Monoclonal antibodies specific to VEGF receptors and uses thereof

Abstract

Monoclonal antibodies that specifically bind to an extracellular domain of a flt-1 receptor and neutralize activation of the receptor are provided. In vitro and in vivo methods of using these antibodies are also provided.

What is claimed is:  
     
         1 . Nucleic acid molecules comprising a nucleic acid sequence that encodes an amino acid sequence wherein the amino acid sequence consists of the variable region of a monoclonal antibody that specifically binds to an extracellular domain of a flt-1 receptor and neutralizes activation of the receptor.  
     
     
         2 . The nucleic acid of  claim 1 , wherein the monoclonal antibody is produced by hybridoma cell line DC101 deposited as ATCC Accession No. HB 11534.  
     
     
         3 . The nucleic acid of  claim 1 , wherein the monoclonal antibody is produced by hybridoma cell line M25.18A1 deposited as ATCC Accession No. HB 12152.  
     
     
         4 . The nucleic acid of  claim 1 , wherein the monoclonal antibody is produced by hybridoma cell line M73.24 deposited as ATCC Accession No. HB 12153.  
     
     
         5 . Nucleic acid molecules comprising a nucleic acid sequence that encodes an amino acid sequence wherein the amino acid sequence consists of the hypervariable region of a monoclonal antibody that specifically binds to an extracellular domain of a flt-1 receptor and neutralizes activation of the receptor.  
     
     
         6  The nucleic acid of  claim 5 , wherein the monoclonal antibody is produced by hybridoma cell line DC101 deposited as ATCC Accession No. HB 11534.  
     
     
         7  The nucleic acid of  claim 5 , wherein the monoclonal antibody is produced by hybridoma cell line M25.18A1 deposited as ATCC Accession No. HB 12152.  
     
     
         8  The nucleic acid of  claim 5 , wherein the monoclonal antibody is produced by hybridoma cell line M73.24 deposited as ATCC Accession No. HB 12153.  
     
     
         9  A method for reducing tumor growth in a mammal in need thereof comprising treating the mammal with an effective amount of a monoclonal antibody which specifically binds to an extracellular domain of a flt-1 receptor and reduces tumor growth.  
     
     
         10 . The method of  claim 9 , wherein the antibody is produced by a hybridoma cell line.  
     
     
         11 . The method of  claim 10 , wherein the hybridoma cell line is deposited as ATCC Accession No. HB 11534.  
     
     
         12  A method for reducing tumor growth in a mammal in need thereof comprising treating the mammal with an effective amount of a chimeric antibody which comprises an amino acid sequence of a human antibody constant region and an amino acid sequence of a non-human antibody variable region, and which specifically binds to an extracellular domain of a flt-1 receptor and reduces tumor growth.  
     
     
         13  The method of  claim 12 , wherein the non-human variable region is murine.  
     
     
         14 . A method for reducing tumor growth in a mammal in need thereof comprising treating the mammal with an effective amount of a humanized antibody which comprises amino acid sequences of variable framework and constant regions from a human antibody, and an amino acid sequence of a non-human antibody hypervariable region, and which specifically binds to an extracellular domain of a flt-1 receptor and reduces tumor growth.  
     
     
         15 . The method of  claim 14 , wherein the amino acid sequence of the hypervariable region is murine.  
     
     
         16 . A method for inhibiting angiogenesis in a mammal in need thereof comprising treating the mammal with an effective amount of a monoclonal antibody which specifically binds to an extracellular domain of a flt-1 receptor and inhibits angiogenesis.  
     
     
         17  The method of  claim 16  wherein the antibody is produced by a hybridoma cell line  
     
     
         18  The method of  claim 17 , wherein the hybridoma cell line is deposited as ATCC Accession No. HB 11534.  
     
     
         19  A method for inhibiting angiogenesis in a mammal in need thereof comprising treating the mammal with an effective amount of a chimeric antibody which comprises an amino acid sequence of a human antibody constant region and an amino acid sequence of a non-human antibody variable region, and which specifically binds to an extracellular domain of a flt-1 receptor and inhibits angiogenesis.  
     
