Antibodies that immunospecifically bind to TRAIL receptors

• US 20030228309A1
• Filed: 11/13/2002
• Published: 12/11/2003
• Est. Priority Date: 11/08/2000
• Status: Abandoned Application

• Current Assignee: Human Genome Sciences Incorporated (GlaxoSmithKline PLC)
• Original Assignee: Human Genome Sciences Incorporated (GlaxoSmithKline PLC)
• Inventors: Rosen, Craig A., Ruben, Steven M., Roschke, Viktor, Salcedo, Theodora
• Application Number:
  US10/292,486
• Publication Number:
  US 20030228309A1
• Time in Patent Office: Days
• US Class Current: 424/144.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: C07K 16/2878 against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto

Abstract

The present invention relates to antibodies and related molecules that immunospecifically bind to TRAIL receptors. 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-TRAIL receptor 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.

Claims

1. An isolated antibody that immunospecifcally binds TR4 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, 44, or 46; and

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

43, 45, or 47.

2. The antibody 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, 44, or 46.

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

6. The antibody of claim 1 wherein the antibody 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.

7. The antibody of claim 1 which also 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;

(e) a human IgG2 constant domain;

(d) a human IgG3 constant domain;

(e) a human IgG4 constant domain; and

(f) a human IgA constant domain.

8. The antibody of claim 1 which also comprises a light chain immunoglobulin constant domain selected from the group consisting of:

(a) a human Ig kappa constant domain;

(b) a human Ig lambda constant domain.

9. The antibody of claim 1 wherein the antibody is multimeric.

10. The antibody of claim 9 wherein the antibody is a dimer.

11. The antibody of claim 9 wherein the antibody is crosslinked.

12. An antibody that competitively inhibits the binding of the antibody of claim 1 to TR4.

13. The antibody of claim 1 wherein the antibody has a dissociation constant (K_D) selected from the group consisting of:

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

7</sup> M and 10<sup>−

8</sup> M;

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

8</sup> M and 10<sup>−

9</sup> M;

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

9</sup> M and 10<sup>−

10</sup> M;

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

10</sup> M and 10<sup>−

11</sup> M;

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

11</sup> M and 10<sup>−

12</sup> M; and

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

12</sup> M and 10<sup>−

13</sup> M.

14. The antibody of claim 1 wherein the antibody is associated with a detectable label.

15. The antibody of claim 14, wherein the detectable label is a radiolabel.

16. The antibody of claim 15, wherein the radiolabel is ¹²⁵I, ¹³¹I, ¹¹¹In, ⁹⁰Y, ⁹⁹Tc, ¹⁷⁷Lu, ¹⁶⁶Ho, ¹⁵³Sm, ²¹⁵Bi or ²²⁵Ac.

17. The antibody of claim 14, wherein the detectable label is an enzyme, a fluorescent label, a luminescent label, or a bioluminescent label.

18. The antibody of claim 1 wherein the antibody is biotinylated.

19. The antibody of claim 1 wherein the antibody is conjugated to a therapeutic or cytotoxic agent.

20. The antibody of claim 19, wherein the therapeutic or cytotoxic agent is selected from the group consisting of:

(a) an anti-metabolite,

(b) an alkylatingagent;

(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.

21. The antibody of any one of claim 1 which is attached to a solid support.

22. The antibody or portion thereof of claim 1 wherein said antibody immunospecifically binds to TR4 in a Western blot.

23. The antibody or portion thereof of claim 1 wherein said antibody immunospecifically binds to TR4 in an ELISA.

24. An isolated cell that produces the antibody of claim 1.

25. The antibody of claim 1 that diminishes or abolishes the ability of TRAIL to bind to TR4.

26. The antibody of claim 1 wherein the antibody is an agonist of a TR4 receptor.

27. The antibody of claim 1 wherein the antibody stimulates apoptosis of TR4 expressing cells.

28. The antibody of claim 27 wherein the antibody stimulates apoptosis of TR4 expressing cells better than an equal concentration of TRAIL polypeptide stimulates apoptosis of TR4 expressing cells.

29. The antibody of claim 27 wherein the antibody stimulates apoptosis of TR4 expressing cells equally well in the presence or absence of antibody cross-linking reagents.

30. The antibody of claim 27 which is not hepatotoxic.

31. The antibody of claim 1 wherein the antibody upregulates TRAIL receptor expression.

32. The antibody of claim 1 wherein the antibody inhibits TRAIL binding to a TR4.

33. The antibody of claim 1 wherein the antibody is an antagonist of TR4.

34. The antibody of claim 1 wherein the antibody inhibits apoptosis of TR4 expressing cells.

35. The antibody of claim 1 wherein the antibody downregulates TRAIL receptor expression.

36. An antibody that binds the same epitope on a TR4 polypeptide as an antibody of claim 1.

37. The antibody of claim 1 in a pharmaceutically acceptable carrier.

38. A method of treating, preventing or ameliorating a cancer comprising administering the antibody of claim 1 or a composition containing the antibody of claim 1 to an animal.

