Claims
- 1. An antibody having a first antigen binding site specific for a first VEGF receptor and a second antigen binding site specific for a second VEGF receptor.
- 2. The antibody of claim 1 wherein the first and second VEGF receptors are mammalian.
- 3. The antibody of claim 1 wherein the first and second VEGF receptors are human.
- 4. The antibody of claim 3 wherein the first and second VEGF receptors are selected from the group consisting of KDR, Flt-1 and Flt-4.
- 5. The antibody of claim 3 wherein the first VEGF receptor is KDR and the second VEGF receptor is Flt-1.
- 6. The antibody of claim 3 wherein the first VEGF receptor is KDR and the amino acid sequences of the complementarity determining regions (CDRs) of the first antigen binding site comprise:
SEQ ID NO: 1 at CDRH1; SEQ ID NO: 2 at CDRH2; SEQ ID NO: 3 at CDRH3; SEQ ID NO: 4 at CDRL1; SEQ ID NO: 5 at CDRL2; and SEQ ID NO: 6 at CDRL3.
- 7. The antibody of claim 3 wherein the first VEGF receptor is KDR and the amino acid sequences of the variable domains of the first antigen binding site comprise:
SEQ ID NO: 7 for the heavy-chain variable domain (VH); and SEQ ID NO: 8 for the light-chain variable domain (VL).
- 8. The antibody of claim 3 wherein the first VEGF receptor is KDR and the nucleotide sequences of the complementarity determining regions (CDRs) of the first antigen binding site comprise:
SEQ ID NO: 9 for CDRH1; SEQ ID NO: 10 for CDRH2; SEQ ID NO: 11 for CDRH3; SEQ ID NO: 12 for CDRL1; SEQ ID NO: 13 for CDRL2; and SEQ ID NO: 14 for CDRL3.
- 9. The antibody of claim 3 wherein the first VEGF receptor is KDR and the nucleotide sequences of the variable domains of the first antigen binding site comprise:
SEQ ID NO: 15 for the heavy-chain variable domain (VH); and SEQ ID NO: 16 for the light-chain variable domain (VL).
- 10. The antibody of claim 3 wherein the first VEGF receptor is KDR and the amino acid sequences of the complementarity determining regions (CDRs) of the first antigen binding site comprise:
SEQ ID NO: 1 for CDRH1; SEQ ID NO: 21 for CDRH2; SEQ ID NO: 3 for CDRH3; SEQ ID NO: 4 for CDRL1; SEQ ID NO: 5 for CDRL2; and SEQ ID NO: 6 for CDRL3.
- 11. The antibody of claim 3 wherein the first VEGF receptor is KDR and the amino acid sequences of the variable domains of the first antigen binding site comprise:
SEQ ID NO: 22 for the heavy-chain variable domain (VH); and SEQ ID NO: 23 for the light-chain variable domain (VL).
- 12. The antibody of claim 3 wherein the first VEGF receptor is KDR and the nucleotide sequences of the complementarity determining regions (CDRs) of the first antigen binding site comprise:
SEQ ID NO: 9 for CDRH1; SEQ ID NO: 24 for CDRH2; SEQ ID NO: 11 for CDRH3; SEQ ID NO: 12 for CDRL1; SEQ ID NO: 13 for CDRL2; and SEQ ID NO: 14 for CDRL3.
- 13. The antibody of claim 3 wherein the first VEGF receptor is KDR and the nucleotide sequences of the variable domains of the first antigen binding site comprise:
SEQ ID NO: 25 for the heavy-chain variable domain (VH); and SEQ ID NO: 26 for the light-chain variable domain (VL).
- 14. The antibody of claim 3 wherein the first VEGF receptor is KDR and the first antigen binding site comprises a set of amino acid sequences at CDRL1, CDRL2, CDRL3, CDRH1, CDRH2, and CDRH3, the set selected from the group consisting of the set of SEQ ID NOS:53, 54, 55, 65, 66, and 67, the set of SEQ ID NOS:56, 57, 58, 65, 66 and 67, the set of SEQ ID NOS:59, 60, 61, 65, 66, and 67, and the set of SEQ ID NOS:62, 63, 64, 68, 69 and 70.