     
         20  A method of  claim 19 , wherein the non-human variable region is murine.  
     
     
         21 . A method for inhibiting angiogenesis in a mammal in need thereof comprising treating the mammal with an effective amount of a humanized antibody which comprises amino acid sequences of variable framework and constant regions from a human antibody, and an amino acid sequence of a non-human antibody hypervariable region, and which specifically binds to an extracellular domain of a flt-1 receptor and inhibits angiogenesis.  
     
     
         22  The method of  claim 21 , wherein the amino acid sequence of the hypervariable region is murine  
     
     
         23  The method of  claim 21 , wherein the hybridoma cell line is deposited as ATCC Accession No. HB 11534.  
     
     
         24  A method for reducing tumor growth in a mammal in need thereof comprising treating the mammal with an effective amount of a single chain antibody which specifically binds to an extracellular domain of a flt-1 receptor and reduces tumor growth.  
     
     
         25  A method for inhibiting angiogenesis in a mammal in need thereof comprising treating the mammal with an effective amount of a single chain antibody which specifically binds to an extracellular domain of a flt-1 receptor and inhibits angiogenesis  
     
     
         26 . A single chain antibody that specifically binds to an extracellular domain of a flt-1 receptor and neutralizes activation of the receptor.  
     
     
         27 . A single chain antibody that specifically binds to an extracellular domain of a flt-1 receptor and reduces tumor growth.  
     
     
         28 . A single chain antibody that specifically binds to an extracellular domain of a flt-1 receptor and inhibits angiogenesis.  
     
     
         29 . A process for preparing a polypeptide that comprises an amino acid sequence that specifically binds to an extracellular domain of a flt-1 receptor and neutralizes activation of the receptor, the process comprising: 
 culturing cells that express a nucleic acid molecule comprising a nucleic acid sequence that encodes an amino acid sequence wherein the amino acid sequence consists of the variable region of a monoclonal antibody that specifically binds to an extracellular domain of a flt-1 receptor and neutralizes activation of the receptor; and    isolating the polypeptide from the cultured cells.    
     
     
         30 . A process for preparing a polypeptide that comprises an amino acid sequence that specifically binds to an extracellular domain of a flt-1 receptor and neutralizes activation of the receptor, the process comprising: 
 culturing cells that express a nucleic acid molecule comprising a nucleic acid sequence that encodes an amino acid sequence wherein the amino acid sequence consists of the hypervariable region of a monoclonal antibody that specifically binds to an extracellular domain of a flt-1 receptor and neutralizes activation of the receptor; and    isolating the polypeptide from the cultured cells.    
     
     
         31 . A process for preparing chimerized monoclonal antibodies that specifically bind to an extracellular domain of a flt-1 receptor and neutralize activation of the receptor, the process comprising: 
 culturing cells that express a nucleic acid molecule comprising a nucleic acid sequence that encodes an amino acid sequence wherein the amino acid sequence consists of 
 (i) a variable region of a monoclonal antibody of a mammal other than a human wherein the variable region specifically binds to an extracellular domain of a flt-1 receptor and neutralizes activation of the receptor, and  
 (ii) a constant region of a human antibody; and isolating the chimerized monoclonal antibodies from the cultured cells.  
   
     
     
         32 . A process for preparing humanized monoclonal antibodies that specifically bind to an extracellular domain of a flt-1 receptor and neutralize activation of the receptor, the process comprising: 
 culturing cells that express a nucleic acid molecule comprising a nucleic acid sequence that encodes an amino acid sequence wherein the amino acid sequence consists of: 
 (i) a hypervariable region of a monoclonal antibody of a mammal other than a human wherein the hypervariable region specifically binds to an extracellular domain of a flt-1 receptor and neutralizes activation of the receptor,  
 (ii) a constant region of a human antibody, and  
 (iii) a variable region, other than the hypervariable region, substantially from a human antibody; and isolating the humanized monoclonal antibodies from the cultured cells.  
   
