Antibodies specifically binding PD-1, TIM-3 or PD-1 and TIM-3 and their uses

1. An isolated antagonistic antibody or an antigen-binding portion thereof specifically binding PD-1, comprising a heavy chain complementarity determining region (HCDR) 1 a HCDR2 and a HCDR3 of SEQ ID NOs: 10, 14 and 17, respectively, and a light chain complementarity determining region (LCDR)-1, a LCDR2 and a LCDR3 of SEQ ID NOs: 23, 26 and 32, respectively.

2. The antibody or the antigen-binding portion thereof of claim 1 , comprising a heavy chain variable region (VH) of SEQ ID NO: 48 and a light chain variable region (VL) of SEQ ID NO: 56.

3. The antibody or the antigen-binding portion thereof of claim 1 , wherein the antibody is human or humanized.

4. The antibody or the antigen-binding portion thereof of claim 3 , wherein the antibody is

a) an IgG1 isotype, optionally comprising one, two, three, four, five, six, seven, eight, nine or ten substitutions in an Fc region;

b) an IgG2 isotype, optionally comprising one, two, three, four, five, six, seven, eight, nine or ten substitutions in the Fc region;

c) an IgG3 isotype, optionally comprising one, two, three, four, five, six, seven, eight, nine or ten substitutions in the Fc region;

d) an IgG4 isotype, optionally comprising one, two, three, four, five, six, seven, eight, nine or ten substitutions in the Fc region;

e) an IgG1 isotype comprising L234A, L235A, G237A, P238S, H268A, A330S and P331 S substitutions;

f) an IgG2 isotype comprising V234A, G237A, P238S, H268A, V309L, A330S and P331 S substitutions;

g) an IgG4 isotype comprising F234A, L235A, G237A, P238S and Q268A substitutions;

h) an IgG1 isotype comprising L234A, L235A or L234A and L235A substitutions;

i) an IgG4 isotype comprising F234A, L235A or F234A and L235A substitutions;

j) an IgG2 isotype comprising a V234A substitution;

k) an IgG4 isotype comprising a S228P substitution; or

l) an IgG4 isotype comprising S228P, F234A and L235A substitutions, wherein residue numbering is according to the EU Index.

5. The antibody or the antigen-binding portion thereof of claim 2 , comprising a heavy chain (HC) of SEQ ID NO: 72 and a light chain (LC) of SEQ ID NO: 73.

6. The antibody or the antigen-binding portion thereof of claim 2 , wherein the antibody is a bispecific antibody, optionally binding PD-Ll (SEQ ID NO: 5), PD-L2 (SEQ ID NO: 8), LAG-3 (SEQ ID NO: 293), TIM-3 (SEQ ID NO: 138), CEACAM-1 (SEQ ID NO: 296), CEACAM-5 (SEQ ID NO: 307), OX-40 (SEQ ID NO: 279), GITR (SEQ ID NO: 271), CD27 (SEQ ID NO: 280), VISTA (SEQ ID NO: 286), CD137 (SEQ ID NO: 281), TIGIT (SEQ ID NO:

301) or CTLA-4 (SEQ ID NO: 292).

7. A pharmaceutical composition comprising the antibody or the antigen-binding portion thereof of claim 2 and a pharmaceutically accepted carrier.

8. An isolated antagonistic antibody specifically binding PD-1 or an antigen-binding portion thereof, comprising

a) the HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 66, 67, 68, 69, 70 and 71, respectively;

b) the VH of SEQ ID NO: 64 and the VL of SEQ ID NO: 65; and/or

c) the HC of SEQ ID NO: 74 and the LC of SEQ ID NO: 75.

9. A pharmaceutical composition comprising the antibody or the antigen-binding portion thereof of claim 8 and a pharmaceutically accepted carrier.

10. A kit comprising the antibody of claim 2 .

11. The antibody or the antigen-binding portion thereof of claim 6 , wherein the bispecific antibody comprises a first domain specifically binding PD-1 and a second domain specifically binding TIM-3.

12. The antibody or the antigen-binding portion thereof of claim 11 , wherein

a) the first domain comprises the HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 10, 14, 17, 23, 26 and 32, respectively or SEQ ID NOs: 66, 67, 68, 69, 70 and 71, respectively; and

b) the second domain comprises the HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 97, 105, 115, 124, 133 and 143, respectively; or SEQ ID NOs: 91, 99, 108, 118, 127 and 136, respectively.

13. The antibody or the antigen-binding portion thereof of claim 12 , wherein the bispecific antibody binds TIM-3 within TIM-3 residues 32-47 (WGKGACPVFECGNVVL) (SEQ ID NO: 261), optionally further binding within TIM-3 residues 50-56 (DERDVNY) (SEQ ID NO: 262).

