Assay method for alzheimer's disease

• US 7,771,722 B2
• Filed: 08/16/2002
• Issued: 08/10/2010
• Est. Priority Date: 08/17/2001
• Status: Active Grant

• Current Assignee: Eli Lilly and Company, Washington University In St Louis
• Original Assignee: Eli Lilly and Company, Washington University In St Louis
• Inventors: Bales, Kelly R., Cummins, David J., Paul, Steven M., Holtzman, David M., DeMattos, Ronald
• Primary Examiner(s): Kolker; Daniel E.
• Application Number:
  US10/486,908
• Publication Number:
  US 20040248197A1
• Time in Patent Office: 2,916 Days
• Field of Search: None
• US Class Current: 424/139.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/41 Glycosylation, sialylation, or fucosylation
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: G01N 2800/2821 Alzheimer
• CPC Class Codes: G01N 33/6896 Neurological disorders, e.g. Alzheimer's disease

Abstract

A diagnostic test for preclinical and clinical Alzheimer'"'"'s disease is based on plasma levels of Aβ₄₀, Aβ₄₂, their ratio, or their rate of entry following administration of antibodies that sequester Aβ. Alterations of any of these parameters from control values identifies preclinical or clinical Alzheimer'"'"'s disease.

Claims

1. A method to diagnose clinical Alzheimer'"'"'s disease in a subject comprising:

(a) administering to said subject an amount of an antibody which specifically binds an epitope contained within positions 13-28 of Aβ and

increases clearance of soluble and bound forms of Aβ

from the central nervous system,

wherein said amount is effective to increase the levels of circulating Aβ

peptides in the blood of said subject when said subject is in a clinical stage of Alzheimer'"'"'s disease;

(b) measuring the level of at least one of Aβ

₄₀ or Aβ

₄₂, or the ratio of Aβ

₄₀/Aβ

₄₂ in the blood of said subject at a time interval after said administering; and

(c) comparing the measured level of Aβ

₄₀ or Aβ

₄₂, or the measured ratio of Aβ

₄₀/Aβ

₄₂ in said subject with a control value of said level or of said ratio, wherein an elevated measured level of Aβ

₄₀ or Aβ

₄₂ or of the ratio of Aβ

₄₀/Aβ

₄₂ in said subject as compared to said control value identifies said subject as in a clinical stage of Alzheimer'"'"'s disease.

2. The method of claim 1, wherein said time interval is less than 1 week.

3. The method of claim 1, wherein said time interval is less than or equal to 24 hours.

4. The method of claim 3, wherein the time interval is less than or equal to 3 hours.

5. The method of claim 1, wherein said administering is by injection of said antibodies.

6. The method of claim 1, wherein the subject is human and the antibody is a humanized antibody or a fragment thereof capable of binding said epitope.

7. The method of claim 6, wherein the humanized antibody or fragment thereof comprises 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.

8. The method of claim 6, wherein the humanized antibody or fragment thereof comprises a light chain of the sequence given by SEQ ID NO:

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

16.

9. The method of claim 6, wherein the humanized antibody or fragment thereof comprises a light chain comprising a variable region of the sequence given by SEQ ID NO:

7 and a heavy chain comprising a variable region of the sequence given by SEQ ID NO;

14.

10. The method of claim 1, wherein said antibody is a fragment capable of binding said epitope.

11. The method of claim 1, wherein the antibody specifically binds to an epitope of Aβ

to which antibody 266 specifically binds.

