Neutralising antibodies to the major exotoxins tcda and tcdb of clostridium difficile

Abstract

This present invention describes the derivation and selection of antibodies capable of neutralising the major exotoxins; TcdA and TcdB of Clostridium difficile. The invention also describes novel neutralisation and antigen binding properties of individual Mabs and mixtures thereof.

1 - 41 . (canceled) 
     
     
         42 . A pharmaceutical composition comprising one or more monoclonal antibodies specific to antigen TcdB1234 wherein the antibody has high affinity of 600 pM or less for the target antigen TcdB1234 and said one or more monoclonal antibodies are independently selected from:
 i) a heavy chain wherein the variable domain of the heavy chain comprises a CDR having the sequence given in SEQ ID NO:124 for CDR-H1, a CDR having the sequence given in SEQ ID NO: 125 for CDR-H2 and a CDR having the sequence given in SEQ ID NO: 126 for CDR-H3, and a light chain wherein the variable domain of the light chain comprises a CDR having the sequence given in SEQ ID NO:121 for CDR-L1, a CDR having the sequence given in in SEQ ID NO:122 for CDR-L2 and a CDR having the sequence given in SEQ ID NO:123 for CDR-L3;   ii) a heavy chain wherein the variable domain of the heavy chain comprises a CDR having the sequence given in SEQ ID NO:154 for CDR-H1, a CDR having the sequence given in SEQ ID NO:155 for CDR-H2 and a CDR having the sequence given in SEQ ID NO:156 for CDR-H3, and a light chain wherein the variable domain of the light chain comprises a CDR having the sequence given in SEQ ID NO:151 for CDR-L1, a CDR having the sequence given in in SEQ ID NO:152 for CDR-L2 and a CDR having the sequence given in SEQ ID NO:153 for CDR-L3;   iii) a heavy chain wherein the variable domain of the heavy chain comprises a CDR having the sequence given in SEQ ID NO:144 for CDR-H1, a CDR having the sequence given in SEQ ID NO:145 for CDR-H2 and a CDR having the sequence given in SEQ ID NO:146 for CDR-H3, and a light chain wherein the variable domain of the light chain comprises a CDR having the sequence given in SEQ ID NO:141 for CDR-L1, a CDR having the sequence given in in SEQ ID NO:142 for CDR-L2 and a CDR having the sequence given in SEQ ID NO:143 for CDR-L3;   iv) a heavy chain wherein the variable domain of the heavy chain comprises a CDR having the sequence given in SEQ ID NO:164 for CDR-H1, a CDR having the sequence given in SEQ ID NO:165 for CDR-H2 and a CDR having the sequence given in SEQ ID NO:166 for CDR-H3, and a light chain wherein the variable domain of the light chain comprises a CDR having the sequence given in SEQ ID NO:161 for CDR-L1, a CDR having the sequence given in in SEQ ID NO:162 for CDR-L2 and a CDR having the sequence given in SEQ ID NO:163 for CDR-L3;   v) a heavy chain wherein the variable domain of the heavy chain comprises a CDR having the sequence given in SEQ ID NO:64 for CDR-H1, a CDR having the sequence given in SEQ ID NO: 65 for CDR-H2 and a CDR having the sequence given in SEQ ID NO: 66 for CDR-H3, and a light chain wherein the variable domain of the light chain comprises a CDR having the sequence given in SEQ ID NO:61 for CDR-L1, a CDR having the sequence given in in SEQ ID NO:62 for CDR-L2 and a CDR having the sequence given in SEQ ID NO:63 for CDR-L3;   vi) a heavy chain wherein the variable domain of the heavy chain comprises a CDR having the sequence given in SEQ ID NO:74 for CDR-H1, a CDR having the sequence given in SEQ ID NO: 75 for CDR-H2 and a CDR having the sequence given in SEQ ID NO: 76 for CDR-H3, and a light chain wherein the variable domain of the light chain comprises a CDR having the sequence given in SEQ ID NO:71 for CDR-L1, a CDR having the sequence given in in SEQ ID NO:72 for CDR-L2 and a CDR having the sequence given in SEQ ID NO:73 for CDR-L3;   vii) a heavy chain wherein the variable domain of the heavy chain comprises a CDR having the sequence given in SEQ ID NO:84 for CDR-H1, a CDR having the sequence given in SEQ ID NO: 85 for CDR-H2 and a CDR having the sequence given in SEQ ID NO: 86 for CDR-H3, and a light chain wherein the variable domain of the light chain comprises a CDR having the sequence given in SEQ ID NO:81 for CDR-L1, a CDR having the sequence given in in SEQ ID NO:82 for CDR-L2 and a CDR having the sequence given in SEQ ID NO:83 for CDR-L3;   viii) a heavy chain wherein the variable domain of the heavy chain comprises a CDR having the sequence given in SEQ ID NO:134 for CDR-H1, a CDR having the sequence given in SEQ ID NO: 135 for CDR-H2 and a CDR having the sequence given in SEQ ID NO: 136 for CDR-H3, and a light chain wherein the variable domain of the light chain comprises a CDR having the sequence given in SEQ ID NO:131 for CDR-L1, a CDR having the sequence given in in SEQ ID NO:132 for CDR-L2 and a CDR having the sequence given in SEQ ID NO:133 for CDR-L3.   
     
