METHODS FOR TREATING CANCER USING ANTI-PD-1 ANTIBODIES IN COMBINATION WITH ANTI-CTLA-4 ANTIBODIES

• US 20140212422A1
• Filed: 03/27/2014
• Published: 07/31/2014
• Est. Priority Date: 05/09/2005
• Status: Active Grant

• Current Assignee: ER Squibb & Sons LLC (Bristol-Myers Squibb Company), ONO Pharmaceutical Company Limited
• Original Assignee: Medarex LLC (Bristol-Myers Squibb Company), ONO Pharmaceutical Company Limited
• Inventors: Korman, Alan J., Srinivasan, Mohan, Wang, Changyu, Selby, Mark J., Chen, Bingliang, Cardarelli, Josephine M., Huang, Haichun
• Granted Patent: US 9,358,289 B2
• Time in Patent Office: Days
• US Class Current: 424/135.1
• 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. A method for treating a cancer in a subject comprising administering to the subject a combination of an anti-PD-1 antibody, or an antigen-binding portion thereof, and an anti-CTLA-4 antibody, or an antigen-binding portion thereof, wherein the combination of the anti-PD-1 antibody or antigen-binding portion thereof and the anti-CTLA-4 antibody or antigen-binding portion thereof exhibits a synergistic therapeutic effect on cancer treatment.

2. The method of claim 1, wherein the anti-PD-1 antibody or antigen-binding portion thereof is a monoclonal antibody or an antigen-binding portion thereof that binds specifically to PD-1.

3. The method of claim 2, wherein the subject is a human and the anti-PD-1 monoclonal antibody or antigen-binding portion thereof binds specifically to human PD-1.

14. The method of any one of claims 1, 4, 8 and 10, wherein the anti-PD-1 antibody or antigen-binding portion thereof is a chimeric monoclonal antibody or an antigen-binding portion thereof.

16. The method of any one of claims 1, 4, 8 and 10, wherein the anti-PD-1 antibody or antigen-binding portion thereof is a humanized monoclonal antibody or an antigen-binding portion thereof.

18. The method of any one of claims 1, 4, 8 and 10, wherein the anti-PD-1 antibody or antigen-binding portion thereof is a human monoclonal antibody or an antigen-binding portion thereof.

20. The method of any one of claims 1, 4, 8 and 10, wherein the anti-PD-1 antibody or antigen-binding portion thereof comprises a heavy chain constant region which is of a human IgG 1 or IgG4 isotype.

22. The method of any one of claims 1, 4, 8 and 10, wherein the anti-PD-1 antibody or antigen-binding portion thereof comprises a light chain constant region which is a human kappa or lambda constant region.

24. The method of any one of claims 1, 4, 8 and 10, wherein the anti-PD-1 antigen-binding portion is a Fab, Fab′

, F(ab′

)₂, dAb, Fd, Fv, a single chain Fv fragment or an isolated CDR.

26. The method of claim 1, wherein the anti-CTLA-4 antibody or antigen-binding portion thereof is a monoclonal antibody that binds specifically to CTLA-4.

27. The method of claim 26, wherein the subject is a human and the anti-CTLA-4 monoclonal antibody or antigen-binding portion thereof binds specifically to human CTLA-4.

28. The method of claim 27, wherein the anti-CTLA-4 antibody or antigen-binding portion thereof is a human monoclonal antibody or an antigen-binding portion thereof.

29. The method of any one of claims 1, 4, 8 and 10, wherein the anti-CTLA-4 antibody is the antibody designated 10D 1.

30. The method of any one of claims 1, 4, 8 and 10, wherein the cancer is selected from the group consisting of melanoma, lung cancer, renal cancer, prostate cancer, breast cancer, colorectal cancer and fibrosarcoma.

31. The method of any one of claims 1, 4, 8 and 10, wherein the cancer is selected from the group consisting of bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin'"'"'s Disease, non-Hodgkin'"'"'s lymphoma, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, chronic or acute leukemias including acute myeloid leukaemia, chronic myeloid leukaemia, acute lymphoblastic leukaemia, chronic lymphocytic leukaemia, solid tumors of childhood, lymphocytic lymphoma, cancer of the bladder, cancer of the kidney or ureter, carcinoma of the renal pelvis, neoplasm of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, Kaposi'"'"'s sarcoma, epidermoid cancer, squamous cell cancer, T-cell lymphoma, environmentally induced cancers including those induced by asbestos, and combinations of said cancers.

38. The method of any one of claims 1, 4, 8, 10 and 32, wherein the anti-PD-1 antibody or antigen-binding portion thereof and the anti-CTLA-4 antibody or antigen-binding portion thereof are administered sequentially to the subject.

39. The method of claim 38, wherein

(a) the anti-PD-1 antibody or antigen-binding portion thereof is administered before the anti-CTLA-4 antibody or antigen-binding portion thereof;

or

(b) the anti-CTLA-4 antibody or antigen-binding portion thereof is administered before the anti-PD-1 antibody or antigen-binding portion thereof.

