Cytokine receptor modulators, method of identifying same, and method of modulating cytokine receptors activity with same

Abstract

The present invention relates to a method for identifying a non-competitive peptide, which inhibits the activity of a cytokine receptor. This method includes the steps of selecting a candidate peptide containing from about 7 to about 20 amino acids derived from a flexible region of a cytokine receptor, and determining the ability of the peptide to inhibit or promote the oligomerization and/or activation of the receptor by measuring an activity of the receptor in the absence or the presence of the candidate peptide, wherein the non-competitive peptide is selected when the activity of the receptor is measurably lower in the presence of the peptide as compared to in the absence of the peptide so identified. This invention also provides agonists of cytokine receptor activity. Pharmaceutical compositions that comprise the identified peptides are disclosed. Also disclosed are methods for treating patients with a disease or condition associated with abnormal cytokine receptor mediated function or activity such as inflammatory, autoimmune and vascular diseases.

1 - 37 . (canceled) 
     
     
         38 . A method of identifying a peptide modulator of a receptor comprising steps of:
 (a) identifying an extracellular region that is involved in conformational change or oligomerization of a receptor;   (b) providing a candidate peptide that is up to 20 amino acids long and has an amino acid sequence corresponding to at least 5 contiguous amino acids that appear in the extracellular region identified in step (a); and   (c) determining if the candidate peptide inhibits or activates an activity of the receptor relative to a control.   
     
     
         39 . The method of  claim 38 , wherein the extracellular region involved in conformational change or oligomerization of the receptor is selected from the group consisting of juxtamembranous regions, regions containing α helix, β sheet, loops and/or β turns, regions between domains, regions between two β chains, and combinations thereof. 
     
     
         40 . The method of  claim 38 , wherein the extracellular region involved in conformational change or oligomerization of the receptor is identified by structure modeling. 
     
     
         41 . The method of  claim 38 , wherein the extracellular region involved in conformational change or oligomerization of the receptor is identified by sequence alignment. 
     
     
         42 . The method of  claim 38 , wherein the candidate peptide has an amino acid sequence identical to 5-20 contiguous amino acids that appear in the extracellular region identified in step (a). 
     
     
         43 . The method of  claim 38 , wherein the candidate peptide has an amino acid sequence otherwise identical to 5-20 contiguous amino acids that appear in the extracellular region identified in step (a), but incorporating one or more D-amino acid substitutions for corresponding L-amino acids. 
     
     
         44 . The method of  claim 38 , wherein the candidate peptide comprises one or more modifications to increase protease resistance, serum stability and/or bioavailability. 
     
     
         45 . The method of  claim 44 , wherein the one or more modifications are selected from N- and/or C-terminal acetylation, glycosylation, biotinylation, D-amino acid, un-natural amino acid and/or cyclic amino acid substitution. 
     
     
         46 . The method of  claim 38 , wherein the candidate peptide comprises additional amino acids that do not appear within the 5-20 contiguous amino acids in the extracellular region identified in step (a). 
     
     
         47 . The method of  claim 46 , wherein the additional amino acids are from a separate region of the receptor. 
     
     
         48 . The method of  claim 38 , wherein the control indicates the activity of the receptor in the absence of the candidate peptide. 
     
     
         49 . The method of  claim 38 , further comprising a step of identifying an peptide inhibitor of the receptor based on the result from step (c). 
     
     
         50 . The method of  claim 38 , wherein the receptor is a cytokine receptor, a growth factor receptor, or a chemokine receptor. 
     
     
         51 . The method of  claim 38 , wherein the receptor is a cytokine receptor selected from the group consisting of VEGF receptors, PDGF receptors, IGF-1 receptors, FGF receptors, EGF receptors, interleukin receptors, IFNα receptor, IFNβ receptor, TGFβ receptor, NGF/TNF receptors and combinations thereof. 
     
     
         52 . The method of  claim 51 , wherein the receptor is an interleukin type I and/or type II receptor for interleukins 1, 2, 3, 4, 5, 7, 9 or 15. 
     
     
         53 . The method of  claim 51 , wherein step (c) comprises performing an in vitro cytokine receptor activity assay. 
     
     
         54 . The method of  claim 53 , wherein the in vitro cytokine receptor activity assay is selected from a cellular proliferation assay, a kinase assay, PGE2 synthesis assay, IL-6 synthesis assay, collagenase expression assay, Akt assay, and/or VCAM-1 expression assay. 
     
     
         55 . The method of  claim 53 , wherein the control indicates the activity of the cytokine receptor measured by the in vitro assay in the absence of the candidate peptide. 
     
     
         56 . The method of  claim 55 , further comprising a step of identifying an peptide inhibitor of the cytokine receptor based on the result from step (c). 
     
     
         57 . The method of  claim 56 , wherein the method further comprises a step of testing the ability of the identified peptide inhibitor to treat a disease, disorder, or condition associated with deregulation or malfunction of the cytokine receptor. 
     
     
         58 . The method of  claim 57 , wherein the testing step comprises treating an animal model of the disease, disorder, or condition associated with deregulation or malfunction of the cytokine receptor using the peptide inhibitor. 
     
     
         59 . A peptide modulator identified using the method of  claim 38 . 
     
     
         60 . A peptide inhibitor identified using the method of  claim 49 . 
     
     
         61 . A method of inhibiting an activity of a receptor comprising
 contacting the receptor with a peptide that is up to 20 amino acids long and has an amino acid sequence corresponding to at least 5 contiguous amino acids that appear in an extracellular region of the receptor,   wherein the extracellular region is selected from the group consisting of juxtamembranous regions, regions containing α helix, β sheet, loops and/or β turns, regions between domains, regions between two β chains, and combinations thereof.   
     
     
         62 . The method of  claim 61 , wherein the peptide has an amino acid sequence identical to 5-20 contiguous amino acids that appear in the extracellular region of the receptor. 
     
     
         63 . The method of  claim 61 , wherein the peptide has an amino acid sequence otherwise identical to 5-20 contiguous amino acids that appear in the extracellular region of the receptor, but incorporating one or more D-amino acid substitutions for corresponding L-amino acids. 
     
     
         64 . The method of  claim 61 , wherein the peptide comprises one or more modifications to increase protease resistance, serum stability and/or bioavailability. 
     
     
         65 . The method of  claim 64 , wherein the one or more modifications are selected from N- and/or C-terminal acetylation, glycosylation, biotinylation, D-amino acid, un-natural amino acid and/or cyclic amino acid substitution. 
     
     
         66 . The method of  claim 61 , wherein the receptor is a cytokine receptor, a growth factor receptor, or a chemokine receptor. 
     
     
         67 . The method of  claim 61 , wherein the receptor is a cytokine receptor selected from the group consisting of VEGF receptors, PDGF receptors, IGF-1 receptors, FGF receptors, EGF receptors, interleukin receptors, IFNα receptor, IFNβ receptor, TGFβ receptor, NGF/TNF receptors and combinations thereof. 
     
     
         68 . The method of  claim 67 , wherein the receptor is an interleukin type I and/or type II receptor for interleukins 1, 2, 3, 4, 5, 7, 9 or 15.