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.

We claim:  
     
         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 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.  
     
     
         4 . 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, H84 or H108.  
     
     
         5 . An antibody fragment obtainable by enzymatic cleavage of the humanized antibody of any one of claims  1 - 4 .  
     
     
         6 . An Fab or F(ab′)2 fragment of any one of the humanized antibodies of claims  1 - 4 .  
     
     
         7 . The F(ab′)2 fragment of  claim 6 .  
     
     
         8 . The Fab fragment of  claim 6 .  
     
     
         9 . The humanized antibody or fragment of any one of claims  1 - 8 , which is a single chain antibody.  
     
     
         10 . The humanized antibody or fragment of any one of claims  1 - 9  that is an IgG1 immunoglobulin isotype.  
     
     
         11 . The humanized antibody or fragment of any one of claims  1 - 10 , 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.  
     
     
         12 . 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 - 11 , or a fragment thereof.  
     
     
         13 . A polynucleotide sequence, which when expressed in a suitable host cell, yields an antibody of any one of claims  1 - 11 .  
     
     
         14 . 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.  
     
     
         15 . 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.  
     
     
         16 . 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.  
     
     
         17 . 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, H 19, H40, H41, H42, H44, H72, H77, H82 H83, H84 or H108.  
     
     
         18 . An expression vector for expressing the antibody of any one of claims  1 - 11  comprising the polynucleotide sequence of any one of claims  12 - 17 .  
     
     
         19 . A cell transfected with the expression vector of  claim 18 .  
     
     
         20 . A cell transfected with two expression vectors of  claim 18 , 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.  
     
     
         21 . A cell that is capable of expressing the humanized antibody or fragment of any one of claims  1 - 11 .  
     
     
         22 . The cell of any one of claims  19 - 21 , wherein the cell is selected from the group consisting of a myeloma cell and a chinese hamster ovary (CHO) cell.  
     
     
         23 . A pharmaceutical composition comprising the humanized antibody or fragment of any one of claims  1 - 11 , and a pharmaceutically acceptable excipient.  
     
     
         24 . 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 - 11 .  
     
     
         25 . 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 - 11 .  
     
     
         26 . 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 - 11 .  
     
     
         27 . The method of either of claims  25 - 26 , wherein the subject is diagnosed with clinical or pre-clinical Alzheimer's disease or Down's syndrome.  
     
     
         28 . The method of any one of claims  25 - 26 , wherein the subject is not diagnosed with clinical or pre-clinical Alzheimer's disease or Down's syndrome.  
     
     
         29 . Use of the humanized antibody or a fragment thereof according to any one of claims  1 - 11  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.  
     
     
         30 . Use of the humanized antibody or fragment of any one of claims  1 - 11  for the manufacture of a medicament for treating Alzheimer's disease.