Glycogen synthase kinase-3 inhibitors
Novel peptide inhibitors of GSK-3, compositions containing same and uses thereof are disclosed. The novel peptide inhibitors are substrate-competitive inhibitors and have an amino acid sequence designed so as to bind to a defined binding site subunit in GSK-3. Also disclosed are GSK-3 substrate competitive inhibitors which bind to the defined binding site subunit in the enzyme. Also disclosed are mutants of GSK-3 and uses thereof for identifying a putative GSK-3 substrate competitive inhibitor.
1. A peptide having the amino acid sequence I:
[Y n . . . Y 1 ]ZX 1 X 2 X 3 S( p )[W 1 . . . W m] (I)
wherein,
m equals 1 or 2;
n is 3, 4, 5, 6 or 7, such that said peptide consists of 10 to 13 amino acid residues;
S(p) is a phosphorylated serine residue or a phosphorylated threonine residue;
Z is any amino acid residue excepting serine residue or threonine residue;
Y 1 -Yn and W 1 -Wm are each independently any amino acid residue;
each of X 1 and X 2 is independently a hydrophobic amino acid residue selected from the group consisting of glycine, alanine, leucine, isoleucine, valine, proline, phenylalanine, methionine, cysteine and tryptophan; and
X 3 is a proline residue.
2. The peptide of claim 1 , wherein each of X 1 and X 2 is independently selected from the group consisting of a proline residue and an alanine residue.
3. The peptide of claim 1 , wherein X 1 and X 2 are each a proline residue.
4. The peptide of claim 1 , wherein S(p) is a phosphorylated serine.
5. The peptide of claim 1 , wherein Z is an alanine residue.
6. The peptide of claim 1 , wherein m is 1 and W 1 is a proline residue.
7. The peptide of claim 1 , wherein n is 5.
8. The peptide of claim 7 , wherein Y 1 -Y 5 has the amino acid sequence Lys-Glu-Ala-Pro-Pro (SEQ ID NO:48).
9. A peptide having an amino acid sequence selected from the group of amino acid sequences as set forth in SEQ ID NOS:11-13 and 16.
10. The peptide of claim 1 , consisting of the amino acid sequence as set forth in SEQ ID NO:16.
11. The peptide of claim 1 , further comprising a hydrophobic moiety attached thereto.
12. The peptide of claim 11 , wherein said hydrophobic moiety is selected from the group consisting of a fatty acid and a fatty acid attached to an amino acid residue.
13. The peptide of claim 12 , wherein said fatty acid is myristic acid.
14. The peptide of claim 13 , consisting of the amino acid sequence as set forth in SEQ ID NO:17.
15. A pharmaceutical composition comprising, as an active ingredient, the peptide of claim 1 , and a pharmaceutically acceptable carrier.
16. The pharmaceutical composition of claim 15 , being packaged in a packaging material and identified in print, on or in said packaging material, for use in inhibiting an activity of GSK-3.
17. The pharmaceutical composition of claim 15 , being packaged in a packaging material and identified in print, on or in said packaging material, for use in the treatment of a biological condition associated with GSK-3 activity.
18. A method of inhibiting an activity of GSK-3, the method comprising contacting cells expressing GSK-3 with an effective amount of the peptide of claim 1 .
19. A method of treating a biological condition associated with GSK-3 activity, the method comprising administering to a subject in need thereof a therapeutically effective amount of the peptide of claim 1 .
20. The peptide of claim 9 , further comprising a hydrophobic moiety attached thereto.
21. The peptide of claim 20 , wherein said hydrophobic moiety is selected from the group consisting of a fatty acid and a fatty acid attached to an amino acid residue.
22. A peptide having the amino acid sequence I:
[Y n . . . Y 1 ]ZX 1 X 2 X 3 S( p )[W 1 . . . W m] (I)
wherein,
m equals 1 or 2;
n is 3, 4, 5, 6 or 7, such that said peptide consists of 10 to 13 amino acid residues;
S(p) is a phosphorylated serine residue or a phosphorylated threonine residue;
Z is any amino acid residue excepting serine residue or threonine residue;
Y 1 -Yn and W 1 -Wm are each independently any amino acid residue;
X 3 is a hydrophobic amino acid residue selected from the group consisting of glycine, alanine, leucine, isoleucine, valine, proline, phenylalanine, methionine, cysteine and tryptophan; and
X 1 and X 2 are each a proline residue.
23. The peptide of claim 22 , wherein S(p) is a phosphorylated serine.
24. The peptide of claim 22 , wherein Z is an alanine residue.
25. The peptide of claim 22 , wherein m is 1 and W 1 is a proline residue.
26. The peptide of claim 22 , wherein n is 5.
27. The peptide of claim 22 , further comprising a hydrophobic moiety attached thereto.
28. The peptide of claim 27 , wherein said hydrophobic moiety is selected from the group consisting of a fatty acid and a fatty acid attached to an amino acid residue.
