Human antibodies to clostridium difficile toxins

Abstract

The present invention provides fully human antibodies that bind to either toxin A or toxin B of Clostridium difficile , or to both toxin A and toxin B, compositions comprising the antibodies and methods of use. The antibodies of the invention are useful for neutralizing the toxins from C. difficile , thus providing a means of treating the disease and symptoms associated with a C. difficile infection, including the treatment of diarrhea, or pseudomembranous colitis caused by C. difficile . The antibodies may also prevent the severity and/or duration of the primary disease, or may prevent the number, duration, and/or the severity of recurrences, or relapses of the disease attributed to the presence of C. difficile . The antibodies of the invention may also be useful for diagnosis of an infection by C. difficile.

What is claimed is: 
     
         1 . An isolated antibody that specifically binds to  Clostridium difficile  toxin A or to toxin B, or that binds to, or cross reacts with, both toxin A and B, wherein:
 a) the isolated antibody or antigen-binding fragment thereof that specifically binds toxin A of  Clostridium difficile  comprises the three heavy chain complementarity determining regions (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 98, 114, 130, 146 and 162; and the three light chain complementarity determining regions (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) amino acid sequence selected from the group consisting of SEQ ID NOs: 10, 106, 122, 138, 154 and 170;   b) the isolated antibody or antigen-binding fragment thereof that specifically binds toxin B of  Clostridium difficile  comprises the HCDR1, HCDR2 and HCDR3 contained within a HCVR amino acid sequence selected from the group consisting of SEQ ID NOs: 178, 194, 210, 226, 242, 258, 274, 290, 306, 322, 338 and 354; and the LCDR1, LCDR2 and LCDR3 contained within a LCVR amino acid sequence selected from the group consisting of SEQ ID NOs: 186, 202, 218, 234, 250, 266, 282, 298, 314, 330, 346 and 362; and   c) the isolated antibody or antigen-binding fragment that binds to, or cross reacts with both toxin A and toxin B of  Clostridium difficile  comprises the HCDR1, HCDR2 and HCDR3 contained within a HCVR amino acid sequence selected from the group consisting of SEQ ID NOs: 18, 34, 50, 66 and 82; and the LCDR1, LCDR2 and LCDR3 contained within a LCVR amino acid sequence selected from the group consisting of SEQ ID NOs: 26, 42, 58, 74 and 90.   
     
     
         2 . The isolated antibody or antigen-binding fragment thereof of  claim 1  that specifically binds toxin A of  Clostridium difficile , wherein the antibody, or antigen-binding fragment thereof comprises: (a) a HCVR having an amino acid sequence of SEQ ID NO: 146; and (b) a LCVR having an amino acid sequence of SEQ ID NO: 154. 
     
     
         3 . The isolated antibody or antigen-binding fragment thereof of  claim 1  that specifically binds toxin A of  Clostridium difficile , wherein the antibody, or antigen-binding fragment thereof comprises a HCVR/LCVR amino acid sequence pair selected from the group consisting of SEQ ID NOs: 2/10, 98/106, 114/122, 130/138, 146/154 and 162/170. 
     
     
         4 . The isolated antibody of  claim 1 , or an antigen-binding fragment thereof that specifically binds toxin A of  Clostridium difficile , wherein the antibody comprises:
 (a) a HCDR1 domain having an amino acid sequence selected from SEQ ID NOs: 4, 100, 116, 132, 148 and 164;   (b) a HCDR2 domain having an amino acid sequence selected from SEQ ID NOs: 6, 102, 118, 134, 150 and 166;   (c) a HCDR3 domain having an amino acid sequence selected from SEQ ID NOs: 8, 104, 120, 136, 152 and 168;   (d) a LCDR1 domain having an amino acid sequence selected from SEQ ID NOs: 12, 108, 124, 140, 156, and 172;   (e) a LCDR2 domain having an amino acid sequence selected from SEQ ID NOs: 14, 110, 126, 142, 158 and 174; and   (f) a LCDR3 domain having an amino acid sequence selected from SEQ ID NOs: 16, 112, 128, 144, 160 and 176.   
     
