Regulation of function of angiopoietin

Abstract

Pharmaceutical compositions that comprise a pharmaceutically acceptable carrier and either a therapeutically effective amount of an ECM-binding fragment of Ang-1 protein that comprises SEQ ID NO:1 and/or SEQ ID NO:2 or a homologous peptide thereof and pharmaceutical compositions that comprise a pharmaceutically acceptable carrier and a vector comprising a nucleic acid molecule that comprises the nucleotide sequence that encodes an ECM-binding fragment of Ang-1 protein that comprises SEQ ID NO:1 and/or SEQ ID NO:2 or a homologous peptide thereof are disclosed. Methods of using such compositions to treat individuals suspected of having coronary artery disease, vascular disease or a condition involving ischemia and to promote angiogenesis, endothelial survival and maintaining vascular integrity are disclosed. Methods to identify compounds that modulates binding of Ang-1 to ECM are disclosed. Pharmaceutical compositions which comprise a therapeutically effective amount of Ang-2 protein and/or a vector comprising a nucleic acid molecule that comprises the nucleotide coding sequence of Ang-2 and methods of using such compositions to treat individuals suspected of having cancer are disclosed.

1 . A pharmaceutical composition comprising
 a) a pharmaceutically acceptable carrier and   b) a therapeutically effective amount of a vector comprising a nucleic acid molecule that comprises the nucleotide sequence that encodes an ECM-binding fragment of Ang-1 protein that comprises SEQ ID NO:1 and/or SEQ ID NO:2 or a homologous peptide thereof.   
     
     
         2 . The pharmaceutical composition of  claim 1  further comprising a therapeutically effective amount of an ECM-binding fragment of Ang-1 protein that comprises SEQ ID NO:1 and/or SEQ ID NO:2 or a homologous peptide thereof. 
     
     
         3 . The pharmaceutical composition of  claim 1  comprising a vector comprising a nucleic acid molecule that comprises the nucleotide sequence that encodes an ECM-binding fragment of Ang-1 protein that comprises SEQ ID NO:1 and/or SEQ ID NO:2. 
     
     
         4 . The pharmaceutical composition of  claim 3  comprising wherein said vector is a viral vector. 
     
     
         5 . The pharmaceutical composition of  claim 3  comprising wherein said vector is a DNA plasmid. 
     
     
         6 . A method of treating an individual suspected of having coronary artery disease, vascular disease or a condition involving ischemia comprising the step of administering to said individual a pharmaceutical composition comprising
 a) a pharmaceutically acceptable carrier and   b) therapeutically effective amount of a vector comprising a nucleic acid molecule that comprises the nucleotide sequence that encodes an ECM-binding fragment of Ang-1 protein that comprises SEQ ID NO:1 and/or SEQ ID NO:2 or a homologous peptide thereof.   
     
     
         7 . The method of  claim 6  wherein said pharmaceutical composition further comprises a therapeutically effective amount of an ECM-binding fragment of Ang-1 protein that comprises SEQ ID NO:1 and/or SEQ ID NO:2 or a homologous peptide thereof. 
     
     
         8 . The method of  claim 6  wherein said pharmaceutical composition comprises a vector comprising a nucleic acid molecule that comprises the nucleotide sequence that encodes an ECM-binding fragment of Ang-1 protein that comprises SEQ ID NO:1 and/or SEQ ID NO:2. 
     
     
         9 . A method of promoting angiogenesis, endothelial survival and maintaining vascular integrity in an individual comprising the step of administering to said individual a pharmaceutical composition comprising
 a) a pharmaceutically acceptable carrier and   b) therapeutically effective amount of a vector comprising a nucleic acid molecule that comprises the nucleotide sequence that encodes an ECM-binding fragment of Ang-1 protein that comprises SEQ ID NO:1 and/or SEQ ID NO:2 or a homologous peptide thereof.   
     
     
         10 . The method of  claim 9  wherein said pharmaceutical composition further comprises a therapeutically effective amount of an ECM-binding fragment of Ang-1 protein that comprises SEQ ID NO:1 and/or SEQ ID NO:2 or a homologous peptide thereof. 
     
     
         11 . The method of  claim 9  wherein said pharmaceutical composition comprises a vector comprising a nucleic acid molecule that comprises the nucleotide sequence that encodes an ECM-binding fragment of Ang-1 protein that comprises SEQ ID NO:1 and/or SEQ ID NO:2. 
     
