Human G protein-coupled receptor and modulators thereof for the treatment of cell death-related disorders

This application claims the benefit of priority from the following provisional applications, filed via U.S. Express mail with the United States Patent and Trademark Office on the indicated dates: U.S. Provisional No. 60/455,316, filed Mar. 17, 2003, and U.S. Provisional No. 60/532,001, filed Dec. 22, 2003. The disclosures of the foregoing applications are herein incorporated by reference in their entireties.

The present invention relates to methods of identifying whether a candidate compound is a modulator of a G protein-coupled receptor (GPCR). In certain embodiments, the GPCR is human. In certain embodiments, the GPCR is expressed endogenously by neuronal cells or muscle cells. In certain embodiments, the GPCR is neuroprotective or myoprotective. In certain embodiments, the GPCR is a Humanin receptor. The present invention also relates to methods of using a modulator of said GPCR. A preferred modulator is agonist. Agonists of the invention are useful as therapeutic agents for the prevention or treatment of neurodegenerative diseases in general, including Alzheimer's disease, Parkinson's disease, prion-associated disease, stroke and motor-neuron disease in particular, peripheral neuropathy, cerebral amyloid beta-protein angiopathy, and ischemic heart disease, including myocardial infarction and congestive heart failure.

A. Cell Death-Related Disorders

The following discussion is intended to facilitate the understanding of the invention, but is not intended nor admitted to be prior art to the invention.

Anomalous cell death is a component of a number of pathologies, including neurodegenerative disease, cerebral amyloid beta-protein angiopathy (CAA), and some cardiomyopathies. Said anomalous cell death may comprise apoptotic cell death. Agents that are cell death-protective have utility for the prevention or treatment of said pathologies. In particular, agents that are neuroprotective or myoprotective have utility for the prevention or treatment of said neurodegenerative disease or said myopathy.

Alzheimer's disease is a progressive neurodegenerative disease characterized by significant loss of function in more than one cognitive domain and often accompanied by changes in behavior or personality (Alzheimer's Disease; Decision Resources, Inc.; November 2001). Mutations in three different genes can cause early-onset familial Alzheimer's disease (FAD): amyloid precursor protein (APP), presenilin 1 (PS1) and presenilin 2 (PS2). Early-onset familial Alzheimer's disease accounts for less than 5% of all Alzheimer's disease cases, however. More typically, Alzheimer's disease occurs in people aged 65 and older. Alzheimer's disease is the most common cause of dementia, affecting approximately 20 million people worldwide. Amyloid beta-peptide (Aβ) is believed to play a central role in the pathology of Alzheimer's disease.

Mild cognitive impairment (MCI) is a state of cognitive impairment, most often of memory only, that is not severe enough to significantly impact occupational or social functioning. However, many people with MCI progress to Alzheimer's disease, at a rate of 10-15% per year and up to 60% within five years. Growing evidence shows that Alzheimer's disease pathology begins much earlier than the development of symptoms severe enough to allow a diagnosis of Alzheimer's criteria. In 2001, the American Academy of Neurology (AAN) published clinical practice parameters for the detection of MCI that will increase awareness, diagnosis, and treatment of both MCI and mild forms of Alzheimer's disease.

Parkinson's disease is a chronic, progressive neurodegenerative disorder affecting more than 2.7 million people in the United States, Japan, and Western Europe. Parkinson's disease is characterized by motor symptoms such as tremor, bradykinesia, muscle rigidity, gait dysfunction, and postural instability. Although researchers have identified degeneration of dopaminergic neurons in the substantia nigra as the primary pathophysiological mechanism, they believe that other neurotransmitter systems—such as the serontonergic and glutamatergic systems—are also intricately involved in the disease process.

Prion-associated diseases include, but are not held to be limited to, bovine spongiform encephalopathy, Creutzfeldt-Jakob disease, kuru, Gerstmann-Straussler-Scheinker syndrome, and fatal familial insomnia [Collins et al., Lancet (2004) 3:51-61].

Stroke is an acute interruption of blood flow in the brain caused by blood vessel obstruction or hemorrhage. Almost 90% of all strokes are ischemic; that is, they are due to a reduction in cerebral circulation below a critical threshold. Stroke accounts for 10-12% of medically related deaths in industrialized countries and is the third leading cause of death worldwide. Overall estimates of the annual number of strokes in the United States range from 500,000 to 700,000.

Motor neuron disease is characterized by a selective degeneration of the motor neurons of the spinal cord, brainstem, or motor cortex. Clinical subtypes are distinguished by the major site of degeneration. In amyotrophic lateral sclerosis there is involvement of upper, lower, and brainstem motor neurons. In progressive spinal muscular atrophy and related syndromes the motor neurons in the spinal cord are primarily affected. With progressive bulbar palsy, the initial degeneration occurs in the brainstem. In primary lateral sclerosis, the cortical neurons are affected in isolation.

Peripheral neuropathy is a broad term used to describe a variety of disorders which manifest as painful or uncomfortable sensations usually in the extremities and are of neuropathic origin. Nerve degeneration is believed to play a role in peripheral neuropathy. This condition is found commonly amongst end stage renal disease and diabetic patients. Distal symmetrical polyneuropathy (DSP)—a neuropathy affecting both sides of the body and attacking the distal sensory nerves—is the most common clinical manifestation of diabetic neuropathy and accounts for approximately 80-85% of all neuropathy observed in diabetic patients. DSP affects 30-40% of patients with type 1 and type 2 diabetes. Diabetic neuropathy can typically damage many different nerves, including the sciatic nerve.

Cerebral amyloid beta-protein (Aβ) angiopathy (CAA) is a key feature of Alzheimer's disease and related disorders. CAA is a disease of small blood vessels in the brain in which deposits of amyloid protein in the vessel walls may lead to stroke, brain hemorrhage, or dementia. Vascular amyloid deposition leads to degeneration of the vessel wall and aneurysm formation, and may be responsible for 10 to 15% of hemorrhagic strokes in the elderly.

Myocardial infarction is the damage or death of an area of heart muscle because of an inadequate supply of oxygen to that area. Myocardial infarctions are often caused by a clot that blocks one of the coronary arteries (the blood vessels that bring blood and oxygen to heart muscle). The clot prevents blood and oxygen from reaching that area of the heart, leading to the death of cardiomyocytes in that area.

Congestive heart failure affects nearly 5 million Americans with over 500,000 new cases diagnosed annually. Ischemic heart disease is the most common cause of congestive heart failure, accounting for 60-70% of all cases. By definition, congestive heart failure is a clinical syndrome in which heart disease reduces cardiac output, increases venous pressures, and is accompanied by molecular abnormalities that cause progressive deterioration of the failing heart and premature cardiomyocyte (cardiac muscle cell) death. Any definition of heart failure that does not consider the molecular processes that accelerate myocardial death overlooks a major clinical feature of this syndrome. Evidence in mice and rats demonstrate that inhibitors of cardiomyocyte death pathways significantly improve cardiac function and animal survival [Laugwitz et al., Hum Gene Ther (2001) 12:2051-63].

B. Humanin

The following discussion is intended to facilitate the understanding of the invention, but is not intended nor admitted to be prior art to the invention.

Humanin polypeptide was originally identified through its capacity to protect neuronal cells from Alzheimer's disease-relevant toxicity [Hashimoto et al., Biochem Biophys Res Commun (2001) 283:460-8]. Humanin has the amino acid sequence: Met-Ala-Pro-Arg-Gly-Phe-Ser-Cys-Leu-Leu-Leu-Leu-Thr-Ser-Glu-Ile-Asp-Leu-Pro-Val-Lys-Arg-Arg-Ala (SEQ ID NO. 15) (GenBank® Accession No. AAK50430). An analog of Humanin having a substitution of glycine for serine at amino acid position 14, [Gly14]-Humanin, has been reported to be significantly more active than Humanin [Niikura et al., J Neurosci Res (2002) 70:380-91]. Humanin immunoreactivity has been detected in an Alzheimer's disease brain, but little in normal human brains [Tajima et al., Neurosci Lett (2002) 324:227-31].

Humanin has been found to protect neuronal cells from a number of toxic insults. This includes neurotoxicity mediated by three mutant genes that cause FAD as well as by Aβ [Hashimoto et al., Proc Natl Acad Sci USA (2001) 98:6336-41; Niikura et al., J Neurosci Res (2002) 70:380-91]. Aβ neurotoxicity involves a c-Jun N-terminal kinase (JNK) pathway, and Humanin acts at a point downstream of JNK [Hashimoto et al., J Neurochem (2003) 84:864-77]. Humanin has also been reported to have protective activity for neurons against serum deprivation [Takahashi et al., Neuroreport (2002) 13:903-7]. The rat ortholog of Humanin has been reported to have protective activity for neurons against excitotoxic death [Caricasole et al., FASEB J (2002) 16:1331-3].

Humanin has also been shown to rescue cortical neurons from prion-peptide-induced apoptosis [Sponne et al., Mol Cell Neurosci (2004) 25:95-102].

Humanin has further been shown to improve learning and memory impairment in mice, thereby evidencing utility as a beneficial agent for the prevention or treatment of said learning or memory impairment [Mamiya & Ukai, Br J Pharmacol (2001) 134:1597-9].

Recently, Humanin has been shown to be protective for muscle cells. Humanin rescues human cerebrovascular smooth muscle cells from Aβ-induced toxicity.

Collectively, the foregoing indicates that Humanin and compounds evidencing Humanin activity are cell death-protective. Said cell death may comprise apoptotic cell death. In particular, Humanin and compounds evidencing Humanin activity protect neuronal cells and muscle cells from cell death and have utility for the prevention or treatment of neurodegenerative disease, cerebral amyloid angiopathy, and some cardiomyopathies.

C. G Protein-Coupled Receptors

The following discussion is intended to facilitate the understanding of the invention, but is not intended nor admitted to be prior art to the invention.

Although a number of receptor classes exist in humans, by far the most abundant and therapeutically relevant is represented by the G protein-coupled receptor (GPCR) class. It is estimated that there are some 30,000-40,000 genes within the human genome, and of these, approximately 2% are estimated to code for GPCRs.

GPCRs represent an important area for the development of pharmaceutical products: from approximately 20 of the 100 known GPCRs, approximately 60% of all prescription pharmaceuticals have been developed. For example, in 1999, of the top 100 brand name prescription drugs, the following drugs interact with GPCRs (the primary diseases and/or disorders treated related to the drug is indicated in parentheses):

GPCRs share a common structural motif, having seven sequences of between 22 to 24 hydrophobic amino acids that form seven alpha helices, each of which spans the membrane (each span is identified by number, i.e., transmembrane-1 (TM-1), transmembrane-2 (TM-2), etc.). The transmembrane helices are joined by strands of amino acids between transmembrane-2 and transmembrane-3, transmembrane-4 and transmembrane-5, and transmembrane-6 and transmembrane-7 on the exterior, or “extracellular” side, of the cell membrane (these are referred to as “extracellular” regions 1, 2 and 3 (EC-1, EC-2 and EC-3), respectively). The transmembrane helices are also joined by strands of amino acids between transmembrane-1 and transmembrane-2, transmembrane-3 and transmembrane-4, and transmembrane-5 and transmembrane-6 on the interior, or “intracellular” side, of the cell membrane (these are referred to as “intracellular” regions 1, 2 and 3 (IC-1, IC-2 and IC-3), respectively). The “carboxy” (“C”) terminus of the receptor lies in the intracellular space within the cell, and the “amino” (“N”) terminus of the receptor lies in the extracellular space outside of the cell.

Generally, when a ligand binds with the receptor (often referred to as “activation” of the receptor), there is a change in the conformation of the receptor that facilitates coupling between the intracellular region and an intracellular “G-protein.” It has been reported that GPCRs are “promiscuous” with respect to G proteins, i.e., that a GPCR can interact with more than one G protein. See, Kenakin, T., 43 Life Sciences 1095 (1988). Although other G proteins exist, currently, Gq, Gs, Gi, Gz and Go are G proteins that have been identified. Ligand-activated GPCR coupling with the G-protein initiates a signaling cascade process (referred to as “signal transduction”). Under normal conditions, signal transduction ultimately results in cellular activation or cellular inhibition. Although not wishing to be bound to theory, it is thought that the IC-3 loop as well as the carboxy terminus of the receptor interact with the G protein.

There are also promiscuous G proteins, which appear to couple several classes of GPCRs to the phospholipase C pathway, such as Gα15 or Gα16 [Offermanns & Simon, J Biol Chem (1995) 270:15175-80], or chimeric G proteins designed to couple a large number of different GPCRs to the same pathway, e.g. phospholipase C [Milligan & Rees, Trends in Pharmaceutical Sciences (1999) 20:118-24].

Gi-coupled GPCRs lower intracellular cAMP levels. A Gi-like G protein is a G protein that similarly leads to lower intracellular cAMP level. By this criterion, Go and Gz are Gi-like G proteins. The melanophore technology (see infra) is useful for identifying Gi-coupled GPCRs and also for identifying modulators of said Gi-coupled GPCRs.

Under physiological conditions, GPCRs exist in the cell membrane in equilibrium between two different conformations: an “inactive” state and an “active” state. A receptor in an inactive state is unable to link to the intracellular signaling transduction pathway to initiate signal transduction leading to a biological response. Changing the receptor conformation to the active state allows linkage to the transduction pathway (via the G-protein) and produces a biological response.

A receptor may be stabilized in an active state by a ligand or a compound such as a drug. Recent discoveries, including but not exclusively limited to modifications to the amino acid sequence of the receptor, provide means other than ligands or drugs to promote and stabilize the receptor in the active state conformation. These means effectively stabilize the receptor in an active state by simulating the effect of a ligand binding to the receptor. Stabilization by such ligand-independent means is termed “constitutive receptor activation.”

D. FPRL2

The following discussion is intended to facilitate the understanding of the invention, but is not intended nor admitted to be prior art to the invention.

FPRL2 (GenBank® Accession No. L14061) is a GPCR for which no endogenous ligand has yet been demonstrated. The synthetic peptide WKYMVm (SEQ ID NO: 16) recently has been shown to bind to FPRL2 on mature human dendritic cells and elicit an increase in intracellular Ca 2+ [Yang et al., J Leukoc Biol (2002) 72:598-607].

FPRL2 is most similar to the GPCRs FPRL1 (GenBank® Accession No. AF054013) and FPR (GenBank® Accession No. M60627). Recently, it was reported that Humanin uses FPRL1 as a functional receptor [Ying et al., Journal of Interferon and Cytokine Research (2002) 22 (Supplement 1):S180]. No endogenous ligand for FPRL1 or FPR has been shown to also be a ligand for FPRL2 and, in fact, a number of endogenous ligands for FPRL1 or FPR (lipoxin A4, e.g.) have been shown not to be ligands for FPRL2.

Applicants have unexpectedly identified Humanin as a selective agonist of FPRL2 and thus provide Humanin as a first endogenous ligand of FPRL2. Applicants have further unexpectedly identified FPRL2 as a receptor selectively engaged by Humanin.

The present invention is directed in part to methods of identifying whether a test compound is a modulator of a Humanin GPCR, wherein said Humanin GPCR is FPRL2. The invention also encompasses modulators identified by said methods as well as methods of using said modulators for the prevention or treatment of cell death-related disorders, in particular disorders related to the death of neuronal cells or muscle cells.

In a first aspect, the invention features a method of identifying whether a candidate compound is a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein the receptor couples to a G protein; comprising the steps of:

(a) contacting the candidate compound with the receptor; (b) determining whether the receptor functionality is modulated;

wherein a change in receptor functionality is indicative of the candidate compound being a modulator of a Humanin GPCR.

In certain embodiments, said Humanin GPCR is recombinant. In certain embodiments, said contacting comprises contacting with a host cell or with membrane of a host cell that expresses the GPCR, wherein said host cell comprises an expression vector comprising a polynucleotide encoding the receptor.

In some embodiments, said contacting is carried out in the presence of a known ligand of the GPCR. In some embodiments, said known ligand is an agonist of the GPCR. In some embodiments, said agonist is Humanin or a peptide analog or derivative thereof. In some embodiments, said analog is [Gly14]-Humanin. In certain embodiments, said presence of said Humanin or said [Gly14]-Humanin is at about EC50 to about EC75 for the means of said determining. By way of example and not limitation, EC50 is about 0.32 μM and EC75 is about 1.5 μM for Humanin in [ 35 S]GTPγS binding assay. Humanin and [Gly14]-Humanin are commercially available (Peptides International, Louisville, Ky.). In some embodiments, said agonist is Compound 1 (“Cmpd 1” in Table 1). In certain embodiments, said presence of said Compound 1 is at about EC50 to about EC75 for the means of said determining. By way of example and not limitation, EC50 is about 2.8 μM and EC75 is about 15 μM for Compound 1 in [ 35 S]GTPγS binding assay.

The invention also relates to a method of identifying whether a candidate compound is a modulator of a cell death-protective GPCR, comprising the steps of:

(a) contacting the candidate compound with a GPCR comprising an FPRL2 amino acid sequence, wherein the receptor couples to a G protein; and (b) determining whether the receptor functionality is modulated;

wherein a change in receptor functionality is indicative of the candidate compound being a modulator of a cell death-protective GPCR.

In certain embodiments, said GPCR is recombinant. In certain embodiments, said contacting comprises contacting with a host cell or with membrane of a host cell that expresses the GPCR, wherein said host cell comprises an expression vector comprising a polynucleotide encoding the receptor.

In certain embodiments, said cell is neuronal and said GPCR is neuroprotective. In certain embodiments, said cell is muscle and said GPCR is myoprotective. In some embodiments, said cell is non-neuronal and non-muscle. In certain embodiments, said cell expresses human FPRL2.

In some embodiments, said cell death comprises apoptotic cell death. In some embodiments, said cell death comprises cell death that is Aβ-induced. In some embodiments, said cell death comprises cell death that is JNK activation-dependent. In some embodiments, said cell death comprises cell death induced by deprivation of a serum component. In some embodiments, said cell death comprises excitotoxic cell death.

In some embodiments, said contacting is carried out in the presence of a known ligand of the GPCR. In some embodiments, said known ligand is an agonist of the GPCR. In some embodiments, said agonist is Humanin or a peptide analog or derivative thereof. In some embodiments, said analog is [Gly14]-Humanin. In certain embodiments, said presence of said Humanin or said [Gly14]-Humanin is at about EC50 to about EC75 for the means of said determining. By way of example and not limitation, EC50 is about 0.32 μM and EC75 is about 1.5 μM for Humanin in [ 35 S]GTPγS binding assay. Humanin and [Gly14]-Humanin are commercially available (Peptides International, Louisville, Ky.). In some embodiments, said agonist is Compound 1. In certain embodiments, said presence of said Compound 1 is at about EC50 to about EC75 for the means of said determining. By way of example and not limitation, EC50 is about 2.8 μM and EC75 is about 15 μM for Compound 1 in [ 35 S]GTPγS binding assay.

The invention also relates to a method of identifying whether a candidate compound is a modulator of neuroprotection, comprising the steps of:

(a) contacting the candidate compound with a GPCR comprising an FPRL2 amino acid sequence, wherein the receptor couples to a G protein; and (b) determining whether the receptor functionality is modulated;

wherein a change in receptor functionality is indicative of the candidate compound being a modulator of neuroprotection.

In certain embodiments, said GPCR is recombinant. In certain embodiments, said contacting comprises contacting with a host cell or with membrane of a host cell that expresses the GPCR, wherein said host cell comprises an expression vector comprising a polynucleotide encoding the receptor.

In certain embodiments, an increase in receptor functionality is indicative of the candidate compound being a neuroprotective compound.

In some embodiments, said contacting is carried out in the presence of a known ligand of the GPCR. In some embodiments, said known ligand is an agonist of the GPCR. In some embodiments, said agonist is Humanin or a peptide analog or derivative thereof. In some embodiments, said analog is [Gly14]-Humanin. In certain embodiments, said presence of said Humanin or said [Gly14]-Humanin is at about EC50 to about EC75 for the means of said determining. By way of example and not limitation, EC50 is about 0.32 μM and EC75 is about 1.5 μM for Humanin in [ 35 S]GTPγS binding assay. Humanin and [Gly14]-Humanin are commercially available (Peptides International, Louisville, Ky.). In some embodiments, said agonist is Compound 1. In certain embodiments, said presence of said Compound 1 is at about EC50 to about EC75 for the means of said determining. By way of example and not limitation, EC50 is about 2.8 μM and EC75 is about 15 μM for Compound 1 in [ 35 S]GTPγS binding assay.

The invention also relates to a method of identifying whether a candidate compound is a modulator of myoprotection, comprising the steps of:

(a) contacting the candidate compound with a GPCR comprising an FPRL2 amino acid sequence, wherein the receptor couples to a G protein; and (b) determining whether the receptor functionality is modulated;

wherein a change in receptor functionality is indicative of the candidate compound being a modulator of myoprotection.

In certain embodiments, said GPCR is recombinant. In certain embodiments, said contacting comprises contacting with a host cell or with membrane of a host cell that expresses the GPCR, wherein said host cell comprises an expression vector comprising a polynucleotide encoding the receptor.

In certain embodiments, an increase in receptor functionality is indicative of the candidate compound being a myoprotective compound.

In some embodiments, said contacting is carried out in the presence of a known ligand of the GPCR. In some embodiments, said known ligand is an agonist of the GPCR. In some embodiments, said agonist is Humanin or a peptide analog or derivative thereof. In some embodiments, said analog is [Gly14]-Humanin. In certain embodiments, said presence of said Humanin or said [Gly14]-Humanin is at about EC50 to about EC75 for the means of said determining. By way of example and not limitation, EC50 is about 0.32 μM and EC75 is about 1.5 μM for Humanin in [ 35 S]GTPγS binding assay. Humanin and [Gly14]-Humanin are commercially available (Peptides International, Louisville, Ky.). In some embodiments, said agonist is Compound 1. In certain embodiments, said presence of said Compound 1 is at about EC50 to about EC75 for the means of said determining. By way of example and not limitation, EC50 is about 2.8 μM and EC75 is about 15 μM for Compound 1 in [ 35 S]GTPγS binding assay.

The invention also relates to a method of identifying whether a candidate compound is a modulator of a Humanin GPCR, comprising the steps of:

(a) culturing Humanin GPCR-expressing host cells under conditions that would allow expression of a recombinant Humanin GPCR, said host cells being transfected with an expression vector comprising a polynucleotide encoding said recombinant Humanin GPCR comprising an FPRL2 amino acid sequence, wherein the receptor couples to a G protein; (b) contacting the Humanin GPCR-expressing host cells of step (a) or membrane of said host cells of step (a) with the candidate compound; (c) contacting control host cells or membrane of said control host cells with the candidate compound of step (b), wherein said control host cells do not express recombinant Humanin GPCR protein; (d) measuring the modulating effect of the candidate compound which interacts with the recombinant Humanin GPCR from the host cells of step (a) and control host cells of step (c); and (e) comparing the modulating effect of the test compound on the host cells and control host cells.

In certain embodiments, said transfected host cells are transiently transfected. In certain embodiments, said host cells are stably transfected.

The invention also relates to a method of identifying whether a candidate compound is a modulator of a Humanin GPCR, comprising the steps of:

(a) culturing Humanin GPCR-expressing host cells under conditions that would allow expression of a recombinant Humanin GPCR, said host cells being transfected with an expression vector comprising a polynucleotide encoding said recombinant Humanin GPCR comprising an FPRL2 amino acid sequence, wherein the receptor couples to a G protein; (b) contacting a first population of Humanin GPCR-expressing cells of step (a) or membrane of said first population of Humanin GPCR-expressing cells of step (a) with a known ligand of said Humanin GPCR; (c) contacting a second population of Humanin GPCR-expressing cells of step (a) or membrane of said second population of Humanin GPCR-expressing cells of step (a) with the candidate compound and with the known Humanin GPCR ligand; (d) contacting control host cells or membrane of said control host cells with the candidate compound of step (c), wherein said control host cells do not express recombinant Humanin GPCR protein; (e) measuring the modulating effect of the candidate compound, which interacts with recombinant Humanin GPCR, in the presence and absence of the known Humanin GPCR ligand, from the cells of step (b), step (c) and step (d); and (f) comparing the modulating effect of the candidate compound as determined from step (b), step (c) and step (d).

In some embodiments, said ligand is an agonist of said Humanin GPCR. In some embodiments, said agonist is Humanin or a peptide analog or derivative thereof. In some embodiments, said analog is [Gly14]-Humanin. In certain embodiments, said presence of said Humanin or said [Gly14]-Humanin is at about 0.1 μM to 1.5 μM, more preferably at about 0.5 μM to 1 μM. In certain embodiments, said presence of said agonist is at EC50-EC75 for the means of said determining. By way of example and not limitation, EC50 is about 0.3 μM and EC75 is about 1.5 μM for Humanin in [ 35 S]GTPγS binding assay. In some embodiments, said agonist is Compound 1. In certain embodiments, said presence of said Compound 1 is at about EC50 to about EC75 for the means of said determining. By way of example and not limitation, EC50 is about 2.8 μM and EC75 is about 15 μM for Compound 1 in [ 35 S]GTPγS binding assay.

In certain embodiments, said transfected host cells are transiently transfected. In certain embodiments, said host cells are stably transfected.

The invention also relates to a method of identifying whether a candidate compound is a modulator of a Humanin GPCR, comprising the steps of:

(a) culturing Humanin GPCR-expressing host cells under conditions that would allow expression of a recombinant Humanin GPCR, said host cells being transfected with an expression vector comprising a polynucleotide encoding said recombinant Humanin GPCR comprising an FPRL2 amino acid sequence, wherein the receptor couples to a G protein; (b) contacting a first population of the Humanin GPCR-expressing host cells of step (a) or membrane of said first population of the Humanin GPCR-expressing host cells of step (a) with the candidate compound; (c) not contacting a second population of the Humanin GPCR-expressing cells of step (a) or membrane of said second population of the Humanin GPCR-expressing cells of step (a) with the candidate compound of step (b); (d) contacting control host cells or membrane of said control host cells to the candidate compound of step (b), wherein said control host cells do not express recombinant Humanin GPCR protein; (e) measuring the modulating effect of the candidate compound, which interacts with recombinant Humanin GPCR protein, from the cells of step (b) and step (c) and from the cells of step (d); and (f) comparing the modulating effect of the candidate compound as determined from step (b) and step (c) and from step (d).