     
         43 . A pharmaceutical composition according to  claim 42 , wherein said one or more monoclonal antibodies are independently selected from:
 i) a heavy chain wherein the variable domain of the heavy chain comprises a CDR having the sequence given in SEQ ID NO:124 for CDR-H1, a CDR having the sequence given in SEQ ID NO: 125 for CDR-H2 and a CDR having the sequence given in SEQ ID NO: 126 for CDR-H3, and a light chain wherein the variable domain of the light chain comprises a CDR having the sequence given in SEQ ID NO:121 for CDR-L1, a CDR having the sequence given in in SEQ ID NO:122 for CDR-L2 and a CDR having the sequence given in SEQ ID NO:123 for CDR-L3;   ii) a heavy chain wherein the variable domain of the heavy chain comprises a CDR having the sequence given in SEQ ID NO:154 for CDR-H1, a CDR having the sequence given in SEQ ID NO:155 for CDR-H2 and a CDR having the sequence given in SEQ ID NO:156 for CDR-H3, and a light chain wherein the variable domain of the light chain comprises a CDR having the sequence given in SEQ ID NO:151 for CDR-L1, a CDR having the sequence given in in SEQ ID NO:152 for CDR-L2 and a CDR having the sequence given in SEQ ID NO:153 for CDR-L3;   iii) a heavy chain wherein the variable domain of the heavy chain comprises a CDR having the sequence given in SEQ ID NO:144 for CDR-H1, a CDR having the sequence given in SEQ ID NO:145 for CDR-H2 and a CDR having the sequence given in SEQ ID NO:146 for CDR-H3, and a light chain wherein the variable domain of the light chain comprises a CDR having the sequence given in SEQ ID NO:141 for CDR-L1, a CDR having the sequence given in in SEQ ID NO:142 for CDR-L2 and a CDR having the sequence given in SEQ ID NO:143 for CDR-L3; and   iv) a heavy chain wherein the variable domain of the heavy chain comprises a CDR having the sequence given in SEQ ID NO:164 for CDR-H1, a CDR having the sequence given in SEQ ID NO:165 for CDR-H2 and a CDR having the sequence given in SEQ ID NO:166 for CDR-H3, and a light chain wherein the variable domain of the light chain comprises a CDR having the sequence given in SEQ ID NO:161 for CDR-L1, a CDR having the sequence given in in SEQ ID NO:162 for CDR-L2 and a CDR having the sequence given in SEQ ID NO:163 for CDR-L3.   
     
     
         44 . A pharmaceutical composition according to  claim 42 , wherein a monoclonal antibody which specifically binds TcdB1234 comprises a heavy chain wherein the variable domain of the heavy chain comprises a CDR having the sequence given in SEQ ID NO:124 for CDR-H1, a CDR having the sequence given in SEQ ID NO: 125 for CDR-H2 and a CDR having the sequence given in SEQ ID NO: 126 for CDR-H3, and a light chain wherein the variable domain of the light chain comprises a CDR having the sequence given in SEQ ID NO:121 for CDR-L1, a CDR having the sequence given in in SEQ ID NO:122 for CDR-L2 and a CDR having the sequence given in SEQ ID NO:123 for CDR-L3. 
     
