PRLR-Specific Antibody and Uses Thereof

• US 20110150760A1
• Filed: 11/22/2010
• Published: 06/23/2011
• Est. Priority Date: 08/18/2006
• Status: Active Grant

• Current Assignee: Novartis Ag, Xoma Technology Ltd. (Xoma Corp.)
• Original Assignee: Novartis Ag, Xoma Technology Ltd. (Xoma Corp.)
• Inventors: MIRZA, AMER, DAMIANO, JASON, BEDINGER, DANIEL, NONET, GENEVIEVE, MASAT, LINDA, LUQMAN, MOHAMMAD
• Granted Patent: US 9,005,614 B2
• Time in Patent Office: Days
• US Class Current: 424/1.49
• 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 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 39/39558 against tumor tissues, cells, antigens
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 47/6851 the antibody targeting a determinant of a tumour cell
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 16/2869 against hormone receptors for antibodies against neuromediator receptors C07K16/286
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/34 Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/73 Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/732 Antibody-dependent cellular cytotoxicity [ADCC]
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 : Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide
• CPC Class Codes: G01N 2333/72 : for hormones for neuromediators G01N2333/70571
• CPC Class Codes: G01N 33/5308 : for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
• CPC Class Codes: G01N 33/57492 : involving compounds localized on the membrane of tumor or cancer cells

Abstract

PRLR-specific antibodies are provided, along with pharmaceutical compositions containing such antibody, kits containing a pharmaceutical composition, and methods of preventing and treating cancer.

Claims

1. An antibody that binds the extracellular domain of PRLR with an equilibrium dissociation constant (K_D) of 10⁻

6 M or lower and competes with any of antibodies chXHA.06.642, chXHA.06.275, he.06.642-1, he.06.642-2, he.06.275-1, he.06.275-2, he.06.275-3, he.06.275-4, XPA.06.128, XPA.06.129, XPA.06.130, XPA.06.131, XPA.06.141, XPA.06.147, XPA.06.148, XPA.06.158, XPA.06.159, XPA.06.163, XPA.06.167, XPA.06.171, XPA.06.178, XPA.06.181, XPA.06.192, XPA.06.202, XPA.06.203, XPA.06.206, XPA.06.207, XPA.06.210, XPA.06.212, XPA.06.217, XPA.06.219, XPA.06.229, XPA.06.233, XPA.06.235, XPA.06.239, XPA.06.145, XHA.06.567, XHA.06.642, XHA.06.983, XHA.06.275, XHA.06.189, or XHA.06.907 for binding to PRLR by more than 75%.

2. The antibody of claim 1 that binds to the same epitope of PRLR as any of antibodies chXHA.06.642, chXHA.06.275, he.06.642-1, he.06.642-2, he.06.275-1, he.06.275-2, he.06.275-3, he.06.275-4, XPA.06.128, XPA.06.129, XPA.06.130, XPA.06.131, XPA.06.141, XPA.06.147, XPA.06.148, XPA.06.158, XPA.06.159, XPA.06.163, XPA.06.167, XPA.06.171, XPA.06.178, XPA.06.181, XPA.06.192, XPA.06.202, XPA.06.203, XPA.06.206, XPA.06.207, XPA.06.210, XPA.06.212, XPA.06.217, XPA.06.219, XPA.06.229, XPA.06.233, XPA.06.235, XPA.06.239, XPA.06.145, XHA.06.567, XHA.06.642, XHA.06.983, XHA.06.275, XHA.06.189, or XHA.06.907.

4. The antibody of any of claims 1-3 wherein the antibody is a chimeric antibody, a humanized antibody, a human engineered antibody, a human antibody, a single chain antibody or an antibody fragment.

5. The antibody of any of the preceding claims in which at least one amino acid within a CDR is substituted by a corresponding residue of a corresponding CDR of another anti-PRLR antibody.

6. The antibody of any of the preceding claims in which one or two amino acids within a CDR have been modified.

7. The antibody of any of the preceding claims that retains at least 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identity over either the variable light or heavy region to the antibodies of chXHA.06.642, chXHA.06.275, he.06.642-1, he.06.642-2, he.06.275-1, he.06.275-2, he.06.275-3, he.06.275-4, XPA.06.128, XPA.06.129, XPA.06.130, XPA.06.131, XPA.06.141, XPA.06.147, XPA.06.148, XPA.06.158, XPA.06.159, XPA.06.163, XPA.06.167, XPA.06.171, XPA.06.178, XPA.06.181, XPA.06.192, XPA.06.202, XPA.06.203, XPA.06.206, XPA.06.207, XPA.06.210, XPA.06.212, XPA.06.217, XPA.06.219, XPA.06.229, XPA.06.233, XPA.06.235, XPA.06.239, XPA.06.145, XHA.06.567, XHA.06.642, XHA.06.983, XHA.06.275, XHA.06.189, or XHA.06.907.

