Anti-PD-1 antibodies and uses thereof

1. A method of treating cancer in a human subject comprising administering to the subject an effective amount of an anti-PD-1 antibody, wherein the anti-PD-1 antibody comprises (a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 21; (b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 22; (c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 23; (d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 25; (e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 26; and (f) LCDR3 comprising the amino acid sequence of SEQ ID NO: 27.

2. The method of claim 1 , wherein the cancer is selected from melanoma, non-small cell lung cancer (NSCLC), renal cell carcinoma (RCC), gastric cancer, bladder cancer, diffuse large B-cell lymphoma (DLBCL), Hodgkin's lymphoma, ovarian cancer, head & neck squamous cell cancer (HNSCC), mesothelioma, and triple negative breast cancer (TNBC).

3. The method of claim 2 , wherein the cancer is selected from melanoma, gastric cancer, head & neck squamous cell cancer (HNSCC), non-small cell lung cancer (NSCLC), and triple negative breast cancer (TNBC).

4. A method of enhancing an immune response in a human subject comprising administering to the subject an effective amount of an anti-PD-1 antibody, wherein the anti-PD-1 antibody comprises (a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 21; (b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 22; (c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 23; (d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 25;

(e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 26; and (f) LCDR3 comprising the amino acid sequence of SEQ ID NO: 27.

5. A method of increasing activation of a T cell in a human subject comprising administering to the subject an effective amount of an anti-PD-1 antibody, wherein the anti-PD-1 antibody comprises (a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 21; (b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 22; (c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 23; (d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 25;

(e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 26; and (f) LCDR3 comprising the amino acid sequence of SEQ ID NO: 27.

6. A method of reducing tumor size in a human subject with cancer comprising administering to the subject an effective amount of an anti-PD-1 antibody, wherein the anti-PD-1 antibody comprises (a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 21; (b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 22; (c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 23; (d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 25; (e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 26; and (f) LCDR3 comprising the amino acid sequence of SEQ ID NO: 27.

7. The method of claim 1 , wherein the subject is administered at least one additional therapeutic agent.

8. The method of claim 7 , wherein the additional therapeutic agent is administered concurrently or sequentially with the anti-PD-1 antibody.

9. The method of claim 7 wherein the additional therapeutic agent is selected from an anti-ICOS antibody and an anti-CTLA4 antibody.

10. The method of claim 9 , wherein the additional therapeutic agent is the anti-ICOS antibody.

11. A method of increasing the level of at least one cytokine selected from IFNyγ and IL-2 in a human subject, comprising administering to the subject an anti-PD-1 antibody, wherein the anti-PD-1 antibody comprises (a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 21; (b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 22; (c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 23; (d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 25; (e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 26; and (f) LCDR3 comprising the amino acid sequence of SEQ ID NO: 27.

12. The method of claim 11 , wherein the level of the cytokine is measured 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 8 hours, 10 hours, 12 hours, 14 hours, 16 hours, 18 hours, 20 hours, 22 hours, 24 hours, 36 hours, or 48 hours after administration of the anti-PD-1 antibody.

13. The method of claim 11 , wherein the at least one cytokine is IFNγ.

14. The method of claim 11 , wherein the at least one cytokine is IL-2.

15. The method of claim 11 , wherein the level of the cytokine is measured 24 hours after administration of the antibody.

16. The method of claim 11 , wherein the human subject has cancer.

17. The method of claim 16 , wherein the cancer is selected from melanoma, non-small cell lung cancer (NSCLC), renal cell carcinoma (RCC), gastric cancer, bladder cancer, diffuse large B-cell lymphoma (DLBCL), Hodgkin's lymphoma, ovarian cancer, head & neck squamous cell cancer (HNSCC), mesothelioma, and triple negative breast cancer (TNBC).

18. The method of claim 17 , wherein the cancer is selected from melanoma, gastric cancer, head & neck squamous cell cancer (HNSCC), non-small cell lung cancer (NSCLC), and triple negative breast cancer (TNBC).

