Humanized anti-CD3 specific antibodies

Abstract

Novel aglycosylated antibodies having a binding affinity for the CD3 antigen complex are of value for use in therapy, particularly in immunosuppression.

1 . An aglycosylated IgG antibody having a binding affinity for the CD3 antigen complex.  
     
     
         2 . An aglycosylated antibody according to  claim 1 , which has a binding affinity for the human CD3 antigen complex.  
     
     
         3 . An aglycosylated antibody according to  claim 2 , in which at least one CDR is selected from the amino acid sequences:  
       
         
           
                 
                 
                 
               
                     
                 
                   (a) 
                   Ser-Phe-Pro-Met-Ala, 
                     
                 
                     
                 
                   (b) 
                   Thr-Ile-Ser-Thr-Ser-Gly-Gly-Arg-Thr-Tyr-Tyr- 
                 
                     
                   Arg-Asp-Ser-Val-Lys-Gly, 
                 
                     
                 
                   (c) 
                   Phe-Arg-Gln-Tyr-Ser-Gly-Gly-Phe-Asp-Tyr, 
                 
                     
                 
                   (d) 
                   Thr-Leu-Ser-Ser-Gly-Asn-Ile-Glu-Asn-Asn-Tyr- 
                 
                     
                   Val-His, 
                 
                     
                 
                   (e) 
                   Asp-Asp-Asp-Lys-Arg-Pro-Asp, 
                 
                     
                 
                   (f) 
                   His-Ser-Tyr-Val-Ser-Ser-Phe-Asn-Val, 
                 
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         and conservatively modified variants thereof.  
       
     
     
         4 . An aglycosylated antibody according to  claim 2 , which has a heavy chain with at least one CDR selected from the amino acid sequences:  
       
         
           
                 
                 
                 
               
                     
                 
                   (a) 
                   Ser-Phe-Pro-Met-Ala, 
                     
                 
                     
                 
                   (b) 
                   Thr-Ile-Ser-Thr-Ser-Gly-Gly-Arg-Thr-Tyr-Tyr- 
                 
                     
                   Arg-Asp-Ser-Val-Lys-Gly, 
                 
                     
                 
                   (c) 
                   Phe-Arg-Gln-Tyr-Ser-Gly-Gly-Phe-Asp-Tyr, 
                 
                     
                 
             
                
                
                
                
                
                
                
                
               
            
           
         
         and conservatively modified variants thereof, and/or a light chain with at least one CDR selected from the amino acid sequences:  
         
           
             
                   
                   
                   
                 
                       
                   
                     (d) 
                     Thr-Leu-Ser-Ser-Gly-Asn-Ile-Glu-Asn-Asn-Tyr- 
                       
                   
                       
                     Val-His, 
                   
                       
                   
                     (e) 
                     Asp-Asp-Asp-Lys-Arg-Pro-Asp, 
                   
                       
                   
                     (f) 
                     His-Ser-Tyr-Val-Ser-Ser-Phe-Asn-Val, 
                   
                       
                   
               
                  
                  
                  
                  
                  
                  
                  
                  
                 
              
             
           
         
         and conservatively modified variants thereof.  
       
     
     
         5 . An aglycosylated antibody according to  claim 2 , which has a heavy chain with three CDRs comprising the amino acid sequences:  
       
         
           
                 
                 
                 
               
                     
                 
                   (a) 
                   Ser-Phe-Pro-Met-Ala, 
                     
                 
                     
                 
                   (b) 
                   Thr-Ile-Ser-Thr-Ser-Gly-Gly-Arg-Thr-Tyr-Tyr- 
                 
                     
                   Arg-Asp-Ser-Val-Lys-Gly, 
                 
                     
                 
                   (c) 
                   Phe-Arg-Gln-Tyr-Ser-Gly-Gly-Phe-Asp-Tyr, 
                 
                     
                 
             
                
                
                
                
                
                
                
                
               
            
           
         
         or conservatively modified variants thereof, and a light chain with three CDRs comprising the amino acid sequences:  
         
           
             
                   
                   
                   
                 
                       
                   
                     (d) 
                     Thr-Leu-Ser-Ser-Gly-Asn-Ile-Glu-Asn-Asn-Tyr- 
                       
                   
                       
                     Val-His, 
                   
                       
                   
                     (e) 
                     Asp-Asp-Asp-Lys-Arg-Pro-Asp, 
                   
                       
                   
                     (f) 
                     His-Ser-Tyr-Val-Ser-Ser-Phe-Asn-Val, 
                   
                       
                   
               
                  
                  
                  
                  
                  
                  
                  
                  
                 
              
             
           
         
         or conservatively modified variants thereof, the heavy chain CDRs being arranged in the order (a), (b), (c) in the leader→constant domain direction and the light chain CDRs being arranged in the order (d), (e), (f) in the leader→constant domain direction.  
       