     
         5 . The isolated antibody of  claim 4 , or an antigen-binding fragment thereof that specifically binds toxin A of  Clostridium difficile , wherein the antibody comprises:
 (a) a HCDR1 domain having an amino acid sequence of SEQ ID NO: 148;   (b) a HCDR2 domain having an amino acid sequence of SEQ ID NO: 150;   (c) a HCDR3 domain having an amino acid sequence of SEQ ID NO: 152;   (d) a LCDR1 domain having an amino acid sequence of SEQ ID NO: 156;   (e) a LCDR2 domain having an amino acid sequence of SEQ ID NO: 158; and   (f) a LCDR3 domain having an amino acid sequence of SEQ ID NO: 160.   
     
     
         6 . The isolated antibody of  claim 1 , or an antigen-binding fragment thereof that specifically binds  Clostridium difficile  toxin B, wherein the antibody comprises: (a) a HCVR having the amino acid sequence of SEQ ID NO: 274; and (b) a LCVR having the amino acid sequence of SEQ ID NO: 282. 
     
     
         7 . The isolated antibody of  claim 1 , or an antigen-binding fragment thereof that specifically binds  Clostridium difficile  toxin B, wherein the isolated antibody or antigen-binding fragment thereof comprises: a HCVR/LCVR amino acid sequence pair selected from the group consisting of SEQ ID NOs: 178/186, 194/202, 210/218, 226/234, 242/250, 258/266, 274/282, 290/298, 306/314, 322/330, 338/346 and 354/362. 
     
     
         8 . The isolated antibody of  claim 1 , or an antigen-binding fragment thereof that specifically binds  Clostridium difficile  toxin B, wherein the antibody comprises:
 (a) a HCDR1 domain having an amino acid sequence selected from the group consisting of SEQ ID NOs: 180, 196, 212, 228, 244, 260, 276, 292, 308, 324, 340 and 356;   (b) a HCDR2 domain having an amino acid sequence selected from the group consisting of SEQ ID NOs: 182, 198, 214, 230, 246, 262, 278, 294, 310, 326, 342 and 358;   (c) a HCDR3 domain having an amino acid sequence selected from the group consisting of SEQ ID NOs: 184, 200, 216, 232, 248, 264, 280, 296, 312, 328, 344 and 360;   (d) a LCDR1 domain having an amino acid sequence selected from the group consisting of SEQ ID NOs: 188, 204, 220, 236, 252, 268, 284, 300, 316, 332, 348 and 364;   (e) a LCDR2 domain having an amino acid sequence selected from the group consisting of SEQ ID NOs: 190, 206, 222, 238, 254, 270, 286, 302, 318, 334, 350 and 366; and   (f) a LCDR3 domain having an amino acid sequence selected from the group consisting of SEQ ID NOs: 192, 208, 224, 240, 256, 272, 288, 304, 320, 336, 352 and 368.   
     
     
         9 . The isolated antibody of  claim 8 , or an antigen-binding fragment thereof that specifically binds  Clostridium difficile  toxin B, wherein the antibody comprises:
 (a) a HCDR1 domain having an amino acid sequence selected from the group consisting of SEQ ID NO: 276,   (b) a HCDR2 domain having an amino acid sequence selected from the group consisting of SEQ ID NOs: 278;   (c) a HCDR3 domain having an amino acid sequence selected from the group consisting of SEQ ID NOs: 280;   (d) a LCDR1 domain having an amino acid sequence selected from the group consisting of SEQ ID NOs: 284;   (e) a LCDR2 domain having an amino acid sequence selected from the group consisting of SEQ ID NOs: 286; and   (f) a LCDR3 domain having an amino acid sequence selected from the group consisting of SEQ ID NOs: 288.   
     