19. The method of claim 11 , wherein an immune response is enhanced following administration of the anti-PD-1 antibody.

20. The method of claim 11 , wherein activation of T cells is increased following administration of the anti-PD-1 antibody.

21. The method of claim 16 , wherein tumor size is decreased following administration of the anti-PD-1 antibody.

22. The method of claim 11 , wherein the subject is administered at least one additional therapeutic agent.

23. The method of claim 22 , wherein the additional therapeutic agent is administered concurrently or sequentially with the anti-PD-1 antibody.

24. The method of claim 22 , wherein the additional therapeutic agent is selected from an anti-ICOS antibody and an anti-CTLA4 antibody.

25. The method of claim 24 , wherein the additional therapeutic is the anti-ICOS antibody.

26. The method of claim 1 , wherein a sample of the cancer has been determined to express PD-1.

27. The method of claim 26 , wherein the sample shows 1+, 2+, or 3+staining of PD-1 by immunohistochemistry (IHC).

28. The method of claim 1 , wherein the sample has been determined to have an elevated level of PD-Ll.

29. The method of claim 28 , wherein PD-L1 levels are determined using IHC.

30. The method of claim 1 , wherein the anti-PD-1 antibody comprises a heavy chain variable region (V H ) that is at least 90% identical to the amino acid sequence of SEQ ID NO: 20 and a light chain variable region (V L ) that is at least 90% identical to the amino acid sequence of SEQ ID NO: 24.

31. The method of claim 1 , wherein the anti-PD-1 antibody comprises a V H comprising the amino acid sequence of SEQ ID NO: 20 and a V L comprising the amino acid sequence of SEQ ID NO: 24.

32. The method of claim 1 , wherein the anti-PD-1 antibody binds to human PD-1 with an affinity (K D ) of less than 5 nM.

33. The method of claim 32 , wherein affinity is determined using biolayer interferometry.

34. The method of claim 1 , wherein the anti-PD-1 antibody is an antibody fragment selected from a Fab, Fab′, Fv, scFv or (Fab′)2 fragment.

35. The method of claim 1 , wherein the anti-PD-1 antibody is a full length antibody.

36. The method of claim 4 , wherein the anti-PD-1 antibody comprises a heavy chain variable region (V H ) that is at least 90% identical to the amino acid sequence of SEQ ID NO: 20 and a light chain variable region (V L ) that is at least 90% identical to the amino acid sequence of SEQ ID NO: 24.

37. The method of claim 4 , wherein the anti-PD-1 antibody comprises a V H comprising the amino acid sequence of SEQ ID NO: 20 and a V L comprising the amino acid sequence of SEQ ID NO: 24.

38. The method of claim 4 , wherein the anti-PD-1 antibody binds to human PD-1 with an affinity (KD) of less than 5 nM.

39. The method of claim 38 , wherein affinity is determined using biolayer interferometry.

40. The method of claim 4 , wherein the anti-PD-1 antibody is an antibody fragment selected from a Fab, Fab′, Fv, scFv or (Fab′)2 fragment.

41. The method of claim 4 , wherein the anti-PD-1 antibody is a full length antibody.

42. The method of claim 5 , wherein the anti-PD-1 antibody comprises a heavy chain variable region (V H ) that is at least 90% identical to the amino acid sequence of SEQ ID NO: 20 and a light chain variable region (V L ) that is at least 90% identical to the amino acid sequence of SEQ ID NO: 24.

43. The method of claim 5 , wherein the anti-PD-1 antibody comprises a V H comprising the amino acid sequence of SEQ ID NO: 20 and a V L comprising the amino acid sequence of SEQ ID NO: 24.

44. The method of claim 5 , wherein the anti-PD-1 antibody binds to human PD-1 with an affinity (KD) of less than 5 nM.

45. The method of claim 44 , wherein affinity is determined using biolayer interferometry.

46. The method of claim 5 , wherein the anti-PD-1 antibody is an antibody fragment selected from a Fab, Fab′, Fv, scFv or (Fab′)2 fragment.

