DR5 antibodies and uses thereof

• US 20060269554A1
• Filed: 01/30/2006
• Published: 11/30/2006
• Est. Priority Date: 02/02/2005
• Status: Active Grant

• Current Assignee: Genentech, Inc. (Roche Holding AG)
• Original Assignee: Genentech, Inc. (Roche Holding AG)
• Inventors: Adams, Camellia W.
• Granted Patent: US 8,029,783 B2
• Time in Patent Office: Days
• US Class Current: 424/155.100
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 35/04 specific for metastasis
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 16/2878 against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]

Abstract

The invention concerns anti-DR5 antibodies with improved properties, compositions comprising such antibodies, methods and means for making such antibodies, and their therapeutic use, in particular in the treatment of cancer.

Claims

1. An anti-DR5 antibody comprising at least one mutation in the heavy and/or light chain of full-length antibody 16E2 (SEQ ID NOS:

11 and 13, respectively), or a fragment thereof, wherein said antibody or antibody fragment has a greater affinity for DR5 than said full-length antibody 16E2, and/or exhibits a greater biological activity and/or potency than said full-length antibody 16E2.

2. The anti-DR5 antibody of claim 1 wherein said antibody binds essentially to the same epitope as full-length antibody 16E2, or a fragment thereof.

3. The anti-DR5 antibody of claim 1 comprising a heavy and/or light chain having at least one substitution listed in any of Tables 1 through 7, and 9-12, or a fragment thereof.

4. The anti-DR5 antibody of claim 1 comprising one or more mutations in the framework of the full length 16E2 antibody heavy chain (SEQ ID NO:

11) variable domain, or a fragment thereof.

5. The anti-DR5 antibody of claim 4 wherein the framework mutation is selected from the group consisting of Q6E, V11L, E12V, R13Q, and K105Q, or a fragment thereof.

6. The anti-DR5 antibody of claim 5 comprising all of the framework mutations Q6E, V11L, E12V, R13Q, and K105Q, or a fragment thereof.

7. The anti-DR5 antibody of claim 1 or claim 6 comprising at least one mutation selected from the group consisting of T28A, G33A, M34L, M34A, M34I, M34S, N53Q, N53Y, and L102Y, in the amino acid sequence of SEQ ID NO:

11, or a fragment thereof.

8. The anti-DR5 antibody of claim 1 or claim 6 comprising at least one of the mutations G99A and R100A in the amino acid sequence of SEQ ID NO:

11, or a fragment thereof.

9. The anti-DR5 antibody of claim 6 comprising a set of mutations selected from the group consisting of (i) N53Q, L102Y;

(ii) M34L, N53Q, L102Y;

(iii) N53Y, L102Y;

(iv) M34L, N53Y, L102Y;

(v) G33A, N53Q, L102Y;

(vi) M34L, N53Y, L102Y;

(vii) G33A, N53Q, L102Y;

(viii) G33A, N53Y, L102Y;

(ix) T28A, N53Q, L102Y; and

(x) T28A, N53Y, L102Y in the amino acid sequence of SEQ ID NO;

11, or a fragment thereof.

10. The anti-DR5 antibody of claim 1 or claim 6, comprising at least one mutation in the light chain of full-length 16E2 antibody (SEQ ID NO:

13), or a fragment thereof.

11. The anti-DR5 antibody of claim 10 wherein the light chain is a lambda chain, or a fragment thereof.

12. The anti-DR5 antibody of claim 11 wherein said mutation is in CDR L1, or a fragment thereof.

13. The anti-DR5 antibody of claim 12 wherein said mutation is selected from the group consisting of Q24A, Q24S, G25A, D26E, S27A, L28A, R29A, S30A, Y31A, Y31K, Y32H, A33G, S34A, and S34Y in the amino acid sequence of SEQ ID NO:

13, or a fragment thereof.

14. The anti-DR5 antibody of claim 12 comprising a set of mutations selected from the group consisting of (i) Q24S, D26E, Y31K, S34Y;

and (ii) D26E, Y31K in the amino acid sequence of SEQ ID NO;

13, or a fragment thereof.

15. The anti-DR5 antibody of claim 11 wherein said mutation is in CDR L2, or a fragment thereof.

16. The anti-DR5 antibody of claim 15 wherein said mutation is selected from the group consisting of G50A, G50K, G50S, K51D, N52A, N52S, N52L, N52Q, N53A, N53E, N53Q, N53S, P55A, and S56A in the amino acid sequence of SEQ ID NO:

13.

17. The anti-DR5 antibody of claim 15 comprising a set of mutations selected from the group consisting of (i) G50K, K52S, N53E;

(ii) G50S, K51D, N52S, N53E;

(ii) N52S, N53E; and

(iv) N52Q, N53S, in the amino acid sequence of SEQ ID NO;

13, or a fragment thereof.

