Human Monoclonal Antibodies To Programmed Death 1(PD-1) And Methods For Treating Cancer Using Anti-PD-1 Antibodies Alone or in Combination with Other Immunotherapeutics

• US 20090217401A1
• Filed: 05/02/2006
• Published: 08/27/2009
• Est. Priority Date: 05/09/2005
• Status: Active Grant

• Current Assignee: ER Squibb & Sons LLC (Bristol-Myers Squibb Company)
• Original Assignee: Medarex Incorporated (Bristol-Myers Squibb Company)
• Inventors: Chen, Bing, Cardarelli, Josephine M., Srinivasan, Mohan, Korman, Alan J., Wang, Changyu, Selby, Mark J.
• Granted Patent: US 8,008,449 B2
• Time in Patent Office: Days
• US Class Current: 800/18
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2039/507 Comprising a combination of two or more separate antibodies
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• 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 47/6849 the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
• CPC Class Codes: A61K 51/10 Antibodies or immunoglobulins; Fragments thereof , the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 16/2803 against the immunoglobulin superfamily
• CPC Class Codes: C07K 16/2818 against CD28 or CD152
• CPC Class Codes: C07K 16/468 Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
• 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 2317/33 Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
• 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 2317/73 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 2317/74 : Inducing cell proliferation
• CPC Class Codes: C07K 2317/75 : Agonist effect on antigen
• 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 2317/94 : Stability, e.g. half-life, pH, temperature or enzyme-resistance
• CPC Class Codes: Y02A 50/30 : Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The present invention provides isolated monoclonal antibodies, particularly human monoclonal antibodies, that specifically bind to PD-1 with high affinity. Nucleic acid molecules encoding the antibodies of the invention, expression vectors, host cells and methods for expressing the antibodies of the invention are also provided. Immunoconjugates, bispecific molecules and pharmaceutical compositions comprising the antibodies of the invention are also provided. The invention also provides methods for detecting PD-1, as well as methods for treating various diseases, including cancer and infectious diseases, using anti-PD-1 antibodies. The present invention further provides methods for using a combination immunotherapy, such as the combination of anti-CTLA-4 and anti-PD-1 antibodies, to treat hyperproliferative disease, such as cancer. The invention also provides methods for altering adverse events related to treatment with such antibodies individually.

Claims

1. An isolated human monoclonal antibody, or an antigen-binding portion thereof;

wherein the antibody binds to PD-1 and wherein the antibody exhibits at least one of the following properties;

a) binds to human PD-1 with a K_D of 1×

10<sup>−

7</sup>M or less;

b) does not substantially bind to human CD28, CTLA-4 or ICOS;

c) increases T-cell proliferation in a Mixed Lymphocyte Reaction (MLR) assay;

d) increases interferon-gamma production in an MLR assay;

or

e) increases interleukin-2 (IL-2) secretion in an MLR assay.

43. A composition comprising the antibody, or antigen-binding portion thereof, of claim 1, and a pharmaceutically acceptable carrier.

44. An immunoconjugate comprising the antibody, or antigen-binding portion thereof, of claim 1, linked to a therapeutic agent.

50. A bispecific molecule comprising the antibody, or antigen-binding portion thereof, of claim 1, linked to a second functional moiety having a different binding specificity than said antibody, or antigen-binding portion thereof.

52. An isolated nucleic acid molecule encoding the antibody, or antigen-binding portion thereof, of claim 1.

53. An expression vector comprising the nucleic acid molecule of claim 52.

54. A host cell comprising the expression vector of claim 53.

55. A transgenic mouse comprising human immunoglobulin heavy and light chain transgenes, wherein the mouse expresses the antibody of claim 1.

56. A hybridoma prepared from the mouse of claim 55, wherein the hybridoma produces said antibody.

57. A method of modulating an immune response in a subject comprising administering to the subject the antibody, or antigen-binding portion thereof, of claim 1 such that the immune response in the subject is modulated.

62. The method of claim 64, wherein the tumor cells are of a cancer selected from the group consisting of melanoma, renal cancer, prostate cancer, breast cancer, colon cancer and lung cancer.

64. A method of inhibiting growth of tumor cells in a subject, comprising administering to a subject the antibody, or antigen-binding portion thereof, of claim 1 in an amount effective to inhibit growth of the tumor cells.

65. A method of treating an infectious disease in a subject comprising administering to the subject the antibody, or antigen-binding portion thereof, of claim 1 such that the subject is treated for the infectious disease.

67. A method of enhancing an immune response to an antigen in a subject, comprising administering to the subject:

(i) the antigen; and

(ii) the antibody, or antigen-binding portion thereof, of claim 1 such that an immune response to the antigen in the subject is enhanced.

68. The method of claim 67, wherein the antigen is a tumor antigen, a viral antigen, a bacterial antigen or an antigen from a pathogen.

