Synthetic peptide inhibitors of thrombin and thrombin activation of protease activated receptors 1 and 4
View Patent ↗The invention relates to synthetic peptide analogs of D-Arg-Oic-Pro-Gly-Phe and methods of their use to inhibit human platelet aggregation, thrombosis and cell activation mediated by PAR1 and PAR4.
1. A compound comprising an amino acid sequence of formula
A 1 -A 2 -A 3 -A 4 -A 5
Wherein
A 1 is selected from the group consisting of D-arginine (r) and L-arginine (R);
A 2 is selected from the group consisting of (2S, 3aS, 7aS)-octahydroindole-2-carboxlic acid (Oic) and P;
A 3 is selected from the group consisting of Oic and P;
A 4 is selected from the group consisting of D-alanine (a) and D-serine (s); and
A 5 is selected from the group consisting of L-[para-methyl]-phenylalanine(F(p-me)), L-[para-bromo]-phenylalanine(F(p-Br)), L-[para-iodo]-phenylalanine(F(p-I)), and L-[para-nitro]-phenylalanine F(p-NO 2 )).
2. The compound of claim 1 that comprises at least one blocking group.
3. The compound of claim 2 that is amidated at a C-terminus.
4. The compound of claim 1 that consists of A 1 -A 2 -A 3 -A 4 -A 5 .
5. The compound of claim 4 selected from the group consisting of rOicPGF, rOicPaF(p-Me), rOicPsF(p-Me), rOicPaF(p-Br), rOicPaF(p-I) and rOicPaF(p-NO 2 ).
6. The compound of claim 1 comprising a formula selected from the group consisting of rOicPGF, rOicPaF(p-Me), r OicPsF(p-Me), rOicPaF(p-Br), rOicPaF(p-I) and rOicPaF(p-NO 2 ).
7. The compound of claim 6 comprising the formula rOicPaF(p-Me) or rOicPsF(p-Me).
8. A method of inhibiting thrombin-induced platelet aggregation comprising administering an effective amount of a compound of claim 1 to platelets.
9. A method of inhibiting a thrombin-mediated activity comprising administering an effective amount of a compound of claim 1 to a cell.
10. The method of claim 9 wherein the activity is coagulation.
11. The method of claim 10 wherein the activity is platelet aggregation.
12. The method of claim 9 wherein the activity is cell motility.
13. The method of claim 12 wherein said cell is a cancer cell.
14. The method of claim 9 wherein the activity is cell adhesion.
15. The method of claim 9 wherein the activity is calcium mobilization.
16. A method of inhibiting thrombin activation of mammalian cells having thrombin receptors, said method comprising contacting said cells with an effective amount of a compound of claim 1 .
17. The method of claim 16 wherein said compound comprises a sequence selected from the group consisting of rOicPGF, rOicPaF(p-Me), r OicPsF(p-Me), rOicPaF(p-Br), rOicPaF(p-I) and rOicPaF(p-NO 2 ).
18. A method of inhibiting thrombin-induced platelet activation, platelet aggregation or thrombosis in a mammal comprising administering an effective amount of a compound of claim 1 to said mammal.
19. The method of claim 18 wherein said mammal is a human.
20. A pharmaceutical composition comprising a compound of claim 1 , and optionally at least one pharmaceutically acceptable carrier or excipient.
21. The composition of claim 20 comprising rOicPaF(p-Me).
22. A method of treating acute coronary syndrome in an individual in need thereof comprising administering an effective amount of a compound of claim 1 .
23. The method of claim 22 wherein said compound is administered orally.
24. The compound of claim 1 wherein A 1 -A 2 -A 3 -A 4 -A 5 is located at the N-terminus of the peptide.
25. A method of inhibiting a thrombin-mediated activity comprising administering an effective amount of a compound of claim 24 to a cell.
26. The method of claim 25 wherein said compound is administered orally.
27. A compound of formula MAP4-A 1 -A 2 -A 3 -A 4 -A 5 ,
wherein MAP4 is a four-branched peptide consisting of a β-alanine core with a single lysine attached at its amino terminal end followed by two additional lysines;
A 1 is selected from the group consisting of D-arginine (r) and L-arginine (R);
A 2 is selected from the group consisting of (2S, 3aS, 7aS)-octahydroindole-2-carboxlic acid (Oic) and P;
A 3 is selected from the group consisting of Oic and P;
A 4 is selected from the group consisting of D-alanine (a) and D-serine (s); and
A 5 is selected from the group consisting of L-[para-methyl]-phenylalanine(F(p-me)), L-[para-bromo]-phenylalanine(F(p-Br)), L-[para-iodo]-phenylalanine(F(p-I)), and L-[para-nitro]-phenylalanine F(p-NO 2 )); and
wherein at least one non-naturally occurring amino acid residue is present in A 1 -A 2 -A 3 -A 4 -A 5 .
28. A method of inhibiting a thrombin-mediated activity comprising administering an effective amount of a compound of claim 26 to a cell.
29. The method of claim 28 wherein said compound is administered orally.
30. The compound of claim 1 that is an inhibitor of at least one thrombin-mediated activity.
31. The compound of claim 30 wherein the activity is selected from the group consisting of thrombin-induced platelet aggregation, thrombin-induced calcium mobilization, thrombin-mediated coagulation, thrombin-induced cell motility and thrombin-induced cell adhesion.
32. The compound of claim 30 wherein the activity is thrombin-induced platelet aggregation.
33. The compound of claim 30 wherein the activity is thrombin-induced calcium mobilization.
34. The compound of claim 30 wherein the activity is thrombin-mediated coagulation.
35. The compound of claim 32 that is an inhibitor of thrombin-induced calcium mobilization.
36. The compound of claim 32 that is an inhibitor of thrombin-mediated coagulation.
37. The compound of claim 32 wherein the activity is thrombin-induced cell motility.
38. The compound of claim 32 wherein the activity is thrombin-induced cell adhesion.