Antibodies that immunospecifically bind to trail receptors

• US 20030190685A1
• Filed: 05/07/2002
• Published: 10/09/2003
• Est. Priority Date: 05/25/2001
• Status: Active Grant

• Current Assignee: Human Genome Sciences Incorporated (GlaxoSmithKline PLC)
• Original Assignee: Human Genome Sciences Incorporated (GlaxoSmithKline PLC)
• Inventors: Vaughan, Tristan, Rosen, Craig A., Ruben, Steven M., Albert, Vivian R., Dobson, Claire, Salcedo, Theodora
• Granted Patent: US 7,064,189 B2
• Time in Patent Office: Days
• US Class Current: 435/7.23
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/00 Medicinal preparations containing organic active ingredients
• CPC Class Codes: A61K 39/395 Antibodies agglutinins A61K38/36 ; as drug carriers A61K47/50; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/18 for pancreatic disorders, e.g. pancreatic enzymes
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• 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 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)

Abstract

The present invention relates to antibodies and related molecules that immunospecifically bind to TRAIL receptor, TR4. Such antibodies have uses, for example, in the prevention and treatment of cancers and other proliferative disorders. The invention also relates to nucleic acid molecules encoding anti-TR4 antibodies, vectors and host cells containing these nucleic acids, and methods for producing the same. The present invention relates to methods and compositions for preventing, detecting, diagnosing, treating or ameliorating a disease or disorder, especially cancer and other hyperproliferative disorders, comprising administering to an animal, preferably a human, an effective amount of one or more antibodies or fragments or variants thereof, or related molecules, that immunospecifically bind to TRAIL receptor TR4.

Claims

1. An isolated antibody or fragment thereof comprising a first amino acid sequence at least 95% identical to a second amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of either VHCDR1, VHCDR2, or VHCDR3 of any one of SEQ ID NOS;

42-53; and

(1) the amino acid sequence of either VLCDR1, VLCDR2, or VLCDR3 of any one of SEQ ID NOS;

42-53;

wherein said antibody or fragment thereof immunospecifically binds TR4.

2. The antibody or fragment thereof of claim 1, wherein the second amino acid sequence consists of the amino acid sequence of a VHCDR3 of any one of SEQ ID NOS:

42-53.

3. The antibody or fragment thereof of claim 1, that preferentially binds TR4 relative to its ability to bind TR1, TR5, TR7 and TR10.

4. The antibody or fragment thereof of claim 1, that binds TR4 expressed on the surface of a cell.

5. An isolated antibody or fragment thereof comprising:

(a) an amino acid sequence that is at least 90% identical to a VH domain of any one of SEQ ID NOS;

42-53;

(b) an amino acid sequence that is at least 90% identical to a VL domain of any one of SEQ ID NOS;

42-53;

or

(c) both (a) and (b);

wherein said antibody or fragment thereof immunospecifically binds TR4.

6. The antibody or fragment thereof of claim 5, wherein the VH domain has the amino acid sequence of the VH domain of SEQ ID NO:

42 and the VL domain has the amino acid sequence of the VL domain of SEQ ID NO;

42.

7. The antibody or fragment thereof of claim 5, wherein the VH domain has the amino acid sequence of the VH domain of SEQ ID NO:

43 and the VL domain has the amino acid sequence of the VL domain of SEQ ID NO;

43.

8. The antibody or fragment thereof of claim 5, wherein the VH domain has the amino acid sequence of the VH domain of SEQ ID NO:

48 and the VL domain has the amino acid sequence of the VL domain of SEQ ID NO;

48.

9. The antibody or fragment thereof of claim 5, that preferentially binds TR4 relative to its ability to bind TR1, TR5, TR7 and TR10.

10. The antibody or fragment thereof of claim 5, that binds TR4 expressed on the surface of a cell.

11. The antibody or fragment thereof of claim 5 comprising:

(a) the amino acid sequence of a VH domain of any one of SEQ ID NOS;

42-53;

(b) the amino acid sequence of a VL domain of any one of SEQ ID NOS;

42-53;

or

(c) both (a) and (b);

wherein said antibody or fragment thereof immunospecifically binds TR4.

12. The antibody or fragment thereof of claim 11, wherein the VH domain has the amino acid sequence of the VI domain SEQ ID NO:

42 and the VL domain has the amino acid sequence of the VL domain of SEQ ID NO;

42.

13. The antibody or fragment thereof of claim 11, wherein the VH domain has the amino acid sequence of the VH domain SEQ ID NO:

43 and the VL domain has the amino acid sequence of the VL domain of SEQ ID NO;

43.

14. The antibody or fragment thereof of claim 11, wherein the VH domain has the amino acid sequence of the VH domain SEQ ID NO:

48 and the VL domain has the amino acid sequence of the VL domain of SEQ ID NO;

48.

