Glucocorticoid-induced tumor necrosis factor receptor (gitr) antibodies and methods of use thereof

Abstract

The present invention comprises human monoclonal antibodies that bind to GITR (also known as glucocorticoid-induced tumor necrosis factor receptor) Binding of the invented antibody to GITR inhibits binding of its ligand, GITR-L, and can be used to treat cancer.

What is claimed is: 
     
         1 . An isolated humanized monoclonal antibody or antigen-binding fragment thereof that binds to the human-glucocorticoid-induced tumor necrosis factor receptor (GITR) comprising:
 a. a variable heavy chain region comprising the amino acid sequence of SEQ ID NO: 2, and a variable light chain region comprising the amino acid sequence of SEQ ID NO: 4;   b. a variable heavy chain region comprising the amino acid sequence of SEQ ID NO: 6, and a variable light chain region comprising the amino acid sequence of SEQ ID NO: 8;   c. a variable heavy chain region comprising the amino acid sequence of SEQ ID NO: 10, and a variable light chain region comprising the amino acid sequence of SEQ ID NO: 12;   d. a variable heavy chain region comprising the amino acid sequence of SEQ ID NO: 14, and a variable light chain region comprising the amino acid sequence of SEQ ID NO: 16;   e. a variable heavy chain region comprising the amino acid sequence of SEQ ID NO: 18, and a variable light chain region comprising the amino acid sequence of SEQ ID NO: 20;   f. a variable heavy chain region comprising the amino acid sequence of SEQ ID NO: 22, and a variable light chain region comprising the amino acid sequence of SEQ ID NO: 24;   g. a variable heavy chain region comprising the amino acid sequence of SEQ ID NO: 26, and a variable light chain region comprising the amino acid sequence of SEQ ID NO: 28;   h. a variable heavy chain region comprising the amino acid sequence of SEQ ID NO: 30, and a variable light chain region comprising the amino acid sequence of SEQ ID NO: 32;   i. a variable heavy chain region comprising the amino acid sequence of SEQ ID NO: 34, and a variable light chain region comprising the amino acid sequence of SEQ ID NO: 36;   j. a variable heavy chain region comprising the amino acid sequence of SEQ ID NO: 38, and a variable light chain region comprising the amino acid sequence of SEQ ID NO: 40; or   k. a variable heavy chain region comprising the amino acid sequence of SEQ ID NO: 42, and a variable light chain region comprising the amino acid sequence of SEQ ID NO: 44.   
     
     
         2 . An isolated humanized monoclonal antibody or antigen-binding fragment thereof wherein the antibody or antigen-binding fragment comprises:
 (a) a variable heavy chain complementarity determining region 1, 2, or 3 (VH-CDR) comprising the amino acid sequences of SEQ ID NO. 45, 46 or 47, respectively; and, a variable light chain complementarity determining region 1, 2 or 3 (VL-CDR) comprising the amino acid sequences of SEQ ID NO. 48, 49, or 50;   (b) a variable heavy chain complementarity determining region 1, 2, or 3 (VH-CDR) comprising the amino acid sequences of SEQ ID NO. 51, 52, or 53, respectively; and, a variable light chain complementarity determining region 1, 2 or 3 (VL-CDR) comprising the amino acid sequences of SEQ ID NO. 54, 55, or 56;   (c) a variable heavy chain complementarity determining region 1, 2, or 3 (VH-CDR) comprising the amino acid sequences of SEQ ID NO. 57, 58, or 59, respectively; and, a variable light chain complementarity determining region 1, 2 or 3 (VL-CDR) comprising the amino acid sequences of SEQ ID NO. 60, 61, or 62;   (d) a variable heavy chain complementarity determining region 1, 2, or 3 (VH-CDR) comprising the amino acid sequences of SEQ ID NO. 63, 64, or 65, respectively; and, a variable light chain complementarity determining region 1, 2 or 3 (VL-CDR) comprising the amino acid sequences of SEQ ID NO. 66, 67, or 68;   (e) a variable heavy chain complementarity determining region 1, 2, or 3 (VH-CDR) comprising the amino acid sequences of SEQ ID NO. 69, 70, or 71, respectively; and, a variable light chain complementarity determining region 1, 2 or 3 (VL-CDR) comprising the amino acid sequences of SEQ ID NO. 72, 73, or 74;   (f) a variable heavy chain complementarity determining region 1, 2, or 3 (VH-CDR) comprising the amino acid sequences of SEQ ID NO. 75, 76, or 77, respectively; and, a variable light chain complementarity determining region 1, 2 or 3 (VL-CDR) comprising the amino acid sequences of SEQ ID NO. 78, 79, or 80;   (g) a variable heavy chain complementarity determining region 1, 2, or 3 (VH-CDR) comprising the amino acid sequences of SEQ ID NO. 81, 82, or 83, respectively; and, a variable light chain complementarity determining region 1, 2 or 3 (VL-CDR) comprising the amino acid sequences of SEQ ID NO. 84, 85, or 86;   (h) a variable heavy chain complementarity determining region 1, 2, or 3 (VH-CDR) comprising the amino acid sequences of SEQ ID NO. 87, 88, or 89, respectively; and, a variable light chain complementarity determining region 1, 2 or 3 (VL-CDR) comprising the amino acid sequences of SEQ ID NO. 90, 91, or 92;   (i) a variable heavy chain complementarity determining region 1, 2, or 3 (VH-CDR) comprising the amino acid sequences of SEQ ID NO. 93, 94, or 95, respectively; and, a variable light chain complementarity determining region 1, 2 or 3 (VL-CDR) comprising the amino acid sequences of SEQ ID NO. 96, 97, or 98;   (j) a variable heavy chain complementarity determining region 1, 2, or 3 (VH-CDR) comprising the amino acid sequences of SEQ ID NO. 99, 100, or 101, respectively; and, a variable light chain complementarity determining region 1, 2 or 3 (VL-CDR) comprising the amino acid sequences of SEQ ID NO. 102, 103, or 104; or   (k) a variable heavy chain complementarity determining region 1, 2, or 3 (VH-CDR) comprising the amino acid sequences of SEQ ID NO. 105, 106, or 107, respectively; and, a variable light chain complementarity determining region 1, 2 or 3 (VL-CDR) comprising the amino acid sequences of SEQ ID NO. 108, 109, or 110   wherein said antibody or antibody binding fragment binds human-glucocorticoid-induced tumor necrosis factor receptor (GITR)   
     