14. The antibody or the antigen-binding portion thereof of claim 13 , wherein

a. the first domain comprises the HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 10, 14, 17, 23, 26 and 32, respectively, and the second domain comprises the HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 91, 99, 108, 118, 127 and 136, respectively;

b. the first domain comprises the VH of SEQ ID NO: 48 and the VL of SEQ ID NO: 56, and the second domain comprises the VH of SEQ ID NO: 146 and the VL of SEQ ID NO: 156;

c. the antibody comprises a first heavy chain (HCl), a first light chain (LC1), a second heavy chain (HC2) and a second light chain (LC2) of SEQ ID NOs: 186, 188, 191 and 194, respectively;

d. the antibody comprises the HCl, the LC1, the HC2 and the LC2 of SEQ ID NOs: 186, 188, 248 and 194, respectively;

e. the antibody comprises the HCl, the LC1, the HC2 and the LC2 of SEQ ID NOs: 241, 188, 245 and 194, respectively; and/or

f. the antibody comprises the HCl, the LC1, the HC2 and the LC2 of SEQ ID NOs: 243, 188, 246 and 194, respectively.

15. The antibody or the antigen-binding portion thereof of claim 12 , wherein the bispecific antibody binds TIM-3 within TIM-3 residues 90-102 (RIQIPGIMNDEKF) (SEQ ID NO: 263), optionally further binding within TIM-3 residues 50-56 (DERDVNY) (SEQ ID NO: 262).

16. The antibody or the antigen-binding portion thereof of claim 15 , wherein

a. the first domain comprises the HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 10, 14, 17, 23, 26 and 32, respectively, and the second domain comprises the HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 97, 105, 115, 124, 133 and 143, respectively;

b. the first domain comprises the VH of SEQ ID NO: 48 and the VL of SEQ ID NO: 56, and the second domain comprises the VH of SEQ ID NO: 172 and the VL of SEQ ID NO: 173;

c. the antibody comprises a HCl, a LC1, a HC2 and a LC2 of SEQ ID NOs: 186, 188, 192 and 195, respectively;

d. the antibody comprises the HCl, the LC1, the HC2 and the LC2 of SEQ ID NOs: 241, 188, 244 and 195, respectively; and/or

e. the antibody comprises the HCl, the LC1, the HC2 and the LC2 of SEQ ID NOs: 243, 188, 247 and 195, respectively.

17. The antibody or the antigen-binding portion thereof of claim 11 , wherein the bispecific antibody is

a. an IgG1 isotype, optionally comprising one, two, three, four, five, six, seven, eight, nine or ten substitutions in an Fc region;

b. an IgG2 isotype, optionally comprising one, two, three, four, five, six, seven, eight, nine or ten substitutions in an Fc region;

c. an IgG3 isotype, optionally comprising one, two, three, four, five, six, seven, eight, nine or ten substitutions in an Fc region;

d. an IgG4 isotype, optionally comprising one, two, three, four, five, six, seven, eight, nine or ten substitutions in an Fc region;

e. an IgG1 isotype comprising L234A, L235A, G237A, P238S, H268A, A330S and P331 S substitutions;

f. an IgG2 isotype comprising V234A, G237A, P238S, H268A, V309L, A330S and P331 S substitutions;

g. an IgG4 isotype comprising F234A, L235A, G237A, P238S and Q268A substitutions;

h. an IgG1 isotype comprising L234A, L235A or L234A and L235A substitutions;

i. an IgG4 isotype comprising F234A, L235A or F234A and L235A substitutions;

j. an IgG2 isotype comprising a V234A substitution;

k. an IgG4 isotype comprising a S228P substitution;

l. an IgG4 isotype comprising S228P, F234A and L235A substitutions;

m. an IgG2 isotype comprising F405L and R409K substitutions; or

n. an IgG1 isotype comprising F405L and K409R substitutions, wherein residue numbering is according to the EU Index.

18. A pharmaceutical composition comprising the bispecific antibody of claim 12 and a pharmaceutically accepted carrier.

19. An isolated antagonistic antibody specifically binding PD-1 or an antigen-binding portion thereof, comprising a HCDR1, a HCDR2 and a HCDR3 of SEQ ID NOs:

11, 15 and 18, respectively and a LCDR1, a LCDR2 and a LCDR3 of SEQ ID NOs: 20, 30 and 32, respectively.

20. The antibody or the antigen-binding portion thereof of claim 19 , comprising a VH and a VL of SEQ ID NOs: 45 and 60, respectively.

21. The antibody or the antigen-binding portion thereof of claim 20 , wherein the antibody is an IgG4 isotype comprising a S228P substitution.