     
         10 . The isolated antibody of  claim 1 , or an antigen-binding fragment thereof that binds to, or cross reacts with both toxin A and toxin B of  Clostridium difficile , wherein the antibody comprises (a) a HCVR having an amino acid sequence selected from the group consisting of SEQ ID NOs: 18 and 34; and (b) a LCVR having an amino acid sequence selected from the group consisting of SEQ ID NOs: 26 and 42. 
     
     
         11 . The isolated antibody of  claim 1 , or an antigen-binding fragment thereof that binds to, or cross reacts with both toxin A and toxin B of  Clostridium difficile , wherein the antibody or antigen-binding fragment thereof comprises a HCVR/LCVR amino acid sequence pair selected from the group consisting of SEQ ID NOs: 18/26, 34/42, 50/58, 66/74 and 82/90. 
     
     
         12 . The isolated antibody of  claim 11 , or an antigen-binding fragment thereof that binds to or cross reacts with both toxin A and toxin B of  Clostridium difficile , wherein the antibody or antigen-binding fragment thereof comprises a HCVR/LCVR amino acid sequence pair selected from the group consisting of SEQ ID NOs: 18/26 and 34/42. 
     
     
         13 . The isolated antibody of  claim 1 , or an antigen-binding fragment thereof that binds to, or cross reacts with both toxin A and toxin B of  Clostridium difficile , wherein the antibody comprises:
 (a) a HCDR1 domain having an amino acid sequence selected from SEQ ID NOs: 20, 36, 52, 68, and 84;   (b) a HCDR2 domain having an amino acid sequence selected from SEQ ID NOs: 22, 38, 54, 70 and 86;   (c) a HCDR3 domain having an amino acid sequence selected from SEQ ID NOs: 24, 40, 56, 72 and 88;   (d) a LCDR1 domain having an amino acid sequence selected from SEQ ID NOs: 28, 44, 60, 76 and 92;   (e) a LCDR2 domain having an amino acid sequence selected from SEQ ID NOs: 30, 46, 62, 78 and 94; and   (f) a LCDR3 domain having an amino acid sequence selected from SEQ ID NOs: 32, 48, 64, 80 and 96.   
     
     
         14 . The isolated antibody of  claim 13 , or an antigen-binding fragment thereof that binds to, or cross reacts with both toxin A and toxin B of  Clostridium difficile , wherein the isolated antibody or antigen-binding fragment comprises:
 (a) a HCDR1 domain having an amino acid sequence selected from SEQ ID NOs: 20, and 36;   (b) a HCDR2 domain having an amino acid sequence selected from SEQ ID NOs: 22, and 38;   (c) a HCDR3 domain having an amino acid sequence selected from SEQ ID NOs: 24, and 40;   (d) a LCDR1 domain having an amino acid sequence selected from SEQ ID NOs: 28, and 44;   (e) a LCDR2 domain having an amino acid sequence selected from SEQ ID NOs: 30, and 46; and   (f) a LCDR3 domain having an amino acid sequence selected from SEQ ID NOs: 32, and 48.   
     
     
         15 . A pharmaceutical composition comprising one or more antibodies of  claim 1  and a pharmaceutically acceptable carrier or diluent. 
     
     
         16 . The pharmaceutical composition of  claim 15 , wherein the composition comprises at least one antibody, or an antigen-binding fragment thereof that binds specifically to toxin A of  Clostridium difficile  and at least one antibody, or an antigen-binding fragment thereof that binds specifically to toxin B of  Clostridium difficile , wherein:
 a) the antibody or antigen-binding fragment thereof that binds specifically to toxin A comprises the three heavy chain complementarity determining regions (HCDR1, HCDR2 and HCDR3) contained within any one of the heavy chain variable region (HCVR) amino acid sequences selected from the group consisting of SEQ ID NOs: 2, 98, 114, 130, 146 and 162; and the three light chain complementarity determining regions (LCDR1, LCDR2 and LCDR3) contained within any one of the light chain variable region (LCVR) amino acid sequences selected from the group consisting of SEQ ID NOs: 10, 106, 122, 138, 154 and 170; and wherein   b) the antibody or antigen-binding fragment thereof that binds specifically to toxin B comprises the three heavy chain CDRs (HCDR1, HCDR2 and HCDR3) contained within any one of the HCVR amino acid sequences selected from the group consisting of SEQ ID NOs: 178, 194, 210, 226, 242, 258, 274, 290, 306, 322, 338 and 354; and the three light chain CDRs (LCDR1, LCDR2 and LCDR3) contained within any one of the LCVR amino acid sequences selected from the group consisting of SEQ ID NOs: 186, 202, 218, 234, 250, 266, 282, 298, 314, 330, 346 and 362.   
     
