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 sequence:  
     
       
         
               
               
             
                   
               
                 (SEQ ID NO: 1) 
                   
               
               
               
             
                 (a) Ser-Phe-Pro-Met-Ala, 
                   
               
                   
               
               
               
             
                 (SEQ ID NO: 2) 
                   
               
               
               
             
                 (b) Thr-Ile-Ser-Thr-Ser-Gly-Gly-Arg-Thr-Tyr-Tyr- 
                   
               
                 Arg-Asp-Ser-VaI-Lys-Gly, 
               
                   
               
               
               
             
                 (SEQ ID NO: 3) 
                   
               
               
               
             
                 (c) Phe-Arg-Gln-Tyr-Ser-Gly-Gly-Phe-Asp-Tyr, 
                   
               
                   
               
               
               
             
                 (SEQ ID NO: 4) 
                   
               
               
               
             
                 (d) Thr-Leu-Ser-Ser-Gly-Asn-IIe-Glu-Asn-Asn-Tyr- 
                   
               
                 Val-His, 
               
                   
               
               
               
             
                 (SEQ ID NO: 5) 
                   
               
               
               
             
                 (e) Asp-Asp-Asp-Lys-Arg-Pro-Asp, 
                   
               
                   
               
               
               
             
                 (SEQ ID NO: 6) 
                   
               
               
               
             
                 (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:  
     
       
         
               
               
             
                   
               
                 (SEQ ID NO: 1) 
                   
               
               
               
             
                 (a) Ser-Phe-Pro-Met-Ala, 
                   
               
                   
               
               
               
             
                 (SEQ ID NO: 2) 
                   
               
               
               
             
                 (b) Thr-Ile-Ser-Thr-Ser-Gly-Gly-Arg-Thr-Tyr-Tyr- 
                   
               
                 Arg-Asp-Ser-Val-Lys-Gly, 
               
                   
               
               
               
             
                 (SEQ ID NO: 3) 
                   
               
               
               
             
                 (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:  
       
         
           
                 
                 
               
                     
                 
                   (SEQ ID NO: 4) 
                     
                 
                 
                 
               
                   (d) Thr-Leu-Ser-Ser-Gly-Asn-Ile Glu-Asn-Asn-Tyr- 
                     
                 
                   Val-His, 
                 
                     
                 
                 
                 
               
                   (SEQ ID NO: 5) 
                     
                 
                 
                 
               
                   (e) Asp-Asp-Asp-Lys-Arg-Pro-Asp, 
                     
                 
                     
                 
                 
                 
               
                   (SEQ ID NO: 6) 
                     
                 
                 
                 
               
                   (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:  
     
       
         
               
               
             
                   
               
                 (SEQ ID NO: 1) 
                   
               
               
               
             
                 (a)Ser-Phe-Pro-met-Ala, 
                   
               
                   
               
               
               
             
                 (SEQ ID NO: 2) 
                   
               
               
               
             
                 (b)Thr-Ile-Ser-Thr-Ser-Gly-Gly-Arg-Thr-Tyr-Tyr- 
                   
               
                 Arg-Asp-Ser-Val-Lys-Gly, 
               
                   
               
               
               
             
                 (SEQ ID NO: 3) 
                   
               
               
               
             
                 (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:  
       
         
           
                 
                 
               
                     
                 
                   (SEQ ID NO: 4) 
                     
                 
                 
                 
               
                   (d) Thr-Leu-Ser-Ser-Gly-Asn-Ile Glu-Asn-Asn-Tyr- 
                     
                 
                   Val-His, 
                 
                     
                 
                 
                 
               
                   (SEQ ID NO: 5) 
                     
                 
                 
                 
               
                   (e) Asp-Asp-Asp-Lys-Arg-Pro-Asp, 
                     
                 
                     
                 
                 
                 
               
                   (SEQ ID NO: 6) 
                     
                 
                 
                 
               
                   (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  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  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-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/CDR/-Trp-Gly-Gln-Gly-Thr- 
               
                 Leu-Val-Thr-Val-Ser-Ser, (SEQ ID NO: 7/CDR/ 
               
                 SEQ ID NO: 8/CDR/SEQ ID NO: 9/CDR/SEQ ID NO: 10), 
               
                   
               
           
              
              
              
              
              
              
              
              
              
              
              
             
          
         
       
     
     ID NO: 8/CDR/SEQ ID NO: 9/CDR/SEQ ID NO: 10), 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- 
               
                 Ile-Phe-/CDR/-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-/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 (SEQ ID NO: 12/ 
               
                 CDR/SEQ ID NO: 13/CDR/SEQ ID NO: 14/CDR/ 
               
                 SEQ ID NO: 26), 
               
                   
               
           
              
              
              
              
              
              
              
              
              
              
              
              
              
             
          
         
       
     
     ID NO: 14/CDR/SEQ ID NO: 26), 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  
     
       
         
               
               
             
                   
               
                 (SEQ ID NO: 11) 
                   
               
               
               
             
                 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-Vat-Thr-Val-Ser-Ser. 
               
                   
               
           
              
              
             
          
           
              
              
              
              
              
              
              
              
              
              
              
             
          
         
       
     
   
   
       12 . An aglycosylated antibody according to  claim 8  having a light chain variable domain which comprises  
     
       
         
               
               
             
                   
               
                 (SEQ ID NO: 25) 
                   
               
               
               
             
                 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 isotype.  
   
   
       18 . An aglycosylated antibody according to  claim 15 , in which asparagine 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 the  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.