40. The method of any one of claims 1, 4, 8, 10 and 32, wherein the anti-PD-1 antibody or antigen-binding portion thereof and the anti-CTLA-4 antibody or antigen-binding portion thereof are administered concurrently in separate compositions.

41. The method of any one of claims 1, 4, 8, 10 and 32, wherein the anti-PD-1 antibody or antigen-binding portion thereof and the anti-CTLA-4 antibody or antigen-binding portion thereof are admixed as a single composition and administered concurrently.

42. The method of any one of claims 1, 4, 8, 10 and 32, wherein the anti-PD-1 antibody or antigen-binding portion thereof is administered at a subtherapeutic dose.

43. The method of any one of claims 1, 4, 8, 10 and 32, wherein the anti-CTLA-4 antibody or antigen-binding portion thereof is administered at a subtherapeutic dose.

44. The method of any one of claims 1, 4, 8, 10 and 32, wherein the anti-PD-1 antibody or antigen-binding portion thereof and the anti-CTLA-4 antibody or antigen-binding portion thereof are each administered at a subtherapeutic dose.

45. The method of any one of claims 1, 4, 8, 10 and 32, wherein the anti-PD-1 antibody is administered to the subject at a dosage range of from about 1 mg/kg to about 10 mg/kg.

46. The method of any one of claims 1, 4, 8, 10 and 32, wherein the anti-CTLA-4 antibody is administered to the subject at a dosage range of from about 0.2 mg/kg to about 10 mg/kg.

4. A method for treating a cancer in a subject comprising administering to the subject a combination of an anti-PD-1 monoclonal antibody, or an antigen-binding portion thereof, and an anti-CTLA-4 monoclonal antibody, or an antigen-binding portion thereof, wherein the anti-PD-1 monoclonal antibody or antigen-binding portion thereof cross-competes for binding to PD-1 with a reference antibody, or antigen-binding portion thereof, comprising:

(a) a heavy chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

15;

a heavy chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

22;

a heavy chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

29;

a light chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

36;

a light chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

43; and

a light chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

50;

(b) a heavy chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

16;

a heavy chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

23;

a heavy chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

30;

a light chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

37;

a light chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

44; and

a light chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

51;

(c) a heavy chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

17;

a heavy chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

24;

a heavy chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

31;

a light chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

38;

a light chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

45; and

a light chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

52;

(d) a heavy chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

18;

a heavy chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

25;

a heavy chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

32;

a light chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

39;

a light chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

46; and

a light chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

53;

(e) a heavy chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

19;

a heavy chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

26;

a heavy chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

33;

a light chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

40;

a light chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

47; and

a light chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

54;

(f) a heavy chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

20;

a heavy chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

27;

a heavy chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

34;

a light chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

41;

a light chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

48; and

a light chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

55;

or

(g) a heavy chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

21;

a heavy chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

28;

a heavy chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

35;

a light chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

42;

a light chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

49; and

a light chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

56,

wherein the anti-PD-1 monoclonal antibody or antigen-binding portion thereof exhibits at least one of the following properties;

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

10<sup>−

7</sup> M or less, wherein the K_D is measured by surface plasmon resonance (Biacore) analysis;

(ii) binds to human PD-1 with an on rate (k_on) of about 0.76×

10⁵ l/Ms or more, wherein the on rate (k_on) is measured by surface plasmon resonance (Biacore) analysis; and

(iii) binds to human PD-1 with an off rate (k_off) of about 4.5×

10<sup>−

4</sup> l/s or less, wherein the off rate (k_off) is measured by surface plasmon resonance (Biacore) analysis.

5. The method of claim 4, wherein the reference antibody or antigen-binding portion thereof comprises a heavy chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO:

18;

a heavy chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

25;

a heavy chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

32;

a light chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

39;

a light chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

46; and

a light chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

53.

6. The method of claim 4, wherein the reference antibody or antigen-binding portion thereof comprises:

(a) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

1 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

8;

(b) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

2 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

9;

(c) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

3 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

10;

(d) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

4 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

11;

(e) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

5 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

12;

(f) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

6 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

13;

or

(g) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

7 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

14.

7. The method of claim 6, wherein the reference antibody or antigen-binding portion thereof comprises a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO:

4 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

11.

15. The method of claim 7 or claim 9, wherein the anti-PD-1 antibody or antigen-binding portion thereof is a chimeric monoclonal antibody or an antigen-binding portion thereof.

17. The method of claim 7 or claim 9, wherein the anti-PD-1 antibody or antigen-binding portion thereof is a humanized monoclonal antibody or an antigen-binding portion thereof.

19. The method of claim 7 or claim 9, wherein the anti-PD-1 antibody or antigen-binding portion thereof is a human monoclonal antibody or an antigen-binding portion thereof.

