CARBONIC ANHYDRASE IX (G250) ANITBODIES AND METHODS OF USE THEREOF

• US 20090068095A1
• Filed: 12/04/2006
• Published: 03/12/2009
• Est. Priority Date: 12/02/2005
• Status: Active Grant

• Current Assignee: Dana-Farber Cancer Institute
• Original Assignee: Dana-Farber Cancer Institute
• Inventors: Lo, Agnes, Marasco, Wayne A., Xu, Chen
• Granted Patent: US 8,466,263 B2
• Time in Patent Office: Days
• US Class Current: 424/1.490
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 38/2013 IL-2
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 51/1045 against animal or human tumor cells or tumor cell determinants
• CPC Class Codes: A61K 51/1075 the antibody being against an enzyme
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/73 Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding

Abstract

The invention provides scFv antibodies and monoclonal antibodies that bind to and decrease an activity of Carbonic Anhydrase IX (G250). Also provided are methods of treating and/or preventing cancer, such as renal clear cell cancer. Also provided are methods of identifying a carbonic anhydrase IX (G250) protein. The invention additionally provides methods of modifying immune effector cells, and the immune effector cells modified thereby.

Claims

1. An isolated human monoclonal antibody that immunospecifically binds to a carbonic anhydrase IX (G250) protein.

2. The antibody of claim 1, wherein said monoclonal antibody binds to the carbonic anhydrase (CA) domain of carbonic anhydrase IX (G250).

3. The antibody of claim 1, wherein said monoclonal antibody reduces carbonic anhydrase activity of said protein.

4. The antibody of claim 1, wherein said antibody has:

(a) a heavy chain with a CDR comprising amino acids 99 to 111 of SEQ ID NO;

1; and

(b) a light chain with a CDR comprising amino acids 91 to 102 of SEQ ID NO;

2.

5. The antibody of claim 1, wherein said antibody has a heavy chain with a CDR1 comprising an amino sequence selected from the group consisting of SYAMS, XYAMX, and SYXMX.

6. The antibody of claim 1, wherein said antibody has a heavy chain with a CDR2 comprising an amino sequence AISXXGGXTXXADSVKG or AISGSGGSTTTADSVKG.

7. The antibody of claim 1, wherein said antibody has a heavy chain with a CDR3 comprising an amino sequence NGNYRGSLXAFDI.

8. The antibody of claim 1, wherein said antibody has a light chain with a CDR1 comprising an amino sequence TGSSSNIGAGYDVH.

9. The antibody of claim 1, wherein said antibody has a light chain with a CDR2 comprising an amino sequence GNNNRPS.

10. The antibody of claim 1, wherein said antibody has a light chain with a CDR3 comprising an amino sequence QSYDSSLSAWVV.

17. A nucleic acid sequence comprising:

(a) a first nucleic acid encoding the antibody of claim 1; and

(b) a second nucleic acid encoding an IL-2 polypeptide.

18. A nucleic acid sequence comprising:

(a) a first nucleic acid encoding the antibody of claim 1; and

(b) a second nucleic acid encoding a zeta chain of the T-cell receptor complex.

19. The nucleic acid sequence of claim 18, further comprising a third nucleic acid sequence encoding a CD28 polypeptide.

20. A method of preventing or treating cancer, the method comprising administering to a person at risk or suffering from cancer, a therapeutically effective amount of a first antibody comprising the monoclonal antibody of claim 1.

21. The method of claim 20, wherein said cancer is renal cancer.

22. The method of claim 21, wherein said renal cancer is renal clear cell cancer.

23. The method of claim 20, further comprising administering a second antibody, wherein said first and second antibodies do not bind to the same epitope.

24. The method of claim 23, wherein said second antibody immunospecifically binds to a carbonic anhydrase IX (G250) protein.

25. The method of claim 23, wherein said second antibody is selected from the group consisting of Avastin, Erbitux, Humira, Xolair, Zavalin, Campath, Mylotarg, Herceptin, Remicaide, Simulect, Synagis, Zenapax, Rituxan, Panorex, ReoPro, Oncoscint, and OKT3.

26. The method of claim 20, further comprising administering a small molecule having an antineoplastic activity.

27. The method of claim 20, further comprising administering a cytokine.

28. The method of claim 20, wherein said cytokine is selected from the group consisting of IL-2, GM-CSF, IL-12, and TNF-alpha.

29. The monoclonal antibody of claim 1, or a functional fragment thereof wherein said antibody contains a H310A and a H435Q mutation when numbered in accordance with the Wu and Kabat numbering system.

30. A fusion protein comprising the monoclonal antibody of claim 1, or a functional fragment thereof, operably linked to an IL-2 polypeptide.

31. A fusion protein comprising the monoclonal antibody of claim 1, or a functional fragment thereof, operably linked to an IgG molecule.

