Humanized antibodies that recognize beta amyloid peptide

• US 20070154480A1
• Filed: 09/12/2006
• Published: 07/05/2007
• Est. Priority Date: 04/07/1998
• Status: Active Grant

• Current Assignee: Janssen Alzheimer Immunotherapy (Johnson & Johnson), Wyeth LLC (Pfizer Inc.)
• Original Assignee: Wyeth LLC (Pfizer Inc.)
• Inventors: Schenk, Dale
• Granted Patent: US 7,790,856 B2
• Time in Patent Office: Days
• US Class Current: 424/145.100
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: C07K 16/18 against material from animals or humans
• 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 2317/56 variable (Fv) region, i.e. VH and/or VL
• 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 2318/10 Immunoglobulin or domain(s) thereof as scaffolds for inserted non-Ig peptide sequences, e.g. for vaccination purposes

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 10D5 antibody.

3. An antibody fragment obtainable by enzymatic cleavage of the humanized antibody of any one of claims 1-2.

4. An Fab or F(ab′

)2 fragment of any one of the humanized antibodies of claims 1-2.

5. The F(ab′

)2 fragment of claim 4.

6. The Fab fragment of claim 4.

7. The humanized antibody or fragment of any one of claims 1-6, which is a single chain antibody.

8. The humanized antibody or fragment of any one of claims 1-7 that is an IgG1 immunoglobulin isotype.

9. The humanized antibody or fragment of any one of claims 1-8, 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.

10. A polynucleotide compound, comprising a sequence coding for the light chain or the heavy chain of the humanized antibody of any one of claims 1-9, or a fragment thereof.

11. A polynucleotide sequence, which when expressed in a suitable host cell, yields an antibody of any one of claims 1-9.

12. An expression vector for expressing the antibody of any one of claims 1-9 comprising the polynucleotide sequence of any one of claims 10-11.

13. A cell transfected with the expression vector of claim 12.

14. A cell transfected with two expression vectors of claim 12, 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.

15. A cell that is capable of expressing the humanized antibody or fragment of any one of claims 1-9.

16. The cell of any one of claims 13-15, wherein the cell is selected from the group consisting of a myeloma cell and a chinese hamster ovary (CHO) cell.

17. A pharmaceutical composition comprising the humanized antibody or fragment of any one of claims 1-9, and a pharmaceutically acceptable excipient.

18. 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 1-9.

19. 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 1-9.

20. 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 1-9.

21. The method of either of claims 19-20, wherein the subject is diagnosed with clinical or pre-clinical Alzheimer'"'"'s disease or Down'"'"'s syndrome.

22. The method of any one of claims 19-20, wherein the subject is not diagnosed with clinical or pre-clinical Alzheimer'"'"'s disease or Down'"'"'s syndrome.

23. Use of the humanized antibody or a fragment thereof according to any one of claims 1-9 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.

24. Use of the humanized antibody or fragment of any one of claims 1-9 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 10D5 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 10D5 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;

25. At least one isolated mammalian amyloid antibody, comprising either

(i) at least two of the heavy chain complementarity determining regions (CDR) amino acid sequences of at least one of SEQ ID NOS:

67-68;

or

(ii) at least two of the light chain CDR amino acids sequences of at least one of SEQ ID NOS;

65.

30. An amyloid antibody according to any of claims 25-27, wherein said antibody binds amyloid with an affinity of at least one selected from at least 10-9 M and at least 10-10 M.

31. An amyloid antibody according to any of claims 25-27, wherein said antibody substantially modulates at least one activity of at least one amyloid polypeptide.

32. An isolated nucleic acid encoding at least one isolated mammalian amyloid antibody according to any of claims 25-27 and having at least one human CDR of SEQ ID NOS:

15 and 13.

