G protein coupled receptor agonists and antagonists and methods of use
The invention relates generally to G protein coupled receptors (GPCRs) and in particular to GPCR agonists and antagonists, use of these compounds and their pharmaceutical compositions, e.g., in the treatment, modulation and/or prevention of physiological conditions associated with GPCRs, such as in treating conditions in which chemokine receptors play a role, e.g., sepsis, arthritis, inflammation and autoimmune diseases.
1. A chimeric polypeptide comprising:
a) a first domain that is an amino acid sequence selected from the group consisting of YQKKLRSMTD (SEQ ID NO:24) and MGYQKKLRSMTD (SEQ ID NO:25) and
b) a second domain, attached to the first domain, wherein the second domain comprises a cell-penetrating, membrane-tethering hydrophobic moiety comprising a lipid, a cholesterol, a phospholipid, a steroid, a sphingosine, a ceramide, an octylglycine, a 2- cyclohexylalanine, or a benzolylphenylalanine,
wherein said chimeric polypeptide is an antagonist of a chemokine receptor.
2. The chimeric polypeptide of claim 1 , wherein the chemokine receptor is a CXC chemokine receptor.
3. The chimeric polypeptide of claim 1 , wherein the second domain is attached at the N-terminal end of the first domain.
4. The chimeric polypeptide of claim 1 , wherein the second domain comprises a lipid.
5. The chimeric polypeptide of claim 4 , wherein the second domain comprises a hydrophobic moiety that is selected from the group consisting of a: nonanoyl (C 9 ); capryl (C 10 ); undecanoyl (C 11 ); lauroyl (C 12 ); tridecanoyl (C 13 ); myristoyl (C 14 ); pentadecanoyl (C 15 ); palmitoyl (C 16 ); phytanoyl (methyl substituted C 16 ); heptadecanoyl (C 17 ); stearoyl (C 18 ); nonadecanoyl (C 19 ); arachidoyl (C 20 ); heniecosanoyl (C 21 ); behenoyl (C 22 ); trucisanoyl (C 23 ); and lignoceroyl (C 24 ) moiety.
6. The chimeric polypeptide of claim 4 , wherein the second domain comprises a palmitoyl moiety.
7. The chimeric polypeptide of claim 4 , wherein the second domain comprises a myristoyl (C 14 ) or pentadecanoyl (C 15 ) moiety.
8. The chimeric polypeptide of claim 1 , wherein the second domain comprises a lithocholic acid or a salt thereof.
9. The chimeric polypeptide of claim 1 , wherein the second domain comprises a hydrophobic moiety that is selected from the group consisting of a cholesterol, a phospholipid, a steroid, a sphingosine, a ceramide, an octyl-glycine, a 2- cyclohexylalanine, a benzolylphenylalanine, and a C 3 -C 8 fatty acid.
10. The chimeric polypeptide of claim 1 , wherein the second domain comprises a steroid.
11. A pharmaceutical composition comprising the chimeric polypeptide of claim 1 and a pharmaceutically acceptable carrier.
12. The chimeric polypeptide of claim 1 , wherein the second domain comprises a hydrophobic moiety that is selected from the group consisting of a phospholipid, a steroid, a sphingosine, a ceramide, an octylglycine, a 2-cyclohexylalanine, and a benzolylphenylalanine.
13. The chimeric polypeptide of claim 1 , wherein the second domain is attached to the first domain with an amide bond, a sulfhydryl, an amine, an alcohol, a phenolic group, or a carbon-carbon bond.
14. The chimeric polypeptide of claim 1 , wherein the second domain is attached at the C-terminal end of the first domain.
15. A chimeric polypeptide comprising:
(a) a first domain that comprises an amino acid sequence that is selected from the group consisting of ILKMKVKKPAV (SEQ ID NO: 28), VLATQAPRLPST (SEQ ID NO: 29), ATQAPRLPST (SEQ ID NO: 30, VLATGAPRLPST (SEQ ID NO:31), ATGAPRLPST(SEQ ID NO:32), and FLFRTKKKHPAV (SEQ ID NO: 17) and
(b) a second domain, attached to the first domain, wherein the second domain comprises a cell-penetrating, membrane-tethering hydrophobic moiety comprising a lipid, a cholesterol, a phospholipid, a steroid, a sphingosine, a ceramide, an octylglycine, a 2- cyclohexylalanine, or a benzolylphenylalanine;
wherein said chimeric polypeptide is an antagonist of a protease-activated receptor (PAR).