2-7. -7. (canceled)

8. An isolated human monoclonal antibody, or an antigen-binding portion thereof;

wherein the antibody cross-competes for binding to PD-1 with a reference antibody, wherein the reference antibody is selected from the group consisting of;

a) an antibody comprising a human heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

1 and a human light chain variable region comprising the amino acid sequence of SEQ ID NO;

8;

b) an antibody comprising a human heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

2 and a human light chain variable region comprising the amino acid sequence of SEQ ID NO;

9;

c) an antibody comprising a human heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

3 and a human light chain variable region comprising the amino acid sequence of SEQ ID NO;

10;

d) an antibody comprising a human heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

4 and a human light chain variable region comprising the amino acid sequence of SEQ ID NO;

11;

e) an antibody comprising a human heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

5 and the human light chain variable region comprising the amino acid sequence of SEQ ID NO;

12;

f) an antibody comprising a human heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

6 and the human light chain variable region comprising the amino acid sequence of SEQ ID NO;

13; and

g) an antibody comprising a human heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

7 and a human light chain variable region comprising the amino acid sequence of SEQ ID NO;

14.

9-26. -26. (canceled)

27. An isolated monoclonal antibody, or antigen-binding portion thereof, wherein the antibody is selected from the group consisting of:

a) an antibody comprising

a heavy chain variable region CDR1 comprising SEQ ID NO;

15;

a heavy chain variable region CDR2 comprising SEQ ID NO;

22;

a heavy chain variable region CDR3 comprising SEQ ID NO;

29;

a light chain variable region CDR1 comprising SEQ ID NO;

36;

a light chain variable region CDR2 comprising SEQ ID NO;

43; and

a light chain variable region CDR3 comprising SEQ ID NO;

50;

b) an antibody comprising

a heavy chain variable region CDR1 comprising SEQ ID NO;

16;

a heavy chain variable region CDR2 comprising SEQ ID NO;

23;

a heavy chain variable region CDR3 comprising SEQ ID NO;

30;

a light chain variable region CDR1 comprising SEQ ID NO;

37;

a light chain variable region CDR2 comprising SEQ ID NO;

44; and

a light chain variable region CDR3 comprising SEQ ID NO;

51;

c) an antibody comprising

a heavy chain variable region CDR1 comprising SEQ ID NO;

17;

a heavy chain variable region CDR2 comprising SEQ ID NO;

24;

a heavy chain variable region CDR3 comprising SEQ ID NO;

31;

a light chain variable region CDR1 comprising SEQ ID NO;

38;

a light chain variable region CDR2 comprising SEQ ID NO;

45; and

a light chain variable region CDR3 comprising SEQ ID NO;

52;

d) an antibody comprising

a heavy chain variable region CDR1 comprising SEQ ID NO;

18;

a heavy chain variable region CDR2 comprising SEQ ID NO;

25;

a heavy chain variable region CDR3 comprising SEQ ID NO;

32;

a light chain variable region CDR1 comprising SEQ ID NO;

39;

a light chain variable region CDR2 comprising SEQ ID NO;

46; and

a light chain variable region CDR3 comprising SEQ ID NO;

53

e) an antibody comprising

a heavy chain variable region CDR1 comprising SEQ ID NO;

19;

a heavy chain variable region CDR2 comprising SEQ ID NO;

26;

a heavy chain variable region CDR3 comprising SEQ ID NO;

33;

a light chain variable region CDR1 comprising SEQ ID NO;

40;

a light chain variable region CDR2 comprising SEQ ID NO;

47; and

a light chain variable region CDR3 comprising SEQ ID NO;

54

f) an antibody comprising

a heavy chain variable region CDR1 comprising SEQ ID NO;

20;

a heavy chain variable region CDR2 comprising SEQ ID NO;

27;

a heavy chain variable region CDR3 comprising SEQ ID NO;

34;

a light chain variable region CDR1 comprising SEQ ID NO;

41;

a light chain variable region CDR2 comprising SEQ ID NO;

48; and

a light chain variable region CDR3 comprising SEQ ID NO;

55; and

g) an antibody comprising

a heavy chain variable region CDR1 comprising SEQ ID NO;

21;

a heavy chain variable region CDR2 comprising SEQ ID NO;

28;

a heavy chain variable region CDR3 comprising SEQ ID NO;

35;

a light chain variable region CDR1 comprising SEQ ID NO;

42;

a light chain variable region CDR2 comprising SEQ ID NO;

49; and

a light chain variable region CDR3 comprising SEQ ID NO;

56,

wherein the antibody specifically binds PD-1.

