Anti-PD-L1 antibodies and uses thereof

1. An isolated anti-PD-L1 antibody or antigen binding fragment thereof comprising:

a light chain variable region; and

a heavy chain variable region comprising HVR-H1, HVR-H2, and HVR-H3 sequences, wherein:

(a) the HVR-H1 sequence is X 1 YX 2 MX 3 (SEQ ID NO:1);

(b) the HVR-H2 sequence is SIYPSGGX 4 TFYADX 5 VKG (SEQ ID NO:2); and

(c) the HVR-H3 sequence is IKLGTVTTVX 6 Y (SEQ ID NO:3);

and further wherein:

X 1 is K, R, T, Q, G, A, W, M, I or S;

X 2 is V, R, K, L, M or I;

X 3 is H, T, N, Q, A, V, Y, W, F or M;

X 4 is F or I; X 5 is S or T; and

X 6 is E or D.

2. The isolated anti-PD-L1 antibody or antigen binding fragment of claim 1 , wherein:

(a) X 1 is M, I or S; X 2 is R, K, L, M or I; X 3 is F or M; X 4 is F or I; X 5 is S or T; and X 6 is E or D;

(b) X 1 is M, I or S; X 2 is L, M or I; X 3 is F or M; X 4 is I; X 5 is S or T; and X 6 is D; or

(c) X 1 is S; X 2 is I; X 3 is M; X 4 is I; X 5 is T; and X 6 is D.

3. The isolated anti-PD-L1 antibody or antigen binding fragment of claim 1 , wherein the heavy chain variable region comprises heavy chain framework sequences HC-FR1, HC-FR2, HC-FR3 and HC-FR4 interposed between the HVRs, thus forming a sequence of the formula: (HC-FR1)-(HVR-H1)-(HC-FR2)-(HVR-H2)-(HC-FR3)-(HVR-H3)-(HC-FR4).

4. The isolated anti-PD-L1 antibody or antigen binding fragment of claim 3 , wherein the heavy chain framework sequences are derived from human consensus heavy chain framework sequences or from human germline heavy chain framework sequences.

5. The isolated anti-PD-L1 antibody or antigen binding fragment of claim 3 , wherein one or more of the heavy chain framework sequences is selected from:

(a) HC-FR1 is

(SEQ ID NO: 4)

EVQLLESGGGLVQPGGSLRLSCAASGFTFS;

(b) HC-FR2 is 

(SEQ ID NO: 5)

WVRQAPGKGLEWVS;

(c) HC-FR3 is 

(SEQ ID NO: 6)

RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR; or

(d) HC-FR4 is

(SEQ ID NO: 7)

WGQGTLVTVSS.

6. The isolated anti-PD-L1 antibody or antigen binding fragment of claim 5 , further comprising a C H 1 domain or a C H 1, a C H 2 and a C H 3 domain.

7. The isolated anti-PD-L1 antibody or antigen binding fragment of claim 1 , wherein the light chain variable region comprises HVR-L1, HVR-L2 and HVR-L3 sequences, wherein:

(a) the HVR-L1 sequence is TGTX 7 X 8 DVGX 9 YNYVS (SEQ ID NO: 8);

(b) the HVR-L2 sequence is X 10 VX 11 X 12 RPS (SEQ ID NO: 9); and

(c) the HVR-L3 sequence is SSX 13 TX 14 X 15 X 16 X 17 RV (SEQ ID NO: 10);

and further wherein:

X 7 is N or S;

X 8 is T, R or S;

X 9 is A or G;

X 10 is E or D;

X 11 is I, N or S;

X 12 is D, H or N;

X 13 is F or Y;

X 14 is N or S;

X 15 is R, T or S;

X 16 is G or S; and

X 17 is I or T.

8. The isolated anti-PD-L1 antibody or antigen binding fragment of claim 7 , wherein:

(a) X 7 is N or S; X 8 is T, R or S; X 9 is A or G; X 10 is E or D; X II is N or S; X 12 is NT; X 13 is F or Y; X 14 is S; X 15 is S; X 16 is G or S; and X 17 is T; or

(b) X 7 is S; X 8 is S; X 9 is G; X 10 is D; X 11 is S; X 12 is N; X 13 is Y; X 14 is S; X 15 is S; X 16 is S; and X 17 is T.