In certain embodiments, said transfected host cells are transiently transfected. In certain embodiments, said transfected host cells are stably transfected.

In some embodiments, the FPRL2 amino acid sequence is the amino acid sequence of SEQ ID NO:2. In some embodiments, the FPRL2 amino acid sequence is a variant of the amino acid sequence of SEQ ID NO:2. In some embodiments, said variant of the amino acid sequence of SEQ ID NO:2 is an allelic variant or mammalian ortholog of said amino acid sequence. In some embodiments, said variant of the amino acid sequence of SEQ ID NO:2 is a non-endogenous, constitutively activated mutant of said amino acid sequence or of an allelic variant or mammalian ortholog of said amino acid sequence. In certain embodiments, said non-endogenous, constitutively activated mutant is the amino acid sequence of SEQ ID NO:12. In certain embodiments, said variant of the amino acid sequence of SEQ ID NO:2 is a biologically active fragment of said amino acid sequence or of an allelic variant or mammalian ortholog of said amino acid sequence. In certain embodiments, said biologically active fragment is amino acids 2-353 of the amino acid sequence of SEQ ID NO:2 or of an allelic variant or mammalian ortholog of said amino acid sequence. In certain embodiments, said variant of the amino acid sequence of SEQ ID NO:2 is at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% identical to the amino acid sequence of SEQ ID NO:2. In some embodiments, said variant of the amino acid sequence of SEQ ID NO:2 is at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% identical to the amino acid sequence of SEQ ID NO:2.

In certain embodiments, said GPCR comprising an FPRL2 amino acid sequence is a fusion protein comprising one or more epitope tags. In certain embodiments, said fusion protein comprising one or more epitope tags is the amino acid sequence of SEQ ID NO:6.

In certain embodiments, said G protein is pertussis toxin sensitive. In certain embodiments, said G protein is Gi or Go. In certain embodiments, said G protein is Gi. In certain embodiments, said G protein is Go.

In certain embodiments, said G protein is Gα15 or Gα16. In certain embodiments, said G protein is Gα15. In certain embodiments, said G protein is Gα16.

In certain embodiments, said G protein is Gq.

In other certain embodiments, said determining or said comparing is through the use of a Melanophore assay.

In certain embodiments, said determining or said comparing is through the measurement of the level of a second messenger selected from the group consisting of cyclic AMP (cAMP), cyclic GMP (cGMP), inositol triphosphate (IP 3 ), diacylglycerol (DAG), and Ca 2+ . In further preferred embodiments, said second messenger is cAMP. In certain embodiments, the level of the cAMP is reduced. In some embodiments, said measurement of cAMP is carried out with membrane comprising said GPCR.

In certain embodiments, said determining or said comparing is through CRE-reporter assay. In certain embodiments, said reporter is luciferase. In some embodiments, said reporter is β-galactosidase.

In certain embodiments, said determining or said comparing is through measurement of intracellular Ca 2+ . In certain embodiments, the level of intracellular Ca 2+ is increased. In certain embodiments, said Ca 2+ measurement is carried out by FLIPR.

In certain embodiments, said host cell comprises Gα15 or Gα16 or chimeric Gq(del)/Gi alpha subunit and said determining or said comparing is through measurement of intracellular Ca 2+ . In certain embodiments, said host cell comprises Gα15 and said determining or said comparing is through measurement of intracellular Ca 2+ . In certain embodiments, said host cell comprises Gα16 and said determining or said comparing is through measurement of intracellular Ca 2+ . In certain embodiments, said host cell comprises chimeric Gq(del)/Gi alpha subunit and said determining or said comparing is through measurement of intracellular Ca 2+ .

In certain embodiments, said determining or said comparing is through measurement of intracellular IP 3 . In certain embodiments, the level of intracellular IP 3 is increased.

In certain embodiments, said host cell comprises Gα15 or Gα16 or chimeric Gq(del)/Gi alpha subunit and said determining or said comparing is through measurement of intracellular IP 3 . In certain embodiments, said host cell comprises Gα15 and said determining or said comparing is through measurement of intracellular IP 3 . In certain embodiments, said host cell comprises Gα16 and said determining or said comparing is through measurement of intracellular IP 3 . In certain embodiments, said host cell comprises chimeric Gq(del)/Gi alpha subunit and said determining or said comparing is through measurement of intracellular IP 3 .

In certain embodiments, said determining or said comparing is through the measurement of GTPγS binding to membrane comprising said GPCR. In certain embodiments, said GTPγS is labeled with [ 35 S].

In certain embodiments, said method further comprises the step of comparing the modulation of the receptor caused by the candidate compound to a second modulation of the receptor caused by contacting the receptor with a known modulator of the receptor. In certain embodiments, said known modulator is an agonist. In certain embodiments, said agonist is Humanin or a peptide analog or derivative thereof. In certain embodiments, said analog of Humanin is [Gly14]-Humanin. In certain embodiments, said agonist is Compound 1.

In a second aspect, the invention features a compound, wherein said compound is a Dimethyl octahydro-phenanthrene carboxylic acid amide derivative of Formula (I):

wherein:

R 1 -R 4 are each independently selected form the group consisting of H, C 1-5 acyl, C 1-5 acyloxy, C 2-6 alkenyl, C 1-4 alkoxy, C 1-6 alkyl, C 1-4 alkylcarboxamide, C 2-6 alkynyl, C 1-4 alkylamino, C 1-4 alkylsulfonamide, C 1-4 alkylsulfinyl, C 1-4 alkylsulfonyl, C 1-4 alkylthio, C 1-4 alkylureyl, amino, carbo-C 1-6 -alkoxy, carboxamide, carboxy, cyano, C 3-6 cycloalkyl, C 2-6 dialkylamino, C 2-6 dialkylcarboxamide, halogen, C 1-4 haloalkoxy, C 1-4 haloalkyl, C 1-4 haloalkylsulfinyl, C 1-4 haloalkylsulfonyl, C 1-4 haloalkylthio, hydroxyl, nitro and thiol; or a pharmaceutically acceptable salt, solvate or hydrate thereof.

In certain embodiments, at least one R 1 -R 4 group is not H.

In certain embodiments, when R 1 , R 3 , and R 4 are each H, then R 2 is not —OAc or —OCH 3 .

In certain embodiments, when R 1 , and R 4 are both H, and R 3 is —CH 3 , then R 2 is not —OH.

In certain embodiments, when R 1 , and R 4 are both H, and R 2 is H or Br, then R 3 is not —CH(CH 3 ) 2 .

In certain embodiments, when R 1 , R 2 , and R 3 are each H, then R is not —OAc.

In certain embodiments, R 1 is H or halogen. In certain embodiments, R 1 is H and can be represented by Formula (Ib) as shown below:

wherein each variable in Formula (Ib) has the same meaning as described herein, supra and infra.

In certain embodiments, R 4 is H or halogen. In certain embodiments, R 4 is H and can be represented by Formula (Id) as shown below:

wherein each variable in Formula (Id) has the same meaning as described herein, supra and infra. In certain embodiment, compound of the present invention are of Formula (Id) wherein R 1 is H; these compound can be represented by Formula (If) as shown below:

wherein each variable in Formula (Id) has the same meaning as described herein, supra and infra.

In certain embodiment, R 2 is selected form the group consisting of H, C 1-5 acyl, C 1-5 acyloxy, C 2-6 alkenyl, C 1-4 alkoxy, C 1-6 alkyl, C 1-4 alkylcarboxamide, C 1-4 alkylsulfonamide, C 1-4 alkylsulfinyl, C 1-4 alkylsulfonyl, C 1-4 alkylthio, C 2-6 dialkylamino, halogen, hydroxyl, nitro, and thiol.

In certain embodiment, R 2 is selected form the group consisting of H, C 2-6 alkynyl, C 1-4 alkylamino, C 1-4 alkylureyl, amino, carbo-C 1-6 -alkoxy, carboxamide, carboxy, cyano, C 3-6 cycloalkyl, C 2-6 dialkylcarboxamide, C 1-4 haloalkoxy, C 1-4 haloalkyl, C 1-4 haloalkylsulfinyl, C 1-4 haloalkylsulfonyl, and C 1-4 haloalkylthio.

In certain embodiment, R 2 is H or halogen. In some embodiments, R 2 is H.

In certain embodiment, R 3 is selected form the group consisting of H, C 1-5 acyl, C 1-5 acyloxy, C 2-6 alkenyl, C 1-4 alkoxy, C 1-6 alkyl, C 1-4 alkylcarboxamide, C 1-4 alkylsulfonamide, C 1-4 alkylsulfinyl, C 1-4 alkylsulfonyl, C 1-4 alkylthio, C 2-6 dialkylamino, halogen, hydroxyl, nitro, and thiol. In certain embodiment, R 3 is C 1-6 alkyl. In certain embodiment, R 3 is selected form the group consisting of H, C 2-6 alkynyl, C 1-4 alkylamino, C 1-4 alkylureyl, amino, carbo-C 1-6 -alkoxy, carboxamide, carboxy, cyano, C 3-6 cycloalkyl, C 2-6 dialkylcarboxamide, C 1-4 haloalkoxy, C 1-4 haloalkyl, C 1-4 haloalkylsulfinyl, C 1-4 haloalkylsulfonyl, and C 1-4 haloalkylthio.

In a third aspect, the invention features a compound of Formula (I):

wherein:

R 1 -R 4 are each independently selected form the group consisting of H, C 1-5 acyl, C 1-5 acyloxy, C 2-6 alkenyl, C 1-4 alkoxy, C 1-6 alkyl, C 1-4 alkylcarboxamide, C 2-6 alynyl, C 1-4 alkylamino, C 1-4 alkylsulfonamide, C 1-4 alkylsulfinyl, C 1-4 alkylsulfonyl, C 1-4 alkylthio, C 1-4 alkylureyl, amino, carbo-C 1-6 -alkoxy, carboxamide, carboxy, cyano, C 3-6 cycloalkyl, C 1-6 dialkylamino, C dialkylcarboxamide, halogen, C 1-4 haloalkoxy, C 1-4 haloalkyl, C 1-4 haloalkylsulfinyl, C 1-4 haloalkylsulfonyl, C 1-4 haloalkylthio, hydroxyl, nitro and thiol; or a pharmaceutically acceptable salt, solvate or hydrate thereof.

In a fourth aspect, the invention features a modulator of a GPCR identified according to a method of the first aspect, provided that the modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. The invention also features a modulator of a GPCR identified according to a method of the first aspect, provided that the modulator is not a peptide.

The invention also features a modulator of a GPCR identifiable according to a method of the first aspect, provided that the modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. The invention also features a modulator of a GPCR identifiable according to a method of the first aspect, provided that the modulator is not a peptide.

In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist and antagonist. In certain embodiments, said modulator is an agonist. In certain embodiments, said modulator is a partial agonist. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist.

In certain embodiments, said modulator is preferably an agonist. In certain embodiments, said agonist is a compound according to the second aspect. In certain embodiments, said agonist is a compound according to the third aspect.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 mM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

In some embodiments, said modulator is selective for the GPCR.

In some embodiments, said modulator modulates a Humanin GPCR.

In some embodiments, said modulator is cell death-protective.

In some embodiments, said modulator is neuroprotective.

In some embodiments, said modulator is myoprotective.

In some embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In some embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In a fifth aspect, the invention features a method of preparing a pharmaceutical or physiologically acceptable composition comprising admixing a carrier and a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence. In certain embodiments, the modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, the modulator is not a peptide. In certain embodiments, the modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain embodiments, the modulator is an agonist. In certain embodiments, the modulator is a partial agonist. In certain embodiments, the modulator is an inverse agonist. In certain embodiments, the modulator is an antagonist. In certain embodiments, the modulator is preferably an agonist. In certain embodiments, said agonist is a compound according to the second aspect. In certain embodiments, said agonist is a compound according to the third aspect.

The invention also features a method of preparing a pharmaceutical or physiologically acceptable composition which comprises identifying a modulator of a Humanin GPCR, wherein said receptor comprises an FPRL2 amino acid sequence, and then admixing a carrier and the modulator, wherein the modulator is identifiable by a method according to a method of the first aspect. In certain embodiments, the modulator is identified according to a method of the first aspect. In certain embodiments, the modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, the modulator is not a peptide. In certain embodiments, the modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain embodiments, the modulator is an agonist. In certain embodiments, the modulator is a partial agonist. In certain embodiments, the modulator is an inverse agonist. In certain embodiments, the modulator is an antagonist. In certain embodiments, the modulator is preferably an agonist. In certain embodiments, said agonist is a compound according to the second aspect. In certain embodiments, said agonist is a compound according to the third aspect.

In certain embodiments, said composition is pharmaceutical. In certain embodiments, said composition is physiologically acceptable.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC 50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

In some embodiments, said modulator is selective for the GPCR.

In some embodiments, said modulator modulates a Humanin GPCR.

In some embodiments, said modulator is cell death-protective.

In some embodiments, said modulator is neuroprotective.

In some embodiments, said modulator is myoprotective.

In some embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In some embodiments, said orally bioavailable modulator is finder able to cross the blood-brain barrier.

In a sixth aspect, the invention features a method of modulating the activity of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, comprising the step of contacting the receptor with a modulator of the receptor. In certain embodiments, the modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, the modulator is not a peptide. In certain embodiments, the modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain embodiments, the modulator is an agonist. In certain embodiments, the modulator is a partial agonist. In certain embodiments, the modulator is an inverse agonist. In certain embodiments, the modulator is an antagonist. In certain embodiments, the modulator is preferably an agonist. In certain embodiments, said agonist is a compound according to the second aspect. In certain embodiments, said agonist is a compound according to the third aspect.

The invention also features a method of modulating the activity of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, comprising the step of contacting the receptor with a modulator of the receptor, wherein the modulator is identifiable by a method of the first aspect. In certain embodiments, the modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, the modulator is not a peptide. In certain embodiments, the modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain embodiments, the modulator is an agonist. In certain embodiments, the modulator is a partial agonist. In certain embodiments, the modulator is an inverse agonist. In certain embodiments, the modulator is an antagonist. In certain embodiments, the modulator is preferably an agonist. In certain embodiments, said agonist is a compound according to the second aspect. In certain embodiments, said agonist is a compound according to the third aspect.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

In some embodiments, said modulator is selective for the GPCR.

In some embodiments, said modulator modulates a Humanin GPCR.

In some embodiments, said modulator is cell death-protective.

In some embodiments, said modulator is neuroprotective.

In some embodiments, said modulator is myoprotective.

In some embodiments, said modulator opposes protection from cell death.

In some embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In some embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said contacting comprises administration of the modulator to a membrane comprising the receptor.

In certain embodiments, said contacting comprises administration of the modulator to a cell comprising the receptor.

In certain embodiments, said contacting comprises administration of the modulator to a tissue comprising the receptor.

In certain embodiments, said contacting comprises administration of the modulator to an individual comprising the receptor. In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In certain embodiments, said administration to an individual is oral.

In some embodiments, said modulator is an agonist and said individual is in need of prevention of or treatment for cell death, wherein said cell is non-neuronal and non-muscle.

In some embodiments, said modulator is an agonist and said individual is in need of prevention of or treatment for a cell death-related disorder, wherein said cell is non-neuronal and non-muscle.

In some embodiments, said modulator is an inverse agonist or an antagonist and said individual is in need of opposition to protection from cell death, wherein said cell is non-neuronal and non-muscle.

In some embodiments, said modulator is an inverse agonist or an antagonist and said individual is in need of prevention of or treatment for a cell proliferative disorder, wherein said cell is non-neuronal and non-muscle.

In certain embodiments, said modulator is an agonist and said individual is in need of reduction of neuronal cell death.

In certain embodiments, said modulator is an agonist and said individual is in need of prevention of or treatment for a neuronal cell death-related disorder. In certain embodiments, said neuronal cell death-related disorder is selected from the group consisting of:

(a) Alzheimer's disease; (b) Parkinson's disease; (c) stroke; (d) motor-neuron disease; (e) learning or memory impairment; (f) traumatic brain injury; (g) spinal cord injury; (h) peripheral neuropathy; and (i) prion-associated disease.

In some embodiments, Alzheimer's disease encompasses mild cognitive impairment (MCI).

Motor-neuron disease includes, but is not limited to, amyotrophic lateral sclerosis, progressive spinal muscular atrophy, progressive bulbar palsy, and primary lateral sclerosis. Peripheral neuropathy includes, but is not limited, to diabetic neuropathy and neuropathy involving the sciatic nerve. Diabetic neuropathy includes, but is not limited to, distal symmetrical polyneuropathy (DSP).

In some embodiments, the neuronal cell death-related disorder is Alzheimer's disease. In some embodiments, the neuronal cell death-related disorder is Parkinson's disease. In some embodiments, the neuronal cell death-related disorder is stroke. In some embodiments, the neuronal cell death-related disorder is motor-neuron disease. In some embodiments, the neuronal cell death-related disorder is learning or memory impairment. In some embodiments, the neuronal cell death-related disorder is traumatic brain injury. In some embodiments, the neuronal cell death-related disorder is spinal cord injury. In some embodiments, the neuronal cell death-related disorder is peripheral neuropathy. In some embodiments, the neuronal cell death-related disorder is prion-associated disease.

In certain embodiments, said modulator is an inverse agonist or an antagonist and said individual is in need of opposition to protection from neuronal cell death.

In certain embodiments, said modulator is an inverse agonist or an antagonist and said individual is in need of prevention of or treatment for a neuronal cell proliferative disorder. In certain embodiments, said neuronal cell proliferative disorder is neuroblastoma. In other embodiments, said neuronal cell proliferative disorder is medulloblastoma.

In certain embodiments, said modulator is an agonist and said individual is in need of reducing muscle cell death.

In certain embodiments, said modulator is an agonist and said individual is in need of prevention of or treatment for a muscle cell death-related disorder. In certain embodiments, said muscle cell death-related disorder is selected from the group consisting of:

(a) cerebral amyloid beta-protein angiopathy; (b) myocardial infarction; and (c) congestive heart failure.

In some embodiments, the muscle cell death-related disorder is cerebral amyloid beta-protein angiopathy. In some embodiments, the muscle cell death-related disorder is myocardial infarction. In some embodiments, the muscle cell death-related disorder is congestive heart failure.

In certain embodiments, said modulator is an inverse agonist or an antagonist and said individual is in need of opposition to protection from muscle cell death.

In certain embodiments, said modulator is an inverse agonist or an antagonist and said individual is in need of prevention of or treatment for a muscle cell proliferative disorder. In certain embodiments, said muscle cell proliferative disorder is selected from the group consisting of atherosclerosis, restenosis, and tumor-supportive angiogenesis.

In a seventh aspect, the invention features the method of the sixth aspect, wherein an in vitro cell culture comprises one or more cells comprising said Humanin GPCR, and wherein said cell culture is in need of prevention of or treatment for cell death.

In certain embodiments, said method comprises administration of an effective amount of the modulator to a cell comprising the receptor.

In certain embodiments, said modulator is an agonist.

In certain embodiments, said one or more cells comprises one or more neuronal cells.

In certain embodiments, said one or more cells comprises one or more muscle cells.

In some embodiments, said one or more cells comprises one or more cells that are non-neuronal and non-muscle.

In certain embodiments, culture medium used in said cell culture comprises said modulator.

In certain embodiments, said modulator is provided as a supplement to cell culture medium.

In certain embodiments, said modulator is identified according to a method of the first aspect.

In certain embodiments, said modulator is a compound according to the second aspect.

In certain embodiments, said modulator is a compound according to the third aspect.

In certain embodiments, said modulator is Compound 1.

In certain embodiments, said modulator is Compound 1 and said effective amount of Compound 1 is about 0.5 μM, about 1.0 μM, about 2.0 μM, about 3.0 μM, about 4.0 μM, about 5.0 μM, about 6.0 μM, about 7.0 μM, about 8.0 μM, about 9.0 μM, about 10.0 μM, about 20.0 μM, about 30.0 μM, about 40.0 μM or about 50 μM.

In an eighth aspect, the invention features a method of modulating the activity of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for reducing neuronal cell death in an individual in need of said modulation, comprising contacting said receptor with a therapeutically effective amount of a modulator of the receptor.

The invention also features a method of modulating the activity of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for preventing or treating a neuronal cell death-related disorder in an individual in need of said modulation, comprising contacting said receptor with a therapeutically effective amount of a modulator of the receptor. In certain embodiments, said neuronal cell death-related disorder is selected from the group consisting of:

(a) Alzheimer's disease; (b) Parkinson's disease; (c) stroke; (d) motor-neuron disease; (e) learning or memory impairment; (d) traumatic brain injury; (e) spinal cord injury; (f) peripheral neuropathy; and (g) prion-associated disease.

In some embodiments, the neuronal cell death-related disorder is Alzheimer's disease. In some embodiments, the neuronal cell death-related disorder is Parkinson's disease. In some embodiments, the neuronal cell death-related disorder is stroke. In some embodiments, the neuronal cell death-related disorder is motor-neuron disease. In some embodiments, the neuronal cell death-related disorder is learning or memory impairment. In some embodiments, the neuronal cell death-related disorder is traumatic brain injury. In some embodiments, the neuronal cell death-related disorder is spinal cord injury. In some embodiments, the neuronal cell death-related disorder is peripheral neuropathy. In some embodiments, the neuronal cell death-related disorder is prion-associated disease.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist. In certain embodiments, said agonist is a compound according to the second aspect. In certain embodiments, said agonist is a compound according to the third aspect.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is neuroprotective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC 50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

In certain embodiments, said contacting comprises oral administration of said modulator to said individual.

In some embodiments, Alzheimer's disease encompasses mild cognitive impairment (MCI).

Motor-neuron disease includes, but is not limited to, amyotrophic lateral sclerosis, progressive spinal muscular atrophy, progressive bulbar palsy, and primary lateral sclerosis. Peripheral neuropathy includes, but is not limited, to diabetic neuropathy and neuropathy involving the sciatic nerve. Diabetic neuropathy includes, but is not limited to, distal symmetrical polyneuropathy (DSP).

The invention also relates to a method of modulating a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for preventing or treating a neuronal cell proliferative disorder in an individual in need of said modulation, comprising contacting a therapeutically effective amount of a modulator of said receptor with said receptor. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist. In certain embodiments, said neuronal cell proliferative disorder is neuroblastoma. In other embodiments, said neuronal cell proliferative disorder is medulloblastoma.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In an ninth aspect, the invention features a method of modulating the activity of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for reducing muscle cell death in an individual in need of said modulation, comprising contacting said receptor with a therapeutically effective amount of a modulator of the receptor.

The invention also features a method of modulating the activity of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for preventing or treating a muscle cell death-related disorder in an individual in need of said modulation, comprising contacting said receptor with a therapeutically effective amount of a modulator of the receptor. In certain embodiments, said muscle cell death-related disorder is selected from the group consisting of:

(a) cerebral amyloid beta-protein angiopathy; (b) myocardial infarction; and (c) congestive heart failure.

In some embodiments, the muscle cell death-related disorder is cerebral amyloid beta-protein angiopathy. In some embodiments, the muscle cell death-related disorder is myocardial infarction. In some embodiments, the muscle cell death-related disorder is congestive heart failure.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist. In certain embodiments, said agonist is a compound according to the second aspect. In certain embodiments, said agonist is a compound according to the third aspect.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is myoprotective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

In certain embodiments, said contacting comprises oral administration of said modulator to said individual.

The invention also relates to a method of modulating a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for preventing or treating a muscle cell proliferative disorder in an individual in need of said modulation, comprising contacting a therapeutically effective amount of a modulator of said receptor with said receptor. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist. In certain embodiments, said muscle cell proliferative disorder is selected from the group consisting of:

(a) atherosclerosis; (b) restenosis; and (c) tumor-supportive angiogenesis.

In some embodiments, the muscle cell proliferative disorder is atherosclerosis. In some embodiments, the muscle cell proliferative disorder is restenosis. In some embodiments, the muscle cell proliferative disorder is tumor-supportive angiogenesis.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a tenth aspect, the invention features a method of modulating the activity of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for reducing cell death in an individual in need of said modulation, wherein said cell in not neuronal and not muscle, comprising contacting said receptor with a therapeutically effective amount of a modulator of the receptor.

The invention also features a method of modulating the activity of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for preventing or treating a cell death-related disorder in an individual in need of said modulation, comprising contacting said receptor with a therapeutically effective amount of a modulator of the receptor. In certain embodiments, said cell resides in or derives from a tissue that expresses human FPRL2. In certain embodiments, said tissue expressing human FPRL2 is selected from the group consisting of sciatic nerve, anterior hippocampus, whole brain, hippocampus, substantia nigra, spleen, heart, lung, pancreas, bone, and ovary.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist. In certain embodiments, said agonist is a compound according to the second aspect. In certain embodiments, said agonist is a compound according to the third aspect.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is cell death-protective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC 50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

In certain embodiments, said contacting comprises oral administration of said modulator to said individual.

The invention also relates to a method of modulating a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for preventing or treating a cell proliferative disorder in an individual in need of said modulation, comprising contacting a therapeutically effective amount of a modulator of said receptor with said receptor. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In an eleventh aspect, the invention features a method of reducing neuronal cell death in an individual in need of said reducing, comprising contacting a therapeutically effective amount of a modulator of a Humanin GPCR with said receptor, said GPCR comprising an FPRL2 amino acid sequence.

The invention also features a method of preventing or treating a neuronal cell death-related disorder in an individual in need of said prevention or treatment, comprising contacting a therapeutically effective amount of a modulator of a Human GPCR with said receptor, said receptor comprising an FPRL2 amino acid sequence. In certain embodiments, said neuronal cell death-related disorder is selected from the group consisting of:

(a) Alzheimer's disease; (b) Parkinson's disease; (c) stroke; (d) motor-neuron disease; (e) learning or memory impairment; (d) traumatic brain injury; (e) spinal cord injury; (f) peripheral neuropathy; and (g) prion-associated disease.

