Method for enhancing mutant enzyme activities in lysosomal storage disorders

• US 6,589,964 B2
• Filed: 06/14/2002
• Issued: 07/08/2003
• Est. Priority Date: 06/01/1998
• Status: Expired due to Term

• Current Assignee: Icahn School of Medicine at Mount Sinai
• Original Assignee: Icahn School of Medicine at Mount Sinai
• Inventors: Fan, Jian-Qiang, Ishii, Satoshi
• Primary Examiner(s): Barts, Samuel
• Assistant Examiner(s): Khare, Devesh
• Application Number:
  US10/172,604
• Publication Number:
  US 20020198225A1
• Time in Patent Office: 389 Days
• Field of Search: 514/315, 435/208, 536/17.2, 536/4.1
• US Class Current: 514/315
• CPC Class Codes: A61K 31/445 Non condensed piperidines, e.g. piperocaine
• CPC Class Codes: A61K 31/70 Carbohydrates; Sugars; Derivatives thereof sorbitol A61K31/047
• CPC Class Codes: A61K 31/7008 Compounds having an amino group directly attached to a carbon atom of the saccharide radical, e.g. D-galactosamine, ranimustine
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07D 451/06 Oxygen atoms
• CPC Class Codes: C12N 9/2465 acting on alpha-galactose-glycoside bonds, e.g. alpha-galactosidase (3.2.1.22)

Abstract

Method for enhancing in a mammalian cell the activity of an enzyme associated with a lysosomal storage disorder by administering a competitive inhibitor of the enzyme in an amount effective to enhance the activity of the enzyme. Preferred compounds for use in the method are imino sugars and related compounds.

Claims

1. A method of enhancing in a mammalian cell the activity of an enzyme, which method comprises administering a competitive inhibitor of the enzyme in an amount effective to enhance enzyme activity, wherein said competitive inhibitor is a specific competitive inhibitor for said enzyme and dissociates from the enzyme.

2. The method of claim 1, wherein the competitive inhibitor increases the amount of enzyme.

3. The method of claim 1, wherein the mammalian cell comprises a construct that encodes the enzyme, whereby the mammalian cell expresses the enzyme.

4. The method of claim 1, wherein the enzyme is involved in hydrolysis of glycosphingolipids.

5. The method of claim 4, wherein the glycosphingolipid is ceramide trihexoside.

6. The method of claim 1, wherein the enzyme is associated with a glycosphingolipid storage disease.

7. The method of claim 6, wherein the glycosphingolipid storage disease is Fabry disease.

8. The method of claim 6 wherein the enzyme is α

-Gal A.

9. The method of claim 8, wherein the competitive inhibitor is selected from the group consisting of 1-deoxygalactonojirimycin, α

-allo-homonojirimycin, α

-galacto-homonojirimycin, α

-1-C-butyl-deoxynojirimycin, calystegines A₃, calystegine B₂, N-methyl-calystegine A₃ and N-methyl-calystegine B₂, and the enzyme is α

-galactosidase A.

10. The method of claim 9, wherein the competitive inhibitor is 1-deoxygalactonojirimycin.

11. The method of claim 6 wherein the glycosphingolipid storage disease is Pompe disease and the enzyme is α

-glucosidase.

12. The method of claim 11, wherein the competitive inhibitor is selected from the group consisting of 1-deoxynojirimycin, a-homonojirimycin, and castanospermine.

13. The method of claim 6 wherein the glycosphingolipid storage disease is Hurler-Scheie disease and the enzyme is α

-L-iduronidase.

14. The method of claim 13, wherein the competitive inhibitor is selected from the group consisting of 1-deoxyiduronojirimycin and 2-carboxy-3,4,5-trideoxypiperidine.

15. The method of claim 6 wherein the glycosphingolipid storage disease is Sanfilippo disease and the enzyme is α

-N-acetylglucosaminidase.

16. The method of claim 15, wherein the competitive inhibitor is 1,2-dideoxy-2-N-acetamido-nojirimycin.

17. The method of claim 6 wherein the glycosphingolipid storage disease is Fucosidosis and the enzyme is α

-L-fucosidase.

18. The method of claim 17 wherein the competitive inhibitor is selected from the group consisting of 1-deoxyfuconojirimycin, β

-homofuconojirimycin, 2,5-imino-1,2,5-trideoxy-L-glucitol, 2,5-dideoxy-2,5-imino-D-fucitol, and 2,5-imino-1,2,5-trideoxy-D-altritol.

19. The method of claim 6 wherein the glycosphingolipid storage disease is Sialidosis and the enzyme is selected from the group consisting of α

-N-acetyl-neuramimidase and sialidase.

20. The method of claim 19, wherein the competitive inhibitor is selected from the group consisting of 2,5-dideoxy-2,6-imino-sialic acid and Siastatin B.

