Humanized antibodies that recognize beta amyloid peptide

Abstract

The invention provides improved agents and methods for treatment of diseases associated with amyloid deposits of Aβ in the brain of a patient. Preferred agents include humanized antibodies.

1 . Humanized 3D6 antibody.  
     
     
         2 . A humanized antibody, or fragment thereof, comprising a humanized light chain comprising three light chain complementarily determining regions (CDRs) from the mouse monoclonal antibody 3D6 and a light chain variable region framework sequence from a human immunoglobulin light chain; and a humanized heavy chain comprising three heavy chain CDRs from the mouse monoclonal antibody 3D6 and a heavy chain variable region framework sequence from a human immunoglobulin heavy chain; wherein the light chain CDRs have the following amino acid sequences:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   light chain CDR1: 
                     
                 
                     
                   Ser Ser Gln Ser Leu Leu Asp Ser Asp Gly Lys Thr 
                 
                     
                   Tyr Leu Asn 
                 
                     
                   (residues 24-39 of SEQ ID NO:2) 
                 
                     
                     
                 
                     
                   light chain CDR2: 
                 
                     
                   Leu Val Ser Lys Leu Asp Ser 
                 
                     
                   (residues 55-61 of SEQ ID NO:2) 
                 
                     
                     
                 
                     
                   light chain CDR3: 
                 
                     
                   Trp Gln Gly Thr His Phe Pro Arg Thr 
                 
                     
                   (residues 94-102 of SEQ ID NO:2) 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       and the heavy chain CDRs have the following amino acid sequences:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   heavy chain CDR1: 
                     
                 
                     
                   Asn Tyr Gly Met Ser 
                 
                     
                   (residues 31-35 of SEQ ID NO:4) 
                 
                     
                     
                 
                     
                   heavy chain CDR2: 
                 
                     
                   Ser Ile Arg Ser Gly Gly Gly Arg Thr Tyr Tyr Ser 
                 
                     
                   Asp Asn Val Lys Gly 
                 
                     
                   (residues 50-66 of SEQ ID NO:4), 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   heavy chain CDR3: 
                 
                     
                   Tyr Asp His Tyr Ser Gly Ser Ser Asp Tyr 
                 
                     
                   (residues 99-107 of SEQ ID NO:4). 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         3 . A humanized antibody of  claim 1  or  2 , wherein the light chain variable region framework sequence is from a human acceptor immunoglobulin light chain designated Kabat ID 019230.  
     
     
         4 . A humanized antibody of  claim 1  or  2 , wherein the heavy chain variable region framework sequence is from a human acceptor immunoglobulin heavy chain designated Kabat ID 045919.  
     
     
         5 . A humanized antibody or fragment thereof comprising a humanized light chain variable region having complementarily determining regions (CDRs) from SEQ ID NO:2 and variable region framework sequence from a human immunoglobulin light chain having substantial sequence identity to the sequence of SEQ ID NO:6, and a humanized heavy chain variable region having CDRs from SEQ ID NO:4 and variable region framework sequence from a human immunoglobulin heavy chain having substantial sequence identity to the sequence of SEQ ID NO:9.  
     
     
         6 . A humanized antibody or fragment thereof comprising a humanized light chain variable region sequence substantially identical to SEQ ID NO:5 and having a substitution in at least one of positions L7, L10, L12, L15, L17, L39, L45, L63, L78, L83, L85, L100 or L104, and a humanized heavy chain variable region sequence substantially identical to SEQ ID NO:8 and having a substitution in at least one of the positions H3, H5, H13, H16, H19, H40, H41, H42, H44, H72, H77, H82H83, H84 or H108.  
     
     
         7 . An antibody fragment obtainable by enzymatic cleavage of the humanized antibody of any one of claims  1 - 6 .  
     
     
         8 . An Fab or F(ab′)2 fragment of any one of the humanized antibodies of claims  1 - 6 .  
     
