ANTIBODIES TO HUMAN PROGRAMMED DEATH RECEPTOR PD-1

• US 20130108651A1
• Filed: 12/19/2012
• Published: 05/02/2013
• Est. Priority Date: 06/18/2007
• Status: Active Grant

• Current Assignee: Merck Sharp & Dohme BV (Merck & Co., Inc.)
• Original Assignee: MSD Oss BV (Merck & Co., Inc.)
• Inventors: Carven, Gregory John, Dulos, Gradus Johannes, Van Eenenneem, Hans
• Granted Patent: US 8,900,587 B2
• Time in Patent Office: Days
• US Class Current: 424/172.1
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: C07K 16/2803 against the immunoglobulin superfamily
• CPC Class Codes: C07K 16/2818 against CD28 or CD152
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: G01N 2333/55 IL-2
• CPC Class Codes: G01N 2800/52 Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
• CPC Class Codes: G01N 33/57488 involving compounds identifable in body fluids
• CPC Class Codes: G01N 33/6869 Interleukin

Abstract

Antibodies which block binding of hPD-1 to hPD-L1 or hPD-L2 and their variable region sequences are disclosed. A method of increasing the activity (or reducing downmodulation) of an immune cell through the PD-1 pathway is also disclosed.

Claims

1. An isolated antibody or antibody fragment which binds to human PD-1 comprising:

a. at least one CDR selected from the group consisting of SEQ ID NOs;

9, 10, 11, 15, 16 and 17, or a variant of any said sequence; and

/or

b. at least one CDR selected from the group consisting of SEQ ID NOs;

12, 13, 14, 18, 19 and 20, or a variant of any said sequence.

2. The antibody or antibody fragment of claim 1, comprising:

a. light chain CDRs SEQ ID NOs;

9, 10 and 11, or variants of any said sequences; and

heavy chain CDRs SEQ ID NOs;

12, 13 and 14, or variants of any said sequences;

or

b. light chain CDRs SEQ ID NOs;

15, 16 and 17 or variants of any said sequences; and

heavy chain CDRs SEQ ID NOs;

18, 19 and 20 or variants of any said sequences.

3. The antibody or antibody fragment of claim 2, comprising:

a. a heavy chain variable region comprising an amino acid sequence selected from the group consisting of;

i. SEQ ID NO;

5 or a variant thereof;

ii. SEQ ID NO;

7 or a variant thereof;

iii. amino acid residues 20 to 139 of SEQ ID NO;

30 or a variant thereof; and

iv. an amino acid sequence having at least 90% homology to amino acid residues 20 to 139 of SEQ ID NO;

30;

and further comprising

b. a light chain variable region comprising an amino acid sequence selected from the group consisting of;

i. SEQ ID NO;

6 or a variant thereof;

ii. SEQ ID NO;

8 or a variant thereof;

iii. amino acid residues 20 to 130 of SEQ ID NO;

32 or a variant thereof;

iv. amino acid residues 20 to 130 of SEQ ID NO;

33 or a variant thereof;

v. amino acid residues 20 to 130 of SEQ ID NO;

34 or a variant thereof; and

vi. an amino acid sequence having at least 90% homology to amino acid residues 20 to 130 of SEQ ID NO;

32, 33 or 34.

4. The antibody of claim 1, comprising:

a. a heavy chain comprising an amino acid sequence selected from the group consisting of;

i. amino acid residues 20 to 466 of SEQ ID NO;

31 or a variant thereof, and

ii. amino acid residues 20 to 469 of SEQ ID NO;

35 or a variant thereof;

and

b. a light chain comprising an amino acid sequence selected from the group consisting of;

i. amino acid residues 20 to 237 of SEQ ID NO;

36 or a variant thereof;

ii. amino acid residues 20 to 237 of SEQ ID NO;

37 or a variant thereof, and

iii. amino acid residues 20 to 237 of SEQ ID NO;

38 or a variant thereof.

5. The antibody or antibody fragment of claim 1, wherein any of said variant(s) may comprise up to three conservatively modified amino acid substitutions.

6. The antibody of claim 3, further comprising:

a. a human heavy chain constant region or a variant thereof, wherein the variant comprises up to 20 conservatively modified amino acid substitutions; and

/or

b. a human light chain constant region or a variant thereof, wherein the variant comprises up to 20 conservatively modified amino acid substitutions.

7. The antibody of claim 6, wherein the human heavy chain constant region comprises a γ

4 or γ

1 human heavy chain constant region or a variant thereof, wherein the variant comprises up to 20 conservatively modified amino acid substitutions.