     
     
         33  A process for preparing a polypeptide that comprises an amino acid sequence that specifically binds to an extracellular domain of a flt-1 receptor and inhibits tumor growth in a mammal, the process comprising 
 culturing cells that express a nucleic acid molecule comprising a nucleic acid sequence that encodes an amino acid sequence wherein the amino acid sequence consists of the variable region of a monoclonal antibody that specifically binds to an extracellular domain of a flt-1 receptor and inhibits tumor growth in the mammal; and  
 isolating the polypeptide from the cultured cells.  
 
     
     
         34  A process for preparing a polypeptide that comprises an amino acid sequence that specifically binds to an extracellular domain of a flt-1 receptor and inhibits tumor growth in a mammal, the process comprising 
 culturing cells that express a nucleic acid molecule comprising a nucleic acid sequence that encodes an amino acid sequence wherein the amino acid sequence consists of the hypervariable region of a monoclonal antibody that specifically binds to an extracellular domain of a flt-1 receptor and inhibits tumor growth in the mammal; and  
 isolating the polypeptide from the cultured cells.  
 
     
     
         35 . A process for preparing chimerized monoclonal antibodies that specifically bind to an extracellular domain of a flt-1 receptor and inhibit tumor growth in a recipient mammal, the process comprising: 
 culturing cells that express a nucleic acid molecule comprising a nucleic acid sequence that encodes an amino acid sequence wherein the amino acid sequence consists of 
 (i) a variable region of a monoclonal antibody of a mammal other than a human wherein the variable region specifically binds to an extracellular domain of a flt-1 receptor and inhibits tumor growth in the recipient mammal, and  
 (ii) a constant region of a human antibody; and isolating the chimerized monoclonal antibodies from the cultured cells.  
   
     
     
         36 . A process for preparing humanized monoclonal antibodies that specifically bind to an extracellular domain of a flt-1 receptor and inhibit tumor growth in a recipient mammal, the process comprising 
 culturing cells that express a nucleic acid molecule comprising a nucleic acid sequence that encodes an amino acid sequence wherein the amino acid sequence consists of 
 (i) a hypervariable region of a monoclonal antibody of a mammal other than a human wherein the hypervariable region specifically binds to an extracellular domain of a flt-1 receptor and inhibits tumor growth in the recipient mammal,  
 (ii) a constant region of a human antibody, and  
 (iii) a variable region, other than the hypervariable region, substantially from a human antibody; and isolating the humanized monoclonal antibodies from the cultured cells.  
   
     
     
         37  A chimerized monoclonal antibody that specifically binds to an extracellular domain of a flt-1 receptor and neutralizes activation of the receptor.  
     
     
         38  A cell line producing the antibody of  claim 37   
     
     
         39  A composition comprising the antibody of  claim 37  and a pharmaceutically acceptable carrier  
     
     
         40 . The composition of  claim 39  further comprising a chemotherapeutic agent and a pharmaceutically acceptable carrier  
     
     
         41  A chimerized monoclonal antibody that specifically binds to an extracellular domain of a flt-1 receptor and reduces tumor growth.  
     
     
         42 . A cell line producing the antibody of  claim 41 .  
     
     
         43 . A composition comprising the antibody of  claim 41  and a pharmaceutically acceptable carrier.  
     
     
         44 . The composition of  claim 43  further comprising a chemotherapeutic agent and a pharmaceutically acceptable carrier.  
     
     
         45  A humanized monoclonal antibody that specifically binds to an extracellular domain of a flt-1 receptor and neutralizes activation of the receptor  
     
     
         46  A cell line producing the antibody of  claim 45   
     
     
         47 . A composition comprising the antibody of  claim 45  and a pharmaceutically acceptable carrier  
     
     
         48 . The composition of  claim 47  further comprising a chemotherapeutic agent and a pharmaceutically acceptable carrier.  
     
     
         49 . A humanized monoclonal antibody that specifically binds to an extracellular domain of a flt-1 receptor and reduces tumor growth  
     
     
         50 . A cell line producing the antibody of  claim 49 .  
     
     
         51 . A composition comprising the antibody of  claim 49  and a pharmaceutically acceptable carrier.  
     
     
         52 . The composition of claim  51  further comprising a chemotherapeutic agent and a pharmaceutically acceptable carrier.