Anti-PD-L1 antibodies, compositions and articles of manufacture
The present application relates to anti-PD-L1 antibodies, nucleic acid encoding the same, therapeutic compositions thereof, and their use enhance T-cell function to upregulate cell-mediated immune responses and for the treatment of T cell dysfunctional disorders, including infection (e.g., acute and chronic) and tumor immunity.
1. An isolated heavy chain variable region polypeptide that specifically binds to PD-L1 comprising an HVR-H1, HVR-H2 and HVR-H3 sequence, wherein:
(a)
(SEQ ID NO: 1)
the HVR-H1 sequence is GFTFSX 1 SWIH;
(b)
(SEQ ID NO: 2)
the HVR-H2 sequence is AWIX 2 PYGGSX 3 YYADSVKG;
(c)
(SEQ ID NO: 3)
the HVR-H3 sequence is RHWPGGFDY;
further wherein: X 1 is D or G; X 2 is S or L; X 3 is T or S.
2. The polypeptide of claim 1 wherein X 1 is D; X 2 is S and X 3 is T.
3. The polypeptide of claim 1 further comprising variable region heavy chain framework sequences juxtaposed between the HVRs according to the formula: (HC-FR1)-(HVR-H1)-(HC-FR2)-(HVR-H2)-(HC-FR3)-(HVR-H3)-(HC-FR4).
4. The polypeptide of claim 3 wherein the framework sequences are human.
5. The polypeptide of claim 4 wherein the framework sequences are VH subgroup III consensus framework.
6. The polypeptide of claim 5 wherein one or more of the framework sequences is the following:
(SEQ ID NO: 4)
HC-FR1 is EVQLVESGGGLVQPGGSLRLSCAAS
(SEQ ID NO: 5)
HC-FR2 is WVRQAPGKGLEWV
(SEQ ID NO: 6)
HC-FR3 is RETISADTSKNTAYLQMNSLRAEDTAVYYCAR
(SEQ ID NO: 7)
HC-FR4 is WGQGTLVTVSA.
7. The isolated heavy chain polypeptide of claim 1 in combination with a variable region light chain comprising an HVR-L1, HVR-L2 and HVR-L3, wherein:
(a)
(SEQ ID NOs: 8)
the HVR-L1 sequence is RASQX 4 X 5 X 6 TX 7 X 8 A;
(b)
(SEQ ID NOs: 9)
the HVR-L2 sequence is SASX 9 LX 10 S,;
and
(SEQ ID NOs: 10)
(c)
the HVR-L3 sequence is QQX 11 X 12 X 13 X 14 PX 15 T;
further wherein: X 4 is D or V; X 5 is V or I; X 6 is S or N; X 7 is A or F; X 8 is V or L; X 9 is F or T; X 10 is Y or A; X 11 is Y, G, F, or S; X 12 , is L, Y, F or W; X 13 is Y, N, A, T, G, F or I; X 14 is H, V, P, T or I; X 15 is A, W, R, P or T.
8. The polypeptide of claim 7 wherein X 4 is D; X 5 is V; X 6 is S; X 7 is A; X 8 is V; X 9 is F; X 10 is Y; X 11 is Y; X 12 is L; X 13 is Y; X 14 is H; X 15 is A.
9. The polypeptide of claim 7 further comprising variable region light chain framework sequences juxtaposed between the HVRs according to the formula: (LC-FR1)-(HVR-L1)-(LC-FR2)-(HVR-L2)-(LC-FR3)-(HVR-L3)-(LC-FR4).
10. The polypeptide of claim 9 wherein the framework sequences are, human.
11. The polypeptide of claim 10 wherein the framework sequences are VL kappa I consensus framework.
12. The polypeptide of claim 11 wherein one or more of the framework sequences is the following:
(SEQ ID NO: 11)
LC-FR1 is DIQMTQSPSSLSASVGDRVTITC;
(SEQ ID NO: 12)
LC-FR2 is WYQQKPGKAPKLLIY;
(SEQ ID NO: 13)
LC-FR3 is GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC;
(SEQ ID NO: 14)
LC-FR4 is FGQGTKVEIKR.
