PD-1 BINDING PROTEINS

• US 20110008369A1
• Filed: 03/03/2009
• Published: 01/13/2011
• Est. Priority Date: 03/12/2008
• Status: Active Grant

• Current Assignee: Merck Sharp & Dohme LLC (Merck & Co., Inc.)
• Original Assignee: Merck Sharp & Dohme Corporation (Merck & Co., Inc.)
• Inventors: Freed, Daniel C., Casimiro, Danilo R., Tang, Aimin, Fu, Tong-Ming, Finnefrock, Adam C., Li, Fengsheng
• Granted Patent: US 8,168,757 B2
• Time in Patent Office: Days
• US Class Current: 424/172.1
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: C07K 16/2896 against molecules with a "CD"-designation, not provided for elsewhere
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• 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 features PD-1 binding proteins, a subset of which inhibits binding of PD-L1 to the PD-1 receptor. These binding proteins can be employed to modulate the immune system through the manipulation of the PD-1 signaling pathway, enhancing host immunity to treat infections and cancer.

Claims

1. An isolated PD-1 binding protein comprising a first variable region and a second variable region, wherein said binding protein specifically binds a human PD-1 epitope contained within a target region selected from the group consisting of a PD-L1-blocking target region and a PD-L1-non-blocking target region.

3. An isolated PD-1 binding protein of claim 1, wherein said target region is a PD-L1-blocking target region, comprising a first variable region and a second variable region, wherein said binding protein specifically binds a human PD-1 epitope contained within a PD-L1-blocking target region, wherein said first variable region is a heavy chain variable (V_h) region comprising at least one CDR selected from the group consisting of:

a first V_h CDR comprising either SEQ ID NO;

6, SEQ ID NO;

14, SEQ ID NO;

20, SEQ ID NO;

28, SEQ ID NO;

36, SEQ ID NO;

42 or SEQ ID NO;

49, or an amino acid sequence differing from SEQ ID NOs;

6, 14, 20, 28, 36, 42 or 49 by one amino acid;

a second V_h CDR comprising either SEQ ID NO;

7, SEQ ID NO;

15, SEQ ID NO;

21, SEQ ID NO;

29, SEQ ID NO;

37, SEQ ID NO;

43 or SEQ ID NO;

50, or an amino acid sequence differing from SEQ ID NOs;

7, 15, 21, 29, 37, 43 or 50 by one amino acid; and

,

a third V_h CDR comprising either SEQ ID NO;

8, SEQ ID NO;

16, SEQ ID NO;

22, SEQ ID NO;

30, SEQ ID NO;

38, SEQ ID NO;

44 or SEQ ID NO;

51, or an amino acid sequence differing from SEQ ID NOs;

8, 16, 22, 30, 38, 44 or 51 by one amino acid.

6. The binding protein of claim 3, wherein said first V_h CDR comprises SEQ ID NO:

6;

said second V_h CDR comprises SEQ ID NO;

7; and

, said third V_h CDR comprises SEQ ID NO;

8.

7. The binding protein of claim 3, wherein said second variable region is a light chain variable (V_l) region comprising at least one CDR selected from the group consisting of:

a first V_l CDR comprising either SEQ ID NO;

9, SEQ ID NO;

23, SEQ ID NO;

31, SEQ ID NO;

39, SEQ ID NO;

45 or SEQ ID NO;

52, or an amino acid sequence differing from SEQ ID NOs;

9, 23, 31, 39, 45 or 52 by one amino acid;

a second V_l CDR comprising either SEQ ID NO;

10, SEQ ID NO;

17, SEQ ID NO;

24, SEQ ID NO;

32 or SEQ ID NO;

53, or an amino acid sequence differing from SEQ ID NOs;

10, 17, 24, 32 or 53 by one amino acid; and

,

a third V_l CDR comprising either SEQ ID NO;

11, SEQ ID NO;

25, SEQ ID NO;

33, SEQ ID NO;

46 or SEQ ID NO;

54, or an amino acid sequence differing from SEQ ID NOs;

11, 25, 33, 46 or 54 by one amino acid.

12. The binding protein of claim 7, wherein said first V_l CDR comprises SEQ ID NO:

9;

said second V_l CDR comprises SEQ ID NO;

10; and

, said third V_l CDR comprises SEQ ID NO;

11.

