Collagen peptide conjugates and uses therefor
Described herein are conjugates of collagen peptides and metal binding agents and compositions resulting therefrom, useful in various tissue engineering and regeneration applications, in cell culture, cell adhesion, cosmetic surgery, construction of artificial skin substitutes, management of severe burns and burn surgery, reconstruction of bone and a wide variety of dental, orthopedic and surgical purposes, as drug delivery vehicles and in delivering populations of cells to a site of disease or injury.
1. A synthetic collagen conjugate capable of forming a type II helix, the conjugate comprising one or more metal-binding moieties, and a peptide comprising a plurality of tripeptides, each of which comprises proline or hydroxyproline, or a combination thereof; where the one or more metal-binding moieties are covalently attached to the peptide, optionally with a divalent linker;
wherein one of said metal-binding moieties is covalently attached to the N-terminus of the peptide, and one of said metal-binding moieties is covalently attached to the C-terminus of the peptide; or
wherein one of said one or more metal-binding moieties is covalently attached to a non-terminal amino acid of the peptide; or
wherein one of said metal-binding moieties is covalently attached to the N-terminus of the peptide, one of said metal-binding moieties is covalently attached to the C-terminus of the peptide, and one of said metal-binding moieties is covalently attached to a non-terminal amino acid of the peptide; and
wherein the conjugate may further comprise a drug or a diagnostic or imaging agent, where the drug or the diagnostic or imaging agent is covalently attached to the peptide, optionally with a divalent linker.
2. The conjugate of claim 1 wherein the peptide comprises a plurality of tripeptides, each of which comprises glycine and proline.
3. The conjugate of claim 1 wherein the peptide comprises a plurality of tripeptides, each of which comprises glycine and hydroxyproline.
4. The conjugate of claim 1 wherein one of said metal-binding moieties is covalently attached to the N-terminus of the peptide, and one of said metal-binding moieties is covalently attached to the C-terminus of the peptide.
5. The conjugate of claim 1 wherein the conjugate is capable of forming a self assembling triple helix.
6. The conjugate of claim 5 wherein the triple helix is capable of aggregating in the presence of a transition metal.
7. The conjugate of claim 1 wherein the peptide is at least about 18 amino acids in length.
8. The conjugate of claim 1 wherein the peptide is between about 18 and about 54 amino acids in length.
9. The conjugate of claim 1 wherein the peptide comprises at least about 25% glycine.
10. The conjugate of claim 1 wherein the peptide comprises at least about 5% proline.
11. The conjugate of claim 1 wherein the peptide comprises a plurality of divalent tripeptides selected from the group consisting of Xaa-Yaa-Gly, Gly-Pro-Xaa and Gly-Xaa-Hyp, or a combination thereof, where each Xaa and Yaa is independently selected in each instance from the group consisting of naturally occurring amino acids and derivatives of naturally occurring amino acids.
12. The conjugate of claim 1 wherein one or more of the metal-binding moieties is selected from the group consisting of bipyridinyls, amino bis(acetic acid)s, and His x , where x is an integer from 2 to 4, and amides thereof, and pharmaceutically acceptable salts thereof.
13. The conjugate of claim 1 wherein one or more of the metal-binding moieties is capable of binding a metal cation selected from the group consisting of cations of iron, nickel, cobalt, copper, zinc, and ruthenium, and combinations thereof.
14. The conjugate of claim 1 further comprising a drug or a diagnostic or imaging agent, where the drug or the diagnostic or imaging agent is covalently attached to the peptide, optionally with a divalent linker.
15. The conjugate of claim 14 wherein the drug is a compound capable of treating a bone or cartilage disease.
16. The conjugate of claim 14 wherein the drug or the diagnostic or imaging agent is selected from the group consisting of cell adhesion agents, growth factors, integrin binding domain peptides, REDV peptides, RGD peptides, YIGSR peptides, vascular endothelial growth factors, transforming growth factors, bone morphogenetic protein 2, epidermal growth factors, fibroblast growth factors, hepatocyte growth factors, biotin, bone antiresorptive agents, parathyroid hormone, parathyroid hormone fragments, and NBD fluorophores.
17. The conjugate of claim 1 wherein one of said one or more metal-binding moieties is covalently attached to a non-terminal amino acid of the peptide.
18. The conjugate of claim 1 wherein one of said metal-binding moieties is covalently attached to the N-terminus of the peptide, one of said metal-binding moieties is covalently attached to the C-terminus of the peptide, and one of said metal-binding moieties is covalently attached to a non-terminal amino acid of the peptide.
19. The conjugate of claim 1 having a nitrilotriacetic acid (NTA) unit at the N-terminus and a His 2 unit at the C-terminus.
20. The conjugate of claim 1 wherein an N-terminal metal-binding ligand, if present, comprises a residue having the formula
a C-terminal metal-binding ligand, if present, comprises a His-His-NH 2 residue; and
a metal-binding ligand attached to a non-terminal amino acid, if present, comprises a 4′-methyl-2,2′-bipyridine-4-carbonyl or an iminodiacetate residue.
21. The conjugate of claim 1 which is selected from the group consisting of
in which R is
or
R is an RGD-based peptide.
22. A collagen composition comprising a collagen and one or more conjugates of claim 1 .
23. A composition comprising one or more conjugates of claim 1 and one or more populations of cells.
24. The composition of claim 23 wherein the population of cells is selected from the group consisting of adipose derived stem cells (ASC), human umbilical vein endothelial cells (HUVEC), mesenchymal stem cells (MSC), and combinations thereof.