15. The antibody or fragment thereof of claim 11, that preferentially binds TR4 relative to its ability to bind TR1, TR5, TR7 and TR10.

16. The antibody or fragment thereof of claim 11, that binds TR4 expressed on the surface of a cell.

17. The antibody or fragment thereof of claim 5 wherein the antibody or fragment thereof is selected from the group consisting of:

(a) a whole immunoglobulin molecule;

(b) an scFv;

(c) a monoclonal antibody;

(d) a human antibody;

(e) a chimeric antibody;

(f) a humanized antibody;

(g) a Fab fragment;

(h) an Fab′

fragment;

(i) an F(ab′

)2;

(j) an Fv; and

(k) a disulfide linked Fv.

18. The antibody or fragment thereof of claim 5 which comprises a heavy chain immunoglobulin constant domain selected from the group consisting of:

(a) a human IgM constant domain;

(b) a human IgG1 constant domain;

(c) a human IgG2 constant domain;

(d) a human IgG3 constant domain;

(e) a human IgG4 constant domain; and

(f) a human IgA constant domain.

19. The antibody or fragment thereof of claim 5 which comprises a light chain immunoglobulin constant domain selected from the group consisting of:

(a) a human Ig kappa constant domain; and

(b) a human Ig lambda constant domain.

20. The antibody or fragment thereof of claim 5 wherein the antibody or fragment thereof has a dissociation constant (K_D) selected from the group consisting of:

(a) a dissociation constant (K_D) between 10<sup>−

7</sup> M (inclusive) and 10<sup>−

8</sup> M; and

(b) a dissociation constant (K_D) between 10<sup>−

8</sup> M (inclusive) and 10<sup>−

9</sup> M.

21. The antibody or fragment thereof of claim 5 wherein the antibody or fragment thereof has a dissociation constant (K_D) of less than or equal to 10⁻

9 M.

22. The antibody or fragment thereof of claim 21 wherein the antibody or fragment thereof has a K_D between 10⁻

9 M and 10<sup>−

10</sup> M.

23. The antibody or fragment thereof of claim 21 wherein the antibody or fragment thereof has a K_D between 10⁻

10 M (inclusive) and 10<sup>−

11</sup> M.

24. The antibody or fragment thereof of claim 21 wherein the antibody or fragment thereof has a K_D between 10⁻

11 M (inclusive) and 10<sup>−

12</sup> M.

25. The antibody or fragment thereof of claim 5 wherein the antibody or fragment thereof is conjugated to a detectable label.

26. The antibody or fragment thereof of claim 25, wherein the detectable label is a radiolabel.

27. The antibody or fragment thereof of claim 26, wherein the radiolabel is ¹²⁵I, ¹³¹I, ¹¹¹In, ⁹⁰Y, ⁹⁹Tc, ¹⁷⁷Lu, ¹⁶⁶Ho, or ¹⁵³Sm.

28. The antibody or fragment thereof of claim 25, wherein the detectable label is an enzyme, a fluorescent label, a luminescent label, or a bioluminescent label.

29. The antibody or fragment thereof of claim 5 wherein the antibody or fragment thereof is biotinylated.

30. The antibody or fragment thereof of claim 5 wherein the antibody or fragment thereof is conjugated to a therapeutic or cytotoxic agent.

31. The antibody or fragment thereof of claim 30, wherein the therapeutic or cytotoxic agent is selected from the group consisting of:

(a) an anti-metabolite;

(b) an alkylating agent;

(c) an antibiotic;

(d) a growth factor;

(e) a cytokine;

(f) an anti-angiogenic agent;

(g) an anti-mitotic agent;

(h) an anthracycline;

(i) toxin; and

(j) an apoptotic agent.

32. The antibody or fragment thereof of any one of claim 5 wherein the antibody or fragment thereof is attached to a solid support.

33. The antibody or fragment thereof of claim 5 wherein the antibody or fragment thereof immunospecifically binds TR4 in a Western blot.

34. The antibody or fragment thereof of claim 5 wherein the antibody or fragment thereof imimunospecifically binds TR4 in an ELISA.

35. An isolated cell that produces the antibody or fragment thereof of claim 11.

36. The antibody or fragment thereof of claim 5 that does not inhibit the ability of TRAIL to bind TR4.

37. The antibody or fragment thereof of claim 5 wherein the antibody or fragment thereof is an agonist of TR4.

38. The antibody or fragment thereof of claim 5 wherein the antibody or fragment thereof stimulates apoptosis of TR4 expressing cells.

39. The antibody or fragment thereof of claim 38 wherein the antibody or fragment thereof stimulates apoptosis of TR4 expressing cells better than an equal concentration of TRAIL polypeptide stimulates apoptosis of TR4 expressing cells.

