Monoclonal Antibodies to Programmed Death 1 (PD-1)

Abstract

The present invention provides isolated monoclonal antibodies, particularly human monoclonal antibodies, that specifically bind to PD-1 with high affinity. Nucleic acid molecules encoding the antibodies of the invention, expression vectors, host cells and methods for expressing the antibodies of the invention are also provided. Immunoconjugates, bispecific molecules and pharmaceutical compositions comprising the antibodies of the invention are also provided. The invention also provides methods for detecting PD-1, as well as methods for treating various diseases, including cancer and infectious diseases, using anti-PD-1 antibodies. The present invention further provides methods for using a combination immunotherapy, such as the combination of anti-CTLA-4 and anti-PD-1 antibodies, to treat hyperproliferative disease, such as cancer. The invention also provides methods for altering adverse events related to treatment with such antibodies individually.

1 - 21 . (canceled) 
     
     
         22 . An isolated monoclonal antibody, or antigen-binding portion thereof, comprising a heavy chain variable region that comprises CDR1, CDR2, and CDR3 sequences; and a light chain variable region that comprises CDR1, CDR2, and CDR3 sequences, wherein:
 a) the heavy chain variable region CDR3 sequence comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 29, 30, 31, 32, 33, 34 and 35, and conservative modifications thereof,   b) the light chain variable region CDR3 sequence comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 50, 51, 52, 53, 54, 55 and 56, and conservative modifications thereof, and   c) the antibody or the antigen-binding portion thereof specifically binds to PD-1.   
     
     
         23 . The antibody of  claim 22 , wherein the heavy chain variable region CDR2 sequence comprises an amino acid sequence selected from the group consisting of amino acid sequences of SEQ ID NOs: 22, 23, 24, 25, 26, 27 and 28, and conservative modifications thereof, and the light chain variable region CDR2 sequence comprises an amino acid sequence selected from the group consisting of amino acid sequences of SEQ ID NOs: 43, 44, 45, 46, 47, 48 and 49, and conservative modifications thereof. 
     
     
         24 . The antibody of  claim 23 , wherein the heavy chain variable region CDR1 sequence comprises an amino acid sequence selected from the group consisting of amino acid sequences of SEQ ID NOs: 15, 16, 17, 18, 19, 20 and 21, and conservative modifications thereof; and the light chain variable region CDR1 sequence comprises an amino acid sequence selected from the group consisting of amino acid sequences of SEQ ID NOs: 36, 37, 38, 39, 40, 41 and 42, and conservative modifications thereof. 
     
     
         25 - 51 . (canceled) 
     
     
         52 . An isolated nucleic acid molecule encoding the antibody, or antigen-binding portion thereof, of  claim 22 . 
     
     
         53 . An expression vector comprising the nucleic acid molecule of  claim 52 . 
     
     
         54 . A host cell comprising the expression vector of  claim 53 . 
     
     
         55 . A transgenic mouse comprising immunoglobulin heavy and light chain transgenes, wherein the mouse expresses the antibody or the antigen-binding portion thereof of  claim 22 . 
     
     
         56 . A hybridoma prepared from the mouse of  claim 55 , wherein the hybridoma produces said antibody. 
     
     
         57 . A method of modulating an immune response in a subject comprising administering to the subject the antibody, or antigen-binding portion thereof, of  claim 22  such that the immune response in the subject is modulated. 
     
     
         58 - 63 . (canceled) 
     
     
         64 . A method of inhibiting growth of tumor cells in a subject, comprising administering to a subject the antibody, or antigen-binding portion thereof, of  claim 22  in an amount effective to inhibit growth of the tumor cells. 
     
     
         65 . A method of treating an infectious disease in a subject comprising administering to the subject the antibody, or antigen-binding portion thereof, of  claim 22  such that the subject is treated for the infectious disease. 
     
     
         66 - 68 . (canceled) 
     
     
         69 . A method for treating a hyperproliferative disease, which method comprises:
 (a) administering an anti-PD-1 antibody to a subject, and   (b) administering an anti-CTLA-4 antibody to the subject.   
     
