Vegf/dll4 binding agents and uses thereof

Abstract

The present invention relates to VEGF-binding agents, DLL4-binding agents, VEGF/DLL4 bispecific binding agents, and methods of using the agents for treating diseases such as cancer. The present invention provides antibodies that specifically bind human VEGF, antibodies that specifically bind human DLL4, and bispecific antibodies that specifically bind human VEGF and/or human DLL4. The present invention further provides methods of using the agents to inhibit tumor growth. Also described are methods of treating cancer comprising administering a therapeutically effect amount of an agent or antibody of the present invention to a patient having a tumor or cancer.

1 - 89 . (canceled) 
     
     
         90 . An isolated polynucleotide encoding a modified immunoglobulin molecule, wherein the modified immunoglobulin molecule comprises:
 (a) a first antigen-binding site that specifically binds human VEGF; and   (b) a second antigen-binding site that specifically binds human DLL4;   wherein the first antigen-binding site comprises a heavy chain CDR1 comprising NYWMH (SEQ ID NO:17), a heavy chain CDR2 comprising DINPSNGRTSYKEKFKR (SEQ ID NO:18), and a heavy chain CDR3 comprising HYDDKYYPLMDY (SEQ ID NO:19);   wherein the second antigen-binding site comprises a heavy chain CDR1 comprising TAYYIH (SEQ ID NO:13) or AYYIH (SEQ ID NO:79), a heavy chain CDR2 comprising YIANYNRATNYNQKFKG (SEQ ID NO:14), YISSYNGATNYNQKFKG (SEQ ID NO:15), YIAGYKDATNYNQKFKG (SEQ ID NO:59), or YISNYNRATNYNQKFKG (SEQ ID NO:65), and a heavy chain CDR3 comprising RDYDYDVGMDY (SEQ ID NO:16); and   wherein both the first and second-antigen binding sites comprise a light chain having at least 95% sequence identity to SEQ ID NO:4 that lacks signal sequence MVLQTQVFISLLLWISGAYG (amino acid residues 1-20 of SEQ ID NO:4).   
     
     
         91 . The polynucleotide of  claim 90 , wherein both the first and second antigen-binding sites comprise a light chain comprising SEQ ID NO:4 that lacks signal sequence MVLQTQVFISLLLWISGAYG (amino acid residues 1-20 of SEQ ID NO:4). 
     
     
         92 . The polynucleotide of  claim 90 , wherein the light chain further comprises a light chain CDR1 comprising RASESVDNYGISFMK (SEQ ID NO:20), a light chain CDR2 comprising AASNQGS (SEQ ID NO:21), and light chain CDR3 comprising QQSKEVPWTFGG (SEQ ID NO:22). 
     
     
         93 . The polynucleotide of  claim 90 , wherein the heavy chain variable region of the second antigen-binding site comprises (i) a heavy chain CDR1 comprising TAYYIH (SEQ ID NO:13), (ii) a heavy chain CDR2 comprising YISNYNRATNYNQKFKG (SEQ ID NO:65), and (iii) a heavy chain CDR3 comprising RDYDYDVGMDY (SEQ ID NO:16). 
     
     
         94 . An isolated polynucleotide encoding a modified immunoglobulin molecule, wherein the modified immunoglobulin molecule comprises:
 (a) a first antigen-binding site that specifically binds human VEGF; and   (b) a second antigen-binding site that specifically binds human DLL4;   wherein the first antigen-binding site comprises a heavy chain CDR1 comprising NYWMH (SEQ ID NO:17), a heavy chain CDR2 comprising DINPSNGRTSYKEKFKR (SEQ ID NO:18), and a heavy chain CDR3 comprising HYDDKYYPLMDY (SEQ ID NO:19);   wherein the second antigen-binding site comprises a heavy chain CDR1 comprising TAYYIH (SEQ ID NO:13) or AYYIH (SEQ ID NO:79), a heavy chain CDR2 comprising YIX 1 X 2 YX 3 X 4 ATNYNQKFKG (SEQ ID NO:80), wherein X 1  is serine or alanine, X 2  is serine, asparagine, or glycine, X 3  is asparagine or lysine, and X 4  is glycine, arginine, or aspartic acid, and a heavy chain CDR3 comprising RDYDYDVGMDY (SEQ ID NO:16); and   wherein both the first and second-antigen binding sites comprise a light chain having at least 95% sequence identity to SEQ ID NO:4 that lacks signal sequence MVLQTQVFISLLLWISGAYG (amino acid residues 1-20 of SEQ ID NO:4).   
     
     
         95 . The polynucleotide of  claim 94 , wherein both the first and second antigen-binding sites comprise a light chain comprising SEQ ID NO:4 that lacks signal sequence MVLQTQVFISLLLWISGAYG (amino acid residues 1-20 of SEQ ID NO:4). 
     
     
         96 . The polynucleotide of  claim 94 , wherein the light chain further comprises a light chain CDR1 comprising RASESVDNYGISFMK (SEQ ID NO:20), a light chain CDR2 comprising AASNQGS (SEQ ID NO:21), and light chain CDR3 comprising QQSKEVPWTFGG (SEQ ID NO:22). 
     
     
         97 . The polynucleotide of  claim 94 , wherein the heavy chain variable region of the second antigen-binding site comprises (i) a heavy chain CDR1 comprising TAYYIH (SEQ ID NO:13), (ii) a heavy chain CDR2 comprising YISNYNRATNYNQKFKG (SEQ ID NO:65), and (iii) a heavy chain CDR3 comprising RDYDYDVGMDY (SEQ ID NO:16). 
     
     
         98 . A composition comprising the polynucleotide of  claim 90 . 
     
     
         99 . A composition comprising the polynucleotide of  claim 94 . 
     
     
         100 . A vector comprising the polynucleotide of  claim 90 . 
     
     
         101 . A vector comprising the polynucleotide of  claim 94 . 
     
     
         102 . An isolated cell comprising the vector of  claim 90 . 
     
     
         103 . An isolated cell comprising the vector of  claim 94 . 
     
     
         104 . An isolated polynucleotide encoding an antibody that specifically binds VEGF, wherein the antibody comprises:
 (a) a heavy chain having the sequence of SEQ ID NO:49 or SEQ ID NO:7; and   (b) a light chain having the sequence of SEQ ID NO:4 that lacks signal sequence MVLQTQVFISLLLWISGAYG (amino acid residues 1-20 of SEQ ID NO:4).   
     
     
         105 . The polynucleotide of  claim 104 , wherein the antibody comprises:
 (a) a heavy chain variable region having the sequence of SEQ ID NO:11; and   (b) a light chain variable region having the sequence of SEQ ID NO:12.   
     
     
         106 . A composition comprising the polynucleotide of  claim 104 . 
     
     
         107 . A composition comprising the polynucleotide of  claim 105 . 
     
     
         108 . A vector comprising the polynucleotide of  claim 104 . 
     
     
         109 . An isolated cell comprising the vector of  claim 104 .