Novel Monoclonal Antibodies to Programmed DEATH (PD-1)

Abstract

The present invention provides PD-1 monoclonal antibodies, particularly human monoclonal antibodies of PD-1, which specifically bind to PD-1 with high affinity and comprise a heavy chain and a light chain. The present invention further provides nucleic acid sequence encoding the antibodies of the invention, cloning or expression vectors, host cells and methods for expressing or isolating the antibodies. Immunoconjugates, therapeutic compositions comprising the antibodies of the invention are also provided. The invention also provides methods for treating various cancers with anti-PD-1 antibodies.

1 . An antibody or antigen binding fragment thereof that binds to an epitope of PD-1 comprising amino acids at positions 128, 129, 130, 131 and 132 and at least one of amino acids at positions 35, 64, 82, 83 of SEQ ID NO: 24. 
     
     
         2 . (canceled) 
     
     
         3 . The antibody or the antigen binding fragment thereof of  claim 1 , wherein the murine PD-1 is mouse or rat PD-1. 
     
     
         4 . The antibody or the antigen binding fragment thereof of  claim 1 , wherein the antibody
 a) binds to human PD-1 with a K D  of 2.15E-10 M or less; and   b) binds to mouse PD-1 with a K D  of 1.67E-08 M or less.   
     
     
         5 . The antibody of any of  claim 1 , wherein the antibody exhibits at least one of the following properties:
 a) binds to human PD-1 with a K D  of between 4.32E-10 M and 2.15E-10 M and to mouse PD-1 with a K D  of between 5.39E-08 M and 1.67E-08 M;   b) does not substantially bind to human CD28, CTLA-4;   c) increases T-cell proliferation;   d) increases interferon-gamma production; or   e) increases interleukin-2 secretion.   
     
     
         6 . An antibody or an antigen binding fragment thereof, comprising an amino acid sequence that is at least 70%, 80%, 90% or 95% homologous to a sequence selected from a group consisting of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8 and 9, wherein the antibody specifically binds to PD-1. 
     
     
         7 . (canceled) 
     
     
         8 . The antibody, or an antigen-binding fragment thereof  claim 6 , comprising:
 a) a variable region of a heavy chain having an amino acid sequence that is at least 70%, 80%, 90% or 95% homologous to a sequence selected from a group consisting of SEQ ID NOs: 1 and 2; and   b) a variable region of a light chain having an amino acid sequence that is at least 70%, 80%, 90% or 95% homologous to a sequence selected from a group consisting of SEQ ID NOs: 3, 4, 5, 6, 7, 8 and 9,   wherein the antibody specifically binds to PD-1.   
     
     
         9 . The antibody or an antigen binding fragment thereof of  claim 8 , comprising:
 a) a variable region of a heavy chain having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 and 2; and   b) a variable region of a light chain having an amino acid sequence selected from the group consisting of SEQ ID NOs: 3, 4, 5, 6, 7, 8 and 9,   wherein the antibody specifically binds to PD-1.   
     
     
         10 . The antibody or an antigen binding fragment thereof of  claim 6 , comprising a complementarity-determining region (CDR) having an amino acid sequence selected from the group consisting of SEQ ID NOs: 10-23,
 wherein the antibody specifically binds to PD-1.   
     
     
         11 . The antibody, or antigen-binding fragment thereof gf  claim 10 , comprising:
 a heavy chain variable region comprising CDR1, CDR2, and CDR3 sequences; and   a light chain variable region comprising CDR1, CDR2, and CDR3 sequences,   wherein the heavy chain variable region CDR3 sequence comprises a sequence selected from a group consisting of SEQ ID NOs: 12 and 13, and conservative modifications thereof,   wherein the antibody specifically binds to PD-1.   
     
     
         12 . The antibody of  claim 10 , wherein the light chain variable region CDR3 sequence comprises an amino acid sequence selected from a group consisting of SEQ ID NOs: 20, 21, 22 and 23, and conservative modifications thereof. 
     