     
     
         6 . An aglycosylated antibody according to any of claims  claim 1 , in which the variable domain framework regions are of or are derived from those of rat or mouse origin.  
     
     
         7 . An aglycosylated antibody according to any of claims  claim 1 , in which the CDRs are of different origin to the variable framework region.  
     
     
         8 . An aglycosylated antibody according to  claim 7 , in which the variable domain framework regions are of or are derived from those of human origin.  
     
     
         9 . An aglycosylated antibody according to  claim 8 , in which the heavy chain variable domain framework region reading from in the leader→constant domain direction comprises 
 Glu-Val-Gln-Leu-Leu-Glu-Ser-Gly-Gly-Gly-Leu-Val-Gln-Pro-Gly-Gly-Ser-Leu-Arg-Leu-Ser-Cys-Ala-Ala-Ser-Gly-Phe-Thr-Phe-Ser-/CDR/- Trp-Val-Arg-Gln-Ala-Pro-Gly-Lys-Gly-Leu-Glu-Trp-Val-Ser-/CDR/-Arg-Phe-Thr-lle-Ser-Arg-Asp-Asn-Ser-Lys-Asn-Thr-Leu-Tyr-Leu-Gln- Met-Asn-Ser-Leu-Arg-Ala-Glu-Asp-Thr-Ala-Val-Tyr-Tyr-Cys-Ala-Lys-/CDR/-Trp-Gly-Gln-Gly-Thr-Leu-Val-Thr-Val-Ser-Ser, CDR indicating the presence of a CDR of which at least one is (a), (b) or (c) or a conservatively modified variant thereof.    
     
     
         10 . An aglycosylated antibody according to  claim 8 , in which the light chain variable domain framework region reading in the leader→constant domain direction comprises  
       Asp-Phe-Met-Leu-Thr-Gln-Pro-His-Ser-Val-Ser-Glu-Ser-Pro-Gly-Lys-Thr-Val-Ile-Ile-Ser- Cys-/CDR/-Trp-Tyr-Gln-Gln-Arg-Pro-Gly-Arg-Ala-Pro-Thr-Thr-Val-lle-Phe-/CDR/-Gly- Val-Pro-Asp-Arg-Phe-Ser-Gly-Ser-lle-Asp-Arg-Ser-Ser-Asn-Ser-Ala-Ser-Leu-Thr-lle- Ser-Gly-Leu-Gln-Thr-Glu-Asp-Glu-Ala-Asp-Tyr-Tyr-Cys-/CDR/-Phe-Gly-Gly-Gly-Thr- Lys-Leu-Thr-Val-Leu-Gly-Gln-Pro-Lys-Ala-Ala-Pro-Ser-Val-Thr-Leu-Phe-Pro-Pro-Ser-Ser-Glu-Glu-Leu-Gln, CDR indicating the presence of a CDR of which at least one is (d), (e) or (f) or a conservatively modified variant thereof.  
     
     
         11 . An aglycosylated antibody according to  claim 9  having a heavy chain variable domain which comprises  
       
         
           
                 
                 
               
                     
                 
                   Glu-Val-Gln-Leu-Leu-Glu-Ser-Gly-Gly-Gly-Leu-Val- 
                     
                 
                     
                 
                   Gln-Pro-Gly-Gly-Ser-Leu-Arg-Leu-Ser-Cys-Ala-Ala- 
                 
                     
                 
                   Ser-Gly-Phe-Thr-Phe-Ser-Ser-Phe-Pro-Met-Ala-Trp- 
                 
                     
                 
                   Val-Arg-Gln-Ala-Pro-Gly-Lys-Gly-Leu-Glu-Trp-Val- 
                 
                     
                 
                   Ser-Thr-Ile-Ser-Thr-Ser-Gly-Gly-Arg-Thr-Tyr-Tyr- 
                 
                     
                 
                   Arg-Asp-Ser-Val-Lys-Gly-Arg-Phe-Thr-Ile-Ser-Arg- 
                 
                     
                 
                   Asp-Asn-Ser-Lys-Asn-Thr-Leu-Tyr-Leu-Gln-Met-Asn- 
                 
                     
                 
                   Ser-Leu-Arg-Ala-Glu-Asp-Thr-Ala-Val-Tyr-Tyr-Cys- 
                 
                     
                 
                   Ala-Lys-Phe-Arg-Gln-Tyr-Ser-Gly-Gly-Phe-Asp-Tyr- 
                 
                     
                 
                   Trp-Gly-Gln-Gly-Thr-Leu-Val-Thr-Val-Ser-Ser. 
                 
