. In some embodiments, R L is optionally substituted heteroC 2-10 alkyl. In some embodiments, R L is optionally substituted heteroC 2-9 alkyl. In some embodiments, R L is optionally substituted heteroC 2-8 alkyl. In some embodiments, R L is optionally substituted heteroC 2-7 alkyl. In some embodiments, R L is optionally substituted heteroC 2-6 alkyl.

In some embodiments, R L is optionally substituted heteroC 4-50 alkyl. In some embodiments, R L is optionally substituted heteroC 4-40 alkyl. In some embodiments, R L is optionally substituted heteroC 4-30 alkyl. In some embodiments, R L is optionally substituted heteroC 4-20 alkyl. In some embodiments, R L is optionally substituted heteroC 4-19 alkyl. In some embodiments, R L is optionally substituted heteroC 4-18 alkyl. In some embodiments, R L is optionally substituted heteroC 4-17 alkyl. In some embodiments, R L is optionally substituted heteroC 4-16 alkyl. In some embodiments, R L is optionally substituted heteroC 4-15 alkyl. In some embodiments, R L is optionally substituted heteroC 4-14 alkyl. In some embodiments, R L is optionally substituted heteroC 4-13 alkyl. In some embodiments, R L is optionally substituted heteroC 4-12 alkyl. In some embodiments, R L is optionally substituted heteroC 4-11 alkyl. In some embodiments, R L is optionally substituted heteroC 4-10 alkyl. In some embodiments, R L is optionally substituted heteroC 4-9 alkyl. In some embodiments, R L is optionally substituted heteroC 4-8 alkyl. In some embodiments, R L is optionally substituted heteroC 4-7 alkyl. In some embodiments, R L is optionally substituted heteroC 4-6 alkyl.

In some embodiments, R L is optionally substituted heteroC 6-50 alkyl. In some embodiments, R L is optionally substituted heteroC 6-40 alkyl. In some embodiments, R L is optionally substituted heteroC 6-30 alkyl. In some embodiments, R L is optionally substituted heteroC 6-20 alkyl. In some embodiments, R L is optionally substituted heteroC 6-19 alkyl. In some embodiments, R L is optionally substituted heteroC 6-18 alkyl. In some embodiments, R L is optionally substituted heteroC 6-17 alkyl. In some embodiments, R L is optionally substituted heteroC 6-16 alkyl. In some embodiments, R L is optionally substituted heteroC 6-15 alkyl. In some embodiments, R L is optionally substituted heteroC 6-14 alkyl. In some embodiments, R L is optionally substituted heteroC 6-13 alkyl. In some embodiments, R L is optionally substituted heteroC 6-12 alkyl. In some embodiments, R L is optionally substituted heteroC 6-11 alkyl. In some embodiments, R L is optionally substituted heteroC 6-10 alkyl. In some embodiments, R L is optionally substituted heteroC 6-9 alkyl. In some embodiments, R L is optionally substituted heteroC 6-8 alkyl. In some embodiments, R L is optionally substituted heteroC 6-7 alkyl.

In some embodiments, R L is optionally substituted heteroC 8-50 alkyl. In some embodiments, R L is optionally substituted heteroC 8-40 alkyl. In some embodiments, R L is optionally substituted heteroC 8-30 alkyl. In some embodiments, R L is optionally substituted heteroC 8-20 alkyl. In some embodiments, R L is optionally substituted heteroC 8-19 alkyl. In some embodiments, R L is optionally substituted heteroC 8-18 alkyl. In some embodiments, R L is optionally substituted heteroC 8-17 alkyl. In some embodiments, R L is optionally substituted heteroC 8-16 alkyl. In some embodiments, R L is optionally substituted heteroC 8-15 alkyl. In some embodiments, R L is optionally substituted heteroC 8-14 alkyl. In some embodiments, R L is optionally substituted heteroC 8-13 alkyl. In some embodiments, R L is optionally substituted heteroC 8-12 alkyl. In some embodiments, R L is optionally substituted heteroC 8-11 alkyl. In some embodiments, R L is optionally substituted heteroC 8-10 alkyl. In some embodiments, R L is optionally substituted heteroC 8-9 alkyl.

In some embodiments, R L is optionally substituted heteroC 9-50 alkyl. In some embodiments, R L is optionally substituted heteroC 9-40 alkyl. In some embodiments, R L is optionally substituted heteroC 9-30 alkyl. In some embodiments, R L is optionally substituted heteroC 9-20 alkyl. In some embodiments, R L is optionally substituted heteroC 9-19 alkyl. In some embodiments, R L is optionally substituted heteroC 9-18 alkyl. In some embodiments, R L is optionally substituted heteroC 9-17 alkyl. In some embodiments, R L is optionally substituted heteroC 9-16 alkyl. In some embodiments, R L is optionally substituted heteroC 9-15 alkyl. In some embodiments, R L is optionally substituted heteroC 9-14 alkyl. In some embodiments, R L is optionally substituted heteroC 9-13 alkyl. In some embodiments, R L is optionally substituted heteroC 9-12 alkyl. In some embodiments, R L is optionally substituted heteroC 9-11 alkyl. In some embodiments, R L is optionally substituted heteroC 9-10 alkyl.

In some embodiments, R L is optionally substituted heteroC 10-50 alkyl. In some embodiments, R L is optionally substituted heteroC 10-40 alkyl. In some embodiments, R L is optionally substituted heteroC 10-30 alkyl. In some embodiments, R L is optionally substituted heteroC 10-20 alkyl. In some embodiments, R L is optionally substituted heteroC 10-19 alkyl. In some embodiments, R L is optionally substituted heteroC 10-18 alkyl. In some embodiments, R L is optionally substituted heteroC 10-17 alkyl. In some embodiments, R L is optionally substituted heteroC 10-16 alkyl. In some embodiments, R L is optionally substituted heteroC 10-15 alkyl. In some embodiments, R L is optionally substituted heteroC 10-14 alkyl. In some embodiments, R L is optionally substituted heteroC 10-13 alkyl. In some embodiments, R L is optionally substituted heteroC 10-12 alkyl. In some embodiments, R L is optionally substituted heteroC 10-11 alkyl.

In some embodiments, R L is optionally substituted heteroC 11-50 alkyl. In some embodiments, R L is optionally substituted heteroC 11-40 alkyl. In some embodiments, R L is optionally substituted heteroC 11-30 alkyl. In some embodiments, R L is optionally substituted heteroC 11-20 alkyl. In some embodiments, R L is optionally substituted heteroC 11-19 alkyl. In some embodiments, R L is optionally substituted heteroC 11-18 alkyl. In some embodiments, R L is optionally substituted heteroC 11-17 alkyl. In some embodiments, R L is optionally substituted heteroC 11-16 alkyl. In some embodiments, R L is optionally substituted heteroC 11-15 alkyl. In some embodiments, R L is optionally substituted heteroC 11-14 alkyl. In some embodiments, R L is optionally substituted heteroC 11-13 alkyl. In some embodiments, R L is optionally substituted heteroC 11-12 alkyl.

In some embodiments, R L is optionally substituted heteroC 12-50 alkyl. In some embodiments, R L is optionally substituted heteroC 12-40 alkyl. In some embodiments, R L is optionally substituted heteroC 12-30 alkyl. In some embodiments, R L is optionally substituted heteroC 12-20 alkyl. In some embodiments, R L is optionally substituted heteroC 12-19 alkyl. In some embodiments, R L is optionally substituted heteroC 12-18 alkyl. In some embodiments, R L is optionally substituted heteroC 12-17 alkyl. In some embodiments, R L is optionally substituted heteroC 12-16 alkyl. In some embodiments, R L is optionally substituted heteroC 12-15 alkyl. In some embodiments, R L is optionally substituted heteroC 12-14 alkyl. In some embodiments, R L is optionally substituted heteroC 12-13 alkyl.

In some embodiments, R L is optionally substituted heteroC 6 alkyl. In some embodiments, R L is optionally substituted heteroC 7 alkyl. In some embodiments, R L is optionally substituted heteroC 8 alkyl. In some embodiments, R L is optionally substituted heteroC 9 alkyl. In some embodiments, R L is optionally substituted heteroC 10 alkyl. In some embodiments, R L is optionally substituted heteroC 11 alkyl. In some embodiments, R L is optionally substituted heteroC 12 alkyl. In some embodiments, R L is optionally substituted heteroC 13 alkyl. In some embodiments, R L is optionally substituted heteroC 14 alkyl. In some embodiments, R L is optionally substituted heteroC 15 alkyl. In some embodiments, R L is optionally substituted heteroC 16 alkyl. In some embodiments, R L is optionally substituted heteroC 17 alkyl. In some embodiments, R L is optionally substituted heteroC 18 alkyl. In some embodiments, R L is optionally substituted heteroC 19 alkyl. In some embodiments, R L is optionally substituted heteroC 20 alkyl.

In some embodiments, for example, in any of the above embodiments, R L is a substituted heteroalkyl group. In some embodiments, R L is an unsubstituted heteroalkyl group. In some embodiments, R L is an optionally substituted straight-chain heteroalkyl group. In some embodiments, R L is a substituted straight-chain heteroalkyl group. In some embodiments, R L is an unsubstituted straight-chain heteroalkyl group. In some embodiments, R L is an optionally substituted branched heteroalkyl group. In some embodiments, R L is a substituted branched heteroalkyl group. In some embodiments, R L is an unsubstituted branched heteroalkyl group.

Exemplary unsubstituted heteroalkyl groups include, but are not limited to:

In some embodiments, R L is optionally substituted heteroC 2-50 alkenyl. In some embodiments, R L is optionally substituted heteroC 2-40 alkenyl. In some embodiments, R L is optionally substituted heteroC 2-30 alkenyl. In some embodiments, R L is optionally substituted heteroC 2-20 alkenyl. In some embodiments, R L is optionally substituted heteroC 2-19 alkenyl. In some embodiments, R L is optionally substituted heteroC 2-18 alkenyl. In some embodiments, R L is optionally substituted heteroC 2-17 alkenyl. In some embodiments, R L is optionally substituted heteroC 2-16 alkenyl. In some embodiments, R L is optionally substituted heteroC 2-15 alkenyl. In some embodiments, R L is optionally substituted heteroC 2-14 alkenyl. In some embodiments, R L is optionally substituted heteroC 2-13 alkenyl. In some embodiments, R L is optionally substituted heteroC 2-12 alkenyl. In some embodiments, R L is optionally substituted heteroC 2-11 alkenyl. In some embodiments, R L is optionally substituted heteroC 2-10 alkenyl. In some embodiments, R L is optionally substituted heteroC 2-9 alkenyl. In some embodiments, R L is optionally substituted heteroC 2-8 alkenyl. In some embodiments, R L is optionally substituted heteroC 2-7 alkenyl. In some embodiments, R L is optionally substituted heteroC 2-6 alkenyl.

In some embodiments, R L is optionally substituted heteroC 4-50 alkenyl. In some embodiments, R L is optionally substituted heteroC 4-40 alkenyl. In some embodiments, R L is optionally substituted heteroC 4-30 alkenyl. In some embodiments, R L is optionally substituted heteroC 4-20 alkenyl. In some embodiments, R L is optionally substituted heteroC 4-19 alkenyl. In some embodiments, R L is optionally substituted heteroC 4-18 alkenyl. In some embodiments, R L is optionally substituted heteroC 4-17 alkenyl. In some embodiments, R L is optionally substituted heteroC 4-16 alkenyl. In some embodiments, R L is optionally substituted heteroC 4-15 alkenyl. In some embodiments, R L is optionally substituted heteroC 4-14 alkenyl. In some embodiments, R L is optionally substituted heteroC 4-13 alkenyl. In some embodiments, R L is optionally substituted heteroC 4-12 alkenyl. In some embodiments, R L is optionally substituted heteroC 4-11 alkenyl. In some embodiments, R L is optionally substituted heteroC 4-10 alkenyl. In some embodiments, R L is optionally substituted heteroC 4-9 alkenyl. In some embodiments, R L is optionally substituted heteroC 4-8 alkenyl. In some embodiments, R L is optionally substituted heteroC 4-7 alkenyl. In some embodiments, R L is optionally substituted heteroC 4-6 alkenyl.

In some embodiments, R L is optionally substituted heteroC 6-50 alkenyl. In some embodiments, R L is optionally substituted heteroC 6-40 alkenyl. In some embodiments, R L is optionally substituted heteroC 6-30 alkenyl. In some embodiments, R L is optionally substituted heteroC 6-20 alkenyl. In some embodiments, R L is optionally substituted heteroC 6-19 alkenyl. In some embodiments, R L is optionally substituted heteroC 6-18 alkenyl. In some embodiments, R L is optionally substituted heteroC 6-17 alkenyl. In some embodiments, R L is optionally substituted heteroC 6-16 alkenyl. In some embodiments, R L is optionally substituted heteroC 6-15 alkenyl. In some embodiments, R L is optionally substituted heteroC 6-14 alkenyl. In some embodiments, R L is optionally substituted heteroC 6-13 alkenyl. In some embodiments, R L is optionally substituted heteroC 6-12 alkenyl. In some embodiments, R L is optionally substituted heteroC 6-11 alkenyl. In some embodiments, R L is optionally substituted heteroC 6-10 alkenyl. In some embodiments, R L is optionally substituted heteroC 6-9 alkenyl. In some embodiments, R L is optionally substituted heteroC 6-8 alkenyl. In some embodiments, R L is optionally substituted heteroC 6-7 alkenyl.

In some embodiments, R L is optionally substituted heteroC 8-50 alkenyl. In some embodiments, R L is optionally substituted heteroC 8-40 alkenyl. In some embodiments, R L is optionally substituted heteroC 8-30 alkenyl. In some embodiments, R L is optionally substituted heteroC 8-20 alkenyl. In some embodiments, R L is optionally substituted heteroC 8-19 alkenyl. In some embodiments, R L is optionally substituted heteroC 8-18 alkenyl. In some embodiments, R L is optionally substituted heteroC 8-17 alkenyl. In some embodiments, R L is optionally substituted heteroC 8-16 alkenyl. In some embodiments, R L is optionally substituted heteroC 8-15 alkenyl. In some embodiments, R L is optionally substituted heteroC 8-14 alkenyl. In some embodiments, R L is optionally substituted heteroC 8-13 alkenyl. In some embodiments, R L is optionally substituted heteroC 8-12 alkenyl. In some embodiments, R L is optionally substituted heteroC 8-11 alkenyl. In some embodiments, R L is optionally substituted heteroC 8-10 alkenyl. In some embodiments, R L is optionally substituted heteroC 8-9 alkenyl.

In some embodiments, R L is optionally substituted heteroC 9-50 alkenyl. In some embodiments, R L is optionally substituted heteroC 9-40 alkenyl. In some embodiments, R L is optionally substituted heteroC 9-30 alkenyl. In some embodiments, R L is optionally substituted heteroC 9-20 alkenyl. In some embodiments, R L is optionally substituted heteroC 9-19 alkenyl. In some embodiments, R L is optionally substituted heteroC 9-18 alkenyl. In some embodiments, R L is optionally substituted heteroC 9-17 alkenyl. In some embodiments, R L is optionally substituted heteroC 9-16 alkenyl. In some embodiments, R L is optionally substituted heteroC 9-15 alkenyl. In some embodiments, R L is optionally substituted heteroC 9-14 alkenyl. In some embodiments, R L is optionally substituted heteroC 9-13 alkenyl. In some embodiments, R L is optionally substituted heteroC 9-12 alkenyl. In some embodiments, R L is optionally substituted heteroC 9-11 alkenyl. In some embodiments, R L is optionally substituted heteroC 9-10 alkenyl.

In some embodiments, R L is optionally substituted heteroC 10-50 alkenyl. In some embodiments, R L is optionally substituted heteroC 10-40 alkenyl. In some embodiments, R L is optionally substituted heteroC 10-30 alkenyl. In some embodiments, R L is optionally substituted heteroC 10-20 alkenyl. In some embodiments, R L is optionally substituted heteroC 10-19 alkenyl. In some embodiments, R L is optionally substituted heteroC 10-18 alkenyl. In some embodiments, R L is optionally substituted heteroC 10-17 alkenyl. In some embodiments, R L is optionally substituted heteroC 10-16 alkenyl. In some embodiments, R L is optionally substituted heteroC 11-15 alkenyl. In some embodiments, R L is optionally substituted heteroC 10-14 alkenyl. In some embodiments, R L is optionally substituted heteroC 10-13 alkenyl. In some embodiments, R L is optionally substituted heteroC 10-12 alkenyl. In some embodiments, R L is optionally substituted heteroC 10-11 alkenyl.

In some embodiments, R L is optionally substituted heteroC 11-50 alkenyl. In some embodiments, R L is optionally substituted heteroC 11-40 alkenyl. In some embodiments, R L is optionally substituted heteroC 11-30 alkenyl. In some embodiments, R L is optionally substituted heteroC 11-20 alkenyl. In some embodiments, R L is optionally substituted heteroC 11-19 alkenyl. In some embodiments, R L is optionally substituted heteroC 11-18 alkenyl. In some embodiments, R L is optionally substituted heteroC 11-17 alkenyl. In some embodiments, R L is optionally substituted heteroC 11-16 alkenyl. In some embodiments, R L is optionally substituted heteroC 11-15 alkenyl. In some embodiments, R L is optionally substituted heteroC 11-14 alkenyl. In some embodiments, R L is optionally substituted heteroC 11-13 alkenyl. In some embodiments, R L is optionally substituted heteroC 11-12 alkenyl.

In some embodiments, R L is optionally substituted heteroC 12-50 alkenyl. In some embodiments, R L is optionally substituted heteroC 12-40 alkenyl. In some embodiments, R L is optionally substituted heteroC 12-30 alkenyl. In some embodiments, R L is optionally substituted heteroC 12-20 alkenyl. In some embodiments, R L is optionally substituted heteroC 12-19 alkenyl. In some embodiments, R L is optionally substituted heteroC 12-18 alkenyl. In some embodiments, R L is optionally substituted heteroC 12-17 alkenyl. In some embodiments, R L is optionally substituted heteroC 12-16 alkenyl. In some embodiments, R L is optionally substituted heteroC 12-15 alkenyl. In some embodiments, R L is optionally substituted heteroC 12-14 alkenyl. In some embodiments, R L is optionally substituted heteroC 12-13 alkenyl.

