Humanized antibodies that sequester Abeta peptide

• US 20040043418A1
• Filed: 08/21/2002
• Published: 03/04/2004
• Est. Priority Date: 02/24/2000
• Status: Active Grant

• Current Assignee: Eli Lilly and Company, Washington University In St Louis
• Original Assignee: Washington University In St Louis
• Inventors: Vasquez, Maximiliano, Tsurushita, Naoya, Paul, Steven M., Holtzman, David M., Bales, Kelly R., DeMattos, Ronald
• Granted Patent: US 7,195,761 B2
• Time in Patent Office: Days
• US Class Current: 435/7.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• 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 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 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Abstract

A method to treat conditions characterized by formation of amyloid plaques both prophylactically and therapeutically is described. The method employs humanized antibodies which sequester soluble Aβ peptide from human biological fluids or which preferably specifically bind an epitope contained within position 13-28 of the amyloid beta peptide Aβ.

Claims

1. A humanized antibody, or fragment thereof, comprising:

a. a light chain comprising three light chain complementarity determining regions (CDRs) having the following amino acid sequences;

light chain CDR1;

1 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

5 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

10 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

15Arg Ser Ser Gln Ser Leu Ile Tyr Ser Asp Gly Asn Ala Tyr Leu His;

or(SEQ ID NO;

1)  

1 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

5 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

10 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

15Arg Ser Ser Gln Ser Leu Val Tyr Ser Asp Gly Asn Ala Tyr Leu His(SEQ ID NO;

15) light chain CDR2;

 

1 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

5Lys Val Ser Asn Arg Phe Ser and,(SEQ ID NO;

2) light chain CDR3;

1 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

5Ser Gln Ser Thr His Val Pro Trp Thr(SEQ ID NO;

3)

and a light chain framework sequence from a humanized immunoglobulin light chain; and

b. a heavy chain comprising three heavy chain CDRs having the following amino acid sequences;

heavy chain CDR1;

 

1 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

5Arg Tyr Ser Met Ser(SEQ ID NO;

4) heavy chain CDR2;

1 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

5 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

10 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

15Gln Ile Asn Ser Val Gly Asn Ser Thr Tyr Tyr Pro Asp Thr Val Lys Gly;

or(SEQ ID NO;

5)  

1 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

5 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

10 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

15Gln IIe Asn Ser Val Gly Asn Ser Thr Tyr Tyr Pro Asp Ser Val Lys Gly and,(SEQ ID NO;

16) heavy chain CDR3;

 

1Gly Asp Tyr(SEQ ID NO;

6)

and a heavy chain framework sequence from a humanized immunoglobulin heavy chain.

2. The humanized antibody, or fragment thereof, of claim 1, wherein light chain CDR1 is:

3. The humanized antibody, or fragment thereof, of claim 1 comprising a humanized light chain variable region comprising the following sequence:

4. The humanized antibody or fragment thereof of claim 1 having a light chain variable region of the sequence given by SEQ ID NO:

9 and a heavy chain variable region given by SEQ ID NO;

10.

5. The humanized antibody or fragment thereof of claim 4 having a light chain of the sequence given by SEQ ID NO:

11 and a heavy chain of the sequence given by SEQ ID NO;

12.

6. The humanized antibody fragment of claim 1.

7. The humanized antibody fragment of claim 5.

8. The humanized antibody or fragment of claim 1 that is an IgG₁ immunoglobulin isotype.

9. The humanized antibody or fragment of claim 1, wherein the antibody or fragment thereof is produced in a host cell selected from the group consisting of a myeloma cell, a chinese hamster ovary cell, a syrian hamster ovary cell, and a human embryonic kidney cell.

10. A polynucleic acid comprising a sequence coding for the light chain or the heavy chain of the humanized antibody of claim 1, or a fragment thereof.

11. The polynucleic acid of claim 10, comprising a sequence coding for the light chain variable region given by SEQ ID NO:

7.

12. The polynucleic acid of claim 10, comprising a sequence coding for the light chain variable region given by SEQ ID NO:

9.

13. The polynucleic acid of claim 10, comprising a sequence coding for the heavy chain variable region given by SEQ ID NO:

8.

14. The polynucleic acid of claim 10, comprising a sequence coding for the heavy chain variable region given by SEQ ID NO:

10.

