Reagents and Methods for Smooth Muscle Therapies

Abstract

The present invention provides novel polypeptides comprising heat shock protein 20 (HSP20)-derived polypeptides to treat or inhibit smooth muscle vasospasm, as well to treat and inhibit smooth muscle cell proliferation and migration.

1 . A polypeptide consisting of a sequence according to general formula I:
   X1-X2-[X3-A(X4)APLP-X5-] u -X6   wherein X1 is absent or is one or more molecules comprising one or more aromatic ring;   X2 is absent or comprises a transduction domain;   X3 is 0, 1, 2, 3, or 4 amino acids of the sequence WLRR (SEQ ID NO:1);   X4 is selected from the group consisting of S, T, Y, D, E, phosphoserine analogs and phosphotyrosine analogs;   X5 is 0, 1, 2, or 3 amino acids of a sequence of genus Z1-Z2-Z3,   wherein Z1 is selected from the group consisting of G and D;   Z2 is selected from the group consisting of L and K; and   Z3 is selected from the group consisting of S and T; and   X6 is absent or comprises a transduction domain; and   wherein u is 1-5.   
     
     
         2 . The polypeptide of  claim 1  wherein either X2 or X6 comprises a transduction domain. 
     
     
         3 . The polypeptide of  claim 1  wherein X4 is phosphorylated. 
     
     
         4 . The polypeptide of  claim 1  wherein X1 is a molecule comprising an aromatic ring. 
     
     
         5 . The polypeptide of  claim 4  wherein X1 is selected from the group consisting of F, Y, W; and compounds comprising 9-fluoroenylmethyl. 
     
     
         6 . A polypeptide comprising a sequence according to general formula II:
   X1-X2-[X3-A(X4)APLP-X5] u -X6   wherein X1 is absent or is one or more molecules comprising one or more aromatic ring;   X2 is absent or comprises a cell transduction domain;   X3 is 0-14 amino acids of the sequence of heat shock protein 20 between residues 1 and 14 of SEQ ID NO:297;   X4 is selected from the group consisting of S, T, Y, D, E, phosphoserine analogs and phosphotyrosine analogs;   X5 is 0-140 amino acids of heat shock protein 20 between residues 21 and 160 of SEQ ID NO:297;   X6 is absent or comprises a cell transduction domain; and   wherein at least one of X2 and X6 comprise a transduction domain.   
     
     
         6 . The polypeptide of  claim 5  wherein X4 is phosphorylated. 
     
     
         7 . The polypeptide of  claim 5  wherein X1 is a molecule comprising an aromatic ring. 
     
     
         8 . The polypeptide of  claim 7  wherein X1 is selected from the group consisting of F, Y, W; and compounds comprising 9-fluoroenylmethyl. 
     
     
         9 . A pharmaceutical composition, comprising one or more polypeptides according to  claim 1 , and a pharmaceutically acceptable carrier. 
     
     
         10 . A pharmaceutical composition, comprising one or more polypeptides according to  claim 5 , and a pharmaceutically acceptable carrier. 
     
     
         11 . An isolated nucleic acid sequence encoding the polypeptide of  claim 1 . 
     
     
         12 . An isolated nucleic acid sequence encoding the polypeptide of  claim 5 . 
     
     
         13 . An expression vector comprising the nucleic acid of  claim 11 . 
     
     
         14 . An expression vector comprising the nucleic acid of  claim 12 . 
     
     
         15 . A host cell comprising the expression vector of  claim 13 . 
     
     
         16 . A host cell comprising the expression vector of  claim 14 . 
     
     
         17 . An improved biomedical device, wherein the biomedical device comprises one or more polypeptides according to  claim 1  disposed on or in the biomedical device. 
     
     
         18 . An improved biomedical device, wherein the biomedical device comprises one or more polypeptides according to  claim 5  disposed on or in the biomedical device. 
     
     
         19 . A method for inhibiting smooth muscle cell proliferation and/or migration, comprising contacting the smooth muscle cells with an amount effective to inhibit smooth muscle cell proliferation and/or migration of one or more polypeptides according to  claim 1 . 
     
     
         20 . A method for inhibiting smooth muscle cell proliferation and/or migration, comprising contacting the smooth muscle cells with an amount effective to inhibit smooth muscle cell proliferation and/or migration of one or more polypeptides according to  claim 5 . 
     
     
         21 . A method for inhibiting smooth muscle cell proliferation and/or migration, comprising contacting the smooth muscle cells with an amount effective to inhibit smooth muscle cell proliferation and/or migration of HSP20, or a functional equivalent thereof. 
     
     
         22 . A method for treating or inhibiting a disorder selected from the group consisting of intimal hyperplasia, stenosis, restenosis, and/or atherosclerosis, comprising contacting a subject in need thereof with an amount effective to treat or inhibit intimal hyperplasia, stenosis, restenosis, and/or atherosclerosis of one or more polypeptides according to  claim 1 . 
     
     
         23 . A method for treating or inhibiting a disorder selected from the group consisting of intimal hyperplasia, stenosis, restenosis, and/or atherosclerosis, comprising contacting a subject in need thereof with an amount effective to treat or inhibit intimal hyperplasia, stenosis, restenosis, and/or atherosclerosis of one or more polypeptides according to  claim 5 . 
     
     
         24 . A method for treating or inhibiting a disorder selected from the group consisting of intimal hyperplasia, stenosis, restenosis, and/or atherosclerosis, comprising contacting a patient in need thereof with an amount effective to treat or inhibit intimal hyperplasia, stenosis, restenosis, and/or atherosclerosis of HSP20, or a functional equivalent thereof. 
     
     
         25 . A method for treating or inhibiting smooth muscle spasm, comprising contacting a subject in need thereof with an amount effective to inhibit vasoconstriction of one or more polypeptides according to  claim 1 . 
     
     
         26 . A method for treating or inhibiting smooth muscle spasm, comprising contacting a subject in need thereof with an amount effective to inhibit vasoconstriction of one or more polypeptides according to  claim 5 . 
     
     
         27 . A method for treating or inhibiting smooth muscle spasm, comprising contacting a subject in need thereof with an amount effective to inhibit vasoconstriction of HSP20, or a functional equivalent thereof. 
     
     
         28 . A composition comprising:
 (a) a polypeptide according to  claim 1 ; and   (b) an inhibitor of HSP27.   
     
     
         29 . A composition comprising:
 (a) a polypeptide according to  claim 5 ; and   (b) an inhibitor of HSP27.   
     
     
         30 . A method for inhibiting smooth muscle spasm, comprising contacting a graft with an amount effective to inhibit vasoconstriction of one or more polypeptides of  claim 1 . 
     
     
         31 . A method for inhibiting smooth muscle spasm, comprising contacting a graft with an amount effective to inhibit vasoconstriction of one or more polypeptides of  claim 5 . 
     
     
         32 . A method for inhibiting smooth muscle spasm, comprising contacting a graft with an amount effective to inhibit vasoconstriction of HSP20, or a functional equivalent thereof.