13. An isolated anti-PD-L1 antibody or antigen binding fragment thereof, wherein the antibody or antibody fragment comprises a heavy chain and a light chain variable region sequence, wherein:
(a) the heavy chain comprises an HVR-H1, HVR-H2 and HVR-H3, wherein further:
(i)
(SEQ ID NO: 1)
the HVR-H1 sequence is GFTFSX 1 SWIH;
(ii)
(SEQ ID NO: 2)
the HVR-H2 sequence is AWIX 2 PYGGSX 3 YYADSVKG;
(iii)
(SEQ ID NO: 3)
the HVR-H3 sequence is RHWPGGFDY,;
and
(b) the light chain comprises an HVR-L1, HVR-L2 and HVR-L3, wherein further:
(iv)
(SEQ ID NOs: 8)
the HVR-L1 sequence is RASQX 4 X 5 X 6 TX 7 X 8 A;
(v)
(SEQ ID NOs: 9)
the HVR-L2 sequence is SASX 9 LX 10 S;
(vi)
(SEQ ID NOs: 10)
the HVR-L3 sequence is QQX 11 X 12 X 13 X 14 PX 15 T;
wherein: X 1 is D or G; X 2 is S or L; X 3 is T or S; X 4 may be D or V; X 5 may be V or I; X 6 may be S or N; X 7 may be A or F; X 8 may be V or L; X 9 may be F or T; X 10 may be Y or A; X 11 may be Y, G, F, or S; X 12 may be L, Y, F or W; X 13 may be Y, N, A, T, G, F or I; X 14 may be H, V, P, T or I; X 15 may be A, W, R, P or T.
14. The antibody or antibody fragment of claim 13 wherein X 1 is D; X 2 is S and X 3 is T.
15. The antibody or antibody fragment of claim 13 , wherein X 4 =D, X 5 =V, X 6 =S, X 7 =A and X 8 =V, X 9 =F, and X 10 =Y, X 11 =Y, X 12 =L, X 13 =Y, X 14 =H and X 15 =A.
16. The antibody or antibody fragment of claim 13 , wherein X 1 =D, X 2 =S and X 3 =T, X 4 =D, X 5 =V, X 6 =S, X 7 =A and X 8 =V, X 9 =F, and X 10 =Y, X 11 =Y, X 12 =L, X 13 =Y, X 14 =H and X 15 =A.
17. The antibody or antibody fragment of claim 14 further wherein the antibody or antibody fragment comprises:
variable region heavy chain framework sequences juxtaposed between the HVRs according to the formula: (HC-FR1)-(HVR-H1)-(HC-FR2)-(HVR-H2)-(HC-FR3)-(HVR-H3)-(HC-FR4).
18. The antibody or antibody fragment of claim 17 wherein the framework sequences are human.
19. The antibody or antibody fragment of claim 18 wherein the variable region heavy chain framework sequences are VH subgroup III consensus framework.
20. The antibody or antibody fragment of claim 19 wherein one or more of the framework sequences is the following:
(SEQ ID NO: 4)
HC-FR1 is EVQLVESGGGLVQPGGSLRLSCAAS;
(SEQ ID NO: 5)
HC-FR2 is WVRQAPGKGLEWV;
(SEQ ID NO: 6)
HC-FR3 is RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR;
(SEQ ID NO: 7)
HC-FR4 is WGQGTLVTVSA.
21. The antibody or antibody fragment of claim 15 further wherein the antibody comprises:
variable region light chain framework sequences juxtaposed between the HVRs according to the formula: (LC-FR1)-(HVR-L1)-(LC-FR2)-(HVR-L2)-(LC-FR3)-(HVR-L3)-(LC-FR4).
22. The antibody or antibody fragment of claim 21 wherein the variable region light chain framework sequences are VL kappa I consensus framework.
23. The antibody or antibody fragment of claim 22 wherein one or more of the framework sequences is the following:
(SEQ ID NO: 11)
LC-FR1 is DIQMTQSPSSLSASVGDRVTITC;
(SEQ ID NO: 12)
LC-FR2 is WYQQKPGKAPKLLIY;
(SEQ ID NO: 13)
LC-FR3 is GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC;
and
(SEQ ID NO: 14)
LC-FR4 is FGQGTKVEIKR.
24. The antibody of or antibody fragment of claim 16 wherein the antibody or antibody fragment comprises:
(a) variable region heavy chain framework sequences juxtaposed between the HVRs according to the formula: (HC-FR1)-(HVR-H1)-(HC-FR2)-(HVR-H2)-(HC-FR3)-(HVR-H3)-(HC-FR4), and
(b) variable region light chain framework sequences juxtaposed between the HVRs according to the formula: (LC-FR1)-(HVR-L1)-(LC-FR2)-(HVR-L2)-(LC-FR3)-(HVR-L3)-(LC-FR4);
wherein the variable heavy chain framework sequences are the following:
(SEQ ID NO: 4)
(i) HC-FR1 is EVQLVESGGGLVQPGGSLRLSCAAS;
(SEQ ID NO: 5)
(ii) HC-FR2 is WVRQAPGKGLEWV;
(SEQ ID NO: 6)
(iii) HC-FR3 is RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR;
(SEQ ID NO: 7)
(iv) HC-FR4 is WGQGTLVTVSA;
and
further wherein the variable light chain framework sequences are the following:
(SEQ ID NO: 11)
(i) LC-FR1 is DIQMTQSPSSLSASVGDRVTITC;
(SEQ ID NO: 12)
(ii) LC-FR2 is WYQQKPGKAPKLLIY;
(SEQ ID NO: 13)
(iii) LC-FR3 is GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC;
(SEQ ID NO: 14)
(iv) LC-FR4 is FGQGTKVEIKR.