     
         12 . A method of identifying compounds that modulates binding of Ang-1 to ECM comprising performing a test assay that comprises the steps of:
 a) contacting a protein that comprises at least an ECM-binding fragment of Ang-1 protein that comprises SEQ ID NO:1 and/or SEQ ID NO:2 with ECM material in the presence of a test compound;   b) measuring the level of binding of said protein that comprises at least an ECM-binding fragment of Ang-1 protein that comprises SEQ ID NO:1 and/or SEQ ID NO:2 with said ECM; and   c) comparing said level with the level of binding of protein that comprises at least an ECM-binding fragment of Ang-1 protein that comprises SEQ ID NO: and/or SEQ ID NO:2 with ECM material in the absence of said test compound;   wherein   when the level of binding of said protein that comprises at least an ECM-binding fragment of Ang-1 protein that comprises SEQ ID NO:1 and/or SEQ ID NO:2 with said ECM in the presence of said test compound is less than the level of binding of said protein that comprises at least an ECM-binding fragment of Ang-1 protein that comprises SEQ ID NO:1 and/or SEQ ID NO:2 with said ECM in the absence of said test compound results indicate that the test compound modulates binding of Ang-1 to ECM by inhibiting said binding and   when the level of binding of said protein that comprises at least an ECM-binding fragment of Ang-1 protein that comprises SEQ ID NO:1 and/or SEQ ID NO:2 with said ECM in the presence of said test compound is more than the level of binding of said protein that comprises at least an ECM-binding fragment of Ang-1 protein that comprises SEQ ID NO:1 and/or SEQ ID NO:2 with said ECM in the absence of said test compound results indicate that the test compound modulates binding of Ang-1 to ECM by enhancing said binding.   
     
     
         13 . The method of  claim 12  wherein the protein is Ang-1 protein. 
     
     
         14 . The method of  claim 12  wherein the protein is a fragment of Ang-1 protein. 
     
     
         15 . The method of  claim 12  wherein the protein comprises a detectable label. 
     
     
         16 . The method of  claim 15  wherein the level of binding is measured by measuring the amount of detectable label present on said ECM after removing unbound protein from said ECM material. 
     
     
         17 . The method of  claim 12  wherein the ECM material is produced by culturing cells on a substrate for a sufficient time for said cells to produce ECM material on said substrate and removing said cells from said substrate without removing said ECM material. 
     
     
         18 . The method of  claim 17  wherein said cells are Lewis lung carcinoma cells or TA3 murine mammary carcinoma cells. 
     
     
         19 . The method of  claim 12  wherein said method comprising multiple test assays which are identical except that the amount of test compound used differs. 
     
     
         20 . The method of  claim 12  the level of binding of protein that comprises at least an ECM-binding fragment of Ang-1 protein that comprises SEQ ID NO; I and/or SEQ ID NO:2 with ECM material in the absence of said test compound is determined by performing a control assay wherein said control assay comprises the steps of
 a) contacting a protein that comprises at least an ECM-binding fragment of Ang-1 protein that comprises SEQ ID NO:1 and/or SEQ ID NO:2 with ECM material in the absence of a test compound;   b) measuring the level of binding of said protein that comprises at least an ECM-binding fragment of Ang-1 protein that comprises SEQ ED NO:1 and/or SEQ ID NO:2 with said ECM.   
     
     
         21 . A pharmaceutical composition comprising
 a) a pharmaceutically acceptable carrier and   b) a therapeutically effective amount of Ang-2 protein and/or a vector comprising a nucleic acid molecule that comprises the nucleotide coding sequence of Ang-2.   
     
     
         22 . The pharmaceutical composition of  claim 21  comprising a therapeutically effective amount of Ang-2 protein. 
     
     
         23 . The pharmaceutical composition of  claim 21  comprising a vector comprising a nucleic acid molecule that comprises the nucleotide coding sequence of Ang-2. 
     
     
         24 . The pharmaceutical composition of  claim 23  comprising wherein said vector is a viral vector. 
     
     
         25 . The pharmaceutical composition of  claim 23  comprising wherein said vector is a DNA plasmid. 
     
     
         26 . A method of treating an individual suspected of having cancer comprising the step of administering to said individual a pharmaceutical composition comprising
 a) a pharmaceutically acceptable carrier and   b) a therapeutically effective amount of Ang-2 protein and/or a vector comprising a nucleic acid molecule that comprises the nucleotide coding sequence of Ang-2.   
     
     
         27 . The method of  claim 26  wherein said pharmaceutical composition comprises a therapeutically effective amount of Ang-2 protein. 
     
     
         28 . The method of  claim 26  wherein said pharmaceutical composition comprises a vector comprising a nucleic acid molecule that comprises the nucleotide coding sequence of Ang-2. 
     
     
         29 . The method of  claim 28  wherein said vector is a viral vector. 
     
     
         30 . The method of  claim 28  wherein said vector is a DNA plasmid.