Anti-CD19 antibody having ADCC and CDC functions and improved glycosylation profile

Abstract

The present invention relates to an anti-CD19 antibody having a variant Fc region having some specific amino acid modifications relative to a wild-type Fc region which confer one or several useful effector functions. The present invention relates in particular to chimeric, humanized or full human anti-CD19 antibodies comprising such a variant Fc region. It relates advantageously to antibodies with an interesting and valuable glycosylation profile, especially a low fucose level and/or a high oligomannose level and low level of sialylated glycoform. The present invention also relates to the use of these antibodies in the treatment, prevention or management of disease or disorder, such as cancer, especially a B-cell malignancy, and auto-immune disease.

The invention claimed is: 
     
       1. An anti-CD19 antibody modified to comprise a variant human IgG IgG1 Fc region comprising an amino acid substitution at each of the amino acid positions 243, 292, 300, 305, 326, 396, and 333 of the human IgG Fc region, wherein the numbering of the amino acid residues in the Fc region is of the Kabat, wherein the modified antibody is ADCC+ and CDC± CDC+, and wherein the modified antibody comprises the CDRs of the antibodies mR005-1 or mR005-2 whose VH and VL amino acid sequences are depicted on the following table: 
       
         
           
                 
                 
                 
                 
               
                     
                     
                 
                     
                     
                   Amino acid sequence VH 
                   Amino acid sequence VL 
                 
                     
                     
                 
                     
                   R005-1 
                   SEQ ID NO: 29 
                   SEQ ID NO: 31 
                 
                     
                   R005-2 
                   SEQ ID NO: 33 
                   SEQ ID NO: 35. 
                 
                     
                     
                 
             
                
                
                
               
               
                
                
                
               
            
           
         
       
     
     
       2. The antibody of  claim 1 , wherein said antibody has been produced in wild-type rodent cells. 
     
     
       3. The antibody of  claim 1 , which has one or two Fc bearing no (GlcNAc) 2 (Fuc), +(Man) 3 (GlcNAc) 2  glycan. 
     
     
       4. The antibody of  claim 1 , which has one or two Fc bearing no (Gal), (GlcNAc) 2 (Fuc), +(Man) 3 (GlcNAc) 2 glycan. 
     
     
       5. The antibody of  claim 1 , comprising one or two Fc bearing a (Man) 5 (GlcNAc) 2  glycan. 
     
     
       6. The antibody of  claim 1 , comprising one or two Fc bearing one or two of the following glycans:
 (Gal), (GlcNAc) 2 (Fuc), (NeuAc), +(Man) 3 (GlcNAc) 2    
 (Gal) 2 (GlcNAc) 2 (Fuc), (NeuAc), +(Man) 3 (GlcNAc) 2 . 
 
     
     
       7. The antibody of  claim 1 , wherein the antibody recognizes a non-internalizing epitope on the CD19 antigen. 
     
     
       8. The antibody of  claim 1 , which comprises an IgG1 Fc region in which Phe243 is substituted by Leu, Arg292 is substituted by Pro, Tyr300 is substituted by Leu, Val305 is substituted by Leu, Lys326 is substituted by Ala, Glu333 is substituted by Ala, and Pro396 is substituted by Leu. 
     
     
       9. The antibody of  claim 1 , wherein said antibody is a chimeric antibody, a humanized antibody, a full human antibody, a bispecific antibody, or an antibody drug conjugateor an antibody fragment. 
     
     
       10. The antibody of  claim 1 , that triggers programmed cell death. 
     
     
       11. A pool of antibodies according to  claim 1 , wherein it comprises less or equal than 15% of such antibodies comprising one or two Fc bearing a (GlcNAc) 2 (Fuc), +(Man) 3 (GlcNAc) 2  glycan and/or less or equal than 20% of such antibodies comprising one or two Fc bearing a (Gal),(GlcNAc) 2 (Fuc), +(Man) 3 (GlcNAc) 2  glycan. 
     
     
       12. The pool of antibodies of  claim 11 , wherein it comprises at least 15, 20, 30, 40 or 50% of antibodies comprising one or two Fc bearing (Man) 5 (GlcNAc) 2  glycans. 
     
     
       13. The pool of antibodies of  claim 11 , wherein it comprises
 less than 1.5 or 1% of antibodies comprising one or two Fc bearing (Gal),(GlcNAc) 2 (Fuc),(NeuAc), +(Man) 3 (GlcNAc) 2      and/or less than 2 or 1.5% of antibodies comprising one or two Fc bearing (Gal) 2 (GlcNAc) 2 (Fuc), (NeuAc), +(Man) 3 (GlcNAc) 2 .   
     
     
       14. A pharmaceutical composition comprising an antibody of  claim 1 , and a physiologically acceptable vehicle or excipient. 
     
     
       15. The antibody of  claim 2 , wherein the wild-type rodent cells are wild-type CHO cells. 
     
     
       16. The antibody of  claim 1 , wherein the IgG Fc region is an IgG1 Fc region. 
     
     
       17. The composition of  claim 14 , further comprising an antibody directed against CD20, CD52, CD22, EGF receptor, VEGF receptor, mimics ganglioside GD3, CEA or HER2. 
     