15. The binding protein of claim 3, wherein the first variable region is a heavy chain variable (V_h) region comprising a first V_h CDR, a second V_h CDR and a third V_h CDR, and the second variable region is a light chain variable (V_l) region comprising a first V_l CDR, a second V_l CDR and a third V_l CDR, wherein said first V_h CDR, said second V_h CDR, said third V_h CDR, said first V_l CDR, said second V_l CDR and said third V_l CDR comprise the amino acid sequences selected from the group consisting of:

(a) SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

8, SEQ ID NO;

9, SEQ ID NO;

10 and SEQ ID NO;

11, respectively;

(b) SEQ ID NO;

14, SEQ ID NO;

15, SEQ ID NO;

16, SEQ ID NO;

9, SEQ ID NO;

17 and SEQ ID NO;

11, respectively;

(c) SEQ ID NO;

20, SEQ ID NO;

21, SEQ ID NO;

22, SEQ ID NO;

23, SEQ ID NO;

24 and SEQ ID NO;

25, respectively;

(d) SEQ ID NO;

42, SEQ ID NO;

43, SEQ ID NO;

44, SEQ ID NO;

45, SEQ ID NO;

24 and SEQ ID NO;

46, respectively;

(e) SEQ ID NO;

42, SEQ ID NO;

43, SEQ ID NO;

44, SEQ ID NO;

52, SEQ ID NO;

53 and SEQ ID NO;

54, respectively;

(f) SEQ ID NO;

49, SEQ ID NO;

50, SEQ ID NO;

51, SEQ ID NO;

45, SEQ ID NO;

24 and SEQ ID NO;

46, respectively;

(g) SEQ ID NO;

49, SEQ ID NO;

50, SEQ ID NO;

51, SEQ ID NO;

52, SEQ ID NO;

53 and SEQ ID NO;

54, respectively;

(h) SEQ ID NO;

28, SEQ ID NO;

29, SEQ ID NO;

30, SEQ ID NO;

31, SEQ ID NO;

32 and SEQ ID NO;

33, respectively; and

,

(i) SEQ ID NO;

36, SEQ ID NO;

37, SEQ ID NO;

38, SEQ ID NO;

39, SEQ ID NO;

24 and SEQ ID NO;

25, respectively.

16. The binding protein of claim 15, wherein said binding protein is an antibody.

17. The binding protein of claim 16, wherein

said V_h region comprises said first V_h CDR consisting of SEQ ID NO:

6, said second V_h CDR consisting of SEQ ID NO;

7, and said third V_h CDR consisting of SEQ ID NO;

8; and

,

said V_l region comprises said first V_l CDR consisting of SEQ ID NO;

9, said second V_l CDR consisting of SEQ ID NO;

10, and said third V_l CDR consisting of SEQ ID NO;

11.

18. The binding protein of claim 3, wherein said binding protein is an antibody comprising a first variable region which is a heavy chain variable (V_h) region and a second variable region which is a light chain variable (V_l) region, and wherein said V_h region comprises either:

(a) SEQ ID NO;

4 or a humanized SEQ ID NO;

4;

(b) SEQ ID NO;

12 or a humanized SEQ ID NO;

12;

(c) SEQ ID NO;

18 or a humanized SEQ ID NO;

18;

(d) SEQ ID NO;

40 or a humanized SEQ ID NO;

40;

(e) SEQ ID NO;

47 or a humanized SEQ ID NO;

47;

(f) SEQ ID NO;

26 or a humanized SEQ ID NO;

26;

or

(g) SEQ ID NO;

34 or a humanized SEQ ID NO;

34.

19. The binding protein of claim 18, wherein said V_h region comprises SEQ ID NO:

4.

21. The binding protein of claim 18, wherein said V_l region comprises SEQ ID NO:

5.

22. The binding protein of claim 18, wherein said antibody comprises either:

(a) a V_l region comprising SEQ ID NO;

5 or a humanized SEQ ID NO;

5, and a V_h region comprising SEQ ID NO;

4 or a humanized SEQ ID NO;

4;

(b) a V_l region comprising SEQ ID NO;

13 or a humanized SEQ ID NO;

13, and a V_h region comprising SEQ ID NO;

12 or a humanized SEQ ID NO;

12;

(c) a V_l region comprising SEQ ID NO;

19 or a humanized SEQ ID NO;

19, and a V_h region comprising SEQ ID NO;

18 or a humanized SEQ ID NO;

18;

(d) a V_l region comprising SEQ ID NO;

41 or a humanized SEQ ID NO;

41, and a V_h region comprising SEQ ID NO;

40 or a humanized SEQ ID NO;

40;

(e) a V_l region comprising SEQ ID NO;

41 or a humanized SEQ ID NO;

41, and a V_h region comprising SEQ ID NO;

47 or a humanized SEQ ID NO;

47;

(f) a V_l region comprising SEQ ID NO;

48 or a humanized SEQ ID NO;

48, and a V_h region comprising SEQ ID NO;

40 or a humanized SEQ ID NO;

40;

(g) a V_l region comprising SEQ ID NO;

48 or a humanized SEQ ID NO;

48, and a V_h region comprising SEQ ID NO;

47 or a humanized SEQ ID NO;

47;

(h) a V_l region comprising SEQ ID NO;

27 or a humanized SEQ ID NO;

27, and a V_h region comprising SEQ ID NO;

26 or a humanized SEQ ID NO;

26;

or,

(i) a V_l region comprising SEQ ID NO;

35 or a humanized SEQ ID NO;

35, and a V_h region comprising SEQ ID NO;

34 or a humanized SEQ ID NO;

34.

