Broadly cross-reactive neutralizing antibodies against human immunodeficiency virus selected by env-cd4-co-receptor complexes

• US 20040259075A1
• Filed: 05/05/2004
• Published: 12/23/2004
• Est. Priority Date: 10/16/2001
• Status: Active Grant

• Current Assignee: The Scripps Research Institute, The United States of America As Represented By The Secretary of Agriculture
• Original Assignee: The United States of America As Represented By The Secretary of Agriculture
• Inventors: Shu, Yuuei, Dimitrov, Dimiter S., Phogat, Sanjay K., Zhang, Mei-Yun, Moulard, Maxime, Xiao, Xiadong, Burton, Dennis
• Granted Patent: US 7,223,844 B2
• Time in Patent Office: Days
• US Class Current: 435/5
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 31/14 for RNA viruses
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: C07K 14/70514 CD4
• CPC Class Codes: C07K 16/1063 env, e.g. gp41, gp110/120, gp160, V3, PND, CD4 binding site
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/55 Fab or Fab'
• CPC Class Codes: C07K 2319/00 Fusion polypeptide

Abstract

The present invention features antibodies and antibody fragments that specifically bind a CD4-inducible HIV gp120 epitope that is enhanced by binding a co-receptor for HIV, such as CCR5 or CXCR4, and pharmaceutical compositions comprising the antibodies or antibody fragments. The invention also features nucleic acids encoding the antibodies or antibody fragments, pharmaceutical compositions comprising the nucleic acids encoding the antibodies or antibody fragments, vectors comprising the nucleic acids, and cells comprising the vectors. The invention further features methods of identifying antibodies or antibody fragments with broadly neutralizing activity against HIV. The invention also features methods of inhibiting HIV entry into cells and methods of inhibiting replication of HIV in mammals, using the antibodies and nucleic acids of the invention.

Claims

1. An isolated antibody or antibody fragment that specifically binds a CD4-inducible epitope on Human Immunodeficiency Virus (HIV) Env, wherein exposure of the CD4-inducible epitope is further enhanced by the binding of Env to a co-receptor for HIV, wherein the CD4-inducible epitope is distinct from the HIV co-receptor binding site on gp120.

2. The isolated antibody or antibody fragment of claim 1, wherein the antibody or antibody fragment is selected by virtue of its ability to specifically bind to a CD4-inducible epitope on HIV Env that is enhanced by binding a co-receptor for HIV.

4. The isolated antibody or antibody fragment of claim 1, wherein the epitope is on gp120.

5. The isolated antibody or antibody fragment of claim 1, wherein the epitope is on gp41.

6. The isolated antibody or antibody fragment of claim 1, wherein the eipitope is on gp120-gp41.

9. The isolated antibody or antibody fragment of claim 1, wherein the HIV co-receptor is CCR5 or CXCR4.

10. The isolated antibody or antibody fragment of claim 1, wherein the HIV gp120 epitope is an HIV-1 gp120 epitope.

11. The isolated antibody or antibody fragment of claim 1, wherein the isolated antibody or antibody fragment has broadly neutralizing activity against HIV-1.

12. The isolated antibody or antibody fragment of claim 1, wherein the isolated antibody or antibody fragment is monoclonal.

13. The isolated antibody or active fragment thereof of claim 1, wherein the isolated antibody or antibody fragment is human or humanized.

14. The isolated antibody or antibody fragment of claim 1, wherein the isolated antibody or antibody fragment is isolated from a phage display library.

15. The isolated antibody or antibody fragment of claim 1, wherein the antibody or antibody fragment comprises the heavy chain of antibody Fab fragment X5 (SEQ ID NO:

3).

16. The isolated antibody or antibody fragment of claim 1, wherein the antibody or antibody fragment comprises the light chain of antibody Fab fragment X5 (SEQ ID NO:

2).

17. The isolated antibody or antibody fragment of claim 1, wherein the antibody or antibody fragment comprises the heavy chain of antibody Fab fragment X5 (SEQ ID NO:

3) and the light chain of antibody Fab fragment X5 (SEQ ID NO;

     2).

18. The isolated antibody or antibody fragment of claim 1, wherein the isolated antibody or antibody fragment comprises the CDR3 region (SEQ ID NO:

5) of the heavy chain of antibody Fab fragment X5.

19. The isolated antibody or antibody fragment of claim 1, wherein the isolated antibody or antibody fragment comprises the CDR3 region (SEQ ID NO:

8) of the light chain of antibody Fab fragment X5.

20. The isolated antibody or antibody fragment of claim 1, wherein a polypeptide comprising a heavy chain or light chain of the antibody or antibody fragment comprises soluble CD4.

21. The isolated antibody or antibody fragment of claim 20, wherein a polypeptide comprising the heavy chain or light chain of the antibody or antibody fragment further comprises peptide T20.