8. The antibody of any of the preceding claims that comprises a constant region of a human antibody sequence and one or more heavy and light chain variable framework regions of a human antibody sequence.

9. The antibody of claim 8 wherein the human antibody sequence is an individual human sequence, a human consensus sequence, an individual human germline sequence, or a human consensus germline sequence.

10. The antibody of any of the preceding claims wherein the heavy chain constant region is a modified or unmodified IgG, IgM, IgA, IgD, IgE, a fragment thereof, or combinations thereof.

11. The antibody of claim 10 wherein the heavy chain constant region is a modified or unmodified IgG1, IgG2, IgG3 or IgG4.

12. The antibody of any of the preceding claims that has an equilibrium dissociation constant of 10⁻

6, 10<sup>−

7</sup>, 10<sup>−

8</sup> or 10<sup>−

9</sup> M or lower to PRLR.

13. The antibody of any of the preceding claims comprising a conservative substitution in the CDRs.

14. The antibody of any of the preceding claims comprising a conservative or non-conservative change in low and moderate risk residues.

15. The antibody of any of the preceding claims wherein the light chain constant region is a modified or unmodified lambda light chain constant region , a kappa light chain constant region, a fragment thereof, or combinations thereof.

16. The antibody of any of the preceding claims that inhibits PRLR dimerization.

17. The antibody of any of the preceding claims that inhibits PRLR intracellular phosphorylation.

18. The antibody of any of the preceding claims that inhibits the induction of MAPK phosphorylation.

19. The antibody of any of the preceding claims that inhibits the induction of Stat5 phosphorylation.

20. The antibody of any of the preceding claims that inhibits the induction of AKT phosphorylation.

21. The antibody of any of the preceding claims that inhibits the binding of PRL to PRLR.

22. The antibody of any of the preceding claims that inhibits VEGF production and/or angiogenesis.

23. The antibody of any of the preceding claims that inhibits the proliferation of a cancer cell.

24. The antibody of claim 23 that inhibits proliferation of a breast, prostate, or lung cancer cell.

25. The antibody of any of the preceding claims that is conjugated to another diagnostic or therapeutic agent.

29. A method of treating a subject suffering from cancer comprising the step of administering an antibody of any of the preceding claims in a therapeutically effective amount.

30. The method of claim 29 wherein the cancer is breast, lung or prostate cancer.

31. The method of claim 30 wherein a second therapeutic agent is administered.

32. The method of claim 30 wherein the second therapeutic agent is doxorubicin or daunorubicin.

33. The method of claim 32 wherein the antibody is chXHA.06.642, chXHA.06.275, he.06.642-1, he.06.642-2, he.06.275-1, he.06.275-2, he.06.275-3, he.06.275-4.

34. The method of claim 30 wherein the subject is positive for PRLR expression and HER2-neu expression, and wherein said second therapeutic agent is an anti-Her2-neu antibody.

35. The method of claim 30 wherein the subject is positive for PRLR expression and ER expression, and wherein said second therapeutic agent is an anti-ER antibody.

36. The method of claim 30 wherein the subject is further treated with radiation therapy or surgery.

37. A method of targeting a tumor cell expressing PRLR comprising the step of administering an antibody of any of the preceding claims, conjugated to a radionuclide or other toxin.

38. The method of claims 30-37 wherein the subject is a mammal.

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

40. An isolated nucleic acid molecule comprising a nucleotide sequence that encodes the heavy chain or light chain of an antibody of any of claims 1-23.

41. An expression vector comprising the nucleic acid molecule of claim 40 operably linked to a regulatory control sequence.

42. A host cell comprising the vector of claim 41 or a nucleic acid molecule of claim 40.

43. A method of using the host cell of claim 42 to produce an antibody, comprising culturing the host cell of claim 36 under suitable conditions and recovering said antibody.

44. An antibody produced by the method of claim 43.

45. The antibody of any of claim 1-23 or 44 that is purified to at least 95% homogeneity by weight.

46. A pharmaceutical composition comprising the antibody of claim 45 and a pharmaceutically acceptable carrier.

47. A kit comprising an antibody of any of the preceding claims comprising a therapeutically effective amount of an antibody of the invention, packaged in a container, said kit optionally containing a second therapeutic agent, and further comprising a label attached to or packaged with the container, the label describing the contents of the container and providing indications and/or instructions regarding use of the contents of the container to treat cancer.