In some embodiments, the neuronal cell death-related disorder is Alzheimer's disease. In some embodiments, the neuronal cell death-related disorder is Parkinson's disease. In some embodiments, the neuronal cell death-related disorder is stroke. In some embodiments, the neuronal cell death-related disorder is motor-neuron disease. In some embodiments, the neuronal cell death-related disorder is learning or memory impairment. In some embodiments, the neuronal cell death-related disorder is traumatic brain injury. In some embodiments, the neuronal cell death-related disorder is spinal cord injury. In some embodiments, the neuronal cell death-related disorder is peripheral neuropathy. In some embodiments, the neuronal cell death-related disorder is prion-associated disease.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist. In certain embodiments, said agonist is a compound according to the second aspect. In certain embodiments, said agonist is a compound according to the third aspect.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is neuroprotective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

In certain embodiments, said contacting comprises oral administration of said modulator to said individual.

In some embodiments, Alzheimer's disease encompasses mild cognitive impairment (MCI).

Motor-neuron disease includes, but is not limited to, amyotrophic lateral sclerosis, progressive spinal muscular atrophy, progressive bulbar palsy, and primary lateral sclerosis. Peripheral neuropathy includes, but is not limited, to diabetic neuropathy and neuropathy involving the sciatic nerve. Diabetic neuropathy includes, but is not limited to, distal symmetrical polyneuropathy (DSP).

The invention also features a method of preventing or treating a neuronal cell proliferative disorder in an individual in need of said prevention or treatment, comprising contacting a therapeutically effective amount of a modulator of a Humanin GPCR with said receptor, said receptor comprising an FPRL2 amino acid sequence. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist. In certain embodiments, said neuronal cell proliferative disorder is neuroblastoma. In other embodiments, said neuronal cell proliferative disorder is medulloblastoma.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In an twelfth aspect, the invention features a method of reducing muscle cell death in an individual in need of said reducing, comprising contacting a therapeutically effective amount of a modulator of a Humanin GPCR with said receptor, said GPCR comprising an FPRL2 amino acid sequence.

The invention also features a method of preventing or treating a muscle cell death-related disorder in an individual in need of said prevention or treatment, comprising contacting a therapeutically effective amount of a modulator of a Humanin GPCR with said receptor, said receptor comprising an FPRL2 amino acid sequence. In certain embodiments, said muscle cell death-related disorder is selected from the group consisting of:

(a) cerebral amyloid beta-protein angiopathy; (b) myocardial infarction; and (c) congestive heart failure.

In some embodiments, the muscle cell death-related disorder is cerebral amyloid beta-protein angiopathy. In some embodiments, the muscle cell death-related disorder is myocardial infarction. In some embodiments, the muscle cell death-related disorder is congestive heart failure.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist. In certain embodiments, said agonist is a compound according to the second aspect. In certain embodiments, said agonist is a compound according to the third aspect.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is myoprotective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

In certain embodiments, said contacting comprises oral administration of said modulator to said individual.

The invention also relates to a method of preventing or treating a muscle cell proliferative disorder in an individual in need of said prevention or treatment, comprising contacting a therapeutically effective amount of a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, with the receptor. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist. In certain embodiments, said muscle cell proliferative disorder is selected from the group consisting of:

(a) atherosclerosis;

(b) restenosis; and

(c) tumor-supportive angiogenesis.

In some embodiments, the muscle cell proliferative disorder is atherosclerosis. In some embodiments, the muscle cell proliferative disorder is restenosis. In some embodiments, the muscle cell proliferative disorder is tumor-supportive angiogenesis.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a thirteenth aspect, the invention features a method of reducing cell death in an individual in need of said reducing, comprising contacting a therapeutically effective amount of a modulator of a Humanin GPCR with said receptor, said GPCR comprising an FPRL2 amino acid sequence.

The invention also features a method of preventing or treating a cell death-related disorder in an individual in need of said prevention or treatment, comprising contacting a therapeutically effective amount of a modulator of a Humanin GPCR with said receptor, said receptor comprising an FPRL2 amino acid sequence. In certain embodiments, said cell resides in or derives from a tissue that expresses human FPRL2. In certain embodiments, said tissue expressing human FPRL2 is selected from the group consisting of sciatic nerve, anterior hippocampus, whole brain, hippocampus, substantia nigra, spleen, heart, lung, pancreas, bone, and ovary.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist. In certain embodiments, said agonist is a compound according to the second aspect. In certain embodiments, said agonist is a compound according to the third aspect.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is cell death-protective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

In certain embodiments, said contacting comprises oral administration of said modulator to said individual.

The invention also relates to a method of preventing or treating cell proliferative disorder in an individual in need of said prevention or treatment, comprising contacting a therapeutically effective amount of a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, with the receptor. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a fourteenth aspect, the invention features a pharmaceutical or physiologically acceptable composition comprising, consisting essentially of, or consisting of a modulator a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist. In certain embodiments, said agonist is a compound according to the second aspect. In certain embodiments, said agonist is a compound according to the third aspect.

In certain embodiments, said composition is pharmaceutical. In certain embodiments, said composition is physiologically acceptable.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is cell death-protective.

In certain embodiments, said modulator is neuroprotective.

In certain embodiments, said modulator is myoprotective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

In a fifteenth aspect, the invention features a method of reducing neuronal cell death comprising providing or administering to an individual in need of said reduction said pharmaceutical or physiologically acceptable composition of the fourteenth aspect.

The invention also features a method of preventing or treating a neuronal cell death-related disorder comprising providing or administering to an individual in need of said prevention or treatment said pharmaceutical or physiologically acceptable composition of the fourteenth aspect. In certain embodiments, said neuronal cell death-related disorder is selected from the group consisting of:

(a) Alzheimer's disease; (b) Parkinson's disease; (c) stroke; (d) motor-neuron disease; (e) learning or memory impairment; (f) traumatic brain injury; (g) spinal cord injury; (h) peripheral neuropathy; and (i) prion-associated disease.

In some embodiments, Alzheimer's disease encompasses mild cognitive impairment (MCI).

Motor-neuron disease includes, but is not limited to, amyotrophic lateral sclerosis, progressive spinal muscular atrophy, progressive bulbar palsy, and primary lateral sclerosis. Peripheral neuropathy includes, but is not limited, to diabetic neuropathy and neuropathy involving the sciatic nerve. Diabetic neuropathy includes, but is not limited to, distal symmetrical polyneuropathy (DSP).

In some embodiments, the neuronal cell death-related disorder is Alzheimer's disease. In some embodiments, the neuronal cell death-related disorder is Parkinson's disease. In some embodiments, the neuronal cell death-related disorder is stroke. In some embodiments, the neuronal cell death-related disorder is motor-neuron disease. In some embodiments, the neuronal cell death-related disorder is learning or memory impairment. In some embodiments, the neuronal cell death-related disorder is traumatic brain injury. In some embodiments, the neuronal cell death-related disorder is spinal cord injury. In some embodiments, the neuronal cell death-related disorder is peripheral neuropathy. In some embodiments, the neuronal cell death-related disorder is prion-associated disease.

In certain embodiments, said modulator is an agonist.

In certain embodiments, a therapeutically effective amount of said pharmaceutical or physiologically acceptable composition is provided or administered to said individual.

In certain embodiments, said providing or administering of said pharmaceutical or physiologically acceptable composition is oral.

The invention also relates to a method of preventing or treating a neuronal cell proliferative disorder comprising providing or administering to an individual in need of said prevention or treatment said pharmaceutical or physiologically acceptable composition of the fourteenth aspect. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist. In certain embodiments, said neuronal cell proliferative disorder is neuroblastoma. In other embodiments, said neuronal cell proliferative disorder is medulloblastoma.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a sixteenth aspect, the invention features a method of reducing muscle cell death comprising providing or administering to an individual in need of said reduction said pharmaceutical or physiologically acceptable composition of the fourteenth aspect.

The invention also features a method of preventing or treating a muscle cell death-related disorder comprising providing or administering to an individual in need of said prevention or treatment said pharmaceutical or physiologically acceptable composition of the fourteenth aspect. In certain embodiments, said cell death-related disorder is selected from the group consisting of:

(a) cerebral amyloid beta-protein angiopathy;

(b) myocardial infarction; and

(c) congestive heart failure.

In some embodiments, the muscle cell death-related disorder is cerebral amyloid beta-protein angiopathy. In some embodiments, the muscle cell death-related disorder is myocardial infarction. In some embodiments, the muscle cell death-related disorder is congestive heart failure.

In certain embodiments, said modulator is an agonist.

In certain embodiments, a therapeutically effective amount of said pharmaceutical or physiologically acceptable composition is provided or administered to said individual.

In certain embodiments, said providing or administering of said pharmaceutical or physiologically acceptable composition is oral.

The invention also relates to a method of preventing or treating a muscle cell proliferative disorder comprising providing or administering to an individual in need of said prevention or treatment said pharmaceutical or physiologically acceptable composition of the fourteenth aspect. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist. In certain embodiments, said muscle cell proliferative disorder selected from the group consisting of:

(a) atherosclerosis;

(b) restenosis; and

(c) tumor-supportive angiogenesis.

In some embodiments, the muscle cell proliferative disorder is atherosclerosis. In some embodiments, the muscle cell proliferative disorder is restenosis. In some embodiments, the muscle cell proliferative disorder is tumor-supportive angiogenesis.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a seventeenth aspect, the invention features a method of reducing cell death comprising providing or administering to an individual in need of said reduction said pharmaceutical or physiologically acceptable composition of the fourteenth aspect.

The invention also features a method of preventing or treating a cell death-related disorder comprising providing or administering to an individual in need of said prevention or treatment said pharmaceutical or physiologically acceptable composition of the fourteenth aspect. In certain embodiments, said cell resides in or derives from a tissue that expresses human FPRL2. In certain embodiments, said tissue expressing human FPRL2 is selected from the group consisting of sciatic nerve, anterior hippocampus, whole brain, hippocampus, substantia nigra, spleen, heart, lung, pancreas, bone, and ovary.

In certain embodiments, a therapeutically effective amount of said pharmaceutical or physiologically acceptable composition is provided or administered to said individual.

In certain embodiments, said providing or administering of said pharmaceutical or physiologically acceptable composition is oral.

The invention also relates to a method of preventing or treating a cell proliferative disorder comprising providing or administering to an individual in need of said prevention or treatment said pharmaceutical or physiologically acceptable composition of the fourteenth aspect. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In an eighteenth aspect, the invention features a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for use in a method of treatment of the human animal body by therapy.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist. In certain embodiments, said agonist is a compound according to the second aspect. In certain embodiments, said agonist is a compound according to the third aspect.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC 50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

In certain embodiments, said animal is a mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, or non-human primate. More preferred of human or animal is human.

In an nineteenth aspect, the invention features a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for use in a method of reducing neuronal cell death in the human animal body by therapy.

The invention also features a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for use in a method of prevention of or treatment for a neuronal cell death-related disorder in a human or animal body by therapy. In certain embodiments, said neuronal cell death-related disorder is selected from the group consisting of:

(a) Alzheimer's disease;

(b) Parkinson's disease;

(c) stroke;

(d) motor-neuron disease;

(e) learning or memory impairment;

(f) traumatic brain injury;

(g) spinal cord injury;

(h) peripheral neuropathy; and

(i) prion-associated disease.

In some embodiments, Alzheimer's disease encompasses mild cognitive impairment (MCI).

Motor-neuron disease includes, but is not limited to, amyotrophic lateral sclerosis, progressive spinal muscular atrophy, progressive bulbar palsy, and primary lateral sclerosis. Peripheral neuropathy includes, but is not limited, to diabetic neuropathy and neuropathy involving the sciatic nerve. Diabetic neuropathy includes, but is not limited to, distal symmetrical polyneuropathy (DSP).

In some embodiments, the neuronal cell death-related disorder is Alzheimer's disease. In some embodiments, the neuronal cell death-related disorder is Parkinson's disease. In some embodiments, the neuronal cell death-related disorder is stroke. In some embodiments, the neuronal cell death-related disorder is motor-neuron disease. In some embodiments, the neuronal cell death-related disorder is learning or memory impairment. In some embodiments, the neuronal cell death-related disorder is traumatic brain injury. In some embodiments, the neuronal cell death-related disorder is spinal cord injury. In some embodiments, the neuronal cell death-related disorder is peripheral neuropathy. In some embodiments, the neuronal cell death-related disorder is prion-associated disease.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist. In certain embodiments, said agonist is a compound according to the second aspect. In certain embodiments, said agonist is a compound according to the third aspect.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is neuroprotective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ HD NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

The invention also relates to a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for use in a method of prevention of or treatment for a neuronal cell proliferative disorder in a human or animal body by therapy. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist. In certain embodiments, said neuronal cell proliferative disorder is neuroblastoma. In other embodiments, said neuronal cell proliferative disorder is medulloblastoma.

In certain embodiments, said animal is a mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, or non-human primate. More preferred of human or animal is human.

In an twentieth aspect, the invention features a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for use in a method of reducing muscle cell death in the human animal body by therapy.

The invention also features a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for use in a method of prevention of or treatment for a muscle cell death-related disorder in a human or animal body by therapy. In certain embodiments, said muscle cell death-related disorder is selected from the group consisting of:

(a) cerebral amyloid beta-protein angiopathy;

(b) myocardial infarction; and

(c) congestive heart failure.

In some embodiments, the muscle cell death-related disorder is cerebral amyloid beta-protein angiopathy. In some embodiments, the muscle cell death-related disorder is myocardial infarction. In some embodiments, the muscle cell death-related disorder is congestive heart failure.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist. In certain embodiments, said agonist is a compound according to the second aspect. In certain embodiments, said agonist is a compound according to the third aspect.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is myoprotective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

The invention also relates to a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for use in a method of prevention of or treatment for a muscle cell proliferative disorder in a human or animal body by therapy. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist. In certain embodiments, said muscle cell proliferative disorder selected from the group consisting of:

(a) atherosclerosis;

(b) restenosis; and

(c) tumor-supportive angiogenesis.

In some embodiments, the muscle cell proliferative disorder is atherosclerosis. In some embodiments, the muscle cell proliferative disorder is restenosis. In some embodiments, the muscle cell proliferative disorder is tumor-supportive angiogenesis.

In certain embodiments, said animal is a mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, or non-human primate. More preferred of human or animal is human.

In a twenty-first aspect, the invention features a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for use in a method of reducing cell death in the human animal body by therapy.

The invention also features a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for use in a method of prevention of or treatment for a cell death-related disorder in a human or animal body by therapy. In certain embodiments, said cell resides in or derives from a tissue that expresses human FPRL2. In certain embodiments, said tissue expressing human FPRL2 is selected from the group consisting of sciatic nerve, anterior hippocampus, whole brain, hippocampus, substantia nigra, spleen, heart, lung, pancreas, bone, and ovary.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist. In certain embodiments, said agonist is a compound according to the second aspect. In certain embodiments, said agonist is a compound according to the third aspect.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is cell death-protective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

The invention also relates to a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for use in a method of prevention of or treatment for a cell proliferative disorder in a human or animal body by therapy. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist.

In certain embodiments, said animal is a mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, or non-human primate. More preferred of human or animal is human.

In a twenty-second aspect, the invention features a method of using a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for the preparation of a medicament for the reduction of neuronal cell death.

The invention also features a method of using a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for the preparation of a medicament for the prevention or treatment of a neuronal cell death-related disorder. In certain embodiments, said neuronal cell death-related disorder is selected from the group consisting of:

(a) Alzheimer's disease; (b) Parkinson's disease; (c) stroke; (d) motor-neuron disease; (e) learning or memory impairment; (d) traumatic brain injury; (e) spinal cord injury; (f) peripheral neuropathy; and (g) prion-associated disease.

In some embodiments, the neuronal cell death-related disorder is Alzheimer's disease. In some embodiments, the neuronal cell death-related disorder is Parkinson's disease. In some embodiments, the neuronal cell death-related disorder is stroke. In some embodiments, the neuronal cell death-related disorder is motor-neuron disease. In some embodiments, the neuronal cell death-related disorder is learning or memory impairment. In some embodiments, the neuronal cell death-related disorder is traumatic brain injury. In some embodiments, the neuronal cell death-related disorder is spinal cord injury. In some embodiments, the neuronal cell death-related disorder is peripheral neuropathy. In some embodiments, the neuronal cell death-related disorder is prion-associated disease.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist. In certain embodiments, said agonist is a compound according to the second aspect. In certain embodiments, said agonist is a compound according to the third aspect.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is neuroprotective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

In some embodiments, Alzheimer's disease encompasses mild cognitive impairment (MCI).

Motor-neuron disease includes, but is not limited to, amyotrophic lateral sclerosis, progressive spinal muscular atrophy, progressive bulbar palsy, and primary lateral sclerosis. Peripheral neuropathy includes, but is not limited, to diabetic neuropathy and neuropathy involving the sciatic nerve. Diabetic neuropathy includes, but is not limited to, distal symmetrical polyneuropathy (DSP).

The invention also features a method of using a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for the preparation of a medicament for preventing or treating a neuronal cell proliferative disorder. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist. In certain embodiments, said neuronal cell proliferative disorder is neuroblastoma. In other embodiments, said neuronal cell proliferative disorder is medulloblastoma.

In a twenty-third aspect, the invention features a method of using a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for the preparation of a medicament for the reduction of muscle cell death.

The invention also features a method of using a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for the preparation of a medicament for the prevention or treatment of a muscle cell death-related disorder. In certain embodiments, said muscle cell death-related disorder is selected from the group consisting of:

(a) cerebral amyloid beta-protein angiopathy; (b) myocardial infarction; and (c) congestive heart failure.

In some embodiments, the muscle cell death-related disorder is cerebral amyloid beta-protein angiopathy. In some embodiments, the muscle cell death-related disorder is myocardial infarction. In some embodiments, the muscle cell death-related disorder is congestive heart failure.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist. In certain embodiments, said agonist is a compound according to the second aspect. In certain embodiments, said agonist is a compound according to the third aspect.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is myoprotective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

The invention also relates to a method of using a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for the preparation of a medicament for the prevention or treatment of a muscle cell proliferative disorder. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist. In certain embodiments, said muscle cell proliferative disorder is selected from the group consisting of:

(a) atherosclerosis;

(b) restenosis; and

(c) tumor-supportive angiogenesis.

In some embodiments, the muscle cell proliferative disorder is atherosclerosis. In some embodiments, the muscle cell proliferative disorder is restenosis. In some embodiments, the muscle cell proliferative disorder is tumor-supportive angiogenesis.

In a twenty-fourth aspect, the invention features a method of using a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for the preparation of a medicament for the reduction of cell death.

The invention also features a method of using a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for the preparation of a medicament for the prevention or treatment of a cell death-related disorder. In certain embodiments, said cell resides in or derives from a tissue that expresses human FPRL2. In certain embodiments, said tissue expressing human FPRL2 is selected from the group consisting of sciatic nerve, anterior hippocampus, whole brain, hippocampus, substantia nigra, spleen, heart, lung, pancreas, bone, and ovary.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist. In certain embodiments, said agonist is a compound according to the second aspect. In certain embodiments, said agonist is a compound according to the third aspect.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is cell death-protective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

The invention also relates to a method of using a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for the preparation of a medicament for the prevention or treatment of a cell proliferative disorder. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist.

In a twenty-fifth aspect, the invention features a method of modulating the activity of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for reducing neuronal cell death in an individual in need of said modulation, comprising contacting said receptor with a therapeutically effective amount of a modulator of the receptor. In certain embodiments, said method comprises performing a method according to the first aspect to thereby identify a modulator.

The invention also features a method of modulating the activity of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for preventing or treating a neuronal cell death-related disorder in an individual in need of said modulation, comprising contacting said receptor with a therapeutically effective amount of a modulator of the receptor. In certain embodiments, said method comprises performing a method according to the first aspect to thereby identify a modulator. In certain embodiments, said neuronal cell death-related disorder is selected from the group consisting of:

(a) Alzheimer's disease; (b) Parkinson's disease; (c) stroke; (d) motor-neuron disease; (e) learning or memory impairment; (d) traumatic brain injury; (e) spinal cord injury; (f) peripheral neuropathy; and (g) prion-associated disease.

In some embodiments, the neuronal cell death-related disorder is Alzheimer's disease. In some embodiments, the neuronal cell death-related disorder is Parkinson's disease. In some embodiments, the neuronal cell death-related disorder is stroke. In some embodiments, the neuronal cell death-related disorder is motor-neuron disease. In some embodiments, the neuronal cell death-related disorder is learning or memory impairment. In some embodiments, the neuronal cell death-related disorder is traumatic brain injury. In some embodiments, the neuronal cell death-related disorder is spinal cord injury. In some embodiments, the neuronal cell death-related disorder is peripheral neuropathy. In some embodiments, the neuronal cell death-related disorder is prion-associated disease.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is according to the fourth aspect. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is neuroprotective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM less than 3 μM, less than 2 μM, or less than 1 μM.

In certain embodiments, said contacting comprises oral administration of said modulator to said individual.

In some embodiments, Alzheimer's disease encompasses mild cognitive impairment (MCI).

Motor-neuron disease includes, but is not limited to, amyotrophic lateral sclerosis, progressive spinal muscular atrophy, progressive bulbar palsy, and primary lateral sclerosis. Peripheral neuropathy includes, but is not limited, to diabetic neuropathy and neuropathy involving the sciatic nerve. Diabetic neuropathy includes, but is not limited to, distal symmetrical polyneuropathy (DSP).

The invention also relates to a method of modulating a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for preventing or treating a neuronal cell proliferative disorder in an individual in need of said modulation, comprising contacting a therapeutically effective amount of a modulator of said receptor with said receptor. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist. In certain embodiments, said neuronal cell proliferative disorder is neuroblastoma. In other embodiments, said neuronal cell proliferative disorder is medulloblastoma.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a twenty-sixth aspect, the invention features a method of modulating the activity of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for reducing muscle cell death in an individual in need of said modulation, comprising contacting said receptor with a therapeutically effective amount of a modulator of the receptor. In certain embodiments, said method comprises performing a method according to the first aspect to thereby identify a modulator.

The invention also features a method of modulating the activity of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for preventing or treating a muscle cell death-related disorder in an individual in need of said modulation, comprising contacting said receptor with a therapeutically effective amount of a modulator of the receptor. In certain embodiments, said method comprises performing a method according to the first aspect to thereby identify a modulator. In certain embodiments, said muscle cell death-related disorder is selected from the group consisting of:

(a) cerebral amyloid beta-protein angiopathy; (b) myocardial infarction; and (c) congestive heart failure.

In some embodiments, the muscle cell death-related disorder is cerebral amyloid beta-protein angiopathy. In some embodiments, the muscle cell death-related disorder is myocardial infarction. In some embodiments, the muscle cell death-related disorder is congestive heart failure.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is according to the fourth aspect. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is myoprotective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

In certain embodiments, said contacting comprises oral administration of said modulator to said individual.

The invention also relates to a method of modulating a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for preventing or treating a muscle cell proliferative disorder in an individual in need of said modulation, comprising contacting a therapeutically effective amount of a modulator of said receptor with said receptor. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist. In certain embodiments, said muscle cell proliferative disorder is selected from the group consisting of:

(a) atherosclerosis;

(b) restenosis; and

(c) tumor-supportive angiogenesis.

In some embodiments, the muscle cell proliferative disorder is atherosclerosis. In some embodiments, the muscle cell proliferative disorder is restenosis. In some embodiments, the muscle cell proliferative disorder is tumor-supportive angiogenesis.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a twenty-seventh aspect, the invention features a method of modulating the activity of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for reducing cell death in an individual in need of said modulation, wherein said cell in not neuronal and not muscle, comprising contacting said receptor with a therapeutically effective amount of a modulator of the receptor. In certain embodiments, said method comprises performing a method according to the first aspect to thereby identify a modulator.

The invention also features a method of modulating the activity of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for preventing or treating a cell death-related disorder in an individual in need of said modulation, comprising contacting said receptor with a therapeutically effective amount of a modulator of the receptor. In certain embodiments, said method comprises performing a method according to the first aspect to thereby identify a modulator. In certain embodiments, said cell resides in or derives from a tissue that expresses human FPRL2. In certain embodiments, said tissue expressing human FPRL2 is selected from the group consisting of sciatic nerve, anterior hippocampus, whole brain, hippocampus, substantia nigra, spleen, heart, lung, pancreas, bone, and ovary.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is according to the fourth aspect. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is cell death-protective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of Less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

In certain embodiments, said contacting comprises oral administration of said modulator to said individual.

The invention also relates to a method of modulating a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for preventing or treating a cell proliferative disorder in an individual in need of said modulation, comprising contacting a therapeutically effective amount of a modulator of said receptor with said receptor. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In an twenty-eighth aspect, the invention features a method of reducing neuronal cell death in an individual in need of said reducing, comprising contacting a therapeutically effective amount of a modulator of a Humanin GPCR with said receptor, said GPCR comprising an FPRL2 amino acid sequence. In certain embodiments, said method comprises performing a method according to the first aspect to thereby identify a modulator.

The invention also features a method of preventing or treating a neuronal cell death-related disorder in an individual in need of said prevention or treatment, comprising contacting a therapeutically effective amount of a modulator of a Human GPCR with said receptor, said receptor comprising an FPRL2 amino acid sequence. In certain embodiments, said method comprises performing a method according to the first aspect to thereby identify a modulator. In certain embodiments, said neuronal cell death-related disorder is selected from the group consisting of:

(a) Alzheimer's disease; (b) Parkinson's disease; (c) stroke; (d) motor-neuron disease; (e) learning or memory impairment; (d) traumatic brain injury; (e) spinal cord injury; (i) peripheral neuropathy; and (g) prion-associated disease.