12. The method of claim 1, wherein the antibody is a single-chain antibody.

13. The method of claim 6, wherein the humanized antibody or fragment thereof comprises:

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

light chain CDR1;

14. A method to diagnose preclinical Alzheimer'"'"'s disease in a subject comprising:

(a) administering to said subject an amount of an antibody which specifically binds an epitope contained within positions 13-28 of Aβ and

increases clearance of soluble and bound forms of Aβ

from the central nervous system,

wherein said amount is effective to increase the levels of circulating Aβ

peptides in the blood of said subject when said subject is in a preclinical stage of Alzheimer'"'"'s disease;

(b) measuring the level of at least one of Aβ

₄₀ or Aβ

₄₂, or the ratio of Aβ

₄₀/Aβ

₄₂ in the blood of said subject at a time interval after said administering; and

(c) comparing the measured level of Aβ

₄₀ or Aβ

₄₂, or the measured ratio of Aβ

₄₀/Aβ

₄₂ in said subject with a control value of said level or of said ratio, wherein an elevated measured level of Aβ

₄₀ or Aβ

₄₂ or of the ratio of Aβ

₄₀/Aβ

₄₂ in said subject as compared to said control value identifies said subject as in a preclinical stage of Alzheimer'"'"'s disease.

15. The method of claim 14, wherein said time interval is less than 1 week.

16. The method of claim 14, wherein said time interval is less than or equal to 24 hours.

17. The method of claim 16, wherein the time interval is less than or equal to 3 hours.

18. The method of claim 14, wherein said administering is by injection of said antibody.

19. The method of claim 14, wherein the subject is human and the antibody is a humanized antibody or a fragment thereof capable of binding said epitope.

20. The method of claim 19, wherein the humanized antibody or fragment thereof comprises 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.

21. The method of claim 19, wherein the humanized antibody or fragment thereof comprises a light chain of the sequence given by SEQ ID NO:

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

16.

22. The method of claim 19, wherein the humanized antibody or fragment thereof comprises a light chain comprising a variable region of the sequence given by SEQ ID NO:

7 and a heavy chain comprising a variable region of the sequence given by SEQ ID NO;

14.

23. The method of claim 14, wherein said antibody is a fragment capable of binding said epitope.

24. The method of claim 14, wherein the antibody specifically binds to an epitope of Aβ

to which antibody 266 specifically binds.

25. The method of claim 14, wherein the antibody is a single-chain antibody.

26. The method of claim 19, wherein the humanized antibody or fragment thereof comprises:

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

light chain CDR1;

27. A method for diagnosing clinical Alzheimer'"'"'s disease in a subject comprising:

(a) administering to said subject an amount of an antibody which specifically binds an epitope contained within positions 13-28 of Aβ and

increases clearance of soluble and bound forms of Aβ

from the central nervous system,

wherein said amount is effective to increase the levels of circulating Aβ

peptides in the blood of said subject when said subject is in a clinical stage of Alzheimer'"'"'s disease;

(b) selecting one or more of the following components for measurement;

(1) the level of circulating Aβ

₄₀,

(2) the level of circulating Aβ

₄₂, or

(3) the ratio of the level of circulating Aβ

₄₀ to the level of circulating Aβ

₄₂

(c) measuring each of said selected one or more components in a blood sample of said subject at a predetermined time interval after said administering to obtain a measured value of each selected component for said subject; and

(d) comparing each measured value with a preselected control value, wherein an elevated measured value as compared to said control value identifies said subject as being in a clinical stage of Alzheimer'"'"'s disease.

28. A method for diagnosing preclinical Alzheimer'"'"'s disease in a subject comprising:

(a) administering to said subject an amount of an antibody which specifically binds an epitope contained within positions 13-28 of Aβ and

increases clearance of soluble and bound forms of Aβ

from the central nervous system,

wherein said amount is effective to increase the levels of circulating Aβ

peptides in the blood of said subject when said subject is in a preclinical stage of Alzheimer'"'"'s disease;

(b) selecting one or more of the following components for measurement;

(1) the level of circulating Aβ

₄₀,

(2) the level of circulating Aβ

₄₂, or

(3) the ratio of the level of circulating Aβ

₄₀ to the level of circulating Aβ

₄₂

(c) measuring each of said selected one or more components in a blood sample of said subject at a predetermined time interval after said administering to obtain a measured value of each selected component for said subject; and

(d) comparing each measured value with a preselected control value, wherein an elevated measured value as compared to said control value identifies said subject as being in a preclinical stage of Alzheimer'"'"'s disease.