     
         9 . The F(ab′)2 fragment of  claim 8 .  
     
     
         10 . The Fab fragment of  claim 8 .  
     
     
         11 . The humanized antibody or fragment of any one of claims  1 - 10 , which is a single chain antibody.  
     
     
         12 . The humanized antibody or fragment of any one of claims  1 - 11  that is an IgG1 immunoglobulin isotype.  
     
     
         13 . The humanized antibody or fragment of any one of claims  1 - 12 , wherein the antibody or fragment thereof is produced in a host cell selected from the group consisting of a myeloma cell and a chinese hamster ovary (CHO) cell.  
     
     
         14 . A polynucleotide compound, comprising a sequence coding for the light chain or the heavy chain of the humanized antibody of any one of claims  1 - 13 , or a fragment thereof.  
     
     
         15 . A polynucleotide sequence, which when expressed in a suitable host cell, yields an antibody of any one of claims  1 - 13 .  
     
     
         16 . A polynucleotide comprising a sequence that codes for a light chain variable region having complementarily determining regions (CDRs) from SEQ ID NO:2 and variable region framework sequence from a human immunoglobulin light chain having substantial sequence identity to the sequence of SEQ ID NO:6.  
     
     
         17 . A polynucleotide comprising a sequence that codes for a light chain variable region sequence substantially identical to SEQ ID NO:5 and having a substitution in at least one of positions L7, L10, L12, L15, L17, L39, L45, L63, L78, L83, L85, L100 or L104.  
     
     
         18 . A polynucleotide comprising a sequence that codes for a heavy chain variable region having CDRs from SEQ ID NO:4 and variable region framework sequence from a human immunoglobulin heavy chain having substantial identity to the sequence of SEQ ID NO:9.  
     
     
         19 . A polynucleotide comprising a sequence that codes for a heavy chain variable region sequence substantially identical to SEQ ID NO:8 an having a substitution in at least one of the positions H3, H5, H13, H16, H19, H40, H41, H42, H44, H72, H77, H82H83, H84 or H108.  
     
     
         20 . An expression vector for expressing the antibody of any one of claims  1 - 11  comprising the polynucleotide sequence of any one of claims  15 - 19 .  
     
     
         21 . A cell transfected with the expression vector of  claim 20 .  
     
     
         22 . A cell transfected with two expression vectors of  claim 20 , wherein a first vector comprises the polynucleotide sequence coding for the light chain and a second vector comprises the sequence coding for the heavy chain.  
     
     
         23 . A cell that is capable of expressing the humanized antibody or fragment of any one of claims  1 - 13 .  
     
     
         24 . The cell of any one of claims  21 - 23 , wherein the cell is selected from the group consisting of a myeloma cell and a chinese hamster ovary (CHO) cell.  
     
     
         25 . A pharmaceutical composition comprising the humanized antibody or fragment of any one of claims  1 - 13 , and a pharmaceutically acceptable excipient.  
     
     
         26 . A method of treating Down's syndrome or clinical or pre-clinical Alzheimer's disease in a human subject, comprising administering to the human subject an effective amount of a humanized antibody or fragment of any one of claims  1 - 13 .  
     
     
         27 . A method to inhibit the formation of Aβ plaque in the brain of a human subject, comprising administering to the human subject an effective amount of the humanized antibody or fragment of any one of claims  1 - 13 .  
     
     
         28 . A method to reduce Aβ plaque in the brain of a human subject, comprising administering to the human subject an effective amount of a humanized antibody or fragment of any one of claims  1 - 13 .  
     
     
         29 . The method of either of claims  27 - 28 , wherein the subject is diagnosed with clinical or pre-clinical Alzheimer's disease or Down's syndrome.  
     
     
         30 . The method of any one of claims  27 - 28 , wherein the subject is not diagnosed with clinical or pre-clinical Alzheimer's disease or Down's syndrome.  
     
