Interferon-gamma-binding molecules for treating septic shock, cachexia, immune diseases and skin disorders

Abstract

The present invention concerns molecules which bind and neutralize the cytokine interferon-gamma. More specifically, the present invention relates to sheep-derived antibodies and engineered antibody constructs, such as humanized single-chain Fv fragments, chimeric antibodies, diabodies, triabodies, tetravalent antibodies, peptabodies and hexabodies which can be used to treat diseases wherein interferon-gamma activity is pathogenic. Examples of such diseases are: septic shock, cachexia, multiple sclerosis and psoriasis.

1 . A molecule which binds and neutralizes interferon-gamma and which is chosen from the group consisting of: 
 a scFv comprising the humanized variable domain of the monoclonal antibody D9D10    a chimeric antibody comprising the humanized variable domain of the monoclonal antibody D9D10    a diabody comprising the humanized variable domain of the monoclonal antibody D9D10 and    a multivalent antibody comprising the humanized variable domain of the monoclonal antibody D9D10.    
     
     
         2 . The molecule according to  claim 1 , wherein said multivalent antibody is chosen from the group consisting of triabodies and tetravalent antibodies.  
     
     
         3 . The molecule according to  claim 2 , wherein said triabody or tetravalent antibody, comprise 3 and 4 variable domains, respectively, of several anti-interferon-gamma antibodies.  
     
     
         4 . The molecule according to  claim 2 , wherein said triabody comprises 3 identical variable domains of an anti-interferon-gamma antibody.  
     
     
         5 . The molecule according to  claim 2 , wherein said triabody comprises 3 identical D9D10 scFv's.  
     
     
         6 . The molecule according to  claim 2 , wherein said triabody comprises 3 identical humanized D9D10 scFv's.  
     
     
         7 . The molecule according to  claim 2 , wherein said tetravalent antibody comprises 4 identical domains of an anti-interferon-gamma antibody.  
     
     
         8 . The molecule according to  claim 2 , wherein said tetravalent antibody comprises 4 identical D9D10 scFv's.  
     
     
         9 . The molecule according to  claim 2 , wherein said tetravalent antibody comprises 4 identical humanized D9D10 scFv's.  
     
     
         10 . The molecule according to  claim 2 , wherein said tetravalent antibody comprises a full-size humanized D9D10 antibody.  
     
     
         11 . A pharmaceutical composition comprising a molecule according to  claim 1  or a mixture of said molecules in a pharmaceutically acceptable excipient.  
     
     
         12 . A method for neutralizing interferon-gamma activity in a mammal comprising administering to the mammal a pharmaceutically effective amount of a molecule that binds and neutralizes interferon-gamma, said molecule selected from the group consisting of: 
 a scFv comprising the humanized variable domain of the monoclonal antibody D9D10    a chimeric antibody comprising the humanized variable domain of the monoclonal antibody D9D10    a diabody comprising the humanized variable domain of the monoclonal antibody D9D10 and    a multivalent antibody comprising the humanized variable domain of the monoclonal antibody D9D10.    
     
     
         13 . The method according to  claim 12  wherein the pharmaceutically effective amount of the molecule that binds and neutralizes interferon-gamma is administered to prevent or treating septic shock, cachexia, autoimmune disease(s), and/or skin disorder(s).  
     
     
         14 . The method of  claim 13  wherin the autoimmune disease is multiple sclerosis, Crohn's disease, or rheumatoid arthritis.  
     
     
         15 . The method of  claim 13  wherein the skin disorder is bullous, inflammatory, or neoplastic dermatosis.  
     
     
         16 . The method of  claim 12  wherein said multivalent antibody is chosen from the group consisting of triabodies and tetravalent antibodies.  
     
     
         17 . The method according to  claim 12  wherein said triabody or tetravalent antibody, comprise 3 and 4 variable domains, respectively, of several anti-interferon-gamma antibodies.  
     
     
         18 . The method according to  claim 12  wherein said triabody comprises 3 identical variable domains of an anti-interferon-gamma antibody.  
     
     
         19 . The method according to  claim 12  wherein said triabody comprises 3 identical D9D10 scFv's.  
     
     
         20 . The method according to  claim 12  wherein said triabody comprises 3 identical humanized D9D10 scFv's.  
     
     
         21 . The method according to  claim 12  wherein said tetravalent antibody comprises 4 identical domains of an anti-interferon-gamma antibody.  
     
     
         22 . The method according to  claim 12  wherein said tetravalent antibody comprises 4 identical D9D10 scFv's.  
     
     
         23 . The method according to  claim 12  wherein said tetravalent antibody comprises 4 identical humanized D9D10 scFv's.  
     
     
         24 . The method according to  claim 12  wherein said tetravalent antibody comprises a full-size humanized D9D10 antibody.  
     
     
         25 . A method for determining interferon gamma levels in a sample comprising: 
 a) contacting the biological sample to be analysed for the presence of interferon-gamma with a molecule according to  claim 1;     b) detecting the immunological complex formed between interferon-gamma and said molecule.