29. A pharmaceutical composition comprising, as an active ingredient, the peptide of claim 22 , and a pharmaceutically acceptable carrier.
30. A method of inhibiting an activity of GSK-3, the method comprising contacting cells expressing GSK-3 with an effective amount of the peptide of claim 22 .
31. A method of treating a biological condition associated with GSK-3 activity, the method comprising administering to a subject in need thereof a therapeutically effective amount of the peptide of claim 22 .
32. A peptide having the amino acid sequence I:
[Y n . . . Y 1 ]ZX 1 X 2 X 3 S( p )[W 1 . . . W m] (I)
wherein,
m equals 1 or 2;
n is 5;
S(p) is a phosphorylated serine residue or a phosphorylated threonine residue;
Z is any amino acid residue excepting serine residue or threonine residue;
and W 1 -Wm are each independently any amino acid residue;
each of X 1 , X 2 and X 3 is independently a hydrophobic amino acid residue selected from the group consisting of glycine, alanine, leucine, isoleucine, valine, proline, phenylalanine, methionine, cysteine and tryptophan; and
Y 1 -Y 5 has the amino acid sequence Lys-Glu-Ala-Pro-Pro (SEQ ID NO:48).
33. The peptide of claim 32 , wherein X 3 is selected from the group consisting of a proline residue and an alanine residue.
34. The peptide of claim 32 , wherein S(p) is a phosphorylated serine.
35. The peptide of claim 32 , wherein Z is an alanine residue.
36. The peptide of claim 32 , wherein m is 1 and W 1 is a proline residue.
37. The peptide of claim 32 , further comprising a hydrophobic moiety attached thereto.
38. The peptide of claim 37 , wherein said hydrophobic moiety is selected from the group consisting of a fatty acid and a fatty acid attached to an amino acid residue.
39. A pharmaceutical composition comprising, as an active ingredient, the peptide of claim 32 , and a pharmaceutically acceptable carrier.
40. A method of inhibiting an activity of GSK-3, the method comprising contacting cells expressing GSK-3 with an effective amount of the peptide of claim 32 .
41. A method of treating a biological condition associated with GSK-3 activity, the method comprising administering to a subject in need thereof a therapeutically effective amount of the peptide of claim 32 .
42. A peptide having the amino acid sequence I:
[Y n . . . Y 1 ]ZX 1 X 2 X 3 S( p )[W 1 . . . W m] (I)
wherein,
m equals 1 or 2;
n is 3, 4, 5, 6 or 7, such that said peptide consists of 10 to 13 amino acid residues;
S(p) is a phosphorylated serine residue or a phosphorylated threonine residue;
Z is any amino acid residue excepting serine residue or threonine residue;
Y 1 -Yn and W 1 -Wm are each independently any amino acid residue; and
each of X 1 , X 2 and X 3 is independently a hydrophobic amino acid residue selected from the group consisting of glycine, alanine, leucine, isoleucine, valine, proline, phenylalanine, methionine, cysteine and tryptophan,
the peptide further comprising a hydrophobic moiety attached thereto.
43. The peptide of claim 42 , wherein said hydrophobic moiety is selected from the group consisting of a fatty acid and a fatty acid attached to an amino acid residue.
44. The peptide of claim 43 , wherein said fatty acid is myristic acid.
45. The peptide of claim 42 , wherein X 3 is selected from the group consisting of a proline residue and an alanine residue.
46. The peptide of claim 42 , wherein X 3 is a proline residue.
47. The peptide of claim 42 , wherein each of X 1 and X 2 is independently selected from the group consisting of a proline residue and an alanine residue.
48. The peptide of claim 42 , wherein S(p) is a phosphorylated serine.
49. The peptide of claim 42 , wherein Z is an alanine residue.
50. The peptide of claim 42 , wherein m is 1 and W 1 is a proline residue.
51. The peptide of claim 42 , wherein n is 5.
52. The peptide of claim 51 , wherein Y 1 -Y 5 has the amino acid sequence Lys-Glu-Ala-Pro-Pro (SEQ ID NO:48).
53. A pharmaceutical composition comprising, as an active ingredient, the peptide of claim 42 , and a pharmaceutically acceptable carrier.
54. A method of inhibiting an activity of GSK-3, the method comprising contacting cells expressing GSK-3 with an effective amount of the peptide of claim 42 .
55. A method of treating a biological condition associated with GSK-3 activity, the method comprising administering to a subject in need thereof a therapeutically effective amount of the peptide of claim 42 .