33. An isolated nucleic acid vector comprising an isolated nucleic acid according to claim 32.

34. A prokaryotic or eukaryotic host cell comprising an isolated nucleic acid according to claim 32.

35. A host cell according to claim 34, wherein said host cell is at least one selected from COS-1, COS-7, HEK293, CHO, HeLa or myeloma, cells, or any derivative, immortalized or transformed cell thereof.

36. A method for producing at least one amyloid antibody, comprising translating a nucleic acid according to claim 32 under conditions in vitro, in vivo or in situ, such that the amyloid antibody is expressed in detectable or recoverable amounts.

37. A composition comprising at least one isolated mammalian amyloid antibody according to any of claims 25-27 having at least one human CDR, wherein said antibody specifically binds at least one epitope comprising at least 1-3, to the entire amino acid sequence of SEQ ID NO:

70, and at least one pharmaceutically acceptable carrier or diluent.

38. A composition according to claim 37, further comprising at least one at least one compound or polypeptide selected from at least one of a detectable label or reporter, a drug, an immunomodulation agent, an opthalmic, otic or nasal drug, a topical drug, a nutritional drug, a cytokine, and a cytokine antagonist.

39. An anti-idiotype antibody or fragment that specifically binds at least one amyloid antibody according to any of claim 25-27.

40. A method for diagnosing or treating an amyloid related condition in a cell, tissue, organ or animal, comprising (a) contacting or administering a composition comprising an effective amount of at least one antibody according to any of claim 25-27, with, or to, said cell, tissue, organ or animal.

41. A method according to claim 40, wherein said effective amount is 0.001-50 mg/kilogram of said cells, tissue, organ or animal.

42. A method according to claim 40, wherein said contacting or said administrating is by at least one mode selected from parenteral, subcutaneous, intramuscular, intravenous, intraperitoneal, rectal, intranasal, intrachranial, transdermal, and pulmonary.

43. A method according to claim 40, further comprising administering, prior, concurrently or after said (a) contacting or administering, at least one composition comprising an effective amount of at least one compound or polypeptide selected from at least one of a detectable label or reporter, a drug, an immunomodulation agent, an ophthalmic, otic or nasal drug, a topical drug, a nutritional drug, a cytokine, and a cytokine antagonist.

44. A medical device, comprising at least one amyloid antibody according to any of claim 25-27, wherein said device is suitable to contacting or administering said at least one amyloid antibody by at least one mode selected from parenteral, subcutaneous, intramuscular, intravenous, intracranial, intraperitoneal, bolus, rectal, intranasal, and transdermal.

45. A kit for human pharmaceutical or diagnostic use, comprising packaging material and a container comprising at least one amyloid antibody according to any of claim 25-27.

46. The kit of claim 45, wherein said container is a component of a parenteral, subcutaneous, intramuscular, intravenous, intracranial, intraperitoneal, bolus, rectal, intranasal, or transdermal delivery device or system.

47. A method for producing at least one isolated mammalian amyloid antibody according to any of claim 25-27, comprising providing a host cell or transgenic animal or eukaryotic cell capable of expressing in recoverable amounts said antibody.

48. At least one amyloid antibody produced by a method according to claim 47.

26. At least one isolated mammalian amyloid antibody, comprising at least one heavy chain or light chain CDR having the amino acid sequence of at least one of SEQ ID NOS:

67-68 and 65.

27. At least one isolated mammalian amyloid antibody that binds to the same region of an amyloid polypeptide as an antibody comprising at least one heavy chain or light chain CDR having the amino acid sequence of at least one of SEQ ID NOS:

67-68 and 65.

28. At least one isolated mammalian amyloid antibody, comprising at least one human CDR, wherein said antibody specifically binds at least one epitope selected from amino acids 2-7, 3-8, 33-42, or 34-40 of SEQ ID NO:

70.

29. At least one isolated mammalian amyloid antibody, comprising at least one human CDR, wherein said antibody specifically binds at least one epitope comprising at least 1-3, to the entire amino acid sequence of SEQ ID NO:

70.