21. The method of claim 6 wherein the glycosphingolipid storage disease is Schindler-Kanzaki disease and the enzyme is α

-N-acetylgalactosamimidase.

22. The method of claim 21, wherein the competitive inhibitor is 1,2-dideoxy-2-N-acetamido-galactonojirimycin.

23. The method of claim 6 wherein the glycosphingolipid storage disease is GM1-gangliosidosis and the enzyme is acid β

-galactosidase.

24. The method of claim 23, wherein the competitive inhibitor is selected from the group consisting of 4-epi-isofagomine and 1-deoxygalactonojirimycin.

25. The method of claim 6 wherein the glycosphingolipid storage disease is Krabbe disease and the enzyme is galactocerebrosidase.

26. The method of claim 25, wherein the competitive inhibitor is selected from the group consisting of 4-epi-isofagomine and 1-deoxygalactonojirimycin.

27. The method of claim 6 wherein the glycosphingolipid storage disease is Morquio disease and the enzyme is acid β

-galactosidase.

28. The method of claim 27, wherein the competitive inhibitor is selected from the group consisting of 4-epi-isofagomine and 1-deoxygalactonojirimycin.

29. The method of claim 6 wherein the glycosphingolipid storage disease is Tay-Sachs disease and the enzyme is β

-hexosamimidase A.

30. The method of claim 29, wherein the competitive inhibitor is selected from the group consisting of 2-N-acetylamino-isofagomine, 1,2-dideoxy-2-acetamido-nojirimycin, and nagstain.

31. The method of claim 6 wherein the glycosphingolipid storage disease is Sandhoff disease and the enzyme is β

-hexosamimidase B.

32. The method of claim 31, wherein the competitive inhibitor is selected from the group consisting of 2-N-acetamido-isofagomine, 1,2-dideoxy-2-acetamido-nojirimycin, and nagstain.

33. The method of claim 6 wherein the glycosphingolipid storage disease is Sly disease and the enzyme is β

-glucuronidase.

34. The method of claim 33 wherein the competitive inhibitor is selected from the group consisting of 6-carboxy-isofagomine and 2-carboxy-3,4,5-trideoxypiperidine.

35. The method of claim 6 wherein the glycosphingolipid storage disease is β

-mannosidosis and the enzyme is acid β

-mannosidase.

36. The method of claim 35, wherein the competitive inhibitor is 2-hydroxy-isofagomine.

37. The method of claim 6 wherein the glycosphingolipid storage disease is α

-mannosidosis and the enzyme is acid α

-mannosidase.

38. The method of claim 37, wherein the competitive inhibitor is selected from the group consisting of 1-deoxymannojirimycin, swainsonine and mannostatin A.

39. The method of claim 1 wherein the competitive inhibitor comprises an alkaloidal compound.

40. The method of claim 1, wherein the mammalian cell is a human cell.

41. A method of preventing degradation of an enzyme in a mammalian cell, which method comprises administering a competitive inhibitor of the enzyme in an amount effective to enhance enzyme activity, wherein said competitive inhibitor is a specific competitive inhibitor for said enzyme and dissociates from the enzyme.

42. The method of claim 41, wherein the enzyme is a lysosomal enzyme, and wherein said competitive inhibitor binds to the enzyme at neutral pH, but not at lysosomal conditions.

43. The method of claim 42, wherein the neutral pH is around 7.0 and the lysosomal conditionsare at a pH of less than 5.0.

44. The method of claim 42, wherein the enzyme is associated with a glycosphingolipid storage disease.

45. The method of claim 44, wherein the glycosphingolipid storage disease is Fabry disease.

46. The method of claim 45 wherein the enzyme is α

-Gal A.

47. The method of claim 46, wherein the competitive inhibitor is selected from the group consisting of 1-deoxygalactonojirimycin, α

-allo-homonojirimycin, α

-galacto-homonojirimycin, β

-1-C-butyl-deoxynojirimycin, calystegines A₃, calystegine B₂, N-methyl-calystegine A₃ and N-methyl-calystegine B₂.

48. The method of claim 44 wherein the glycosphingolipid storage disease is Pompe disease and the enzyme is α

-glucosidase.

49. The method of claim 48, wherein the competitive inhibitor is selected from the group consisting of 1-deoxynojirimycin, α

-homonojirimycin, and castanospermine.

50. The method of claim 44 wherein the glycosphingolipid storage disease is Hurler-Scheie disease and the enzyme is α

-L-iduronidase.

51. The method of claim 50, wherein the competitive inhibitor is selected from the group consisting of 1-deoxyiduronojirimycin and 2-carboxy-3,4,5-trideoxypiperidine.

52. The method of claim 44 wherein the glycosphingolipid storage disease is Sanfilippo disease and the enzyme is a-N-acetylglucosamimidase.