21. The method of claim 7 or claim 9, wherein the anti-PD-1 antibody or antigen-binding portion thereof comprises a heavy chain constant region which is of a human IgG1 or IgG4 isotype.

23. The method of claim 7 or claim 9, wherein the anti-PD-1 antibody or antigen-binding portion thereof comprises a light chain constant region which is a human kappa or lambda constant region.

25. The method of claim 7 or claim 9, wherein the anti-PD-1 antigen-binding portion is a Fab, Fab′

, F(ab′

)₂, dAb, Fd, Fv, a single chain Fv fragment or an isolated CDR.

8. A method for treating a cancer in a subject comprising administering to the subject a combination of an anti-PD-1 monoclonal antibody, or an antigen-binding portion thereof, and an anti-CTLA-4 monoclonal antibody, or an antigen-binding portion thereof, wherein the anti-PD-1 monoclonal antibody or antigen-binding portion thereof binds to the same epitope on human PD-1 as does a reference antibody, or antigen-binding portion thereof, comprising:

(a) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

1 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

8;

(b) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

2 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

9;

(c) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

3 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

10;

(d) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

4 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

11;

(e) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

5 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

12;

(f) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

6 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

13;

or

(g) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

7 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

14.

9. The method of claim 8, wherein the reference antibody or antigen-binding portion thereof comprises a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO:

4 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

11.

10. A method for treating a cancer in a subject comprising administering to the subject a combination of an anti-PD-1 monoclonal antibody, or an antigen-binding portion thereof, and an anti-CTLA-4 monoclonal antibody, or an antigen-binding portion thereof, wherein the anti-PD-1 monoclonal antibody or antigen-binding portion thereof comprises:

(a) a heavy chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

15;

a heavy chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

22;

a heavy chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

29;

a light chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

36;

a light chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

43; and

a light chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

50;

(b) a heavy chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

16;

a heavy chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

23;

a heavy chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

30;

a light chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

37;

a light chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

44; and

a light chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

51;

(c) a heavy chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

17;

a heavy chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

24;

a heavy chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

31;

a light chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

38;

a light chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

45; and

a light chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

52;

(d) a heavy chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

18;

a heavy chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

25;

a heavy chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

32;

a light chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

39;

a light chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

46; and

a light chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

53;

(e) a heavy chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

19;

a heavy chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

26;

a heavy chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

33;

a light chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

40;

a light chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

47; and

a light chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

54;

(f) a heavy chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

20;

a heavy chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

27;

a heavy chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

34;

a light chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

41;

a light chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

48; and

a light chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

55;

or

(g) a heavy chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

21;

a heavy chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

28;

a heavy chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

35;

a light chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

42;

a light chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

49; and

a light chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

56.

11. The method of claim 10, wherein the anti-PD-1 monoclonal antibody or antigen-binding portion thereof comprises a heavy chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO:

18;

a heavy chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

25;

a heavy chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

32;

a light chain CDR1 comprising amino acids having the sequence set forth in SEQ ID NO;

39;

a light chain CDR2 comprising amino acids having the sequence set forth in SEQ ID NO;

46; and

a light chain CDR3 comprising amino acids having the sequence set forth in SEQ ID NO;

53.

12. The method of claim 11, wherein the anti-PD-1 monoclonal antibody or antigen-binding portion thereof comprises:

(a) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

1 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

8;

(b) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

2 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

9;

(c) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

3 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

10;

(d) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

4 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

11;

(e) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

5 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

12;

(f) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

6 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

13;

or

(g) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

7 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

14.

13. The method of claim 12, wherein the anti-PD-1 monoclonal antibody or antigen-binding portion thereof comprises a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO:

4 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO;

11.

34. The method of any one of claims 4, 8, 10 and 32, wherein the combination of the anti-PD-1 antibody or antigen-binding portion thereof and the anti-CTLA-4 antibody exhibits a synergistic therapeutic effect on cancer treatment.

32. A method for treating a cancer in a subject comprising administering to the subject a combination of an anti-PD-1 monoclonal antibody, or an antigen-binding portion thereof, and an anti-CTLA-4 monoclonal antibody designated 10D1, wherein the anti-PD-1 monoclonal antibody or antigen-binding portion thereof cross-competes for binding to human PD-1 with a reference antibody or antigen-binding portion thereof comprising a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO:

4 and a light chain variable region having the amino acid sequence set forth in SEQ ID NO;

11.

33. The method of claim 32, wherein the anti-PD-1 antibody comprises a heavy chain constant region that is of a human IgG4 isotype and a light chain constant region that is a human kappa constant region.

35. The method of claim 32, wherein the cancer is melanoma.

36. The method of claim 32, wherein the cancer is lung cancer.

37. The method of claim 32, wherein the cancer is renal cancer.