Antibodies specific for urokinase-type plasminogen activator and methods of treating cancer

Patent metadata

Field	Value
Publication number	US-9255155-B2
Application number	US-201414162439-A
Country	US
Kind code	B2
Filing date	Jan 23, 2014
Priority date	Jan 31, 2013
Publication date	Feb 9, 2016
Grant date	Feb 9, 2016

Abstract

The present disclosure relates to antibodies specific for urokinase-type plasminogen activator (uPA). According to certain embodiments, the anti-uPA antibody specifically binds to the active form of uPA. In certain aspects, the anti-uPA antibody that specifically binds to active uPA binds specifically to the active form of human uPA (e.g., the antibody does not cross-react with active forms of uPA from non-human organisms). In certain aspects, an anti-uPA antibody of the present disclosure competes for specific binding to uPA with plasminogen activator inhibitor type 1 (PAI-1), where binding of the antibody to uPA results in internalization of a complex that includes the antibody, uPA, and urokinase-type plasminogen activator receptor (uPAR). Also provided are antibodies that specifically bind to uPA and compete for binding to uPA with a synthetic ligand of uPA. The disclosure also provides anti-uPA antibody conjugates and compositions (e.g., pharmaceutical formulations) comprising the antibodies and/or conjugates. Methods and kits related to the anti-uPA antibodies, conjugates, or formulations including the antibodies and/or conjugates are also provided.

First claim

What is claimed is:

1. A monoclonal antibody that competes for specific binding to urokinase-type plasminogen activator (uPA) with plasminogen activator inhibitor type 1 (PAI-1), wherein binding of the monoclonal antibody to uPA results in internalization of a complex comprising the antibody, uPA, and urokinase-type plasminogen activator receptor (uPAR).

2. The monoclonal antibody of claim 1 , wherein the monoclonal antibody competes for specific binding to uPA with an antibody comprising:
a heavy chain complementary determining region 1 (HCDR1) having the amino acid sequence of SEQ ID NO:1;
a heavy chain complementary determining region 2 (HCDR2) having the amino acid sequence of SEQ ID NO:2;
a heavy chain complementary determining region 3 (HCDR3) having the amino acid sequence of SEQ ID NO:3;
a light chain complementary determining region 1 (LCDR1) having the amino acid sequence of SEQ ID NO:4;
a light chain complementary determining region 2 (LCDR2) having the amino acid sequence of SEQ ID NO:5; and
a light chain complementary determining region 3 (LCDR3) having the amino acid sequence of SEQ ID NO:6.

3. The monoclonal antibody of claim 2 , wherein the monoclonal antibody comprises:
a heavy chain complementary determining region 1 (HCDR1) having the amino acid sequence of SEQ ID NO:1;
a heavy chain complementary determining region 2 (HCDR2) having the amino acid sequence of SEQ ID NO:2;
a heavy chain complementary determining region 3 (HCDR3) having the amino acid sequence of SEQ ID NO:3;
a light chain complementary determining region 1 (LCDR1) having the amino acid sequence of SEQ ID NO:4;
a light chain complementary determining region 2 (LCDR2) having the amino acid sequence of SEQ ID NO:5; and
a light chain complementary determining region 3 (LCDR3) having the amino acid sequence of SEQ ID NO:6.

4. The monoclonal antibody of claim 3 , wherein the monoclonal antibody comprises a heavy chain polypeptide comprising a variable region comprising the amino acid sequence set forth in SEQ ID NO:7.

5. The monoclonal antibody of claim 3 , wherein the monoclonal antibody comprises a light chain polypeptide comprising a variable region comprising the amino acid sequence set forth in SEQ ID NO:8.

6. The monoclonal antibody of claim 3 , wherein the monoclonal antibody comprises heavy and light chain polypeptides comprising variable regions comprising the amino acid sequences set forth in SEQ ID NO:7 and SEQ ID NO:8, respectively.

7. The monoclonal antibody of claim 1 , wherein the complex does not comprise PAI-1.

8. The monoclonal antibody of claim 1 , wherein the monoclonal antibody is selected from the group consisting of: an IgG, Fv, scFv, Fab, F(ab′)2, or Fab′.

9. A conjugate comprising:
a monoclonal antibody according to claim 1 ; and
an agent selected from the group consisting of: a therapeutic agent, and a labeling agent.

10. The conjugate of claim 9 , wherein the agent is a therapeutic agent.

11. The conjugate of claim 10 , wherein the therapeutic agent is a cytotoxic agent selected from the group consisting of: a radionuclide, a chemotherapeutic agent, and a toxin.

