COMPOSITIONS AND METHODS FOR INHIBITING GSK3 ACTIVITY AND USES THEREOF

• US 20090233993A1
• Filed: 03/06/2009
• Published: 09/17/2009
• Est. Priority Date: 03/06/2008
• Status: Abandoned Application

• Current Assignee: Functional Genetics Incorporated, Rockefeller University, Sanford-Burnham Medical Research Institute (Sanford Burnham Prebys Medical Discovery Institute)
• Original Assignee: Burnham Institute For Medical Research
• Inventors: Xu, Huaxi, Li, Wu-bo, Liao, Francesca-Fang, Zhang, Yun-wu, Greengard, Paul, Li, Limin, Cohen, Stanley N.
• Application Number:
  US12/399,850
• Publication Number:
  US 20090233993A1
• Time in Patent Office: Days
• US Class Current: 514/44.R
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• 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 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: G01N 2333/91205 Phosphotransferases in general
• CPC Class Codes: G01N 2800/2821 Alzheimer

Abstract

Disclosed herein are compositions and methods relating to peptides. The peptides can inhibit amyloid beta (Aβ) generation and reduce GSK-3 activities. Further provided are compositions and methods for treating or preventing, for example, Alzheimer'"'"'s disease, cancer, and diabetes.

Claims

1. An isolated nucleic acid comprising a sequence at least 95% identical to SEQ ID NO:

1 or a fragment thereof at least 24 residues in length, wherein the sequence encodes a polypeptide that binds adenylyl cyclase.

2. The isolated nucleic acid of claim 1, wherein the sequence encodes a polypeptide comprising the amino acid sequence SEQ ID NO:

2, or a fragment thereof at least 8 residues in length.

3. The isolated nucleic acid of claim 2, wherein the amino acid sequence comprises amino acids 96 to 141 of SEQ ID NO:

2.

4. The isolated nucleic acid of claim 1, wherein the sequence comprises SEQ ID NO:

1, or a fragment thereof at least 24 residues in length.

5. The isolated nucleic acid of claim 1, wherein the nucleic acid hybridizes under stringent conditions to a hybridization probe consisting of the sequence SEQ ID NO:

1 or the complement of SEQ ID NO;

1.

6. An expression vector comprising the isolated nucleic acid of claim 1, operably linked to an expression control sequence.

7. The expression vector of claim 6, wherein the expression control sequence is a tissue specific promoter.

8. The expression vector of claim 6, wherein the expression control sequence is an inducible promoter.

9. A method comprising administering the expression vector of claim 6 to a subject.

10. A cultured cell, comprising the nucleic acid of claim 1 operably linked to an expression control sequence.

11. The cultured cell of claim 10, wherein the cell is a eukaryotic cell.

12. A method of making a polypeptide, the method comprising culturing the cell of claim 10 under conditions permitting translation of the isolated nucleic acid.

13. A purified polypeptide, comprising an amino acid sequence at least 95% identical to the sequence SEQ ID NO:

2, or a fragment thereof at least 8 residues in length, wherein the polypeptide binds adenylyl cyclase.

14. The purified polypeptide of claim 13, wherein the amino acid sequence comprises the sequence SEQ ID NO:

2, or a fragment thereof at least 8 residues in length.

15. The purified polypeptide of claim 13, wherein the amino acid sequence comprises at least 8 consecutive residues of SEQ ID NO:

2.

16. A method of inhibiting GSK3 activity in a cell, comprising contacting the cell with an isolated nucleic acid comprising a sequence at least 95% identical to SEQ ID NO:

1 or a fragment thereof at least 24 residues in length, operably linked to an expression control sequence, wherein the sequence encodes a polypeptide that binds adenylyl cyclase.

17. The method of claim 16, wherein the isolated nucleic acid encodes a polypeptide comprising the amino acid sequence SEQ ID NO:

2, or a fragment thereof at least 8 residues in length.

18. The method of claim 17, wherein the amino acid sequence comprises amino acids 96 to 141 of SEQ ID NO:

2.

19. The method of claim 16, wherein the isolated nucleic acid comprises SEQ ID NO:

1, or a fragment thereof at least 24 residues in length.

20. The method of claim 16, wherein the isolated nucleic acid hybridizes under stringent conditions to a hybridization probe consisting of the sequence SEQ ID NO:

1 or the complement of SEQ ID NO;

1.

21. The method of claim 16, wherein the cell is in a subject, wherein the cell is contacted with the isolated nucleic acid by administering the isolated nucleic acid to the subject.

22. The method of claim 21, wherein the subject is diagnosed with, suspected of being afflicted with, or identified as being at risk of Alzheimer'"'"'s disease, cancer, diabetes, or a combination.

23. The method of claim 21, wherein the subject is diagnosed with, suspected of being afflicted with, or identified as being at risk of Alzheimer'"'"'s disease.

24. The method of claim 21, wherein the subject is diagnosed with, suspected of being afflicted with, or identified as being at risk of cancer.

