Peptides blocking vascular endothelial growth factor (VEGF)-mediated angiogenesis, polynucleotides encoding said peptides and methods of use thereof

Abstract

The present invention provides peptides which can interact with VEGF and inhibit VEGF interaction with KDR or anti-VEGF antibody thereby inhibiting VEGF mediated angiogenesis or angiogeneis related diseases, polynucleotide encoding the peptides, vectors containing the polynucleotides, pharmaceutical compositions containing the peptides, and methods of inhibiting angiogenesis with the peptides.

1 . A method of screening for peptide molecules capable interacting with VEGF comprising: 
 generating a random peptide library;    contacting said library to a ligand capable of binding VEGF;    selecting the molecules from said library which bind to said ligand.    
     
     
         2 . The method of  claim 1 , wherein said ligand is KDR.  
     
     
         3 . The method of  claim 2 , wherein said ligand is expressed on a membrane of a cell.  
     
     
         4 . The method of  claim 1 , wherein said ligand is an anti-VEGF antibody.  
     
     
         5 . The method of  claim 1 , wherein random peptide library comprises peptides of seven amino acids.  
     
     
         6 . A purified peptide selected from the group consisting of SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14; wherein said peptide inhibits the binding of VEGF to KDR.  
     
     
         7 . The peptide of  claim 6 , wherein said peptide is SEQ ID NO: 11.  
     
     
         8 . An isolated polynucleotide which encodes the peptide of  claim 6 .  
     
     
         9 . A recombinant vector comprising the polynucleotide of  claim 8 .  
     
     
         10 . The vector of  claim 9 , wherein said vector is selected from the group consisting of a bacterial expression vector, a yeast expression vector and a mammalian expression vector.  
     
     
         11 . The vector of  claim 10 , wherein said mammalian expression vector is a viral vector.  
     
     
         12 . A prokaryotic cell comprising the isolated polynucleotide of  claim 8 .  
     
     
         13 . A eukaryotic cell comprising the isolated polynucleotide of  claim 8 .  
     
     
         14 . A pharmaceutical composition comprising the peptide of  claim 6  and a pharmaceutically acceptable carrier.  
     
     
         15 . A purified peptide selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7; wherein said peptide inhibits the binding of anti-VEGF antibody to VEGF.  
     
     
         16 . The peptide of  claim 15 , wherein said peptide is SEQ ID NO:1.  
     
     
         17 . An isolated polynucleotide which encodes the peptide of  claim 15 .  
     
     
         18 . A recombinant vector comprising the polynucleotide of  claim 17 .  
     
     
         19 . The vector of  claim 18 , wherein said vector is selected from the group consisting of a bacterial expression vector, a yeast expression vector and a mammalian expression vector.  
     
     
         20 . The vector of  claim 19 , wherein said mammalian expression vector is a viral vector.  
     
     
         21 . A prokaryotic cell comprising the isolated polynucleotide of  claim 17 .  
     
     
         22 . A eukaryotic cell comprising the isolated polynucleotide of  claim 17 .  
     
     
         23 . A pharmaceutical composition comprising the peptide of  claim 15  and a pharmaceutically acceptable carrier.  
     
     
         24 . A purified peptide of the sequence ATWLPPR (SEQ ID NO: 1).  
     
     
         25 . The peptide of  claim 24 , wherein said peptide is capable of interacting with VEGF.  
     
     
         26 . The peptide of  claim 24 , wherein said peptide is capable of inhibiting the interaction between VEGF and KDR.  
     
     
         27 . The peptide of  claim 24 , wherein said peptide is capable of inhibiting the proliferation of vascular endothelial cells mediated by VEGF.  
     
     
         28 . The peptide of  claim 24 , wherein said peptide is capable of inhibiting angiogensis mediated by VEGF.  
     
     
         29 . A method of inhibiting angiogenesis in a patient comprising administering to said patient an effective amount of the peptide of  claim 24  to inhibit angiogenesis.  
     
     
         30 . The method of  claim 29 , wherein said patient suffers from a disease selected from the group consisting of cancer, diabetic retinopathy, psoriasis, hamangloblastoma, and Kaposi's sarcoma.  
     
     
         31 . An isolated polynucleotide encoding the peptide of  claim 24 .  
     
     
         32 . A recombinant vector comprising the polynucleotide of  claim 31 .  
     
     
         33 . The vector of  claim 32 , wherein said vector is selected from the group consisting of a bacterial expression vector, a yeast expression vector, and a mammalian expression vector.  
     
     
         34 . The vector of  claim 33 , wherein said mammalian expression vector is a viral vector.  
     
     
         35 . A prokaryotic cell comprising the isolated polynucleotide of  claim 31 .  
     
     
         36 . A eukaryotic cell comprising the vector of  claim 31 .  
     
     
         37 . A pharmaceutical composition comprising the peptide of  claim 24  and a pharmaceutically acceptable carrier.