In some embodiments, the neuronal cell death-related disorder is Alzheimer's disease. In some embodiments, the neuronal cell death-related disorder is Parkinson's disease. In some embodiments, the neuronal cell death-related disorder is stroke. In some embodiments, the neuronal cell death-related disorder is motor-neuron disease. In some embodiments, the neuronal cell death-related disorder is learning or memory impairment. In some embodiments, the neuronal cell death-related disorder is traumatic brain injury. In some embodiments, the neuronal cell death-related disorder is spinal cord injury. In some embodiments, the neuronal cell death-related disorder is peripheral neuropathy. In some embodiments, the neuronal cell death-related disorder is prion-associated disease.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is according to the fourth aspect. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is neuroprotective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

In certain embodiments, said contacting comprises oral administration of said modulator to said individual.

In some embodiments, Alzheimer's disease encompasses mild cognitive impairment (MCI).

Motor-neuron disease includes, but is not limited to, amyotrophic lateral sclerosis, progressive spinal muscular atrophy, progressive bulbar palsy, and primary lateral sclerosis. Peripheral neuropathy includes, but is not limited, to diabetic neuropathy and neuropathy involving the sciatic nerve. Diabetic neuropathy includes, but is not limited to, distal symmetrical polyneuropathy (DSP).

The invention also features a method of preventing or treating a neuronal cell proliferative disorder in an individual in need of said prevention or treatment, comprising contacting a therapeutically effective amount of a modulator of a Humanin GPCR with said receptor, said receptor comprising an FPRL2 amino acid sequence. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist. In certain embodiments, said neuronal cell proliferative disorder is neuroblastoma. In other embodiments, said neuronal cell proliferative disorder is medulloblastoma.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In an twenty-ninth aspect, the invention features a method of reducing muscle cell death in an individual in need of said reducing, comprising contacting a therapeutically effective amount of a modulator of a Humanin GPCR with said receptor, said GPCR comprising an FPRL2 amino acid sequence. In certain embodiments, said method comprises performing a method according to the first aspect to thereby identify a modulator.

The invention also features a method of preventing or treating a muscle cell death-related disorder in an individual in need of said prevention or treatment, comprising contacting a therapeutically effective amount of a modulator of a Humanin GPCR with said receptor, said receptor comprising an FPRL2 amino acid sequence. In certain embodiments, said method comprises performing a method according to the first aspect to thereby identify a modulator. In certain embodiments, said muscle cell death-related disorder is selected from the group consisting of:

(a) cerebral amyloid beta-protein angiopathy; (b) myocardial infarction; and (c) congestive heart failure.

In some embodiments, the muscle cell death-related disorder is cerebral amyloid beta-protein angiopathy. In some embodiments, the muscle cell death-related disorder is myocardial infarction. In some embodiments, the muscle cell death-related disorder is congestive heart failure.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is according to the fourth aspect. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is myoprotective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

In certain embodiments, said contacting comprises oral administration of said modulator to said individual.

The invention also relates to a method of preventing or treating a muscle cell proliferative disorder in an individual in need of said prevention or treatment, comprising contacting a therapeutically effective amount of a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, with the receptor. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist. In certain embodiments, said muscle cell proliferative disorder is selected from the group consisting of:

(a) atherosclerosis;

(b) restenosis; and

(c) tumor-supportive angiogenesis.

In some embodiments, the muscle cell proliferative disorder is atherosclerosis. In some embodiments, the muscle cell proliferative disorder is restenosis. In some embodiments, the muscle cell proliferative disorder is tumor-supportive angiogenesis.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a thirtieth aspect, the invention features a method of reducing cell death in an individual in need of said reducing, comprising contacting a therapeutically effective amount of a modulator of a Humanin GPCR with said receptor, said GPCR comprising an FPRL2 amino acid sequence. In certain embodiments, said method comprises performing a method according to the first aspect to thereby identify a modulator.

The invention also features a method of preventing or treating a cell death-related disorder in an individual in need of said prevention or treatment, comprising contacting a therapeutically effective amount of a modulator of a Humanin GPCR with said receptor, said receptor comprising an FPRL2 amino acid sequence. In certain embodiments, said method comprises performing a method according to the first aspect to thereby identify a modulator. In certain embodiments, said cell resides in or derives from a tissue that expresses human FPRL2. In certain embodiments, said tissue expressing human FPRL2 is selected from the group consisting of sciatic nerve, anterior hippocampus, whole brain, hippocampus, substantia nigra, spleen, heart, lung, pancreas, bone, and ovary.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is according to the fourth aspect. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is cell death-protective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less Man 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μm.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

In certain embodiments, said contacting comprises oral administration of said modulator to said individual.

The invention also relates to a method of preventing or treating cell proliferative disorder in an individual in need of said prevention or treatment, comprising contacting a therapeutically effective amount of a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, with the receptor. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a thirty-first aspect, the invention features a pharmaceutical or physiologically acceptable composition comprising, consisting essentially of, or consisting of a modulator a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence. In certain embodiments, said modulator is identifiable by performing a method according to the first aspect. In certain embodiments, said modulator is identified by performing a method according to the first aspect.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is according to the fourth aspect. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist.

In certain embodiments, said composition is pharmaceutical. In certain embodiments, said composition is physiologically acceptable.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is cell death-protective.

In certain embodiments, said modulator is neuroprotective.

In certain embodiments, said modulator is myoprotective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

In a thirty-second aspect, the invention features a method of reducing neuronal cell death comprising providing or administering to an individual in need of said reduction said pharmaceutical or physiologically acceptable composition of the thirty-first aspect.

The invention also features a method of preventing or treating a neuronal cell death-related disorder comprising providing or administering to an individual in need of said prevention or treatment said pharmaceutical or physiologically acceptable composition of the thirty-first aspect. In certain embodiments, said neuronal cell death-related disorder is selected from the group consisting of:

(a) Alzheimer's disease;

(b) Parkinson's disease;

(c) stroke;

(d) motor-neuron disease;

(e) learning or memory impairment;

(f) traumatic brain injury;

(g) spinal cord injury;

(h) peripheral neuropathy; and

(i) prion-associated disease.

In some embodiments, Alzheimer's disease encompasses mild cognitive impairment (MCI).

Motor-neuron disease includes, but is not limited to, amyotrophic lateral sclerosis, progressive spinal muscular atrophy, progressive bulbar palsy, and primary lateral sclerosis. Peripheral neuropathy includes, but is not limited, to diabetic neuropathy and neuropathy involving the sciatic nerve. Diabetic neuropathy includes, but is not limited to, distal symmetrical polyneuropathy (DSP).

In some embodiments, the neuronal cell death-related disorder is Alzheimer's disease. In some embodiments, the neuronal cell death-related disorder is Parkinson's disease. In some embodiments, the neuronal cell death-related disorder is stroke. In some embodiments, the neuronal cell death-related disorder is motor-neuron disease. In some embodiments, the neuronal cell death-related disorder is learning or memory impairment. In some embodiments, the neuronal cell death-related disorder is traumatic brain injury. In some embodiments, the neuronal cell death-related disorder is spinal cord injury. In some embodiments, the neuronal cell death-related disorder is peripheral neuropathy. In some embodiments, the neuronal cell death-related disorder is prion-associated disease.

In certain embodiments, said modulator is an agonist.

In certain embodiments, a therapeutically effective amount of said pharmaceutical or physiologically acceptable composition is provided or administered to said individual.

In certain embodiments, said providing or administering of said pharmaceutical or physiologically acceptable composition is oral.

The invention also relates to a method of preventing or treating a neuronal cell proliferative disorder comprising providing or administering to an individual in need of said prevention or treatment said pharmaceutical or physiologically acceptable composition of the thirty-first aspect. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist. In certain embodiments, said neuronal cell proliferative disorder is neuroblastoma. In other embodiments, said neuronal cell proliferative disorder is medulloblastoma.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a thirty-third aspect, the invention features a method of reducing muscle cell death comprising providing or administering to an individual in need of said reduction said pharmaceutical or physiologically acceptable composition of the thirty-first aspect.

The invention also features a method of preventing or treating a muscle cell death-related disorder comprising providing or administering to an individual in need of said prevention or treatment said pharmaceutical or physiologically acceptable composition of the thirty-first aspect. In certain embodiments, said cell death-related disorder is selected from the group consisting of:

(a) cerebral amyloid beta-protein angiopathy;

(b) myocardial infarction; and

(c) congestive heart failure.

In some embodiments, the muscle cell death-related disorder is cerebral amyloid beta-protein angiopathy. In some embodiments, the muscle cell death-related disorder is myocardial infarction. In some embodiments, the muscle cell death-related disorder is congestive heart failure.

In certain embodiments, said modulator is an agonist.

In certain embodiments, a therapeutically effective amount of said pharmaceutical or physiologically acceptable composition is provided or administered to said individual.

In certain embodiments, said providing or administering of said pharmaceutical or physiologically acceptable composition is oral.

The invention also relates to a method of preventing or treating a muscle cell proliferative disorder comprising providing or administering to an individual in need of said prevention or treatment said pharmaceutical or physiologically acceptable composition of the thirty first aspect. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist. In certain embodiments, said muscle cell proliferative disorder selected from the group consisting of:

(a) atherosclerosis;

(b) restenosis; and

(c) tumor-supportive angiogenesis.

In some embodiments, the muscle cell proliferative disorder is atherosclerosis. In some embodiments, the muscle cell proliferative disorder is restenosis. In some embodiments, the muscle cell proliferative disorder is tumor-supportive angiogenesis. In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a thirty-fourth aspect, the invention features a method of reducing cell death comprising providing or administering to an individual in need of said reduction said pharmaceutical or physiologically acceptable composition of the thirty-first aspect.

The invention also features a method of preventing or treating a cell death-related disorder comprising providing or administering to an individual in need of said prevention or treatment said pharmaceutical or physiologically acceptable composition of the thirty-first aspect. In certain, embodiments, said cell resides in or derives from a tissue that expresses human FPRL2. In certain embodiments, said tissue expressing human FPRL2 is selected from the group consisting of sciatic nerve, anterior hippocampus, whole brain, hippocampus, substantia nigra, spleen, heart, lung, pancreas, bone, and ovary.

In certain embodiments, a therapeutically effective amount of said pharmaceutical or physiologically acceptable composition is provided or administered to said individual.

In certain embodiments, said providing or administering of said pharmaceutical or physiologically acceptable composition is oral.

The invention also relates to a method of preventing or treating a cell proliferative disorder comprising providing or administering to an individual in need of said prevention or treatment said pharmaceutical or physiologically acceptable composition of the thirty-first aspect. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In an thirty-fifth aspect, the invention features a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for use in a method of treatment of the human or animal body by therapy. In certain embodiments, said modulator is identifiable by performing a method according to the first aspect. In certain embodiments, said modulator is identified by performing a method according to the first aspect.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is according to the fourth aspect. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

In certain embodiments, said animal is a mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, or non-human primate. More preferred of human or animal is human.

In an thirty-sixty aspect, the invention features a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for use in a method of reducing neuronal cell death in the human or animal body by therapy. In certain embodiments, said modulator is identifiable by performing a method according to the first aspect. In certain embodiments, said modulator is identified by performing a method according to the first aspect.

The invention also features a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for use in a method of prevention of or treatment for a neuronal cell death-related disorder in a human or animal body by therapy. In certain embodiments, said modulator is identifiable by performing a method according to the first aspect. In certain embodiments, said modulator is identified by performing a method according to the first aspect. In certain embodiments, said neuronal cell death-related disorder is selected from the group consisting of:

(a) Alzheimer's disease;

(b) Parkinson's disease;

(c) stroke;

(d) motor-neuron disease;

(e) learning or memory impairment;

(f) traumatic brain injury;

(g) spinal cord injury;

(h) peripheral neuropathy; and

(i) prion-associated disease.

In some embodiments, Alzheimer's disease encompasses mild cognitive impairment (MCI).

Motor-neuron disease includes, but is not limited to, amyotrophic lateral sclerosis, progressive spinal muscular atrophy, progressive bulbar palsy, and primary lateral sclerosis. Peripheral neuropathy includes, but is not limited, to diabetic neuropathy and neuropathy involving the sciatic nerve. Diabetic neuropathy includes, but is not limited to, distal symmetrical polyneuropathy (DSP).

In some embodiments, the neuronal cell death-related disorder is Alzheimer's disease. In some embodiments, the neuronal cell death-related disorder is Parkinson's disease. In some embodiments, the neuronal cell death-related disorder is stroke. In some embodiments, the neuronal cell death-related disorder is motor-neuron disease. In some embodiments, the neuronal cell death-related disorder is learning or memory impairment. In some embodiments, the neuronal cell death-related disorder is traumatic brain injury. In some embodiments, the neuronal cell death-related disorder is spinal cord injury. In some embodiments, the neuronal cell death-related disorder is peripheral neuropathy. In some embodiments, the neuronal cell death-related disorder is prion-associated disease.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is according to the fourth aspect. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is neuroprotective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ HD NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC 50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

The invention also relates to a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for use in a method of prevention of or treatment for a neuronal cell proliferative disorder in a human or animal body by therapy. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist. In certain embodiments, said neuronal cell proliferative disorder is neuroblastoma. In other embodiments, said neuronal cell proliferative disorder is medulloblastoma.

In certain embodiments, said animal is a mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, or non-human primate. More preferred of human or animal is human.

In an thirty-seventh aspect, the invention features a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for use in a method of reducing muscle cell death in the human or animal body by therapy. In certain embodiments, said modulator is identifiable by performing a method according to the first aspect. In certain embodiments, said modulator is identified by performing a method according to the first aspect.

The invention also features a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for use in a method of prevention of or treatment for a muscle cell death-related disorder in a human or animal body by therapy. In certain embodiments, said modulator is identifiable by performing a method according to the first aspect. In certain embodiments, said modulator is identified by performing a method according to the first aspect. In certain embodiments, said muscle cell death-related disorder is selected from the group consisting of:

(a) cerebral amyloid beta-protein angiopathy;

(b) myocardial infarction; and

(c) congestive heart failure.

In some embodiments, the muscle cell death-related disorder is cerebral amyloid beta-protein angiopathy. In some embodiments, the muscle cell death-related disorder is myocardial infarction. In some embodiments, the muscle cell death-related disorder is congestive heart failure.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is according to the fourth aspect. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is myoprotective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

The invention also relates to a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for use in a method of prevention of or treatment for a muscle cell proliferative disorder in a human or animal body by therapy. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist. In certain embodiments, said muscle cell proliferative disorder selected from the group consisting of:

(a) atherosclerosis;

(b) restenosis; and

(c) tumor-supportive angiogenesis.

In some embodiments, the muscle cell proliferative disorder is atherosclerosis. In some embodiments, the muscle cell proliferative disorder is restenosis. In some embodiments, the muscle cell proliferative disorder is tumor-supportive angiogenesis.

In certain embodiments, said animal is a mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, or non-human primate. More preferred of human or animal is human.

In a thirty-eighth aspect, the invention features a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for use in a method of reducing cell death in the human or animal body by therapy. In certain embodiments, said modulator is identifiable by performing a method according to the first aspect. In certain embodiments, said modulator is identified by performing a method according to the first aspect.

The invention also features a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for use in a method of prevention of or treatment for a cell death-related disorder in a human or animal body by therapy. In certain embodiments, said modulator is identifiable by performing a method according to the first aspect. In certain embodiments, said modulator is identified by performing a method according to the first aspect. In certain embodiments, said cell resides in or derives from a tissue that expresses human FPRL2. In certain embodiments, said tissue expressing human FPRL2 is selected from the group consisting of sciatic nerve, anterior hippocampus, whole brain, hippocampus, substantia nigra, spleen, heart, lung, pancreas, bone, and ovary.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is according to the fourth aspect. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 50%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is cell death-protective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 M in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

The invention also relates to a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for use in a method of prevention of or treatment for a cell proliferative disorder in a human or animal body by therapy. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist.

In certain embodiments, said animal is a mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, or non-human primate. More preferred of human or animal is human.

In a thirty-ninth aspect, the invention features a method of using a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for the preparation of a medicament for the reduction of neuronal cell death. In certain embodiments, said method comprises performing a method according to the first aspect to thereby identify a modulator.

The invention also features a method of using a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for the preparation of a medicament for the prevention or treatment of a neuronal cell death-related disorder. In certain embodiments, said method comprises performing a method according to the first aspect to thereby identify a modulator. In certain embodiments, said neuronal cell death-related disorder is selected from the group consisting of:

(a) Alzheimer's disease; (b) Parkinson's disease; (c) stroke; (d) motor-neuron disease; (e) learning or memory impairment; (d) traumatic brain injury; (e) spinal cord injury; (f) peripheral neuropathy; and (g) prion-associated disease.

In some embodiments, the neuronal cell death-related disorder is Alzheimer's disease. In some embodiments, the neuronal cell death-related disorder is Parkinson's disease. In some embodiments, the neuronal cell death-related disorder is stroke. In some embodiments, the neuronal cell death-related disorder is motor-neuron disease. In some embodiments, the neuronal cell death-related disorder is learning or memory impairment. In some embodiments, the neuronal cell death-related disorder is traumatic brain injury. In some embodiments, the neuronal cell death-related disorder is spinal cord injury. In some embodiments, the neuronal cell death-related disorder is peripheral neuropathy. In some embodiments, the neuronal cell death-related disorder is prion-associated disease.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is according to the fourth aspect. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is neuroprotective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

In some embodiments, Alzheimer's disease encompasses mild cognitive impairment (MCI).

Motor-neuron disease includes, but is not limited to, amyotrophic lateral sclerosis, progressive spinal muscular atrophy, progressive bulbar palsy, and primary lateral sclerosis. Peripheral neuropathy includes, but is not limited, to diabetic neuropathy and neuropathy involving the sciatic nerve. Diabetic neuropathy includes, but is not limited to, distal symmetrical polyneuropathy (DSP).

The invention also features a method of using a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for the preparation of a medicament for preventing or treating a neuronal cell proliferative disorder. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist. In certain embodiments, said neuronal cell proliferative disorder is neuroblastoma. In other embodiments, said neuronal cell proliferative disorder is medulloblastoma.

In a fortieth aspect, the invention features a method of using a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for the preparation of a medicament for the reduction of muscle cell death. In certain embodiments, said method comprises performing a method according to the first aspect to thereby identify a modulator.

The invention also features a method of using a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for the preparation of a medicament for the prevention or treatment of a muscle cell death-related disorder. In certain embodiments, said method comprises performing a method according to the first aspect to thereby identify a modulator. In certain embodiments, said muscle cell death-related disorder is selected from the group consisting of:

(a) cerebral amyloid beta-protein angiopathy; (b) myocardial infarction; and (c) congestive heart failure.

In some embodiments, the muscle cell death-related disorder is cerebral amyloid beta-protein angiopathy. In some embodiments, the muscle cell death-related disorder is myocardial infarction. In some embodiments, the muscle cell death-related disorder is congestive heart failure.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is according to the fourth aspect. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is myoprotective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

The invention also relates to a method of using a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for the preparation of a medicament for the prevention or treatment of a muscle cell proliferative disorder. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist. In certain embodiments, said muscle cell proliferative disorder is selected from the group consisting of:

(a) atherosclerosis;

(b) restenosis; and

(c) tumor-supportive angiogenesis.

In some embodiments, the muscle cell proliferative disorder is atherosclerosis. In some embodiments, the muscle cell proliferative disorder is restenosis. In some embodiments, the muscle cell proliferative disorder is tumor-supportive angiogenesis.

In a forty-first aspect, the invention features a method of using a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for the preparation of a medicament for the reduction of cell death. In certain embodiments, said method comprises performing a method according to the first aspect to thereby identify a modulator.

The invention also features a method of using a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for the preparation of a medicament for the prevention or treatment of a cell death-related disorder. In certain embodiments, said method comprises performing a method according to the first aspect to thereby identify a modulator. In certain embodiments, said cell resides in or derives from a tissue that expresses human FPRL2. In certain embodiments, said tissue expressing human FPRL2 is selected from the group consisting of sciatic nerve, anterior hippocampus, whole brain, hippocampus, substantia nigra, spleen, heart, lung, pancreas, bone, and ovary.

In certain embodiments, said modulator is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof. In certain embodiments, said modulator is not a peptide. In certain embodiments, said modulator is according to the fourth aspect. In certain embodiments, said modulator is selected from the group consisting of agonist, partial agonist, inverse agonist, and antagonist. In certain preferred embodiments, said modulator is an agonist.

In certain embodiments, said modulator is selective for the GPCR.

In certain embodiments, said modulator is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable modulator is further able to cross the blood-brain barrier.

In certain embodiments, said modulator is cell death-protective.

In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said EC50 is in an assay selected from the group consisting of: GTPγS binding assay carried out with membrane from transfected CHO cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; melanophore assay carried out using transfected melanophores expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6; and whole cell cAMP assay carried using transfected HEK293 cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:2 or 6. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM, of less than 10 μM, or of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 100 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 90 μM in said assay. In some embodiments, said modulator is an inverse agonist or antagonist with an EC50 of less than 80 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 70 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 60 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 50 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 40 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 30 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 20 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 10 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 9 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 8 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 7 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 6 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 5 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 4 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 3 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 2 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 of less than 1 μM in said assay. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 100 μM. In some embodiments, said modulator is an agonist with an EC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM.

In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 of less than a value selected from the interval of 1 μM to 100 μM in an assay selected from the group consisting of GTPγS binding assay, melanophore assay, and whole cell cAMP assay, wherein said assay is carried out using transfected cells expressing recombinant Humanin GPCR polypeptide having the amino acid sequence of SEQ ID NO:12. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 100 μM, less than 90 μM, less than 80 μM, less than 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than 30 μM, less than 20 μM, or less than 10 μM. In some embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than a value selected from the interval of 1 μM to 10 μM. In certain embodiments, said modulator is an inverse agonist or an antagonist with an IC 50 in said assay of less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, or less than 1 μM.

The invention also relates to a method of using a modulator of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, for the preparation of a medicament for the prevention or treatment of a cell proliferative disorder. In certain embodiments, said modulator is an inverse agonist. In certain embodiments, said modulator is an antagonist.

In a forty-second aspect, the invention features a method of preparing a pharmaceutical or physiologically acceptable composition comprising admixing a compound according to the second aspect and a carrier.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In a forty-third aspect, the invention features a method of preparing a pharmaceutical or physiologically acceptable composition comprising admixing a compound according the third aspect and a carrier.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In a forty-fourths aspect, the invention features a pharmaceutical or physiologically acceptable composition comprising, consisting essentially of, or consisting of a compound according to the second aspect.

In certain embodiments, said composition is pharmaceutical. In certain embodiments, said composition is physiologically acceptable.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In a forty-fifth aspect, the invention features a pharmaceutical or physiologically acceptable composition comprising, consisting essentially of, or consisting of a compound according to the third aspect.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In a forty-sixth aspect, the invention features a method of modulating the activity of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for reducing neuronal cell death in an individual in need of said modulation, comprising contacting said receptor with a therapeutically effective amount of a compound according to the second or third aspect or with a therapeutically effect amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth or forty-fifth aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the second aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the third aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fifth aspect.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a forty-seventh aspect, the invention features a method of modulating the activity of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for preventing or treating a neuronal cell death-related disorder in an individual in need of said modulation, comprising contacting said receptor with a therapeutically effective amount of a compound according to the second or third aspect or with a therapeutically effect amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth or forty-fifth aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the second aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the third aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fifth aspect. In certain embodiments, said neuronal cell death-related disorder is selected from the group consisting of:

(a) Alzheimer's disease;

(b) Parkinson's disease;

(c) stroke;

(d) motor-neuron disease;

(e) learning or memory impairment;

(f) traumatic brain injury;

(g) spinal cord injury;

(h) peripheral neuropathy; and

(i) prion-associated disease.

In some embodiments, Alzheimer's disease encompasses mild cognitive impairment (MCI).

Motor-neuron disease includes, but is not limited to, amyotrophic lateral sclerosis, progressive spinal muscular atrophy, progressive bulbar palsy, and primary lateral sclerosis. Peripheral neuropathy includes, but is not limited, to diabetic neuropathy and neuropathy involving the sciatic nerve. Diabetic neuropathy includes, but is not limited to, distal symmetrical polyneuropathy (DSP).

In some embodiments, the neuronal cell death-related disorder is Alzheimer's disease. In some embodiments, the neuronal cell death-related disorder is Parkinson's disease. In some embodiments, the neuronal cell death-related disorder is stroke. In some embodiments, the neuronal cell death-related disorder is motor-neuron disease. In some embodiments, the neuronal cell death-related disorder is learning or memory impairment. In some embodiments, the neuronal cell death-related disorder is traumatic brain injury. In some embodiments, the neuronal cell death-related disorder is spinal cord injury. In some embodiments, the neuronal cell death-related disorder is peripheral neuropathy. In some embodiments, the neuronal cell death-related disorder is prion-associated disease.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a forty-eighth aspect, the invention features a method of modulating the activity of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for reducing muscle cell death in an individual in need of said modulation, comprising contacting said receptor with a therapeutically effective amount of a compound according to the second or third aspect or with a therapeutically effect amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth or forty-fifth aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the second aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the third aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fifth aspect.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a forty-ninth aspect, the invention features a method of modulating the activity of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for preventing or treating a muscle cell death-related disorder in an individual in need of said modulation, comprising contacting said receptor with a therapeutically effective amount of a compound according to the second or third aspect or with a therapeutically effect amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth or forty-fifth aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the second aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the third aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fifth aspect. In certain embodiments, said muscle cell death-related disorder is selected from the group consisting of:

(a) cerebral amyloid beta-protein angiopathy; (b) myocardial infarction; and (c) congestive heart failure.