39. The method of claim 38, wherein the animal is a human.

40. The method of claim 38 wherein the cancer is selected from the group consisting of:

(a) colon cancer;

(b) breast cancer;

(c) uterine cancer;

(d) pancreatic cancer;

(e) lung cancer;

(f) gastrointestinal cancer; and

(g) Kaposi'"'"'s sarcoma.

41. The method of claim 38, wherein the antibody is administered in combination with a chemotherapeutic agent.

42. The method of claim 41, wherein chemotherapeutic agent is selected from the group consisting of:

(a) irinotecan;

(b) paclitaxel (TAXOL)®

; and

(c) gemcitabine.

43. The method of claim 38, wherein the antibody is multimeric.

44. The method of claim 38, wherein the antibody is a dimer.

45. The method of claim 38, wherein the antibody is adminstered in combination with a cross-linking agent.

46. The method of claim 38, wherein the antibody is administered in combination with an antibody that specifically binds TR7.

47. The method of claim 38, wherein the antibody is administered in combination with TRAIL and an antibody that specifically binds TR7.

48. The method of claim 38, wherein the antibody is administered in combination with TRAIL.

49. 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 of claim 1 or a composition containing the antibody of claim 1 in an amount effective to inhibit the growth of or kill TR4 expressing cells.

50. 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 of claim 1;

and

(b) comparing the level of a TR4 polypeptide with a standard level of a TRAIL receptor polypeptide, (e.g., the level in normal biological samples).

51. 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 of claim 1;

and

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

52. A kit comprising the antibody of claim 1.

53. The kit of claim 52 comprising a control antibody.

54. The kit of claim 52, wherein the antibody is coupled or conjugated to a detectable label.

4. An isolated antibody comprising:

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

42, 44, or 46;

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

43, 45, or 47;

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

wherein said antibody immunospecifically binds to TR4.

5. The antibody of claim 4 comprising:

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

42, 44, or 46;

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

43, 45, or 47;

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

wherein said antibody immunospecifically binds to TR4.

55. A hybridoma cell line selected from the hybridoma cell lines contained in ATCC Deposit Nos. PTA-3149, PTA-2687, PTA-3369, PTA-2730, PTA-2729, PTA-2728, PTA-3368, and PTA-2731.

56. An antibody that binds TR4 selected from the group consisting of

(a) the antibody expressed by the 7.1.3 cell line contained in ATCC Deposit No. PTA-3149;

(b) the antibody expressed by the 7.3.1 cell line contained in ATCC Deposit No. PTA-2687;

(c) the antibody expressed by the 7.3.2 cell line contained in ATCC Deposit No. PTA-2687;

(d) the antibody expressed by the 7.3.3 cell line contained in ATCC Deposit No. PTA-3369;

(e) the antibody expressed by the 7.8.1 ccell line contained in ATCC Deposit No. PTA-2730;

(b) the antibody expressed by the 7.8.2 cell line contained in ATCC Deposit No. PTA-2730;

(c) the antibody expressed by the 7.8.3 cell line contained in ATCC Deposit No. PTA-2730;

(f) the antibody expressed by the 7.10.1 ccell line contained in ATCC Deposit No. PTA-2729;

(b) the antibody expressed by the 7.10.2 cell line contained in ATCC Deposit No. PTA-2729;

(g) the antibody expressed by the 7.10.3 cell line contained in ATCC Deposit No. PTA-2729;

(h) the antibody expressed by the 7.12.1 cell line contained in ATCC Deposit No. PTA-2728;

(i) the antibody expressed by the 7.12.3 cell line contained in ATCC Deposit No. PTA-2728;

(j) the antibody expressed by the 8.3.1 cell line contained in ATCC Deposit No. PTA-2731; and

(k) the antibody expressed by the 8.3.2 cell line contained in ATCC Deposit No. PTA-2731.

57. An antibody that competitively inhibits the binding of the antibody of claim 56 to TR4.

58. A method of inducing apoptosis of cells that express TR4 and TR7 comprising contacting said cells with a first antibody that specifically binds TR4 and a second antibody that specifically binds TR7.

59. The method of claim 58 wherein said first and second antibodies are agonistic antibodies.

60. The method of claim 58 wherein said cells are also contacted with a chemotherapeutic agent.

61. The method of claim 58 wherein said cells are also contacted with TRAIL.

62. The method of claim 58 wherein said cells are in vivo.

63. The method of claim 58 wherein the amount of apoptosis induced by contacting TR4 and TR7 expressing cells with said first and second antibody is greater than the amount of apoptosis induced by contacting TR4 and TR7 expressing cells with either said first or said second antibody alone.

64. The method of claim 58 wherein the antibodies contact the cells simultaneously.

65. The method of claim 58 wherein the antibodies contact the cells sequentially.