In some embodiments, R L is optionally substituted heteroC 6 alkenyl. In some embodiments, R L is optionally substituted heteroC 7 alkenyl. In some embodiments, R L is optionally substituted heteroC 8 alkenyl. In some embodiments, R L is optionally substituted heteroC 9 alkenyl. In some embodiments, R L is optionally substituted heteroC 10 alkenyl. In some embodiments, R L is optionally substituted heteroC 11 alkenyl. In some embodiments, R L is optionally substituted heteroC 12 alkenyl. In some embodiments, R L is optionally substituted heteroC 13 alkenyl. In some embodiments, R L is optionally substituted heteroC 14 alkenyl. In some embodiments, R L is optionally substituted heteroC 15 alkenyl. In some embodiments, R L is optionally substituted heteroC 16 alkenyl. In some embodiments, R L is optionally substituted heteroC 17 alkenyl. In some embodiments, R L is optionally substituted heteroC 18 alkenyl. In some embodiments, R L is optionally substituted heteroC 19 alkenyl. In some embodiments, R L is optionally substituted heteroC 20 alkenyl.

In some embodiments, for example, in any of the above embodiments, R L is a substituted heteroalkenyl group. In some embodiments, R L is an unsubstituted heteroalkenyl group. In some embodiments, R L is an optionally substituted straight-chain heteroalkenyl group. In some embodiments, R L is a substituted straight-chain heteroalkenyl group. In some embodiments, R L is an unsubstituted straight-chain heteroalkenyl group. In some embodiments, R L is an optionally substituted branched heteroalkenyl group. In some embodiments, R L is a substituted branched heteroalkenyl group. In some embodiments, R L is an unsubstituted branched heteroalkenyl group.

In some embodiments, R L is optionally substituted heteroC 2-50 alkynyl. In some embodiments, R L is optionally substituted heteroC 2-40 alkynyl. In some embodiments, R L is optionally substituted heteroC 2-30 alkynyl. In some embodiments, R L is optionally substituted heteroC 2-20 alkynyl. In some embodiments, R L is optionally substituted heteroC 2-19 alkynyl. In some embodiments, R L is optionally substituted heteroC 2-18 alkynyl. In some embodiments, R L is optionally substituted heteroC 2-17 alkynyl. In some embodiments, R L is optionally substituted heteroC 2-16 alkynyl. In some embodiments, R L is optionally substituted heteroC 2-15 alkynyl. In some embodiments, R L is optionally substituted heteroC 2-14 alkynyl. In some embodiments, R L is optionally substituted heteroC 2-13 alkynyl. In some embodiments, R L is optionally substituted heteroC 2-12 alkynyl. In some embodiments, R L is optionally substituted heteroC 2-11 alkynyl. In some embodiments, R L is optionally substituted heteroC 2-10 alkynyl. In some embodiments, R L is optionally substituted heteroC 2-9 alkynyl. In some embodiments, R L is optionally substituted heteroC 2-8 alkynyl. In some embodiments, R L is optionally substituted heteroC 2-7 alkynyl. In some embodiments, R L is optionally substituted heteroC 2-6 alkynyl.

In some embodiments, R L is optionally substituted heteroC 4-50 alkynyl. In some embodiments, R L is optionally substituted heteroC 4-40 alkynyl. In some embodiments, R L is optionally substituted heteroC 4-30 alkynyl. In some embodiments, R L is optionally substituted heteroC 4-20 alkynyl. In some embodiments, R L is optionally substituted heteroC 4-19 alkynyl. In some embodiments, R L is optionally substituted heteroC 4-18 alkynyl. In some embodiments, R L is optionally substituted heteroC 4-17 alkynyl. In some embodiments, R L is optionally substituted heteroC 4-16 alkynyl. In some embodiments, R L is optionally substituted heteroC 4-15 alkynyl. In some embodiments, R L is optionally substituted heteroC 4-14 alkynyl. In some embodiments, R L is optionally substituted heteroC 4-13 alkynyl. In some embodiments, R L is optionally substituted heteroC 4-12 alkynyl. In some embodiments, R L is optionally substituted heteroC 4-11 alkynyl. In some embodiments, R L is optionally substituted heteroC 4-10 alkynyl. In some embodiments, R L is optionally substituted heteroC 4-9 alkynyl. In some embodiments, R L is optionally substituted heteroC 4-8 alkynyl. In some embodiments, R L is optionally substituted heteroC 4-7 alkynyl. In some embodiments, R L is optionally substituted heteroC 4-6 alkynyl.

In some embodiments, R L is optionally substituted heteroC 6-50 alkynyl. In some embodiments, R L is optionally substituted heteroC 6-40 alkynyl. In some embodiments, R L is optionally substituted heteroC 6-30 alkynyl. In some embodiments, R L is optionally substituted heteroC 6-20 alkynyl. In some embodiments, R L is optionally substituted heteroC 6-19 alkynyl. In some embodiments, R L is optionally substituted heteroC 6-18 alkynyl. In some embodiments, R L is optionally substituted heteroC 6-17 alkynyl. In some embodiments, R L is optionally substituted heteroC 6-16 alkynyl. In some embodiments, R L is optionally substituted heteroC 6-15 alkynyl. In some embodiments, R L is optionally substituted heteroC 6-14 alkynyl. In some embodiments, R L is optionally substituted heteroC 6-13 alkynyl. In some embodiments, R L is optionally substituted heteroC 6-12 alkynyl. In some embodiments, R L is optionally substituted heteroC 6-11 alkynyl. In some embodiments, R L is optionally substituted heteroC 6-10 alkynyl. In some embodiments, R L is optionally substituted heteroC 6-9 alkynyl. In some embodiments, R L is optionally substituted heteroC 6-8 alkynyl. In some embodiments, R L is optionally substituted heteroC 6-7 alkynyl.

In some embodiments, R L is optionally substituted heteroC 8-50 alkynyl. In some embodiments, R L is optionally substituted heteroC 8-40 alkynyl. In some embodiments, R L is optionally substituted heteroC 8-30 alkynyl. In some embodiments, R L is optionally substituted heteroC 8-20 alkynyl. In some embodiments, R L is optionally substituted heteroC 8-19 alkynyl. In some embodiments, R L is optionally substituted heteroC 8-18 alkynyl. In some embodiments, R L is optionally substituted heteroC 8-17 alkynyl. In some embodiments, R L is optionally substituted heteroC 8-16 alkynyl. In some embodiments, R L is optionally substituted heteroC 8-15 alkynyl. In some embodiments, R L is optionally substituted heteroC 8-14 alkynyl. In some embodiments, R L is optionally substituted heteroC 8-13 alkynyl. In some embodiments, R L is optionally substituted heteroC 8-12 alkynyl. In some embodiments, R L is optionally substituted heteroC 8-11 alkynyl. In some embodiments, R L is optionally substituted heteroC 8-10 alkynyl. In some embodiments, R L is optionally substituted heteroC 8-9 alkynyl.

In some embodiments, R L is optionally substituted heteroC 9-50 alkynyl. In some embodiments, R L is optionally substituted heteroC 9-40 alkynyl. In some embodiments, R L is optionally substituted heteroC 9-30 alkynyl. In some embodiments, R L is optionally substituted heteroC 9-20 alkynyl. In some embodiments, R L is optionally substituted heteroC 9-19 alkynyl. In some embodiments, R L is optionally substituted heteroC 9-18 alkynyl. In some embodiments, R L is optionally substituted heteroC 9-17 alkynyl. In some embodiments, R L is optionally substituted heteroC 9-16 alkynyl. In some embodiments, R L is optionally substituted heteroC 9-15 alkynyl. In some embodiments, R L is optionally substituted heteroC 9-14 alkynyl. In some embodiments, R L is optionally substituted heteroC 9-13 alkynyl. In some embodiments, R L is optionally substituted heteroC 9-12 alkynyl. In some embodiments, R L is optionally substituted heteroC 9-11 alkynyl. In some embodiments, R L is optionally substituted heteroC 9-10 alkynyl.

In some embodiments, R L is optionally substituted heteroC 10-50 alkynyl. In some embodiments, R L is optionally substituted heteroC 10-40 alkynyl. In some embodiments, R L is optionally substituted heteroC 10-30 alkynyl. In some embodiments, R L is optionally substituted heteroC 10-20 alkynyl. In some embodiments, R L is optionally substituted heteroC 10-19 alkynyl. In some embodiments, R L is optionally substituted heteroC 10-18 alkynyl. In some embodiments, R L is optionally substituted heteroC 10-17 alkynyl. In some embodiments, R L is optionally substituted heteroC 10-16 alkynyl. In some embodiments, R L is optionally substituted heteroC 10-15 alkynyl. In some embodiments, R L is optionally substituted heteroC 10-14 alkynyl. In some embodiments, R L is optionally substituted heteroC 10-13 alkynyl. In some embodiments, R L is optionally substituted heteroC 10-12 alkynyl. In some embodiments, R L is optionally substituted heteroC 10-11 alkynyl.

In some embodiments, R L is optionally substituted heteroC 10-50 alkynyl. In some embodiments, R L is optionally substituted heteroC 10-40 alkynyl. In some embodiments, R L is optionally substituted heteroC 10-30 alkynyl. In some embodiments, R L is optionally substituted heteroC 10-20 alkynyl. In some embodiments, R L is optionally substituted heteroC 11-19 alkynyl. In some embodiments, R L is optionally substituted heteroC 11-18 alkynyl. In some embodiments, R L is optionally substituted heteroC 11-17 alkynyl. In some embodiments, R L is optionally substituted heteroC 11-16 alkynyl. In some embodiments, R L is optionally substituted heteroC 11-15 alkynyl. In some embodiments, R L is optionally substituted heteroC 11-14 alkynyl. In some embodiments, R L is optionally substituted heteroC 11-13 alkynyl. In some embodiments, R L is optionally substituted heteroC 11-12 alkynyl.

In some embodiments, R L is optionally substituted heteroC 12-50 alkynyl. In some embodiments, R L is optionally substituted heteroC 12-40 alkynyl. In some embodiments, R L is optionally substituted heteroC 12-30 alkynyl. In some embodiments, R L is optionally substituted heteroC 12-20 alkynyl. In some embodiments, R L is optionally substituted heteroC 12-19 alkynyl. In some embodiments, R L is optionally substituted heteroC 12-18 alkynyl. In some embodiments, R L is optionally substituted heteroC 12-17 alkynyl. In some embodiments, R L is optionally substituted heteroC 12-16 alkynyl. In some embodiments, R L is optionally substituted heteroC 12-15 alkynyl. In some embodiments, R L is optionally substituted heteroC 12-14 alkynyl. In some embodiments, R L is optionally substituted heteroC 12-13 alkynyl.

In some embodiments, R L is optionally substituted heteroC 6 alkynyl. In some embodiments, R L is optionally substituted heteroC 7 alkynyl. In some embodiments, R L is optionally substituted heteroC 8 alkynyl. In some embodiments, R L is optionally substituted heteroC 9 alkynyl. In some embodiments, R L is optionally substituted heteroC 10 alkynyl. In some embodiments, R L is optionally substituted heteroC 11 alkynyl. In some embodiments, R L is optionally substituted heteroC 12 alkynyl. In some embodiments, R L is optionally substituted heteroC 13 alkynyl. In some embodiments, R L is optionally substituted heteroC 14 alkynyl. In some embodiments, R L is optionally substituted heteroC 15 alkynyl. In some embodiments, R L is optionally substituted heteroC 16 alkynyl. In some embodiments, R L is optionally substituted heteroC 17 alkynyl. In some embodiments, R L is optionally substituted heteroC 18 alkynyl. In some embodiments, R L is optionally substituted heteroC 19 alkynyl. In some embodiments, R L is optionally substituted heteroC 20 alkynyl.

In some embodiments, for example, in any of the above embodiments, R L is a substituted heteroalkynyl group. In some embodiments, R L is an unsubstituted heteroalkynyl group. In some embodiments, R L is an optionally substituted straight-chain heteroalkynyl group. In some embodiments, R L is a substituted straight-chain heteroalkynyl group. In some embodiments, R L is an unsubstituted straight-chain heteroalkynyl group. In some embodiments, R L is an optionally substituted branched heteroalkynyl group. In some embodiments, R L is a substituted branched heteroalkynyl group. In some embodiments, R L is an unsubstituted branched heteroalkynyl group.

In some embodiments, R L is a polymer. As used herein, a “polymer”, in some embodiments, refers to a compound comprised of at least 3 (e.g., at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, etc.) repeating covalently bound structural units. The polymer is in certain embodiments biocompatible (i.e., non-toxic). Exemplary polymers include, but are not limited to, cellulose polymers (e.g., hydroxyethylcellulose, ethylcellulose, carboxymethylcellulose, methylc cellulose, hydroxypropylmethylcellulose (HPMC)), dextran polymers, polymaleic acid polymers, poly(acrylic acid) polymers, poly(vinylalcohol) polymers, polyvinylpyrrolidone (PVP) polymers, and polyethyleneglycol (PEG) polymers, and combinations thereof.

In some embodiments, R L is a lipophilic, hydrophobic and/or non-polar group. In some embodiments, R L is a lipophilic group. In some embodiments, R L is a hydrophobic group. In some embodiments, R L is a non-polar group.

In some embodiments, when an R L group is depicted as bisecting a carbon-carbon bond, e.g., of the formula (i), it is understood that R L may be bonded to either carbon.

In some embodiments, at least one instance of R Q , R 2 , R 6 , or R 7 is a group of the formula (i), (ii), or (iii). In some embodiments, at least one instance of R 6 or R 7 of R 1 is a group of formula (i), (ii) or (iii). In some embodiments, at least one instance of R 6 Or R 7 of R 1 is a group of formula (i). In some embodiments, at least one instance of R 6 or R 7 of R 1 is a group of formula (i-a). In some embodiments, at least one instance of R 6 Or R 7 of R 1 is a group of formula (i-al). In some embodiments, at least one instance of R 6 Or R 7 of R 1 is a group of formula (i-b). In some embodiments, at least one instance of R 6 Or R 7 of R 1 is a group of formula (ii). In some embodiments, at least one instance of R 6 Or R 7 of R 1 is a group of formula (iii).

Various combinations of the above embodiments of Formula I are contemplated herein.

In some embodiments, wherein each instance of Q is O, the compound of formula I is a compound of formula I-a:

or a pharmaceutically acceptable salt thereof, wherein each variable is independently as defined above and described herein.

In certain embodiments, at least one R 1 is a group of formula (iv). In certain embodiments, each instance of R 1 is a group of formula (iv). In certain embodiments, each instance of R 2 is independently hydrogen or optionally substituted C 1-6 alkyl. In certain embodiments, each instance of R 2 is hydrogen. In certain embodiments, at least one instance of R 2 is a group of formula (i). In certain embodiments, at least one instance of R 2 is a group of formula (ii). In certain embodiments, at least one instance of R 2 is a group of formula (iii). In certain embodiments, p is 1. In certain embodiments, p is 2. In certain embodiments, p is 3.

In some embodiments, wherein at least one R 1 is a group of formula (iv), a compound of formula I is a compound of formula I-b:

or a pharmaceutically acceptable salt thereof, wherein each variable is independently as defined above and described herein.

In certain embodiments, each instance of R 1 is a group of formula (iv). In certain embodiments, each instance of R 2 is independently hydrogen or optionally substituted C 1-6 alkyl. In certain embodiments, each instance of R 2 is hydrogen. In certain embodiments, at least one instance of R 2 is a group of formula (i). In certain embodiments, at least one instance of R 2 is a group of formula (ii). In certain embodiments, at least one instance of R 2 is a group of formula (iii). In certain embodiments, p is 1. In certain embodiments, p is 2. In certain embodiments, p is 3. In certain embodiments, L is an optionally substituted alkylene. In certain embodiments, R 6 is a group of formula (i). In certain embodiments, R 6 is a group of formula (ii). In certain embodiments, R 6 is a group of formula (iii). In certain embodiments, R 7 is a group of formula (i). In certain embodiments, R 7 is a group of formula (ii). In certain embodiments, R 7 is a group of formula (iii). In certain embodiments, both R 6 and R 7 are independently groups of formula (i), (ii), or (iii).

In some embodiments, wherein each instance of R 1 is a group the formula (iv), a compound of Formula I is a compound of formula I-c:

or a pharmaceutically acceptable salt thereof, wherein each variable is independently as defined above and described herein.

In certain embodiments, each instance of R 2 is independently hydrogen or optionally substituted C 1-6 alkyl. In certain embodiments, each instance of R 2 is hydrogen. In certain embodiments, at least one instance of R 2 is a group of formula (i). In certain embodiments, at least one instance of R 2 is a group of formula (ii). In certain embodiments, at least one instance of R 2 is a group of formula (iii). In certain embodiments, p is 1. In certain embodiments, p is 2. In certain embodiments, p is 3. In certain embodiments, L is an optionally substituted alkylene. In certain embodiments, R 6 is a group of formula (i). In certain embodiments, R 6 is a group of formula (ii). In certain embodiments, R 6 is a group of formula (iii). In certain embodiments, R 7 is a group of formula (i). In certain embodiments, R 7 is a group of formula (ii). In certain embodiments, R 7 is a group of formula (iii). In certain embodiments, both R 6 and R 7 are independently groups of formula (i), (ii), or (iii).

In some embodiments, p=1. In some embodiments, a compound of formula I-c is a compound of formula I-c1:

or a pharmaceutically acceptable salt thereof, wherein each variable is independently as defined above and described herein.

In certain embodiments, each instance of R 2 is independently hydrogen or optionally substituted C 1-6 alkyl. In certain embodiments, each instance of R 2 is hydrogen. In certain embodiments, at least one instance of R 2 is a group of formula (i). In certain embodiments, at least one instance of R 2 is a group of formula (ii). In certain embodiments, at least one instance of R 2 is a group of formula (iii). In certain embodiments, L is an optionally substituted alkylene. In certain embodiments, R 6 is a group of formula (i). In certain embodiments, R 6 is a group of formula (ii). In certain embodiments, R 6 is a group of formula (iii). In certain embodiments, R 7 is a group of formula (i). In certain embodiments, R 7 is a group of formula (ii). In certain embodiments, R 7 is a group of formula (iii). In certain embodiments, both R 6 and R 7 are independently groups of formula (i), (ii), or (iii). In some embodiments, R 6 and R 7 are the same group of formula (i). In some embodiments, R 6 and R 7 are the same group of formula (i-a). In some embodiments, R 6 and R 7 are the same group of formula (i-al). In some embodiments, R 6 and R 7 are the same group of formula (i-b). In some embodiments, R 6 and R 7 are the same group of formula (ii). In some embodiments, R 6 and R 7 are the same group of formula (iii).

