Therapeutic procedures

• US 20060217359A1
• Filed: 03/06/2006
• Published: 09/28/2006
• Est. Priority Date: 09/05/2003
• Status: Abandoned Application

• Current Assignee: The Scripps Research Institute
• Original Assignee: The Scripps Research Institute
• Inventors: Lerner, Richard A., Wentworth, Paul
• Application Number:
  US11/369,206
• Publication Number:
  US 20060217359A1
• Time in Patent Office: Days
• US Class Current: 514/178
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/10 Antimycotics
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• 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: C07C 49/523 containing —CHO groups
• CPC Class Codes: C07C 49/757 containing —CHO groups
• CPC Class Codes: C07C 59/353 containing rings
• CPC Class Codes: C07D 313/04 not condensed with other rings
• CPC Class Codes: C07J 61/00 Steroids in which the cyclopenta(a)hydrophenanthrene skeleton has been modified by contraction of only one ring by one or two atoms
• CPC Class Codes: C07J 9/00 Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of more than two carbon atoms, e.g. cholane, cholestane, coprostane
• CPC Class Codes: C07J 9/005 containing a carboxylic function directly attached or attached by a chain containing only carbon atoms to the cyclopenta[a]hydrophenanthrene skeleton

Abstract

As illustrated herein, cholesterol is oxidized when it is present in atherosclerotic plaques. This reaction generates cytotoxic cholesterol oxidation or ozonation products. The present application is directed to the products of cholesterol ozonation, binding entities directed against such products, and methods of using such binding entities and cytotoxins to treat a variety of diseases.

Claims

1. An isolated ozonation product of cholesterol that can be cytotoxic to a prokaryotic or eukaryotic cell.

2. The ozonation product of claim 1 that can cause macrophage lipid uptake or foam cell formation from macrophages.

3. The ozonation product of claim 1 that can change the secondary structure of a protein in a low density lipoprotein.

4. The ozonation product of claim 3, wherein the protein is apoprotein B₁₀₀.

5. The ozonation product of claim 1 having formula 4a:

6. The ozonation product of claim 1 having formula 5a:

7. The ozonation product of claim 1 having any one of formulae 6a-15a, 7c or a combination thereof.

8. A composition comprising a carrier and an isolated ozonation product of cholesterol that can be cytotoxic to a prokaryotic or eukaryotic cell.

9. The composition of claim 8, wherein the ozonation product comprises formula 4a:

10. The composition of claim 8, wherein the ozonation product comprises formula 5a:

11. The composition of claim 8, wherein the ozonation product comprises a compound having any one of formulae 6a-15a, 7c or a combination thereof

12. An isolated binding entity that can bind to an ozonation product of cholesterol.

13. The binding entity of claim 12, wherein the ozonation product of cholesterol is a compound of formula 4a:

14. The binding entity of claim 12, wherein the ozonation product of cholesterol is a compound of formula 5a:

15. The binding entity of claim 12, wherein the antibody can also bind to a compound having any of formulae 6a, 7a, 7c, 8a, 9a, 10a, 11a, 12a, 13a, 14a or 15a:

16. The binding entity of claim 12, wherein the binding entity was raised against a hapten having formula 13a, 14a or 15a:

17. The binding entity of claim 12, wherein the binding entity is an antibody.

18. The binding entity of claim 17, wherein the isolated antibody is a derived from hybridoma KA1-11C5 or KA1-7A6 having ATCC Accession No. PTA-5427 or PTA-5428.

19. The binding entity of claim 17, wherein the isolated antibody is a derived from hybridoma KA2-8F6 or KA2-1E9, having ATCC Accession No. PTA-5429 and PTA-5430.

20. The binding entity of claim 12, wherein the binding entity is linked a therapeutic agent.

21. The binding entity of claim 20, wherein the therapeutic agent can reduce an atherosclerotic lesion or prevent further occlusion of the artery.

22. The binding entity of claim 20, wherein the therapeutic agent is an anti-oxidant, anti-inflammatory agent, drug, small molecule, peptide, polypeptide or nucleic acid.

