Thrombin peptide derivative dimers
Disclosed are thrombin peptide derivative dimers comprising two polypeptides having the amino acid sequence Arg-Gly-Asp-Ala-Cys-Glu-Gly-Asp-Ser-Gly-Gly-Pro-Phe-Val (SEQ ID NO:2), or a C-terminal truncated fragment of the polypeptide having at least six amino acids. Zero, one, two, or three amino acids in the polypeptide or polypeptide fragment differ from the corresponding position of SEQ ID NO:2. Also disclosed are methods of treating a subject in need of treatment with a thrombin receptor agonist. The methods comprise the step of administering an effective amount of the thrombin peptide derivative described above.
1. A composition comprising a peptide dimer, said dimer comprising two thrombin peptide derivatives 12 to 23 amino acids in length, which, independently, comprise the polypeptide Asp-Ala-Cys-Glu-Gly-Asp-Ser-Gly-Gly-Pro-Phe-Val (SEQ ID NO:12); said thrombin peptide derivatives optionally comprising a C-terminal amide; and said thrombin peptide derivatives optionally comprising an acylated N-terminus, wherein the composition is greater than 95% free of monomer by weight.
2. The composition of claim 1 , wherein the thrombin peptide derivatives are the same.
3. The composition of claim 2 , wherein the thrombin peptide derivatives are covalently linked through a disulfide bond.
4. The composition of claim 3 , wherein the thrombin peptide derivatives comprise a C-terminal amide and optionally comprise an acylated N-terminus, wherein said C-terminal amide is represented by —C(O)NR b R c , wherein R b and R c are independently hydrogen, a C 1 -C 10 substituted or unsubstituted aliphatic group, and said N-terminal acyl group is represented by R d C(O)—, wherein R d is hydrogen, a C 1 -C 10 substituted or unsubstituted aliphatic group.
5. The composition of claim 3 , wherein the thrombin peptide derivatives comprise a C-terminal amide and optionally comprise an acylated N-terminus, wherein said C-terminal amide is represented by —C(O)NR b R c , wherein R b and R c are independently hydrogen, a C 1 -C 10 substituted or unsubstituted aliphatic group; and said N-terminal acyl group is represented by R d C(O)—, wherein R d is hydrogen, a C 1 -C 10 substituted or unsubstituted aliphatic group.
6. The composition of claim 3 , wherein the thrombin peptide derivatives do not comprise an N-terminal acyl group and optionally comprise a C-terminal amide represented by —C(O)NH 2 .
7. The composition of claim 6 , wherein the thrombin peptide derivatives comprise the polypeptide Arg-Gly-Asp-Ala-Cys-Glu-Gly-Asp-Ser-Gly-Gly-Pro-Phe-Val (SEQ ID NO:2).
8. The composition of claim 6 , wherein the thrombin peptide derivatives comprise the amino acid sequence Ala-Gly-Tyr-Lys-Pro-Asp-Glu-Gly-Lys-Arg-Gly-Asp-Ala-Cys-Glu-Gly-Asp-Ser-Gly-Gly-Pro-Phe-Val (SEQ ID NO:5), or a fragment thereof comprising SEQ ID NO:12.
9. The composition of claim 6 , wherein the thrombin peptide derivatives have the amino acid sequence Ala-Gly-Tyr-Lys-Pro-Asp-Glu-Gly-Lys-Arg-Gly-Asp-Ala-Cys-Glu-Gly-Asp-Ser-Gly-Gly-Pro-Phe-Val (SEQ ID NO:5).
10. A composition comprising two thrombin peptide derivatives 9 to 23 amino acids in length, which independently, comprise the polypeptide Arg-Gly-Asp-Ala-Cys-X 1 -Gly-Asp-Ser-Gly-Gly-Pro-X 2 -Val (SEQ ID NO:4), wherein X 1 is Glu or Gln and X 2 is Phe, Met, Leu, His or Val); wherein the thrombin peptide derivatives are the same and are covalently linked through a disulfide bond; and wherein the thrombin peptide derivatives do not comprise an N-terminal acyl group and optionally comprise a C-terminal amide represented by —C(O)NH 2 ; and wherein the composition is greater than 95% free of monomer by weight.
11. The composition of claim 10 , wherein the thrombin peptide derivatives have the amino acid sequence Ala-Gly-Tyr-Lys-Pro-Asp-Glu-Gly-Lys-Arg-Gly-Asp-Ala-Cys-X 1 -Gly-Asp-Ser-Gly-Gly-Pro-X 2 -Val (SEQ ID NO:6), wherein X 1 is Glu or Gln and X 2 is Phe, Met, Leu, His or Val; or wherein the thrombin peptide derivatives comprise SEQ ID NO:12.
12. The composition of claim 10 , wherein the thrombin peptide derivatives have the amino acid sequence Ala-Gly-Tyr-Lys-Pro-Asp-Glu-Gly-Lys-Arg-Gly-Asp-Ala-Cys-X 1 -Gly-Asp-Ser-Gly-Gly-Pro-X 2 -Val (SEQ ID NO:6), wherein X 1 is Glu or Gln and X 2 is Phe, Met, Leu, His or Val.
13. The composition of claim 12 , wherein X 1 is Glu and X 2 is Phe.
14. The composition of claim 10 , wherein the thrombin peptide derivatives are the polypeptide H-Ala-Gly-Tyr-Lys-Pro-Asp-Glu-Gly-Lys-Arg-Gly-Asp-Ala-Cys-X 1 -Gly-Asp-Ser-Gly-Gly-Pro-X 2 -Val-NH 2 (SEQ ID NO:11), wherein X 1 is Glu or Gln and X 2 is Phe, Met, Leu, His or Val.
15. The composition of claim 14 , wherein X 1 is Glu and X 2 is Phe.
16. A composition comprising a peptide dimer having two thrombin peptide derivatives, each with the amino acid sequence Ala-Gly-Tyr-Lys-Pro-Asp-Glu-Gly-Lys-Arg-Gly-Asp-Ala-Cys-Glu-Gly-Asp-Ser-Gly-Gly-Pro-Phe-Val (SEQ ID NO:3), wherein the thrombin peptide derivatives are covalently linked by a disulfide bond; said thrombin peptide derivatives optionally comprising a C-terminal amide; and said thrombin peptide derivatives optionally comprising an acylated N-terminus; wherein the composition is greater than 95% free of monomer by weight.
17. A composition comprising a peptide dimer represented by the following structural formula:
wherein the composition is greater than 95% free of monomer by weight.
18. A composition comprising two thrombin peptide derivatives, which independently, comprise the polypeptide Ala-Gly-Tyr-Lys-Pro-Asp-Glu-Gly-Lys-Arg-Gly-Asp-Ala-Cys-Glu-Gly-Asp-Ser-Gly-Gly-Pro-Phe-Val-Met-Lys-Ser-Pro-Phe-Asn-Asn-Arg-Trp-Tyr (SEQ ID NO:7); wherein the thrombin peptide derivatives are the same and are covalently linked through a disulfide bond; and wherein the thrombin peptide derivatives do not comprise an N-terminal acyl group and optionally comprise a C-terminal amide represented by —C(O)NH 2 ; and wherein the composition is greater than 95% free of monomer by weight.