40. The antibody or fragment thereof of claim 38 wherein the antibody or fragment thereof stimulates apoptosis of TR4 expressing cells equally well in the presence or absence of antibody cross-linking reagents.

41. The antibody or fragment thereof of claim 38 wherein the antibody or fragment thereof is not hepatotoxic.

42. The antibody or fragment thereof of claim 5 wherein the antibody or fragment thereof upregulates TRAIL receptor expression.

43. The antibody or fragment thereof of claim 5 wherein the antibody or fragment thereof inhibits TRAIL binding to TR4.

44. The antibody or fragment thereof of claim 5 wherein the antibody or fragment thereof is an antagonist of TR4.

45. The antibody or fragment thereof of claim 5 wherein the antibody or fragment thereof inhibits apoptosis of TR4 expressing cells.

46. The antibody or fragment thereof of claim 5 wherein the antibody or fragment thereof downregulates TRAIL receptor expression.

47. An antibody or fragment thereof that binds the same epitope on a TR4 polypeptide as an antibody or fragment thereof of claim 11.

48. An antibody or fragment thereof t hat b inds the s ame epitope on a TR4 polypeptide as an antibody or fragment thereof of claim 12.

49. An antibody or fragment thereof that binds the same epitope on a TR4 polypeptide as an antibody or fragment thereof of claim 13.

50. An antibody or fragment thereof that binds the same epitope on a TR4 polypeptide as an antibody or fragment thereof of claim 14.

51. The antibody or fragment thereof of claim 5 in a pharmaceutically acceptable carrier.

52. A method of treating, preventing or ameliorating a cancer comprising administering to an animal the antibody or fragment thereof of claim 5 or a composition containing said antibody or fragment thereof.

53. The method of claim 52 wherein the animal is a human.

54. The method of claim 52 wherein the cancer is colon cancer.

55. The method of claim 52 wherein the cancer is breast cancer.

56. The method of claim 52 wherein the cancer is uterine cancer.

57. The method of claim 52 wherein the cancer is pancreatic cancer.

58. The method of claim 52 wherein the cancer is lung cancer.

59. The method of claim 52 wherein the cancer is gastrointestinal cancer.

60. The method of claim 52 wherein the cancer is Kaposi'"'"'s sarcoma.

61. The method of claim 52 wherein the cancer is a cancer of the central nervous system.

62. The method of claim 61 wherein the cancer of the central nervous system is a medulloblastoma.

63. The method of claim 61 wherein the cancer of the central nervous system is a neuroblastoma.

64. The method of claim 61 wherein the cancer of the central nervous system is a glioblastoma.

65. The method of claim 52, wherein the antibody or fragment thereof is administered in combination with a chemotherapeutic agent.

66. The method of claim 65, wherein chemotherapeutic agent is selected from the group consisting of:

(a) irinotecan;

(b) paclitaxel (TAXOL)®

3; and

(c) gemcitabine.

67. A method of treating, preventing or ameliorating a disease or disorder selected from the group consisting of:

(a) graft vs. host disease (GVHD);

(b) AIDS; and

(c) a neurodegenerative disorder;

comprising administering to an animal the antibody or fragment thereof of claim 5 or a composition containing said antibody or portion theeof.

68. The method of claim 67, wherein the animal is a human.

69. A method of inhibiting the growth of or killing TR4 expressing cells, comprising administering to an animal in which such inhibition of growth or killing of TR4 receptor expressing cells is desired, the antibody or fragment thereof of claim 5 or a composition containing said antibody or fragment thereof in an amount effective to inhibit the growth of or kill TR4 expressing cells.

70. A method of detecting expression of a TR4 polypeptide comprising:

(a) assaying the expression of a TR4 polypeptide in a biological sample from an individual using the antibody or fragment thereof of claim 5;

and

(b) comparing the level of a TR4 polypeptide with a standard level of a TRAIL receptor polypeptide.

71. A method of detecting, diagnosing, prognosing, or monitoring cancers and other hyperproliferative disorders comprising:

(a) assaying the expression of a TR4 polypeptide in a biological sample from an individual using the antibody or fragment thereof of claim 5;

and

(b) comparing the level of a TR4 polypeptide with a standard level of TR4 polypeptide.

72. A kit comprising the antibody or fragment thereof of claim 5.

73. The kit of claim 72 comprising a control antibody.

74. The kit of claim 72, wherein the antibody or fragment thereof is coupled or conjugated to a detectable label.

75. The antibody expressed by the cell line of ATCC Deposit PTA-3570.

76. The antibody expressed by the cell line of ATCC Deposit PTA-3571.

77. The antibody expressed by the cell line of ATCC Deposit PTA-3675.