     
         45 . A pharmaceutical composition according to  claim 44 , wherein the monoclonal antibody has a heavy chain comprising the sequence given in SEQ ID NO:129 and a light chain comprising the sequence given in SEQ ID NO:127. 
     
     
         46 . A pharmaceutical composition according to  claim 42 , wherein a monoclonal antibody which specifically binds TcdB1234 comprising a heavy chain wherein the variable domain of the heavy chain comprises a CDR having the sequence given in SEQ ID NO:154 for CDR-H1, a CDR having the sequence given in SEQ ID NO:155 for CDR-H2 and a CDR having the sequence given in SEQ ID NO:156 for CDR-H3, and a light chain wherein the variable domain of the light chain comprises a CDR having the sequence given in SEQ ID NO:151 for CDR-L1, a CDR having the sequence given in in SEQ ID NO:152 for CDR-L2 and a CDR having the sequence given in SEQ ID NO:153 for CDR-L3. 
     
     
         47 . A pharmaceutical composition according to  claim 46 , wherein the monoclonal antibody has a heavy chain comprising the sequence given in SEQ ID NO:159 and a light chain comprising the sequence given in SEQ ID NO:157. 
     
     
         48 . A pharmaceutical composition according to  claim 42 , wherein the monoclonal antibody has a heavy chain comprising the sequence given in SEQ ID NO:149 and a light chain comprising the sequence given in SEQ ID NO:147. 
     
     
         49 . A pharmaceutical composition according to  claim 42 , wherein the monoclonal antibody has a heavy chain comprising the sequence given in SEQ ID NO:169 and a light chain comprising the sequence given in SEQ ID NO:167. 
     
     
         50 . A pharmaceutical composition according to  claim 42 , wherein at least one of said monoclonal antibodies is a neutralizing antibody which is effective against ribotypes 003. 
     
     
         51 . A pharmaceutical composition according to  claim 42 , further comprising two or more antibodies specific to TcdB. 
     
     
         52 . A pharmaceutical composition according to  claim 42 , further comprising one or more antibodies specific to TcdA. 
     
     
         53 . A pharmaceutical composition according to  claim 52 , wherein the antibody which specifically binds TcdA comprises a heavy chain sequence given in SEQ ID NO: 19 and a light chain sequence given in SEQ ID NO: 17. 
     
     
         54 . A pharmaceutical composition according to  claim 52 , wherein the antibody which specifically binds TcdA comprises a heavy chain sequence given in SEQ ID NO: 29 and a light chain sequence given in SEQ ID NO: 27. 
     
     
         55 . A pharmaceutical composition according to  claim 52 , wherein the antibody which specifically binds TcdA comprises a heavy chain sequence given in SEQ ID NO: 49 and a light chain sequence given in SEQ ID NO: 47. 
     
     
         56 . A pharmaceutical composition according to  claim 52 , wherein the antibody which specifically binds TcdA comprises a heavy chain sequence given in SEQ ID NO: 59 and a light chain sequence given in SEQ ID NO: 57. 
     
     
         57 . A pharmaceutical composition according to  claim 52 , wherein the antibody which specifically binds TcdA comprises a heavy chain sequence given in SEQ ID NO: 39 and a light chain sequence given in SEQ ID NO: 37. 
     
     
         58 . A pharmaceutical composition according to  claim 52 , wherein the antibody which specifically binds TcdA comprises a heavy chain sequence given in SEQ ID NO: 9 and a light chain sequence given in SEQ ID NO: 7. 
     
     
         59 . A pharmaceutical composition according to  claim 52 , which further comprises a pharmaceutically acceptable excipient. 
     
     
         60 . A method of treatment or prophylaxis of  Clostridium difficile  infection or complications therefrom comprising administering a pharmaceutical composition as defined in  claim 42  to a patient in need thereof. 
     
     
         61 . A method according to  claim 60 , wherein the composition is co-administrated with a compound selected the group comprising metronidazole, vancomycin, clindamycin, fidaxomicin and combinations thereof.