9. The isolated anti-PD-L1 antibody or antigen binding fragment of claim 7 , wherein the light chain variable region comprises light chain framework sequences LC-FR1, LC-FR2, LC-FR3 and LC-FR4, interposed between the HVRs, thus forming a sequence of the formula: (LC-FR1)-(HVR-L1)-(LC-FR2)-(HVR-L2)-(LC-FR3)-(HVR-L3)-(LC-FR4).

10. The isolated anti-PD-L1 antibody or antigen binding fragment of claim 9 , wherein the light chain framework sequences are derived from human consensus light chain framework sequences or human germline light chain framework sequences.

11. The isolated anti-PD-L1 antibody or antigen binding fragment of claim 9 , wherein one or more of the light chain framework sequences is selected from:

(a) LC-FR1 is

(SEQ ID NO: 11)

QSALTQPASVSGSPGQSITISC;

(b) LC-FR2 is

(SEQ ID NO: 12)

WYQQHPGKAPKLMIY;

(c) LC-FR3 is

(SEQ ID NO: 13)

GVSNRFSGSKSGNTASLTISGLQAEDEADYYC; or

(d) LC-FR4 is

(SEQ ID NO: 14)

FGTGTKVTVL.

12. The isolated anti-PD-L1 antibody or antigen binding fragment of claim 11 , further comprising a C L domain.

13. The isolated anti-PD-L1 antibody or antigen binding fragment of claim 7 further comprising a human or murine constant region.

14. The isolated anti-PD-L1 antibody or antigen binding fragment of claim 13 , wherein the constant region is selected from the group consisting of IgG1, IgG2, IgG3 and IgG4.

15. An isolated anti-PD-L1 antibody or antigen binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein:

(a) the heavy chain variable region comprises an HVR-H1, an HVR-H2, and an HVR-H3, having at least 80% overall sequence identity to SYIMM (SEQ ID NO: 15), SIYPSGGITFYADTVKG (SEQ ID NO: 16), and IKLGTVTTVDY (SEQ ID NO: 17), respectively; and

(b) the light chain variable region comprises an HVR-L1, an HVR-L2, and an HVR-L3, having at least 80% overall sequence identity to TGTSSDVGGYNYVS (SEQ ID NO: 18), DVSNRPS (SEQ ID NO: 19) and SSYTSSSTRV (SEQ ID NO: 20), respectively.

16. The isolated anti-PD-L1 antibody or antigen binding fragment of claim 15 , wherein the heavy chain variable region or the light chain variable region sequence identity is at least 90%.

17. The isolated anti-PD-L1 antibody or antigen binding fragment of claim 15 , wherein, as compared to the sequences of HVR-H1 (SEQ ID NO: 15), HVR-H2 (SEQ ID NO: 16), and HVR-H3 (SEQ ID NO: 17), at least those amino acids that remain unchanged are as follows:

(a) residues 2 and 4 of SEQ ID NO:15,

(b) residues 1-7, 9-13, and 15-17 of SEQ ID NO:16, and

(c) residues 1-9 and 11 of SEQ ID NO:17;

and further wherein, as compared to the sequences of HVR-L1 (SEQ ID NO: 18), HVR-L2 (SEQ ID NO: 19) and HVR-L3 (SEQ ID NO: 20), at least those amino acids that remain unchanged are as follows:

(a) residues 1-3, 6-8, and 10-14 of SEQ ID NO:18;

(b) residues 1 and 5-7 of SEQ ID NO:19; and

(c) residues 1-2 and 9-10 of SEQ ID NO:20.

18. An isolated anti-PD-L1 antibody or antigen binding fragment thereof comprising a heavy chain variable region sequence and a light chain variable region sequence, wherein:

(a) the heavy chain variable region sequence has at least 85% sequence identity to the heavy chain variable region sequence:

(SEQ ID NO: 24)

EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYIMMWVRQAPGK

GLEWVSSIYPSGGITFYADTVKGRFTISRDNSKNTLYLQMNSL

RAEDTAVYYCARIKLGTVTTVDYWGQGTLVTVSS;

and

(b) the light chain variable region sequence has at least 85% sequence identity to the light chain variable region sequence:

(SEQ ID NO: 25)

QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPG

KAPKLMIYDVSNRPSGVSNRFSGSKSGNTASLTISGLQAEDEA

DYYCSSYTSSSTRVFGTGTKVTVL.