     
         3 . The antibody of  claim 1 , wherein said antibody is monovalent or bivalent. 
     
     
         4 . The antibody of  claim 1 , wherein said antibody is a single chain antibody. 
     
     
         5 . The antibody of  claim 1 , wherein said antibody has a binding affinity within the range of 10 −5  M to 10 −12  M. 
     
     
         6 . The antibody of  claim 1 , wherein said antibody has a IgG4 heavy chain constant region. 
     
     
         7 . The antibody of  claim 1 , wherein the Fc region contains mutations at amino acid positions 234 and 235. 
     
     
         8 . The antibody of  claim 7 , wherein the mutations are L234A and L235A. 
     
     
         9 . The antibody according to  claim 1  wherein said antibody is a bi-specific antibody that also binds to a tumor-associated antigen, a cytokine or a cell surface receptor. 
     
     
         10 . The antibody according to any one of preceding claims linked to a therapeutic agent. 
     
     
         11 . The antibody of  claim 10 , wherein said therapeutic agent is a toxin, a radiolabel, a siRNA, a small molecule, or a cytokine. 
     
     
         12 . A cell producing the antibody of any one of  claims 1 - 11 . 
     
     
         13 . A method of depleting regulatory T-cells in a subject, comprising administering to a subject in need thereof a composition comprising an antibody according to any one of  claims 1 - 11 . 
     
     
         14 . A method of augmenting an immune response to an antigen comprising administering to a subject in need thereof a composition comprising an antibody of any one of  claims 1 - 11 . 
     
     
         15 . The method of  claim 14 , wherein said antigen is a viral antigen, a bacterial antigen or a tumor associated antigen. 
     
     
         16 . The method of  claim 14 , wherein said administration of said antibody causes an increase in antigen specific T cell activity. 
     
     
         17 . The method of  claim 14 , wherein said administration of said antibody causes an increase NK cell cytoxicity. 
     
     
         18 . The method of  claim 14 , further comprising administering to said subject IL-15. 
     
     
         19 . A method of treating or alleviating a symptom of cancer, comprising administering to a subject in need thereof a composition comprising an antibody according to any one of  claims 1 - 11 . 
     
     
         20 . The method of  claim 19 , wherein said cancer is a cancer in which GITR or its ligand, GITR-L, is overexpressed. 
     
     
         21 . The method of  claim 20 , comprising further administering to said subject a cytokine or a chemotherapeutic agent. 
     
     
         22 . The method of  claim 21 , wherein the cytokine is IL-15. 
     
     
         23 . A nucleic acid comprising the nucleic acid sequence of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43. 
     
     
         24 . A nucleic acid encoding the polypeptide of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44. 
     
     
         25 . A polypeptide comprising the amino acid sequence of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44. 
     
     
         26 . A vector comprising the nucleic acid  claim 23  or  24 . 
     
     
         27 . A cell comprising the vector of  claim 26 .