25. The antibody or antibody fragment of claim 24 further comprising a human constant region.
26. The antibody or antibody fragment of claim 25 , wherein the constant region is selected from the group consisting of IgG1, IgG2, IgG3 and IgG4.
27. The antibody of antibody fragment of claim 26 wherein the constant region is IgG1.
28. The antibody or antibody fragment of claim 24 , further comprising murine constant region.
29. The antibody or antibody fragment of claim 28 wherein the constant region is selected from the group consisting of IgG1, IgG2A, IgG2B and IgG3.
30. The antibody or antibody fragment of claim 29 , wherein the constant region is IgG2A.
31. The antibody or antibody fragment of claim 26 having reduced or minimal effector function.
32. The antibody or antibody fragment of claim 31 , wherein the minimal effector function results from an effector-less Fc mutation.
33. The antibody or antibody fragment of claim 32 , wherein the effector-less Fc mutation is N297A.
34. The antibody or antibody fragment of claim 32 , wherein the effector-less Fc mutation is D265A/N297A.
35. The antibody or antibody fragment of claim 31 , wherein the minimal effector function results from aglycosylation.
36. An isolated anti-PD-L1 antibody or antigen binding fragment thereof, wherein the antibody or antibody fragment comprises a heavy chain and light chain variable region sequence, wherein:
(a) the heavy chain comprises the sequence: EVQLVESGGGLVQPGGSLRLS CAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTI SADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVS A (SEQ ID NO:20), and
(b) the light chain comprises the sequence: DIQMTQSPSSLSASVGDRVTITC RASQDVSTAVAWYQQKPGKAPKWYSASFLYSGVPSRFSGSGSGTDFTL TISSLQPEDFATYYCQQYLYH PATFGQGTKVEIKR (SEQ ID NO:21).
37. The antibody or antibody fragment of claim 29 having reduced or minimal effector function.
38. The antibody or antibody fragment of claim 37 , wherein the minimal effector function results from an effector-less Fc mutation.
39. The antibody or antibody fragment of claim 38 , wherein the effector-less Fc mutation is N297A.
40. The antibody or antibody fragment of claim 38 , wherein the effector-less Fc mutation is D265A/N297A.
41. The antibody or antibody fragment of claim 37 , wherein the minimal effector function results from aglycosylation.
42. The antibody or antibody fragment of claim 36 further comprising a human constant region.
43. The antibody or antibody fragment of claim 42 , wherein the constant region is selected from the group consisting of IgG1, IgG2, IgG3 and IgG4.
44. The antibody or antibody fragment of claim 43 , wherein the constant region is IgG1.
45. The antibody or antibody fragment of claim 44 having reduced or minimal effector function.
46. The antibody or antibody fragment of claim 45 , wherein the minimal effector function results from an effector-less Fc mutation.
47. The antibody or antibody fragment of claim 46 , wherein the effector-less Fc mutation is N297A.
48. The antibody or antibody fragment of claim 46 , wherein the effector-less Fc mutation is D265A/N297A.
49. The antibody or antibody fragment of claim 45 , wherein the minimal effector function results from aglycosylation.
50. A composition comprising the anti-PD-L1 antibody or antigen binding fragment of any of claim 13 - 35 , 36 or 21 - 49 and at least one pharmaceutically-acceptable carrier.
51. An article of manufacture comprising the composition of claim 50 and at least one chemotherapeutic agent.
52. The article of manufacture according to claim 51 , wherein the chemotherapeutic agent is gemcitabine.
53. An article of manufacture comprising the composition of claim 50 , and at least one B7-family costimulatory molecule.
54. An article of manufacture comprising the composition of claim 50 and at least one antibiotic.
55. The article of manufacture according to claim 54 , wherein the antibiotic is an anti-viral agent.
56. The article of manufacture according to claim 55 , wherein anti-viral agent is a reverse transcriptase inhibitor.
57. The article of manufacture according to claim 56 , wherein the reverse transcriptase inhibitor is a polymerase inhibitor.
58. The article of manufacture according to claim 55 , wherein the anti-viral agent is a protease inhibitor.
59. An article of manufacture comprising the composition of claim 50 and at least one vaccine.
60. An article of manufacture comprising the composition of claim 50 .
61. The article of manufacture of claim 51 , wherein the chemotherapeutic agent is an anti-VEGF antibody.