48. The kit of claim 47 wherein the container is a vial or bottle or prefilled syringe.

66. A method of identifying a subject in need of treatment with an anti-PRLR antibody according to any one of claims 1-23 and 49-65 comprising the steps of:

(a) obtaining a sample from the subject; and

(b) analyzing the sample for level of phosphorylation of PRLR, Jak2, Mapk, Stat5, Erk1/2 and/or Akt;

wherein the level of phosphorylation of PRLR, Jak2, Mapk, Stat5, Erk1/2 and/or Akt is indicative of a need for treatment with an anti-PRLR antibody.

68. The method according to claim 67 wherein the cancer therapeutic is an antibody according to any one of claims 1-23 and 49-65.

3. An antibody that comprises 1, 2, 3, 4, 5 or 6 CDRs of any of antibodies chXHA.06.642, chXHA.06.275, he.06.642-1, he.06.642-2, he.06.275-1, he.06.275-2, he.06.275-3, he.06.275-4, XPA.06.128, XPA.06.129, XPA.06.130, XPA.06.131, XPA.06.141, XPA.06.147, XPA.06.148, XPA.06.158, XPA.06.159, XPA.06.163, XPA.06.167, XPA.06.171, XPA.06.178, XPA.06.181, XPA.06.192, XPA.06.202, XPA.06.203, XPA.06.206, XPA.06.207, XPA.06.210, XPA.06.212, XPA.06.217, XPA.06.219, XPA.06.229, XPA.06.233, XPA.06.235, XPA.06.239, XPA.06.145, XHA.06.567, XHA.06.642, XHA.06.983, XHA.06.275, XHA.06.189, or XHA.06.907.

26. A method of screening for an antibody to the extracellular domain of a PRLR protein useful for the treatment of cancer comprising the steps of:

contacting a polypeptide comprising the ECD of PRLR with a candidate antibody that contains at least 1, 2, 3, 4, 5, or 6 CDRs of antibodies chXHA.06.642, chXHA.06.275, he.06.642-1, he.06.642-2, he.06.275-1, he.06.275-2, he.06.275-3, he.06.275-4, XPA.06.128, XPA.06.129, XPA.06.130, XPA.06.131, XPA.06.141, XPA.06.147, XPA.06.148, XPA.06.158, XPA.06.159, XPA.06.163, XPA.06.167, XPA.06.171, XPA.06.178, XPA.06.181, XPA.06.192, XPA.06.202, XPA.06.203, XPA.06.206, XPA.06.207, XPA.06.210, XPA.06.212, XPA.06.217, XPA.06.219, XPA.06.229, XPA.06.233, XPA.06.235, XPA.06.239, XPA.06.145, XHA.06.567, XHA.06.642, XHA.06.983, XHA.06.275, XHA.06.189, and XHA.06.907;

detecting binding affinity of the candidate antibody to the polypeptide, and

identifying said candidate antibody as an antibody useful for the treatment of cancer if an equilibrium dissociation constant of 10<sup>−

6</sup> M or lower is detected.

27. A method of systematically altering antibodies and screening for an antibody to the extracellular domain of a PRLR protein useful for the treatment of cancer comprising the steps of:

preparing a candidate antibody that contains modifications to one or two amino acids within the CDRs of antibodies chXHA.06.642, chXHA.06.275, he.06.642-1, he.06.642-2, he.06.275-1, he.06.275-2, he.06.275-3, he.06.275-4, XPA.06.128, XPA.06.129, XPA.06.130, XPA.06.131, XPA.06.141, XPA.06.147, XPA.06.148, XPA.06.158, XPA.06.159, XPA.06.163, XPA.06.167, XPA.06.171, XPA.06.178, XPA.06.181, XPA.06.192, XPA.06.202, XPA.06.203, XPA.06.206, XPA.06.207, XPA.06.210, XPA.06.212, XPA.06.217, XPA.06.219, XPA.06.229, XPA.06.233, XPA.06.235, XPA.06.239, XPA.06.145, XHA.06.567, XHA.06.642, XHA.06.983, XHA.06.275, XHA.06.189, and XHA.06.907,

contacting a polypeptide comprising the ECD of PRLR with said candidate antibody

detecting binding affinity of the candidate antibody to the polypeptide, and

identifying said candidate antibody as an antibody useful for the treatment of cancer if an equilibrium dissociation constant of 10<sup>−

6</sup> M or lower is detected.