15. The polynucleic acid of claim 10, comprising a sequence coding for the light chain given by SEQ ID NO:

11.

16. The polynucleic acid of claim 10, comprising a sequence coding for the heavy chain given by SEQ ID NO:

12.

17. A polynucleic acid comprising a sequence coding for the light chain or the heavy chain of the humanized antibody of claim 5, or a fragment thereof.

18. A polynucleic acid, which when expressed in a suitable host cell, yields the antibody or fragment of claim 1.

19. A polynucleic acid, which when expressed in a suitable host cell, yields the antibody or fragment of claim 5.

20. An expression vector for expressing the antibody or fragment of claim 1 comprising nucleotide sequences encoding said antibody or fragment.

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. An expression vector for expressing the antibody or fragment of claim 5 comprising nucleotide sequences encoding said antibody or fragment.

24. A cell transfected with the expression vector of claim 23.

25. A cell transfected with two expression vectors of claim 23, 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.

26. A cell that is capable of expressing the humanized antibody or fragment thereof of claim 1.

27. A cell that is capable of expressing the humanized antibody or fragment thereof of claim 5.

28. A method to inhibit the formation of amyloid plaques or the effects of toxic soluble Aβ

species in humans, which method comprises administering to a human subject in need of such inhibition an effective amount of the humanized antibody or fragment thereof of claim 1.

29. A method to inhibit the formation of amyloid plaques or the effects of toxic soluble Aβ

species in humans, which method comprises administering to a human subject in need of such inhibition an effective amount of the humanized antibody or fragment thereof of claim 5.

30. A method to reduce the formation of amyloid plaques or the effects of toxic soluble Aβ

species in humans, which method comprises administering to a human subject in need of such inhibition an effective amount of the humanized antibody or fragment thereof of claim 1.

31. A method to reduce the formation of amyloid plaques or the effects of toxic soluble Aβ

species in humans, which method comprises administering to a human subject in need of such inhibition an effective amount of the humanized antibody or fragment thereof of claim 5.

32. A method of reversing cognitive decline in a subject, comprising administering to the subject an effective amount of a humanized antibody of claim 1 or a fragment thereof.

33. A method of reversing cognitive decline in a subject, comprising administering to the subject an effective amount of a humanized antibody of claim 5 or a fragment thereof.

34. A method of improving cognition in a subject, comprising administering to the subject an effective amount of a humanized antibody of claim 1 or a fragment thereof.

35. A method of improving cognition in a subject, comprising administering to the subject an effective amount of a humanized antibody of claim 5 or a fragment thereof.

36. A method of treating cognitive decline in a subject, comprising administering to the subject an effective amount of a humanized antibody of claim 1 or a fragment thereof.

37. A method of treating cognitive decline in a subject, comprising administering to the subject an effective amount of a humanized antibody of claim 5 or a fragment thereof.

38. A method of preventing cognitive decline in a subject, comprising administering to the subject an effective amount of a humanized antibody of claim 1 or a fragment thereof.

39. A method of preventing cognitive decline in a subject, comprising administering to the subject an effective amount of a humanized antibody of claim 5 or a fragment thereof.

40. A method of treating, preventing, or reversing cognitive decline in clinical or pre-clinical Alzheimer'"'"'s disease, Down'"'"'s syndrome, or clinical or pre-clinical cerebral amyloid angiopathy comprising:

expressing recombinant sequences of the antibody or fragment thereof of claim 1 in human tissues.

41. A method of treating, preventing, or reversing cognitive decline in clinical or pre-clinical Alzheimer'"'"'s disease, Down'"'"'s syndrome, or clinical or pre-clinical cerebral amyloid angiopathy comprising:

expressing recombinant sequences of the antibody or fragment thereof of claim 5 in human tissues.

42. A method of treating Alzheimer'"'"'s disease, comprising administering to a patient in need thereof an effective amount of the antibody or fragment of claim 1.

43. A method of treating Alzheimer'"'"'s disease, comprising administering to a patient in need thereof an effective amount of the antibody or fragment of claim 5.

44. A pharmaceutical composition that comprises the humanized antibody or fragment of claim 1, and a pharmaceutically acceptable excipient.

45. A pharmaceutical composition that comprises the humanized antibody or fragment of claim 5, and a pharmaceutically acceptable excipient.