Inhibition of tumor metastasis by anti-neuropilin 2 antibodies

1. A method for inhibiting lymphatic endothelial cell migration, comprising administering to a mammalian subject in need an effective amount of an anti-neuropilin2B (anti-Nrp2B) antibody, or an antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment:

i) comprises complementarity determining region light chain (CDRL)1, CDRL2, and CDRL3 sequences and complementarity determining region heavy chain (CDRH)1, CDRH2, CDRH3 sequences of an antibody selected from the group consisting of YW68.4.2 and YW68.4.2.36;

ii) is a variant of the antibody of (i) having one to five alterations in the framework regions, wherein the light chain framework regions comprise residues 1-23, 35-49, 57-88 and 98-107 or 1-25, 33-49, 53-90 and 97-107 and the heavy chain framework regions comprise residues 1-30, 36-49, 66-94 and 103-113 or 1-25, 33-52, 56-95 and 102-113; or

iii) is an affinity matured variant of the antibody of (i).

2. The method of claim 1 wherein said mammalian subject is a human patient.

3. The method of claim 2 wherein said human patient has been diagnosed with cancer.

4. The method of claim 3 wherein said human patient has developed or is at risk of developing tumor metastasis.

5. The method of claim 4 wherein said metastasis is in the lymphatic system.

6. The method of claim 4 wherein said metastasis is in a distant organ.

7. The method of claim 3 wherein said cancer is selected from the group consisting of carcinoma, lymphoma, blastoma, sarcoma, and leukemia.

8. The method of claim 3 wherein said cancer is selected from the group consisting of squamous cell cancer, small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, squamous carcinoma of the lung, cancer of the peritoneum, hepatocellular cancer, gastric cancer, gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, colorectal cancer, endometrial or uterine carcinoma, salivary gland carcinoma, kidney or renal cancer, liver cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma and various types of head and neck cancer, B-cell lymphoma, chronic lymphocytic leukemia (CLL); acute lymphoblastic leukemia (ALL); Hairy cell leukemia; chronic myeloblastic leukemia; post-transplant lymphoproliferative disorder (PTLD), abnormal vascular proliferation associated with phakomatoses, edema associated with brain tumors, and Meigs' syndrome.

9. The method of claim 8 wherein said B-cell lymphoma is selected from the group consisting of low grade/follicular non-Hodgkin's lymphoma (NHL); small lymphocytic (SL) NHL; intermediate grade/follicular NHL; intermediate grade diffuse NHL; high grade immunoblastic NHL; high grade lymphoblastic NHL; high grade small non-cleaved cell NHL; bulky disease NHL; mantle cell lymphoma; AIDS-related lymphoma; and Waldenstrom's Macroglobulinemia.

10. The method of claim 1 , wherein said variant is an affinity matured variant.

11. The method of claim 1 , wherein the anti-Nrp2B antibody, or an antigen-binding fragment thereof:

i) comprises the heavy and light chain variable region sequence of an antibody selected from the group consisting of YW68.4.2 and YW68.4.2.36;

ii) is a variant of the antibody of (i) having one to five alterations in the framework regions, wherein the light chain framework regions comprise residues 1-23, 35-49, 57-88 and 98-107 or 1-25, 33-49, 53-90 and 97-107 and the heavy chain framework regions comprise residues 1-30, 36-49, 66-94 and 103-113 or 1-25, 33-52, 56-95 and 102-113; or

iii) is an affinity matured variant of the antibody of (i).

12. The method of claim 11 , wherein the anti-Nrp2B antibody or antigen-binding fragment comprises the sequence of YW68.4.2 (SEQ ID NO: 1) or the sequence of YW68.4.2.36 (SEQ ID NO: 2).

13. The method of claim 12 , wherein the anti-Nrp2B antibody comprises the sequence of YW68.4.2.36 (SEQ ID NO: 1), or an antigen-binding fragment thereof.

14. The method of claim 11 , wherein the anti-Nrp2B antibody is selected from the group consisting of YW68.4.2 and YW68.4.2.36 and antigen-binding fragments thereof.

15. The method of claim 14 , wherein the anti-Nrp2B antibody is YW68.4.2.36 (SEQ ID NO: 1), or an antigen-binding fragment thereof.

16. The method of claim 1 , wherein said antibody, antigen-binding fragment or variant, is chimeric or humanized.

17. The method of claim 1 , wherein said antigen-binding fragment is selected from the group consisting of Fab, F(ab′) 2 , and scFv fragments.

18. The method of claim 1 , wherein the anti-Nrp2B antibody, or antigen-binding fragment or variant is bispecific.

19. The method of claim 18 , wherein the anti-Nrp2B antibody, or antigen-binding fragment or variant additionally binds to VEGF.