In some embodiments, each instance of R 2 is hydrogen. In some embodiments, a compound of formula I-c is a compound of formula I-c2:

or a pharmaceutically acceptable salt thereof, wherein each variable is independently as defined above and described herein.

In certain embodiments, L is an optionally substituted alkylene. In certain embodiments, R 6 is a group of formula (i). In certain embodiments, R 6 is a group of formula (ii). In certain embodiments, R 6 is a group of formula (iii). In certain embodiments, R 7 is a group of formula (i). In certain embodiments, R 7 is a group of formula (ii). In certain embodiments, R 7 is a group of formula (iii). In certain embodiments, both R 6 and R 7 are independently groups of formula (i), (ii), or (iii). In some embodiments, R 6 and R 7 are the same group of formula (i). In some embodiments, R 6 and R 7 are the same group of formula (i-a). In some embodiments, R 6 and R 7 are the same group of formula (i-al). In some embodiments, R 6 and R 7 are the same group of formula (i-b). In some embodiments, R 6 and R 7 are the same group of formula (ii). In some embodiments, R 6 and R 7 are the same group of formula (iii).

In some embodiments, L is an optionally substituted alkylene. In some embodiments, a compound of formula I-c is a compound of formula I-c3:

or a pharmaceutically acceptable salt thereof, wherein each variable is independently as defined above and described herein.

In some embodiments, q is an integer between 1 and 10, inclusive. In certain embodiments, R 6 is a group of formula (i). In certain embodiments, R 6 is a group of formula (ii). In certain embodiments, R 6 is a group of formula (iii). In certain embodiments, R 7 is a group of formula (i). In certain embodiments, R 7 is a group of formula (ii). In certain embodiments, R 7 is a group of formula (iii). In certain embodiments, both R 6 and R 7 are independently groups of formula (i), (ii), or (iii). In some embodiments, R 6 and R 7 are the same group of formula (i). In some embodiments, R 6 and R 7 are the same group of formula (i-a). In some embodiments, R 6 and R 7 are the same group of formula (i-al). In some embodiments, R 6 and R 7 are the same group of formula (i-b). In some embodiments, R 6 and R 7 are the same group of formula (ii). In some embodiments, R 6 and R 7 are the same group of formula (iii).

In some embodiments, a compound of formula I is a compound of formula I-d:

wherein each of p, R 2 and R L is independently as defined above and described herein.

In some embodiments, a compound of formula I is a compound of formula I-e:

wherein each of R 2 and R L is independently as defined above and described herein.

In some embodiments, a compound of formula I is a compound of formula I-f:

wherein each of R 2 and R L is independently as defined above and described herein.

In some embodiments, provided liposomes include a cationic lipid described in WO 2013063468 and in U.S. provisional application entitled “Lipid Formulations for Delivery of Messenger RNA” filed concurrently with the present application on even date, both of which are incorporated by reference herein. In some embodiments, a compound of formula I is a compound of formula I-c1-a:

or a pharmaceutically acceptable salt thereof, wherein:

each R 2 independently is hydrogen or C 1-3 alkyl;

each q independently is 2 to 6;

each R′ independently is hydrogen or C 1-3 alkyl;

and each R L independently is C 8-12 alkyl.

In some embodiments, each R 2 independently is hydrogen, methyl or ethyl. In some embodiments, each R 2 independently is hydrogen or methyl. In some embodiments, each R 2 is hydrogen.

In some embodiments, each q independently is 3 to 6. In some embodiments, each q independently is 3 to 5. In some embodiments, each q is 4.

In some embodiments, each R′ independently is hydrogen, methyl or ethyl. In some embodiments, each R′ independently is hydrogen or methyl. In some embodiments, each R′ independently is hydrogen.

In some embodiments, each R L independently is C 8-12 alkyl. In some embodiments, each R L independently is n-C 8-12 alkyl. In some embodiments, each R L independently is C 9-11 alkyl. In some embodiments, each R L independently is n-C 9-11 alkyl. In some embodiments, each R L independently is C 10 alkyl. In some embodiments, each R L independently is n-C 10 alkyl.

In some embodiments, each R 2 independently is hydrogen or methyl; each q independently is 3 to 5; each R′ independently is hydrogen or methyl; and each R L independently is C 8-12 alkyl.

In some embodiments, each R 2 is hydrogen; each q independently is 3 to 5; each R′ is hydrogen; and each R L independently is C 8-12 alkyl.

In some embodiments, each R 2 is hydrogen; each q is 4; each R′ is hydrogen; and each R L independently is C 8-12 alkyl.

In some embodiments, a compound of formula I is a compound of formula I-g:

wherein each of R L is independently as defined above and described herein.

In some embodiments, a compound of formula I is a compound of formula X:

or a pharmaceutically acceptable salt thereof, wherein each variable is independently as defined above and described herein.

In some embodiments, a compound of formula I is a compound of formula X-1:

or a pharmaceutically acceptable salt thereof, wherein each R 2 is independently as defined above and described herein.

In some embodiments, a compound of formula I is

or a pharmaceutically acceptable salt thereof.

Additional examples of cationic lipids suitable for the present invention are described in WO 2013063468, which is incorporated by reference herein in its entirety.

Chemical Definitions

Definitions of specific functional groups and chemical terms are described in more detail below. The chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th Ed., inside cover, and specific functional groups are generally defined as described therein. Additionally, general principles of organic chemistry, as well as specific functional moieties and reactivity, are described in Organic Chemistry , Thomas Sorrell, University Science Books, Sausalito, 1999; Smith and March March's Advanced Organic Chemistry, 5th Edition, John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive Organic Transformations , VCH Publishers, Inc., New York, 1989; and Carruthers, Some Modern Methods of Organic Synthesis, 3rd Edition, Cambridge University Press, Cambridge, 1987.

Compounds described herein can comprise one or more asymmetric centers, and thus can exist in various isomeric forms, e.g., enantiomers and/or diastereomers. For example, the compounds described herein can be in the form of an individual enantiomer, diastereomer or geometric isomer, or can be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomer. Isomers can be isolated from mixtures by methods known to those skilled in the art, including chiral high performance liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers can be prepared by asymmetric syntheses. See, for example, Jacques et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel, E. L. Stereochemistry of Carbon Compounds (McGraw-Hill, N Y, 1962); and Wilen, S. H. Tables of Resolving Agents and Optical Resolutions p. 268 (E. L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, Ind. 1972). The invention additionally contemplates compounds as individual isomers substantially free of other isomers, and alternatively, as mixtures of various isomers.

When a range of values is listed, it is intended to encompass each value and sub-range within the range. For example “C1-6 alkyl” is intended to encompass, C1, C2, C3, C4, C5, C6, C1-6, C1-5, C1-4, C1-3, C1-2, C2-6, C2-5, C2-4, C2-3, C3-6, C3-5, C3-4, C4-6, C4-5, and C5-6 alkyl.

As used herein, “alkyl” refers to a radical of a straight-chain or branched saturated hydrocarbon group having from 1 to 50 carbon atoms (“C1-50 alkyl”). In some embodiments, an alkyl group has 1 to 40 carbon atoms (“C1-40 alkyl”). In some embodiments, an alkyl group has 1 to 30 carbon atoms (“C1-30 alkyl”). In some embodiments, an alkyl group has 1 to 20 carbon atoms (“C1-20 alkyl”). In some embodiments, an alkyl group has 1 to 10 carbon atoms (“C1-10 alkyl”). In some embodiments, an alkyl group has 1 to 9 carbon atoms (“C1-9 alkyl”). In some embodiments, an alkyl group has 1 to 8 carbon atoms (“C1-8 alkyl”). In some embodiments, an alkyl group has 1 to 7 carbon atoms (“C1-7 alkyl”). In some embodiments, an alkyl group has 1 to 6 carbon atoms (“C1-6 alkyl”). In some embodiments, an alkyl group has 1 to 5 carbon atoms (“C1-5 alkyl”). In some embodiments, an alkyl group has 1 to 4 carbon atoms (“C1-4 alkyl”). In some embodiments, an alkyl group has 1 to 3 carbon atoms (“C1-3 alkyl”). In some embodiments, an alkyl group has 1 to 2 carbon atoms (“C1-2 alkyl”). In some embodiments, an alkyl group has 1 carbon atom (“C1 alkyl”). In some embodiments, an alkyl group has 2 to 6 carbon atoms (“C2-6 alkyl”). Examples of C1-6 alkyl groups include, without limitation, methyl (C1), ethyl (C2), n-propyl (C3), isopropyl (C3), n-butyl (C4), tert-butyl (C4), sec-butyl (C4), iso-butyl (C4), n-pentyl (C5), 3-pentanyl (C5), amyl (C5), neopentyl (C5), 3-methyl-2-butanyl (C5), tertiary amyl (C5), and n-hexyl (C6). Additional examples of alkyl groups include n-heptyl (C7), n-octyl (C8) and the like. Unless otherwise specified, each instance of an alkyl group is independently unsubstituted (an “unsubstituted alkyl”) or substituted (a “substituted alkyl”) with one or more substituents. In certain embodiments, the alkyl group is an unsubstituted C1-50 alkyl. In certain embodiments, the alkyl group is a substituted C1-50 alkyl.

As used herein, “heteroalkyl” refers to an alkyl group as defined herein which further includes at least one heteroatom (e.g., 1 to 25, e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus within (i.e., inserted between adjacent carbon atoms of) and/or placed at one or more terminal position(s) of the parent chain. In certain embodiments, a heteroalkyl group refers to a saturated group having from 1 to 50 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC1-50 alkyl”). In certain embodiments, a heteroalkyl group refers to a saturated group having from 1 to 40 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC1-40 alkyl”). In certain embodiments, a heteroalkyl group refers to a saturated group having from 1 to 30 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC1-30 alkyl”). In certain embodiments, a heteroalkyl group refers to a saturated group having from 1 to 20 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC1-20 alkyl”). In certain embodiments, a heteroalkyl group refers to a saturated group having from 1 to 10 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC1-10 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 9 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC1-9 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 8 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC1-8 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 7 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC1-7 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 6 carbon atoms and 1 or more heteroatoms within the parent chain (“heteroC1-6 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 5 carbon atoms and 1 or 2 heteroatoms within the parent chain (“heteroC1-5 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 4 carbon atoms and 1 or 2 heteroatoms within the parent chain (“heteroC1-4 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 3 carbon atoms and 1 heteroatom within the parent chain (“heteroC1-3 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 to 2 carbon atoms and 1 heteroatom within the parent chain (“heteroC1-2 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 1 carbon atom and 1 heteroatom (“heteroC1 alkyl”). In some embodiments, a heteroalkyl group is a saturated group having 2 to 6 carbon atoms and 1 or 2 heteroatoms within the parent chain (“heteroC2-6 alkyl”). Unless otherwise specified, each instance of a heteroalkyl group is independently unsubstituted (an “unsubstituted heteroalkyl”) or substituted (a “substituted heteroalkyl”) with one or more substituents. In certain embodiments, the heteroalkyl group is an unsubstituted heteroC1-50 alkyl. In certain embodiments, the heteroalkyl group is a substituted heteroC1-50 alkyl.

As used herein, “alkenyl” refers to a radical of a straight-chain or branched hydrocarbon group having from 2 to 50 carbon atoms and one or more carbon-carbon double bonds (e.g., 1, 2, 3, or 4 double bonds) (“C2-50 alkenyl”). In some embodiments, an alkenyl group has 2 to 40 carbon atoms (“C2-40 alkenyl”). In some embodiments, an alkenyl group has 2 to 30 carbon atoms (“C2-30 alkenyl”). In some embodiments, an alkenyl group has 2 to 20 carbon atoms (“C2-20 alkenyl”). In some embodiments, an alkenyl group has 2 to 10 carbon atoms (“C2-10 alkenyl”). In some embodiments, an alkenyl group has 2 to 9 carbon atoms (“C2-9 alkenyl”). In some embodiments, an alkenyl group has 2 to 8 carbon atoms (“C2-8 alkenyl”). In some embodiments, an alkenyl group has 2 to 7 carbon atoms (“C2-7 alkenyl”). In some embodiments, an alkenyl group has 2 to 6 carbon atoms (“C2-6 alkenyl”). In some embodiments, an alkenyl group has 2 to 5 carbon atoms (“C2-5 alkenyl”). In some embodiments, an alkenyl group has 2 to 4 carbon atoms (“C2-4 alkenyl”). In some embodiments, an alkenyl group has 2 to 3 carbon atoms (“C2-3 alkenyl”). In some embodiments, an alkenyl group has 2 carbon atoms (“C2 alkenyl”). The one or more carbon-carbon double bonds can be internal (such as in 2-butenyl) or terminal (such as in 1-butenyl). Examples of C2-4 alkenyl groups include, without limitation, ethenyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1-butenyl (C4), 2-butenyl (C4), butadienyl (C4), and the like. Examples of C2-6 alkenyl groups include the aforementioned C2-4 alkenyl groups as well as pentenyl (C5), pentadienyl (C5), hexenyl (C6), and the like. Additional examples of alkenyl include heptenyl (C7), octenyl (C8), octatrienyl (C8), and the like. Unless otherwise specified, each instance of an alkenyl group is independently unsubstituted (an “unsubstituted alkenyl”) or substituted (a “substituted alkenyl”) with one or more substituents. In certain embodiments, the alkenyl group is an unsubstituted C2-50 alkenyl. In certain embodiments, the alkenyl group is a substituted C2-50 alkenyl.

As used herein, “heteroalkenyl” refers to an alkenyl group as defined herein which further includes at least one heteroatom (e.g., 1 to 25, e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus within (i.e., inserted between adjacent carbon atoms of) and/or placed at one or more terminal position(s) of the parent chain. In certain embodiments, a heteroalkenyl group refers to a group having from 2 to 50 carbon atoms, at least one double bond, and 1 or more heteroatoms within the parent chain (“heteroC2-50 alkenyl”). In certain embodiments, a heteroalkenyl group refers to a group having from 2 to 40 carbon atoms, at least one double bond, and 1 or more heteroatoms within the parent chain (“heteroC2-40 alkenyl”). In certain embodiments, a heteroalkenyl group refers to a group having from 2 to 30 carbon atoms, at least one double bond, and 1 or more heteroatoms within the parent chain (“heteroC2-30 alkenyl”). In certain embodiments, a heteroalkenyl group refers to a group having from 2 to 20 carbon atoms, at least one double bond, and 1 or more heteroatoms within the parent chain (“heteroC2-20 alkenyl”). In certain embodiments, a heteroalkenyl group refers to a group having from 2 to 10 carbon atoms, at least one double bond, and 1 or more heteroatoms within the parent chain (“heteroC2-10 alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 9 carbon atoms at least one double bond, and 1 or more heteroatoms within the parent chain (“heteroC2-9 alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 8 carbon atoms, at least one double bond, and 1 or more heteroatoms within the parent chain (“heteroC2-8 alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 7 carbon atoms, at least one double bond, and 1 or more heteroatoms within the parent chain (“heteroC2-7 alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 6 carbon atoms, at least one double bond, and 1 or more heteroatoms within the parent chain (“heteroC2-6 alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 5 carbon atoms, at least one double bond, and 1 or 2 heteroatoms within the parent chain (“heteroC2-5 alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 4 carbon atoms. at least one double bond, and for 2 heteroatoms within the parent chain (“heteroC2-4 alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 3 carbon atoms, at least one double bond, and 1 heteroatom within the parent chain (“heteroC2-3 alkenyl”). In some embodiments, a heteroalkenyl group has 2 to 6 carbon atoms, at least one double, bond, and 1 or 2 heteroatoms within the parent chain (“heteroC2-6 alkenyl”). Unless otherwise specified, each instance of a heteroalkenyl group is independently unsubstituted (an “unsubstituted heteroalkenyl”) or substituted (a “substituted heteroalkenyl”) with one or more substituents. In certain embodiments, the heteroalkenyl group is an unsubstituted heteroC2-50 alkenyl. In certain embodiments, the heteroalkenyl group is a substituted heteroC2-50 alkenyl.

As used herein, “alkynyl” refers to a radical of a straight-chain or branched hydrocarbon group having from 2 to 50 carbon atoms and one or more carbon-carbon triple bonds (e.g., 1, 2, 3, or 4 triple bonds) and optionally one or more double bonds (e.g., 1, 2, 3, or 4 double bonds) (“C2-50 alkynyl”). An alkynyl group that has one or more triple bonds and one or more double bonds is also referred to as an “ene-yne”. In some embodiments, an alkynyl group has 2 to 40 carbon atoms (“C2-40 alkynyl”). In some embodiments, an alkynyl group has 2 to 30 carbon atoms (“C2-30 alkynyl”). In some embodiments, an alkynyl group has 2 to 20 carbon atoms (“C2-20 alkynyl”). In some embodiments, an alkynyl group has 2 to 10 carbon atoms (“C2-10 alkynyl”). In some embodiments, an alkynyl group has 2 to 9 carbon atoms (“C2-9 alkynyl”). In some embodiments, an alkynyl group has 2 to 8 carbon atoms (“C2-8 alkynyl”). In some embodiments, an alkynyl group has 2 to 7 carbon atoms (“C2-7 alkynyl”). In some embodiments, an alkynyl group has 2 to 6 carbon atoms (“C2-6 alkynyl”). In some embodiments, an alkynyl group has 2 to 5 carbon atoms (“C2-5 alkynyl”). In some embodiments, an alkynyl group has 2 to 4 carbon atoms (“C2-4 alkynyl”). In some embodiments, an alkynyl group has 2 to 3 carbon atoms (“C2-3 alkynyl”). In some embodiments, an alkynyl group has 2 carbon atoms (“C2 alkynyl”). The one or more carbon—carbon triple bonds can be internal (such as in 2-butynyl) or terminal (such as in 1-butynyl). Examples of C2-4 alkynyl groups include, without limitation, ethynyl (C2), 1-propynyl (C3), 2-propynyl (C3), 1-butynyl (C4), 2-butynyl (C4), and the like. Examples of C2-6 alkenyl groups include the aforementioned C2-4 alkynyl groups as well as pentynyl (C5), hexynyl (C6), and the like. Additional examples of alkynyl include heptynyl (C7), octynyl (C8), and the like. Unless otherwise specified, each instance of an alkynyl group is independently unsubstituted (an “unsubstituted alkynyl”) or substituted (a “substituted alkynyl”) with one or more substituents. In certain embodiments, the alkynyl group is an unsubstituted C2-50 alkynyl. In certain embodiments, the alkynyl group is a substituted C2-50 alkynyl.