53. The method of claim 52, wherein the competitive inhibitor is 1,2-dideoxy-2-N-acetamido-nojirimycin.

54. The method of claim 44 wherein the glycosphingolipid storage disease is Fucosidosis and the enzyme is α

-L-fucosidase.

55. The method of claim 54 wherein the competitive inhibitor is selected from the group consisting of 1-deoxyfuconojirimycin, β

-homofuconojirimycin, 2,5-imino-1,2,5-trideoxy-L-glucitol, 2,5-dideoxy-2,5-imino-D-fucitol, and 2,5-imino-1,2,5-trideoxy-D-altritol.

56. The method of claim 44 wherein the glycosphingolipid storage disease is Sialidosis and the enzyme is selected from the group consisting of a-N-acetyl-neuramimidase and sialidase.

57. The method of claim 56, wherein the competitive inhibitor is selected from the group consisting of 2,5-dideoxy-2,6-imino-sialic acid and Siastatin B.

58. The method of claim 44 wherein the glycosphingolipid storage disease is Scindeler-Kanzaki disease and the enzyme is α

-N-acetylgalactosamimidase.

59. The method of claim 58, wherein the competitive inhibitor is 1,2-dideoxy-2-N-acetamido-galactonojirimycin.

60. The method of claim 44 wherein the glycosphingolipid storage disease is GM1-gangliosidosis and the enzyme is acid β

-galactosidase.

61. The method of claim 60, wherein the competitive inhibitor is selected from the group consisting of 4-epi-isofagomine and 1-deoxygalactonojirimycin.

62. The method of claim 44 wherein the glycosphingolipid storage disease is Krabbe disease and the enzyme is galactocerebrosidase.

63. The method of claim 62, wherein the competitive inhibitor is selected from the group consisting of 4-epi-isofagomine and 1-deoxygalactonojirimycin.

64. The method of claim 44 wherein the glycosphingolipid storage disease is Morquio disease and the enzyme is acid β

-galactosidase.

65. The method of claim 64, wherein the competitive inhibitor is selected from the group consisting of 4-epi-isofagomine and 1-deoxygalactonojirimycin.

66. The method of claim 44 wherein the glycosphingolipid storage disease is Tay-Sachs disease and the enzyme is β

-hexosamimidase A.

67. The method of claim 66, wherein the competitive inhibitor is selected from the group consisting of 2-N-acetylamino-isofagomine, 1,2-dideoxy-2-acetamido-nojirimycin, and nagstain.

68. The method of claim 44 wherein the glycosphingolipid storage disease is Sandhoff disease and the enzyme is β

-hexosamimidase B.

69. The method of claim 68, wherein the competitive inhibitor is selected from the group consisting of 2-N-acetamido-isofagomine, 1,2-dideoxy-2-acetamido-nojirimycin, and nagstain.

70. The method of claim 44 wherein the glycosphingolipid storage disease is Sly disease and the enzyme is β

-glucuronidase.

71. The method of claim 70, wherein the competitive inhibitor is selected from the group consisting of 6-carboxy-isofagomine and 2-carboxy-3,4,5-trideoxypiperidine.

72. The method of claim 44 wherein the glycosphingolipid storage disease is β

-mannosidosis and the enzyme is acid β

-mannosidase.

73. The method of claim 72, wherein the competitive inhibitor is 2-hydroxy-isofagomine.

74. The method of claim 44 wherein the glycosphingolipid storage disease is α

-mannosidosis and the enzyme is acid α

-mannosidase.

75. The method of claim 74, wherein the competitive inhibitor is selected from the group consisting of 1-deoxymannojirimycin, swainsonine and mannostatin A.

76. The method of claim 41 wherein the enzyme is involved in hydrolysis of glycosphingolipids.

77. The method of claim 76, wherein the glycosphingolipid is ceramide trihexoside.

78. The method of claim 41 wherein the competitive inhibitor comprises an alkaloidal compound.

79. The method of claim 41, wherein the mammalian cell is transfected to express the enzyme.

80. The method of claim 41, wherein the mammalian cell is a human cell.

81. A method of identifying a compound useful for enhancing the in vivo activity of an enzyme, which method comprises detecting an increase in enzyme activity in the presence of a compound having the following formula:

wherein

R₁ and R₁′

independently represent H, OH, a C₁-C₁₂ alkyl, hydroxyalkyl or alkoxyl group;

R₂ and R₂′

independently represent H, OH, or an N-acetamido group, or C₁-C₁₂ alkyl alkyl group;

R₄ and R₄′

independently represent H or OH;

R₆ and R₆′

independently represent H, CH₂OH, CH3, or COOH;

R₇ represent H or OH;

R₀ represents H, methyl, or a straight chain or branched saturated or unsaturated carbon chain containing 9-12 carbon atoms, optionally substituted with a phenyl, hydroxyl or cyclohexyl group.