18. The anti-DR5 antibody of claim 11 wherein said mutation in CDR L3, or a fragment thereof.

19. The anti-DR5 antibody of claim 18 comprising at least one mutation selected from the group consisting of, N89A, N89L, N89Q, R91A, S93A, N95aA, N95aT, N95aQ, H95bA, N95bY, V96A, V97A in SEQ ID NO:

13, or a fragment thereof.

20. The anti-DR5 antibody of claim 18 comprising a set of mutations selected from the group consisting of (i) N89L, R91A, N95aT, H95bY;

and(ii) N95aT, H95bY in the sequence of SEQ ID NO;

13, or a fragment thereof.

21. The anti-DR5 antibody of claim 1 comprising a set of light chain mutations selected from the group consisting of (i) Q24S, G50K, K51D, H95bY;

(ii) Q24S, K51A, D92S, S93Y; and

(iii) Q24S, K51A, R91A in the amino acid sequence of SEQ ID NO;

13, or a fragment thereof.

22. The anti-DR5 antibody of claim 21 additionally comprising a set of heavy chain mutations selected from the group consisting of (ii) M34L, N53Q, L102Y;

(ii) M34L, N53Y, L102Y;

(iii) G33A, N53Q, L102Y;

(iv) G33A, N53Y, L102Y;

(v) M34L, N53Q, L102Y;

(vi) M34L, N53Y, L103Y;

(vii) G33A, N53Q, L102Y;

(viii) G33A, N53Y, L102Y; and

(ix) T28A, N53Q, L102Y in the amino acid sequence of SEQ ID NO;

11, or a fragment thereof.

23. The anti-DR5 antibody of claim 22 additionally comprising a set of framework mutations listed in Table 5, or a fragment thereof.

24. The anti-DR5 antibody of claim 1 comprising the following mutations:

G33A, N53Q, L102Y in the sequence of SEQ ID NO;

11; and

Q24S, K51A, R91A in the sequence of SEQ ID NO;

13, or a fragment thereof.

25. The anti-DR5 antibody of claim 24 additionally comprising at least one framework mutation, or a fragment thereof.

26. The anti-DR5 antibody of claim 25 wherein said framework mutation is at at least one of residues 6, 11, 12, 13, and 105 of SEQ ID NO:

11, or a fragment thereof.

27. The anti-DR5 antibody of claim 24 selected from the group consisting of Apomabs 1.1, 2.1, 3.1, 4.1, 5.1, 6.1, 7.1, 8.1, 9.1, 1.2, 2.2, 3.2, 4.2, 5.2, 6.2, 7.2, 8.2, 9.2, 1.3, 2.2, 3.3, 4.3, 5.3, 6.3, 7.3, 8.3, and 9.3, or a fragment thereof.

28. The anti-DR5 antibody of claim 27 which is Apomab 7.3 or 8.3, or a fragment thereof.

29. The anti-DR5 antibody of claim 28 which is Apomab 7.3, or a fragment thereof.

30. The anti-DR5 antibody of claim 1 wherein said antibody binds essentially to the same epitope as Apomab 7.3, or a fragment thereof.

31. The anti-DR5 antibody of claim 1 or claim 29, wherein said fragment is selected from the group consisting of Fab, Fab′

, F(ab′

)2, and Fv fragments, diabodies, single chain antibody molecules, and multispecific antibodies formed from antibody fragments.

32. The anti-DR5 antibody of claim 31 wherein said antibody is a single-chain antibody.

33. The anti-DR5 antibody of claim 31 wherein said fragment is an Fv fragment.

34. The anti-DR5 antibody or antibody fragment of claim 1 wherein said biological activity is anti-cancer activity.

35. The anti-DR5 antibody or antibody fragment of claim 1 wherein said biological activity is activation or stimulation of apoptosis in cancer cells.

36. The anti-DR5 antibody or antibody fragment of claim 34 wherein cancer is selected from the group consisting of carcinoma, lymphoma, blastoma, sarcoma and leukemia.

37. The anti-DR5 antibody or antibody fragment of claim 36 wherein said cancer is selected from the group consisting of squamous cell cancer, small-cell lung cancer, non-small cell lung cancer (NSCLC), non-Hodgkin'"'"'s lymphoma, blastoma, gastrointestinal cancer, renal cancer, ovarian cancer, liver cancer, stomach cancer, bladder cancer, hepatoma, breast cancer, colon cancer, colorectal cancer, pancreatic cancer endometrial carcinoma, salivary gland carcinoma, kidney cancer, liver cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma, and head and neck cancer.

38. The anti-DR5 antibody or antibody fragment of claim 37 wherein said cancer is NSCLC, non-Hodgkin'"'"'s lymphoma, colorectal cancer, or pancreatic cancer.