23. A binding entity linked to an ozonation product of cholesterol, wherein the ozonation product of cholesterol is cytotoxic to a prokaryotic or eukaryotic cell.

24. The binding entity of claim 23, wherein the binding entity is an antibody.

25. The binding entity of claim 23, wherein the ozonation product of cholesterol is a compound of any one of formulae 4a-15a or 7c:

26. A marker for treating or preventing atherosclerotic lesions comprising an ozonation product of cholesterol having formula 4a or formula 5a:

27. A method for treating atherosclerosis in a patient comprising administering to the patient a binding agent that can bind to an ozonation product of cholesterol.

28. The method of claim 27, wherein the ozonation product of cholesterol is a compound having any one of formulae 4a-15a or 7c:

29. The method of claim 27, wherein binding agent does not generate a reactive oxygen species.

30. The method of claim 27, wherein the binding entity is linked a therapeutic agent.

31. The method of claim 30, wherein the therapeutic agent can help slow the growth or reduce the size of an atherosclerotic lesion.

32. The method of claim 30, wherein the therapeutic agent is an anti-oxidant, anti-inflammatory agent, drug, small molecule, peptide, polypeptide or nucleic acid.

33. A method for killing a target cell in a patient comprising administering to the patient a binding agent that can bind to the target cell, wherein the binding agent is linked to an ozonation product of cholesterol.

34. The method of claim 33, wherein the binding entity is an antibody.

35. The method of claim 33, wherein the antibody can generate a reactive oxygen species.

36. The method of claim 33, wherein the antibody is also linked to a compound that can generate singlet oxygen.

37. The method of claim 36, wherein the compound that can generate singlet oxygen is an endoperoxide.

38. The method of claim 36, wherein the compound that can generate singlet oxygen is an anthracene-9,10-dipropionic acid endoperoxide.

39. The method of claim 36, wherein the compound that can generate singlet oxygen is a pterin, flavin, hematoporphyrin, tetrakis(4-sulfonatophenyl) porphyrin, bipyridyl ruthenium(II) complex, rose Bengal dye, quinone, rhodamine dye, phthalocyanine, hypocrellin, rubrocyanin, pinacyanol or allocyanine.

40. A method for removing cytotoxic cholesterol ozonation products from a mammal comprising separating the cytotoxic cholesterol ozonation products from bodily fluids of the mammal using a binding entity or an antibody that can bind to an ozonation product of cholesterol.

41. The method of claim 40, wherein the ozonation product of cholesterol is a compound of formula 4a:

42. The method of claim 40, wherein the ozonation product of cholesterol is a compound of formula 5a:

43. The method of claim 40, wherein the ozonation product of cholesterol is a compound having any one of formulae 6a-15a or 7c:

44. The method of claim 40, wherein the ozonation product is removed from circulating blood of the mammal.

45. The method of claim 40, wherein the ozonation product is removed ex vivo from blood of the mammal.

46. The method of claim 40, wherein the binding entity or the antibody is administered in a localized manner to the localized tissues.

47. A method of treating or preventing cancer in a mammal comprising administering to the mammal an antibody linked to a cytotoxic ozonation product of cholesterol, wherein the antibody can bind to a cancer cell.

51. The method of any one of claims 47-49, wherein the antibody can bind a cholesterol ozonation product having any one of formulae 4a-15a or 7c:

48. A method of treating or preventing an inappropriate immune response in a mammal comprising administering to the mammal an antibody linked to a cytotoxic ozonation product of cholesterol, wherein the antibody can bind to an immune cell involved in the inappropriate immune response.

49. A method for identifying an agent that modulates the production of a reactive oxygen species from an antibody, comprising:

(a) combining an antibody and a candidate agent;

(b) determining the amount of reactive oxygen species formed; and

(c) comparing the amount of reactive oxygen species formed with a standard value obtained by determining the amount of reactive oxygen species formed from the antibody without the candidate agent.

50. The method of claim 49, wherein the reactive oxygen species is ozone.