28-34. -34. (canceled)

35. An isolated monoclonal antibody, or antigen-binding portion thereof, wherein the antibody is selected from the group consisting of:

a) an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

1; and

a light chain variable region comprising the amino acid sequence of SEQ ID NO;

8;

b) an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

2; and

a light chain variable region comprising the amino acid sequence of SEQ ID NO;

9;

c) an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

3; and

a light chain variable region comprising the amino acid sequence of SEQ ID NO;

10;

d) an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

4; and

a light chain variable region comprising the amino acid sequence of SEQ ID NO;

11;

e) an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

5; and

a light chain variable region comprising the amino acid sequence of SEQ ID NO;

12;

or

f) an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

6; and

a light chain variable region comprising the amino acid sequence of SEQ ID NO;

13; and

g) an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

7; and

a light chain variable region comprising the amino acid sequence of SEQ ID NO;

14,

wherein the antibody specifically binds PD-1.

36-42. -42. (canceled)

45-49. -49. (canceled)

51. (canceled)

58-61. -61. (canceled)

63. (canceled)

66. (canceled)

69. A method for treating a hyperproliferative disease, which method comprises:

(a) administering a anti-PD-1 antibody to a subject, and

(b) administering a anti-CTLA-4 antibody to the subject.

70. The method of claim 69, wherein the anti-PD-1 antibody is administered at a subtherapeutic dose.

71. The method of claim 69, wherein the anti-CTLA-4 antibody is administered at a subtherapeutic dose.

72. The method of claim 69, wherein the anti-PD-1 antibody and the anti-CTLA-4 antibody are each administered at a subtherapeutic dose.

79. The method of claim 69 wherein the hyperproliferative disease is cancer.

80. The method of claim 79 wherein the cancer is colon cancer.

84. The method of claim 69, wherein the anti-PD-1 human sequence monoclonal antibody is selected from the group consisting of:

a) an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

1; and

a light chain variable region comprising the amino acid sequence of SEQ ID NO;

8;

b) an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

2; and

a light chain variable region comprising the amino acid sequence of SEQ ID NO;

9;

c) an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

3; and

a light chain variable region comprising the amino acid sequence of SEQ ID NO;

10;

d) an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

4; and

a light chain variable region comprising the amino acid sequence of SEQ ID NO;

11;

e) an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

5; and

a light chain variable region comprising the amino acid sequence of SEQ ID NO;

12;

or

f) an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

6; and

a light chain variable region comprising the amino acid sequence of SEQ ID NO;

13; and

g) an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

7; and

a light chain variable region comprising the amino acid sequence of SEQ ID NO;

14.

73-78. -78. (canceled)

81-83. -83. (canceled)

85-87. -87. (canceled)

88. A method for altering an adverse event associated with treatment of a hyperproliferative disease with an immunostimulatory agent, which method comprises:

(a) administering an anti-PD-1 antibody to a subject, and

(b) administering a subtherapeutic dose of anti-CTLA-4 antibody to the subject.

96. The method of claim 88, wherein the hyperproliferative disease is cancer.

97. The method of claim 96 wherein the cancer is colon cancer.

101. The method of claim 88 wherein the anti-PD-1 human sequence monoclonal antibody is selected from the group consisting of:

a) an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

1; and

a light chain variable region comprising the amino acid sequence of SEQ ID NO;

8;

b) an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

2; and

a light chain variable region comprising the amino acid sequence of SEQ ID NO;

9;

c) an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

3; and

a light chain variable region comprising the amino acid sequence of SEQ ID NO;

10;

d) an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

4; and

a light chain variable region comprising the amino acid sequence of SEQ ID NO;

11;

e) an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

5; and

a light chain variable region comprising the amino acid sequence of SEQ ID NO;

12;

or

f) an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

6; and

a light chain variable region comprising the amino acid sequence of SEQ ID NO;

13; and

g) an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO;

7; and

a light chain variable region comprising the amino acid sequence of SEQ ID NO;

14.

90-95. -95. (canceled)

98-100. -100. (canceled)

102-104. -104. (canceled)

105. A method for preparing an anti-PD-1 antibody comprising:

(a) providing;

(i) a heavy chain variable region antibody sequence comprising a CDR1 sequence that is selected from the group consisting of SEQ ID NOs;

15, 16, 17, 18, 19, 20 and 21, a CDR2 sequence that is selected from the group consisting of SEQ ID NOs;

22, 23, 24, 25, 26, 27 and 28; and

a CDR3 sequence that is selected from the group consisting of SEQ ID NOs;

29, 30, 31, 32, 33, 34 and 35;

or (ii) a light chain variable region antibody sequence comprising a CDR1 sequence that is selected from the group consisting of SEQ ID NOs;

36, 37, 38, 39, 40, 41 and 42, a CDR2 sequence that is selected from the group consisting of SEQ ID NOs;

43, 44, 45, 46, 47, 48 and 49, and a CDR3 sequence that is selected from the group consisting of SEQ ID NOs;

50, 51, 52, 53, 54, 55 and 56;

(b) altering at least one amino acid residue within at least one variable region antibody sequence, said sequence being selected from the heavy chain variable region antibody sequence and the light chain variable region antibody sequence, to create at least one altered antibody sequence; and

(c) expressing the altered antibody sequence as a protein.