Anti-vegf antibodies

Abstract

Humanized and variant anti-VEGF antibodies and various uses therefor are disclosed. The anti-VEGF antibodies have strong binding affinities for VEGF; inhibit VEGF-induced proliferation of endothelial cells in vitro; and inhibit tumor growth in vivo.

What is claimed is:  
     
         1 . A humanized anti-VEGF antibody which binds human VEGF with a K d  value of no more than about 1×10 −8 M.  
     
     
         2 . A humanized anti-VEGF antibody which binds human VEGF with a K d  value of no more than about 5×10 −9 M.  
     
     
         3 . A humanized anti-VEGF antibody which has an ED50 value of no more than about 5 nM for inhibiting VEGF-induced proliferation of endothelial cells in vitro.  
     
     
         4 . A humanized anti-VEGF antibody which inhibits VEGF-induced angiogenesis in vivo.  
     
     
         5 . The humanized anti-VEGF antibody of  claim 4  wherein 5 mg/kg of the antibody inhibits at least about 50% of tumor growth in an A673 in vivo tumor model.  
     
     
         6 . The humanized anti-VEGF antibody of  claim 1  having a heavy chain variable domain comprising the following hypervariable region amino acid sequences: CDRH1 (GYX 1 FTX 2 YGMN, wherein X 1  is T or D and X 2  is N or H; SEQ ID NO:128), CDRH2 (WINTYTGEPTYAADFKR; SEQ ID NO:2) and CDRH3 (YPX 1 YYGX 2 SHWYFDV, wherein X 1  is Y or H and X 2  is S or T; SEQ ID NO:129).  
     
     
         7 . The humanized anti-VEGF antibody of  claim 6  comprising the amino acid sequence of SEQ ID NO:7.  
     
     
         8 . The humanized anti-VEGF antibody of  claim 6  having a heavy chain variable domain comprising the following hypervariable region amino acid sequences: CDRH1 (GYTFTNYGMN; SEQ ID NO:1), CDRH2 (WINTYTGEPTYAADFKR; SEQ ID NO:2) and CDRH3 (YPHYYGSSHWYFDV; SEQ ID NO:3).  
     
     
         9 . The humanized anti-VEGF antibody of  claim 1  having a light chain variable domain comprising the following hypervariable region amino acid sequences: CDRL1 (SASQDISNYLN; SEQ ID NO:4), CDRL2 (FTSSLHS; SEQ ID NO:5) and CDRL3 (QQYSTVPWT; SEQ ID NO:6).  
     
     
         10 . The humanized anti-VEGF antibody of  claim 9  comprising the amino acid sequence of SEQ ID NO:8.  
     
     
         11 . The humanized anti-VEGF antibody of  claim 1  having a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:7 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO:8.  
     
     
         12 . An anti-VEGF antibody light chain variable domain comprising the amino acid sequence: DIQX 1 TQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKVLIYFTSSLHSGVPSRFS GSGSGTDFTLTISSLQPEDFATYYCQQYSTVPWTFGQGTKVEIKR (SEQ ID NO:124), wherein X 1  is M or L.  
     
     
         13 . An anti-VEGF antibody heavy chain variable domain comprising the amino acid sequence: EVQLVESGGGLVQPGGSLRLSCAASGYX 1 FTX 2 YGMNWVRQAPGKGLEWVGWINTYTGEPT YAADFKRRFTFSLDTSKSTAYLQMNSLRAEDTAVYYCAKYPX 3 YYGX 4 SHWYFDVWGQGTLV TVSS (SEQ ID NO:125), wherein X 1  is T or D; X 2  is N or H; X 3  is Y or H and X 4  is S or T.  
     
     
         14 . A variant of a parent anti-VEGF antibody, wherein said variant binds human VEGF and comprises an amino acid substitution in a hypervariable region of a heavy chain variable domain of said parent antibody.  
     
     
         15 . The variant of  claim 14  wherein said parent antibody is a human or humanized antibody.  
     
     
         16 . The variant of  claim 14  which binds human VEGF with a K d  value of no more than about 1×10 −8 M.  
     
     
         17 . The variant of  claim 14  which binds human VEGF with a K d  value of no more than about 5×10 −9 M.  
     
     
         18 . The variant of  claim 14  wherein the substitution is in CDRH1 of the heavy chain variable domain.  
     
     
         19 . The variant of  claim 14  wherein the substitution is in CDRH3 of the heavy chain variable domain.  
     
     
         20 . The variant of  claim 14  which has amino acid substitutions in both CDRH1 and CDRH3.  
     
     
         21 . The variant of  claim 14  which binds human VEGF with a K d  value less than that of said parent antibody.  
     
     
         22 . The variant of  claim 14  which has an ED50 value for inhibiting VEGF-induced proliferation of endothelial cells in vitro which is at least about 10 fold lower than that of said parent antibody.  
     
     
         23 . The variant of  claim 18  wherein the CDRH1 comprises the amino acid sequence: GYDFTHYGMN (SEQ ID NO:126)  
     
     
         24 . The variant of  claim 19  wherein the CDRH3 comprises the amino acid sequence: YPYYYGTSHWYFDV (SEQ ID NO:127).  
     
     
         25 . The variant of  claim 14  wherein the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO:116.  
     
     
         26 . The variant of  claim 25  further comprising the light chain variable domain amino acid sequence of SEQ ID NO:124.  
     
     
         27 . The variant of  claim 26  comprising the light chain variable domain amino acid sequence of SEQ ID NO:115.  
     
     
         28 . The humanized anti-VEGF antibody of  claim 1  which is a full length antibody.  
     
     
         29 . The humanized anti-VEGF antibody of  claim 28  which is a human IgG.  
     
     
         30 . The humanized anti-VEGF antibody of  claim 1  which is an antibody fragment.  
     
     
         31 . The antibody fragment of  claim 30  which is a Fab.  
     
     
         32 . A composition comprising the humanized anti-VEGF antibody of  claim 1  and a pharmaceutically acceptable carrier.  
     
     
         33 . A composition comprising the variant anti-VEGF antibody of  claim 14  and a pharmaceutically acceptable carrier.  
     
     
         34 . Isolated nucleic acid encoding the antibody of  claim 1 .  
     
     
         35 . A vector comprising the nucleic acid of  claim 34 .  
     
     
         36 . A host cell comprising the vector of  claim 35 .  
     
     
         37 . A process of producing a humanized anti-VEGF antibody comprising culturing the host cell of  claim 36  so that the nucleic acid is expressed.  
     
     
         38 . The process of  claim 37  further comprising recovering the humanized anti-VEGF antibody from the host cell culture.  
     
     
         39 . A method for inhibiting VEGF-induced angiogenesis in a mammal comprising administering a therapeutically effective amount of the humanized anti-VEGF antibody of  claim 1  to the mammal.  
     
     
         40 . The method of  claim 39  wherein the mammal is a human.  
     
     
         41 . The method of  claim 39  wherein the mammal has a tumor.  
     
     
         42 . The method of  claim 39  wherein the mammal has a retinal disorder.