Anti-PD-1 antibodies and compositions

1. Isolated nucleic acid molecule(s) comprising a nucleotide sequence that encodes the heavy chain sequence, and a nucleotide sequence that encodes the light chain sequence, of an anti-PD-1 antibody or an antigen-binding portion thereof that comprises the H-CDR1-3 and L-CDR1-3 amino acid sequences of:

a) SEQ ID NOs: 18, 19, 20, 21, 22, and 23, respectively;

b) SEQ ID NOs: 24, 25, 26, 27, 28, and 29, respectively;

c) SEQ ID NOs: 30, 31, 32, 33, 34, and 35, respectively;

d) SEQ ID NOs: 36, 37, 38, 39, 40, and 41, respectively;

e) SEQ ID NOs: 42, 43, 44, 45, 46, and 47, respectively;

f) SEQ ID NOs: 48, 49, 50, 51, 52, and 53, respectively;

g) SEQ ID NOs: 54, 55, 56, 57, 58, and 59, respectively; or

h) SEQ ID NOs: 60, 61, 62, 63, 64, and 65, respectively.

2. Vector(s) comprising the isolated nucleic acid molecule(s) of claim 1 , wherein said vector(s) further comprise an expression control sequence.

3. A host cell comprising the isolated nucleic acid molecule(s) of claim 1 .

4. A method for producing an anti-PD-1 antibody or an antigen-binding portion thereof, comprising providing a host cell of claim 3 , cultivating said host cell under conditions suitable for expression of the antibody or portion, and isolating the resulting antibody or portion.

5. The isolated nucleic acid molecule(s) of claim 1 , wherein said anti-PD-1 antibody comprises a heavy chain variable domain and a light chain variable domain with amino acid sequences at least 90% identical to the amino acid sequences of:

a) SEQ ID NOs: 2 and 3, respectively;

b) SEQ ID NOs: 4 and 5, respectively;

c) SEQ ID NOs: 4 and 66, respectively;

d) SEQ ID NOs: 6 and 7, respectively;

e) SEQ ID NOs: 8 and 9, respectively;

f) SEQ ID NOs: 10 and 11, respectively;

g) SEQ ID NOs: 12 and 13, respectively;

h) SEQ ID NOs: 14 and 15, respectively; or

i) SEQ ID NOs: 16 and 17, respectively.

6. The isolated nucleic acid molecule(s) of claim 1 , wherein said anti-PD-1 antibody comprises a heavy chain variable domain and a light chain variable domain having the amino acid sequences of:

a) SEQ ID NOs: 2 and 3, respectively;

b) SEQ ID NOs: 4 and 5, respectively;

c) SEQ ID NOs: 4 and 66, respectively;

d) SEQ ID NOs: 6 and 7, respectively;

e) SEQ ID NOs: 8 and 9, respectively;

f) SEQ ID NOs: 10 and 11, respectively;

g) SEQ ID NOs: 12 and 13, respectively;

h) SEQ ID NOs: 14 and 15, respectively; or

i) SEQ ID NOs: 16 and 17, respectively.

7. The isolated nucleic acid molecule(s) of claim 1 , wherein said anti-PD-1 antibody is of isotype IgG.

8. The isolated nucleic acid molecule(s) of claim 7 , wherein:

a) the anti-PD-1 antibody is of isotype subtype IgG1 and one or both of the amino acid residues at positions 234 and 235 are mutated to Ala; or

b) the anti-PD-1 antibody is of isotype subtype IgG4 and the amino acid residue at position 228 is mutated to Pro.