32. A fusion protein comprising the monoclonal antibody of claim 1, or a functional fragment thereof, operably linked to a zeta chain of the T-cell receptor complex

33. The fusion protein of claim 32, further comprising a CD28 polypeptide.

34. A minibody comprising the binding domain of the monoclonal antibody of claim 1, or a functional fragment thereof, and a constant region of an immunoglobulin or fragment thereof.

35. A composition comprising the monoclonal antibody of claim 1 and a carrier.

36. The composition of claim 35, further comprising IL-2.

37. A kit comprising, in one or more containers, the composition of claim 35.

38. A method of quantitating the expression of a protein on or in a mammalian cell, comprising the steps of:

(a) providing a mammalian cell suspected of expressing a carbonic anhydrase IX (G250) protein;

(b) contacting said cell with the antibody of claim 1 under conditions where the carbonic anhydrase IX (G250) protein and the antibody are capable of forming a complex; and

(c) detecting the amount of complex formation,

thereby quantitating the expression of said carbonic anhydrase IX (G250) protein.

39. A method of modifying an immune effector cell, comprising:

(a) providing an immune effector cell obtained from a mammalian subject having renal cancer;

(b) contacting said immune effector cell with a nucleic acid encoding the antibody of claim 1 or a fragment thereof,

thereby modifying said immune effector cell.

40. The method of claim 39, wherein said immune effector cell is a T cell.

41. The method of claim 39, wherein said nucleic acid comprises a vector.

42. The method of claim 41, wherein said vector is a retroviral vector.

43. An isolated modified immune effector cell educated by the method of claim 39.

44. A method of preventing or treating cancer, comprising administering to a person at risk or suffering from cancer the modified immune effector cell of claim 43.

45. The method of claim 44, wherein said cancer is renal cancer.

46. The method of claim 45, wherein said renal cancer is renal clear cell cancer.

47. The method of claim 44, further comprising administering an anti-neoplastic agent.

48. The method of claim 44, further comprising administering a cytokine selected from the group consisting of IL-2, GM-CSF, IL-12, and TNF-alpha.

49. A method of detecting a carbonic anhydrase IX (G250) protein, comprising the steps of:

(a) providing a first detection means comprising the antibody of claim 1 operably linked to a support means;

(b) contacting said first detection means with a biological sample suspected of containing a carbonic anhydrase IX (G250) protein under conditions where the carbonic anhydrase IX (G250) protein and the first detection means are capable of forming a complex; and

(c) detecting the amount of complex formation,

thereby detecting said carbonic anhydrase IX (G250) protein.

52. A non-invasive method of detecting a tumor in a subject, comprising the steps of:

(a) administering to the subject the antibody of claim 1, linked to a detectable moiety;

(b) allowing for the localization of said antibody in the tumor; and

(c) detecting the detectable moiety, thereby detecting the tumor in the subject.

53. The method of claim 52, wherein said detectable moiety comprises a radioactive element.

54. The method of claim 52, wherein said detecting is performed by positron emission tomography.

11. An antibody complex comprising:

a) a first fully human monoclonal antibody that binds to a carbonic anhydrase IX (G250) protein, operably linked to

b) a second fully human monoclonal antibody that binds to a carbonic anhydrase IX (G250) protein, wherein said antibody complex reduces carbonic anhydrase activity.

12. The antibody of claim 11, wherein said first and second antibodies do not bind to the same epitope.

13. An isolated scFv antibody, wherein said scFv antibody binds to a carbonic anhydrase IX (G250) protein.

14. An isolated scFv antibody, wherein said scFv antibody binds to the carbonic anhydrase (CA) domain of a carbonic anhydrase IX (G250) protein.

15. The scFv antibody of claim 14, wherein said binding affinity is from about 10⁻

6 M to about 10<sup>−

12</sup> M.

16. The scFv antibody of claim, 14 wherein said scFv antibody has the sequence of SEQ ID NO:

3-50.

55. The isolated antibody of claim 14, wherein said antibody reduces carbonic anhydrase activity of said protein.

50. A method of detecting a carbonic anhydrase IX (G250) protein, comprising the steps of:

(a) contacting a biological sample suspected of containing a carbonic anhydrase IX (G250) protein with a first detection means comprising a first fully human monoclonal antibody that binds to the carbonic anhydrase (CA) domain of a carbonic anhydrase IX (G250) protein under conditions where the carbonic anhydrase IX (G250) protein and the first detection means are capable of forming a complex;

(b) contacting said biological sample with a second detection means comprising a second fully human monoclonal antibody that binds to the carbonic anhydrase (CA) domain of a carbonic anhydrase IX (G250) protein under conditions where the carbonic anhydrase IX (G250) protein and the second detection means are capable of forming a complex, wherein said first and second antibodies do not bind to the same epitope; and

(c) detecting the amount of complex formed between the carbonic anhydrase IX (G250) protein and the second detection means,

thereby detecting said carbonic anhydrase IX (G250) protein.

51. The method of claim 50, wherein said second detection means comprises a detectable moiety.