     
       18. The composition of  claim 14 , comprising further an antibody directed against CD20. 
     
     
       19. The antibody of  claim 8  wherein Glu333 is substituted by Ala. 
     
     
       20. A pharmaceutical composition comprising a monoclonal antibody or a fragment thereof, wherein the monoclonal antibody or fragment thereof specifically binds to CD19 and comprises the CDRs of antibody mR005-1 whose VH and VL amino acid sequences are depicted on the following table: 
       
         
           
                 
                 
                 
                 
               
                     
                     
                 
                     
                     
                   Amino acid sequence VH 
                   Amino acid sequence VL 
                 
                     
                     
                 
                     
                   R005-1 
                   SEQ ID NO: 29 
                   SEQ ID NO: 31 
                 
                     
                     
                 
             
                
                
                
               
               
                
                
               
            
           
         
       
       wherein said monoclonal antibody or fragment thereof triggers programmed cell death; and a physiologically acceptable vehicle or excipient.  
     
     
       21. The pharmaceutical composition according to claim 20, wherein the antibody or fragment thereof comprises:
 a) a VH domain comprising:
 (i) CDR1 of sequence SEQ ID NO: 5; 
 (ii) CDR2 of sequence SEQ ID NO: 6; and 
 (iii) CDR3 of sequence SEQ ID NO: 7; and 
   b) a VL domain comprising:
 (i) CDR1 of sequence SEQ ID NO: 8; 
 (ii) CDR2 of sequence SEQ ID NO: 9; and 
 (iii) CDR3 of sequence SEQ ID NO: 10.  
   
     
     
       22. A pharmaceutical according to claim 20, wherein the antibody or fragment thereof comprises:
 a) a VH domain comprising:
 (i) CDR1 of sequence SEQ ID NO: 11; 
 (ii) CDR2 of sequence SEQ ID NO: 12; and 
 (iii) CDR3 of sequence SEQ ID NO: 13; and 
   b) a VL domain comprising:
 (i) CDR1 of sequence SEQ ID NO: 14; 
 (ii) CDR2 of sequence SEQ ID NO: 15; and 
 (iii) CDR3 of sequence SEQ ID NO: 10.  
   
     
     
       23. A pharmaceutical composition according to claim 20, wherein the antibody or fragment thereof comprises:
 a) a VH domain comprising:
 (i) CDR1 of sequence SEQ ID NO: 16; 
 (ii) CDR2 of sequence SEQ ID NO: 6; and 
 (iii) CDR3 of sequence SEQ ID NO: 13; and 
   b) a VL domain comprising:
 (i) CDR1 of sequence SEQ ID NO: 8; 
 (ii) CDR2 of sequence SEQ ID NO: 9; and 
 (iii) CDR3 of sequence SEQ ID NO: 10.  
   
     
     
       24. The pharmaceutical composition according to claim 20, wherein the antibody further comprises an antibody region selected from the group consisting of human IgG1 Fc, a human kappa region, and a combination thereof.  
     
     
       25. The pharmaceutical composition according to claim 20, wherein said monoclonal antibody or fragment thereof comprises a VH domain of sequence SEQ ID NO: 29 and a VL domain of sequence SEQ ID NO: 31.  
     
     
       26. The pharmaceutical composition of claim 20 further comprising an antibody directed against a tumoral antigen that is different from CD19.  
     
     
       27. The pharmaceutical composition of claim 20 further comprising an antibody directed against CD20.  
     
     
       28. The pharmaceutical composition of claim 20, wherein said monoclonal antibody further comprises a variant human IgG1 Fc region comprising an amino acid substitution at each of the amino acid positions 243, 292, 300, 305, 326, 396, and 333 of the human IgG Fc region, wherein the numbering of the amino acid residues in the Fc region is of the Kabat, wherein the modified antibody is ADCC+ and CDC+.  
     
     
       29. The pharmaceutical composition of claim 28, wherein said antibody has been produced in wild-type rodent cells.  
     
     
       30. The pharmaceutical composition of claim 29, wherein the wild-type rodent cells are wild-type CHO cells.  
     
     
       31. The pharmaceutical composition of claim 28, wherein said monoclonal antibody comprises an IgG1 Fc region in which Phe243 is substituted by Leu, Arg292 is substituted by Pro, Tyr300 is substituted by Leu, Val305 is substituted by Ile, Lys326 is substituted by Ala, Glu333 is substituted by Ala, and Pro396 is substituted by Leu.  
     
     
       32. The pharmaceutical composition of claim 28, wherein said monoclonal antibody comprises an IgG1 Fc region in which Phe243 is substituted by Leu, Arg292 is substituted by Pro, Tyr300 is substituted by Leu, Val305 is substituted by Leu, Lys326 is substituted by Ala, Glu333 is substituted by Ala and Pro396 is substituted by Leu.  
     
     
       33. The pharmaceutical composition of claim 28, further comprising an antibody directed against CD20, CD52, CD22, EGF receptor, VEGF receptor, mimics ganglioside GD3, CEA or HER2.  
     
     
       34. The pharmaceutical composition of claim 28, further comprising an antibody directed against CD20.