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1. (WO2007003594) SCREENING METHODS FOR INHIBITORS OF THE METALLOPROTEASE MEPRIN
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Claims

1. A method of treating a human patient having need of maintaining or increasing BNP levels by administering a therapeutically effective amount of a compound modulating or inhibiting meprin activity.

2. The method according to claim 1, wherein the therapeutically effective amount of a compound modulating or inhibiting meprin activity is administered in combination with a therapeutically effective amount of an inhibitor of dipeptidyl peptidase IV.

3. A method of treating a disease or disorder associated with decreased or insufficient levels of BNP in mammals and humans comprising administering to a subject in need thereof a therapeutically effective amount of a compound modulating or inhibiting meprin activity or downregulating meprin expression.

4. The method according to claim 3, wherein the therapeutically effective amount of a compound modulating or inhibiting meprin activity or downregulating meprin expression is administered in combination with a therapeutically effective amount of an inhibitor of dipeptidyl peptidase IV.

5. A method of treating or preventing cardiovascular and/or renal disorders or diseases in mammals and humans comprising administering to a subject in need thereof a therapeutically effective amount of a compound modulating or inhibiting meprin activity.

6. The method according to claim 5, wherein the therapeutically effective amount of a compound modulating or inhibiting meprin activity is administered in combination with a therapeutically effective amount of an inhibitor of dipeptidyl peptidase IV.

7. A method according to any of claims 1 to 6, wherein the meprin is Meprin A.

8. A method according to claim 5, wherein the cardiovascular disorder or disease is selected from the group consisting of atherosclerosis, peripheral vascular disease, cerebral vascular disease, cardiac ischemia; ischemic heart disease, acute coronary syndrome, stable and unstable angina, acute myocardial infarction, post myocardial infarction, acute and chronic heart failure, including congestive heart failure; peripheral occlusive disease; ischemic stroke; hypertension, including essential hypertension and secondary forms of hypertension such as renal hypertension and/or pulmonary hypertension.

9. A method according to claim 5, wherein the renal disorder or disease is selected from acute renal failure and chronic renal failure.

10. A method according to claim 5 or 8 further comprising administering to said subject a thera- peutically effective amount of a NEP inhibitor.

11. An in vitro screening method to identify therapeutic agents useful in the treatment of cardiovascular and/or renal disorders or diseases in a mammal, wherein
(i) the method screens for therapeutic agents, which specifically bind to a meprin metalloprote- ase, said method comprising the steps of
a) providing a test compound;
b) contacting the test compound with said meprin metalloprotease for a sufficient time and under suitable conditions for binding; and
c) detecting binding of the meprin metalloprotease to the test compound, thereby identifying a therapeutic agent which specifically binds the meprin metalloprotease; and/or
(ii) the method screens for therapeutic agents, which modulate the activity of a meprin metalloprotease, said method comprising the steps of
a) providing a test compound;
b) contacting the test compound with said meprin metalloprotease; and
c) assaying a biological activity of said meprin metalloprotease,
wherein a test compound which increases said biological activity is identified as a potential therapeutic agent for increasing the activity of the meprin metalloprotease, and wherein a test compound which decreases said biological activity is identified as a potential therapeutic agent for decreasing the meprin activity; and/or
(iii) the method screens for therapeutic agents, which inhibit or decrease the activity of a meprin metalloprotease, said method comprising the steps of :
a) providing a test compound;
b) contacting the test compound with the meprin metalloprotease;
c) assaying a biological activity of the meprin metalloprotease in presence and absence and/or at different concentrations of said test compound,
d) optionally assaying said biological activity in the presence of a compound known to be a regulator of the meprin metalloprotease,
wherein a test compound which decreases or inhibits said biological activity is identified as a potential therapeutic agent for inhibiting or decreasing meprin activity.

12. A method according to claim 11 , wherein the biological activity of the meprin metalloprotease is assayed by measuring the influence of the test compound on the activity of the meprin metalloprotease in the presence of a suitable substrate for the meprin metalloprotease.

13. A method according to claim 12, wherein the substrate is BNP.

14. A method according to claim 13, wherein the biological activity of the meprin metalloprotease is assayed by measuring the degree of BNP degradation.

15. A method according to claim 11 , wherein the compound known to be a regulator of the meprin metalloprotease is actinonin.

16. A method according to claim 11 , wherein the method screens for therapeutic agents which inhibit or decrease the activity of a meprin metalloprotease, comprising the steps of
a) providing a test compound;
b) contacting the test compound with said meprin metalloprotease in the presence of BNP, c) measuring the degree of BNP degradation, and
d) comparing the degree of BNP degradation by said meprin metalloprotease in presence and absence and/or at different concentrations of said test compound,
wherein a test compound which induces a lower degree of BNP degradation is identified as a potential therapeutic agent for inhibiting or decreasing meprin activity.

17. A method according to any of the claims 11 to 16, wherein the meprin metalloprotease is meprin A.

18. A method according to claim 17, wherein the meprin A is of human, mouse or rat origin.

19. A method according to claim 17, wherein the meprin A has at least 90% identity to an amino acid sequence as displayed in SEQ ID NO: 1 , SEQ ID NO: 2, or SEQ ID NO: 3.