- 15. The antibody of claim 3 wherein the first VEGF receptor is KDR and the first binding domain comprises a pair of VH and VL domains, the pairs selected from the group consisting of SEQ ID NOS:72 and 74, SEQ ID NOS:76 and 78, SEQ ID NOS:76 and 81, and SEQ ID NOS:83 and 85.
- 16. The antibody of claim 3 wherein the first VEGF receptor is KDR and the first antigen binding site comprises the set of amino acid sequences CDH1, CDRH2, and CDRH3 given by SEQ ID NOS: 65, 66, and 67, respectively, and a set of amino acid sequences at CDRL1, CDRL2, CDRL3 selected from the group consisting of the set of SEQ ID NOS:106, 107, and 108, the set of SEQ ID NOS:109, 110, and 111, the set of SEQ ID NOS:112, 113, and 114, the set of SEQ ID NOS:115, 116, and 117, the set of SEQ ID NOS:118, 119, and 120, the set of SEQ ID NOS:121, 122, and 123, the set of SEQ ID NOS:124, 125, and 126, the set of SEQ ID NOS:127, 128, and 129, the set of SEQ ID NOS:130, 131, and 132, and the set of SEQ ID NOS:133, 134, and 135.
- 17. The antibody of claim 3 wherein the first VEGF receptor is KDR, the VH domain of first binding domain comprises SEQ ID NO:76, and the VL domain of the first binding domain comprises a sequence selected from the group consisting of SEQ ID NO:87, SEQ ID NO:89, SEQ ID NO:91, SEQ ID NO:93, SEQ ID NO:95, SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO:101, SEQ ID NO:103, and SEQ ID NO:105.
- 18. The antibody of any one of claims 6 to 17 wherein the second VEGF receptor is Flt-1 and the second antigen binding site comprises the heavy chain and light chain variable domains of Mab 6.12 (ATCC No. PTA-3344).
- 19. The antibody of claim 3 wherein the first VEGF receptor is Flt-1 and the amino acid sequences of the complementarity determining regions (CDRs) of the first antigen binding site comprise:
SEQ ID NO: 35 at CDRH1; SEQ ID NO: 36 at CDRH2; SEQ ID NO: 37 at CDRH3; SEQ ID NO: 38 at CDRL1; SEQ ID NO: 39 at CDRL2; and SEQ ID NO: 40 at CDRL3.
- 20. The antibody of claim 3 wherein the first VEGF receptor is Flt-1 and the amino acid sequences of the variable domains of the first antigen binding site comprise:
SEQ ID NO: 41 for the heavy-chain variable domain (VH); and SEQ ID NO: 42 for the light-chain variable domain (VL).
- 21. The antibody of claim 3 wherein the first VEGF receptor is Flt-1 and the nucleotide sequences of the complementarity determining regions (CDRs) of the first antigen binding site comprise:
SEQ ID NO: 43 for CDRH1; SEQ ID NO: 44 for CDRH2; SEQ ID NO: 45 for CDRH3; SEQ ID NO: 46 for CDRL1; SEQ ID NO: 47 for CDRL2; and SEQ ID NO: 48 for CDRL3.
- 22. The antibody of claim 3 wherein the first VEGF receptor is Flt-1 and the nucleotide sequences of the variable domains of the first antigen binding site comprise:
SEQ ID NO: 49 for the heavy-chain variable domain (VH); and SEQ ID NO: 50 for the light-chain variable domain (VL).
- 23. The antibody of claim 3 wherein the first VEGF receptor is Flt-1 and the first antigen binding site comprises the heavy chain and light chain variable domains of Mab 6.12 (ATCC No. PTA-3344).