     
         31 . Use of the humanized antibody or a fragment thereof according to any one of claims  1 - 13  for the manufacture of a medicament, including prolonged expression of recombinant sequences of the antibody or antibody fragment in human tissues, for treating clinical or pre-clinical Alzheimer's disease or Down's syndrome.  
     
     
         32 . Use of the humanized antibody or fragment of any one of claims  1 - 13  for the manufacture of a medicament for treating Alzheimer's disease.  
     
     
         33 . An antibody comprising a sequence having at least 95% identity with the sequence given by SEQ ID NO: 71 and a sequence having at least 95% identity with the sequence given by SEQ ID NO: 72.  
     
     
         34 . A polynucleotide compound, comprising a sequence coding for either a sequence having at least 95% identity with SEQ ID NO: 71 or a sequence having at least 95% identity with SEQ ID NO: 72 of the antibody of  claim 33 .  
     
     
         35 . A cell that is capable of expressing the antibody of  claim 33 .  
     
     
         36 . A pharmaceutical composition, comprising the antibody of  claim 33  and a pharmaceutically acceptable excipient.  
     
     
         37 . A method for treating clinical or pre-clinical Alzheimer's disease or Down's syndrome in a human subject, comprising administering to the human subject an effective amount of the antibody of  claim 33 .  
     
     
         38 . An antibody comprising a light chain comprising a sequence having at least 95% identity with the sequence given by SEQ ID NO: 71 and a heavy chain comprising a sequence having at least 95% identity with the sequence given by SEQ ID NO: 72.  
     
     
         39 . A polynucleotide compound, comprising a sequence coding for either a sequence having at least 95% identity with SEQ ID NO: 71 or a sequence having at least 95% identity with SEQ ID NO: 72 of the antibody of  claim 38 .  
     
     
         40 . A cell that is capable of expressing the antibody of  claim 38 .  
     
     
         41 . A pharmaceutical composition, comprising the antibody of  claim 38  and a pharmaceutically acceptable carrier.  
     
     
         42 . A method for treating clinical or pre-clinical Alzheimer's disease or Down's syndrome in a human subject, comprising administering to the human subject an effective amount of the antibody of  claim 38 .  
     
     
         43 . The antibody of  claim 38  having a light chain which further comprises light chain constant region sequence from human IgG1 and a heavy chain which further comprises heavy chain constant region sequence from human IgG1.  
     
     
         44 . A polynucleotide compound, comprising a sequence coding either the light chain or heavy chain of the antibody of  claim 43 .  
     
     
         45 . A cell that is capable of expressing the antibody of  claim 43 .  
     
     
         46 . A pharmaceutical composition, comprising the antibody of  claim 43  and a pharmaceutically acceptable excipient.  
     
     
         47 . A method for treating clinical or pre-clinical Alzheimer's Disease or Down's syndrome in a human subject, comprising administering to the human subject an effective amount of the antibody of  claim 43 .  
     
     
         48 . An antibody fragment comprising a sequence having at least 95% identity with the sequence given by SEQ ID NO: 71 and comprising a sequence having at least 95% identity with the sequence given by SEQ ID NO: 72  
     
     
         49 . The antibody fragment of  claim 48 , which is an Fab or an F(ab′)2 fragment.  
     
     
         50 . The antibody fragment of  claim 48 , which is a single chain.  
     
     
         51 . A polynucleotide compound, comprising a sequence coding for either the sequence having at least 95% identity with SEQ ID NO: 71 or the sequence having at least 95% identity with SEQ ID NO: 72 of the antibody fragment of  claim 48 .  
     
     
         52 . A cell that is capable of expressing the antibody fragment of  claim 48 .  
     
     
         53 . A pharmaceutical composition, comprising the antibody fragment of  claim 48  and a pharmaceutically acceptable excipient.  
     
     
         54 . A method for treating clinical or pre-clinical Alzheimer's disease or Down's syndrome in a human subject, comprising administering to the human subject an effective amount of the antibody fragment of  claim 48.