47. The method of claim 5 , wherein the anti-PD-1 antibody is a full length antibody.

48. The method of claim 6 , wherein the anti-PD-1 antibody comprises a heavy chain variable region (V H ) that is at least 90% identical to the amino acid sequence of SEQ ID NO: 20 and a light chain variable region (V L ) that is at least 90% identical to the amino acid sequence of SEQ ID NO: 24.

49. The method of claim 6 , wherein the anti-PD-1 antibody comprises a V H comprising the amino acid sequence of SEQ ID NO: 20 and a V L comprising the amino acid sequence of SEQ ID NO: 24.

50. The method of claim 6 , wherein the anti-PD-1 antibody binds to human PD-1 with an affinity (KD) of less than 5 nM.

51. The method of claim 50 , wherein affinity is determined using biolayer interferometry.

52. The method of claim 6 , wherein the anti-PD-1 antibody is an antibody fragment selected from a Fab, Fab′, Fv, scFv or (Fab′)2 fragment.

53. The method of claim 6 , wherein the anti-PD-1 antibody is a full length antibody.

54. The method of claim 11 , wherein the anti-PD-1 antibody comprises a heavy chain variable region (V H ) that is at least 90% identical to the amino acid sequence of SEQ ID NO: 20 and a light chain variable region (V L ) that is at least 90% identical to the amino acid sequence of SEQ ID NO: 24.

55. The method of claim 11 , wherein the anti-PD-1 antibody comprises a V H comprising the amino acid sequence of SEQ ID NO: 20 and a V L comprising the amino acid sequence of SEQ ID NO: 24.

56. The method of claim 11 , wherein the anti-PD-1 antibody binds to human PD-1 with an affinity (KD) of less than 5 nM.

57. The method of claim 56 , wherein affinity is determined using biolayer interferometry.

58. The method of claim 11 , wherein the anti-PD-1 antibody is an antibody fragment selected from a Fab, Fab′, Fv, scFv or (Fab′)2 fragment.

59. The method of claim 11 , wherein the anti-PD-1 antibody is a full length antibody.

60. A method of stimulating the activity of an immune cell or reducing the downmodulation of an immune cell in a human subject comprising administering to the subject an effective amount of an anti-PD-1 antibody, wherein the anti-PD-1 antibody comprises (a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 21; (b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 22; (c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 23; (d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 25; (e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 26; and (f) LCDR3 comprising the amino acid sequence of SEQ ID NO: 27.

61. The method of claim 60 , wherein the anti-PD-1 antibody comprises a heavy chain variable region (V H ) that is at least 90% identical to the amino acid sequence of SEQ ID NO: 20 and a light chain variable region (V L ) that is at least 90% identical to the amino acid sequence of SEQ ID NO: 24.

62. The method of claim 60 , wherein the anti-PD-1 antibody comprises a V H comprising the amino acid sequence of SEQ ID NO: 20 and a V L comprising the amino acid sequence of SEQ ID NO: 24.

63. The method of claim 60 , wherein the anti-PD-1 antibody binds to human PD-1 with an affinity (K D ) of less than 5 nM.

64. The method of claim 63 , wherein affinity is determined using biolayer interferometry.

65. The method of claim 60 , wherein the anti-PD-1 antibody is an antibody fragment selected from a Fab, Fab′, Fv, scFv or (Fab′)2 fragment.

66. The method of claim 60 , wherein the anti-PD-1 antibody is a full length antibody.

67. The method of claim 31 , wherein the anti-PD-1 antibody is a full length antibody.

68. The method of claim 37 , wherein the anti-PD-1 antibody is a full length antibody.

69. The method of claim 43 , wherein the anti-PD-1 antibody is a full length antibody.

70. The method of claim 49 , wherein the anti-PD-1 antibody is a full length antibody.

71. The method of claim 55 , wherein the anti-PD-1 antibody is a full length antibody.

72. The method of claim 62 , wherein the anti-PD-1 antibody is a full length antibody.