G protein coupled receptor antagonists and methods of activating and inhibiting G protein coupled receptors using the same
The invention relates generally to G protein coupled receptors and in particular to agonists and antagonists of G protein receptors and methods of using the same.
1. A method of treating a cardiovascular disorder in a mammal, comprising administering to a mammal in need thereof in an amount sufficient to reduce the severity of the disorder a polypeptide comprising:
a) a first domain comprising a third intracellular loop, or a fragment thereof of a Protease-Activated Receptor (PAR); and
b) a second domain, attached to the first domain, wherein the second domain comprises a cell-penetrating, membrane-tethering moiety comprising: a lipid, a cholesterol, a phospholipid, a steroid, a sphingosine, a ceramide, an octyl-glycine, a 2-cyclohexylalanine, or a benzolylphenylalanine;
wherein the polypeptide is a PAR antagonist,
wherein the first domain does not comprise an extracellular portion of the PAR, and
wherein the first domain comprises at least 3 contiguous amino acid residues of the third intracellular loop.
2. The method of claim 1 , wherein the cardiovascular disorder is selected from the group consisting of arterial thrombosis, atherosclerosis and restenosis.
3. The method of claim 1 or 2 , wherein the PAR is Protease-Activated Receptor 1 (PAR-1) or PAR-4.
4. The method of claim 3 , wherein the PAR antagonist comprises SEQ ID NO: 3 or SEQ ID NO: 4.
5. The method of claim 3 , wherein the PAR antagonist comprises SEQ ID NO: 29.
6. The method of claim 1 , wherein the second domain comprises a palmitoyl group, a myristoyl group, or a pentadecanoyl group.
7. The method of claim 1 , wherein the second domain comprises a lipid.
8. The method of claim 1 , wherein the polypeptide is infused into a vascular lumen.
9. The method of claim 8 , wherein the vascular lumen is the lumen of a jugular or peripheral vein.
10. The method of claim 1 , wherein the polypeptide is infused into a perivascular space.
11. The method of claim 1 , wherein the polypeptide is administered transdermally, subdermally, or subcutaneously.
12. The method of claim 1 , wherein the polypeptide is administered into the lung tissue of said mammal.
13. The method of claim 1 , wherein said mammal is human.
14. The method of claim 1 , wherein said PAR antagonist comprises an amino acid sequence selected from the group consisting of: SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 9, and SEQ ID NO: 10.
15. The method of claim 1 , wherein the second domain comprises a cholesterol.
16. The method of claim 1 , wherein PAR antagonist is P1pal-7.
17. A method of treating atherosclerosis in a mammal, comprising administering to a mammal in need thereof in an amount sufficient to reduce the severity of atherosclerosis a polypeptide comprising:
a) a first domain comprising a third intracellular loop, or a fragment thereof of a Protease-Activated Receptor (PAR); and
b) a second domain, attached to the first domain, wherein the second domain comprises a cell-penetrating, membrane-tethering moiety comprising: a lipid, a cholesterol, a phospholipid, a steroid, a sphingosine, a ceramide, an octyl-glycine, a 2-cyclohexylalanine, or a benzolylphenylalanine;
wherein said polypeptide is a PAR antagonist,
wherein the first domain does not comprise an extracellular portion of the PAR, and
wherein the first domain comprises at least 3 contiguous amino acid residues of the third intracellular loop.
18. The method of claim 17 , wherein the PAR is PAR-1 or PAR-4.
19. The method of claim 18 , wherein the PAR antagonist comprises SEQ ID NO: 3 or SEQ ID NO: 4.
20. The method of claim 18 , wherein the PAR antagonist comprises SEQ ID NO: 29.
21. The method of claim 17 , wherein the second domain comprises a palmitoyl group, a myristoyl group, or a pentadecanoyl group.
22. The method of claim 21 , wherein the polypeptide is P1pal-7.
23. The method of claim 17 , wherein the second domain comprises a lipid.
24. The method of claim 17 , wherein said polypeptide is infused into a vascular lumen.
25. The method of claim 24 , wherein the vascular lumen is the lumen of a jugular or peripheral vein.
26. The method of claim 17 , wherein the polypeptide is infused into a perivascular space.
27. The method of claim 17 , wherein the polypeptide is administered transdermally, subdermally, or subcutaneously.
28. The method of claim 17 , wherein the polypeptide is administered into the lung tissue of said mammal.
29. The method of claim 17 , wherein said mammal is human.
30. The method of claim 17 , wherein the second domain comprises a cholesterol.