     
         17 . The pharmaceutical composition of  claim 16 , wherein the antibody or an antigen-binding fragment thereof that specifically binds toxin A of  Clostridium difficile  comprises a HCVR/LCVR amino acid sequence pair of SEQ ID NO: 146/154, and wherein the antibody or an antigen-binding fragment thereof that specifically binds toxin B of  Clostridium difficile  comprises a HCVR/LCVR amino acid sequence pair of SEQ ID NO: 274/282. 
     
     
         18 . The pharmaceutical composition of  claim 16 , comprising:
 a) an isolated first antibody, or antigen-binding fragment thereof that specifically binds toxin A of  Clostridium difficile , comprising a HCDR1 having the amino acid sequence of SEQ ID NO: 148, a HCDR2 having the amino acid sequence of SEQ ID NO: 150, a HCDR3 having the amino acid sequence of SEQ ID NO: 152, a LCDR1 having the amino acid sequence of SEQ ID NO: 156, a LCDR2 having the amino acid sequence of SEQ ID NO: 158, a LCDR3 having the amino acid sequence of SEQ ID NO: 160;   b) an isolated second antibody, or antigen-binding fragment thereof that specifically binds toxin B of  Clostridium difficile , comprising a HCDR1 having the amino acid sequence of SEQ ID NO: 276, a HCDR2 having the amino acid sequence of SEQ ID NO: 278, a HCDR3 having the amino acid sequence of SEQ ID NO: 280, a LCDR1 having the amino acid sequence of SEQ ID NO: 284, a LCDR2 having the amino acid sequence of SEQ ID NO: 286, a LCDR3 having the amino acid sequence of SEQ ID NO: 288; and   c) a pharmaceutically acceptable carrier or diluent.   
     
     
         19 . The pharmaceutical composition of  claim 15 , wherein the antibodies contained within the composition are effective at neutralizing toxins A and B from a hypervirulent strain of  Clostridium difficile.    
     
     
         20 . The pharmaceutical composition of  claim 19 , wherein the hypervirulent strain of  Clostridium difficile  is a BI/NAP1/027 strain. 
     
     
         21 . The pharmaceutical composition of  claim 20 , wherein the BI/NAP1/027 strain is selected from VA5, VA17, 6336 and 6443. 
     
     
         22 . A method for treating a patient suffering from a  Clostridium difficile -associated condition or disease, or for treating at least one symptom or complication associated with the condition or disease, or for preventing the development of a  Clostridium difficile -associated condition or disease in a patient at risk thereof, the method comprising administering to the patient an effective amount of the pharmaceutical composition of  claim 15 , wherein the  Clostridium difficile -associated condition or disease is either prevented, or lessened in severity and/or duration, or at least one symptom or complication associated with the condition or disease is prevented, or ameliorated, or that the frequency and/or duration of, or the severity of recurrences, or relapses with  Clostridium difficile  is reduced. 
     
     
         23 . The method of  claim 22 , wherein the at least one symptom or complication associated with the  Clostridium difficile -associated condition or disease is selected from the group consisting of anorexia, abdominal pain, abdominal bloating, diarrhea with or without bleeding, dehydration, malnutrition, pseudomembranous colitis, complete or segmental colonic resection, fever and systemic infection (sepsis), death, relapse of the  Clostridium difficile  condition or disease, and rejection of a transplanted tissue or organ. 
     