23. The binding protein of claim 22, wherein said V_h region comprises SEQ ID NO:

4 and said V_l region comprises SEQ ID NO;

5.

24. The binding protein of claim 22, wherein said antibody comprises a heavy chain comprising said V_h region and hinge, CH₁, CH₂ and CH₃ regions from an IgG₁, IgG₂, IgG₃ or IgG₄ subtype;

and a light chain comprising said V_l region and either a human kappa C_l or human lambda C_l.

26. An isolated PD-1 binding protein of claim 1, wherein said target region is a PD-L1-non-blocking target region, comprising a first variable region and a second variable region, wherein said binding protein specifically binds a human PD-1 epitope contained within a PD-L1-non-blocking target region, wherein said first variable region is a heavy chain variable (V_h) region comprising at least one CDR selected from the group consisting of:

a first V_h CDR comprising either SEQ ID NO;

57, SEQ ID NO;

64 or SEQ ID NO;

69, or an amino acid sequence differing from SEQ ID NOs;

57, 64 or 69 by one amino acid;

a second V_h CDR comprising either SEQ ID NO;

58, SEQ ID NO;

65 or SEQ ID NO;

70, or an amino acid sequence differing from SEQ ID NOs;

58, 65 or 70 by one amino acid; and

,

a third V_h CDR comprising either SEQ ID NO;

59, SEQ ID NO;

66 or SEQ ID NO;

71, or an amino acid sequence differing from SEQ ID NOs;

59, 66 or 71 by one amino acid.

28. The binding protein of claim 26, wherein said second variable region is a light chain variable (V_l) region comprising at least one CDR selected from the group consisting of:

a first V_l CDR comprising either SEQ ID NO;

60 or SEQ ID NO;

72, or an amino acid sequence differing from SEQ ID NOs;

60 or 72 by one amino acid;

a second V_l CDR comprising either SEQ ID NO;

61 or SEQ ID NO;

73, or an amino acid sequence differing from SEQ ID NOs;

61 or 73 by one amino acid; and

,

a third V_l CDR comprising either SEQ ID NO;

62 or SEQ ID NO;

74, or an amino acid sequence differing from SEQ ID NOs;

62 or 74 by one amino acid.

30. The binding protein of claim 26, wherein the first variable region is a heavy chain variable (V_h) region comprising a first V_h CDR, a second V_h CDR and a third V_h CDR, and the second variable region is a light chain variable (V_l) region comprising a first V_l CDR, a second V_l CDR and a third V_l CDR, wherein said first V_h CDR, said second V_h CDR, said third V_h CDR, said first V_l CDR, said second V_l CDR and said third V_l CDR comprise amino acid sequences selected from the group consisting of:

(a) SEQ ID NO;

57, SEQ ID NO;

58, SEQ ID NO;

59, SEQ ID NO;

60, SEQ ID NO;

61 and SEQ ID NO;

62, respectively; and

,

(b) SEQ ID NO;

69, SEQ ID NO;

70, SEQ ID NO;

71, SEQ ID NO;

72, SEQ ID NO;

73 and SEQ ID NO;

74, respectively.

31. The binding protein of claim 30, wherein said binding protein is an antibody.

39. A pharmaceutical composition comprising the a PD-1 binding protein selected from the group consisting of the proteins of claim 3 and the proteins of claim 26 and a pharmaceutically acceptable carrier.

40. A method of enhancing immune response of a subject comprising administering an effective amount of the pharmaceutical composition of claim 39.

2. (canceled)

4. (canceled)

5. (canceled)

8. (canceled)

9. (canceled)

10. (canceled)

11. (canceled)

13. (canceled)

14. (canceled)

20. (canceled)

25. (canceled)

32. The binding protein of claim 25, wherein said binding protein is an antibody comprising a first variable region which is a heavy chain variable (V_h) region and a second variable region which is a light chain variable (V_l) region, and wherein said V_h region comprises either:

(a) SEQ ID NO;

55 or a humanized SEQ ID NO;

55;

(b) SEQ ID NO;

63 or a humanized SEQ ID NO;

63;

or,

(c) SEQ ID NO;

67 or a humanized SEQ ID NO;

67.

33. The binding protein of claim 32, wherein said V_l region comprises either:

(a) SEQ ID NO;

56 or a humanized SEQ ID NO;

56;

or,

(b) SEQ ID NO;

68 or a humanized SEQ ID NO;

68.

34. The binding protein of claim 32, wherein said antibody comprises either:

(a) a V_l region comprising SEQ ID NO;

56 or a humanized SEQ ID NO;

56, and a V_h region comprising SEQ ID NO;

55 or a humanized SEQ ID NO;

55;

or,

(b) a V_l region comprising SEQ ID NO;

68 or a humanized SEQ ID NO;

68, and a V_h region comprising SEQ ID NO;

67 or a humanized SEQ ID NO;

67.

27. (canceled)

29. (canceled)

35. (canceled)

36. (canceled)

37. (canceled)

38. (canceled)