Bi-functional arginine-glycine-aspartic acid (rgd) peptides and methods to promote angiogenesis

Abstract

The present invention provides an in vitro method for identifying a compound that promotes endothelial cell adhesion, endothelial cell spreading, endothelial cell migration and/or endothelial cell proliferation for the manufacture of a diagnostic or therapeutic agent. The present invention further provides the identified compounds and pharmaceutical compositions, and assays and kits for identifying a compound or using a compound that promotes endothelial cell adhesion, endothelial cell spreading, endothelial cell migration and/or endothelial cell proliferation and is useful for bioprinting.

1 - 14 . (canceled) 
     
     
         15 . A peptide that binds to an endothelial cell integrin, the peptide selected from the group consisting of a peptide having the following amino acid sequence:
 TFALRGDNP (SEQ ID NO:1), TFALRADNP (SEQ ID NO:2), DVEKRGDREEAHVP (SEQ ID NO:3), IQRGDIDAMIS (SEQ ID NO:4), DAVKQLQAAERGDA (SEQ ID NO:5), PMQKMRGDVFSP (SEQ ID NO:6), RSDGTG (SEQ ID NO:7), EAPRGDVYQG (SEQ ID NO:8), GLOGERGRO (SEQ ID NO:9), GFOGERGVQ (SEQ ID NO:10), DGEA (SEQ ID NO:11), GFOGER (SEQ ID NO:12), GLKGEN (SEQ ID NO:13), LDV (SEQ ID NO:14), REDV (SEQ ID NO:15), PEDGIHE (SEQ ID NO:16), PHSRN (SEQ ID NO:17), ALNGR (SEQ ID NO:18), IAFQRN (SEQ ID NO:19), IKLLI (SEQ ID NO:20), SIKVAV (SEQ ID NO:21), AGQWHRVSVRWG (SEQ ID NO:22), TWSQKALHHRVP (SEQ ID NO:23), SIYITRF (SEQ ID NO:24), SYWYRIEASRTG(SEQ ID NO:25) YIGSR (SEQ ID NO:26), RDIAEIIKDI (SEQ ID NO:27), VFDNFVLK(SEQ ID NO:28) , AEIDGIEL (SEQ ID NO:29), SETQRGDVFVP (SEQ ID NO:30), PASYRGDSC (SEQ ID NO:31), VTGRGDSPAS (SEQ ID NO:32), PQVTRGDVFTMP (SEQ ID NO:37), or variants at least 90% identical thereto.   
     
     
         16 . A pharmaceutical composition comprising the peptide of  claim 15  and a pharmaceutically acceptable carrier, diluent, or excipient. 
     
     
         17 . A hydrogel composition comprising
 at least one integrin-binding peptide of  claim 15 .   
     
     
         18 . A hydrogel composition comprising:
 (a) at least one integrin-binding peptide of  claim 15 ; and   (b) a biocompatible polymer, wherein the integrin-binding peptide is linked to the biocompatible polymer.   
     
     
         19 . The hydrogel composition of  claim 17 , wherein the linkage is a covalent or non-covalent linkage. 
     
     
         20 . The hydrogel composition of  claim 18 , wherein the biocompatible polymer is functionalized with a VEGF or a VEGF mimetic peptide and an integrin-binding peptide that binds at least one type of endothelial cell integrin. 
     
     
         21 . The hydrogel composition of  claim 20 , wherein the VEGF or the VEGF mimetic peptide is attached to the biocompatible polymer through a matrix metalloproteinase (MMP) degradable peptide linkage. 
     
     
         22 . The hydrogel composition of  claim 18 , wherein the biocompatible polymer is functionalized with a VEGF or a VEGF mimetic peptide and an integrin binding peptide that binds to an α v β 3  and/or VLA-6 integrin. 
     
     
         23 . The hydrogel composition of  claim 18 , wherein the biocompatible polymer comprises an agarose gel, polyethylene glycol, alginate, hyaluronic acid, polyacryylic acid, polyacrylic amide, polyvinyl alcohol, polyhydroxyethyl methacrylate, methacrylated dextrans, poly(N-isopropylacrylamide), or any combination thereof. 
     
     
         24 . The hydrogel composition of  claim 18 , wherein the biocompatible polymer is oxidized alginate. 
     
     
         25 . The hydrogel composition of  claim 18 , wherein the hydrogel composition is an injectable composition. 
     
     
         26 . A method of promoting angiogenesis in a subject in need thereof, comprising administering to the subject a peptide of  claim 15 , in an amount effective to promote angiogenesis. 
     
     
         27 . A method of promoting endothelial cell adhesion, endothelial cell spreading, endothelial cell migration and/or endothelial cell proliferation, comprising administering to the subject a peptide of  claim 15 , in an amount effective to promote endothelial cell adhesion, endothelial cell spreading, endothelial cell migration and/or endothelial cell proliferation. 
     
     
         28 . A method of treating or preventing ischemic injury in a subject in need thereof, comprising administering to the subject a peptide of  claim 15 , in an amount effective to treat or prevent ischemic injury. 
     
     
         29 . A method of promoting tissue regeneration in a subject in need thereof, comprising administering to the subject a peptide of  claim 15 , in an amount effective to promote tissue regeneration. 
     
     
         30 . A method of bioprinting, comprising administering a peptide of  claim 15  to a substrate in an amount effective to promote tissue regeneration. 
     
     
         31 . A biomaterial product for bioprinting, comprising a peptide of  claim 15 . 
     
     
         32 . A kit comprising a peptide of  claim 15  and a container suitable for delivery of the peptide, pharmaceutical composition or hydrogel composition into an administration device, with optional instructions for the use thereof. 
     
     
         33 . The kit of  claim 32 , wherein the device is a parenteral administration device. 
     
     
         34 . The kit of  claim 32 , wherein the device is an intramyocardial device. 
     
     
         35 . The kit of  claim 32 , wherein the device is a bioprinter.