Humanized antibodies to LIV-1 and use of same to treat cancer

Patent metadata

Field	Value
Publication number	US-RE48959-E
Application number	US-201115862389-A
Country	US
Kind code	E1
Filing date	Dec 6, 2011
Priority date	Dec 6, 2010
Publication date	Mar 8, 2022
Grant date	Mar 8, 2022

Abstract

The invention provides humanized antibodies that specifically bind to LIV-1. The antibodies are useful for treatment and diagnoses of various cancers as well as detecting LIV-1.

First claim

What is claimed is:

1. A humanized antibody, that specifically binds human LIV-1 comprising a mature heavy chain variable region comprising three CDRs of SEQ ID NO:53 and having an amino acid sequence at least 95% identical to SEQ ID NO:53 provided that position H27 is occupied by L, position H29 is occupied by I, H30 by E and H94 by V and a mature light chain variable region comprising three CDRs of SEQ ID NO:60 at least 95% identical to SEQ ID NO:60 provided position L36 is occupied by Y and position L46 by P.

2. The humanized antibody of claim 1 , further provided position H76 is occupied by N.

3. The humanized antibody of claim 1 wherein the mature heavy chain variable region is fused to a heavy chain constant region and the mature light chain variable region is fused to a light chain constant region.

4. The humanized antibody of claim 3 , wherein the heavy chain constant region is a mutant form of a natural human constant region which has reduced binding to an Fcgamma receptor relative to the natural human constant region.

5. The humanized antibody of claim 3 , wherein the heavy chain constant region is of IgG1 isotope.

6. The humanized antibody of claim 3 , wherein the heavy chain constant region has an amino acid sequence comprising SEQ ID NO:44 and the light chain constant region has an amino acid sequence comprising SEQ ID NO:42.

7. The humanized antibody of claim 3 , wherein the heavy chain constant region has an amino acid sequence comprising SEQ ID NO:46 (S239C) and the light chain constant region has an amino acid sequence comprising SEQ ID NO:42.

8. The humanized antibody of claim 1 , wherein the mature heavy chain variable region has an amino acid sequence designated SEQ ID NO:52 or 53 and the mature light chain variable region has an amino acid sequence designated SEQ ID NO: 59 or 60.

9. The humanized antibody of claim 1 , wherein the mature heavy chain variable region has an amino acid sequence designated SEQ ID NO:53 and the mature light chain variable region has an amino acid sequence designated SEQ ID NO:60.

10. The humanized antibody of claim 1 , wherein the antibody is conjugated to a cytotoxic or cytostatic agent.

11. The humanized antibody of claim 1 , having an association constant for human or cynomolgus monkey LIV-1 of 0.5 to 2×10 9 M −1 .

12. A nucleic acid encoding a the mature heavy chain variable region and/or a the mature light chain variable region as defined by claim 1 .

13. A method of treating a patient having a cancer expressing LIV-1, comprising administering to the patient an effective regime of a the humanized antibody of claim 1 .

14. The method of claim 13 , wherein the cancer is a breast cancer, a prostate cancer, a cervical cancer or a melanoma.

15. A pharmaceutical composition comprising a the humanized antibody of claim 1 .

16. The method of claim 13 , wherein the antibody is conjugated to a cytotoxic or cytostatic agent.

17. The method of claim 13 , wherein the isotype of the antibody is human IgG1.

18. A vector comprising the nucleic acid of claim 12.

19. The vector of claim 18, wherein the vector is an expression vector.

20. A host cell comprising the nucleic acid of claim 12.

21. The host cell of claim 20, wherein the host cell is a Chinese hamster ovary (CHO) cell.

22. A method of producing an anti-LIV-1 antibody comprising culturing the host cell of claim 20 under a condition suitable for production of the anti-LIV-1 antibody.

23. The method of claim 22, further comprising isolating the anti-LIV-1 antibody produced by the host cell.

24. A method of producing an anti-LIV-1 antibody-drug conjugate comprising culturing the host cell of claim 20 under a condition suitable for production of an anti-LIV-1 antibody; isolating the anti-LIV-1 antibody produced from the host cell; and conjugating the anti-LIV-1 antibody to a cytotoxic or cytostatic agent.

25. The method of claim 24, wherein the cytotoxic or cytostatic agent is an antitubulin agent.

26. The method of claim 24, wherein the cytotoxic or cytostatic agent is an auristatin.

27. The method of claim 26, wherein the auristatin is monomethyl auristatin E.

28. The method of claim 26, wherein the auristatin is monomethyl auristatin F.

29. The method of claim 24, wherein the anti-LIV-1 antibody is conjugated to the cytotoxic or cytostatic agent via a linker.

30. The method of claim 29, wherein the linker is a cleavable peptide linker.

31. The method of claim 30, wherein the cleavable peptide linker has a formula:
-A a -W w —Y y —,
wherein: -A- is a stretcher unit; a is 0 or 1; each —W— is independently an amino acid unit; w is independently an integer ranging from 0 to 12; −Y— is a spacer unit; and y is 0, 1 or 2.

32. The method of claim 29, wherein the linker is attached to sulphydryl residues of the humanized antibody obtained by partial reduction or full reduction of the humanized antibody.

33. The method of claim 29, wherein the cytotoxic or cytostatic agent is monomethyl auristatin E.

34. The method of claim 33, wherein the anti-LIV-1 antibody-drug conjugate has the following structure:

wherein p denotes a number from 1 to 8 and Ab designates the anti-LIV-1 antibody.

35. The method of claim 34, wherein the average value of p in a population of the antibody-drug conjugate is about 4.

36. A nucleic acid encoding the mature heavy chain variable region and/or the mature light chain variable region as defined by claim 9.

37. A vector comprising the nucleic acid of claim 36.

38. The vector of claim 37, wherein the vector is an expression vector.

39. A host cell comprising the nucleic acid of claim 36.

40. The host cell of claim 39, wherein the host cell is a Chinese hamster ovary (CHO) cell.

41. A method of producing an anti-LIV-1 antibody comprising culturing the host cell of claim 39 under a condition suitable for production of the anti-LIV-1 antibody.

42. The method of claim 41, further comprising isolating the anti-LIV-1 antibody produced by the host cell.

43. A method of producing an anti-LIV-1 antibody-drug conjugate comprising culturing the host cell of claim 39 under a condition suitable for production of an anti-LIV-1 antibody; isolating the anti-LIV-1 antibody produced from the host cell; and conjugating the anti-LIV-1 antibody to a cytotoxic or cytostatic agent.

44. The method of claim 43, wherein the cytotoxic or cytostatic agent is an antitubulin agent.

45. The method of claim 43, wherein the cytotoxic or cytostatic agent is an auristatin.

46. The method of claim 45, wherein the auristatin is monomethyl auristatin E.

47. The method of claim 45, wherein the auristatin is monomethyl auristatin F.

48. The method of claim 43, wherein the anti-LIV-1 antibody is conjugated to the cytotoxic or cytostatic agent via a linker.

49. The method of claim 48, wherein the linker is a cleavable peptide linker.

Assignees

• Seagen Inc

Inventors

• Smith Maria Leia
• Sussman Django
• Arthur William
• Nesterova Albina

Classifications

• C07K2317/92

  Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value · CPC title

• C07K2317/732

  Antibody-dependent cellular cytotoxicity [ADCC] · CPC title

• C07K2317/565

  Complementarity determining region [CDR] · CPC title

• C07K2317/52

  Constant or Fc region; Isotype · CPC title

• C07K16/3069

  Reproductive system, e.g. ovaria, uterus, testes, prostate · CPC title