     
         13 . The antibody of  claim 10 , wherein the heavy chain variable region CDR2 sequence comprises an amino acid sequence selected from a group consisting of amino acid sequences of SEQ ID NO: 11, and conservative modifications thereof. 
     
     
         14 . The antibody of  claim 10 , wherein the light chain variable region CDR2 sequence comprises an amino acid sequence selected from a group consisting of amino acid sequences of SEQ ID NO: 19, and conservative modifications thereof. 
     
     
         15 . The antibody of  claim 10 , wherein the heavy chain variable region CDR1 sequence comprises an amino acid sequence selected from a group consisting of amino acid sequences of SEQ ID NO: 10, and conservative modifications thereof. 
     
     
         16 . The antibody of  claim 10 , wherein the light chain variable region CDR1 sequence comprises an amino acid sequence selected from a group consisting of amino acid sequences of SEQ ID NOs: 14, 15, 16, 17 and 18, and conservative modifications thereof. 
     
     
         17 . The antibody of  claim 6 , wherein the antibody is a chimeric or humanized or human antibody. 
     
     
         18 . The antibody of  claim 6 , wherein the antibody exhibits at least one of the following properties:
 a) binds to human PD-1 with a K D  of 2.15E-10 M or less and to mouse PD-1 with a K D  of 1.67E-08 M or less;   b) does not substantially bind to human CD28, CTLA-4;   c) increases T-cell proliferation;   d) increases interferon-gamma production; or   e) increases interleukin-2 secretion.   
     
     
         19 . A nucleic acid molecule encoding the antibody, or the antigen binding fragment of  claim 6 . 
     
     
         20 . A cloning or expression vector comprising the nucleic acid molecule of  claim 19 . 
     
     
         21 . A host cell comprising one or more cloning or expression vectors of  claim 20 . 
     
     
         22 . A process for the production of the antibody of  claim 6 , comprising culturing the host cell of  claim 21  and isolating the antibody. 
     
     
         23 . The process of  claim 22 , wherein the antibody is prepared through immunization in SD rat with human PD-1 extracellular domain and mouse PD-1 extracellular domain. 
     
     
         24 . A transgenic mouse comprising human immunoglobulin heavy and light chain transgenes, wherein the mouse expresses the antibody of  claim 6 . 
     
     
         25 . A hybridoma prepared from the mouse of  claim 24 , wherein the hybridoma produces said antibody. 
     
     
         26 . A pharmaceutical composition comprising the antibody, or the antigen binding fragment of  claim 6 , and one or more of a pharmaceutically acceptable excipient, diluent or carrier. 
     
     
         27 . An immunoconjugate comprising the antibody, or antigen-binding fragment thereof, according to  claim 6 , linked to a therapeutic agent. 
     
     
         28 . A pharmaceutical composition comprising the immunoconjugate of  claim 27  and a pharmaceutically acceptable excipient, diluent or carrier. 
     
     
         29 . A method for preparing an anti-PD-1 antibody or an antigen-binding fragment thereof comprising:
 a) providing:   (i) a heavy chain variable region antibody sequence comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 and 2;   ; and/or   (ii) a light chain variable region antibody sequence comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 3, 4, 5, 6, 7, 8 and 9,   ; and   b) expressing the antibody sequence as a protein.   
     
     
         30 . A method of modulating an immune response in a subject comprising administering to the subject the antibody, or antigen binding fragment of  claim 6 . 
     
     
         31 . (canceled) 
     
     
         32 . A method of inhibiting growth of tumor cells in a subject, comprising administering to the subject a therapeutically effective amount of the antibody, or the antigen-binding fragment of  claim 6  to inhibit growth of the tumor cells. 
     
     
         33 . The method of  claim 32 , wherein the tumor cells are of a cancer selected from a group consisting of melanoma, renal cancer, prostate cancer, breast cancer, colon cancer, lung cancer, bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, and rectal cancer. 
     
     
         34 . The method of  claim 32 , wherein the antibody is a chimeric antibody or humanized antibody.