     
         24 . The method of  claim 22 , wherein the patient at risk of developing a  Clostridium difficile -associated condition or disease is selected from the group consisting of an elderly patient 65 years old), a patient who is immunocompromised due to underlying illness or due to administration of immunosuppressive therapeutics, a patient who has some underlying medical condition that may pre-dispose them to acquiring a  Clostridium difficile  infection, a patient hospitalized for an extended period of time (at least one week), a patient who has been treated for an extended period of time(≧14 days) with broad spectrum antibiotics, a cancer patient, a transplant patient, and a patient on therapy with agents such as but not limited to a proton pump inhibitor, or histamine H2 receptor inhibitor that are used for treatment of gastrointestinal diseases or conditions to reduce or treat gastric acidity, gastroesophageal reflux disease (GERD), stomach and small intestine ulcers, or heartburn. 
     
     
         25 . The method of  claim 24 , wherein the cancer patient is undergoing treatment with an anti-cancer drug, or undergoing radiotherapy to treat a cancer. 
     
     
         26 . The method of  claim 24 , wherein the transplant patient is a patient receiving a hematopoietic stem cell transplant, or a solid tissue or organ transplant. 
     
     
         27 . The method of  claim 26 , wherein the transplant patient is being treated with an immunosuppressive drug, or any transplant rejection drug, or who is undergoing treatment with a drug regimen to prevent tissue or organ graft rejection following the transplant. 
     
     
         28 . The method of  claim 22 , wherein the pharmaceutical composition is administered to the patient in combination with a second therapeutic agent. 
     
     
         29 . The method of  claim 28 , wherein the second therapeutic agent is selected from the group consisting of a toxoid, a  Clostridium difficile  vaccine, an antibiotic, another different antibody to  Clostridium difficile  toxin A and/or B, and any other palliative therapy useful for ameliorating at least one symptom associated with a  Clostridium difficile -associated condition or disease. 
     
     
         30 . The method of  claim 29 , wherein the at least one symptom or complication associated with the  Clostridium difficile -associated condition or disease is selected from the group consisting of anorexia, abdominal pain, abdominal bloating, diarrhea with or without bleeding, dehydration, malnutrition, pseudomembranous colitis, complete or segmental colonic resection, fever and systemic infection (sepsis), death, relapse of the  Clostridium difficile  condition or disease, and rejection of a transplanted tissue or organ. 
     
     
         31 . An isolated antibody that interacts with, or binds to, an epitope within amino acid residues 468-863 of the carboxy terminal receptor binding domain of toxin A produced by  Clostridium difficile , or an antigen binding fragment thereof, wherein the carboxy terminal receptor binding domain of toxin A comprises the amino acid sequence of SEQ ID NO: 375. 
     
     
         32 . An isolated antibody that interacts with or binds to an epitope within the carboxy terminal receptor binding domain of toxin A produced by  Clostridium difficile , or an antigen binding fragment thereof, wherein the epitope is selected from the group consisting of residues 468-488 of SEQ ID NO: 375, residues 510-530 of SEQ ID NO: 375, residues 602-610 of SEQ ID NO: 375, residues 644-703 of SEQ ID NO: 375, residues 724-794 of SEQ ID NO: 375, residues 799-814 of SEQ ID NO: 375 and residues 858-863 of SEQ ID NO: 375. 
     
     
         33 . The isolated antibody of  claim 32 , comprising the HCVR/LCVR amino acid sequence pair of SEQ ID NOs: 146/154. 
     
     
         34 . A pharmaceutical composition comprising the isolated antibody of  claim 32  and a second isolated antibody that interacts with, or binds to toxin B of  Clostridium difficile  and a pharmaceutically acceptable carrier or diluent. 
     
     
         35 . The pharmaceutical composition of  claim 34 , wherein the second antibody that interacts with or binds to toxin B of  Clostridium difficile  comprises the HCVR/LCVR amino acid sequence pair of SEQ ID NOs: 274/282.