     
         70 - 104 . (canceled) 
     
     
         105 . A method for preparing an anti-PD-1 antibody comprising:
 (a) providing: (i) a heavy chain variable region antibody sequence comprising a CDR1 sequence that is selected from the group consisting of SEQ ID NOs: 15, 16, 17, 18, 19, 20 and 21, a CDR2 sequence that is selected from the group consisting of SEQ ID NOs: 22, 23, 24, 25, 26, 27 and 28; and a CDR3 sequence that is selected from the group consisting of SEQ ID NOs: 29, 30, 31, 32, 33, 34 and 35; or (ii) a light chain variable region antibody sequence comprising a CDR1 sequence that is selected from the group consisting of SEQ ID NOs: 36, 37, 38, 39, 40, 41 and 42, a CDR2 sequence that is selected from the group consisting of SEQ ID NOs: 43, 44, 45, 46, 47, 48 and 49, and a CDR3 sequence that is selected from the group consisting of SEQ ID NOs: 50, 51, 52, 53, 54, 55 and 56;   (b) altering at least one amino acid residue within at least one variable region antibody sequence, said sequence being selected from the heavy chain variable region antibody sequence and the light chain variable region antibody sequence, to create at least one altered antibody sequence; and   (c) expressing the altered antibody sequence as a protein.   
     
     
         106 . The antibody of  claim 22 , wherein the antibody or antigen-binding portion thereof exhibits at least one of the following properties:
 (i) binds to human PD-1 with a K D  of 1×10 −7  M or less, wherein the K D  is measured by surface plasmon resonance (Biacore) analysis;   (ii) binds to human PD-1 with an on rate (k on ) of about 0.76×10 5  l/Ms or more, wherein the on rate (k on ) is measured by surface plasmon resonance (Biacore) analysis; and   (iii) binds to human PD-1 with an off rate (k off ) of about 4.5×10 −4  l/s or less, wherein the off rate (k off ) is measured by surface plasmon resonance (Biacore) analysis.   
     
     
         107 . The antibody of  claim 22 , wherein:
 (a) the heavy chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 15; the heavy chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 22; the heavy chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 29; the light chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 36; the light chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 43; and the light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 50;   (b) the heavy chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 16; the heavy chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 23; the heavy chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 30; the light chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 37; the light chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 44; and the light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 51;   (c) the heavy chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 17; the heavy chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 24; the heavy chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 31; the light chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 38; the light chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 45; and the light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 52;   (d) the heavy chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 18; the heavy chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 25; the heavy chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 32; the light chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 39; the light chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 46; and the light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 53;   (e) the heavy chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 19; the heavy chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 26; the heavy chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 33; the light chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 40; the light chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 47; and the light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 54;   (f) the heavy chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 20; the heavy chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 27; the heavy chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 34; the light chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 41; the light chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 48; and the light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 55; or   (g) the heavy chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 21; the heavy chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 28; the heavy chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 35 the light chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 42; the light chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 49; and the light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 56.   
     
     
         108 . The antibody of  claim 22 , wherein:
 (a) the heavy chain variable region comprises amino acids having the sequence set forth in SEQ ID NO: 1; and the light chain variable region comprises amino acids having the sequence set forth in SEQ ID NO: 8;   (b) the heavy chain variable region comprises amino acids having the sequence set forth in SEQ ID NO: 2; and the light chain variable region comprises amino acids having the sequence set forth in SEQ ID NO: 9;   (c) the heavy chain variable region comprises amino acids having the sequence set forth in SEQ ID NO: 3; and the light chain variable region comprises amino acids having the sequence set forth in SEQ ID NO: 10;   (d) the heavy chain variable region comprises amino acids having the sequence set forth in SEQ ID NO: 4; and the light chain variable region comprises amino acids having the sequence set forth in SEQ ID NO: 11;   (e) the heavy chain variable region comprises amino acids having the sequence set forth in SEQ ID NO: 5; and the light chain variable region comprises amino acids having the sequence set forth in SEQ DD NO: 12;   (f) the heavy chain variable region comprises amino acids having the sequence set forth in SEQ ID NO: 6; and the light chain variable region comprises amino acids having the sequence set forth in SEQ ID NO: 13; or   (g) the heavy chain variable region comprises amino acids having the sequence set forth in SEQ ID NO: 7; and the light chain variable region comprises amino acids having the sequence set forth in SEQ ID NO: 14.   
     
     
         109 . The antibody of  claim 22 , wherein the antibody or antigen-binding portion thereof is a humanized or human monoclonal antibody or an antigen-binding portion thereof. 
     
     
         110 . The antibody of  claim 22 , wherein the antibody further comprises a heavy chain constant region which is of a human IgG4. 
     
     
         111 . The antibody of  claim 22 , wherein the antibody or antigen-binding portion thereof comprises a light chain constant region which is a human kappa or lambda constant region. 
     
     
         112 . The antibody of  claim 22 , wherein the antigen-binding portion is a Fab, Fab′, F(ab′)2, Fv, or a single chain Fv fragment.