39. The anti-DR5 antibody or antibody fragment of claim 36 wherein said cancer is an adenocarcinoma.

40. The anti-DR5 antibody or antibody fragment of claim 39 wherein said adenocarcinoma is colorectal, pancreatic, or metastatic adenocarcinoma.

41. The DR5 antibody or antibody fragment of claim 1 wherein said potency is determined in an in vitro tumor killing assay.

42. The anti-DR5 antibody or antibody fragment of claim 1 which is a chimeric, humanized, or human antibody.

43. The anti-DR5 antibody or antibody fragment of claim 1 which mediates antibody-dependent cellular cytotoxicity (ADCC).

44. The anti-DR5 antibody of claim 1 which comprises Apomab 7.3, or Apomab 8.3, or a fragment thereof.

45. The anti-DR5 antibody of claim 1 in a dimeric form, or a fragment thereof.

46. The anti-DR5 antibody of claim 2 cross-linked with an anti-human IgG Fc region, or a fragment thereof.

47. The anti-DR5 antibody or antibody fragment of claim 1 fused to an epitope tag sequence.

48. A chimeric molecule comprising the antibody of claim 44 fused to a heterologous amino acid sequence.

49. The chimeric molecule of claim 48 wherein said heterologous amino acid sequence comprises an immunoglobulin sequence.

50. The chimeric molecule of claim 49 wherein said immunoglobulin sequence is an anti-human IgG Fc region.

51. Isolated nucleic acid encoding the heavy or light chain of an anti-DR5 antibody of claim 1.

53. A vector comprising the nucleic acid of claim 51 or claim 52.

54. A host cell comprising the nucleic acid of claim 51 or claim 52.

55. A method of producing an anti-DR5 antibody comprising culturing the host call of claim 54 under conditions wherein the DNA is expressed.

56. A composition comprising an anti-DR5 antibody of claim 1, 28 or 29, or a fragment thereof, and a carrier.

57. The composition of claim 56 wherein said carrier is a pharmaceutically-acceptable carrier.

58. The composition of claim 57 further comprising an additional anti-cancer agent.

59. The composition of claim 58 wherein said additional anti-cancer agent is an antibody.

60. The composition of claim 59 wherein said antibody is selected from the group consisting of an additional anti-DR5 antibody, Rituxan (rituximab), and an anti-VEGF antibody.

61. The composition of claim 58 wherein said additional anti-cancer agent is a chemotherapeutic agent.

62. The composition of claim 61 wherein said chemotherapeutic agent is selected from the group consisting of CPT-11 (irinotecan), gemcitabine, carboplatin, taxol, and paclitaxel.

63. The composition of claim 58 wherein said additional anti-cancer agent is an Apo2L ligand comprising amino acids 114-281 of FIG. 1 (SEQ ID NO:

1).

64. A method of inducing apoptosis comprising exposing mammalian cancer cells to an anti-DR5 antibody of claim 1, claim 28, or claim 29, or a fragment thereof.

65. The method of claim 64 wherein said antibody is a chimeric, humanized or human antibody.

66. The method of claim 64 wherein said mammalian cancer cells are exposed to an agent which activates DR5.

67. A method for the treatment of cancer comprising administering to a mammalian subject an effective amount of an anti-DR5 antibody of claim 1, 28, or 29, or a fragment thereof.

68. The method of claim 67 wherein said mammalian subject is a human patient.

69. The method of claim 68 wherein said cancer is selected from the group consisting of squamous cell cancer, small-cell lung cancer, non-small cell lung cancer (NSCLC), non-Hodgkin'"'"'s lymphoma, blastoma, gastrointestinal cancer, renal cancer, ovarian cancer, liver cancer, stomach cancer, bladder cancer, hepatoma, breast cancer, colon cancer, colorectal cancer, pancreatic cancer, endometrial carcinoma, salivary gland carcinoma, kidney cancer, liver cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma, and head and neck cancer.

70. The method of claim 69 wherein said cancer is NSCLC, colorectal cancer, non-Hodgkin'"'"'s lymphoma or pancreatic cancer.

71. The method of claim 68 wherein said cancer is an adenocarcinoma.

72. The method of claim 71 wherein said adenocarcinoma is colorectal, pancreatic, or metastatic adenocarcinoma.

73. The method of any one of claims 67-72, additionally comprising the administration of a further anticancer agent.

74. An article of manufacture comprising a container and compositions contained within said container, wherein the composition includes an anti-DR5 antibody of claim 1, 28, or 29, or a fragment thereof.

75. The article of manufacture of claim 74 further comprising instructions for using the anti-DR5 antibody in vitro or in vivo.

76. The article of manufacture of claim 75 wherein said instructions concern the treatment of cancer.

52. Isolated nucleic acid encoding the heavy or light chain of Apomab 7.3 or Apomab 8.3.