28. A method of screening for an antibody to the extracellular domain of a PRLR protein useful for the treatment of cancer comprising the steps of:

contacting a breast, lung, or prostate cell with a candidate antibody that contains at least 1, 2, 3, 4, 5 or 6 CDRs of antibodies chXHA.06.642, chXHA.06.275, he.06.642-1, he.06.642-2, he.06.275-1, he.06.275-2, he.06.275-3, he.06.275-4, XPA.06.128, XPA.06.129, XPA.06.130, XPA.06.131, XPA.06.141, XPA.06.147, XPA.06.148, XPA.06.158, XPA.06.159, XPA.06.163, XPA.06.167, XPA.06.171, XPA.06.178, XPA.06.181, XPA.06.192, XPA.06.202, XPA.06.203, XPA.06.206, XPA.06.207, XPA.06.210, XPA.06.212, XPA.06.217, XPA.06.219, XPA.06.229, XPA.06.233, XPA.06.235, XPA.06.239, XPA.06.145, XHA.06.567, XHA.06.642, XHA.06.983, XHA.06.275, XHA.06.189, and XHA.06.907 or an antibody that contains a modification of one or two amino acids within one or more CDRs;

detecting proliferation or survival of said cell; and

identifying said candidate antibody as an antibody useful for the treatment of cancer if a decrease in cell proliferation or survival is detected.

49. An antibody that binds the extracellular domain of PRLR comprising a variable light chain amino acid sequence selected from the group consisting of SEQ ID NO:

21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 82, 84, 86, 88, 91, 95 and 96.

50. An antibody that binds the extracellular domain of PRLR with comprising a variable heavy chain amino acid sequence selected from the group consisting of SEQ ID NO:

20, 2, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 83, 85, 87, 89,90, 93, 94, 97 and 98.

51. An antibody that binds the extracellular domain of PRLR comprising a variable light chain amino acid sequence SEQ ID NO:

88, and a variable heavy chain amino acid sequence of SEQ ID NO;

89.

52. An antibody that binds the extracellular domain of PRLR comprising a variable light chain amino acid sequence SEQ ID NO:

88, and a variable heavy chain amino acid sequence of SEQ ID NO;

90.

53. An antibody that binds the extracellular domain of PRLR comprising a variable light chain amino acid sequence of SEQ ID NO:

91, and a variable heavy chain amino acid sequence of SEQ ID NO;

93.

54. An antibody that binds the extracellular domain of PRLR comprising a variable light chain amino acid sequence of SEQ ID NO:

91, and a variable heavy chain amino acid sequence of SEQ ID NO;

94.

55. An antibody that binds the extracellular domain of PRLR comprising a variable light chain amino acid sequence of SEQ ID NO:

92, and a variable heavy chain amino acid sequence of SEQ ID NO;

93.

56. An antibody that binds the extracellular domain of PRLR comprising a variable light chain amino acid sequence of SEQ ID NO:

92, and a variable heavy chain amino acid sequence of SEQ ID NO;

94.

57. An antibody that binds the extracellular domain of PRLR comprising a variable light chain amino acid sequence of SEQ ID NO:

95, and a variable heavy chain amino acid sequence of SEQ ID NO;

97.

58. An antibody that binds the extracellular domain of PRLR comprising a variable light chain amino acid sequence of SEQ ID NO:

95, and a variable heavy chain amino acid sequence of SEQ ID NO;

98.

59. An antibody that binds the extracellular domain of PRLR comprising a variable light chain amino acid sequence of SEQ ID NO:

96, and a variable heavy chain amino acid sequence of SEQ ID NO;

97.

60. An antibody that binds the extracellular domain of PRLR comprising a variable light chain amino acid sequence of SEQ ID NO:

96, and a variable heavy chain amino acid sequence of SEQ ID NO;

98.

61. An antibody that binds the extracellular domain of human PRLR with an equilibrium dissociation constant (K_D) of at least 10,000 to 15,000 fold lower than the extracellular domain of murine PRLR.

62. The antibody of claim 61 that binds the same epitope as he.06.275-4.

63. An antibody that binds the extracellular domain of human PRLR, the extracellular domain of murine PRLR, and the extracellular domain of rat PRLR

64. An antibody that binds the extracellular domain of human, murine and rat PRLR with an equilibrium dissociation constant (K_D) of 10⁻

6 M or lower.

65. The antibody of claim 64 that binds the same epitope as he.06.642-2.

67. A method of monitoring cancer therapy in a subject afflicted with cancer comprising the steps of:

(a) analyzing a first sample from the subject for level of phosphorylation of PRLR prior to the initiation of treatment with a cancer therapeutic; and

(b) analyzing a second sample after the initiation of the treatment with the cancer therapeutic,

wherein a reduction in the level of phosphorylated PRLR after the initiation of the treatment with the cancer therapeutic indicates the patient is receiving a therapeutically effective dose of the cancer therapeutic.