9. The isolated nucleic acid molecule(s) of claim 1 , wherein the anti-PD-1 antibody or portion has at least one of the following properties:

a) binds to human PD-1 with a K D of 750 pM or less;

b) binds to cynomolgus PD-1 with a K D of 7 nM or less;

c) binds to mouse PD-1 with a K D of 1 nM or less;

d) does not bind to rat PD-1;

e) increases IL-2 secretion in an SEB whole blood assay;

f) increases IFN-γ secretion in a one-way mixed lymphocyte reaction assay;

g) inhibits the interaction of PD-1 with PD-L1 by at least 60% at a concentration of 10 μg/mL in a flow cytometric competition assay;

h) blocks binding of PD-L1 and PD-L2 to PD-1 by at least 90% at a concentration of 10 μg/mL as determined by Bio-Layer Interferometry analysis; and

i) inhibits tumor growth in vivo.

10. The isolated nucleic acid molecule(s) of claim 1 , wherein the anti-PD-1 antibody comprises:

a) a heavy chain (HC) comprising the amino acid sequences of SEQ ID NOs: 2 and 67 and a light chain (LC) comprising the amino acid sequences of SEQ ID NOs: 3 and 68;

b) an HC comprising the amino acid sequences of SEQ ID NOs: 4 and 67 and an LC comprising the amino acid sequences of SEQ ID NOs: 5 and 68;

c) an HC comprising the amino acid sequences of SEQ ID NOs: 4 and 67 and an LC comprising the amino acid sequences of SEQ ID NOs: 66 and 68;

d) an HC comprising the amino acid sequences of SEQ ID NOs: 6 and 67 and an LC comprising the amino acid sequences of SEQ ID NOs: 7 and 68;

e) an HC comprising the amino acid sequences of SEQ ID NOs: 8 and 67 and an LC comprising the amino acid sequences of SEQ ID NOs: 9 and 68;

f) an HC comprising the amino acid sequences of SEQ ID NOs: 10 and 67 and an LC comprising the amino acid sequences of SEQ ID NOs: 11 and 68;

g) an HC comprising the amino acid sequences of SEQ ID NOs: 12 and 67 and an LC comprising the amino acid sequences of SEQ ID NOs: 13 and 68;

h) an HC comprising the amino acid sequences of SEQ ID NOs: 14 and 67 and an LC comprising the amino acid sequences of SEQ ID NOs: 15 and 68; or

i) an HC comprising the amino acid sequences of SEQ ID NOs: 16 and 67 and an LC comprising the amino acid sequences of SEQ ID NOs: 17 and 68.

11. Isolated nucleic acid molecule(s) comprising a nucleotide sequence that encodes the heavy chain sequence, and a nucleotide sequence that encodes the light chain sequence, of an anti-PD-1 antibody or an antigen-binding portion thereof that comprises the H-CDR1-3 and L-CDR1-3 amino acid sequences of SEQ ID NOs: 18, 19, 20, 21, 22, and 23, respectively.

12. Vector(s) comprising the isolated nucleic acid molecule(s) of claim 11 , wherein said vector(s) further comprise an expression control sequence.

13. A host cell comprising the isolated nucleic acid molecule(s) of claim 11 .

14. A method for producing an anti-PD-1 antibody or an antigen-binding portion thereof, comprising providing a host cell of claim 13 , cultivating said host cell under conditions suitable for expression of the antibody or portion, and isolating the resulting antibody or portion.

15. Isolated nucleic acid molecule(s) comprising a nucleotide sequence that encodes the heavy chain sequence, and a nucleotide sequence that encodes the light chain sequence, of an anti-PD-1 antibody that comprises a heavy chain (HC) comprising the amino acid sequences of SEQ ID NOs: 2 and 67 and a light chain (LC) comprising the amino acid sequences of SEQ ID NOs: 3 and 68.

16. Vector(s) comprising the isolated nucleic acid molecule(s) of claim 15 , wherein said vector(s) further comprise an expression control sequence.

17. A host cell comprising the isolated nucleic acid molecule(s) of claim 15 .

18. A method for producing an anti-PD-1 antibody, comprising providing a host cell of claim 17 , cultivating said host cell under conditions suitable for expression of the antibody, and isolating the resulting antibody.