As used herein, “heteroalkynyl” refers to an alkynyl group as defined herein which further includes at least one heteroatom (e.g., 1 to 25, e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus within (i.e., inserted between adjacent carbon atoms of) and/or placed at one or more terminal position(s) of the parent chain. In certain embodiments, a heteroalkynyl group refers to a group having from 2 to 50 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“heteroC2-50 alkynyl”). In certain embodiments, a heteroalkynyl group refers to a group having from 2 to 40 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“heteroC2-40 alkynyl”). In certain embodiments, a heteroalkynyl group refers to a group having from 2 to 30 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“heteroC2-30 alkynyl”). In certain embodiments, a heteroalkynyl group refers to a group having from 2 to 20 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“heteroC2-20 alkynyl”). In certain embodiments, a heteroalkynyl group refers to a group having from 2 to 10 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“heteroC2-10 alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 9 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“heteroC2-9 alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 8 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“heteroC2-8 alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 7 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“heteroC2-7 alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 6 carbon atoms, at least one triple bond, and 1 or more heteroatoms within the parent chain (“heteroC2-6 alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 5 carbon atoms, at least one triple bond, and 1 or 2 heteroatoms within the parent chain (“heteroC2-5 alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 4 carbon atoms, at least one triple bond, and for 2 heteroatoms within the parent chain (“heteroC2-4 alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 3 carbon atoms, at least one triple bond, and 1 heteroatom within the parent chain (“heteroC2-3 alkynyl”). In some embodiments, a heteroalkynyl group has 2 to 6 carbon atoms, at least one triple bond. and 1 or 2 heteroatoms within the parent chain (“heteroC2-6 alkynyl”). Unless otherwise specified, each instance of a heteroalkynyl group is independently unsubstituted (an “unsubstituted heteroalkynyl”) or substituted (a “substituted heteroalkynyl”) with one or more substituents. In certain embodiments, the heteroalkynyl group is an unsubstituted heteroC2-50 alkynyl. In certain embodiments, the heteroalkynyl group is a substituted heteroC2-50 alkynyl.

As used herein, “carbocyclyl” or “carbocyclic” refers to a radical of a non-aromatic cyclic hydrocarbon group having from 3 to 10 ring carbon atoms (“C3-10 carbocyclyl”) and zero heteroatoms in the non-aromatic ring system. In some embodiments, a carbocyclyl group has 3 to 8 ring carbon atoms (“C3-8 carbocyclyl”). In some embodiments, a carbocyclyl group has 3 to 7 ring carbon atoms (“C3-7 carbocyclyl”). In some embodiments, a carbocyclyl group has 3 to 6 ring carbon atoms (“C3-6 carbocyclyl”). In some embodiments, a carbocyclyl group has 4 to 6 ring carbon atoms (“C4-6 carbocyclyl”). In some embodiments, a carbocyclyl group has 5 to 6 ring carbon atoms (“C5-6 carbocyclyl”). In some embodiments, a carbocyclyl group has 5 to 10 ring carbon atoms (“C5-10 carbocyclyl”). Exemplary C3-6 carbocyclyl groups include, without limitation, cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (C6), cyclohexenyl (C6), cyclohexadienyl (C6), and the like. Exemplary C3-8 carbocyclyl groups include, without limitation, the aforementioned C3-6 carbocyclyl groups as well as cycloheptyl (C7), cycloheptenyl (C7), cycloheptadienyl (C7), cycloheptatrienyl (C7), cyclooctyl (C8), cyclooctenyl (C8), bicyclo[2.2.1]heptanyl (C7), bicyclo[2.2.2]octanyl (C8), and the like. Exemplary C3-10 carbocyclyl groups include, without limitation, the aforementioned C3-8 carbocyclyl groups as well as cyclononyl (C9), cyclononenyl (C9), cyclodecyl (C10), cyclodecenyl (C10), octahydro-1H-indenyl (C9), decahydronaphthalenyl (C10), spiro[4.5]decanyl (C10), and the like. As the foregoing examples illustrate, in certain embodiments, the carbocyclyl group is either monocyclic (“monocyclic carbocyclyl”) or polycyclic (e.g., containing a fused, bridged or spiro ring system such as a bicyclic system (“bicyclic carbocyclyl”) or tricyclic system (“tricyclic carbocyclyl”)) and can be saturated or can contain one or more carbon-carbon double or triple bonds. “Carbocyclyl” also includes ring systems wherein the carbocyclyl ring, as defined above, is fused with one or more aryl or heteroaryl groups wherein the point of attachment is on the carbocyclyl ring, and in such instances, the number of carbons continue to designate the number of carbons in the carbocyclic ring system. Unless otherwise specified, each instance of a carbocyclyl group is independently unsubstituted (an “unsubstituted carbocyclyl”) or substituted (a “substituted carbocyclyl”) with one or more substituents. In certain embodiments, the carbocyclyl group is an unsubstituted C3-10 carbocyclyl. In certain embodiments, the carbocyclyl group is a substituted C3-10 carbocyclyl.

In some embodiments, “carbocyclyl” or “carbocyclic” is referred to as a “cycloalkyl”, i.e., a monocyclic, saturated carbocyclyl group having from 3 to 10 ring carbon atoms (“C3-10 cycloalkyl”). In some embodiments, a cycloalkyl group has 3 to 8 ring carbon atoms (“C3-8 cycloalkyl”). In some embodiments, a cycloalkyl group has 3 to 6 ring carbon atoms (“C3-6, cycloalkyl”). In some embodiments, a cycloalkyl group has 4 to 6 ring carbon atoms (“C4-6 cycloalkyl”). In some embodiments, a cycloalkyl group has 5 to 6 ring carbon atoms (“C5-6 cycloalkyl”). In some embodiments, a cycloalkyl group has 5 to 10 ring carbon atoms (“C5-10 cycloalkyl”). Examples of C5-6 cycloalkyl groups include cyclopentyl (C5) and cyclohexyl (C5). Examples of C3-6 cycloalkyl groups include the aforementioned C5-6 cycloalkyl groups as well as cyclopropyl (C3) and cyclobutyl (C4). Examples of C3-8 cycloalkyl groups include the aforementioned C3-6 cycloalkyl groups as well as cycloheptyl (C7) and cyclooctyl (C8). Unless otherwise specified, each instance of a cycloalkyl group is independently unsubstituted (an “unsubstituted cycloalkyl”) or substituted (a “substituted cycloalkyl”) with one or more substituents. In certain embodiments, the cycloalkyl group is an unsubstituted C3-10 cycloalkyl. In certain embodiments, the cycloalkyl group is a substituted C3-10 cycloalkyl.

As used herein, “heterocyclyl” or “heterocyclic” refers to a radical of a 3- to 14-membered non-aromatic ring system having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4) ring heteroatoms, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“3-14 membered heterocyclyl”). In heterocyclyl groups that contain one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, as valency permits. A heterocyclyl group can either be monocyclic (“monocyclic heterocyclyl”) or polycyclic (e.g., a fused, bridged or spiro ring system such as a bicyclic system (“bicyclic heterocyclyl”) or tricyclic system (“tricyclic heterocyclyl”)). and can be saturated or can contain one or more carbon-carbon double or triple bonds. Heterocyclyl polycyclic ring systems can include one or more heteroatoms in one or both rings. “Heterocyclyl” also includes ring systems wherein the heterocyclyl ring, as defined above, is fused with one or more carbocyclyl groups wherein the point of attachment is either on the carbocyclyl or heterocyclyl ring, or ring systems wherein the heterocyclyl ring, as defined above, is fused with one or more aryl or heteroaryl groups, wherein the point of attachment is on the heterocyclyl ring, and in such instances, the number of ring members continue to designate the number of ring members in the heterocyclyl ring system. Unless otherwise specified, each instance of heterocyclyl is independently unsubstituted (an “unsubstituted heterocyclyl”) or substituted (a “substituted heterocyclyl”) with one or more substituents. In certain embodiments, the heterocyclyl group is an unsubstituted 3-14 membered heterocyclyl. In certain embodiments, the heterocyclyl group is a substituted 3-14 membered heterocyclyl.

In some embodiments, a heterocyclyl group is a 5-10 membered non-aromatic ring system having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4) ring heteroatoms, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5-10 membered heterocyclyl”). In some embodiments, a heterocyclyl group is a 5-8 membered non-aromatic ring system having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4) ring heteroatoms, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5-8 membered heterocyclyl”). In some embodiments, a heterocyclyl group is a 5-6 membered non-aromatic ring system having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4) ring heteroatoms, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5-6 membered heterocyclyl”). In some embodiments, the 5-6 membered heterocyclyl has 1 or more (e.g., 1, 2, or 3) ring heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. In some embodiments, the 5-6 membered heterocyclyl has 1 or 2 ring heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. In some embodiments, the 5-6 membered heterocyclyl has 1 ring heteroatom selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus.

Exemplary 3-membered heterocyclyl groups containing 1 heteroatom include, without limitation, azirdinyl, oxiranyl, thiorenyl. Exemplary 4-membered heterocyclyl groups containing 1 heteroatom include, without limitation, azetidinyl, oxetanyl and thietanyl. Exemplary 5-membered heterocyclyl groups containing 1 heteroatom include, without limitation. tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyl and pyrrolyl-2,5-dione. Exemplary 5-membered heterocyclyl groups containing 2 heteroatoms include, without limitation, dioxolanyl, oxathiolanyl and dithiolanyl. Exemplary 5-membered heterocyclyl groups containing 3 heteroatoms include, without limitation, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclyl groups containing 1 heteroatom include, without limitation, piperidinyl, tetrahydropyranyl, dihydropyridinyl, and thianyl. Exemplary 6-membered heterocyclyl groups containing 2 heteroatoms include, without limitation, piperazinyl, morpholinyl, dithianyl, dioxanyl. Exemplary 6-membered heterocyclyl groups containing 2 heteroatoms include, without limitation, triazinanyl. Exemplary 7-membered heterocyclyl groups containing 1 heteroatom include, without limitation, azepanyl, oxepanyl and thiepanyl. Exemplary 8-membered heterocyclyl groups containing 1 heteroatom include, without limitation, azocanyl, oxecanyl and thiocanyl. Exemplary bicyclic heterocyclyl groups include, without limitation, indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, tetrahydrobenzothienyl, tetrahydrobenzofuranyl, tetrahydroindolyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, decahydroisoquinolinyl, octahydrochromenyl, octahydroisochromenyl, decahydronaphthyridinyl, decahydro-1,8-naphthyridinyl, octahydropyrrolo[3,2-b]pyrrole, indolinyl, phthalimidyl, naphthalimidyl, chromanyl, chromenyl, 1H-benzo[e][1,4]diazepinyl, 1,4,5,7-tetrahydropyrano[3,4-b]pyrrolyl, 5,6-dihydro-4H-furo[3,2-b]pyrrolyl, 6,7-dihydro-5H-furo[3,2-b]pyranyl, 5,7-dihydro-4H-thieno[2,3-c]pyranyl, 2,3-dihydro-1H-pyrrolo[2,3-b]pyridinyl, 2,3-dihydrofuro[2,3-b]pyridinyl, 4,5,6,7-tetrahydro-1H-pyrrolo-[2,3-b]pyridinyl, 4,5,6,7-tetrahydrofuro[3,2-c]pyridinyl, 4,5,6,7-tetrahydrothieno [3,2-b]pyridinyl, 1,2,3,4-tetrahydro-1,6-naphthyridinyl, and the like.

As used herein, “aryl” refers to a radical of a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 π electrons shared in a cyclic array) having 6-14 ring carbon atoms and zero heteroatoms provided in the aromatic ring system (“C6-14 aryl”). In some embodiments, an aryl group has 6 ring carbon atoms (“C6 aryl”; e.g., phenyl). In some embodiments, an aryl group has 10 ring carbon atoms (“C10 aryl”; e.g., naphthyl such as 1-naphthyl and 2-naphthyl). In some embodiments, an aryl group has 14 ring carbon atoms (“C14 aryl”; e.g., anthracyl). “Aryl” also includes ring systems wherein the aryl ring, as defined above, is fused with one or more carbocyclyl or heterocyclyl groups wherein the radical or point of attachment is on the aryl ring, and in such instances, the number of carbon atoms continue to designate the number of carbon atoms in the aryl ring system. Unless otherwise specified, each instance of an aryl group is independently unsubstituted (an “unsubstituted aryl”) or substituted (a “substituted aryl”) with one or more substituents. In certain embodiments, the aryl group is an unsubstituted C6-14 aryl. In certain embodiments, the aryl group is a substituted C6-14 aryl.

As used herein, “heteroaryl” refers to a radical of a 5-14 membered monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 π electrons shared in a cyclic array) having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4 ring heteroatoms) ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5-14 membered heteroaryl”). In heteroaryl groups that contain one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, as valency permits. Heteroaryl polycyclic ring systems can include one or more heteroatoms in one or both rings. “Heteroaryl” includes ring systems wherein the heteroaryl ring, as defined above, is fused with one or more carbocyclyl or heterocyclyl groups wherein the point of attachment is on the heteroaryl ring, and in such instances, the number of ring members continue to designate the number of ring members in the heteroaryl ring system. “Heteroaryl” also includes ring systems wherein the heteroaryl ring, as defined above, is fused with one or more aryl groups wherein the point of attachment is either on the aryl or heteroaryl ring, and in such instances, the number of ring members designates the number of ring members in the fused polycyclic (aryl/heteroaryl) ring system. Polycyclic heteroaryl groups wherein one ring does not contain a heteroatom (e.g., indolyl, quinolinyl, carbazolyl, and the like) the point of attachment can be on either ring, i.e., either the ring bearing a heteroatom (e.g., 2-indolyl) or the ring that does not contain a heteroatom (e.g., 5-indolyl).

In some embodiments, a heteroaryl group is a 5-10 membered aromatic ring system having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4) ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5-10 membered heteroaryl”). In some embodiments, a heteroaryl group is a 5-8 membered aromatic ring system having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4) ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5-8 membered heteroaryl”). In some embodiments, a heteroaryl group is a 5-6 membered aromatic ring system having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4) ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5-6 membered heteroaryl”). In some embodiments, the 5-6 membered heteroaryl has 1 or more (e.g., 1, 2, or 3) ring heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. In some embodiments, the 5-6 membered heteroaryl has 1 or 2 ring heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. In some embodiments, the 5-6 membered heteroaryl has 1 ring heteroatom selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. Unless otherwise specified, each instance of a heteroaryl group is independently unsubstituted (an “unsubstituted heteroaryl”) or substituted (a “substituted heteroaryl”) with one or more substituents. In certain embodiments, the heteroaryl group is an unsubstituted 5-14 membered heteroaryl. In certain embodiments, the heteroaryl group is a substituted 5-14 membered heteroaryl.

Exemplary 5-membered heteroaryl groups containing 1 heteroatom include, without limitation, pyrrolyl, furanyl and thiophenyl. Exemplary 5-membered heteroaryl groups containing 2 heteroatoms include, without limitation, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroaryl groups containing 3 heteroatoms include, without limitation, triazolyl, oxadiazolyl, and thiadiazolyl. Exemplary 5-membered heteroaryl groups containing 4 heteroatoms include, without limitation, tetrazolyl. Exemplary 6-membered heteroaryl groups containing 1 heteroatom include, without limitation. pyridinyl. Exemplary 6-membered heteroaryl groups containing 2 heteroatoms include, without limitation, pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered heteroaryl groups containing 3 or 4 heteroatoms include, without limitation, triazinyl and tetrazinyl, respectively. Exemplary 7-membered heteroaryl groups containing 1 heteroatom include, without limitation, azepinyl, oxepinyl, and thiepinyl. Exemplary 5,6-bicyclic heteroaryl groups include, without limitation, indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzoisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzthiazolyl, benzisothiazolyl, benzthiadiazolyl, indolizinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryl groups include, without limitation, naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl. Exemplary tricyclic heteroaryl groups include, without limitation, phenanthridinyl, dibenzofuranyl, carbazolyl, acridinyl, phenothiazinyl, phenoxazinyl and phenazinyl.

As used herein, the term “partially unsaturated” refers to a ring moiety that includes at least one double or triple bond. The term “partially unsaturated” is intended to encompass rings having multiple sites of unsaturation, but is not intended to include aromatic groups (e.g., aryl or heteroaryl moieties) as herein defined.

As used herein, the term “saturated” refers to a ring moiety that does not contain a double or triple bond, i.e., the ring contains all single bonds.

Affixing the suffix “-ene” to a group indicates the group is a divalent moiety, e.g., alkylene is the divalent moiety of alkyl, alkenylene is the divalent moiety of alkenyl,

alkynylene is the divalent moiety of alkynyl, heteroalkylene is the divalent moiety of heteroalkyl, heteroalkenylene is the divalent moiety of heteroalkenyl, heteroalkynylene is the divalent moiety of heteroalkynyl, carbocyclylene is the divalent moiety of carbocyclyl, heterocyclylene is the divalent moiety of heterocyclyl, arylene is the divalent moiety of aryl, and heteroarylene is the divalent moiety of heteroaryl.

As understood from the above, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl groups, as defined herein, are, in certain embodiments, optionally substituted. Optionally substituted refers to a group which may be substituted or unsubstituted (e.g., “substituted” or “unsubstituted” alkyl, “substituted” or “unsubstituted” alkenyl, “substituted” or “unsubstituted” alkynyl, “substituted” or “unsubstituted” heteroalkyl, “substituted” or “unsubstituted” heteroalkenyl, “substituted” or “unsubstituted” heteroalkynyl. “substituted” or “unsubstituted” carbocyclyl. “substituted” or “unsubstituted” heterocyclyl, “substituted” or “unsubstituted” aryl or “substituted” or “unsubstituted” heteroaryl group). In general, the term “substituted” means that at least one hydrogen present on a group is replaced with a permissible substituent, e.g., a substituent which upon substitution results in a stable compound, e.g., a compound which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, or other reaction. Unless otherwise indicated, a “substituted” group has a substituent at one or more substitutable positions of the group, and when more than one position in any given structure is substituted, the substituent is either the same or different at each position. The term “substituted” is contemplated to include substitution with all permissible substituents of organic compounds, any of the substituents described herein that results in the formation of a stable compound. The present invention contemplates any and all such combinations in order to arrive at a stable compound. For purposes of this invention, heteroatoms such as nitrogen may have hydrogen substituents and/or any suitable substituent as described herein which satisfy the valencies of the heteroatoms and results in the formation of a stable moiety.