16. The chimeric polypeptide of claim 15 , wherein said PAR is PARI, PAR 2 , or PAR 4 .
17. The chimeric polypeptide of claim 15 , wherein the second domain comprises a lipid.
18. The chimeric polypeptide of claim 15 , wherein the second domain comprises a hydrophobic moiety that is selected from the group consisting of a: nonanoyl (C 9 ); capryl (C 10 ); undecanoyl (C 11 ); lauroyl (C 12 ); tridecanoyl (C 13 ); myristoyl (C 14 ); pentadecanoyl (C 15 ); palmitoyl (C 16 ); phytanoyl (methyl substituted C 16 ); heptadecanoyl (C 17 ); stearoyl (C 18 ); nonadecanoyl (C 19 ); arachidoyl (C 20 ); heniecosanoyl (C 21 ); behenoyl (C 22 ); trucisanoyl (C 23 ); and lignoceroyl (C 24 ) moiety.
19. The chimeric polypeptide of claim 15 , wherein the second domain is attached to the first domain with an amide bond, a sulfhydryl, an amine, an alcohol, a phenolic group, or a carbon-carbon bond.
20. The chimeric polypeptide of claim 15 , wherein the second domain comprises a palmitoyl moiety.
21. The chimeric polypeptide of claim 15 , wherein the second domain comprises a lithocholic acid or a salt thereof.
22. The chimeric polypeptide of claim 15 , wherein the second domain comprises a hydrophobic moiety that is selected from the group consisting of a cholesterol, a phospholipid, a steroid, a sphingosine, a ceramide, an octyl-glycine, a 2- cyclohexylalanine, a benzolylphenylalanine, and a C 3 -C 8 fatty acid.
23. The chimeric polypeptide of claim 15 , wherein the second domain comprises a steroid.
24. The chimeric polypeptide of claim 15 , wherein the second domain comprises a hydrophobic moiety that is selected from the group consisting of a phospholipid, a steroid, a sphingosine, a ceramide, an octylglycine, a 2-cyclohexylalanine, and a benzolylphenylalanine.
25. A pharmaceutical composition comprising the chimeric peptide of claim 15 and a pharmaceutically acceptable carrier.
26. A chimeric polypeptide comprising:
(a) a first domain that comprises an amino acid sequence that is selected from the group consisting of ILYSRVGRSVTD (SEQ ID NO: 18), YSRVGRSVTD (SEQ ID NO: 19), KRLKSMTD (SEQ ID NO:26), and LINCKRLKSMTD (SEQ ID NO:27) and
(b) a second domain, attached to the first domain, wherein the second domain comprises a cell-penetrating, membrane-tethering hydrophobic moiety comprising a lipid, a cholesterol, a phospholipid, a steroid, a sphingosine, a ceramide, an octylglycine, a 2- cyclohexylalanine, or a benzolylphenylalanine;
wherein said chimeric polypeptide is an antagonist of a chemokine receptor.
27. The chimeric polypeptide of claim 26 , wherein the chemokine receptor is a CXC chemokine receptor or a CC chemokine receptor.
28. The chimeric polypeptide of claim 26 , wherein said second domain comprises a lipid.
29. The chimeric polypeptide of claim 26 , wherein said second domain comprises a hydrophobic moiety that is selected from the group consisting of a: nonanoyl (C 9 ); capryl (C 10 ); undecanoyl (C 11 ); lauroyl (C 12 ); tridecanoyl (C 13 ); myristoyl (C 14 ); pentadecanoyl (C 15 ); palmitoyl (C 16 ); phytanoyl (methyl substituted C 16 ); heptadecanoyl (C 17 ); stearoyl (C 18 ); nonadecanoyl (C 19 ); arachidoyl (C 20 ); heniecosanoyl (C 21 ); behenoyl (C 22 ); trucisanoyl (C 23 ); and lignoceroyl (C 24 ) moiety.
30. The chimeric polypeptide of claim 26 , wherein the second domain is attached to the first domain with an amide bond, a sulfhydryl, an amine, an alcohol, a phenolic group, or a carbon-carbon bond.
31. The chimeric polypeptide of claim 26 , wherein the second domain comprises a palmitoyl moiety.
32. The chimeric polypeptide of claim 26 , wherein the second domain comprises a myristoyl (C 14 ) or pentadecanoyl (C 15 ) moiety.
33. The chimeric polypeptide of claim 26 , wherein the second domain comprises a lithocholic acid or a salt thereof.
34. The chimeric polypeptide of claim 26 , wherein the second domain comprises a hydrophobic moiety that is selected from the group consisting of a cholesterol, a phospholipid, a steroid, a sphingosine, a ceramide, an octyl-glycine, a 2- cyclohexylalanine, a benzolylphenylalanine, and a C 3 -C 8 fatty acid.
35. The chimeric polypeptide of claim 26 , wherein the second domain comprises a steroid.
36. The chimeric polypeptide of claim 26 , wherein the second domain comprises a hydrophobic moiety that is selected from the group consisting of a phospholipid, a steroid, a sphingosine, a ceramide, an octylglycine, a 2-cyclohexylalanine, and a benzolylphenylalanine.
37. A pharmaceutical composition comprising the chimeric peptide of claim 26 and a pharmaceutically acceptable carrier.