20. A method for inhibiting tumoral lymphangiogenesis, comprising administering to a tumor-bearing mammalian subject an effective amount of an anti-neuropilin2B (anti-Nrp2B) antibody, or an antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment:

i) comprises complementarity determining region light chain (CDRL)1, CDRL2, and CDRL3 sequences and complementarity determining region heavy chain (CDRH)1, CDRH2, CDRH3 sequences of an antibody selected from the group consisting of YW68.4.2 and YW68.4.2.36;

ii) is a variant of the antibody of (i) having one to five alterations in the framework regions, wherein the light chain framework regions comprise residues 1-23, 35-49, 57-88 and 98-107 or 1-25, 33-49, 53-90 and 97-107 and the heavy chain framework regions comprise residues 1-30, 36-49, 66-94 and 103-113 or 1-25, 33-52, 56-95 and 102-113; or

iii) is an affinity matured variant of the antibody of (i).

21. A method for treating tumor metastasis, comprising administering to a tumor-bearing mammalian subject an effective amount of an anti-neuropilin2B (anti-Nrp2B) antibody, or an antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment:

i) comprises complementarity determining region light chain (CDRL)1, CDRL2, and CDRL3 sequences and complementarity determining region heavy chain (CDRH)1, CDRH2, CDRH3 sequences of an antibody selected from the group consisting of YW68.4.2 and YW68.4.2.36;

ii) is a variant of the antibody of (i) having one to five alterations in the framework regions, wherein the light chain framework regions comprise residues 1-23, 35-49, 57-88 and 98-107 or 1-25, 33-49, 53-90 and 97-107 and the heavy chain framework regions comprise residues 1-30, 36-49, 66-94 and 103-113 or 1-25, 33-52, 56-95 and 102-113; or

iii) is an affinity matured variant of the antibody of (i).

22. The method of claim 20 or claim 21 wherein said mammalian subject is a human cancer patient.

23. The method of claim 22 wherein said human cancer patient has developed or is at risk of developing tumor metastasis.

24. The method of claim 23 wherein said tumor metastasis is in the lymphatic system.

25. The method of claim 23 wherein said tumor metastasis is in a distant organ.

26. The method of claim 22 wherein said cancer is selected from the group consisting of carcinoma, lymphoma, blastoma, sarcoma, and leukemia.

27. The method of claim 22 wherein said cancer is selected from the group consisting of squamous cell cancer, small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, squamous carcinoma of the lung, cancer of the peritoneum, hepatocellular cancer, gastric cancer, gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, colorectal cancer, endometrial or uterine carcinoma, salivary gland carcinoma, kidney or renal cancer, liver cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma and various types of head and neck cancer, B-cell lymphoma, chronic lymphocytic leukemia (CLL); acute lymphoblastic leukemia (ALL); Hairy cell leukemia; chronic myeloblastic leukemia; post-transplant lymphoproliferative disorder (PTLD), abnormal vascular proliferation associated with phakomatoses, edema associated with brain tumors, and Meigs' syndrome.

28. The method of claim 27 wherein said B-cell lymphoma is selected from the group consisting of low grade/follicular non-Hodgkin's lymphoma (NHL); small lymphocytic (SL) NHL; intermediate grade/follicular NHL; intermediate grade diffuse NHL; high grade immunoblastic NHL; high grade lymphoblastic NHL; high grade small non-cleaved cell NHL; bulky disease NHL; mantle cell lymphoma; AIDS-related lymphoma; and Waldenstrom's Macroglobulinemia.

29. The method of claim 23 , wherein said variant is an affinity matured variant.

30. The method of claim 23 , wherein the anti-Nrp2B antibody, or an antigen-binding fragment thereof:

i) comprises the heavy and light chain variable region sequence of an antibody selected from the group consisting of YW68.4.2 and YW68.4.2.36;

ii) is a variant of the antibody of (i) having one to five alterations in the framework regions, wherein the light chain framework regions comprise residues 1-23, 35-49, 57-88 and 98-107 or 1-25, 33-49, 53-90 and 97-107 and the heavy chain framework regions comprise residues 1-30, 36-49, 66-94 and 103-113 or 1-25, 33-52, 56-95 and 102-113; or

iii) is an affinity matured variant of the antibody of (i).

31. The method of claim 30 , wherein the anti-Nrp2B antibody or antigen-binding fragment comprises the sequence of YW68.4.2 (SEQ ID NO: 1) or the sequence of YW68.4.2.36 (SEQ ID NO: 2).

32. The method of claim 31 , wherein the anti-Nrp2B antibody comprises the sequence of YW68.4.2.36 (SEQ ID NO: 1), or an antigen-binding fragment thereof.

33. The method of claim 30 , wherein the anti-Nrp2B antibody is selected from the group consisting of YW68.4.2 and YW68.4.2.36 and antigen-binding fragments thereof.

34. The method of claim 33 , wherein the anti-Nrp2B antibody is YW68.4.2.36 (SEQ ID NO: 1), or an antigen-binding fragment thereof.

35. The method of claim 23 , wherein said antibody, antigen-binding fragment or variant, is chimeric or humanized.

36. The method of claim 23 , wherein said antigen-binding fragment is selected from the group consisting of Fab, F(ab′) 2 , and scFv fragments.

37. The method of claim 23 , wherein the anti-Nrp2B antibody, or antigen-binding fragment or variant is bispecific.

38. The method of claim 37 , wherein the anti-Nrp2B antibody, or antigen-binding fragment or variant additionally binds to VEGF.