Novel inhibitors of vascular endothelial growth factor activity, their uses and processes for their production

Abstract

The present invention is directed to novel chimeric VEGF receptor proteins comprising amino acid sequences derived from the vascular endothelial growth factor (VEGF) receptors flt-1 and KDR, including the murine homologue to the human KDR receptor FLK-1, wherein said chimeric VEGF receptor proteins bind to VEGF and antagonize the endothelial cell proliferative and angiogenic activity thereof. The present invention is also directed to nucleic acids and expression vectors encoding these chimeric VEGF receptor proteins, host cells harboring such expression vectors, pharmaceutically acceptable compositions comprising such proteins, methods of preparing such proteins and to methods utilizing such proteins for the treatment of conditions associated with undesired vascularization.

1 - 28 . (canceled)  
     
     
         29 . A method of inhibiting vascularization or angiogenesis, comprising administering a chimeric VEGF receptor protein comprising immunoglobulin (Ig)-like domains from two or more different VEGF receptor molecules, wherein said chimeric VEGF receptor protein comprises Ig-like domain 2 of flt-1 receptor or KDR receptor.  
     
     
         30 . The method of  claim 29 , wherein the Ig-like domain 2 comprises an amino acid sequence of SEQ ID NO:49 or SEQ ID NO:50.  
     
     
         31 . The method of  claim 29 , wherein the chimeric VEGF receptor protein further comprises Ig-like domain 3 of a flt-1 receptor or a KDR receptor.  
     
     
         32 . The method of  claim 31 , wherein the Ig-like domain 3 comprises an amino acid sequence of SEQ ID NO:51 or SEQ ID NO:52.  
     
     
         33 . The method of  claim 31 , wherein the Ig-like domain 2 comprises an amino acid sequence of SEQ ID NO:49 or SEQ ID NO:50 and the Ig-like domain 3 comprises an amino acid sequence of SEQ ID NO:51 or SEQ ID NO:52.  
     
     
         34 . The method of  claim 29 , wherein the chimeric VEGF receptor protein inhibits VEGF mitogenic activity.  
     
     
         35 . The method of  claim 29 , wherein the chimeric VEGF receptor protein is fused to an immunoglobulin sequence.  
     
     
         36 . The method of  claim 29 , wherein the immunoglobulin sequence comprises an IgG heavy chain sequence.  
     
     
         37 . A method of inhibiting proliferation of an endothelial cell, comprising administering a chimeric VEGF receptor protein comprising immunoglobulin (Ig)-like domains from two or more different VEGF receptor molecules, wherein said chimeric VEGF receptor protein comprises Ig-like domain 2 of flt-1 receptor or KDR receptor.  
     
     
         38 . The method of  claim 37 , wherein the Ig-like domain 2 comprises an amino acid sequence of SEQ ID NO:49 or SEQ ID NO:50.  
     
     
         39 . The method of  claim 37 , wherein the chimeric VEGF receptor protein further comprises Ig-like domain 3 of a flt-1 receptor or a KDR receptor.  
     
     
         40 . The method of  claim 39 , wherein the Ig-like domain 3 comprises an amino acid sequence of SEQ ID NO:51 or SEQ ID NO:52.  
     
     
         41 . The method of  claim 39 , wherein the Ig-like domain 2 comprises an amino acid sequence of SEQ ID NO:49 or SEQ ID NO:50 and the Ig-like domain 3 comprises an amino acid sequence of SEQ ID NO:51 or SEQ ID NO:52.  
     
     
         42 . The method of  claim 37 , wherein the chimeric VEGF receptor protein inhibits VEGF mitogenic activity.  
     
     
         43 . The method of  claim 37 , wherein the chimeric VEGF receptor protein is fused to an immunoglobulin sequence.  
     
     
         44 . The method of  claim 43 , wherein the immunoglobulin sequence comprises an IgG heavy chain sequence.  
     
     
         45 . A method of treating a condition or disorder characterized by undesired vascularization, comprising administering a chimeric VEGF receptor protein comprising immunoglobulin (Ig)-like domains from two or more different VEGF receptor molecules, wherein said chimeric VEGF receptor protein comprises Ig-like domain 2 of flt-1 receptor or KDR receptor.  
     
