Superoxide dismutase mimic

• US 5,227,405 A
• Filed: 09/28/1988
• Issued: 07/13/1993
• Est. Priority Date: 03/31/1987
• Status: Expired due to Term

• Current Assignee: Duke University
• Original Assignee: Duke University
• Inventors: Beyer, Wayne F., Fridovich, Irwin, Darr, Douglas J.
• Primary Examiner(s): Patterson, Jr., Charles L.
• Application Number:
  US07/250,367
• Time in Patent Office: 1,749 Days
• Field of Search: 435/189, 424/94.4, 424/DIG. 6, 514/616, 514/516, 514/612, 564/153, 564/114, 564/152
• US Class Current: 514/612
• CPC Class Codes: A01N 1/0226 Physiologically active agents, i.e. substances affecting physiological processes of cells and tissue to be preserved, e.g. anti-oxidants or nutrients
• CPC Class Codes: A01N 25/32 Ingredients for reducing the noxious effect of the active substances to organisms other than pests, e.g. toxicity reducing compositions, self-destructing compositions
• CPC Class Codes: A01N 37/30 containing the groups —CO—N< and , both being directly attached by their carbon atoms to the same carbon skeleton, e.g. H2N—NH—CO—C6H4—COOCH3; Thio-analogues thereof
• CPC Class Codes: A01N 43/40 six-membered rings
• CPC Class Codes: A01N 59/16 Heavy metals; Compounds thereof
• CPC Class Codes: C07F 13/005 Compounds without a metal-carbon linkage
• CPC Class Codes: Y10S 424/06 Chelate

Abstract

The invention relates to a water-soluble complex formed between a chelating agent and manganese and pharmaceutical compositions thereof. The complex is a low molecular weight mimic of superoxide dismutase. The invention further relates to a method of using the complex comprising treating plant and animal cells with an amount of the complex sufficient to reduce or prevent superoxide radical-induced toxicity.

Claims

1. A low molecular weight mimic of superoxide dismutase activity comprising a water solube complex formed between a chelating agent and manganese, wherein said chelating agent is a hydroxamate-type siderophore, or analog or derivative thereof capable of forming said complex.

2. A mimic of superoxide dismutase activity according to claim 1 wherein said manganese has an actual valence of three.

3. A mimic of superoxide dismutase activity according to claim 1 wherein said chelating agent is desferrioxamine or analog or derivative thereof capable of forming said complex.

4. A mimic of superoxide dismutase activity according to claim 1 in essentially pure form.

5. A pharmaceutical composition comprising, as an active ingredient, a water-soluble complex formed between a chelating agent and manganese, wherein said chelating agent is a hydroxamate-type siderophore, or analog or derivative thereof capable of forming said complex, which complex is present in an amount sufficient to reduce or prevent superoxide radical-induced toxicity, together with a pharmaceutically acceptable carrier.

6. A pharmaceutical composition according to claim 5 wherein said composition is in the form of a cream or a lotion for topical application.

7. A pharmaceutical composition according to claim 5 wherein said composition is in the form of a tablet, capsule or sterile injectible solution.

8. A pharmaceutical composition according to claim 5 wherein said chelating agent is desferrioxzamine or analog or derivative thereof capable of forming said complex.

9. A pharmaceutical compositon according to claim 5 wherein said manganese has an actual valence of three.

10. A low molecular mimic of superoxide dismutase produced by a method comprising the steps of:

a) combining a manganese (IV) salt with a chelating agent in water or an aqueous buffer, wherein said chelating agent is a hydroxamate-type siderophore, or analog or derivative thereof capable of complexing with manganese;

b) stirring said combination until reaction between said chelating agent and manganese is complete; and

c) removing residual manganese salt from a resulting solution.

11. A mimic of superoxide dismutase activity produced by the method according to claim 10 further comprising the step of heating said resulting solution at approximately 100°

C. for approximately 15 minutes.

18. A mimic of superoxide dismutase activity produced by the method according to claim 10, wherein said aqueous buffer has a pH of about 6 to 8.

12. A low molecular weight mimic of superoxide dismutase activity produced by a method comprising the steps of:

a) combinting a manganese(II) salt with a chelating agent in a neutral oxygenated aqueous solution, wherein said chelating agent is a hydroxamate-type siderophore, or analog or derivative thereof capable of complexing with manganese;

b) stirring said combination until reaction between said chelating agent and manganese is complete; and

c) removing any residual manganese salt from a resulting solution.

13. A mimic of superoxide dismutase activity produced by a method comprising the steps of:

a) combing a manganese salt, in which salt manganese has a valence of III, V, VI or VII, with a chelating agent under such conditions that manganese achieves an actual valence of three, wherein said chelating agent is a hydroxamate-type siderophore, or analog or derivative thereof capable of complexing with manganese;

b) stirring said combination until reaction between said chelating agent and said manganese is complete; and

c) removing any residual manganese salt from a resulting solution.

36. The mimic of superoxide dismutase activity according to claim 13 wherein said manganese of said manganese salt has a valence of III.