In some embodiments, the muscle cell death-related disorder is cerebral amyloid beta-protein angiopathy. In some embodiments, the muscle cell death-related disorder is myocardial infarction. In some embodiments, the muscle cell death-related disorder is congestive heart failure.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a fiftieth aspect, the invention features a method of modulating the activity of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for reducing cell death in an individual in need of said modulation, comprising contacting said receptor with a therapeutically effective amount of a compound according to the second or third aspect or with a therapeutically effect amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth or forty-fifth aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the second aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the third aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fifth aspect. In certain embodiments, said cell resides in or derives from a tissue that expresses human FPRL2. In certain embodiments, said tissue expressing human FPRL2 is selected from the group consisting of sciatic nerve, anterior hippocampus, whole brain, hippocampus, substantia nigra, spleen, heart, lung, pancreas, bone, and ovary.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a fifty-first aspect, the invention features a method of modulating the activity of a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, wherein said modulation is for preventing or treating a cell death-related disorder in an individual in need of said modulation, comprising contacting said receptor with a therapeutically effective amount of a compound according to the second or third aspect or with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth or forty-fifth aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the second aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the third aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fifth aspect. In certain embodiments, said cell resides in or derives from a tissue that expresses human FPRL2. In certain embodiments, said tissue expressing human FPRL2 is selected from the group consisting of sciatic nerve, anterior hippocampus, whole brain, hippocampus, substantia nigra, spleen, heart, lung, pancreas, bone, and ovary.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 10%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a fifty-second aspect, the invention features a method of reducing neuronal cell death in an individual in need of said reducing, comprising contacting said receptor with a therapeutically effective amount of a compound according to the second or third aspect or with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth or forty-fifth aspect with a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the second aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the third aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fifth aspect.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a fifth-third aspect, the invention features a method of preventing or treating a neuronal cell death-related disorder in an individual in need of said reducing, comprising contacting said receptor with a therapeutically effective amount of a compound according to the second or third aspect or with a therapeutically effect amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth or forty-fifth aspect with a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the second aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the third aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fifth aspect. In certain embodiments, said neuronal cell death-related disorder is selected from the group consisting of:

(a) Alzheimer's disease;

(b) Parkinson's disease;

(c) stroke;

(d) motor-neuron disease;

(e) learning or memory impairment;

(f) traumatic brain injury;

(g) spinal cord injury;

(h) peripheral neuropathy; and

(i) prion-associated disease.

In some embodiments, Alzheimer's disease encompasses mild cognitive impairment (MCI).

Motor-neuron disease includes, but is not limited to, amyotrophic lateral sclerosis, progressive spinal muscular atrophy, progressive bulbar palsy, and primary lateral sclerosis. Peripheral neuropathy includes, but is not limited, to diabetic neuropathy and neuropathy involving the sciatic nerve. Diabetic neuropathy includes, but is not limited to, distal symmetrical polyneuropathy (DSP).

In some embodiments, the neuronal cell death-related disorder is Alzheimer's disease. In some embodiments, the neuronal cell death-related disorder is Parkinson's disease. In some embodiments, the neuronal cell death-related disorder is stroke. In some embodiments, the neuronal cell death-related disorder is motor-neuron disease. In some embodiments, the neuronal cell death-related disorder is learning or memory impairment. In some embodiments, the neuronal cell death-related disorder is traumatic brain injury. In some embodiments, the neuronal cell death-related disorder is spinal cord injury. In some embodiments, the neuronal cell death-related disorder is peripheral neuropathy. In some embodiments, the neuronal cell death-related disorder is prion-associated disease.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a fifty-fourth aspect, the invention features a method of reducing muscle cell death in an individual in need of said reducing, comprising contacting said receptor with a therapeutically effective amount of a compound according to the second or third aspect or with a therapeutically effect amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth or forty-fifth aspect with a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the second aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the third aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fifth aspect.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a fifty-fifth aspect, the invention features a method of preventing or treating a muscle cell death-related disorder in an individual in need of said reducing, comprising contacting said receptor with a therapeutically effective amount of a compound according to the second or third aspect or with a therapeutically effect amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth or forty-fifth aspect with a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the second aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the third aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fifth aspect. In certain embodiments, said muscle cell death-related disorder is selected from the group consisting of:

(a) cerebral amyloid beta-protein angiopathy; (b) myocardial infarction; and (c) congestive heart failure.

In some embodiments, the muscle cell death-related disorder is cerebral amyloid beta-protein angiopathy. In some embodiments, the muscle cell death-related disorder is myocardial infarction. In some embodiments, the muscle cell death-related disorder is congestive heart failure.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a fifty-sixth aspect, the invention features a method of reducing cell death in an individual in need of said reducing, comprising contacting said receptor with a therapeutically effective amount of a compound according to the second or third aspect or with a therapeutically effect amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth or forty-fifth aspect with a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the second aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the third aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fifth aspect. In certain embodiments, said cell resides in or derives from a tissue that expresses human FPRL2. In certain embodiments, said tissue expressing human FPRL2 is selected from the group consisting of sciatic nerve, anterior hippocampus, whole brain, hippocampus, substantia nigra, spleen, heart, lung, pancreas, bone, and ovary.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human-mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a fifty-seventh aspect, the invention features a method of preventing or treating a cell death-related disorder in an individual in need of said reducing, comprising contacting said receptor with a therapeutically effective amount of a compound according to the second or third aspect or with a therapeutically effect amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth or forty-fifth aspect with a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the second aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a compound according to the third aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth aspect. In certain embodiments, said contacting is with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fifth aspect. In certain embodiments, said cell resides in or derives from a tissue that expresses human FPRL2. In certain embodiments, said tissue expressing human FPRL2 is selected from the group consisting of sciatic nerve, anterior hippocampus, whole brain, hippocampus, substantia nigra, spleen, heart, lung, pancreas, bone, and ovary.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a fifty-eighth aspect, the invention features a method of reducing neuronal cell death comprising providing or administering to an individual in need of said reducing a compound according to the second or third aspect or with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth or forty-fifth aspect. In certain embodiments, said providing or administering a compound is providing or administering a compound according to the second aspect. In certain embodiments, said providing or administering a compound is providing or administering a compound according to the third aspect. In certain embodiments, said providing or administering a pharmaceutical or physiologically acceptable composition is providing or administering a pharmaceutical or physiologically acceptable composition according to the forty-fourth aspect. In certain embodiments, said providing or administering a pharmaceutical or physiologically acceptable composition is providing or administering a pharmaceutical or physiologically acceptable composition according to the forty-fifth aspect.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a fifty-ninth aspect, the invention features a method of treating a neuronal cell death-related disorder comprising providing or administering to an individual in need of said treating or preventing a compound according to the second or third aspect or with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth or forty-fifth aspect. In certain embodiments, said providing or administering a compound is providing or administering a compound according to the second aspect. In certain embodiments, said providing or administering a compound is providing or administering a compound according to the third aspect. In certain embodiments, said providing or administering a pharmaceutical or physiologically acceptable composition is providing or administering a pharmaceutical or physiologically acceptable composition according to the forty-fourth aspect. In certain embodiments, said providing or administering a pharmaceutical or physiologically acceptable composition is providing or administering a pharmaceutical or physiologically acceptable composition according to the forty-fifth aspect. In certain embodiments, said neuronal cell death-related disorder is selected from the group consisting of:

(a) Alzheimer's disease;

(b) Parkinson's disease;

(c) stroke;

(d) motor-neuron disease;

(e) learning or memory impairment;

(f) traumatic brain injury;

(g) spinal cord injury;

(h) peripheral neuropathy; and

(i) prion-associated disease.

In some embodiments, Alzheimer's disease encompasses mild cognitive impairment (MCI).

Motor-neuron disease includes, but is not limited to, amyotrophic lateral sclerosis, progressive spinal muscular atrophy, progressive bulbar palsy, and primary lateral sclerosis. Peripheral neuropathy includes, but is not limited, to diabetic neuropathy and neuropathy involving the sciatic nerve. Diabetic neuropathy includes, but is not limited to, distal symmetrical polyneuropathy (DSP).

In some embodiments, the neuronal cell death-related disorder is Alzheimer's disease. In some embodiments, the neuronal cell death-related disorder is Parkinson's disease. In some embodiments, the neuronal cell death-related disorder is stroke. In some embodiments, the neuronal cell death-related disorder is motor-neuron disease. In some embodiments, the neuronal cell death-related disorder is learning or memory impairment. In some embodiments, the neuronal cell death-related disorder is traumatic brain injury. In some embodiments, the neuronal cell death-related disorder is spinal cord injury. In some embodiments, the neuronal cell death-related disorder is peripheral neuropathy. In some embodiments, the neuronal cell death-related disorder is prion-associated disease.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a sixtieth aspect, the invention features a method of reducing muscle cell death comprising providing or administering to an individual in need of said reducing a compound according to the second or third aspect or with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth or forty-fifth aspect. In certain embodiments, said providing or administering a compound is providing or administering a compound according to the second aspect. In certain embodiments, said providing or administering a compound is providing or administering a compound according to the third aspect. In certain embodiments, said providing or administering a pharmaceutical or physiologically acceptable composition is providing or administering a pharmaceutical or physiologically acceptable composition according to the forty-fourth aspect. In certain embodiments, said providing or administering a pharmaceutical or physiologically acceptable composition is providing or administering a pharmaceutical or physiologically acceptable composition according to the forty-fifth aspect.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a sixty-first aspect, the invention features a method of treating a muscle cell death-related disorder comprising providing or administering to an individual in need of said treating or preventing a compound according to the second or third aspect or with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth or forty-fifth aspect. In certain embodiments, said providing or administering a compound is providing or administering a compound according to the second aspect. In certain embodiments, said providing or administering a compound is providing or administering a compound according to the third aspect. In certain embodiments, said providing or administering a pharmaceutical or physiologically acceptable composition is providing or administering a pharmaceutical or physiologically acceptable composition according to the forty-fourth aspect. In certain embodiments, said providing or administering a pharmaceutical or physiologically acceptable composition is providing or administering a pharmaceutical or physiologically acceptable composition according to the forty-fifth aspect. In certain embodiments, said muscle cell death-related disorder is selected from the group consisting of:

(a) cerebral amyloid beta-protein angiopathy; (b) myocardial infarction; and (c) congestive heart failure.

In some embodiments, the muscle cell death-related disorder is cerebral amyloid beta-protein angiopathy. In some embodiments, the muscle cell death-related disorder is myocardial infarction. In some embodiments, the muscle cell death-related disorder is congestive heart failure.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a sixty-second aspect, the invention features a method of reducing cell death comprising providing or administering to an individual in need of said reducing a compound according to the second or third aspect or with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth or forty-fifth aspect. In certain embodiments, said providing or administering a compound is providing or administering a compound according to the second aspect. In certain embodiments, said providing or administering a compound is providing or administering a compound according to the third aspect. In certain embodiments, said providing or administering a pharmaceutical or physiologically acceptable composition is providing or administering a pharmaceutical or physiologically acceptable composition according to the forty-fourth aspect. In certain embodiments, said providing or administering a pharmaceutical or physiologically acceptable composition is providing or administering a pharmaceutical or physiologically acceptable composition according to the forty-fifth aspect. In certain embodiments, said cell resides in or derives from a tissue that expresses human FPRL2. In certain embodiments, said tissue expressing human FPRL2 is selected from the group consisting of sciatic nerve, anterior hippocampus, whole brain, hippocampus, substantia nigra, spleen, heart, lung, pancreas, bone, and ovary.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a sixty-third aspect, the invention features a method of treating a cell death-related disorder comprising providing or administering to an individual in need of said treating or preventing a compound according to the second or third aspect or with a therapeutically effective amount of a pharmaceutical or physiologically acceptable composition according to the forty-fourth or forty-fifth aspect. In certain embodiments, said providing or administering a compound is providing or administering a compound according to the second aspect. In certain embodiments, said providing or administering a compound is providing or administering a compound according to the third aspect. In certain embodiments, said providing or administering a pharmaceutical or physiologically acceptable composition is providing or administering a pharmaceutical or physiologically acceptable composition according to the forty-fourth aspect. In certain embodiments, said providing or administering a pharmaceutical or physiologically acceptable composition is providing or administering a pharmaceutical or physiologically acceptable composition according to the forty-fifth aspect. In certain embodiments, said cell resides in or derives from a tissue that expresses human FPRL2. In certain embodiments, said tissue expressing human FPRL2 is selected from the group consisting of sciatic nerve, anterior hippocampus, whole brain, hippocampus, substantia nigra, spleen, heart, lung, pancreas, bone, and ovary.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said individual is a mammal. In certain embodiments, said individual is a non-human mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, non-human primate or human. In certain embodiments, said mammal is a mouse, rat, non-human primate, or human. Most preferred is human.

In a sixty-fourth aspect, the invention features a compound according to the second or third aspect for use in a method of treatment of the human or animal body by therapy. In certain embodiments, said compound is according to the second aspect. In certain embodiments, said compound is according to the third aspect.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said animal is a mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, or non-human primate. More preferred of human or animal is human.

In a sixty-fifth aspect, the invention features a compound according to the second or third aspect for use in a method of reducing neuronal cell death in the human or animal body by therapy. In certain embodiments, said compound is according to the second aspect. In certain embodiments, said compound is according to the third aspect.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said animal is a mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, or non-human primate. More preferred of human or animal is human.

In a sixty-sixth aspect, the invention features a compound according to the second or third aspect for use in a method of prevention or treatment for a neuronal cell death-related disorder in the human or animal body by therapy. In certain embodiments, said compound is according to the second aspect. In certain embodiments, said compound is according to the third aspect. In certain embodiments, said neuronal cell death-related disorder is selected from the group consisting of:

(a) Alzheimer's disease;

(b) Parkinson's disease;

(c) stroke;

(d) motor-neuron disease;

(e) learning or memory impairment;

(f) traumatic brain injury;

(g) spinal cord injury;

(h) peripheral neuropathy; and

(i) prion-associated disease.

In some embodiments, Alzheimer's disease encompasses mild cognitive impairment (MCI).

Motor-neuron disease includes, but is not limited to, amyotrophic lateral sclerosis, progressive spinal muscular atrophy, progressive bulbar palsy, and primary lateral sclerosis. Peripheral neuropathy includes, but is not limited, to diabetic neuropathy and neuropathy involving the sciatic nerve. Diabetic neuropathy includes, but is not limited to, distal symmetrical polyneuropathy (DSP).

In some embodiments, the neuronal cell death-related disorder is Alzheimer's disease. In some embodiments, the neuronal cell death-related disorder is Parkinson's disease. In some embodiments, the neuronal cell death-related disorder is stroke. In some embodiments, the neuronal cell death-related disorder is motor-neuron disease. In some embodiments, the neuronal cell death-related disorder is learning or memory impairment. In some embodiments, the neuronal cell death-related disorder is traumatic brain injury. In some embodiments, the neuronal cell death-related disorder is spinal cord injury. In some embodiments, the neuronal cell death-related disorder is peripheral neuropathy. In some embodiments, the neuronal cell death-related disorder is prion-associated disease.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said animal is a mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, or non-human primate. More preferred of human or animal is human.

In a sixty-seventh aspect, the invention features a compound according to the second or third aspect for use in a method of prevention or treatment of muscle cell death in the human or animal body by therapy. In certain embodiments, said compound is according to the second aspect. In certain embodiments, said compound is according to the third aspect.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said animal is a mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, or non-human primate. More preferred of human or animal is human.

In a sixty-eighth aspect, the invention features a compound according to the second or third aspect for use in a method of prevention or treatment for a muscle cell death-related disorder in the human or animal body by therapy. In certain embodiments, said compound is according to the second aspect. In certain embodiments, said compound is according to the third aspect. In certain embodiments, said muscle cell death-related disorder is selected from the group consisting of:

(a) cerebral amyloid beta-protein angiopathy; (b) myocardial infarction; and (c) congestive heart failure.

In some embodiments, the muscle cell death-related disorder is cerebral amyloid beta-protein angiopathy. In some embodiments, the muscle cell death-related disorder is myocardial infarction. In some embodiments, the muscle cell death-related disorder is congestive heart failure.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said animal is a mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, or non-human primate. More preferred of human or animal is human.

In a sixty-ninth aspect, the invention features a compound according to the second or third aspect for use in a method of prevention or treatment of cell death in the human or animal body by therapy. In certain embodiments, said compound is according to the second aspect. In certain embodiments, said compound is according to the third aspect. In certain embodiments, said cell resides in or derives from a tissue that expresses human FPRL2. In certain embodiments, said tissue expressing human FPRL2 is selected from the group consisting of sciatic nerve, anterior hippocampus, whole brain, hippocampus, substantia nigra, spleen, heart, lung, pancreas, bone, and ovary.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said animal is a mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, or non-human primate. More preferred of human or animal is human.

In a seventieth aspect, the invention features a compound according to the second or third aspect for use in a method of prevention or treatment for a cell death-related disorder in the human or animal body by therapy. In certain embodiments, said compound is according to the second aspect. In certain embodiments, said compound is according to the third aspect. In certain embodiments, said cell resides in or derives from a tissue that expresses human FPRL2. In certain embodiments, said tissue expressing human FPRL2 is selected from the group consisting of sciatic nerve, anterior hippocampus, whole brain, hippocampus, substantia nigra, spleen, heart, lung, pancreas, bone, and ovary.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In certain embodiments, said animal is a mammal. In certain embodiments, said mammal is a horse, cow, sheep, pig, cat, dog, rabbit, mouse, rat, or non-human primate. More preferred of human or animal is human.

In a seventy-first aspect, the invention features a method of using a compound according to the second or third aspect for the preparation of a medicament for the reduction of neuronal cell death. In certain embodiments, said compound is according to the second aspect. In certain embodiments, said compound is according to the third aspect.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In a seventy-second aspect, the invention features a method of using a compound according to the second or third aspect for the preparation of a medicament for the prevention of or treatment of a neuronal cell death-related disorder. In certain embodiments, said compound is according to the second aspect. In certain embodiments, said compound is according to the third aspect. In certain embodiments, said neuronal cell death-related disorder is selected from the group consisting of:

(a) Alzheimer's disease;

(b) Parkinson's disease;

(c) stroke;

(d) motor-neuron disease;

(e) learning or memory impairment;

(f) traumatic brain injury;

(g) spinal cord injury;

(h) peripheral neuropathy; and

(i) prion-associated disease.

In some embodiments, Alzheimer's disease encompasses mild cognitive impairment (MCI).

Motor-neuron disease includes, but is not limited to, amyotrophic lateral sclerosis, progressive spinal muscular atrophy, progressive bulbar palsy, and primary lateral sclerosis. Peripheral neuropathy includes, but is not limited, to diabetic neuropathy and neuropathy involving the sciatic nerve. Diabetic neuropathy includes, but is not limited to, distal symmetrical polyneuropathy (DSP).

In some embodiments, the neuronal cell death-related disorder is Alzheimer's disease. In some embodiments, the neuronal cell death-related disorder is Parkinson's disease. In some embodiments, the neuronal cell death-related disorder is stroke. In some embodiments, the neuronal cell death-related disorder is motor-neuron disease. In some embodiments, the neuronal cell death-related disorder is learning or memory impairment. In some embodiments, the neuronal cell death-related disorder is traumatic brain injury. In some embodiments, the neuronal cell death-related disorder is spinal cord injury. In some embodiments, the neuronal cell death-related disorder is peripheral neuropathy. In some embodiments, the neuronal cell death-related disorder is prion-associated disease.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In a seventy-third aspect, the invention features a method of using a compound according to the second or third aspect for the preparation of a medicament for the reduction of muscle cell death. In certain embodiments, said compound is according to the second aspect. In certain embodiments, said compound is according to the third aspect.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In a seventy-fourth aspect, the invention features a method of using a compound according to the second or third aspect for the preparation of a medicament for the prevention of or treatment of a muscle cell death-related disorder. In certain embodiments, said compound is according to the second aspect. In certain embodiments, said compound is according to the third aspect. In certain embodiments, said muscle cell death-related disorder is selected from the group consisting of:

(a) cerebral amyloid beta-protein angiopathy; (b) myocardial infarction; and (c) congestive heart failure.

In some embodiments, the muscle cell death-related disorder is cerebral amyloid beta-protein angiopathy. In some embodiments, the muscle cell death-related disorder is myocardial infarction. In some embodiments, the muscle cell death-related disorder is congestive heart failure.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In a seventy-fifth aspect, the invention features a method of using a compound according to the second or third aspect for the preparation of a medicament for the reduction of cell death. In certain embodiments, said compound is according to the second aspect. In certain embodiments, said compound is according to the third aspect. In certain embodiments, said cell resides in or derives from a tissue that expresses human FPRL2. In certain embodiments, said tissue expressing human FPRL2 is selected from the group consisting of sciatic nerve, anterior hippocampus, whole brain, hippocampus, substantia nigra, spleen, heart, lung, pancreas, bone, and ovary.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In a seventy-sixth aspect, the invention features a method of using a compound according to the second or third aspect for the preparation of a medicament for the prevention of or treatment of a cell death-related disorder. In certain embodiments, said compound is according to the second aspect. In certain embodiments, said compound is according to the third aspect. In certain embodiments, said cell resides in or derives from a tissue that expresses human FPRL2. In certain embodiments, said tissue expressing human FPRL2 is selected from the group consisting of sciatic nerve, anterior hippocampus, whole brain, hippocampus, substantia nigra, spleen, heart, lung, pancreas, bone, and ovary.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In a seventy-seventh aspect, the invention features a method of modulating a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, comprising contacting said receptor with a compound according to the second or third aspect or with a pharmaceutical or physiologically acceptable composition according to the forty-fourth or forty-fifth aspect. In certain embodiments, said contacting is with a compound according to the second aspect. In certain embodiments, said contacting is with a compound according to the third aspect. In certain embodiments, said contacting is with a pharmaceutical or physiologically acceptable composition according to the forty-fourth aspect. In certain embodiments, said contacting is with a pharmaceutical or physiologically acceptable composition according to the forty-fifth aspect.

In certain embodiments, said compound is orally bioavailable. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intraperitoneal administration. In some embodiments, said oral bioavailability is at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration. In some embodiments, said oral bioavailablity is at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to intravenous administration.

In certain embodiments, said orally bioavailable compound is further able to cross the blood-brain barrier.

In a seventy-eighth aspect, the invention features a method of identifying whether a candidate compound binds to a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, comprising the steps of:

(a) contacting the receptor with a detectably labeled known ligand of the GPCR in the presence or absence of the candidate compound; and (b) determining whether the binding of said labeled ligand is inhibited in the presence of the candidate compound;

wherein said inhibition is indicative of the candidate compound binding to a Humanin GPCR.

In certain embodiments, the Humanin GPCR is recombinant. In certain embodiments, said contacting comprises contacting with a host cell or with membrane of a host cell that expresses the GPCR, wherein said host cell comprises an expression vector comprising a polynucleotide encoding the receptor.

The invention also relates to a method of determining whether a candidate compound binds to a Humanin GPCR, comprising the steps of:

(a) culturing Humanin GPCR-expressing host cells under conditions that would allow expression of a recombinant Humanin GPCR, said host cells being transfected with an expression vector comprising a polynucleotide encoding said recombinant Humanin GPCR comprising an FPRL2 amino acid sequence; (b) exposing a first population of Humanin GPCR-expressing cells of step (a) to a detectably labeled known ligand of said Humanin GPCR; (c) exposing a second population of Humanin GPCR-expressing cells of step (a) to the compound and the labeled known ligand of said Humanin GPCR of step (b); (d) determining the binding of the labeled known ligand of said Humanin GPCR-expressing cells of step (b) and step (c); and (e) comparing the binding of the labeled known ligand to said Humanin GPCR to the Humanin GPCR-expressing cells of step (b) and step (c);

wherein inhibition of binding of the labeled known ligand of said Humanin GPCR in the presence of the compound is indicative of the compound binding to a Humanin GPCR.

The invention also relates to a method of determining whether a candidate compound binds to a Humanin GPCR, comprising the steps of:

(a) culturing Humanin GPCR-expressing host cells under conditions that would allow expression of a recombinant Humanin GPCR, said host cells being transfected with an expression vector comprising a polynucleotide encoding said recombinant Humanin GPCR comprising an FPRL2 amino acid sequence; (b) preparing membrane from the Humanin GPCR-expressing cells of step (a); (c) exposing a first population of the membrane preparation of step (b) to a detectably labeled known ligand of said Humanin GPCR of step (b); (d) exposing a second population of the membrane preparation of step (b) to the candidate compound and the labeled known ligand of said Humanin GPCR; (e) determining the binding of the labeled known ligand of said Humanin GPCR to the membrane preparations of step (c) and step (d); and (f) comparing the binding of the labeled known ligand of said Humanin GPCR to the membrane preparations of step (c) and step (d);

wherein inhibition of binding of the labeled known ligand of said Humanin GPCR in the presence of the compound is indicative of the compound binding to a Humanin GPCR.

In some embodiments, the FPRL2 amino acid sequence is the amino acid sequence of SEQ ID NO:2. In some embodiments, the FPRL2 amino acid sequence is a variant of the amino acid sequence of SEQ ID NO:2. In some embodiments, said variant of the amino acid sequence of SEQ ID NO:2 is an allelic variant or mammalian ortholog of said amino acid sequence. In some embodiments, said variant of the amino acid sequence of SEQ ID NO:2 is a non-endogenous, constitutively activated mutant of said amino acid sequence or of an allelic variant or mammalian ortholog of said amino acid sequence. In certain embodiments, said non-endogenous, constitutively activated mutant is the amino acid sequence of SEQ ID NO:12. In certain embodiments, said variant of the amino acid sequence of SEQ ID NO:2 is a biologically active fragment of said amino acid sequence or of an allelic variant or mammalian ortholog of said amino acid sequence. In certain embodiments, said biologically active fragment is amino acids 2-353 of the amino acid sequence of SEQ HD NO:2 or of an allelic variant or mammalian ortholog of said amino acid sequence. In certain embodiments, said variant of the amino acid sequence of SEQ ID NO:2 is at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% identical to the amino acid sequence of SEQ ID NO:2. In certain embodiments, said variant of the amino acid sequence of SEQ ID NO:2 is at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% identical to the amino acid sequence of SEQ ID NO:2.

In certain embodiments, said membrane preparation is made by homogenization of the cells with a Brinkman Polytron™. In certain embodiments, said membrane preparation is made by homogenization with 3 bursts of 10-20 sec duration each of said polytron.