20. A method according to claim 13, 14 or 16, wherein the BNP used is mouse BNP or human
BNP, preferably mouse BNP(I -32) or human BNP(I -32).

21. A method according to claim 20, wherein the BNP has the amino acid sequence as displayed in SEQ ID NO: 4 or SEQ ID NO: 6.

22. A method according to any of claims 11 to 21 , wherein the meprin metalloprotease comprises at least one meprin fusion protein.

23. A method according to any of claims 11 to 21 , wherein the step of contacting is in or at the surface of a cell.

24. A method according to any of claims 11 to 21 , wherein the step of contacting is in a cell- free system.

25. A method according to any of claims 11 to 21 , wherein the meprin metalloprotease is pro- vided i) as isolated protein, ii) in the form of a membrane preparation bearing said meprin protein, or iii) in the form of an intact cell or cell extracts comprising said meprin protein.

26. A method according to any of claims 11 to 21 , wherein the meprin metalloprotease is coupled to a detectable label.

27. A method according to any of claims 11 to 21 , wherein the test compound is coupled to a detectable label.

28. A method according claim 13, 14, 16, 20 or 21 , wherein the BNP is coupled to a detectable label.

29. A method according to any of claims 11 to 21 , wherein the meprin metalloprotease is attached to a solid support.

30. A method according to any of claims 11 to 29, wherein a test compound being identified as a potential therapeutic agent for inhibiting or decreasing meprin activity is further tested for its ability to inhibit or decrease the activity of a dipeptidyl peptidase IV, comprising the steps of a) contacting the test compound with said dipeptidyl peptidase IV,
b) assaying a biological activity of said dipeptidyl peptidase IV in the presence and absence and/or at different concentrations of said test compound,
wherein a test compound which decreases or inhibits said biological activity is identified as a potential therapeutic agent for inhibiting or decreasing meprin and dipeptidyl peptidase IV activity.

31. A compound for use in therapy that modulates, inhibits or decreases the activity of a meprin metalloprotease, wherein said compound is identified by any of the methods of claims 11 to 30.

32. A compound according to claim 31 for the treatment and/or prophylaxis of cardiovascular or renal disorders or diseases.

33. A method of treating or preventing cardiovascular disorders or diseases in mammals and humans comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to claim 31.

34. Use of a compound inhibiting, modulating or decreasing meprin activity for the manufacture of a medicament for the prophylaxis and/or treatment of cardiovascular disorders or diseases and/or renal diseases or disorders.

35. Use of a compound according to claim 34, wherein the compound further inhibits or decreases dipeptidyl peptidase IV activity.

36. Use according to Claim 34, wherein the cardiovascular disorder or disease is selected from the group consisting of atherosclerosis, peripheral vascular disease, cerebral vascular disease, cardiac ischemia; ischemic heart disease, acute coronary syndrome, stable and unstable angina, acute myocardial infarction, post myocardial infarction, acute and chronic heart failure, including congestive heart failure; peripheral occlusive disease; ischemic stroke; hypertension, including essential hypertension and secondary forms of hypertension such as renal hypertension and/or pulmonary hypertension.

37. Use according to claim 34, wherein the renal disorder or disease is selected from acute renal failure and chronic renal failure.

38. Use according to any of the claims 34 to 37, wherein the compound is identified by the method according to any of the claims 11 to 30.

39. Use according to claim 38, wherein said compound is
a) a small molecule,
b) an RNA molecule,
c) an antisense oligonucleotide,
d) a polypeptide,
e) an antibody,
f) a small interfering RNA molecule (siRNA), or
g) a ribozyme.

40. Use according to any of the claims 34 to 39, wherein the medicament further comprises one or more additional pharmaceutical agents.

41. Use according to claim 40, wherein said additional pharmaceutical agent is an cardiovascular-active agent selected from the group consisting of beta-blockers, calcium channel blockers, diuretics, renin inhibitors, ACE inhibitors, AT-1 receptor antagonists, ET receptor antagonists, NEP inhibitors, SEP inhibitors, ECE inhibitors, dipeptidyl peptidase IV inhibitors and nitrates.

42. A pharmaceutical composition comprising a therapeutically effective amount of a compound which inhibits or decreases meprin activity in combination with a pharmaceutically acceptable carrier.

43. A pharmaceutical composition according to claim 42, wherein the compound is identified by the method according to any of the claims 11 to 30.

44. A pharmaceutical composition according to one of the claims 42 or 43 further comprising one or more additional pharmaceutical agents.

45. The pharmaceutical composition of claim 44, wherein said additional pharmaceutical agent is an cardiovascular-active agent selected from the group consisting of beta-blockers, calcium channel blockers, diuretics, renin inhibitors, ACE inhibitors, AT-1 receptor antagonists, ET receptor antagonists, NEP inhibitors, SEP inhibitors, ECE inhibitors, dipeptidyl peptidase IV inhibitors and nitrates.