14. A low molecular weight mimic of superoxide dismutase activity produced by a method comprising the steps of:

a) combining a hydroxamate-type siderophore chelating agent and a reducing agent in an aqueous solution and adjusting the pH to about 8.5;

b) adding to said solution of step (a) a manganese (IV) salt;

c) allowing said chelating agent to chelate the manganese of said manganese (IV) salt; and

d) removing residual manganese salt from a solution resulting from step (c) above.

15. The low molecular weight mimic of superoxide dismutase activity according to claim 14 wherein said reducing agent is ascorbic acid, or a salt thereof.

16. The low molecular weight mimic of superoxide dismutase activity according to claim 14 wherein said chelating agent is desferrioxamine.

17. The low molecular weight mimic of superoxide dismutase activity according to claim 14 wherein said manganese (IV) salt is MnO₂.

31. A pharmaceutical composition comprising, as an active ingredient, said low molecular weight mimic according to claim 14 which mimic is present in an amount sufficient to reduce or prevent superoxide radical induced toxicity, together with a pharmaceutically acceptable carrier.

32. A pharmaceutical composition according to claim 31, wherein said reducing agent is ascorbic acid, or a salt thereof.

33. A pharmaceutical composition according to claim 31, wherein said chelating agent is desferrioxamine.

34. The pharmaceutical composition according to claim 31 wherein said composition is in the form of a cream or a lotion suitable for topical application.

35. The pharmaceutical composition according to claim 31 wherein said composition is in the form of a tablet, capsule or sterile injectable solution.

19. A method of protecting cells from the toxicity of superoxide radicals comprising treating said cells with an amount of a water-soluble complex formed between a hydroxamate-type siderophore chelating agent, or analog or derivative thereof capable of complexing with manganese, and manganese sufficient to reduce or prevent superoxide radical-induced toxicity, which complex is produced by a method comprising the steps of:

a) combining said hydroxamate-type siderophore chelating agent, or analog or derivative thereof, and a reducing agent in an aqueous solution and adjusting the pH to about 8.5;

b) adding to said solution of step (a) a manganese (IV) salt;

c) allowing said chelating agent to chelate the manganese of said manganese (IV) salt; and

d) removing any residual manganese salt from soltuion resulting from step (c) above.

20. The method according to claim 19 wherein said cells are mammalian cells.

21. The method according to claim 19 wherein said cells are plant cells.

22. The method according to claim 19 wherein said superoxide-radicals are the result of a reperfusion injury.

23. The method according to claim 19 wherein said superoxide-radicals are the result of an inflammatory disease.

24. The method according to claim 19 wherein said reducing agent is ascorbic acid, or a salt thereof.

25. The method according to claim 19 wherein said manganese (IV) salt is MnO₂.

26. The method according to claim 19 wherein said hydroxamate type siderophore chelating agent is desferrioxamine or analog or derivative thereof capable of forming said complex.

27. A method of inhibiting damage due to auto-oxidation of a substance with the subsequent formation of O₂ comprising adding to said substance an amount of a water-soluble complex formed between a hydroxamate-type siderophore chelating agent, or analog or derivative thereof capable of complexing with manganese, and manganese sufficient to reduce or prevent oxidation damage, which complex is produced by a method comprising the steps of:

a) combining said hydroxamate-type siderophore chelating agent, or analog or derivative thereof, and a reducing agent in an aqueous solution and adjusting the pH to about 8.5;

b) adding to said solution of step (a) a manganese (IV) salt;

c) allowing said chelating agent to chelate the manganese of said manganese (IV) salt; and

d) removing any residual mangese salt from a solution resulting from step (c) above.

28. The method according to claim 27 wherein said reducing agent is ascorbic acid, or a salt thereof.

29. The method according to claim 27 wherein said manganese (IV) salt is MnO₂.

30. The method according to claim 27 wherein said hydroxamate-type siderophore is desferrioxamine, or analog or derivative thereof capable of forming said complex.

37. A mimic of superoxide dismutase activity comprising an atom of manganese having an actual valence of 3 complexed with three hydroxamate groups.

39. A pharmaceutical composition comprising as an active ingredient said mimic according to claim 37, which mimic is present in an amount sufficient to reduce or prevent superoxide radical induced toxicity, together with a pharmaceutically acceptable carrier.

40. A pharmaceutical composition according to claim 39 wherein said hydroxamate groups are derived from a single molecule of desferrioxamine.

41. A pharmaceutical composition according to claim 39 wherein said composition is in the form of a cream or a lotion suitable for topical application.

42. A pharmaceutical composition according to claim 39 wherein said composition is in the form of a tablet, capsule or sterile injectable solution.

38. The mimic of superoxide dismutase activity according to claiim 37 wherein said hydroxamate groups are derived from a single molecule of desferrioxamine.

43. A complex of a desferrioxamine and manganese.

44. The complex according to claim 43, wherein the desferrioxamine is a trihydroxamic acid.

45. The complex according to claim 43, wherein the desferrioxamine is of the formula, ##STR1##

46. A stable aqueous solution of a complex of a desferrioxamine and manganese, containing alkali in amount sufficient to bring the pH to about 6.

47. The solution according to claim 46, wherein the desferrioxamine is a trihydroxamic acid.

48. The solution according to claim 47, wherein the desferrioxamine is of the formula ##STR2##