In certain embodiments, said candidate compound is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof.

In certain embodiments, said candidate compound is not a peptide.

In certain embodiments, said known ligand is Humanin.

In certain embodiments, said known ligand is a peptide analog or derivative of Humanin.

In certain embodiments, said known ligand is [Gly14]-Humanin.

In certain embodiments, said known ligand is Compound 1.

In certain embodiments, said known ligand is a compound according to the second aspect.

In certain embodiments, said known ligand is a compound according to the third aspect.

In certain embodiments, said known ligand is a modulator according to the fourth aspect.

In certain embodiments, said known ligand is an antibody specific for the GPCR, or a derivative of the antibody. In some embodiments, said derivative of the antibody is an antigen-binding fragment of the antibody.

In certain embodiments, said label is selected from the group consisting of:

(a) radioisotope;

(b) enzyme; and

(c) fluorophore.

In certain embodiments, said label is a radioisotope. In certain embodiments, said label is selected from the group consisting of 3 H, 14 C, 35 S, and 125 I.

Radiolabeled Humanin or [Gly14]-Humanin can be chemically synthesized using techniques known in the art [See, e.g., Creighton, 1983 Proteins. New York, N.Y.: W.H. Freeman and Company; and Hunkapiller et al., Nature (1984) 310:105-11]. Radioisotope is introduced through radiolabled amino acid. In certain embodiments, said radiolabled amino acid is 3 H-arginine or 14 C-arginine.

In certain embodiments, said presence of said radiolabeled Humanin or [Gly14]-Humanin is at a concentration of about 0.1 ΣM to about 1.5 μM.

Compound 1 can be radiolabelled using techniques known in the art, infra. In certain embodiments, Compound 1 is radiolabelled with 3 H or 14 C.

In certain embodiments, said presence of said radiolabelled Compound 1 is at a concentration of about 1 μM to about 15 μM.

In other embodiments, said method further comprises the step of comparing the level of inhibition of binding of a labeled first known ligand by the candidate compound to a second level of inhibition of binding of said labeled first known ligand by a second ligand known to bind to the GPCR.

In a seventy-ninth aspect, the invention features a method for detecting ligands that bind to a Humanin GPCR, said receptor comprising an FPRL2 amino acid sequence, comprising the steps of:

contacting a test ligand with a host cell or with membrane of a host cell that expresses said receptor, under conditions which permit interaction between said receptor and said test ligand; and detecting a ligand bound to said receptor.

In some embodiments, the FPRL2 amino acid sequence is the amino acid sequence of SEQ ID NO:2. In some embodiments, the FPRL2 amino acid sequence is a variant of the amino acid sequence of SEQ ID NO:2. In some embodiments, said variant of the amino acid sequence of SEQ ID NO:2 is an allelic variant or mammalian ortholog of said amino acid sequence. In some embodiments, said variant of the amino acid sequence of SEQ ID NO:2 is a non-endogenous, constitutively activated mutant of said amino acid sequence or of an allelic variant or mammalian ortholog of said amino acid sequence. In certain embodiments, said non-endogenous, constitutively activated mutant is the amino acid sequence of SEQ ID NO:12. In certain embodiments, said variant of the amino acid sequence of SEQ ID NO:2 is a biologically active fragment of said amino acid sequence or of an allelic variant or mammalian ortholog of said amino acid sequence. In certain embodiments, said biologically active fragment is amino acids 2-353 of the amino acid sequence of SEQ ID NO:2 or of an allelic variant or mammalian ortholog of said amino acid sequence. In certain embodiments, said variant of the amino acid sequence of SEQ ID NO:2 is at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% identical to the amino acid sequence of SEQ ID NO:2. In certain embodiments, said variant of the amino acid sequence of SEQ ID NO:2 is at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% identical to the amino acid sequence of SEQ ID NO:2.

In certain embodiments, the Humanin GPCR is recombinant. In certain embodiments, said contacting comprises contacting with a host cell or with membrane of a host cell that expresses the GPCR, wherein said host cell comprises an expression vector comprising a polynucleotide encoding the receptor.

In certain embodiments, said candidate compound is not Humanin or an allelic variant, homologue, ortholog, or peptide analog or derivative thereof, or the peptide WKYMVm (SEQ ID NO: 16) or a peptide analog or derivative thereof. In certain embodiments, the modulator is not an antibody or derivative thereof.

In certain embodiments, said test ligand is not a peptide.

In certain embodiments, said membrane preparation is made by homogenization of the cells with a Brinkman Polytron™. In certain embodiments, said membrane preparation is made by homogenization with 3 bursts of 10-20 sec duration each of said polytron.

In certain embodiments, said test ligand is labeled. In certain embodiments, said label is a radioisotope. In certain embodiments, said label is selected from the group consisting of 3 H, 14 C, 35 S and 125 I.

Applicant reserves the right to exclude any one or more candidate compounds from any of the embodiments of the invention. Applicant also reserves the right to exclude any one or more modulators from any of the embodiments of the invention, including but not limited to Humanin or any peptide analog or derivative thereof. Applicant further reserves the right to exclude any polynucleotide or polypeptide from any of the embodiments of the invention. Applicant additionally reserves the right to exclude any cell death-related disorder or any disorder of neuronal cell death or muscle cell death from any of the embodiments of the invention.

Throughout this application, various publications, patents and published patent applications are cited. The disclosures of these publications, patents and published patent applications referenced in this application are hereby incorporated by reference in their entirety into the present disclosure. Citation herein by Applicant of a publication, patent, or published patent application is not an admission by Applicant of said publication, patent, or published patent application as prior art.

Modifications and extension of the disclosed inventions that are within the purview of the skilled artisan are encompassed within the above disclosure and the claims that follow.

FIG. 1 . By way of example and not limitation, FIG. 1 depicts results from a primary screen of candidate compounds against a “target receptor” which is a Gsα Fusion Protein construct of an endogenous, constitutively active Gs-coupled GPCR. Results for “Compound A” are provided in well A2. Results for “Compound “B” are provided in well G9. (See, Example 7.)

FIG. 2 . Microarray analysis was performed on human tissue samples using a custom high-density oligonucleotide microarray, which contains probes that monitor the expression of FPRL2. The histogram plot provides the relative expression levels (Average Difference) and standard errors of duplicate measurements of FPRL2 for each of the tissues profiled. Relative expression level is indicated on the vertical axis. Tissue identity is displayed on the horizontal axis and is (from left to right): Pons, upper; Pituitary gland, female; Sciatic Nerve; Olfactory bulb; Pituitary gland, male; Corpus Callosum, Anterior Hippocampus, Cingulate gyrus, Ventral Tegmental Area (VTA); Neural progenitor; Pons, lower; Spinal cord; Hypothalamus, anterior; Astrocytes, resting; Cerebellum; Frontal Cortex, Superior BM9; Amygdala; Whole brain; Hippocampus; Globus pallidus; Medulla Oblongata; Substantia Nigra; Dorsal Root Ganglion; Thalamus; Astocytes, activated; Fetal Brain; Caudate Putamen; monocytes, adherent; Jurkat; U87; Spleen; Monocytes, CD14+; Thymus; dendritic precursors; Neutrophils; eosinophils; Natural Killer Cells; Bone Marrow; platelets; erythroid progenitors; Lymph Node; T-cells, CD4+ resting; AC133+; T-cells, CD4+ activated; T-cells, CD8+ resting; B-cells, CD19+; T-cells, CD8+ activated; CD34+ progenitor cells; cartilage; Adipocyte, primary; Adipose; Preadipocyte, cultured; Adipocyte, cultured; Pericardium; Aortic Endothelial Cells; Aorta; HUVEC; Aortic Smooth Muscle Cells, proliferative; Aortic smooth muscle cells, contractile; Heart; Rectum; Small Intestine; Colon; Stomach; Liver; Fetal Liver; Lung; Skeletal muscle; Esophagus; Pancreas; Bladder; Thyroid; Trachea; Kidney; Salivary Gland; Skin; Adrenal Gland; pancreatic islets; Mesenchymal stem cell; Bone; Uterus; Prostate; Testis; Ovary; Placenta; Cervix; Breast; Prostate epithelial.

Inspection of the plot indicates that the expression of human FPRL2 encompasses brain, peripheral nerve, and heart. Expression within brain encompasses anterior hippocampus, hippocampus, and substantia nigra. Expression within peripheral nerve encompasses sciatic nerve. Significant FPRL2 expression is also observed, e.g., for spleen, lung, pancreas, bone, and ovary. (See, Example 9.)

FIG. 3 . Analysis of Humanin and [Gly14]-Humanin promoted [ 35 S]GTPγS binding was carried out for human FPRL2 (panel A), FPRL1 (panel B), and FPR (panel C). The EC50 of Humanin at FPRL2 is about 320 nm and that of [Gly14]-Humanin is about 72 nm. These values are more than an order of magnitude lower than the corresponding values at FPRL1, which are about 9.24 μM for Humanin and about 1.5 μM for [Gly14]-Humanin. Neither Humanin nor [Gly14]-Humanin evidence significant activity at FPR up to a dose of 10 μM. Humanin and [Gly14]-Humanin therefore are robust and selective agonists for FPRL2. (See, Example 10.)

FIG. 4 . In melanophores, human FPRL2 couples to Gi in a dose-dependent manner in response to Humanin. The EC50 of Humanin promoted pigment aggregation in melanophore (Gi coupling) is about 239 nM. (See, Example 11.)

FIG. 5 . The activity of Compound 1 (“Cmpd 1” in Table 1) at human FPRL2, FPRL1 and FPR was determined by [ 35 S]GTPγS binding assay and by melanophore assay. Compound 1 was found to be a robust and selective agonist of FPRL2. (See, Example 21.)

FIG. 6 . The activity of Humanin, [Gly14]-Humanin, and Compound 1 at human FPRL2 and FPRL1 was determined by whole cell adenylate cyclase assay. Whereas Human and [Gly14]-Humanin are agonists of both FPRL2 and FPRL1 (panel A), Compound 1 is selectively an agonist of FPRL2 (panel B). (See, Example 21.)

FIG. 7 . The pertussis sensitivity of Humanin signaling through human FPRL2 was determined by adenylate cyclase assay. The pertussis toxin sensitivity of Compound 1 signaling through human FPRL2 was determined similarly. The inhibition of cAMP mediated either by Humanin (panel A) or by Compound 1 (panel B) is pertussis sensitive, indicating that FPRL2 couples to a pertussis-sensitive G protein in response to Humanin or Compound 1. (See, Example 22.)

FIG. 8 . The capacity of FPRL2 to couple to Gal 6 in response to Humanin, [Gly14]-Humanin, or Compound 1 was determined by IP 3 assay. A dose-dependent elevation of the level of intracellular IP 3 by Humanin (panel A), [Gly14]-Humanin (panel B), and Compound 1 (panel C) was observed, indicating that FPRL2 can signal through Gα16 in response to either Humanin or Compound 1. (See, Example 23.)

FIG. 9 . The expression of FPRL2 and FPRL1 by the human neuroblastoma cell line SH-SY5Y, before or after differentiation by brain-derived nerve growth factor (BDNF), was determined. FPRL2 mRNA is detectable both in undifferentiated and differentiated SH-SY5Y cells, whereas FPRL1 mRNA is detectable in neither. (See, Example 24.)

FIG. 10 . The capacity of Humanin or Compound 1 to inhibit the cell death of serum-deprived differentiated SH-SY5Y cells was determined by LDH assay (panel A) or Trypan Blue staining (panel B). Both Humanin and Compound 1 inhibit cell death of serum-deprived SH-SY5Y cells. (See, Example 25.)

FIG. 11 . The capacity of Humanin or Compound 1 to inhibit the cell death of primary mouse cortical neurons induced by anti-APP mAb 22C11 was determined. In panel A is a photograph of a 5 day culture of primary mouse cortical neurons. Humanin and Compound 1 protect primary mouse cortical neurons from cell death induced by anti-APP mAb 22C11 (panel B). (See, Example 26.)

FIG. 12 . The capacity of Humanin or Compound 1 to inhibit the Aβ-induced cell death of differentiated SH-SY5Y cells was determined. Humanin and Compound 1 protect differentiated SH-SY5Y cells from Aβ-induced cell death. (See, Example 27.)

FIG. 13 . An outline of an hypoxia/reoxygenation in vitro model is presented.

FIG. 14 . The expression of FPRL2 in Alzheimer's disease brain relative to that in non-Alzheimer's disease brain was determined by immunohistochemistry. Expression of FPRL2 by the Alzheimer's disease brain sample is up-regulated relative to that in the non-Alzheimer's disease brain sample.

The scientific literature that has evolved around receptors has adopted a number of terms to refer to ligands having various effects on receptors. For clarity and consistency, the following definitions will be used throughout this patent document. To the extent that these definitions conflict with other definitions for these terms, the following definitions shall control:

AGONISTS shall mean materials (e.g., ligands, candidate compounds) that activate an intracelluar response when they bind to the receptor. In some embodiments, AGONISTS are those materials not previously known to activate the intracellular response when they bind to the receptor (e.g. to enhance GTPγS binding to membranes or to lower intracellular cAMP level). In some embodiments, AGONISTS are those materials not previously known to be neuroprotective or myoprotective when they bind to the receptor.

ALLOSTERIC MODULATORS shall mean materials (e.g., ligands, candidate compounds) that affect the functional activity of the receptor but which do not inhibit the endogenous ligand from binding to the receptor. Allosteric modulators include inverse agonists, partial agonists and agonists.

ALZHEIMER'S DISEASE is a progressive neurodegenerative disease characterized by significant loss of function in more than one cognitive domain and often accompanied by changes in behavior or personality. Alzheimer's disease is the most common cause of dementia. The beta-amyloid peptide (Aβ) is believed to play a central role in the pathology of Alzheimer's disease.

AMINO ACID ABBREVIATIONS used herein are set out in Table A:

ANTAGONISTS shall mean materials (e.g., ligands, candidate compounds) that competitively bind to the receptor at the same site as the agonists but which do not activate an intracellular response, and can thereby inhibit the intracellular responses elicited by agonists. ANTAGONISTS do not diminish the baseline intracellular response in the absence of an agonist. In some embodiments, ANTAGONISTS are those materials not previously known to compete with an agonist to inhibit the cellular response when they bind to the receptor, e.g. wherein the cellular response is GTPγS binding to membranes or to the lowering of intracellular cAMP level.

ANTIBODIES are intended herein to encompass monoclonal antibodies and polyclonal antibodies. Antibodies are further intended to encompass IgG, IgA, IgD, IgE, and IgM. Antibodies include whole antibodies, including single-chain whole antibodies, and antigen binding fragments thereof, including Fab, Fab′, F(ab)2 and F(ab′)2. Antibodies may be from any animal origin. Preferably, antibodies are human, murine, rabbit, goat, guinea pig, hamster, camel, donkey, sheep, horse or chicken. Preferably antibodies have binding affinities with a dissociation constant or Kd value less than 5×10 −6 M, 10 −6 M, 5×10 −7 M, 10 −7 M, 5×10 −8 M, 10 −8 M, 5×10 −9 M, 10 −9 M, 5×10 −10 M 10 −10 M, 5×10 −11 M, 10 −11 M, 5×10 −12 M, 10 −12 M, 5×10 −13 M, 10 −13 M, 5×10 −14 M, 10 −14 M, 5×10 −15 M and 10 −15 M. Antibodies of the present invention may be prepared by any suitable method known in the art. Derivatives of antibodies are intended to encompass, but not be limited to, antigen-binding fragments.

BIOLOGICALLY ACTIVE FRAGMENT of a GPCR polypeptide or amino acid sequence shall mean a fragment of the polypeptide or amino acid sequence having structural and biochemical functions of a naturally occurring GPCR. In certain embodiments, the biologically active fragment couples to a G protein. In certain embodiments, the biologically active fragment binds to Humanin.

CANDIDATE COMPOUND shall mean a molecule (for example, and not limitation, a chemical compound) that is amenable to a screening technique.

CEREBRAL AMYLOID BETA-PROTEIN ANGIOPATHY or CEREBRAL AMYLOID ANGIOPATHY (CAA) is a key feature of Alzheimer's disease and related disorders. CAA is a disease of small blood vessels in the brain in which deposits of amyloid protein in the vessel walls may lead to stroke, brain hemorrhage, or dementia. Vascular amyloid deposition leads to degeneration of the vessel wall and aneurysm formation, and may be responsible for 10 to 15% of hemorrhagic strokes in the elderly.

Chemical Group, Moiety or Radical:

The term “C 1-5 acyl” denotes a C 1-4 alkyl radical attached to a carbonyl wherein the definition of alkyl has the same definition as described herein; some examples include but not limited to, acetyl, propionyl, n-butyryl, iso-butyryl, 2-methyl-butyryl, 2,2-dimethyl-propionyl (i.e., pivaloyl), 3-methyl-butyryl and the like.

The term “C 1-5 acyloxy” denotes an C 1-5 acyl radical attached to an oxygen atom wherein C 1-5 acyl has the same definition has described herein; some examples include but not limited to acetyloxy, propionyloxy, n-butyryloxy, iso-butyryloxy, 2-methylbutyryloxy, 2,2-dimethylpropionyloxy (i.e., pivaloyloxy), 3-methylbutyryloxy, and the like.

The term “C 2-6 alkenyl” denotes a radical containing 2 to 6 carbons wherein at least one carbon-carbon double bond is present, some embodiments are 2 to 3 carbons, and some embodiments have 2 carbons. Both E and Z isomers are embraced by the term “alkenyl.” Furthermore, the term “alkenyl” includes di-enes. Accordingly, if more than one double bond is present, then the bonds may be all E or Z or a mixtures of E and Z. Examples of an alkenyl include vinyl, propenyl, allyl, isopropenyl, 2-methyl-propenyl1-methyl-propenyl, but-1-enyl, but-2-enyl, but-3-enyl, buta-1,3-dienyl, and the like.

The term “C 1-4 alkoxy” denotes an alkyl radical, as defined herein, attached directly to an oxygen atom. Examples include methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, t-butoxy, iso-butoxy, sec-butoxy and the like.

The term “C 1-6 alkyl” denotes a straight or branched carbon radical containing the number of carbons as indicated, for examples, in some embodiments, alkyl is a “C 1-4 alkyl” and the group contains 1 to 4 carbons, in still other embodiments, alkyl is a “C 2-6 alkyl” and the group contains 2 to 6 carbons. In some embodiments alkyl contains 1 to 3 carbons, some embodiments contain 1 to 2 carbons, and some embodiments contain 1 carbon. Examples of an alkyl include, but not limited to, methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, t-butyl, sec-butyl, n-pentyl, iso-pentyl, sec-pentyl, neo-pentyl, hexyl, iso-hexyl, sec-hexyl, neo-hexyl, and the like.

The term “C 1-4 alkylcarboxamido” or “C 1-4 alkylcarboxamide” denotes a single C 1-4 alkyl group attached to the nitrogen of an amide group, wherein alkyl has the same definition as found herein. The C 1-5 alkylcarboxamido may be represented by the following:

Examples include, but not limited to, N-methylcarboxamide, N-ethylcarboxamide, N-n-propylcarboxamide, N-iso-propylcarboxamide, N-n-butylcarboxamide, N-sec-butylcarboxamide, N-iso-butylcarboxamide, N-t-butylcarboxamide and the like.

The term “C 2-6 alkynyl” denotes a radical containing 2 to 6 carbons and at least one carbon-carbon triple bond, some embodiments are 2 to 4 carbons, and some embodiments have 2 carbons. Examples of an alkynyl include, but not limited to, ethynyl, prop-1-ynyl, 3-prop-2-ynyl, but-1-ynyl, 1-methyl-prop-2-ynyl, buta-1,3-diynyl, and the like. The term “alkynyl” includes di-ynes.

The term “C 1-4 alkylamino” denotes one alkyl radical attached to an amino radical wherein the alkyl radical has the same meaning as described herein. Some examples include, but not limited to, methylamino, ethylamino, n-propylamino, iso-propylamino, n-butylamino, sec-butylamino, iso-butylamino, t-butylamino, and the like.

The term “C 1-4 alkylsulfonamide” denotes

The term “C 1-4 alkylsulfinyl” denotes a C 1-6 alkyl radical attached to a sulfoxide radical of the formula: —S(O)— wherein the alkyl radical has the same definition as described herein. Examples include, but not limited to, methylsulfinyl, ethylsulfinyl, n-propylsulfinyl, iso-propylsulfinyl, n-butylsulfinyl, sec-butylsulfinyl, iso-butylsulfinyl, t-butyl, and the like.

The term “C 1-4 alkylsulfonyl” denotes a C 1-6 alkyl radical attached to a sulfone radical of the formula: —S(O) 2 — wherein the alkyl radical has the same definition as described herein. Examples include, but not limited to, methylsulfonyl, ethylsulfonyl, n-propylsulfonyl, iso-propylsulfonyl, n-butylsulfonyl, sec-butylsulfonyl, iso-butylsulfonyl, t-butylsulfonyl, and the like.

The term “C 1-4 alkylthio” denotes a C 1-6 alkyl radical attached to a sulfide group of the formula: —S— wherein the alkyl radical has the same definition as described herein. Examples include, but not limited to, methylsulfanyl (i.e., CH 3 S—), ethylsulfanyl, n-propylsulfanyl, iso-propylsulfanyl, n-butylsulfanyl, sec-butylsulfanyl, iso-butylsulfanyl, t-butyl, and the like.

The term “C 1-4 alkylureyl” denotes

The term “amino” denotes the group —NH 2 .

The term “carbo-C 1-6 -alkoxy” denotes a C 1-6 alkyl ester of a carboxylic acid, wherein the alkyl group is as defined herein. Examples include, but not limited to, carbomethoxy, carboethoxy, carbopropoxy, carboisopropoxy, carbobutoxy, carbo-sec-butoxy, carbo-iso-butoxy, carbo-t-butoxy, and the like.

The term “carboxamide” refers to the group —CONH 2 .

The term “carboxy” or “carboxyl” denotes the group —CO 2 H; also referred to as a carboxylic acid group.

The term “cyano” denotes the group —CN.

The term “C 3-6 cycloalkyl” denotes a saturated ring radical containing 3 to 6 carbons; some embodiments, contain 3 to 5 carbons, some embodiments contain 3 to 4 carbons. Examples include, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl.

The term “C 2-6 dialkylamino” denotes an amino substituted with two of the same or different alkyl radicals wherein alkyl radical has the same definition as described herein. A C 2-6 dialkylamino may be represented by the following groups:

Examples of C 2-6 dialkylamino include, but not limited to, dimethylamino, methylethylamino, diethylamino, methylpropylamino, methylisopropylamino, and the like.

The term “C 1-4 dialkylcarboxamido” or “C 1-4 dialkylcarboxamide” denotes two alkyl radicals, that are the same or different, attached to an amide group, wherein alkyl has the same definition as described herein. A C 1-4 dialkylcarboxamido may be represented by the following groups:

wherein C 1-4 has the same definition as described herein. Examples of a dialkylcarboxamide include, but not limited to, N,N-dimethylcarboxamide, N-methyl-N-ethylcarboxamide, N,N-diethylcarboxamide, N-methyl-N-isopropylcarboxamide, and the like.

The term “halogen” or “halo” denotes to a fluoro, chloro, bromo or iodo group.

The term “C 1-4 haloalkoxy” denotes a haloalkyl, as defined herein, which is directly attached to an oxygen atom. Examples include, but not limited to, difluoromethoxy, trifluoromethoxy, 2,2,2-trifluoroethoxy, pentafluoroethoxy and the like.

The term “C 1-4 haloalkyl” denotes an alkyl group wherein the alkyl is substituted with halogen ranging from one to fully substituted, wherein a fully substituted haloalkyl can be represented by the formula C h L 2h+1 wherein L is a halogen and “h” represents the number of carbon atoms; when more than one halogen is present then the halogens may be the same or different and selected from the group consisting of F, Cl, Br and I; it is understood that the terms “alkyl” and “halogen” have the same definition as found herein. In some embodiments, haloalkyl is a “C 1-4 haloalkyl” and the group contains 1 to 4 carbons, some embodiments contain 1 to 3 carbons, some embodiments contain 1 to 2 carbons, some embodiments contain 1 carbon. When the haloalkyl is fully substituted with halogen atoms, this group is referred herein as a perhaloalkyl, one example, is an alkyl fully substituted with fluorine atoms and is referred to herein as a “perfluoroalkyl.” In some embodiments, examples of a haloalkyl include, but not limited to, difluoromethyl, fluoromethyl, 2,2,2-trifluoro-ethyl, 2,2-difluoro-ethyl, 2-fluoro-ethyl, 1,2,2-trifluoro-ethyl, 1,2-difluoro-ethyl, 1,1-difluoro-ethyl, 1,1,2-trifluoro-ethyl, 3,3,3-trifluoro-propyl, 2,2-difluoro-propyl, 3,3-difluoro-propyl, 3-fluoro-propyl, 2,3,3-trifluoro-propyl, 2,3-Difluoro-propyl, 2,2,3,3,3-pentafluoro-propyl, 2,2,3,3-tetrafluoro-propyl, 2,2,3-trifluoro-propyl, 1,2,3,3-tetrafluoro-propyl, 1,2,3-trifluoro-propyl, 3,3-difluoro-propyl, 1,2,2,3-tetrafluoro-propyl, 4,4-difluoro-butyl, 3,3-difluoro-butyl, 4,4,4-trifluoro-butyl, 3,3-difluoro-butyl, and the like. In some embodiments, examples of a perfluoroalkyl include, but not limited to, trifluoromethyl, pentafluoroethyl, heptafluoropropyl, 1,2,2,2-tetrafluoro-1-trifluoromethyl-ethyl, and the like.

The term “C 1-4 haloalkylsulfinyl” denotes a haloalkyl radical attached to a sulfoxide group of the formula: —S(O)— wherein the haloalkyl radical has the same definition as described herein.