Exemplary carbon atom substituents include, but are not limited to, halogen, —CN, —NO2, —N3, —SO2H, —SO3H, —OH, —ORaa, —ON(Rbb)2, —N(Rbb)2, —N(Rbb)3+X—, —N(ORcc)Rbb, —SeH, —SeRaa, —SH, —SRaa, —SSRcc, —C(═O)Raa, —CO2H, —CHO, —C(ORcc)2, —CO2Raa, —OC(═O)Raa, —OCO2Raa, —C(═O)N(Rbb)2, —OC(═O)N(Rbb)2, —NRbbC(═O)Raa, —NRbbCO2Raa, —NRbbC(═O)N(Rbb)2, —C(═NRbb)Raa, —C(═NRbb)ORaa, —OC(═NRbb)Raa, —OC(═NRbb)ORaa, —C(═NRbb)N(Rbb)2, —OC(═NRbb)N(Rbb)2, —NRbbC(═NRbb)N(Rbb)2, —C(═O)NRbbSO2Raa, —NRbbSO2Raa, —SO2N(Rbb)2, —SO2Raa, —SO2ORaa, —OSO2Raa, —S(═O)Raa, —OS(═O)Raa, —Si(Raa)3-OSi(Raa)3-C(═S)N(Rbb)2, —C(═O)SRaa, —C(═S)SRaa, —SC(═S)SRaa, —SC(═O)SRaa, —OC(═O)SRaa, —SC(═O)ORaa, —SC(═O)Raa, —P(═O)2Raa, —OP(═O)2Raa, —P(═O)(Raa)2, —OP(═O)(Raa)2, —OP(═O)(ORcc)2, —P(═O)2N(Rbb)2, —OP(═O)2N(Rbb)2, —P(═O)(NRbb)2, —OP(═O)(NRbb)2, —NRbbP(═O)(ORcc)2, —NRbbP(═O)(NRbb)2, —P(Rcc)2, —P(Rcc)3, —OP(Rcc)2, —OP(Rcc)3, —B(Raa)2, —B(ORcc)2, —BRaa(ORcc), C1-50 alkyl, C2-50 alkenyl, C2-50 alkynyl, C3-14 carbocyclyl, 3-14 membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups;

or two geminal hydrogens on a carbon atom are replaced with the group ═O, ═S, ═NN(Rbb)2, ═NNRbbC(═O)Raa, ═NNRbbC(═O)ORaa, ═NNRbbS(═O)2Raa, ═NRbb, or ═NORcc;

each instance of Raa is, independently, selected from C1-50 alkyl, C2-50 alkenyl, C2-50 alkynyl, C3-10 carbocyclyl, 3-14 membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl, or two Raa groups are joined to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups;

each instance of Rbb is, independently, selected from hydrogen, —OH, —ORaa, —N(Rcc)2, —CN, —C(═O)Raa, —C(═O)N(Rcc)2, —CO2Raa, —SO2Raa, —C(═NRcc)ORaa, —C(═NRcc)N(Rcc)2, —SO2N(Rcc)2, —SO2Rcc, —SO20Rcc, —SORaa, —C(═S)N(Rcc)2, —C(═O)SRcc, —C(═S)SRcc, —P(═O)2Raa, —P(═O)(Raa)2, —P(═O)2N(Rcc)2, —P(═O)(NRcc)2, C1-50 alkyl, C2-50 alkenyl, C2-50 alkynyl, C3-10 carbocyclyl, 3-14 membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl, or two Rbb groups, together with the heteroatom to which they are attached, form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups;

each instance of Rcc is, independently, selected from hydrogen, C1-50 alkyl, C2-50 alkenyl, C2-50 alkynyl, C3-10 carbocyclyl, 3-14 membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl, or two Rcc groups, together with the heteroatom to which they are attached, form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups;

each instance of Rdd is, independently, selected from halogen, —CN, —NO2, —N3, —SO2H, —SO3H, —OH, —ORee, —ON(Rff)2, —N(Rff)2, —N(Rff)3+X—, —N(ORee)Rff, —SH, —SRee, —SSRee, —C(═O)Ree, —CO2H, —CO2Ree, —OC(═O)Ree, —OCO2Ree, —C(═O)N(Rff)2, —OC(═O)N(Rff)2, —NRffC(═O)Ree, —NRffCO2Ree, —NRffC(═O)N(Rff)2, —C(═NRff)ORee, —OC(═NRff)Ree, —OC(═NRff)ORee, —C(═NRff)N(Rff)2, —OC(═NRff)N(Rff)2, —NRffC(═NRff)N(Rff)2, —NRffSO2Ree, —S 02N(Rff)2, —SO2Ree, —SO2ORee, —OSO2Ree, —S(═O)Ree, —Si(Ree)3, —OSi(Ree)3, —C(═S)N(Rff)2, —C(═O)SRee, —C(═S)SRee, —SC(═S)SRee, —P(═O)2Ree, —P(═O)(Ree)2, —OP(═O)(Ree)2, —OP(═O)(ORee)2, C1-50 alkyl, C2-50 alkenyl, C2-50 alkynyl, C3-10 carbocyclyl, 3-10 membered heterocyclyl, C6-10 aryl, 5-10 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rgg groups, or two geminal Rdd substituents can be joined to form ═O or ═S;

each instance of Ree is, independently, selected from C1-50 alkyl, C2-50 alkenyl, C2-50 alkynyl, C3-10 carbocyclyl, C6-10 aryl, 3-10 membered heterocyclyl, and 3-10 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rgg groups;

each instance of Rff is, independently, selected from hydrogen, C1-50 alkyl, C2-50 alkenyl, C2-50 alkynyl, C3-10 carbocyclyl, 3-10 membered heterocyclyl, C6-10 aryl and 5-10 membered heteroaryl, or two Rff groups, together with the heteroatom to which they are attached, form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rgg groups; and

each instance of Rgg is, independently, halogen, —CN, —NO2, —N3, —SO2H, —SO3H, —OH, —OC1-50 alkyl, —ON(C1-50 alkyl)2, —N(C1-50 alkyl)2, —N(C1-50 alkyl)3+X—, —NH(C1-50 alkyl)2+X—, —NH2(C1-50 alkyl)+X—, —NH3+X—, —N(OC1-50 alkyl)(C1-50 alkyl), —N(OH)(C1-50 alkyl), —NH(OH), —SH, —SC1-50 alkyl, —SS(C1-50 alkyl), —C(═O)(C1-50 alkyl), —CO2H, —CO2(C1-50 alkyl), —OC(═O)(C1-50 alkyl), —OCO2(C1-50 alkyl), —C(═O)NH2, —C(═O)N(C1-50 alkyl)2, —OC(═O)NH(C1-50 alkyl), —NHC(═O)(C1-50 alkyl), —N(C1-50 alkyl)C(═O)(C1-50 alkyl), —NHCO2(C1-50 alkyl), —NHC(═O)N(C1-50 alkyl)2, —NHC(═O)NH(C1-50 alkyl), —NHC(═O)NH2, —C(═NH)O(C1-50 alkyl), —OC(═NH)(C1-50 alkyl), —OC(═NH)OC1-50 alkyl, —C(═NH)N(C1-50 alkyl)2, —C(═NH)NH(C1-50 alkyl), —C(═NH)NH2, —OC(═NH)N(C1-50alky 1)2, —OC(NH)NH(C1-50 alkyl), —OC(NH)NH2, —NHC(NH)N(C1-50 alkyl)2, —NHC(═NH)NH2, —NHSO2 (C1-50 alkyl), —SO2N(C1-50 alkyl)2, —SO2NH(C1-50 alkyl), —SO2NH2, —SO2C1-50 alkyl, —SO2OC1-50 alkyl, —OSO2C1-6 alkyl, —SOC1-6 alkyl, —Si(C1-50 alkyl)3, —OSi(C1-6 alkyl)3-C(═S)N(C1-50 alkyl)2, C(═S)NH(C1-50 alkyl), C(═S)NH2, —C(═O)S(C1-6 alkyl), —C(═S)SC1-6 alkyl, —SC(═S)SC1-6 alkyl, —P(═O)2(C1-50 alkyl), —P(═O)(C1-50 alkyl)2, —OP(═O)(C1-50 alkyl)2, —OP(═O)(OC1-50 alkyl)2, C1-50 alkyl, C2-50 alkenyl, C2-50 alkynyl, C3-10 carbocyclyl, C6-10 aryl, 3-10 membered heterocyclyl, 5-10 membered heteroaryl; or two geminal Rgg substituents can be joined to form ═O or ═S;

wherein X— is a counterion.

As used herein, the term “halo” or “halogen” refers to fluorine (fluoro, —F), chlorine (chloro, —Cl), bromine (bromo, —Br), or iodine (iodo, —I).

As used herein, a “counterion” is a negatively charged group associated with a positively charged quarternary amine in order to maintain electronic neutrality. Exemplary counterions include halide ions (e.g., F—, Cl—, Br—, I—), NO3-, ClO4-, OH—, H2PO4-, HSO4-, sulfonate ions (e.g., methansulfonate, trifluoromethanesulfonate, p-toluenesulfonate, benzenesulfonate, 10-camphor sulfonate, naphthalene-2-sulfonate, naphthalene-1-sulfonic acid-5-sulfonate, ethan-1-sulfonic acid-2-sulfonate, and the like), and carboxylate ions (e.g., acetate, ethanoate, propanoate, benzoate, glycerate, lactate, tartrate, glycolate, and the like).

Nitrogen atoms can be substituted or unsubstituted as valency permits, and include primary, secondary, tertiary, and quarternary nitrogen atoms. Exemplary nitrogen atom substitutents include, but are not limited to, hydrogen, —OH, —ORaa, —N(Rcc)2, —CN, —C(═O)Raa, —C(═O)N(Rcc)2, —CO2Raa, —SO2Raa, —C(═NRbb)Raa, —C(═NRcc)ORaa, —C(═NRcc)N(Rcc)2, —SO2N(Rcc)2, —SO2Rcc, —SO2ORcc, —SORaa, —C(═S)N(Rcc)2, —C(═O)SRcc, —C(═S)SRcc, —P(═O)2Raa, —P(═O)(Raa)2, —P(═O)2N(Rcc)2, —P(═O)(NRcc)2, C1-50 alkyl, C2-50 alkenyl, C2-50 alkynyl, C3-10 carbocyclyl, 3-14 membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl, or two Rcc groups, together with the N atom to which they are attached, form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups, and wherein Raa, Rbb, Rcc and Rdd are as defined above.

Nitrogen atoms can be substituted or unsubstituted as valency permits, and include primary, secondary, tertiary, and quarternary nitrogen atoms. Exemplary nitrogen atom substitutents include, but are not limited to, hydrogen, —OH, —ORaa, —N(Rcc)2, —CN, —C(═O)Raa, —C(═O)N(Rcc)2, —CO2Raa, —SO2Raa, —C(═NRbb)Raa, —C(═NRcc)ORaa, —C(═NRcc)N(Rcc)2, —SO2N(Rcc)2, —SO2Rcc, —SO2ORcc, —SORaa, —C(═S)N(Rcc)2, —C(═O)SRcc, —C(═S)SRcc, —P(═O)2Raa, —P(═O)(Raa)2, —P(═O)2N(Rcc)2, —P(═O)(NRcc)2, C1-10 alkyl, C1-10 perhaloalkyl, C2-10 alkenyl, C2-10 alkynyl, C3-10 carbocyclyl, 3-14 membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl, or two Rcc groups, together with the nitrogen atom to which they are attached, form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups, and wherein Raa, Rbb, Rcc and Rdd are as defined above.

In certain embodiments, the substituent present on a nitrogen atom is a nitrogen protecting group (also referred to as an amino protecting group). Nitrogen protecting groups include, but are not limited to, —OH, —ORaa, —N(Rcc)2, —C(═O)Raa, —C(═O)N(Rcc)2, —CO2Raa, —SO2Raa, —C(═NRcc)Raa, —C(═NRcc)ORaa, —C(═NRcc)N(Rcc)2, —SO2N(Rcc)2, —SO2Rcc, —SO2ORcc, —SORaa, —C(═S)N(Rcc)2, —C(═O)SRcc, —C(═S)SRcc, C1-10 alkyl (e.g., aralkyl, heteroaralkyl), C2-10 alkenyl, C2-10 alkynyl, C3-10 carbocyclyl, 3-14 membered heterocyclyl, C6-14 aryl, and 5-14 membered heteroaryl groups, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aralkyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4 or 5 Rdd groups, and wherein Raa, Rbb, Rcc and Rdd are as defined herein. Nitrogen protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis , T. W. Greene and P. G. M. Wuts, 3rd edition, John Wiley & Sons, 1999, incorporated herein by reference.

For example, nitrogen protecting groups such as amide groups (e.g., —C(═O)Raa) include, but are not limited to, formamide, acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide, phenylacetamide, 3-phenylpropanamide, picolinamide, 3-pyridylcarboxamide, N-benzoylphenylalanyl derivative, benzamide, p-phenylbenzamide, o-nitophenylacetamide, o-nitrophenoxyacetamide, acetoacetamide, (N′-dithiobenzyloxyacylamino)acetamide, 3-(p-hydroxyphenyl)propanamide, 3-(o-nitrophenyl)propanamide, 2-methyl-2-(o-nitrophenoxy)propanamide, 2-methyl-2-(o-phenylazophenoxy)propanamide, 4-chlorobutanamide, 3-methyl-3-nitrobutanamide, o-nitrocinnamide, N-acetylmethionine derivative, o-nitrobenzamide and o-(benzoyloxymethyl)benzamide.

Nitrogen protecting groups such as carbamate groups (e.g., —C(═O)ORaa) include, but are not limited to, methyl carbamate, ethyl carbamante, 9-fluorenylmethyl carbamate (Fmoc), 9-(2-sulfo)fluorenylmethyl carbamate, 9-(2,7-dibromo)fluoroenylmethyl carbamate, 2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methyl carbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc), 2,2,2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl carbamate (Teoc), 2-phenylethyl carbamate (hZ), 1-(1-adamantyl)-1-methylethyl carbamate (Adpoc), 1,1-dimethyl-2-haloethyl carbamate, 1,1-dimethyl-2,2-dibromoethyl carbamate (DB-t-BOC), 1,1-dimethyl-2,2,2-trichloroethyl carbamate (TCBOC), 1-methyl-1-(4-biphenylyl)ethyl carbamate (Bpoc), 1-(3,5-di-t-butylphenyl)-1-methylethyl carbamate (t-Bumeoc), 2-(2′- and 4′-pyridyl)ethyl carbamate (Pyoc), 2-(N,N-dicyclohexylcarboxamido)ethyl carbamate, t-butyl carbamate (BOC), 1-adamantyl carbamate (Adoc), vinyl carbamate (Voc), allyl carbamate (Alloc), 1-isopropylallyl carbamate (Ipaoc), cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate (Noc), 8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, alkyldithio carbamate, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz), p-nitobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzyl carbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzyl carbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl carbamate, 2-methylthioethyl carbamate, 2-methyl sulfonylethyl carbamate, 2-(p-toluenesulfonyl)ethyl carbamate, [2-(1,3-dithianyl)]methyl carbamate (Dmoc), 4-methylthiophenyl carbamate (Mtpc), 2,4-dimethylthiophenyl carbamate (Bmpc), 2-phosphonioethyl carbamate (Peoc), 2-triphenylphosphonioisopropyl carbamate (Ppoc), 1,1-dimethyl-2-cyanoethyl carbamate, m-chloro-p-acyloxybenzyl carbamate, p-(dihydroxyboryl)benzyl carbamate, 5-benzisoxazolylmethyl carbamate, 2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc), m-nitrophenyl carbamate, 3,5-dimethoxybenzyl carbamate, o-nitrobenzyl carbamate, 3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl(o-nitrophenyl)methyl carbamate, t-amyl carbamate, S-benzyl thiocarbamate, p-cyanobenzyl carbamate, cyclobutyl carbamate, cyclohexyl carbamate, cyclopentyl carbamate, cyclopropylmethyl carbamate, p-decyloxybenzyl carbamate, 2,2-dimethoxyacylvinyl carbamate, o-(N,N-dimethylcarboxamido)benzyl carbamate, 1,1-dimethyl-3-(N,N-dimethylcarboxamido)propyl carbamate, 1,1-dimethylpropynyl carbamate, di(2-pyridyl)methyl carbamate, 2-furanylmethyl carbamate, 2-iodoethyl carbamate, isoborynl carbamate, isobutyl carbamate, isonicotinyl carbamate, p-(p′-methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl carbamate, 1-methylcyclohexyl carbamate, 1-methyl-1-cyclopropylmethyl carbamate, 1-methyl-1(3,5-dimethoxyphenyl)ethyl carbamate, 1-methyl-1-(p-phenylazophenyl)ethyl carbamate, 1-methyl-1-phenylethyl carbamate, 1-methyl-1-(4-pyridyl)ethyl carbamate, phenyl carbamate, p-(phenylazo)benzyl carbamate, 2,4,6-tri-t-butylphenyl carbamate, 4-(trimethylammonium)benzyl carbamate, and 2,4,6-trimethylbenzyl carbamate.

Nitrogen protecting groups such as sulfonamide groups (e.g., —S(═O)2Raa) include, but are not limited to, p-toluenesulfonamide (Ts), benzenesulfonamide, 2,3,6,-trimethyl-4-methoxybenzenesulfonamide (Mtr), 2,4,6-trimethoxybenzenesulfonamide (Mtb), 2,6-dimethyl-4-methoxybenzenesulfonamide (Pme), 2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte), 4-methoxybenzenesulfonamide (Mbs), 2,4,6-trimethylbenzenesulfonamide (Mts), 2,6-dimethoxy-4-methylbenzenesulfonamide (iMds), 2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide (Ms), β-trimethyl silylethanesulfonamide (SES), 9-anthracenesulfonamide, 4-(4′,8′-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS), benzylsulfonamide, trifluoromethylsulfonamide, and phenacylsulfonamide.