- 24. An antibody that binds specifically to an extracellular domain of a first VEGF receptor and an extracellular domain of a second VEGF receptor, wherein binding of the antibody to the first or the second VEGF receptor neutralizes activation of that VEGF receptor.
- 25. The antibody of claim 24 which blocks binding of VEGF.
- 26. The antibody of claim 24 which blocks receptor homodimerization.
- 27. The antibody of claim 24 which blocks receptor heterodimerization.
- 28. The antibody of claim 24 wherein the first and second VEGF receptors are selected from the group consisting of KDR, Flt-1 and Flt-4.
- 29. The antibody of claim 24 wherein the first VEGR receptor is KDR and the second VEGR receptor is Flt-1.
- 30. An antibody that binds specifically to an extracellular domain of a first VEGF receptor and an extracellular domain of a second VEGF receptor and reduces tumor growth.
- 31. The antibody of claim 29 wherein the first and second VEGF receptors are selected from the group consisting of KDR, Flt-1 and Flt-4.
- 32. The antibody of claim 29 wherein the first VEGR receptor is KDR and the second VEGR receptor is Flt-1.
- 33. A antibody that binds specifically to an extracellular domain of a first VEGF receptor and an extracellular domain of a second VEGF receptor and inhibits angiogenesis.
- 34. The antibody of claim 32 wherein the first and second VEGF receptors are selected from the group consisting of KDR, Flt-1 and Flt-4.
- 35. The antibody of claim 32 wherein the first VEGR receptor is KDR and the second VEGR receptor is Flt-1.
- 36. A method for making an antibody having a first antigen binding site comprising a first immunoglobulin heavy chain variable domain and a first immunoglobulin light chain variable domain that specifically binds to an extracellular domain of a first VEGF receptor, and a second antigen binding site comprising a second immunoglobulin heavy chain variable domain and a second immunoglobulin light chain variable domain that specifically binds to an extracellular domain of a second VEGF receptor, which comprises
a) coexpressing in a host cell
a recombinant DNA construct encoding a first polypeptide having the first immunoglobulin heavy chain variable domain located to the N terminus of the second immunoglobulin light chain variable domain, and a recombinant DNA construct encoding a second polypeptide having the second immunoglobulin heavy chain variable domain located to the N terminus of the first immunoglobulin light chain variable domain, for a time and in a manner sufficient to allow expression of the polypeptides and formation of the antibody; and b) recovering the antibody.
- 37. The method of claim 35 wherein the constructs are on the same DNA expression vector.
- 38. The method of claim 35 wherein the constructs are on different DNA expression vectors.
- 39. The method of claim 35 wherein the host cell is a bacterial cell, a yeast cell or a mammalian cell.
- 40. The method of claim 35 wherein the antibody is secreted from the host cell.
- 41. A method for neutralizing activation of a first VEGF receptor and a second VEGF receptor in a cell which comprises treating a cell with an antibody having a first antigen binding site specific for the first VEGF receptor and a second binding site specific for the second VEGF receptor in an amount sufficient to neutralize activation of the receptors.
- 42. A method for reducing tumor growth in a mammal in need thereof comprising treating the mammal with an antibody having a first antigen binding site specific for the first VEGF receptor and a second binding site specific for the second VEGF receptor in an amount effective to reduce tumor growth.
- 43. A method for inhibiting angiogenesis in a mammal in need thereof comprising treating the mammal with a bispecific antibody having a first antigen binding site specific for the first VEGF receptor and a second binding site specific for the second VEGF receptor in an amount effective to inhibit angiogenesis.
Parent Case Info
[0001] This application claims the benefit of U.S. Provisional Application Ser. No. 60/301,299, filed Jun. 26, 2001.
PCT Information
Filing Document |
Filing Date |
Country |
Kind |
PCT/US02/20332 |
6/26/2002 |
WO |
|
Provisional Applications (1)
|
Number |
Date |
Country |
|
60301299 |
Jun 2001 |
US |