25. The method of claim 21, wherein the subject is diagnosed with, suspected of being afflicted with, or identified as being at risk of diabetes.

26. The method of claim 21, wherein the subject is diagnosed with Alzheimer'"'"'s disease, cancer, diabetes, or a combination.

27. The method of claim 21, wherein the subject is diagnosed with Alzheimer'"'"'s disease.

28. The method of claim 21, wherein the subject is diagnosed with cancer.

29. The method of claim 21, wherein the subject is diagnosed with diabetes.

30. The method of claim 21, wherein the subject is suspected of being afflicted with or is identified as being at risk of Alzheimer'"'"'s disease.

31. The method of claim 21, wherein amyloid beta (Aβ

) generation is reduced in the subject.

32. A method of inhibiting GSK3 activity in a cell, comprising contacting the cell with a purified polypeptide comprising an amino acid sequence at least 95% identical to the sequence SEQ ID NO:

2, or a fragment thereof at least 8 residues in length, wherein the polypeptide binds adenylyl cyclase.

33. The method of claim 32, wherein the amino acid sequence comprises amino acids 96 to 141 of SEQ ID NO:

2.

34. The method of claim 32, wherein the cell is in a subject, wherein the cell is contacted with the purified polypeptide by administering the isolated nucleic acid to the subject.

35. The method of claim 34, wherein the subject is diagnosed with, suspected of being afflicted with, or identified as being at risk of Alzheimer'"'"'s disease, cancer, diabetes, or a combination.

36. The method of claim 34, wherein the subject is diagnosed with, suspected of being afflicted with, or identified as being at risk of Alzheimer'"'"'s disease.

37. The method of claim 34, wherein the subject is diagnosed with, suspected of being afflicted with, or identified as being at risk of cancer.

38. The method of claim 34, wherein the subject is diagnosed with, suspected of being afflicted with, or identified as being at risk of diabetes.

39. The method of claim 34, wherein the subject is diagnosed with Alzheimer'"'"'s disease, cancer, diabetes, or a combination.

40. The method of claim 34, wherein the subject is diagnosed with Alzheimer'"'"'s disease.

41. The method of claim 34, wherein the subject is diagnosed with cancer.

42. The method of claim 34, wherein the subject is diagnosed with diabetes.

43. The method of claim 34, wherein the subject is suspected of being afflicted with or is identified as being at risk of Alzheimer'"'"'s disease.

44. The method of claim 34, wherein amyloid beta (Aβ

) generation is reduced in the subject.

45. A method of treating a subject with Alzheimer'"'"'s disease, cancer, or diabetes, the method comprising administering to the subject an expression vector encoding protein FG01, such that a therapeutically effective amount of protein FG01 is expressed in the subject.

46. A method of treating a subject with Alzheimer'"'"'s disease, cancer, or diabetes, the method comprising administering to the subject a cell comprising an expression vector encoding protein FG01, such that a therapeutically effective amount of protein FG01 is expressed by the cell in the subject.

47. The method of claim 46, wherein the cell is a stem cell or progenitor cell.

48. The method of claim 46, wherein the cell is a cell from the subject.

49. A method comprising:

(a) contacting a purified polypeptide comprising an amino acid sequence at least 95% identical to the sequence SEQ ID NO;

2, or a fragment thereof at least 8 residues in length, wherein the polypeptide binds adenylyl cyclase, with a test compound; and

(b) determining whether the test compound binds to the purified polypeptide, said binding being an indication that the test compound is a modulator of glycogen synthase kinase-3 (GSK-3).

50. The method of claim 49 further comprising, following determining whether the test compound binds to the purified polypeptide, making the test compound.

51. The method of claim 49 further comprising, following determining whether the test compound binds to the purified polypeptide, administering the test compound to a subject.

55. A process for making a modulator of glycogen synthase kinase-3 (GSK-3), the method comprising manufacturing the compound identified in claim 49.

52. A method of identifying a modulator of glycogen synthase kinase-3 (GSK-3), the method comprising:

(a) providing a cell comprising a nucleic acid encoding FG01 operably linked to an expression control sequence, wherein the nucleic acid comprises a sequence at least 95% identical to SEQ ID NO;

1, or a fragment thereof at least 24 residues in length, wherein the sequence encodes a polypeptide that binds adenylyl cyclase,

(b) contacting the cell with a test compound; and

(c) measuring expression of FG01, an increase or decrease in FG01 expression an indication that the compound is a modulator of GSK-3.

53. The method of claim 52 further comprising, following measuring expression of FG01, making the test compound.

54. The method of claim 52 further comprising, following measuring expression of FG01, administering the test compound to a subject.

56. A process for making a modulator of glycogen synthase kinase-3 (GSK-3), the method comprising manufacturing the compound identified in claim 52.

57. A method of identifying a compound that binds to FG01, the method comprising:

(a) providing a cell expressing FG01;

(b) contacting the cell with a test compound; and

(c) determining whether the test compound binds to FG01.