Claims
- 1. A humanized anti-vascular endothelial growth factor (VEGF) antibody which binds human VEGF with a Kd value of no more than about 1×10−8M.
- 2. A humanized anti-vascular endothelial growth factor (VEGF) antibody which binds human VEGF with a Kd value of no more than about 5×10−9M.
- 3. A humanized anti-vascular endothelial growth factor (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-vascular endothelial growth factor (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 tumor growth by at least about 50% in tumor weight in vivo in nude mice transformed with human A673 rhabdomyosarcoma cells.
- 6. The humanized anti-VEGF antibody of claim 1 having a heavy chain variable domain comprising the following complementarity determining region (CDR) amino acid sequences: CDRH1 (GYX1FTX2YGMN, wherein X1 is T or D and X2 is N or H; SEQ ID NO: 130). CDRH2 (WINTYTGEPTYAADFKR; SEQ ID NO: 2) and CDRH3 (YPX1YYGX2SHWYFDV, wherein X1 is Y or H and X2 is S or T; SEQ ID NO: 131).
- 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: DIQX1,TQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKVLIYFTSSLHSGVPSRFSG SGSGTDFTLTISSLQPEDFATYYCQQYSTVPWTFGQGTKVEIKR (SEQ ID NO: 126), wherein X1 is M or L.
- 13. An anti-VEGF antibody heavy chain variable domain comprising the amino acid sequence: EVQLVESGGGLVQPGGSLRLSCAASGYX1FTX2YGMNWVRQAPGKGLEWVGWINTYTGEPTY AADFKRRFTFSLDTSKSTAYLQMNSLRAEDTAVYYCAKYPX3YYGX4SHWYFDVWGQGTLVTV SS (SEQ ID NO: 127), wherein X1 is T or D; X2 is N or H; X3 is Y or H and X4 is S or T.
- 14. A variant of a parent anti-vascular endothelial growth factor (VEGF) antibody, wherein said variant binds human VEGF and comprises an amino acid substitution in a complementarity determining region (CDR) 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 Kd value of no more than about 1×10−8M.
- 17. The variant of claim 14 which binds human VEGF with a Kd value of no more than about 5×10−9M.
- 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 Kd 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: 128)
- 24. The variant of claim 19 wherein the CDRH3 comprises the amino acid sequence: YPYYYGTSHWYFDV (SEQ ID NO: 129).
- 25. The variant of claim 14 wherein the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 118.
- 26. The variant of claim 25 further comprising the light chain variable domain amino acid sequence of SEQ ID NO: 126.
- 27. The variant of claim 26 comprising the light chain variable domain amino acid sequence of SEQ ID NO: 117.
- 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.
CROSS REFERENCES
[0001] This application is a continuation of co-pending U.S. application Ser. No. 09/056,160 filed Apr. 6, 1998, which claims benefit under 35 U.S.C. §119(e) of Provisional Application Serial No. 60/126,446, filed Apr. 7, 1997, and Serial No. 60/054,856 filed Aug. 6, 1997. The disclosure of both aforementioned Applications are expressly incorporated herein by reference.
Provisional Applications (2)
|
Number |
Date |
Country |
|
60126446 |
Apr 1997 |
US |
|
60054856 |
Aug 1997 |
US |
Continuations (1)
|
Number |
Date |
Country |
Parent |
09056160 |
Apr 1998 |
US |
Child |
10234671 |
Sep 2002 |
US |