Other nitrogen protecting groups include, but are not limited to, phenothiazinyl-(10)-acyl derivative, N′-p-toluenesulfonylaminoacyl derivative, N′-phenylaminothioacyl derivative, N-benzoylphenylalanyl derivative, N-acetylmethionine derivative, 4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts), N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole, N-1,1,4,4-tetramethyldisilylazacyclopentane adduct (STABASE), 5-substituted 1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted 1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, 1-substituted 3,5-dinitro-4-pyridone, N-methylamine, N-allylamine, N-[2-(trimethyl silyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine, N-(1-isopropyl-4-nitro-2-oxo-3-pyroolin-3-yl)amine, quaternary ammonium salts, N-benzylamine, N-di(4-methoxyphenyl)methylamine, N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr), N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr), N-9-phenylfluorenylamine (PhF), N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino (Fcm), N-2-picolylamino N′-oxide, N-1,1-dimethylthiomethyleneamine, N-benzylideneamine, N-p-methoxybenzylideneamine, N-diphenylmethyleneamine, N-[(2-pyridyl)mesityl]methyleneamine, N—(N′,N′-dimethylaminomethylene)amine, N,N′-isopropylidenediamine, N-p-nitrobenzylideneamine, N-salicylideneamine, N-5-chlorosalicylideneamine, N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine, N-cyclohexylideneamine, N-(5,5-dimethyl-3-oxo-1-cyclohexenyl)amine, N-borane derivative, N-diphenylborinic acid derivative, N-[phenyl(pentaacylchromium- or tungsten)acyl]amine, N-copper chelate, N-zinc chelate, N-nitroamine, N-nitrosoamine, amine N-oxide, diphenylphosphinamide (Dpp), dimethylthiophosphinamide (Mpt), diphenylthiophosphinamide (Ppt), dialkyl phosphoramidates, dibenzyl phosphoramidate, diphenyl phosphoramidate, benzenesulfenamide, o-nitrobenzenesulfenamide (Nps), 2,4-dinitrobenzenesulfenamide, pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide, triphenylmethylsulfenamide, and 3-nitropyridinesulfenamide (Npys).

In certain embodiments, the substituent present on an oxygen atom is an oxygen protecting group (also referred to as a hydroxyl protecting group). Oxygen protecting groups include, but are not limited to, —Raa, —N(Rbb)2, —C(═O)SRaa, —C(═O)Raa, —CO2Raa, —C(═O)N(Rbb)2, —C(═NRbb)Raa, —C(═NRbb)ORaa, —C(═NRbb)N(Rbb)2, —S(═O)Raa, —SO2Raa, Si(Raa)3, —P(Rcc)2, —P(Rcc)3, —P(═O)2Raa, —P(═O)(Raa)2, —P(═O)(ORcc)2, —P(═O)2N(Rbb)2, and —P(═O)(NRbb)2, wherein Raa, Rbb, and Rcc are as defined herein. Oxygen protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis , T. W. Greene and P. G. M. Wuts, 3rd edition, John Wiley & Sons, 1999, incorporated herein by reference.

Exemplary oxygen protecting groups include, but are not limited to, methyl, methoxylmethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl, (phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM), p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM), siloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR), tetrahydropyranyl (THP), 3-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl (MTHP), 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl S,S-dioxide, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl (CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl, 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl, 1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl, 2,2,2-trichloroethyl, 2-trimethyl silylethyl, 2-(phenylselenyl)ethyl, t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, benzyl (Bn), p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2-picolyl, 4-picolyl, 3-methyl-2-picoly1 N-oxido, diphenylmethyl, p,p′-dinitrobenzhydryl, 5-dibenzosuberyl, triphenylmethyl, α-naphthyldiphenylmethyl, p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl, tri(p-methoxyphenyl)methyl, 4-(4′-bromophenacyloxyphenyl)diphenylmethyl, 4,4′,4″-tris(4,5-dichlorophthalimidophenyl)methyl, 4,4′,4″-tris(levulinoyloxyphenyl)methyl, 4,4′,4″-tris(benzoyloxyphenyl)methyl, 3-(imidazol-1-yl)bis(4′,4″-dimethoxyphenyl)methyl, 1,1-bis(4-methoxyphenyl)-1′-pyrenylmethyl, 9-anthryl, 9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl, 1,3-benzodisulfuran-2-yl, benzisothiazolyl S,S-dioxido, trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS), dimethylthexylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsily1 (TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TBMPS), formate, benzoylformate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate (levulinate), 4,4-(ethylenedithio)pentanoate (levulinoyldithioacetal), pivaloate, adamantoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate, 2,4,6-trimethylbenzoate (mesitoate), alkyl methyl carbonate, 9-fluorenylmethyl carbonate (Fmoc), alkyl ethyl carbonate, alkyl 2,2,2-trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl carbonate (TMSEC), 2-(phenylsulfonyl) ethyl carbonate (Psec), 2-(triphenylphosphonio) ethyl carbonate (Peoc), alkyl isobutyl carbonate, alkyl vinyl carbonate alkyl allyl carbonate, alkyl p-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzyl carbonate, alkyl 3,4-dimethoxybenzyl carbonate, alkyl o-nitrobenzyl carbonate, alkyl p-nitrobenzyl carbonate, alkyl S-benzyl thiocarbonate, 4-ethoxy-1-napththyl carbonate, methyl dithiocarbonate, 2-iodobenzoate, 4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate, 2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate, 2,6-dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate, 2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate, o-(methoxyacyl)benzoate, a-naphthoate, nitrate, alkyl N,N,N′,N′-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate, borate, dimethylphosphinothioyl, alkyl 2,4-dinitrophenylsulfenate, sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate (Ts).

In certain embodiments, the substituent present on an sulfur atom is an sulfur protecting group (also referred to as a thiol protecting group). Sulfur protecting groups include, but are not limited to, —Raa, —N(Rbb)2, —C(═O)SRaa, —C(═O)Raa, —CO2Raa, C(═O)N(Rbb)2, —C(═NRbb)Raa, —C(═NRbb)ORaa, —C(═NRbb)N(Rbb)2, —S(═O)Raa, —SO2Raa, —Si(Raa)3, —P(Rcc)2, —P(Rcc)3, —P(═O)2Raa, —P(═O)(Raa)2, —P(═O)(ORcc)2, —P(═O)2N(Rbb)2, and —P(═O)(NRbb)2, wherein Raa, Rbb, and Rcc are as defined herein. Sulfur protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis , T. W. Greene and P. G. M. Wuts, 3rd edition, John Wiley & Sons, 1999, incorporated herein by reference.

As used herein, a “leaving group” is an art-understood term referring to a molecular fragment that departs with a pair of electrons in heterolytic bond cleavage, wherein the molecular fragment is an anion or neutral molecule. See, for example, Smith, March's Advanced Organic Chemistry 6th ed. (501-502). Exemplary leaving groups include, but are not limited to, halo (e.g., chloro, bromo, iodo) and sulfonyl substituted hydroxyl groups (e.g., tosyl, mesyl, besyl).

Other Definitions

As used herein, use of the phrase “at least one instance” refers to one instance, but also encompasses more than one instance, e.g., for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 instances, and up to 100 instances.

As used herein, a “polymer” refers to a compound comprised of at least 3 (e.g., at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, etc.) repeating covalently bound structural units.

“Attached” refers to the covalent attachment of a group.

As used herein, “lipophilic” refers to the ability of a group to dissolve in fats, oils, lipids, and lipophilic non-polar solvents such as hexane or toluene. In general, a lipophilic group refers to an unsubstituted n-alkyl or unsubstituted n-alkenyl group having 6 to 50 carbon atoms, e.g., 6 to 40, 6 to 30, 6 to 20, 8 to 20, 8 to 19, 8 to 18, 8 to 17, 8 to 16, or 8 to 15 carbon atoms.

As used herein, the term “salt” or “pharmaceutically acceptable salt” refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al., describes pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66:1-19. Pharmaceutically acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or rnalonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate. digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like. Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N+(C1-4alkyl)4 salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium. quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, sulfonate and aryl sulfonate. Further pharmaceutically acceptable salts include salts formed from the quarternization of an amine using an appropriate electrophile, e.g., an alkyl halide, to form a quarternized alkylated amino salt.

Second or Additional Cationic Lipids

In some embodiments, liposomes may comprise a second or additional cationic lipid. As used herein, the phrase “cationic lipid” refers to any of a number of lipid species that have a net positive charge at a selected pH, such as physiological pH. Several cationic lipids have been described in the literature, many of which are commercially available. Particularly suitable cationic lipids for use in the compositions and methods of the invention include those described in international patent publications WO 2010/053572 (and particularly, C12-200 described at paragraph [00225]) and WO 2012/170930, both of which are incorporated herein by reference. In certain embodiments, the compositions and methods of the invention employ a lipid nanoparticles comprising an ionizable cationic lipid described in U.S. provisional patent application 61/617,468, filed Mar. 29, 2012 (incorporated herein by reference), such as, e.g., (15Z,18Z)—N,N-dimethyl-6-(9Z,12Z)-octadeca-9, 12-dien-1-yl)tetracosa-15,18-dien-1-amine (HGT5000), (15Z,18Z)—N,N-dimethyl-6-((9Z,12Z)-octadeca-9, 12-dien-1-yl)tetracosa-4,15,18-trien-1-amine (HGT5001), and (15Z,18Z)—N,N-dimethyl-6-((9Z,12Z)-octadeca-9, 12-dien-1-yl)tetracosa-5,15,18-trien-1-amine (HGT5002).

In some embodiments, the second or additional cationic lipid N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride or “DOTMA” is used. (Feigner et al. (Proc. Nat'l Acad. Sci. 84, 7413 (1987); U.S. Pat. No. 4,897,355). DOTMA can be formulated alone or can be combined with the neutral lipid, dioleoylphosphatidyl-ethanolamine or “DOPE” or other cationic or non-cationic lipids into a liposomal transfer vehicle or a lipid nanoparticle, and such liposomes can be used to enhance the delivery of nucleic acids into target cells. Other suitable cationic lipids include, for example, 5-carboxyspermylglycinedioctadecylamide or “DOGS,” 2,3-dioleyloxy-N-[2(spermine-carboxamido)ethyl]-N,N-dimethyl-1-propanaminium or “DOSPA” (Behr et al. Proc. Nat.'l Acad. Sci. 86, 6982 (1989); U.S. Pat. Nos. 5,171,678; 5,334,761), 1,2-Dioleoyl-3-Dimethylammonium-Propane or “DODAP”, 1,2-Dioleoyl-3-Trimethylammonium-Propane or “DOTAP”. Additional exemplary cationic lipids also include 1,2-distearyloxy-N,N-dimethyl-3-aminopropane or “DSDMA”, 1,2-dioleyloxy-N,N-dimethyl-3-aminopropane or “DODMA”, 1,2-dilinoleyloxy-N,N-dimethyl-3-aminopropane or “DLinDMA”, 1,2-dilinolenyloxy-N,N-dimethyl-3-aminopropane or “DLenDMA”, N-dioleyl-N,N-dimethylammonium chloride or “DODAC”, N,N-distearyl-N,N-dimethylarnrnonium bromide or “DDAB”, N-(1,2-dimyristyloxyprop-3-yl)-N,N-dimethyl-N-hydroxyethyl ammonium bromide or “DMRIE”, 3-dimethylamino-2-(cholest-5-en-3-beta-oxybutan-4-oxy)-1-(cis,cis-9,12-octadecadienoxy)propane or “CLinDMA”, 2-[5′-(cholest-5-en-3-beta-oxy)-3′-oxapentoxy)-3-dimethy 1-1-(cis,cis-9′,1-2′-octadecadienoxy)propane or “CpLinDMA”, N,N-dimethyl-3,4-dioleyloxybenzylamine or “DMOBA”, 1,2-N,N′-dioleylcarbamyl-3-dimethylaminopropane or “DOcarbDAP”, 2,3-Dilinoleoyloxy-N,N-dimethylpropylamine or “DLinDAP”, 1,2-N,N′-Dilinoleylcarbamyl-3-dimethylaminopropane or “DLincarbDAP”, 1,2-Dilinoleoylcarbamyl-3-dimethylaminopropane or “DLinCDAP”, 2,2-dilinoleyl-4-dimethylaminomethyl-[1,3]-dioxolane or “DLin- -DMA”, 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane or “DLin-K-XTC2-DMA”, and 2-(2,2-di((9Z,12Z)-octadeca-9,12-dien-1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylethanamine (DLin-KC2-DMA)) (See, WO 2010/042877; Semple et al., Nature Biotech. 28: 172-176 (2010)), or mixtures thereof. (Heyes, J., et al., J Controlled Release 107: 276-287 (2005); Morrissey, D V., et al., Nat. Biotechnol. 23(8): 1003-1007 (2005); PCT Publication WO2005/121348A1). In some embodiments, one or more of the cationic lipids comprise at least one of an imidazole, dialkylamino, or guanidinium moiety.

In some embodiments, the second or additional cationic lipid may be chosen from XTC (2,2-Dilinoley 1-4-dimethylaminoethy 1-[1,3]-dioxolane), MC3 (((6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl 4-(dimethylamino)butanoate), ALNY-100 ((3 aR,5 s,6aS)—N,N-dimethyl-2,2-di((9Z,12Z)-octadeca-9,12-dienyl)tetrahydro-3aH-cyclopenta[d][1,3]dioxol-5-amine)), NC98-5 (4,7,13-tris(3-oxo-3-(undecylamino)propyl)-N1,N16-diundecyl-4,7,10,13-tetraazahexadecane-1,16-diamide), DODAP (1,2-dioleyl-3-dimethylammonium propane), HGT4003 (WO 2012/170889, the teachings of which are incorporated herein by reference in their entirety), ICE (WO 2011/068810, the teachings of which are incorporated herein by reference in their entirety), HGT5000 (U.S. Provisional Patent Application No. 61/617,468, the teachings of which are incorporated herein by reference in their entirety) or HGT5001 (cis or trans) (Provisional Patent Application No. 61/617,468), aminoalcohol lipidoids such as those disclosed in WO2010/053572, DOTAP (1,2-dioleyl-3-trimethylammonium propane), DOTMA (1,2-di-O-octadecenyl-3-trimethylammonium propane), DLinDMA (Heyes, J.; Palmer, L.; Bremner, K.; MacLachlan, I. “Cationic lipid saturation influences intracellular delivery of encapsulated nucleic acids” J. Contr. Rel. 2005, 107, 276-287), DLin-KC2-DMA (Semple, S. C. et al. “Rational Design of Cationic Lipids for siRNA Delivery” Nature Biotech. 2010, 28, 172-176), C12-200 (Love, K. T. et al. “Lipid-like materials for low-dose in vivo gene silencing” PNAS 2010, 107, 1864-1869).

Non-Cationic/Helper Lipids

In some embodiments, provided liposomes contain one or more non-cationic (“helper”) lipids. As used herein, the phrase “non-cationic lipid” refers to any neutral, zwitterionic or anionic lipid. As used herein, the phrase “anionic lipid” refers to any of a number of lipid species that carry a net negative charge at a selected H, such as physiological pH. Non-cationic lipids include, but are not limited to, distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoyl-phosphatidylethanolamine (POPE), dioleoyl-phosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl-phosphatidyl-ethanolamine (DSPE), 16-O-monomethyl PE, 16-O-dimethyl PE, 18-1-trans PE, 1-stearoyl-2-oleoyl-phosphatidyethanolamine (SOPE), or a mixture thereof.

In some embodiments, such non-cationic lipids may be used alone, but are preferably used in combination with other excipients, for example, cationic lipids. In some embodiments, the non-cationic lipid may comprise a molar ratio of about 5% to about 90%, or about 10% to about 70% of the total lipid present in a liposome. In some embodiments, a non-cationic lipid is a neutral lipid, i.e., a lipid that does not carry a net charge in the conditions under which the composition is formulated and/or administered. In some embodiments, the percentage of non-cationic lipid in a liposome may be greater than 5%, greater than 10%, greater than 20%, greater than 30%, or greater than 40%.

Cholesterol-Based Lipids

In some embodiments, provided liposomes comprise one or more cholesterol-based lipids. For example, suitable cholesterol-based cationic lipids include, for example, DC-Choi (N,N-dimethyl-N-ethylcarboxamidocholesterol), 1,4-bis(3-N-oleylamino-propyl)piperazine (Gao, et al. Biochem. Biophys. Res. Comm. 179, 280 (1991); Wolf et al. BioTechniques 23, 139 (1997); U.S. Pat. No. 5,744,335), or ICE. In some embodiments, the cholesterol-based lipid may comprise a molar ration of about 2% to about 30%, or about 5% to about 20% of the total lipid present in a liposome. In some embodiments, The percentage of cholesterol-based lipid in the lipid nanoparticle may be greater than 5, %, 10%, greater than 20%, greater than 30%, or greater than 40%.

PEGylated Lipids

In some embodiments, provided liposomes comprise one or more PEGylated lipids. For example, the use of polyethylene glycol (PEG)-modified phospholipids and derivatized lipids such as derivatized ceramides (PEG-CER), including N-Octanoyl-Sphingosine-1-[Succinyl(Methoxy Polyethylene Glycol)-2000] (C8 PEG-2000 ceramide) is also contemplated by the present invention in combination with one or more of the cationic and, in some embodiments, other lipids together which comprise the liposome. Contemplated PEG-modified lipids include, but are not limited to, a polyethylene glycol chain of up to 5 kDa in length covalently attached to a lipid with alkyl chain(s) of C 6 -C 20 length. In some embodiments, a PEG-modified or PEGylated lipid is PEGylated cholesterol or PEG-2K. The addition of such components may prevent complex aggregation and may also provide a means for increasing circulation lifetime and increasing the delivery of the lipid-nucleic acid composition to the target cell, (Klibanov et al. (1990) FEBS Letters, 268 (1): 235-237), or they may be selected to rapidly exchange out of the formulation in vivo (see U.S. Pat. No. 5,885,613).

In some embodiments, particularly useful exchangeable lipids are PEG-ceramides having shorter acyl chains (e.g., C 14 or C 18 ). The PEG-modified phospholipid and derivitized lipids of the present invention may comprise a molar ratio from about 0% to about 15%, about 0.5% to about 15%, about 1% to about 15%, about 4% to about 10%, or about 2% of the total lipid present in the liposome.

According to various embodiments, the selection of second or additional cationic lipids, non-cationic lipids and/or PEG-modified lipids which comprise the lipid nanoparticle, as well as the relative molar ratio of such lipids to each other, is based upon the characteristics of the selected lipid(s), the nature of the intended target cells, the characteristics of the mRNA to be delivered. Additional considerations include, for example, the saturation of the alkyl chain, as well as the size, charge, pH, pKa, fusogenicity and toxicity of the selected lipid(s). Thus the molar ratios may be adjusted accordingly. In some embodiments, the percentage of PEG-modified lipid in a liposome may be greater than 1%, greater than 2%, greater than 5%, greater than 10%, or greater than 15%.