8. The antibody or antibody fragment of claim 5, wherein the antibody or antibody fragment:

a. binds human PD-1 with a K_D of about 100 pM or lower;

b. binds human PD-1 with a K_D of about 30 pM or lower;

c. binds to human PD-1 with about the same K_D as an antibody having a heavy chain comprising the amino acid sequence of SEQ ID NO;

31 and a light chain comprising the amino acid sequence of SEQ ID NO;

32;

d. binds to human PD-1 with about the same K_D as an antibody having a heavy chain comprising the amino acid sequence of SEQ ID NO;

31 and a light chain comprising the amino acid sequence of SEQ ID NO;

33;

e. binds to human PD-1 with a k_assoc of about 7.5×

10⁵ 1/M·

s or faster;

f. binds to human PD-1 with a k_assoc of about 1×

10⁶ 1/M·

s or faster;

g. binds to human PD-1 with a k_assoc of about 2×

10<sup>−

5</sup> 1/s or slower;

h. binds to human PD-1 with a k_dissoc of about 2.7×

10<sup>−

5</sup> 1/s or slower;

i. binds to human PD-1 with a k_dissoc of about 3×

10<sup>−

5</sup> 1/s or slower;

or

j. blocks binding of human PD-L1 or human PD-L2 to human PD-1 with an IC₅₀ of about 1 nM or lower.

9. An isolated antibody or antibody fragment which competes for a binding epitope on PD-1 with antibody hPD-1.08A or antibody hPD-1.09A, and has a one of the following characteristics:

a. binds human PD-1 with a K_D of about 100 pM or lower;

b. binds human PD-1 with a K_D of about 30 pM or lower;

c. binds to human PD-1 with about the same K_D as an antibody having a heavy chain comprising the amino acid sequence of SEQ ID NO;

31 and a light chain comprising the amino acid sequence of SEQ ID NO;

32;

d. binds to human PD-1 with about the same K_D as an antibody having a heavy chain comprising the amino acid sequence of SEQ ID NO;

31 and a light chain comprising the amino acid sequence of SEQ ID NO;

33;

e. binds to human PD-1 with a k_assoc of about 7.5×

10⁵ 1/M·

s or faster;

f. binds to human PD-1 with a k_assoc of about 1×

10⁶ 1/M·

s or faster;

g. binds to human PD-1 with a k_assoc of about 2×

10<sup>−

5</sup> 1/s or slower;

h. binds to human PD-1 with a k_dissoc of about 2.7×

10<sup>−

5</sup> 1/s or slower;

i. binds to human PD-1 with a k_dissoc of about 3×

10<sup>−

5</sup> 1/s or slower;

or

j. blocks binding of human PD-L1 or human PD-L2 to human PD-1 with an IC₅₀ of about 1 nM or lower.

10. An isolated polynucleotide encoding an antibody or antibody fragment which binds to human PD-1, wherein the antibody or antibody fragment comprises

a. at least one CDR selected from the group consisting of SEQ ID NOs:

9, 10, 11, 15, 16 and 17; and

/or

b. at least one CDR selected from the group consisting of SEQ ID NOs;

12, 13, 14, 18, 19 and 20.

11. The isolated polynucleotide of claim 10, which comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs:

221, 22, 23, 24, 25, 26, 27, 28 and 29.

12. An expression vector comprising the isolated polynucleotide of claim 10.

13. A host cell comprising the expression vector of claim 12.

14. A method of producing an antibody or antibody fragment that binds to PD-1 comprising:

a. culturing the host cell of claim 13 in culture medium under conditions wherein the nucleic acid sequence is expressed, thereby producing polypeptides comprising the light and heavy chain variable regions; and

b. recovering the polypeptides from the host cell or culture medium.

15. A composition comprising an antibody or antibody fragment thereof in combination with a pharmaceutically acceptable carrier or diluent, wherein said antibody is h409A11, h409A16 or h409A17.

16. A method of increasing the activity of an immune cell, comprising administering to a subject in need thereof a therapeutically effective amount of an antibody or antibody fragment which binds to human PD-1, wherein the antibody or antibody fragment comprises

a. at least one CDR selected from the group consisting of SEQ ID NOs:

9, 10, 11, 15, 16 and 17; and

/or

b. at least one CDR selected from the group consisting of SEQ ID NOs;

12, 13, 14, 18, 19 and 20.

17. The method of claim 16, wherein said method is used for:

a. the treatment of cancer;

b. the treatment of an infection or infectious disease;

or

c. as a vaccine adjuvant.

18. The method of claim 17, further comprising measuring T cell activation ex vivo in a sample derived from the subject, wherein an increase in T cell activity indicates that the treatment should be continued or predicts the likelihood that the treatment will be successful.

19. The method of claim 18, wherein the increase in T cell activity is determined by:

a. measuring SEB induced production of one or more cytokines selected from the group consisting of;

IL-2, TNFα

, IL-17, IFNγ

, GM-CSF, RANTES, IL-6, IL-8, IL-5 and IL-13;

or

b. measuring TT induced production of a cytokine selected from the group consisting of;

IL-2, TNFα

, IL-17, IFNγ

, GM-CSF, RANTES, IL-6, IL-8, IL-5 and IL-13.