The term “C 1-4 haloalkylsulfonyl” denotes a haloalkyl radical attached to a sulfone group of the formula: —S(O) 2 — wherein haloalkyl has the same definition as described herein.

The term “C 1-4 haloalkylthio” denotes a haloalkyl radical directly attached to a sulfur atom wherein the haloalkyl has the same meaning as described herein.

The term “hydroxyl” denotes the group —OH.

The term “nitro” denotes the group —NO 2 .

CODON shall mean a grouping of three nucleotides (or equivalents to nucleotides) which generally comprise a nucleoside [adenosine (A), guanosine (G), cytidine (C), uridine (U) and thymidine (T)] coupled to a phosphate group and which, when translated, encodes an amino acid.

COMPOSITION means a material comprising at least one component. A “pharmaceutical composition” is an example of a composition.

COMPOUND EFFICACY shall mean a measurement of the ability of a compound to inhibit or stimulate receptor functionality; i.e. the ability to activate/inhibit a signal transduction pathway, in contrast to receptor binding affinity. Exemplary means of detecting compound efficacy are disclosed in the Example section of this patent document.

COMPRISING, CONSISTING ESSENTIALLY OF, and CONSISTING OF are defined herein according to their standard meaning. A defined meaning set forth in the M.P.E.P. controls over a defined meaning in the art and a defined meaning set forth in controlling Federal Circuit case law controls over a meaning set forth in the M.P.E.P.

CONGESTIVE HEART FAILURE shall refer to a disorder in which the heart loses its ability to pump blood efficiently. Congestive heart failure becomes more prevalent with advancing age. Ischemic heart disease is the most common cause of congestive heart failure, accounting for 60-70% of all cases. An increased venous pressure greater than 12 mmHg is one of the major Framingham criteria for congestive heart failure, as is a reduction in cardiac output equivalent to a circulation time greater than 25 seconds.

CONSTITUTIVELY ACTIVE RECEPTOR shall mean a receptor stabilized in an active state by means other than through binding of the receptor to its ligand or a chemical equivalent thereof A CONSTITUTIVELY ACTIVE RECEPTOR may be endogenous or non-endogenous.

CONSTITUTIVELY ACTIVATED RECEPTOR shall mean an endogenous receptor that has been modified so as to be constitutively active.

CONSTITUTIVE RECEPTOR ACTIVATION shall mean activation of a receptor in the absence of binding to its ligand or a chemical equivalent thereof.

CONTACT or CONTACTING shall mean bringing at least two moieties together, whether in an in vitro system or an in vivo system.

DECREASE is used to refer to a reduction in a measurable quantity and is used synonymously with the terms “reduce”, “diminish”, “lower”, and “lessen”.

ENDOGENOUS shall mean a material that a mammal naturally produces. ENDOGENOUS in reference to, for example and not limitation, the term “receptor,” shall mean that which is naturally produced by a mammal (for example, and not limitation, a human). ENDOGENOUS shall be understood to encompass allelic variants of a gene as well as the allelic polypeptide variants so encoded. As used herein, “endogenous GPCR” and “native GPCR” are used interchangeably. By contrast, the term NON-ENDOGENOUS in this context shall mean that which is not naturally produced by a mammal (for example, and not limitation, a human). For example, and not limitation, a receptor which is not constitutively active in its endogenous form, but when manipulated becomes constitutively active, is most preferably referred to herein as a “non-endogenous, constitutively activated receptor.”

EXPRESSION VECTOR is defined herein as a DNA sequence that is required for the transcription of cloned DNA and the translation of the transcribed mRNAs in an appropriate host cell recombinant for said EXPRESSION VECTOR. An appropriately constructed EXPRESSION VECTOR should contain an origin of replication for autonomous replication in host cells, selectable markers, a limited number of useful restriction enzyme sites, a potential for high copy number, and active promoters. Said cloned DNA to be transcribed is operably linked to a constitutively or conditionally active promoter within said expression vector. By way of illustration and not limitation, pCMV is an expression vector.

G PROTEIN COUPLED RECEPTOR FUSION PROTEIN and GPCR FUSION PROTEIN, in the context of the invention disclosed herein, each mean a non-endogenous protein comprising an endogenous, constitutively active GPCR or a non-endogenous, constitutively activated GPCR fused to at least one G protein, most preferably the alpha (α) subunit of such G protein (this being the subunit that binds GTP), with the G protein preferably being of the same type as the G protein that naturally couples with endogenous GPCR. For example, and not limitation, in an endogenous state, if the G protein “G s α” is the predominate G protein that couples with the GPCR, a GPCR Fusion Protein based upon the specific GPCR would be a non-endogenous protein comprising the GPCR fused to G s α; in some circumstances, as will be set forth below, a non-predominant G protein can be fused to the GPCR. The G protein can be fused directly to the C-terminus of the constitutively active GPCR or there may be spacers between the two.

HOST CELL shall mean a cell capable of having a vector incorporated therein. In certain embodiments, the vector is an expression vector. Exemplary host cells include but are not limited to 293, 293T, CHO, MCB3901, and COS-7 cells, as well as melanophore cells.

HUMANIN ACTIVITY as relates to a compound shall mean said compound binds to and is a modulator of a Humanin receptor, wherein said receptor preferably is endogenous and more preferably FPRL2, and wherein said modulator preferably is agonist.

IN NEED OF PREVENTION OR TREATMENT as used herein refers to a judgement made by a caregiver (e.g. physician, nurse, nurse practitioner, etc. in the case of humans; veterinarian in the case of animals, including non-human mammals) that an individual or animal requires or will benefit from treatment. This judgement is made based on a variety of factors that are in the realm of a caregiver's expertise, but that include the knowledge that the individual or animal is ill, or will be ill, as the result of a condition that is treatable by the compounds of the invention.

INDIVIDUAL as used herein refers to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, swine, cattle, sheep, horses, or primates, and most preferably humans.

INHIBIT or INHIBITING, in relationship to the term “response” shall mean that a response is decreased or prevented in the presence of a compound as opposed to in the absence of the compound.

INVERSE AGONISTS shall mean materials (e.g., ligand, candidate compound) that bind either to the endogenous form or to the constitutively activated form of the receptor so as to reduce the baseline intracellular response of the receptor observed in the absence of agonists.

ISCHEMIC HEART DISEASE shall refer to a disorder caused by lack of oxygen to the tissues of the heart, in which muscles of the heart are affected and the heart cannot pump properly. Ischemic heart disease is the most common cardiomyopathy in the United States.

ISOLATED shall mean that the material is removed from its original environment (e.g., the natural environment if it is naturally occurring). For example, a naturally occurring polynucleotide or polypeptide present in a living animal is not isolated, but the same polynucleotide or DNA or polypeptide, separated from some or all of the coexisting materials in the natural system, is isolated. Such a polynucleotide could be part of a vector and/or such a polynucleotide or polypeptide could be part of a composition, and still be isolated in that the vector or composition is not part of its natural environment.

LIGAND shall mean a molecule that specifically binds to a GPCR. A ligand may be, for example, a polypeptide, a lipid, a small molecule, an antibody. An endogenous ligand is a ligand that is an endogenous, natural ligand for a native GPCR. A ligand may be a GPCR “antagonist”, “agonist”, “partial agonist”, or “inverse agonist”, or the like.

As used herein, the terms MODULATE or MODIFY are meant to refer to an increase or decrease in the amount, quality, or effect of a particular activity, function or molecule. By way of illustration and not limitation, agonists, partial agonists, inverse agonists, and antagonists of a G protein-coupled receptor are modulators of the receptor.

MOTOR NEURON DISEASE shall refer to a disease characterized by a selective degeneration of the motor neurons of the spinal cord, brainstem, or motor cortex. Clinical subtypes are distinguished by the major site of degeneration.

MYOCARDIAL INFARCTION shall refer to the damage or death of an area of heart muscle because of an inadequate supply of oxygen to that area. Myocardial infarctions are often caused by a clot that blocks one of the coronary arteries (the blood vessels that bring blood and oxygen to heart muscle). The clot prevents blood and oxygen from reaching that area of the heart, leading to the death of heart cells in that area.

MYOPROTECTIVE shall refer to the property of protecting against muscle cell death in one or more contexts. It follows that a modulator of a myoprotective GPCR is a modulator of myoprotection.

NEUROPROTECTIVE shall refer to the property of protecting against neuronal cell death in one or more contexts. It follows that a modulator of a neuroprotective GPCR is a modulator of neuroprotection.

PARKINSON'S DISEASE is a chronic, progressive neurodegenerative disorder characterized by motor symptoms such as tremor, bradykinesia, muscle rigidity, gait dysfunction, and postural instability. Although researchers have identified degeneration of dopaminergic neurons in the substantia nigra as the primary pathophysiological mechanism, they believe that other neurotransmitter systems—such as the serontonergic and glutamatergic systems—are also intricately involved in the disease process.

PARTIAL AGONISTS shall mean materials (e.g. ligands, candidate compounds) that activate the intracellular response when they bind to the receptor to a lesser degree/extent than do full agonists.

PERIPHERAL NEUROPATHY is a broad term used to describe a variety of disorders which manifest as painful or uncomfortable sensations usually in the extremities and are of neuropathic origin. This condition is found commonly amongst end stage renal disease and diabetic patients. Distal symmetrical polyneuropathy (DSP)—a neuropathy affecting both diodes of the body and attacking the distal sensory nerves—is the most common clinical manifestation of diabetic neuropathy and accounts for approximately 80-85% of all neuropathy observed in diabetic patients.

PHARMACEUTICAL COMPOSITION shall mean a composition comprising at least one active ingredient, whereby the composition is amenable to investigation for a specified, efficacious outcome in a mammal (for example, and not limitation, a human). Those of ordinary skill in the art will understand and appreciate the techniques appropriate for determining whether an active ingredient has a desired efficacious outcome based upon the needs of the artisan.

POLYNUCLEOTIDES shall mean RNA, DNA, or RNA/DNA hybrid sequences of more than one nucleotide in either single chain or duplex form. The polynucleotides of the invention may be prepared by any known method, including synthetic, recombinant, ex vivo generation, or a combination thereof, as well as utilizing any purification methods known in the art.

POLYPEPTIDE shall refer to a polymer of amino acids without regard to the length of the polymer. Thus, PEPTIDES, oligopeptides, and proteins are included within the definition of polypeptide. This term also does not specify or exclude post-expression modifications of polypeptides. For example, polypeptides that include the covalent attachment of glycosyl groups, acetyl groups, phosphate groups, lipid groups and the like are expressly encompassed by the term POLYPEPTIDE.

PRIMER is used herein to denote a specific oligonucleotide sequence which is complementary to a target nucleotide sequence and used to hybridize to the target nucleotide sequence. A primer serves as an initiation point for nucleotide polymerization catalyzed by DNA polymerase, RNA polymerase, or reverse transcriptase.

PRION-ASSOCIATED DISEASE is intended herein to encompass, but not be limited to, bovine spongiform encephalopathy, Creutzfeldt-Jakob disease, kuru, Gerstmann-Straussler-Scheinker syndrome, and fatal familial insomnia.

PURIFIED is used herein to describe a polynucleotide or polynucleotide vector of the invention that has been separated from other compounds including, but not limited to, other nucleic acids, carbohydrates, lipids and proteins (such as the enzymes used in the synthesis of the polynucleotide). In certain embodiments, a polynucleotide is substantially pure when at least about 50%, at least about 60%, at least about 75%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% of a sample contains a single polynucleotide sequence. In some embodiments, a substantially pure polynucleotide typically comprises about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99% or about 99.5% weight/weight of a polynucleotide sample.

Similarly, the term PURIFIED is used herein to describe a polypeptide of the invention that has been separated from other compounds including, but not limited to, nucleic acids, lipids, carbohydrates and other proteins. In certain embodiments, a polypeptide is substantially pure when at least about 50%, at least about 60%, at least about 75%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% of the polypeptide molecules of a sample have a single amino acid sequence. In some embodiments, a substantially pure polypeptide typically comprises about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99% or about 99.5% weight/weight of a protein sample.

Similarly, the term PURIFIED is used herein to describe a modulator of the invention. In certain embodiments, a substantially pure modulator typically comprises at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or at least about 99.5% weight/weight of a preparation of said modulator. In certain embodiments, the modulator has an “at least” purity ranging from any number, to the thousandth position, between 90% and 100% (e.g., at least 99.995% pure).

Further, as used herein, the term PURIFIED does not require absolute purity; rather, it is intended as a relative definition.

RECEPTOR FUNCTIONALITY shall refer to the normal operation of a receptor to receive a stimulus and moderate an effect in the cell, including, but not limited to regulating gene transcription, regulating the influx or efflux of ions, effecting a catalytic reaction, and/or modulating activity through G-proteins.

SECOND MESSENGER shall mean an intracellular response produced as a result of receptor activation. A second messenger can include, for example, inositol triphosphate (IP 3 ), diacylglycerol (DAG), cyclic AMP (cAMP), cyclic GMP (cGMP), and Ca 2+ . Second messenger response can be measured for a determination of receptor activation. In addition, second messenger response can be measured for the identification of candidate compounds as, for example, inverse agonists, partial agonists, agonists, and antagonists.

SIGNAL TO NOISE RATIO shall mean the signal generated in response to activation, amplification, or stimulation wherein the signal is above the background noise or the basal level in response to non-activation, non-amplification, or non-stimulation.

SPACER shall mean a translated number of amino acids that are located after the last codon or last amino acid of a gene, for example a GPCR of interest, but before the start codon or beginning regions of the G protein of interest, wherein the translated number amino acids are placed in-frame with the beginnings regions of the G protein of interest. The number of translated amino acids can be one, two, three, four, etc., and up to twelve.

STIMULATE or STIMULATING, in relationship to the term “response” shall mean that a response is increased in the presence of a compound as opposed to in the absence of the compound.

STROKE is a cardiovascular disease that affects the blood vessels supplying blood to the brain and is intended herein to include cerebral thrombosis, the most common type of STROKE. Cerebral thrombosis occurs when a blood clot (thrombus) forms and blocks blood flow in an artery bringing blood to part of the brain. Blood clots usually form in arteries damaged by atherosclerosis.

SUBJECT shall mean primates, including but not limited to humans and baboons, as well as pet animals such as dogs and cats, laboratory animals such as rats and mice, and farm animals such as horses, sheep, and cows.

THERAPEUTICALLY EFFECTIVE AMOUNT as used herein refers to the amount of active compound or pharmaceutical agent that elicits the biological or medicinal response in a tissue, system, animal, individual or human that is being sought by a researcher, veterinarian, medical doctor or other clinician, which includes one or more of the following:

(1) Preventing the disease; for example, preventing a disease, condition or disorder in an individual that may be predisposed to the disease, condition or disorder but does not yet experience or display the pathology or symptomatology of the disease, (2) Inhibiting the disease; for example, inhibiting a disease, condition or disorder in an individual that is experiencing or displaying the pathology or symptomatology of the disease, condition or disorder (i.e., arresting further development of the pathology and/or symptomatology), and (3) Ameliorating the disease; for example, ameliorating a disease, condition or disorder in an individual that is experiencing or displaying the pathology or symptomatology of the disease, condition or disorder (i.e., reversing the pathology and/or symptomatology).

VARIANT as the term is used herein, is a polynucleotide or polypeptide that differs from a reference polynucleotide or polypeptide respectively, but retains essential properties. A typical variant of a polynucleotide differs in nucleotide sequence from another, reference polynucleotide. Changes in the nucleotide sequence of the variant may or may not alter the amino acid sequence of a polypeptide encoded by the reference polynucleotide. A typical variant of a polypeptide differs in amino acid sequence from another, reference polypeptide. A variant and reference polypeptide may differ in amino acid sequence by one or more substitutions, additions, deletions in any combination. A variant of a polynucleotide or polypeptide may be a naturally occurring one such as an ALLELIC VARIANT, or it may be a variant that is not known to occur naturally. Non-naturally occurring variants of polynucleotides and polypeptides may be made by mutagenesis techniques or by direct synthesis.

A. Introduction

The order of the following sections is set forth for presentational efficiency and is not intended, nor should be construed, as a limitation on the disclosure or the claims to follow.

B. Receptor Expression

1. GPCR Polypeptides of Interest

As used herein, “an FPRL2 amino acid sequence” is intended to encompass the endogenous human FPRL2 amino acid sequence of SEQ ID NO:2 as well as a variant amino acid sequence at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% identical to the amino acid sequence of SEQ ID NO:2. In other words, a GPCR comprising a variant of the amino acid sequence of SEQ ID NO:2 also may be used in the subject methods. In certain embodiments, a GPCR that may be used in the subject methods may comprise an allelic variant of the amino acid sequence of SEQ ID NO:2. By way of illustration and not limitation, an allelic variant of the amino acid sequence of SEQ ID NO:2 may comprise a substitution of alanine for glycine at amino acid position 94 of SEQ ID NO:2, a substitution of threonine for serine at amino acid position 211 of SEQ ID NO:2, a substitution of histidine for aspartic acid at amino acid position 338 of SEQ ID NO:2, or may comprise any combination of said amino acid substitutions (GenBank® Accession No. AAA58482). In certain embodiments, an allelic variant of the amino acid sequence of SEQ ID NO:2 is encoded by an endogenous FPRL2 nucleotide sequence obtainable by performing polymerase chain reaction (PCR) on a human DNA sample using specific primers SEQ ID NO:3 and SEQ ID NO:4. In some embodiments, an allelic variant of the amino acid sequence of SEQ ID NO:2 is encoded by an endogenous FPRL2 nucleotide sequence obtainable by performing polymerase chain reaction (PCR) on a human DNA sample using a specific primer comprising SEQ ID NO:3 and a specific primer comprising SEQ ID NO:4. In certain embodiments, a variant amino acid sequence that may be used in the subject methods is a mammalian ortholog of the amino acid sequence of SEQ ID NO:2. By way of illustration and not limitation, the FPRL2 amino acid sequences of Pan troglodytes (GenBank® Accession No. P79243, e.g.), Gorilla gorilla gorilla (GenBank® Accession No. P79178, e.g.), Pongo pygmaeus (GenBank® Accession No. P79237, e.g.), and Macaca mulatto (GenBank® Accession No. P79191) are envisioned to be within the scope of “an FPRL2 amino acid sequence”. In certain embodiments, a GPCR that may be used in the subject methods may comprise a non-endogenous, constitutively activated mutant of the amino acid sequence of SEQ ID NO:2, an allele of SEQ ID NO:2, or a mammalian ortholog of SEQ ID NO:2. As is known in the art, a constitutively activated GPCR may be made using a variety of methods (see, e.g., PCT Application Number PCT/US98/07496 published as WO 98/46995 on 22 Oct. 1998; and U.S. Pat. No. 6,555,339; the disclosure of each of which is hereby incorporated by reference in its entirety.) A biologically active fragment of the amino acid sequence of SEQ ID NO:2, of an allele of SEQ ID NO:2, of a mammalian ortholog of SEQ ID NO:2, of a non-endogenous, constitutively activated mutant of endogenous FPRL2, or of an amino acid sequence at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% identical to the amino acid sequence of SEQ ID NO:2 may be used in the subject invention. By way of illustration and not limitation, deletion of an N-terminal methionine or an N-terminal signal peptide is envisioned to provide a biologically active fragment that may be used in the subject methods.

In certain embodiments, a GPCR that may be used in the subject methods may comprise an amino acid sequence at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% identical to the amino acid sequence of SEQ ID NO:2. In certain embodiments, a GPCR that may be used in the subject methods may comprise an amino acid sequence at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99% identical to the amino acid sequence of SEQ ID NO:2. In certain embodiments, a GPCR that may be used in the subject methods may comprise an amino acid sequence at least about 75% identical to the amino acid sequence of SEQ ID NO:2. In certain embodiments, a GPCR that may be used in the subject methods may comprise an amino acid sequence at least about 80% identical to the amino acid sequence of SEQ ID NO:2. In certain embodiments, a GPCR that may be used in the subject methods may comprise an amino acid sequence at least about 85% identical to the amino acid sequence of SEQ ID NO:2. In certain embodiments, a GPCR that may be used in the subject methods may comprise an amino acid sequence at least about 90% identical to the amino acid sequence of SEQ ID NO:2. In certain embodiments, a GPCR that may be used in the subject methods may comprise an amino acid sequence at least about 91% identical to the amino acid sequence of SEQ ID NO:2. In certain embodiments, a GPCR that may be used in the subject methods may comprise an amino acid sequence at least about 92% identical to the amino acid sequence of SEQ ID NO:2. In certain embodiments, a GPCR that may be used in the subject methods may comprise an amino acid sequence at least about 93% identical to the amino acid sequence of SEQ ID NO:2. In certain embodiments, a GPCR that may be used in the subject methods may comprise an amino acid sequence at least about 94% identical to the amino acid sequence of SEQ ID NO:2. In certain embodiments, a GPCR that may be used in the subject methods may comprise an amino acid sequence at least about 95% identical to the amino acid sequence of SEQ ID NO:2. In certain embodiments, a GPCR that may be used in the subject methods may comprise an amino acid sequence at least about 96% identical to the amino acid sequence of SEQ ID NO:2. In certain embodiments, a GPCR that may be used in the subject methods may comprise an amino acid sequence at least about 97% identical to the amino acid sequence of SEQ ID NO:2. In certain embodiments, a GPCR that may be used in the subject methods may comprise an amino acid sequence at least about 98% identical to the amino acid sequence of SEQ ID NO:2. In certain embodiments, a GPCR that may be used in the subject methods may comprise an amino acid sequence at least about 99% identical to the amino acid sequence of SEQ ID NO:2. By an amino acid sequence having at least, for example, 95% “identity” to the amino acid sequence of SEQ ID NO:2 is meant that the amino acid sequence is identical to the amino acid sequence of SEQ ID NO:2 except that it may include up to five amino acid alterations per each 100 amino acids of the amino acid sequence of SEQ ID NO:2. Thus, to obtain an amino acid sequence having at least 95% identity to that of SEQ ID NO:2, up to 5% (5 of 100) of the amino acid residues in the sequence may be inserted, deleted, or substituted with another amino acid compared with the amino acid sequence of SEQ ID NO:2. These alternations may occur at the amino or carboxy termini or anywhere between those terminal positions, interspersed either individually among residues in the sequence or in one or more contiguous groups within the sequence.

In some embodiments, an FPRL2 amino acid sequence that may be used in the subject methods is an amino acid sequence of a G protein-coupled receptor encoded by a complementary sequence to the sequence of a polynucleotide that hybridizes under stringent conditions to filter-bound DNA having the sequence set forth in SEQ ID NO:1. Hybridization techniques are well known to the skilled artisan. Preferred stringent hybridization conditions include overnight incubation at 42° C. in a solution comprising: 50% formamide, 5×SSC (150 mM NaCl, 15 mM trisodium citrate), 5 mM sodium phosphate (pH 7.6), 5×Denhardt's solution, 10% dextran sulfate, and 20 g/ml denatured, sheared salmon sperm DNA; followed by washing the filter in 0.1×SSC at about 65° C.

a. Sequence Identity

A preferred method for determining the best overall match between a query sequence (e.g., the amino acid sequence of SEQ ID NO:2) and a sequence to be interrogated, also referred to as a global sequence alignment, can be determined using the FASTDB computer program based on the algorithm of Brutlag et al. [Comp App Biosci (1990) 6:237-245; the disclosure of which is hereby incorporated by reference in its entirety]. In a sequence alignment the query and interrogated sequences are both amino acid sequences. The results of said global sequence alignment is in percent identity. Preferred parameters used in a FASTDB amino acid alignment are: Matrix=PAM 0, k-tuple=2, Mismatch Penalty=1, Joining Penalty=20, Randomization Group=25, Length=0, Cutoff Score=1, Window Size=sequence length, Gap Penalty=5, Gap Size Penalty=0.05, Window Size=247 or the length of the interrogated amino acid sequence, whichever is shorter.

If the interrogated sequence is shorter than the query sequence due to N- or C-terminal deletions, not because of internal deletions, the results, in percent identity, must be manually corrected because the FASTDB program does not account for N- and C-terminal truncations of the interrogated sequence when calculating global percent identity. For interrogated sequences truncated at the N- and C-termini, relative to the query sequence, the percent identity is corrected by calculating the number of residues of the query sequence that are N- and C-terminal of the interrogated sequence, that are not matched/aligned with a corresponding interrogated sequence residue, as a percent of the total bases of the query sequence. Whether a residue is matched/aligned is determined by results of the FASTDB sequence alignment. This percentage is then subtracted from the percent identity, calculated by the above FASTDB program using the specified parameters, to arrive at a final percent identity score. This final percent identity score is what is used for the purposes of the present invention. Only residues to the N- and C-termini of the interrogated sequence, which are not matched/aligned with the query sequence, are considered for the purposes of manually adjusting the percent identity score. That is, only query amino acid residues outside the farthest N- and C-terminal residues of the interrogated sequence.

For example, a 90 amino acid residue interrogated sequence is aligned with a 100-residue query sequence to determine percent identity. The deletion occurs at the N-terminus of the interrogated sequence and therefore, the FASTDB alignment does not match/align with the first residues at the N-terminus. The 10 unpaired residues represent 10% of the sequence (number of residues at the N- and C-termini not matched/total number of residues in the query sequence) so 10% is subtracted from the percent identity score calculated by the FASTDB program. If the remaining 90 residues were perfectly matched, the final percent identity would be 90%.