Polymer

In some embodiments, a suitable liposome according to the present invention further includes a polymer, in combination with one or more cationic lipids as described and, in some embodiments, other carriers including various lipids described herein. Thus, in some embodiments, liposomal delivery vehicles, as used herein, also encompass polymer containing nanoparticles. Suitable polymers may include, for example, polyacrylates, polyalkycyanoacrylates, polylactide, polylactide-polyglycolide copolymers, polycaprolactones, dextran, albumin, gelatin, alginate, collagen, chitosan, cyclodextrins, protamine, PEGylated protamine, PLL, PEGylated PLL and polyethylenimine (PEI). When PEI is present, it may be branched PEI of a molecular weight ranging from 10 to 40 kDA, e.g., 25 kDa branched PEI (Sigma #408727).

In some embodiments, a suitable liposome formulation contains a combination of one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids one or more PEG-modified lipids, and/or one or more polymers. As a non-limiting example, a suitable liposome comprises cKK-E12, DOPE, cholesterol and DMG-PEG2K. In some embodiments, the ratio of cationic lipid to non-cationic lipid to cholesterol-based lipid to PEGylated lipid may be between about 30-50:25-35:20-30:1-15, respectively. In some embodiments, the ratio of cationic lipid to non-cationic lipid to cholesterol-based lipid to PEGylated lipid is approximately 40:30:20:10, respectively. In some embodiments, the ratio of cationic lipid to non-cationic lipid to cholesterol-based lipid to PEGylated lipid is approximately 40:30:25:5, respectively. In some embodiments, the ratio of cationic lipid to non-cationic lipid to cholesterol-based lipid to PEGylated lipid is approximately 40:32:25:3, respectively.

mRNA

The present invention can be used to deliver any mRNA. mRNA is typically thought of as the type of RNA that carries information from DNA to the ribosome. The existence of mRNA is usually very brief and includes processing and translation, followed by degradation. Typically, in eukaryotic organisms, mRNA processing comprises the addition of a “cap” on the N-terminal (5′) end, and a “tail” on the C-terminal (3′) end. A typical cap is a 7-methylguanosine cap, which is a guanosine that is linked through a 5′-5′-triphosphate bond to the first transcribed nucleotide. The presence of the cap is important in providing resistance to nucleases found in most eukaryotic cells. The tail is typically a polyadenylation event whereby a polyadenylyl moiety is added to the 3′ end of the mRNA molecule. The presence of this “tail” serves to protect the mRNA from exonuclease degradation. Messenger RNA typically is translated by the ribosomes into a series of amino acids that make up a protein.

Any mRNA capable of being translated into one or more peptides (e.g., proteins) or peptide fragments is contemplated as within the scope of the present invention. In some embodiments, an mRNA encodes one or more naturally occurring peptides. In some embodiments, an mRNA encodes one or more modified or non-natural peptides.

In some embodiments an mRNA encodes an intracellular protein. In some embodiments, an mRNA encodes a cytosolic protein. In some embodiments, an mRNA encodes a protein associated with the actin cytoskeleton. In some embodiments, an mRNA encodes a protein associated with the plasma membrane. In some specific embodiments, an mRNA encodes a transmembrane protein. In some specific embodiments an mRNA encodes an ion channel protein. In some embodiments, an mRNA encodes a perinuclear protein. In some embodiments, an mRNA encodes a nuclear protein. In some specific embodiments, an mRNA encodes a transcription factor. In some embodiments, an mRNA encodes a chaperone protein. In some embodiments, an mRNA encodes an intracellular enzyme (e.g., mRNA encoding an enzyme associated with urea cycle or lysosomal storage metabolic disorders). In some embodiments, an mRNA encodes a protein involved in cellular metabolism, DNA repair, transcription and/or translation. In some embodiments, an mRNA encodes an extracellular protein. In some embodiments, an mRNA encodes a protein associated with the extracellular matrix. In some embodiments an mRNA encodes a secreted protein. In specific embodiments, an mRNA used in the composition and methods of the invention may be used to express functional proteins or enzymes that are excreted or secreted by one or more target cells into the surrounding extracellular fluid (e.g., mRNA encoding hormones and/or neurotransmitters).

In some embodiments, the compositions and methods of the invention provide for delivery of mRNA encoding a secreted protein. In some embodiments, the compositions and methods of the invention provide for delivery of mRNA encoding one or more secreted proteins listed in Table 1; thus, compositions of the invention may comprise an mRNA encoding a protein listed in Table 1 (or a homolog thereof) along with other components set out herein, and methods of the invention may comprise preparing and/or administering a composition comprising an mRNA encoding a protein listed in Table 1 (or a homolog thereof) along with other components set out herein.

In some embodiments, the compositions and methods of the invention provide for the delivery of one or more mRNAs encoding one or more additional exemplary proteins listed in Table 2; thus, compositions of the invention may comprise an mRNA encoding a protein listed in Table 2 (or a homolog thereof) along with other components set out herein, and methods of the invention may comprise preparing and/or administering a composition comprising an mRNA encoding a protein chosen from the proteins listed in Table 2 (or a homolog thereof) along with other components set out herein.

The Uniprot IDs set forth in Table 1 and Table 2 refer to the human versions the listed proteins and the sequences of each are available from the Uniprot database. Sequences of the listed proteins are also generally available for various animals, including various mammals and animals of veterinary or industrial interest. Accordingly, in some embodiments, compositions and methods of the invention provide for the delivery of one or more mRNAs encoding one or more proteins chosen from mammalian homologs or homologs from an animal of veterinary or industrial interest of the secreted proteins listed in Table 1 or Table 2; thus, compositions of the invention may comprise an mRNA encoding a protein chosen from mammalian homologs or homologs from an animal of veterinary or industrial interest of a protein listed in Table 1 or Table 2 along with other components set out herein, and methods of the invention may comprise preparing and/or administering a composition comprising an mRNA encoding a protein chosen from mammalian homologs or homologs from an animal of veterinary or industrial interest of a protein listed in Table 1 or Table 2 along with other components set out herein. In some embodiments, mammalian homologs are chosen from mouse, rat, hamster, gerbil, horse, pig, cow, llama, alpaca, mink, dog, cat, ferret, sheep, goat, or camel homologs. In some embodiments, the animal of veterinary or industrial interest is chosen from the mammals listed above and/or chicken, duck, turkey, salmon, catfish, or tilapia.

In embodiments, the compositions and methods of the invention provide for the delivery of mRNA encoding a lysosomal protein chosen from Table 3. In some embodiments, the compositions and methods of the invention provide for the delivery of one or more mRNAs encoding one or more lysosomal and/or related proteins listed in Table 3; thus, compositions of the invention may comprise an mRNA encoding a protein listed in Table 3 (or a homolog thereof) along with other components set out herein, and methods of the invention may comprise preparing and/or administering a composition comprising an mRNA encoding a protein chosen from the proteins listed in Table 3 (or a homolog thereof) along with other components set out herein.

Information regarding lysosomal proteins is available from Lubke et al., “Proteomics of the Lysosome,” Biochim Biophys Acta . (2009) 1793: 625-635. In some embodiments, the protein listed in Table 3 and encoded by mRNA in the compositions and methods of the invention is a human protein. Sequences of the listed proteins are also available for various animals, including various mammals and animals of veterinary or industrial interest as described above.

In some embodiments, the compositions and methods of the invention provide for the delivery of mRNA encoding a therapeutic protein (e.g., cytosolic, transmembrane or secreted) such as those listed in Table 4. In some embodiments, the compositions and methods of the invention provide for the delivery of an mRNA encoding a therapeutic protein useful in treating a disease or disorder (i.e., indication) listed in Table 4; thus, compositions of the invention may comprise an mRNA encoding a therapeutic protein listed or not listed in Table 4 (or a homolog thereof, as discussed below) along with other components set out herein for treating a disease or disorder (i.e., indication) listed in Table 4, and methods of the invention may comprise preparing and/or administering a composition comprising an mRNA encoding a such a protein (or a homolog thereof, as discussed below) along with other components set out herein for treatment of a disease or disorder listed in Table 4.

In some embodiments, the present invention is used to prevent, treat and/or cure a subject affected with a disease or disorder listed or associated with the proteins listed in Tables 1, 2, 3 or 4. In some embodiments, an mRNA encodes one or more of argininosuccinate synthetase (ASS1), Factor IX, survival motor neuron 1 (SMN1), or phenylalanine hydroxylase

Synthesis of mRNA

mRNAs according to the present invention may be synthesized according to any of a variety of known methods. For example, mRNAs according to the present invention may be synthesized via in vitro transcription (IVT). Briefly, IVT is typically performed with a linear or circular DNA template containing a promoter, a pool of ribonucleotide triphosphates, a buffer system that may include DTT and magnesium ions, and an appropriate RNA polymerase (e.g., T3, T7 or SP6 RNA polymerase), DNAse I, pyrophosphatase, and/or RNAse inhibitor. The exact conditions will vary according to the specific application.

In some embodiments, for the preparation of mRNA according to the invention, a DNA template is transcribed in vitro. A suitable DNA template typically has a promoter, for example a T3, T7 or SP6 promoter, for in vitro transcription, followed by desired nucleotide sequence for desired mRNA and a termination signal.

Desired mRNA sequence(s) according to the invention may be determined and incorporated into a DNA template using standard methods. For example, starting from a desired amino acid sequence (e.g., an enzyme sequence), a virtual reverse translation is carried out based on the degenerated genetic code. Optimization algorithms may then be used for selection of suitable codons. Typically, the G/C content can be optimized to achieve the highest possible G/C content on one hand, taking into the best possible account the frequency of the tRNAs according to codon usage on the other hand. The optimized RNA sequence can be established and displayed, for example, with the aid of an appropriate display device and compared with the original (wild-type) sequence. A secondary structure can also be analyzed to calculate stabilizing and destabilizing properties or, respectively, regions of the RNA.

Modified mRNA

In some embodiments, mRNA according to the present invention may be synthesized as unmodified or modified mRNA. Typically, mRNAs are modified to enhance stability. Modifications of mRNA can include, for example, modifications of the nucleotides of the RNA. An modified mRNA according to the invention can thus include, for example, backbone modifications, sugar modifications or base modifications. In some embodiments, mRNAs may be synthesized from naturally occurring nucleotides and/or nucleotide analogues (modified nucleotides) including, but not limited to, purines (adenine (A), guanine (G)) or pyrimidines (thymine (T), cytosine (C), uracil (U)), and as modified nucleotides analogues or derivatives of purines and pyrimidines, such as e.g. 1-methyl-adenine, 2-methyl-adenine, 2-methylthio-N-6-isopentenyl-adenine, N6-methyl-adenine, N6-isopentenyl-adenine, 2-thio-cytosine, 3-methyl-cytosine, 4-acetyl-cytosine, 5-methyl-cytosine, 2,6-diaminopurine, 1-methyl-guanine, 2-methyl-guanine, 2,2-dimethyl-guanine, 7-methyl-guanine, inosine, 1-methyl-inosine, pseudouracil (5-uracil), dihydro-uracil, 2-thio-uracil, 4-thio-uracil, 5-carboxymethylaminomethyl-2-thio-uracil, 5-(carboxyhydroxymethyl)-uracil, 5-fluoro-uracil, 5-bromo-uracil, 5-carboxymethylaminomethyl-uracil, 5-methyl-2-thio-uracil, 5-methyl-uracil, N-uracil-5-oxyacetic acid methyl ester, 5-methylaminomethyl-uracil, 5-methoxyaminomethyl-2-thio-uracil, 5′-methoxycarbonylmethyl-uracil, 5-methoxy-uracil, uracil-5-oxyacetic acid methyl ester, uracil-5-oxyacetic acid (v), 1-methyl-pseudouracil, queosine, .beta.-D-mannosyl-queosine, wybutoxosine, and phosphoramidates, phosphorothioates, peptide nucleotides, methylphosphonates, 7-deazaguanosine, 5-methylcytosine and inosine. The preparation of such analogues is known to a person skilled in the art e.g. from the U.S. Pat. Nos. 4,373,071, 4,401,796, 4,415,732, 4,458,066, 4,500,707, 4,668,777, 4,973,679, 5,047,524, 5,132,418, 5,153,319, 5,262,530 and 5,700,642, the disclosures of which are incorporated by reference in their entirety.

In some embodiments, mRNAs (e.g., enzyme encoding mRNAs) may contain RNA backbone modifications. Typically, a backbone modification is a modification in which the phosphates of the backbone of the nucleotides contained in the RNA are modified chemically. Exemplary backbone modifications typically include, but are not limited to, modifications from the group consisting of methylphosphonates, methylphosphoramidates, phosphoramidates, phosphorothioates (e.g. cytidine 5′-O-(1-thiophosphate)), boranophosphates, positively charged guanidinium groups etc., which means by replacing the phosphodiester linkage by other anionic, cationic or neutral groups.

In some embodiments, mRNAs (e.g., enzyme encoding mRNAs) may contain sugar modifications. A typical sugar modification is a chemical modification of the sugar of the nucleotides it contains including, but not limited to, sugar modifications chosen from the group consisting of 2′-deoxy-2′-fluoro-oligoribonucleotide (2′-fluoro-2′-deoxycytidine 5′-triphosphate, 2′-fluoro-2′-deoxyuridine 5′-triphosphate), 2′-deoxy-2′-deamine-oligoribonucleotide (2′-amino-2′-deoxycytidine 5′-triphosphate, 2′-amino-2′-deoxyuridine 5′-triphosphate), 2′-O-alkyloligoribonucleotide, 2′-deoxy-2′-C-alkyloligoribonucleotide (2′-O-methylcytidine 5′-triphosphate, 2′-methyluridine 5′-triphosphate), 2′-C-alkyloligoribonucleotide, and isomers thereof (2′-aracytidine 5′-triphosphate, 2′-arauridine 5′-triphosphate), or azidotriphosphates (2′-azido-2′-deoxycytidine 5′-triphosphate, 2′-azido-2′-deoxyuridine 5′-triphosphate).

In some embodiments, mRNAs (e.g., enzyme encoding mRNAs) may contain modifications of the bases of the nucleotides (base modifications). A modified nucleotide which contains a base modification is also called a base-modified nucleotide. Examples of such base-modified nucleotides include, but are not limited to, 2-amino-6-chloropurine riboside 5′-triphosphate, 2-aminoadenosine 5′-triphosphate, 2-thiocytidine 5′-triphosphate, 2-thiouridine 5′-triphosphate, 4-thiouridine 5′-triphosphate, 5-aminoallylcytidine 5′-triphosphate, 5-aminoallyluridine 5′-triphosphate, 5-bromocytidine 5′-triphosphate, 5-bromouridine 5′-triphosphate, 5-iodocytidine 5′-triphosphate, 5-iodouridine 5′-triphosphate, 5-methylcytidine 5′-triphosphate, 5-methyluridine 5′-triphosphate, 6-azacytidine 5′-triphosphate, 6-azauridine 5′-triphosphate, 6-chloropurine riboside 5′-triphosphate, 7-deazaadenosine 5′-triphosphate, 7-deazaguanosine 5′-triphosphate, 8-azaadenosine 5′-triphosphate, 8-azidoadenosine 5′-triphosphate, benzimidazole riboside 5′-triphosphate, N1-methyladenosine 5′-triphosphate, N1-methylguanosine 5′-triphosphate, N6-methyladenosine 5′-triphosphate, 06-methylguanosine 5′-triphosphate, pseudouridine 5′-triphosphate, puromycin 5′-triphosphate or xanthosine 5′-triphosphate.

Cap Structure

Typically, mRNA synthesis includes the addition of a “cap” on the N-terminal (5′) end, and a “tail” on the C-terminal (3′) end. The presence of the cap is important in providing resistance to nucleases found in most eukaryotic cells. The presence of a “tail” serves to protect the mRNA from exonuclease degradation.

Thus, in some embodiments, mRNAs (e.g., enzyme encoding mRNAs) include a 5′ cap structure. A 5′ cap is typically added as follows: first, an RNA terminal phosphatase removes one of the terminal phosphate groups from the 5′ nucleotide, leaving two terminal phosphates; guanosine triphosphate (GTP) is then added to the terminal phosphates via a guanylyl transferase, producing a 5′5′5 triphosphate linkage; and the 7-nitrogen of guanine is then methylated by a methyltransferase. Examples of cap structures include, but are not limited to, m7G(5′)ppp (5′(A,G(5′)ppp(5′)A and G(5′)ppp(5′)G.

In some embodiments, naturally occurring cap structures comprise a 7-methyl guanosine that is linked via a triphosphate bridge to the 5′-end of the first transcribed nucleotide, resulting in a dinucleotide cap of m 7 G(5′)ppp(5′)N, where N is any nucleoside. In vivo, the cap is added enzymatically. The cap is added in the nucleus and is catalyzed by the enzyme guanylyl transferase. The addition of the cap to the 5′ terminal end of RNA occurs immediately after initiation of transcription. The terminal nucleoside is typically a guanosine, and is in the reverse orientation to all the other nucleotides, i.e., G(5′)ppp(5′)GpNpNp.

A common cap for mRNA produced by in vitro transcription is m 7 G(5′)ppp(5′)G, which has been used as the dinucleotide cap in transcription with T7 or SP6 RNA polymerase in vitro to obtain RNAs having a cap structure in their 5′-termini. The prevailing method for the in vitro synthesis of capped mRNA employs a pre-formed dinucleotide of the form m 7 G(5′)ppp(5′)G (“m 7 GpppG”) as an initiator of transcription.

To date, a usual form of a synthetic dinucleotide cap used in in vitro translation experiments is the Anti-Reverse Cap Analog (“ARCA”) or modified ARCA, which is generally a modified cap analog in which the 2′ or 3′ OH group is replaced with —OCH 3 .

Additional cap analogs include, but are not limited to, chemical structures selected from the group consisting of m 7 GpppG, m 7 GpppA, m 7 GpppC; unmethylated cap analogs (e.g., GpppG); dimethylated cap analog (e.g., m 2 ′7GpppG), trimethylated cap analog (e.g., m 2,2,7 GpppG), dimethylated symmetrical cap analogs (e.g., m 7 Gpppm 7 G), or anti reverse cap analogs (e.g., ARCA; m 7 , 2′Ome GpppG, m 72′d GpppG, m 7,3′Ome GpppG, m 7,3′d GpppG and their tetraphosphate derivatives) (see, e.g., Jemielity, J. et al., “ Novel ‘anti - reverse’ cap analogs with superior translational properties ”, RNA, 9: 1108-1122 (2003)).