     
         46 . The method of  claim 45 , wherein the condition or disorder comprises a neoplastic condition or disorder.  
     
     
         47 . The method of  claim 46 , wherein the neoplastic condition or disorder comprises breast, lung, esophagus, gastric, colon, rectum, liver, ovary, cervix, prostate, pancreas, renal, or endometrium carcinoma.  
     
     
         48 . The method of  claim 46 , wherein the neoplastic condition or disorder comprises thecoma, arrhenoblastoma, endometrial hyperplasia, endometriosis, fibrosarcoma, choriocarcinoma, head cancer, neck cancer, nasopharyngeal carcinoma, laryngeal carcinoma, hepatoblastoma, Karposi's sarcoma, melanoma, skin carcinoma, hemangioma, hemangioblastoma, retinoblastoma, astrocystoma, glioblastoma, Schwannoma, oligodendroglioma, medulloblastoma, neuroblastoma, rhabdomyosarcoma, osteogenic sarcoma, leiomyosarcomas, urinary tract carcinoma, thyroid carcinoma, Wilm's tumor, abnormal vascular proliferation associated with phakomatoses, and Meigs' syndrome.  
     
     
         49 . The method of  claim 45 , wherein the condition or disorder comprises a non-neoplastic condition or disorder.  
     
     
         50 . The method of  claim 49 , wherein the non-neoplastic condition or disorder comprises rheumatoid arthritis, psoriasis, atherosclerosis, diabetic retinopathy, retrolentral fibroplasia, neovascular glaucoma, age-related macular degeneration, thyroid hyperplasia, corneal transplantation, chronic inflammation, lung inflammation, nephrotic syndrome, preclampsia, ascites, pericardial effusion, or pleural effusion.  
     
     
         51 . The method of  claim 45 , wherein the Ig-like domain 2 comprises an amino acid sequence of SEQ ID NO:49 or SEQ ID NO:50.  
     
     
         52 . The method of  claim 45 , wherein the chimeric VEGF receptor protein further comprises Ig-like domain 3 of a flt-1 receptor or a KDR receptor.  
     
     
         53 . The method of  claim 52 , wherein the Ig-like domain 3 comprises an amino acid sequence of SEQ ID NO:51 or SEQ ID NO:52.  
     
     
         54 . The method of  claim 52 , wherein the Ig-like domain 2 comprises an amino acid sequence of SEQ ID NO:49 or SEQ ID NO:50 and the Ig-like domain 3 comprises an amino acid sequence of SEQ ID NO:51 or SEQ ID NO:52.  
     
     
         55 . The method of  claim 45 , wherein the chimeric VEGF receptor protein inhibits VEGF mitogenic activity.  
     
     
         56 . The method of  claim 45 , wherein the chimeric VEGF receptor protein is fused to an immunoglobulin sequence.  
     
     
         57 . The method of  claim 56 , wherein the immunoglobulin sequence comprises an IgG heavy chain sequence.  
     
     
         58 . A polynucleotide encoding a chimeric VEGF receptor protein comprising immunoglobulin (Ig)-like domains from two or more different VEGF receptor molecules, wherein said chimeric VEGF receptor protein comprises Ig-like domain 2 of flt-1 receptor or KDR receptor.  
     
     
         59 . The polynucleotide of  claim 58 , wherein the Ig-like domain 2 comprises an amino acid sequence of SEQ ID NO:49 or SEQ ID NO:50.  
     
     
         60 . The polynucleotide of  claim 58 , wherein the chimeric VEGF receptor protein further comprises Ig-like domain 3 of a flt-1 receptor or a KDR receptor.  
     
     
         61 . The polynucleotide of  claim 60 , wherein the Ig-like domain 3 comprises an amino acid sequence of SEQ ID NO:51 or SEQ ID NO:52.  
     
     
         62 . The polynucleotide of  claim 60 , wherein the Ig-like domain 2 comprises an amino acid sequence of SEQ ID NO:49 or SEQ ID NO:50 and the Ig-like domain 3 comprises an amino acid sequence of SEQ ID NO:51 or SEQ ID NO:52.