Anti-PD-1 antibodies and methods of use thereof
The present invention provides antagonizing antibodies that bind to programmed cell death protein 1 (PD-1) and methods of using same. The anti-PD-1 antibodies can be used therapeutically alone or in combination with other therapeutics to treat cancer and other diseases.
1. An isolated antagonist antibody that specifically binds to PD-1 and comprises:
a heavy chain variable region (VH) comprising a VII complementarity determining region one (CDR1), VII CDR2, and VH CDR3 of the VH having an amino acid sequence selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, and
a light chain variable region (VL) comprising a VL CDR1, VL CDR2, and VL CDR3 of the VL having an amino acid sequence selected from the group consisting of SEQ ID NO: 2, SEQ ID NO:7, SEQ ID NO: 8, and SEQ ID NO: 9.
2. The isolated antagonist antibody of claim 1 , wherein the antibody comprises a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 13, 14, or 15, a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 16, 17, 24, 25, 27, 28, 35, or 36, a VII CDR3 comprising the amino acid sequence shown in SEQ ID NO: 18, 23, 26, or 37, a VL CDR1 comprising the amino acid sequence shown in SEQ ID NO: 10, 22, 30, or 32, a VL CDR2 comprising the amino acid sequence shown in SEQ ID NO: 11, 20, or 33, and a VL CDR3 comprising the amino acid sequence shown in SEQ ID NO: 12, 21, 31, or 34.
3. The isolated antagonist antibody of claim 1 , wherein the antibody comprises a VH comprising the amino acid sequence shown in SEQ ID NO: 3, 4, 5, or 6, or a variant thereof with one or several conservative amino acid substitutions in residues that are not within a CDR.
4. The isolated antagonist antibody of claim 3 , wherein the antibody comprises a VL comprising the amino acid sequence shown in SEQ ID NO: 2, 7, 8, or 9, or a variant thereof with one or several amino acid substitutions in amino acids that are not within a CDR.
5. The isolated antagonist antibody of claim 1 , wherein the antibody comprises a VH comprising the amino acid sequence shown in SEQ ID NO: 3, 4, 5, or 6, and a VL comprising the amino acid sequence shown in SEQ ID NO: 2, 7, 8, or 9.
6. The isolated antagonist antibody of claim 1 , wherein the antibody comprises a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 29 or 38 and a light chain comprising the amino acid sequence shown in SEQ ID NO: 39.
7. The isolated antagonist antibody of claim 1 , wherein the antibody comprises a constant region.
8. The isolated antagonist antibody of claim 7 , wherein the antibody has an isotype that is selected from the group consisting of IgG 2 , IgG 2Δa , IgG 4 , IgG 4Δb , IgG 4Δc , IgG 4 S228P, IgG 4Δb S228P, and IgG 4Δc S228P.
9. The isolated antagonist antibody of claim 7 , wherein the constant region is IgG 4 S228P.
10. The isolated antagonist antibody of claim 1 , wherein each CDR of the antibody is defined in accordance with the Kabat definition, the Chothia definition, the combination of the Kabat definition and the Chothia definition, the AbM definition, or the contact definition of CDR.
11. An isolated anti-PD-1 antibody, wherein the antibody comprises a VII CDR1 comprising the amino acid sequence of SEQ ID NO: 13, a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 17, a VH CDR3 comprising the amino acid sequence shown in SEQ ID NO: 23, a VL CDR1 comprising the amino acid sequence shown in SEQ ID NO: 10, a VL CDR2 comprising the amino acid sequence shown in SEQ ID NO: 20, and a VL CDR3 comprising the amino acid sequence shown in SEQ ID NO: 21.
12. The isolated antibody of claim 1 , wherein the antibody (1) promotes IFNγ, (2) TNF secretion from T cells, or both (1) and (2).
13. The isolated antibody of claim 1 , wherein the antibody promotes proliferation of T cells.
14. The isolated antibody of claim 1 , wherein the antibody inhibits tumor growth in a subject in need thereof.
15. The isolated antibody of claim 1 , wherein the antibody binds human PD-1 and mouse PD-1.
16. The isolated antibody of claim 15 , wherein the antibody binds human PD-1 with an affinity of about 0.73 nM at 25° C. as measured by surface plasmon resonance.
17. An isolated cell line that produces the antibody of claim 1 .
18. An isolated nucleic acid encoding the antibody of claim 1 .
19. A recombinant expression vector comprising the nucleic acid of claim 18 .
20. A host cell comprising the expression vector of claim 19 .
21. A hybridoma capable of producing the antibody of claim 1 .
22. A method of producing an anti-PD-1 antagonist antibody, the method comprising: culturing a cell line that recombinantly produces the antibody of claim 1 under conditions wherein the antibody is produced; and recovering the antibody.
23. A method of producing an anti-PD-1 antagonist antibody, the method comprising: culturing a cell line comprising nucleic acid encoding an antibody comprising a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 29 or 38 and a light chain comprising the amino acid sequence shown in SEQ ID NO: 39 under conditions wherein the antibody is produced; and recovering the antibody.
24. The method of claim 23 , wherein the heavy and light chains of the antibody are encoded on separate vectors.
25. The method of claim 23 , wherein the heavy and light chains of the antibody are encoded on the same vector.
26. A pharmaceutical composition comprising the antibody of claim 1 , and a pharmaceutically acceptable carrier.
27. A kit for the treatment of cancer comprising the pharmaceutical composition of claim 26 .
28. The isolated anti-PD-1 antibody of claim 11 , wherein the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 4.
29. The isolated anti-PD-1 antibody of claim 28 , wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 29 or 38.
30. The isolated anti-PD-1 antibody of claim 11 , wherein the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 8.
31. The isolated anti-PD-1 antibody of claim 30 , wherein the antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 39.
32. An isolated cell line that produces the antibody of claim 11 .
33. A recombinant expression vector comprising a nucleic acid encoding the antibody of claim 11 .
34. A hybridoma capable of producing the antibody of claim 11 .
35. A pharmaceutical composition comprising the antibody of claim 11 and a pharmaceutically acceptable carrier.
36. An isolated antagonist antibody that specifically binds to PD-1, wherein the antibody comprises a heavy chain variable region (VII) comprising the amino acid sequence shown in SEQ ID NO: 4, and a light chain variable region (VL) comprising the amino acid sequence shown in SEQ ID NO: 8.
37. An isolated cell line that produces the antibody of claim 36 .
38. A recombinant expression vector comprising a nucleic acid encoding the antibody of claim 36 .
39. A hybridoma capable of producing the antibody of claim 36 .
40. A pharmaceutical composition comprising the antibody of claim 36 and a pharmaceutically acceptable carrier.
41. An isolated antagonist antibody that specifically binds to PD-1, wherein the antibody comprises a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 29 or 38 and a light chain comprising the amino acid sequence shown in SEQ ID NO: 39.
42. An isolated cell line that produces an antibody of claim 41 .
43. A recombinant expression vector comprising a nucleic acid encoding an antibody of claim 41 .
44. A hybridoma capable of producing an antibody of claim 41 .
45. A pharmaceutical composition comprising an antibody of claim 41 and a pharmaceutically acceptable carrier.