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         12 . An aglycosylated antibody according to  claim 8  having a light chain variable domain which comprises  
       
         
           
                 
                 
               
                     
                 
                   Asp-Phe-Met-Leu-Thr-Gln-Pro-His-Ser-Val-Ser-Glu- 
                     
                 
                     
                 
                   Ser-Pro-Gly-Lys-Thr-Val-Ile-Ile-Ser-Cys-Thr-Leu- 
                 
                     
                 
                   Ser-Ser-Gly-Asn-Ile-Glu-Asn-Asn-Tyr-Val-His-Trp- 
                 
                     
                 
                   Tyr-Gln-Gln-Arg-Pro-Gly-Arg-Ala-Pro-Thr-Thr-Val- 
                 
                     
                 
                   Ile-Phe-Asp-Asp-Asp-Lys-Arg-Pro-Asp-Gly-Val-Pro- 
                 
                     
                 
                   Asp-Arg-Phe-Ser-Gly-Ser-Ile-Asp-Arg-Ser-Ser-Asn- 
                 
                     
                 
                   Ser-Ala-Ser-Leu-Thr-Ile-Ser-Gly-Leu-Gln-Thr-Glu- 
                 
                     
                 
                   Asp-Glu-Ala-Asp-Tyr-Tyr-Cys-His-Ser-Tyr-Val-Ser- 
                 
                     
                 
                   Ser-Phe-Asn-Val-Phe-Gly-Gly-Gly-Thr-Lys-Leu-Thr- 
                 
                     
                 
                   Val-Leu-Gly-Gln-Pro-Lys-Ala-Ala-Pro-Ser-Val-Thr- 
                 
                     
                 
                   Leu-Phe-Pro-Pro-Ser-Ser-Glu-Glu-Leu-Gln. 
                 
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         13 . An aglycosylated antibody according to  claim 1 , in which the constant domains are of or are derived from those of rat or mouse origin.  
     
     
         14 . An aglycosylated antibody according to  claim 1 , in which the CDRs are of different origin to the constant region.  
     
     
         15 . An aglycosylated antibody according to  claim 1 , in which the constant domains are of or are derived from those of human origin.  
     
     
         16 . An aglycosylated antibody according to  claim 1 , in which the constant region is of an IgG isotype.  
     
     
         17 . An aglycosylated antibody according to  claim 15 , in which the constant region is of an IgG1 isotope.  
     
     
         18 . An aglycosylated antibody according to  claim 15 , in which asparagines residue at position 297 of each constant region heavy chain is replaced by an alternative amino acid residue.  
     
     
         19 . An aglycosylated antibody according to  claim 18 , in which the asparagine residue is replaced by an alanine residue.  
     
     
         20 . An aglycosylated antibody according to  claim 1 , in which only one of the arms thereof has an affinity for the CD3 antigen.  
     
     
         21 . An aglycosylated antibody according to  claim 20  which is monovalent.  
     
     
         22 . An aglycosylated antibody according to  claim 21 , in which one half of the antibody consists of a complete heavy chain and light chain and the other half consists of a similar but truncated heavy chain lacking the binding site for the light chain.  
     
     
         23 . An aglycosylated antibody according to  claim 1  in the form of a pharmaceutical composition comprising a physiologically acceptable diluent or carrier.  
     
     
         24 . An aglycosylated antibody according to  claim 1 , for use in therapy.  
     
     
         25 . The use of an aglycosylated antibody according to  claim 1  for the manufacture of a medicament for use in immunosuppression.  
     
     
         26 . The use according to  claim 25 , in which the medicament is for use in the treatment of recipients of a transplant.  
     
     
         27 . A method of treating a patient having cancer or requiring immunosuppression which comprises administering to said patient a therapeutically effective amount of a ligand or an antibody or fragment thereof according to  claim 1.