In another example, a 90-residue interrogated sequence is compared with a 100-residue query sequence. This time the deletions are internal so there are no residues at the N- or C-termini of the interrogated sequence, which are not matched/aligned with the query. In this case, the percent identity calculated by FASTDB is not manually corrected. Once again, only residue positions outside the N- and C-terminal ends of the subject sequence, as displayed in the FASTDB alignment, which are not matched/aligned with the query sequence are manually corrected. No other corrections are made for the purposes of the present invention.

b. Fusion Proteins

In certain embodiments, a polypeptide of interest is a fusion protein, and may contain, for example, an affinity tag domain or a reporter domain. Suitable affinity tags include any amino acid sequence that may be specifically bound to another moiety, usually another polypeptide, most usually an antibody. Suitable affinity tags include epitope tags, for example, the V5 tag, the FLAG tag, the HA tag (from hemagglutinin influenza virus), the myc tag, and the like, as is known in the art. Suitable affinity tags also include domains for which, binding substrates are known, e.g., HIS, GST and MBP tags, as is known in the art, and domains from other proteins for which specific binding partners, e.g., antibodies, particularly monoclonal antibodies, are available. Suitable affinity tags also include any protein-protein interaction domain, such as a IgG Fc region, which may be specifically bound and detected using a suitable binding partner, e.g. the IgG Fc receptor. It is expressly contemplated that such a fusion protein may contain a heterologous N-terminal domain (e.g., an epitope tag) fused in-frame with a GPCR that has had its N-terminal methionine residue either deleted or substituted with an alternative amino acid.

Suitable reporter domains include any domain that can report the presence of a polypeptide. While it is recognized that an affinity tag may be used to report the presence of a polypeptide using, e.g., a labeled antibody that specifically binds to the tag, light emitting reporter domains are more usually used. Suitable light emitting reporter domains include luciferase (from, e.g., firefly, Vargula, Renilla reniformis or Renilla muelleri ), or light emitting variants thereof. Other suitable reporter domains include fluorescent proteins, (from e.g., jellyfish, corals and other coelenterates as such those from Aequoria, Renilla, Ptilosarcus, Stylatula species), or light emitting variants thereof. Light emitting variants of these reporter proteins are very well known in the art and may be brighter, dimmer, or have different excitation and/or emission spectra, as compared to a native reporter protein. For example, some variants are altered such that they no longer appear green, and may appear blue, cyan, yellow, enhanced yellow red (termed BFP, CFP, YFP eYFP and RFP, respectively) or have other emission spectra, as is known in the art. Other suitable reporter domains include domains that can report the presence of a polypeptide through a biochemical or color change, such as β-galactosidase, β-glucuronidase, chloramphenicol acetyl transferase, and secreted embryonic alkaline phosphatase.

Also as is known in the art, an affinity tags or a reporter domain may be present at any position in a polypeptide of interest. However, in most embodiments, they are present at the C- or N-terminal end of a polypeptide of interest.

2. Nucleic Acids Encoding GPCR Polypeptides of Interest

Since the genetic code and recombinant techniques for manipulating nucleic acid are known, and the amino acid sequences of GPCR polypeptides of interest described as above, the design and production of nucleic acids encoding a GPCR polypeptide of interest is well within the skill of an artisan. In certain embodiments, standard recombinant DNA technology (Ausubel, et al, Short Protocols in Molecular Biology, 3rd ed., Wiley & Sons, 1995; Sambrook, et al., Molecular Cloning: A Laboratory Manual , Second Edition, (1989) Cold Spring Harbor, N.Y.) methods are used. For example, GPCR coding sequences may be isolated from a library of GPCR coding sequence using any one or a combination of a variety of recombinant methods that do not need to be described herein. Subsequent substitution, deletion, and/or addition of nucleotides in the nucleic acid sequence encoding a protein may also be done using standard recombinant DNA techniques.

For example, site directed mutagenesis and subcloning may be used to introduce/delete/substitute nucleic acid residues in a polynucleotide encoding a polypeptide of interest. In other embodiments, PCR may be used. Nucleic acids encoding a polypeptide of interest may also be made by chemical synthesis entirely from oligonucleotides (e.g., Cello et al., Science (2002) 297:1016-8).

In some embodiments, the codons of the nucleic acids encoding polypeptides of interest are optimized for expression in cells of a particular species, particularly a mammalian, e.g., mouse, rat, hamster, non-human primate, or human, species. In some embodiments, the codons of the nucleic acids encoding polypeptides of interest are optimized for expression in cells of a particular species, particularly an amphibian species.

a. Vectors

The invention further provides vectors (also referred to as “constructs”) comprising a subject nucleic acid. In many embodiments of the invention, the subject nucleic acid sequences will be expressed in a host after the sequences have been operably linked to an expression control sequence, including, e.g. a promoter. The subject nucleic acids are also typically placed in an expression vector that can replicate in a host cell either as an episome or as an integral part of the host chromosomal DNA. Commonly, expression vectors will contain selection markers, e.g., tetracycline or neomycin, to permit detection of those cells transformed with the desired DNA sequences (see, e.g., U.S. Pat. No. 4,704,362, which is incorporated herein by reference). Vectors, including single and dual expression cassette vectors are well known in the art (Ausubel, et al, Short Protocols in Molecular Biology, 3rd ed., Wiley & Sons, 1995; Sambrook, et al., Molecular Cloning: A Laboratory Manual , Second Edition, (1989) Cold Spring Harbor, N.Y.). Suitable vectors include viral vectors, plasmids, cosmids, artificial chromosomes (human artificial chromosomes, bacterial artificial chromosomes, yeast artificial chromosomes, etc.), mini-chromosomes, and the like. Retroviral, adenoviral and adeno-associated viral vectors may be used.

A variety of expression vectors are available to those in the art for purposes of producing a polypeptide of interest in a cell. One suitable vector is pCMV, which is used in certain embodiments. This vector was deposited with the American Type Culture Collection (ATCC) on Oct. 13, 1998 (10801 University Blvd., Manassas, Va. 20110-2209 USA) under the provisions of the Budapest Treaty for the International Recognition of the Deposit of Microorganisms for the Purpose of Patent Procedure. The DNA was tested by the ATCC and determined to be viable. The ATCC has assigned the following deposit number to pCMV: ATCC #203351.

The subject nucleic acids usually comprise an single open reading frame encoding a subject polypeptide of interest, however, in certain embodiments, since the host cell for expression of the polypeptide of interest may be a eukaryotic cell, e.g., a mammalian cell, such as a human cell, the open reading frame may be interrupted by introns. Subject nucleic acid are typically part of a transcriptional unit which may contain, in addition to the subject nucleic acid 3′ and 5′ untranslated regions (UTRs) which may direct RNA stability, translational efficiency, etc. The subject nucleic acid may also be part of an expression cassette which contains, in addition to the subject nucleic acid a promoter, which directs the transcription and expression of a polypeptide of interest, and a transcriptional terminator.

Eukaryotic promoters can be any promoter that is functional in a eukaryotic host cell, including viral promoters and promoters derived from eukaryotic genes. Exemplary eukaryotic promoters include, but are not limited to, the following: the promoter of the mouse metallothionein I gene sequence (Hamer et al., J. Mol. Appl. Gen. 1:273-288, 1982); the TK promoter of Herpes virus (McKnight, Cell 31:355-365, 1982); the SV40 early promoter (Benoist et al., Nature (London) 290:304-310, 1981); the yeast gall gene sequence promoter (Johnston et al., Proc. Natl. Acad. Sci. (USA) 79:6971-6975, 1982); Silver et al., Proc. Natl. Acad. Sci. (USA) 81:5951-59SS, 1984), the CMV promoter, the EF-1 promoter, Ecdysone-responsive promoter(s), tetracycline-responsive promoter, and the like. Viral promoters may be of particular interest as they are generally particularly strong promoters. In certain embodiments, a promoter is used that is a promoter of the target pathogen. Promoters for use in the present invention are selected such that they are functional in the cell type (and/or animal) into which they are being introduced. In certain embodiments, the promoter is a CMV promoter.

In certain embodiments, a subject vector may also provide for expression of a selectable marker. Suitable vectors and selectable markers are well known in the art and discussed in Ausubel, et al, (Short Protocols in Molecular Biology, 3rd ed., Wiley & Sons, 1995) and Sambrook, et al, (Molecular Cloning: A Laboratory Manual, Third Edition, (2001) Cold Spring Harbor, N.Y.). A variety of different genes have been employed as selectable markers, and the particular gene employed in the subject vectors as a selectable marker is chosen primarily as a matter of convenience. Known selectable marker genes include: the thymidine kinase gene, the dihydrofolate reductase gene, the xanthine-guanine phosphoribosyl transferase gene, CAD, the adenosine deaminase gene, the asparagine synthetase gene, the antibiotic resistance genes, e.g. tetr, ampr, Cmr or cat, kanr or neor (aminoglycoside phosphotransferase genes), the hygromycin B phosphotransferase gene, and the like.

As mentioned above, polypeptides of interest may be fusion proteins that contain an affinity domain and/or a reporter domain. Methods for making fusions between a reporter or tag and a GPCR, for example, at the C- or N-terminus of the GPCR, are well within the skill of one of skill in the art (e.g. McLean et al, Mol. Pharma. Mol Pharmacol. 1999 56:1182-91; Ramsay et al., Br. J. Pharmacology, 2001, 315-323) and will not be described any further. It is expressly contemplated that such a fusion protein may contain a heterologous N-terminal domain (e.g., an epitope tag) fused in-frame with a GPCR that has had its N-terminal methionine residue either deleted or substituted with an alternative amino acid. It is appreciated that a polypeptide of interest may first be made from a native polypeptide and then operably linked to a suitable reporter/tag as described above.

The subject nucleic acids may also contain restriction sites, multiple cloning sites, primer binding sites, ligatable ends, recombination sites etc., usually in order to facilitate the construction of a nucleic acid encoding a polypeptide of interest.

b. Host Cells

The invention further provides host cells comprising a vector comprising a subject nucleic acid. Suitable host cells include prokaryotic, e.g., bacterial cells (for example E. coli ), as well as eukaryotic cells e.g. an animal cell (for example an insect, mammal, fish, amphibian, bird or reptile cell), a plant cell (for example a maize or Arabidopsis cell), or a fungal cell (for example a S. cerevisiae cell). In certain embodiments, any cell suitable for expression of a polypeptide of interest-encoding nucleic acid may be used as a host cell. Usually, an animal host cell line is used, examples of which are as follows: monkey kidney cells (COS cells), monkey kidney CVI cells transformed by SV40 (COS-7, ATCC CRL 165 1); human embryonic kidney cells (HEK-293 [“293”], Graham et al. J. Gen Virol. 36:59 (1977)); HEK-293T [“293T”] cells; baby hamster kidney cells (BHK, ATCC CCL 10); chinese hamster ovary-cells (CHO, Urlaub and Chasin, Proc. Natl. Acad. Sci. (USA) 77:4216, (1980); Syrian golden hamster cells MCB3901 (ATCC CRL-9595); mouse sertoli cells (TM4, Mather, Biol. Reprod. 23:243-251 (1980)); monkey kidney cells (CVI ATCC CCL 70); african green monkey kidney cells (VERO-76, ATCC CRL-1587); human cervical carcinoma cells (HELA, ATCC CCL 2); canine kidney cells (MDCK, ATCC CCL 34); buffalo rat liver cells (BRL 3A, ATCC CRL 1442); human lung cells (W138, ATCC CCL 75); human liver cells (hep G2, HB 8065); mouse mammary tumor (MMT 060562, ATCC CCL 51); TRI cells (Mather et al., Annals N.Y. Acad. Sci 383:44-68 (1982)); NIH/3T3 cells (ATCC CRL-1658); and mouse L cells (ATCC CCL-1). In certain embodiments, melanophores are used. Melanophores are skin cells found in lower vertebrates. Relevant materials and methods will be followed according to the disclosure of U.S. Pat. No. 5,462,856 and U.S. Pat. No. 6,051,386. These patent disclosures are hereby incorporated by reference in their entirety. Additional cell lines will become apparent to those of ordinary skill in the art, and a wide variety of cell lines are available from the American Type Culture Collection, 10801 University Boulevard, Manassas, Va. 20110-2209.

C. Screening of Candidate Compounds

1. Generic GPCR Screening Assay Techniques

When a G protein receptor becomes active, it binds to a G protein (e.g., Gq, Gs, Gi, Gz, Go) and stimulates the binding of GTP to the G protein. The G protein then acts as a GTPase and slowly hydrolyzes the GTP to GDP, whereby the receptor, under normal conditions, becomes deactivated. However, activated receptors continue to exchange GDP to GTP. A non-hydrolyzable analog of GTP, [ 35 S]GTPγS, can be used to monitor enhanced binding to membranes which express activated receptors. It is reported that [ 35 S]GTPγS can be used to monitor G protein coupling to membranes in the absence and presence of ligand. An example of this monitoring, among other examples well-known and available to those in the art, was reported by Traynor and Nahorski in 1995. The preferred use of this assay system is for initial screening of candidate compounds because the system is generically applicable to all G protein-coupled receptors regardless of the particular G protein that interacts with the intracellular domain of the receptor.

2. Specific GPCR Screening Assay Techniques

Once candidate compounds are identified using the “generic” G protein-coupled receptor assay (i.e., an assay to select compounds that are agonists or inverse agonists), in some embodiments further screening to confirm that the compounds have interacted at the receptor site is preferred. For example, a compound identified by the “generic” assay may not bind to the receptor, but may instead merely “uncouple” the G protein from the intracellular domain.

a. Gs, Gz and Gi.

Gs stimulates the enzyme adenylyl cyclase. Gi (and Gz and Go), on the other hand, inhibit adenylyl cyclase. Adenylyl cyclase catalyzes the conversion of ATP to cAMP; thus, activated GPCRs that couple the Gs protein are associated with increased cellular levels of cAMP. On the other hand, activated GPCRs that couple Gi (or Gz, Go) protein are associated with decreased cellular levels of cAMP. See, generally, “Indirect Mechanisms of Synaptic Transmission,” Chpt. 8 , From Neuron To Brain (3 rd Ed.) Nichols, J. G. et al eds. Sinauer Associates, Inc. (1992). Thus, assays that detect cAMP can be utilized to determine if a candidate compound is, e.g. an inverse agonist to the receptor (i.e., such a compound would decrease the levels of cAMP). A variety of approaches known in the art for measuring cAMP can be utilized; in some embodiments a preferred approach relies upon the use of anti-cAMP antibodies in an ELISA-based format. Another type of assay that can be utilized is a whole cell second messenger reporter system assay. Promoters on genes drive the expression of the proteins that a particular gene encodes. Cyclic AMP drives gene expression by promoting the binding of a cAMP-responsive DNA binding protein or transcription factor (CREB) that then binds to the promoter at specific sites called cAMP response elements and drives the expression of the gene. Reporter systems can be constructed which have a promoter containing multiple cAMP response elements before the reporter gene, e.g., β-galactosidase or luciferase. Thus, an activated Gs-linked receptor causes the accumulation of cAMP that then activates the gene and expression of the reporter protein. The reporter protein such as β-galactosidase or luciferase can then be detected using standard biochemical assays (Chen et al. 1995).

b. Go and Gq.

Gq and Go are associated with activation of the enzyme phospholipase C, which in turn hydrolyzes the phospholipid PIP 2 , releasing two intracellular messengers: diacycloglycerol (DAG) and inistol 1,4,5-triphoisphate (IP 3 ). Increased accumulation of IP 3 is associated with activation of Gq- and Go-associated receptors. See, generally, “Indirect Mechanisms of Synaptic Transmission,” Chpt. 8 , From Neuron To Brain (3 rd Ed.) Nichols, J. G. et al eds. Sinauer Associates, Inc. (1992). Assays that detect IP 3 accumulation can be utilized to determine if a candidate compound is, e.g., an inverse agonist to a Gq- or Go-associated receptor (i.e., such a compound would decrease the levels of IP 3 ). Gq-associated receptors can also been examined using an AP1 reporter assay in that Gq-dependent phospholipase C causes activation of genes containing AP1 elements; thus, activated Gq-associated receptors will evidence an increase in the expression of such genes, whereby inverse agonists thereto will evidence a decrease in such expression, and agonists will evidence an increase in such expression. Commercially available assays for such detection are available.

3. GPCR Fusion Protein

The use of an endogenous, constitutively active GPCR or a non-endogenous, constitutively activated GPCR, for use in screening of candidate compounds for the direct identification of inverse agonists or agonists provides an interesting screening challenge in that, by definition, the receptor is active even in the absence of an endogenous ligand bound thereto. Thus, in order to differentiate between, e.g., the non-endogenous receptor in the presence of a candidate compound and the non-endogenous receptor in the absence of that compound, with an aim of such a differentiation to allow for an understanding as to whether such compound may be an inverse agonist or agonist or have no affect on such a receptor, in some embodiments it is preferred that an approach be utilized that can enhance such differentiation. In some embodiments, a preferred approach is the use of a GPCR Fusion Protein.

Generally, once it is determined that a non-endogenous GPCR has been constitutively activated using the assay techniques set forth above (as well as others known to the art-skilled), it is possible to determine the predominant G protein that couples with the endogenous GPCR. Coupling of the G protein to the GPCR provides a signaling pathway that can be assessed. In some embodiments it is preferred that screening take place using a mammalian expression system, as such a system will be expected to have endogenous G protein therein. Thus, by definition, in such a system, the non-endogenous, constitutively activated GPCR will continuously signal. In some embodiments it is preferred that this signal be enhanced such that in the presence of, e.g., an inverse agonist to the receptor, it is more likely that it will be able to more readily differentiate, particularly in the context of screening, between the receptor when it is contacted with the inverse agonist.

The GPCR Fusion Protein is intended to enhance the efficacy of G protein coupling with the non-endogenous GPCR. The GPCR Fusion Protein is preferred for screening with either an endogenous, constitutively active GPCR or a non-endogenous, constitutively activated GPCR because such an approach increases the signal that is generated in such screening techniques. This is important in facilitating a significant “signal to noise” ratio; such a significant ratio is preferred for the screening of candidate compounds as disclosed herein.

The construction of a construct useful for expression of a GPCR Fusion Protein is within the purview of those having ordinary skill in the art. Commercially available expression vectors and systems offer a variety of approaches that can fit the particular needs of an investigator. Important criteria in the construction of such a GPCR Fusion Protein construct include but are not limited to, that the GPCR sequence and the G protein sequence both be in-frame (preferably, the sequence for the endogenous GPCR is upstream of the G protein sequence), and that the “stop” codon of the GPCR be deleted or replaced such that upon expression of the GPCR, the G protein can also be expressed. The GPCR can be linked directly to the G protein, or there can be spacer residues between the two (preferably, no more than about 12, although this number can be readily ascertained by one of ordinary skill in the art). Based upon convenience, it is preferred to use a spacer. In some embodiments it is preferred, that the G protein that couples to the non-endogenous GPCR will have been identified prior to the creation of the GPCR Fusion Protein construct. Because there are only a few G proteins that have been identified, it is preferred that a construct comprising the sequence of the G protein (i.e., a universal G protein construct, see Example 5(a) below) be available for insertion of an endogenous GPCR sequence therein; this provides for further efficiency in the context of large-scale screening of a variety of different endogenous GPCRs having different sequences.

As noted above, activated GPCRs that couple to Gi, Gz and Go are expected to inhibit the formation of cAMP making assays based upon these types of GPCRs challenging [i.e., the cAMP signal decreases upon activation, thus making the direct identification of, e.g., agonists (which would further decrease this signal) challenging]. As will be disclosed herein, it has been ascertained that for these types of receptors, it is possible to create a GPCR Fusion Protein that is not based upon the GPCR's endogenous G protein, in an effort to establish a viable cyclase-based assay. Thus, for example, an endogenous Gi coupled receptor can be fused to a Gs protein—such a fusion construct, upon expression, “drives” or “forces” the endogenous GPCR to couple with, e.g., Gs rather than the “natural” Gi protein, such that a cyclase-based assay can be established. Thus, for Gi, Gz and Go coupled receptors, in some embodiments it is preferred that when a GPCR Fusion Protein is used and the assay is based upon detection of adenylyl cyclase activity, that the fusion construct be established with Gs (or an equivalent G protein that stimulates the formation of the enzyme adenylyl cyclase).

Equally effective is a G Protein Fusion construct that utilizes a Gq Protein fused with a Gs, Gi, Gz or Go Protein. In some embodiments a preferred fusion construct can be accomplished with a Gq Protein wherein the first six (6) amino acids of the G-protein α-subunit (“Gαq”) is deleted and the last five (5) amino acids at the C-terminal end of Gαq is replaced with the corresponding amino acids of the Gα of the G protein of interest. For example, a fusion construct can have a Gq (6 amino acid deletion) fused with a Gi Protein, resulting in a “Gq/Gi Fusion Construct”. This fusion construct will forces the endogenous Gi coupled receptor to couple to its non-endogenous G protein, Gq, such that the second messenger, for example, inositol triphosphate or diacylgycerol, can be measured in lieu of cAMP production.

4. Co-Transfection of a Target Gi Coupled GPCR with a Signal-Enhancer Gs Coupled GPCR (cAMP Based Assays)

A Gi coupled receptor is known to inhibit adenylyl cyclase, and, therefore, decreases the level of cAMP production, which can make the assessment of cAMP levels challenging. In certain embodiments, an effective technique in measuring the decrease in production of cAMP as an indication of activation of a receptor that predominantly couples Gi upon activation can be accomplished by co-transfecting a signal enhancer, e.g., a non-endogenous, constitutively activated receptor that predominantly couples with Gs upon activation (e.g. TSHR-A623I; see infra), with the Gi linked GPCR. As is apparent, activation of a Gs coupled receptor can be determined based upon an increase in production of cAMP. Activation of a Gi coupled receptor leads to a decrease in production cAMP. Thus, the co-transfection approach is intended to advantageously exploit these “opposite” affects. For example, co-transfection of a non-endogenous, constitutively activated Gs coupled receptor (the “signal enhancer”) with expression vector alone provides a baseline cAMP signal (i.e., although the Gi coupled receptor will decrease cAMP levels, this “decrease” will be relative to the substantial increase in cAMP levels established by constitutively activated Gs coupled signal enhancer). By then co-transfecting the signal enhancer with the “target receptor”, an inverse agonist of the Gi coupled target receptor will increase the measured cAMP signal, while an agonist of the Gi coupled target receptor will decrease this signal.

Candidate compounds that are directly identified using this approach should be assessed independently to ensure that these do not target the signal enhancing receptor (this can be done prior to or after screening against the co-transfected receptors).

D. Medicinal Chemistry

Candidate Compounds

Any molecule known in the art can be tested for its ability to modulate (increase or decrease) the activity of a GPCR of the present invention. For identifying a compound that modulates activity, candidate compounds can be directly provided to a cell expressing the receptor.

This embodiment of the invention is well suited to screen chemical libraries for molecules which modulate, e.g., inhibit, antagonize, or agonize, the amount of, or activity of, a receptor. The chemical libraries can be peptide libraries, peptidomimetic libraries, chemically synthesized libraries, recombinant, e.g., phage display libraries, and in vitro translation-based libraries, other non-peptide synthetic organic libraries, etc. This embodiment of the invention is also well suited to screen endogenous candidate compounds comprising biological materials, including but not limited to plasma and tissue extracts, and to screen libraries of endogenous compounds known to have biological activity.

In some embodiments direct identification of candidate compounds is conducted in conjunction with compounds generated via combinatorial chemistry techniques, whereby thousands of compounds are randomly prepared for such analysis. The candidate compound may be a member of a chemical library. This may comprise any convenient number of individual members, for example tens to hundreds to thousand to millions of suitable compounds, for example peptides, peptoids and other oligomeric compounds (cyclic or linear), and template-based smaller molecules, for example benzodiazepines, hydantoins, biaryls, carbocyclic and polycyclic compounds (e.g., naphthalenes, phenothiazines, acridines, steroids etc.), carbohydrate and amino acid derivatives, dihydropyridines, benzhydryls and heterocycles (e.g., trizines, indoles, thiazolidines etc.). The numbers quoted and the types of compounds listed are illustrative, but not limiting. Preferred chemical libraries comprise chemical compounds of low molecular weight and potential therapeutic agents.

Exemplary chemical libraries are commercially available from several sources (ArQule, Tripos/PanLabs, ChemDesign, Pharmacopoeia). In some cases, these chemical libraries are generated using combinatorial strategies that encode the identity of each member of the library on a substrate to which the member compound is attached, thus allowing direct and immediate identification of a molecule that is an effective modulator. Thus, in many combinatorial approaches, the position on a plate of a compound specifies that compound's composition. Also, in one example, a single plate position may have from 1-20 chemicals that can be screened by administration to a well containing the interactions of interest. Thus, if modulation is detected, smaller and smaller pools of interacting pairs can be assayed for the modulation activity. By such methods, many candidate molecules can be screened.

Many diversity libraries suitable for use are known in the art and can be used to provide compounds to be tested according to the present invention. Alternatively, libraries can be constructed using standard methods. Further, more general, structurally constrained, organic diversity (e.g., nonpeptide) libraries, can also be used. By way of example, a benzodiazepine library (see e.g., Bunin et al., 1994, Proc. Natl. Acad. Sci. USA 91:4708-4712) may be used.

In another embodiment of the present invention, combinatorial chemistry can be used to identify modulators of the GPCRs of the present invention. Combinatorial chemistry is capable of creating libraries containing hundreds of thousands of compounds, many of which may be structurally similar. While high throughput screening programs are capable of screening these vast libraries for affinity for known targets, new approaches have been developed that achieve libraries of smaller dimension but which provide maximum chemical diversity. (See e.g., Matter, 1997, Journal of Medicinal Chemistry 40:1219-1229).

One method of combinatorial chemistry, affinity fingerprinting, has previously been used to test a discrete library of small molecules for binding affinities for a defined panel of proteins. The fingerprints obtained by the screen are used to predict the affinity of the individual library members for other proteins or receptors of interest (in the instant invention, the receptors of the present invention). The fingerprints are compared with fingerprints obtained from other compounds known to react with the protein of interest to predict whether the library compound might similarly react. For example, rather than testing every ligand in a large library for interaction with a complex or protein component, only those ligands having a fingerprint similar to other compounds known to have that activity could be tested. (See, e.g., Kauvar et al., 1995, Chemistry and Biology 2:107-118; Kauvar, 1995, Affinity fingerprinting, Pharmaceutical Manufacturing International. 8:25-28; and Kauvar, Toxic-Chemical Detection by Pattern Recognition in New Frontiers in Agrochemical Immunoassay, D. Kurtz. L. Stanker and J. H. Skerritt. Editors, 1995, AOAC: Washing