Antibody antagonists of ve-cadherin without adverse effects on vascular permeability

Abstract

The murine epitope sequence recognized by antibody E4B9 shares 100% homology with human VE-cadherin, so this antibody was examined to determine if it cross-reacts with human VE-cadherin. Western-blot analysis of several VE-cadherin expressing human and murine cells indicated that E4B9 indeed cross-reacts with human VE-cadherin. ( FIG. 6 ). This finding facilitates development of a “humanized” E4B9 antibody and its success in the preclinical development since its anti-tumor activity can be tested extensively in several mouse models.

1 - 22 . (canceled) 
     
     
         23 . A method of inhibiting angiogenesis associated with a neoplastic disease in a mammal which comprises:
 administering a pharmaceutical composition to a mammal having a neoplastic disease for a time and in an amount effective to inhibit angiogenesis associated with the neoplastic disease, the pharmaceutical composition comprising:
 an isolated monoclonal antibody or fragment thereof capable of specifically binding to a site on a mouse or human VE-cadherin, said site being within the about first 15 N-terminal amino acids of domain 1 of the VE-cadherin, wherein said antibody or fragment thereof is capable of inhibiting VE-cadherin mediated adherens junction formation in vitro but does not exert any significant or substantial effect on paracellular permeability in vitro, and 
 a pharmaceutically acceptable carrier or diluent. 
   
     
     
         24 . The method of  claim 23 , wherein said angiogenesis associated with a neoplastic disease is associated with a solid tumor. 
     
     
         25 . The method of  claim 24 , wherein said pharmaceutical composition is administered to said mammal for a time and in an amount effective to inhibit metastasis of the tumor. 
     
     
         26 . The method of  claim 24 , wherein said tumor is selected from the group consisting of carcinomas, gliomas, sarcomas, adenocarcinomas, adenosarcomas, adenomas, leukemic tumors and lymphoid tumors. 
     
     
         27 . The method of  claim 24 , wherein the antibody or fragment thereof does not exert any significant or substantial effect on vascular permeability in vivo. 
     
     
         28 . The method of  claim 24 , wherein the antibody or fragment thereof inhibits formation of new adherens junctions without disturbing existing adherens junctions. 
     
     
         29 . The method of  claim 24 , wherein the antibody or fragment thereof is a single chain antibody, is humanized, is chimerized or is bispecific. 
     
     
         30 . The method of  claim 24 , wherein the antibody or fragment thereof is fused to a heterologous polypeptide. 
     
     
         31 . A method of treating a neoplastic disease associated with vascularization in a mammal, which comprises:
 administering a pharmaceutical composition to a mammal having a neoplastic disease associated with vascularization in an amount effective to inhibit proliferation of endothelial cells without disturbing normal vasculature, the pharmaceutical composition comprising:
 an isolated monoclonal antibody or fragment thereof capable of specifically binding to a site on a mouse or human VE-cadherin, said site being within the about first 15 N-terminal amino acids of domain 1 of the VE-cadherin, wherein said antibody or fragment thereof is capable of inhibiting VE-cadherin mediated adherens junction formation in vitro but does not exert any significant or substantial effect on paracellular permeability in vitro, and 
 a pharmaceutically acceptable carrier or diluent. 
   
     
     
         32 . The method of  claim 31 , wherein said neoplastic disease comprises tumor growth or tumor metastasis. 
     
     
         33 . The method of  claim 31 , wherein the antibody or fragment thereof does not exert any significant or substantial effect on vascular permeability in vivo. 
     
     
         34 . The method of  claim 31 , wherein the antibody or fragment thereof inhibits formation of new adherens junctions without disturbing existing adherens junctions. 
     
     
         35 . The method of  claim 31 , wherein the antibody or fragment thereof is a single chain antibody, is humanized, is chimerized or is bispecific. 
     
     
         36 . The method of  claim 31 , wherein the antibody or fragment thereof is fused to a heterologous polypeptide. 
     
     
         37 . A method for reducing or inhibiting tumor vasculature in a mammal which comprises:
 administering a pharmaceutical composition to a mammal in need of reduction or inhibition of tumor vasculature in an amount effective to inhibit tumor blood vessel formation without adversely affecting existing vasculature, the pharmaceutical composition comprising:
 an isolated monoclonal antibody or fragment thereof capable of specifically binding to a site on a mouse or human VE-cadherin, said site being within the about first 15 N-terminal amino acids of domain 1 of the VE-cadherin, wherein said antibody or fragment thereof is capable of inhibiting VE-cadherin mediated adherens junction formation in vitro but does not exert any significant or substantial effect on paracellular permeability in vitro, and 
 a pharmaceutically acceptable carrier or diluent. 
   
     
     
         38 . The method of  claim 37 , wherein the antibody or fragment thereof does not exert any significant or substantial effect on vascular permeability in vivo. 
     
     
         39 . The method of  claim 37 , wherein the antibody or fragment thereof inhibits formation of new adherens junctions without disturbing existing adherens junctions. 
     
     
         40 . The method of  claim 37 , wherein the antibody or fragment thereof is a single chain antibody, is humanized, is chimerized or is bispecific. 
     
     
         41 . The method of  claim 37 , wherein tie antibody or fragment thereof is fused to a heterologous polypeptide.