19. The isolated anti-PD-L1 antibody or antigen binding fragment of claim 18 , wherein the sequence identity of the heavy chain variable region or the light chain variable region is at least 90%.

20. The isolated anti-PD-L1 antibody or antigen binding fragment of claim 18 , wherein:

(a) the heavy chain variable region comprises the sequence:

(SEQ ID NO: 24)

EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYIMMWVRQAPGK

GLEWVSSIYPSGGITFYADTVKGRFTISRDNSKNTLYLQMNSL

RAEDTAVYYCARIKLGTVTTVDYWGQGTLVTVSS;

and

(b) the light chain variable region comprises the sequence:

(SEQ ID NO: 25)

QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHP

GKAPKLMIYDVSNRPSGVSNRFSGSKSGNTASLTISGLQAED

EADYYCSSYTSSSTRVFGTGTKVTVL.

21. The anti-PD-L1 antibody or antigen binding fragment of claim 20 , wherein the antibody further comprises a C H 1, a C H 2, a C H 3 and a C L domain.

22. The anti-PD-L1 antibody or antigen binding fragment of claim 7 , wherein the antibody binds to human, mouse or cynomolgus monkey PD-L1, or wherein the antibody is capable of blocking the interaction between human, mouse, or cynomolgus monkey PD-L1 and the respective human, mouse, or cynomolgus monkey PD-1 receptors.

23. The anti-PD-L1 antibody or antigen binding fragment of claim 7 , wherein the antibody binds to human PD-L1 with a K D of 5×10 −9 M or less.

24. An isolated anti-PD-L1 antibody or antigen binding fragment thereof wherein the antibody cross-competes for binding to PD-L1 with the anti-PD-L1 antibody or antigen binding fragment of claim 7 .

25. A composition comprising the anti-PD-L1 antibody or antigen binding fragment of claim 7 and at least one pharmaceutically acceptable carrier.

26. An isolated nucleic acid encoding the light chain or a heavy chain sequence of the anti-PD-L1 antibody or antigen binding fragment of claim 7 .

27. A vector comprising the nucleic acid of claim 26 .

28. A cultured host cell comprising the vector of claim 27 .

29. A process for making an anti-PD-L1 antibody or antigen binding fragment thereof comprising culturing the host cell of claim 28 under conditions suitable for the expression of the vector encoding the anti-PD-L1 antibody or antigen binding fragment, and recovering the antibody or fragment.

30. A kit of parts comprising the composition of claim 25 and at least one further therapeutic agent or vaccine.

31. A method of treating tumor immunity or cancer comprising administering to a subject in need thereof an effective amount of the composition of claim 25 .

32. An isolated anti PD-L1 antibody wherein the antibody comprises:

(a) a heavy chain variable region (VH) comprising an HVR-H1, HVR-H2 and HVR-H3 of SEQ ID NO: 32; and

(b) a light chain variable region (VL) comprising an HVR-L1, HVR-L2 and HVR-L3 of SEQ ID NO: 33.

33. The isolated anti PD-L1 antibody of claim 32 , wherein the HVR-H1 comprises the amino acid sequence of SEQ ID NO: 15, the HVR-H2 comprises the amino acid sequence of SEQ ID NO: 16, the HVR-H3 comprises the amino acid sequence of SEQ ID NO: 17, the HVR-L1 comprises the amino acid sequence of SEQ ID NO: 18, the HVR-L2 comprises the amino acid sequence of SEQ ID NO: 19, and the HVR-L3 comprises the amino acid sequence of SEQ ID NO: 20.

34. The isolated anti PD-L1 antibody of claim 32 , wherein the antibody comprises:

(a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 32, or the amino acid sequence of SEQ ID NO: 32 without the C-terminal lysine; and

(b) a light chain comprising the amino acid sequence of SEQ ID NO: 33.

35. The isolated anti PD-L1 antibody of claim 32 , wherein each HVR is defined in accordance with the Kabat definition, the Chothia definition, a combination of the Kabat definition and the Chothia definition, the AbM definition, or the contact definition of HVR.