34. A pharmaceutical composition comprising the isolated antibody or antibody fragment of claim 1, and a pharmaceutically acceptable carrier.

35. The pharmaceutical composition of claim 34, further comprising soluble CD4.

36. A pharmaceutical composition comprising a nucleic acid that encodes the isolated antibody or antibody fragment of claim 1, and a pharmaceutically acceptable carrier.

37. The pharmaceutical composition of claim 36, wherein the nucleic acid is within an expression vector.

39. The method of claim 37, wherein the antibody is exposed to a complex comprising HIV gp120, CD4, and a co-receptor for HIV.

40. An antibody produced by the method of claim 39.

3. An isolated antibody or antibody fragment, wherein the isolated antibody or antibody fragment is selected by virtue of its ability to specifically bind to a CD4-inducible epitope on Human Immunodeficiency Virus (HIV) Env that is enhanced by the binding of Env to a co-receptor for HIV, wherein the CD4-inducible epitope is distinct from the HIV co-receptor binding site on gp120.

7. An isolated antibody or antibody fragment, wherein the isolated antibody or antibody fragment is selected by virtue of its ability to specifically bind to a complex comprising HIV gp120, CD4, and a co-receptor for HIV.

8. The isolated antibody or antibody fragment of claim 7, wherein the complex further comprises gp41.

22. An isolated polypeptide comprising the heavy chain of antibody Fab fragment X5 (SEQ ID NO:

3).

23. An isolated polypeptide comprising the light chain of antibody Fab fragment X5 (SEQ ID NO:

2).

24. An isolated polypeptide comprising the CDR3 region (SEQ ID NO:

5) of the heavy chain of antibody Fab fragment X5.

25. An isolated polypeptide comprising the CDR3 region (SEQ ID NO:

8) of the light chain of antibody Fab fragment X5.

26. An isolated nucleic acid that encodes SEQ ID NO:

3.

32. A vector comprising the isolated nucleic acid of claim 26.

33. A cell comprising the vector of claim 32.

27. An isolated nucleic acid that encodes SEQ ID NO:

2.

28. An isolated nucleic acid that encodes SEQ ID NO:

5.

29. An isolated nucleic acid that encodes SEQ ID NO:

8.

30. An isolated nucleic acid that encodes an antibody or antibody fragment comprising the heavy chain of antibody Fab fragment X5 (SEQ ID NO:

3) and the light chain of antibody Fab fragment X5 (SEQ ID NO;

     2).

31. The isolated nucleic acid of claim 30, wherein the isolated nucleic acid comprises the nucleotide sequence set forth in SEQ ID NO:

4.

38. A method of selecting an antibody or antibody fragment with broadly neutralizing activity against HIV, comprising detecting an antibody or antibody fragment that specifically binds a CD4-inducible epitope on HIV Env that is enhanced by the binding of Env to a co-receptor for HIV, wherein the CD4-inducible epitope is distinct from the HIV co-receptor binding site on gp120.

52. The method of claim 38, wherein the co-receptor is CCR5.

53. The method of claim 38, wherein the co-receptor is CXCR4.

41. A method of inhibiting entry of HIV into a cell, comprising administering to the cell an effective amount of an isolated antibody or antibody fragment that specifically binds a CD4-inducible epitope on HIV Env that is enhanced by the binding of Env to a co-receptor for HIV, wherein the CD4-inducible epitope is distinct from the HIV co-receptor binding site on gp120, thereby inhibiting entry of HIV into the cell.

42. The method of claim 41, wherein the cell is in a mammal that is susceptible to infection by HIV and wherein the isolated antibody or antibody fragment is administered to the mammal.

43. The method of claim 41, wherein the isolated antibody or antibody fragment is administered to the mammal by administering a nucleic acid encoding the isolated antibody or antibody fragment to the mammal.

44. The method of claim 42, wherein the mammal is a primate.

45. The method of claim 44, wherein the primate is a human.

46. The method of claim 41, wherein the HIV is HIV-1.

47. A method of inhibiting replication of HIV in a mammal that is susceptible to HIV infection, comprising administering to the mammal an effective amount of an isolated antibody or antibody fragment that specifically binds a CD4-inducible epitope on HIV Env that is enhanced by the binding of Env to a co-receptor for HIV, wherein the CD4-inducible epitope is distinct from the HIV co-receptor binding site on gp120, thereby inhibiting replication of HIV in the mammal.

48. The method of claim 47, wherein the isolated antibody or antibody fragment is administered to the mammal by administering a nucleic acid encoding the isolated antibody or antibody fragment to the mammal.

49. The method of claim 47, wherein the mammal is a primate.

50. The method of claim 49, wherein the primate is a human.

51. The method of claim 47, wherein the HIV is HIV-1.