Humanized antibodies that recognize beta amyloid peptide

• US 20080050367A1
• Filed: 09/05/2006
• Published: 02/28/2008
• Est. Priority Date: 04/07/1998
• Status: Abandoned Application

• Current Assignee: Janssen Alzheimer Immunotherapy (Johnson & Johnson), Wyeth LLC (Pfizer Inc.)
• Original Assignee: Wyeth LLC (Pfizer Inc.)
• Inventors: Saldanha, Jose, Basi, Guriq, Schenk, Dale
• Application Number:
  US11/516,724
• Publication Number:
  US 20080050367A1
• Time in Patent Office: Days
• US Class Current: 424/133.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2039/53 DNA (RNA) vaccination
• CPC Class Codes: A61K 2039/55505 Inorganic adjuvants
• CPC Class Codes: A61K 2039/55555 Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers
• CPC Class Codes: A61K 2039/55566 Emulsions, e.g. Freund's adjuvant, MF59
• CPC Class Codes: A61K 2039/55572 Lipopolysaccharides; Lipid A; Monophosphoryl lipid A
• CPC Class Codes: A61K 2039/55577 Saponins; Quil A; QS21; ISCOMS
• CPC Class Codes: A61K 2039/6037 Bacterial toxins, e.g. diphteria toxoid [DT], tetanus toxoid [TT]
• CPC Class Codes: A61K 2039/605 MHC molecules or ligands thereof
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/00 Medicinal preparations containing organic active ingredients
• CPC Class Codes: A61K 31/739 Lipopolysaccharides
• CPC Class Codes: A61K 38/193 Colony stimulating factors [CSF]
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61K 39/0007 Nervous system antigens; Prions
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 47/646 the entire peptide or protein drug conjugate elicits an immune response, e.g. conjugate vaccines
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61K 9/2009 Inorganic compounds
• CPC Class Codes: A61K 9/2031 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, polyethylene oxide, poloxamers
• CPC Class Codes: A61K 9/2054 : Cellulose; Cellulose derivatives, e.g. hydroxypropyl methylcellulose
• CPC Class Codes: A61K 9/4866 : Organic macromolecular compounds
• CPC Class Codes: A61K 9/7023 : Transdermal patches and similar drug-containing composite devices, e.g. cataplasms galenical aspects of iontophoretic devices A61K9/0009; microneedle arrays A61K9/0021; buccal patches A61K9/006
• CPC Class Codes: A61P 25/00 : Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/28 : for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: C07K 14/4711 : Alzheimer's disease; Amyloid plaque core protein
• CPC Class Codes: C07K 16/18 : against material from animals or humans
• CPC Class Codes: C07K 2317/24 : containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/34 : Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/565 : Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/567 : Framework region [FR]
• CPC Class Codes: C07K 2317/77 : Internalization into the cell
• CPC Class Codes: C07K 2317/92 : Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide

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.

Claims

1. Humanized 3D6 antibody.

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.

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.

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.

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;

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.

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.

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.