12. The conjugate of claim 9 , wherein the agent is a labeling agent.

13. The conjugate of claim 12 , wherein the labeling agent is an in vivo imaging agent.

14. A sterile pharmaceutical composition comprising:
a monoclonal antibody according to claim 1 ; and
a pharmaceutically acceptable carrier.

15. A method for treating cancer comprising the step of administering to a subject in need thereof a therapeutically effective amount of a monoclonal antibody according to claim 1 .

16. The method of claim 15 , wherein the cancer is selected from the group consisting of: prostate cancer, breast cancer, gastric cancer, colorectal cancer, esophageal cancer, renal cancer, endometrial cancer, and ovarian cancer.

17. The method of claim 16 , wherein the cancer is castration-resistant prostate cancer (CRPC).

18. A monoclonal antibody that specifically binds to urokinase-type plasminogen activator (uPA) and competes for binding to uPA with a synthetic ligand of uPA.

19. The isolated antibody of claim 18 , wherein the synthetic ligand is a synthetic peptide ligand.

20. The monoclonal antibody of claim 18 , wherein the ligand is p-aminobenzamidine.

21. The monoclonal antibody of claim 18 , wherein the antibody competes for specific binding to urokinase-type plasminogen activator (uPA) with an antibody comprising:
a heavy chain complementary determining region 1 (HCDR1) having the amino acid sequence of SEQ ID NO:1;
a heavy chain complementary determining region 2 (HCDR2) having the amino acid sequence of SEQ ID NO:2;
a heavy chain complementary determining region 3 (HCDR3) having the amino acid sequence of SEQ ID NO:3;
a light chain complementary determining region 1 (LCDR1) having the amino acid sequence of SEQ ID NO:4;
a light chain complementary determining region 2 (LCDR2) having the amino acid sequence of SEQ ID NO:5; and
a light chain complementary determining region 3 (LCDR3) having the amino acid sequence of SEQ ID NO:6.

22. The monoclonal antibody of claim 21 , wherein the monoclonal antibody comprises:
a heavy chain complementary determining region 1 (HCDR1) having the amino acid sequence of SEQ ID NO:1;
a heavy chain complementary determining region 2 (HCDR2) having the amino acid sequence of SEQ ID NO:2;
a heavy chain complementary determining region 3 (HCDR3) having the amino acid sequence of SEQ ID NO:3;
a light chain complementary determining region 1 (LCDR1) having the amino acid sequence of SEQ ID NO:4;
a light chain complementary determining region 2 (LCDR2) having the amino acid sequence of SEQ ID NO:5; and
a light chain complementary determining region 3 (LCDR3) having the amino acid sequence of SEQ ID NO:6.

23. The monoclonal antibody of claim 22 , wherein the monoclonal antibody comprises a heavy chain polypeptide comprising a variable region comprising the amino acid sequence set forth in SEQ ID NO:7.

24. The monoclonal antibody of claim 22 , wherein the monoclonal antibody comprises a light chain polypeptide comprising a variable region comprising the amino acid sequence set forth in SEQ ID NO:8.

25. The monoclonal antibody of claim 22 , wherein the monoclonal antibody comprises heavy and light chain polypeptides comprising variable regions comprising the amino acid sequences set forth in SEQ ID NO:7 and SEQ ID NO:8, respectively.

26. The monoclonal antibody of claim 18 , wherein binding of the antibody to uPA results in internalization of a complex comprising the antibody, uPA, and urokinase-type plasminogen activator receptor (uPAR).

27. The monoclonal antibody of claim 18 , wherein the complex does not comprise PAI-1.

28. The monoclonal antibody of claim 18 , wherein the antibody is selected from the group consisting of: an IgG, Fv, scFv, Fab, F(ab′)2, or Fab′.

29. A conjugate comprising:
a monoclonal antibody according to claim 18 ; and
an agent selected from: a therapeutic agent, and a labeling agent.

30. The conjugate of claim 29 , wherein the agent is a therapeutic agent.

31. The conjugate of claim 30 , wherein the therapeutic agent is a cytotoxic agent selected from the group consisting of: a radionuclide, a chemotherapeutic agent, and a toxin.

32. The c

Assignees

• Univ California
• Cytomx Therapeutics Inc

Inventors

• Sevillano Natalia
• Lebeau Aaron M
• Hostetter Daniel Robert
• Craik Charles S

Classifications

• A61K51/1075

  the antibody being against an enzyme · CPC title

• A61K49/0058

  Antibodies · CPC title

• A61K49/0032

  Methine dyes, e.g. cyanine dyes · CPC title

• C07K2317/565

  Complementarity determining region [CDR] · CPC title

• C07K16/40 Primary

  against enzymes · CPC title