In some embodiments, a suitable cap is a 7-methyl guanylate (“m 7 G”) linked via a triphosphate bridge to the 5′-end of the first transcribed nucleotide, resulting in m 7 G(5′)ppp(5′)N, where N is any nucleoside. A preferred embodiment of a m 7 G cap utilized in embodiments of the invention is m 7 G(5′)ppp(5′)G.

In some embodiments, the cap is a Cap0 structure. Cap0 structures lack a 2′-O-methyl residue of the ribose attached to bases 1 and 2. In some embodiments, the cap is a Cap1 structure. Cap1 structures have a 2′-O-methyl residue at base 2. In some embodiments, the cap is a Cap2 structure. Cap2 structures have a 2′-O-methyl residue attached to both bases 2 and 3.

A variety of m 7 G cap analogs are known in the art, many of which are commercially available. These include the m 7 GpppG described above, as well as the ARCA 3′-OCH 3 and 2′-OCH 3 cap analogs (Jemielity, J. et al., RNA, 9: 1108-1122 (2003)). Additional cap analogs for use in embodiments of the invention include N7-benzylated dinucleoside tetraphosphate analogs (described in Grudzien, E. et al., RNA, 10: 1479-1487 (2004)), phosphorothioate cap analogs (described in Grudzien-Nogalska, E., et al., RNA, 13: 1745-1755 (2007)), and cap analogs (including biotinylated cap analogs) described in U.S. Pat. Nos. 8,093,367 and 8,304,529, incorporated by reference herein.

Tail Structure

Typically, the presence of a “tail” serves to protect the mRNA from exonuclease degradation. The poly A tail is thought to stabilize natural messengers and synthetic sense RNA. Therefore, in certain embodiments a long poly A tail can be added to an mRNA molecule thus rendering the RNA more stable. Poly A tails can be added using a variety of art-recognized techniques. For example, long poly A tails can be added to synthetic or in vitro transcribed RNA using poly A polymerase (Yokoe, et al. Nature Biotechnology. 1996; 14: 1252-1256). A transcription vector can also encode long poly A tails. In addition, poly A tails can be added by transcription directly from PCR products. Poly A may also be ligated to the 3′ end of a sense RNA with RNA ligase (see, e.g., Molecular Cloning A Laboratory Manual, 2nd Ed., ed. by Sambrook, Fritsch and Maniatis (Cold Spring Harbor Laboratory Press: 1991 edition)).

In some embodiments, mRNAs (e.g., enzyme encoding mRNAs) include a 3′ poly(A) tail structure. Typically, the length of the poly A tail can be at least about 10, 50, 100, 200, 300, 400 at least 500 nucleotides (SEQ ID NO: 12). In some embodiments, a poly-A tail on the 3′ terminus of mRNA typically includes about 10 to 300 adenosine nucleotides (SEQ ID NO: 13) (e.g., about 10 to 200 adenosine nucleotides, about 10 to 150 adenosine nucleotides, about 10 to 100 adenosine nucleotides, about 20 to 70 adenosine nucleotides, or about 20 to 60 adenosine nucleotides). In some embodiments, mRNAs include a 3′ poly(C) tail structure. A suitable poly-C tail on the 3′ terminus of mRNA typically include about 10 to 200 cytosine nucleotides (SEQ ID NO: 14) (e.g., about 10 to 150 cytosine nucleotides, about 10 to 100 cytosine nucleotides, about 20 to 70 cytosine nucleotides, about 20 to 60 cytosine nucleotides, or about 10 to 40 cytosine nucleotides). The poly-C tail may be added to the poly-A tail or may substitute the poly-A tail.

In some embodiments, the length of the poly A or poly C tail is adjusted to control the stability of a modified sense mRNA molecule of the invention and, thus, the transcription of protein. For example, since the length of the poly A tail can influence the half-life of a sense mRNA molecule, the length of the poly A tail can be adjusted to modify the level of resistance of the mRNA to nucleases and thereby control the time course of polynucleotide expression and/or polypeptide production in a target cell.

5′ and 3′ Untranslated Region

In some embodiments, mRNAs include a 5′ and/or 3′ untranslated region. In some embodiments, a 5′ untranslated region includes one or more elements that affect an mRNA's stability or translation, for example, an iron responsive element. In some embodiments, a 5′ untranslated region may be between about 50 and 500 nucleotides in length.

In some embodiments, a 3′ untranslated region includes one or more of a polyadenylation signal, a binding site for proteins that affect an mRNA's stability of location in a cell, or one or more binding sites for miRNAs. In some embodiments, a 3′ untranslated region may be between 50 and 500 nucleotides in length or longer.

Exemplary 3′ and/or 5′ UTR sequences can be derived from mRNA molecules which are stable (e.g., globin, actin, GAPDH, tubulin, histone, or citric acid cycle enzymes) to increase the stability of the sense mRNA molecule. For example, a 5′ UTR sequence may include a partial sequence of a CMV immediate-early 1 (IE1) gene, or a fragment thereof to improve the nuclease resistance and/or improve the half-life of the polynucleotide. Also contemplated is the inclusion of a sequence encoding human growth hormone (hGH), or a fragment thereof to the 3′ end or untranslated region of the polynucleotide (e.g., mRNA) to further stabilize the polynucleotide. Generally, these modifications improve the stability and/or pharmacokinetic properties (e.g., half-life) of the polynucleotide relative to their unmodified counterparts, and include, for example modifications made to improve such polynucleotides' resistance to in vivo nuclease digestion.

According to various embodiments, any size mRNA may be encapsulated by provided liposomes. In some embodiments, the provided liposomes may encapsulate mRNA of greater than about 0.5 kb, 1 kb, 1.5 kb, 2 kb, 2.5 kb, 3 kb, 3.5 kb, 4 kb, 4.5 kb, or 5 kb in length.

Formation of Liposomes

The liposomes for use in provided compositions can be prepared by various techniques which are presently known in the art. For example, multilamellar vesicles (MLV) may be prepared according to conventional techniques, such as by depositing a selected lipid on the inside wall of a suitable container or vessel by dissolving the lipid in an appropriate solvent, and then evaporating the solvent to leave a thin film on the inside of the vessel or by spray drying. An aqueous phase may then added to the vessel with a vortexing motion which results in the formation of MLVs. Uni-lamellar vesicles (ULV) can then be formed by homogenization, sonication or extrusion of the multi-lamellar vesicles. In addition, unilamellar vesicles can be formed by detergent removal techniques.

In certain embodiments, provided compositions comprise a liposome wherein the mRNA is associated on both the surface of the liposome and encapsulated within the same liposome. For example, during preparation of the compositions of the present invention, cationic liposomes may associate with the mRNA through electrostatic interactions. For example, during preparation of the compositions of the present invention, cationic liposomes may associate with the mRNA through electrostatic interactions.

In some embodiments, the compositions and methods of the invention comprise mRNA encapsulated in a liposome. In some embodiments, the one or more mRNA species may be encapsulated in the same liposome. In some embodiments, the one or more mRNA species may be encapsulated in different liposomes. In some embodiments, the mRNA is encapsulated in one or more liposomes, which differ in their lipid composition, molar ratio of lipid components, size, charge (Zeta potential), targeting ligands and/or combinations thereof. In some embodiments, the one or more liposome may have a different composition of cationic lipids, neutral lipid, PEG-modified lipid and/or combinations thereof. In some embodiments the one or more lipisomes may have a different molar ratio of cationic lipid, neutral lipid, cholesterol and PEG-modified lipid used to create the liposome.

The process of incorporation of a desired mRNA into a liposome is often referred to as “loading”. Exemplary methods are described in Lasic, et al., FEBS Lett., 312: 255-258, 1992, which is incorporated herein by reference. The liposome-incorporated nucleic acids may be completely or partially located in the interior space of the liposome, within the bilayer membrane of the liposome, or associated with the exterior surface of the liposome membrane. The incorporation of a nucleic acid into liposomes is also referred to herein as “encapsulation” wherein the nucleic acid is entirely contained within the interior space of the liposome. The purpose of incorporating a mRNA into a transfer vehicle, such as a liposome, is often to protect the nucleic acid from an environment which may contain enzymes or chemicals that degrade nucleic acids and/or systems or receptors that cause the rapid excretion of the nucleic acids. Accordingly, in some embodiments, a suitable delivery vehicle is capable of enhancing the stability of the mRNA contained therein and/or facilitate the delivery of mRNA to the target cell or tissue.

Liposome Size

Suitable liposomes in accordance with the present invention may be made in various sizes. In some embodiments, provided liposomes may be made smaller than previously known mRNA encapsulating liposomes. In some embodiments, decreased size of liposomes is associated with more efficient delivery of mRNA. Selection of an appropriate liposome size may take into consideration the site of the target cell or tissue and to some extent the application for which the liposome is being made.

In some embodiments, an appropriate size of liposome is selected to facilitate systemic distribution of antibody encoded by the mRNA. In some embodiments, it may be desirable to limit transfection of the mRNA to certain cells or tissues. For example, to target hepatocytes a liposome may be sized such that its dimensions are smaller than the fenestrations of the endothelial layer lining hepatic sinusoids in the liver; in such cases the liposome could readily penetrate such endothelial fenestrations to reach the target hepatocytes.

Alternatively or additionally, a liposome may be sized such that the dimensions of the liposome are of a sufficient diameter to limit or expressly avoid distribution into certain cells or tissues. For example, a liposome may be sized such that its dimensions are larger than the fenestrations of the endothelial layer lining hepatic sinusoids to thereby limit distribution of the liposomes to hepatocytes.

In some embodiments, the size of a liposome is determined by the length of the largest diameter of the lipososme particle. In some embodiments, a suitable liposome has a size of or less than about 500 nm, 450 nm, 400 nm, 350 nm, 300 nm, 250 nm, 200 nm, 150 nm, 125 nm, 110 nm, 100 nm, 95 nm, 90 nm, 85 nm, 80 nm, 75 nm, 70 nm, 65 nm, 60 nm, 55 nm, or 50 nm. In some embodiments, a suitable liposome has a size no greater than about 250 nm (e.g., no greater than about 225 nm, 200 nm, 175 nm, 150 nm, 125 nm, 100 nm, 75 nm, or 50 nm). In some embodiments, a suitable liposome has a size ranging from about 10-250 nm (e.g., ranging from about 10-225 nm, 10-200 nm, 10-175 nm, 10-150 nm, 10-125 nm, 10-100 nm, 10-75 nm, or 10-50 nm). In some embodiments, a suitable liposome has a size ranging from about 100-250 nm (e.g., ranging from about 100-225 nm, 100-200 nm, 100-175 nm, 100-150 nm). In some embodiments, a suitable liposome has a size ranging from about 10-100 nm (e.g., ranging from about 10-90 nm, 10-80 nm, 10-70 nm, 10-60 nm, or 10-50 nm).

A variety of alternative methods known in the art are available for sizing of a population of liposomes. One such sizing method is described in U.S. Pat. No. 4,737,323, incorporated herein by reference. Sonicating a liposome suspension either by bath or probe sonication produces a progressive size reduction down to small ULV less than about 0.05 microns in diameter. Homogenization is another method that relies on shearing energy to fragment large liposomes into smaller ones. In a typical homogenization procedure, MLV are recirculated through a standard emulsion homogenizer until selected liposome sizes, typically between about 0.1 and 0.5 microns, are observed. The size of the liposomes may be determined by quasi-electric light scattering (QELS) as described in Bloomfield, Ann. Rev. Biophys. Bioeng., 10:421-150 (1981), incorporated herein by reference. Average liposome diameter may be reduced by sonication of formed liposomes. Intermittent sonication cycles may be alternated with QELS assessment to guide efficient liposome synthesis.

Pharmaceutical Compositions

To facilitate expression of mRNA in vivo, delivery vehicles such as liposomes can be formulated in combination with one or more additional nucleic acids, carriers, targeting ligands or stabilizing reagents, or in pharmacological compositions where it is mixed with suitable excipients. Techniques for formulation and administration of drugs may be found in “Remington's Pharmaceutical Sciences,” Mack Publishing Co., Easton, Pa., latest edition.

Provided liposomally-encapsulated or associated mRNAs, and compositions containing the same, may be administered and dosed in accordance with current medical practice, taking into account the clinical condition of the subject, the site and method of administration, the scheduling of administration, the subject's age, sex, body weight and other factors relevant to clinicians of ordinary skill in the art. The “effective amount” for the purposes herein may be determined by such relevant considerations as are known to those of ordinary skill in experimental clinical research, pharmacological, clinical and medical arts. In some embodiments, the amount administered is effective to achieve at least some stabilization, improvement or elimination of symptoms and other indicators as are selected as appropriate measures of disease progress, regression or improvement by those of skill in the art. For example, a suitable amount and dosing regimen is one that causes at least transient protein (e.g., enzyme) production.

Suitable routes of administration include, for example, oral, rectal, vaginal, transmucosal, pulmonary including intratracheal or inhaled, or intestinal administration; parenteral delivery, including intradermal, transdermal (topical), intramuscular, subcutaneous, intramedullary injections, as well as intrathecal, direct intraventricular, intravenous, intraperitoneal, and/or intranasal administration.

Alternately or additionally, liposomally encapsulated mRNAs and compositions of the invention may be administered in a local rather than systemic manner, for example, via injection of the pharmaceutical composition directly into a targeted tissue, preferably in a sustained release formulation. Local delivery can be affected in various ways, depending on the tissue to be targeted. For example, aerosols containing compositions of the present invention can be inhaled (for nasal, tracheal, or bronchial delivery); compositions of the present invention can be injected into the site of injury, disease manifestation, or pain, for example; compositions can be provided in lozenges for oral, tracheal, or esophageal application; can be supplied in liquid, tablet or capsule form for administration to the stomach or intestines, can be supplied in suppository form for rectal or vaginal application; or can even be delivered to the eye by use of creams, drops, or even injection. Formulations containing provided compositions complexed with therapeutic molecules or ligands can even be surgically administered, for example in association with a polymer or other structure or substance that can allow the compositions to diffuse from the site of implantation to surrounding cells. Alternatively, they can be applied surgically without the use of polymers or supports.

In some embodiments, provided liposomes and/or compositions are formulated such that they are suitable for extended-release of the mRNA contained therein. Such extended-release compositions may be conveniently administered to a subject at extended dosing intervals. For example, in one embodiment, the compositions of the present invention are administered to a subject twice day, daily or every other day. In a preferred embodiment, the compositions of the present invention are administered to a subject twice a week, once a week, every ten days, every two weeks, every three weeks, or more preferably every four weeks, once a month, every six weeks, every eight weeks, every other month, every three months, every four months, every six months, every eight months, every nine months or annually. Also contemplated are compositions and liposomes which are formulated for depot administration (e.g., intramuscularly, subcutaneously, intravitreally) to either deliver or release a mRNA over extended periods of time. Preferably, the extended-release means employed are combined with modifications made to the mRNA to enhance stability.

Also contemplated herein are lyophilized pharmaceutical compositions comprising one or more of the liposomes disclosed herein and related methods for the use of such compositions as disclosed for example, in U.S. Provisional Application No. 61/494,882, filed Jun. 8, 2011, the teachings of which are incorporated herein by reference in their entirety. For example, lyophilized pharmaceutical compositions according to the invention may be reconstituted prior to administration or can be reconstituted in vivo. For example, a lyophilized pharmaceutical composition can be formulated in an appropriate dosage form (e.g., an intradermal dosage form such as a disk, rod or membrane) and administered such that the dosage form is rehydrated over time in vivo by the individual's bodily fluids.

Provided liposomes and compositions may be administered to any desired tissue. In some embodiments, the mRNA delivered by provided liposomes or compositions is expressed in the tissue in which the liposomes and/or compositions were administered. In some embodiments, the mRNA delivered is expressed in a tissue different from the tissue in which the liposomes and/or compositions were administered Exemplary tissues in which delivered mRNA may be delivered and/or expressed include, but are not limited to the liver, kidney, heart, spleen, serum, brain, skeletal muscle, lymph nodes, skin, and/or cerebrospinal fluid.

According to various embodiments, the timing of expression of delivered mRNAs can be tuned to suit a particular medical need. In some embodiments, the expression of the protein encoded by delivered mRNA is detectable 1, 2, 3, 6, 12, 18, 24, 30, 36, 42, 48, 54, 60, 66, and/or 72 hours in serum or target tissues after a single administration of provided liposomes or compositions. In some embodiments, the expression of the protein encoded by the mRNA is detectable 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, and/or 7 days in serum or target tissues after a single administration of provided liposomes or compositions. In some embodiments, the expression of the protein encoded by the mRNA is detectable 1 week, 2 weeks, 3 weeks, and/or 4 weeks in serum or target tissues after a single administration of provided liposomes or compositions. In some embodiments, the expression of the protein encoded by the mRNA is detectable after a month or longer after a single administration of provided liposomes or compositions.

The present invention can be used to deliver mRNA at various doses. In some embodiments, an mRNA is administered at a dose ranging from about 0.1-5.0 mg/kg body weight, for example about 0.1-4.5, 0.1-4.0, 0.1-3.5, 0.1-3.0, 0.1-2.5, 0.1-2.0, 0.1-1.5, 0.1-1.0, 0.1-0.5, 0.1-0.3, 0.3-5.0, 0.3-4.5, 0.3-4.0, 0.3-3.5, 0.3-3.0, 0.3-2.5, 0.3-2.0, 0.3-1.5, 0.3-1.0, 0.3-0.5, 0.5-5.0, 0.5-4.5, 0.5-4.0, 0.5-3.5, 0.5-3.0, 0.5-2.5, 0.5-2.0, 0.5-1.5, or 0.5-1.0 mg/kg body weight. In some embodiments, an mRNA is administered at a dose of or less than about 5.0, 4.5, 4.0, 3.5, 3.0, 2.5, 2.0, 1.5, 1.0, 0.8, 0.6, 0.5, 0.4, 0.3, 0.2, or 0.1 mg/kg body weight.

While certain compounds, compositions and methods of the present invention have been described with specificity in accordance with certain embodiments, the following examples serve only to illustrate the compounds of the invention and are not intended to limit the same.

Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. The scope of the present invention is not intended to be limited to the above Description, but rather is as set forth in the following claims.