Hydroxyphenyl cross-linked macromolecular network and applications thereof

• US 6,982,298 B2
• Filed: 01/08/2004
• Issued: 01/03/2006
• Est. Priority Date: 01/10/2003
• Status: Active Grant

• Current Assignee: Cleveland Clinic Foundation
• Original Assignee: Cleveland Clinic Foundation
• Inventors: Gross, Richard A., Darr, Aniq B., Calabro, Anthony
• Primary Examiner(s): Seidleck, James J.
• Assistant Examiner(s): Zemel, Irina S.
• Application Number:
  US10/753,779
• Publication Number:
  US 20040147673A1
• Time in Patent Office: 726 Days
• Field of Search: 525/54.1, 525/54.2, 525/420, 525/326, 525/540, 527/600
• US Class Current: 525/54.1
• CPC Class Codes: A61K 47/34 Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers A61K47/10 takes precedence
• CPC Class Codes: A61K 47/36 Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
• CPC Class Codes: A61K 47/42 Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein oligopeptides having up to five amino acids A61K47/183; polyamino acids A61K47/34
• CPC Class Codes: A61L 27/14 Macromolecular materials
• CPC Class Codes: A61L 27/52 Hydrogels or hydrocolloids
• CPC Class Codes: C08G 63/81 using solvents C08G63/79 takes precedence
• CPC Class Codes: C08G 73/02 Polyamines
• CPC Class Codes: C08J 3/075 Macromolecular gels
• CPC Class Codes: C08L 101/14 the macromolecular compounds being water soluble or water swellable, e.g. aqueous gels

Abstract

A dihydroxyphenyl cross-linked macromolecular network is provided that is useful in artificial tissue and tissue engineering applications, such as artificial or synthetic cartilage. The network is made by first providing a polyamine or polycarboxylate macromolecule (having a plurality of amine or carboxylic acid groups respectively attached along the length of the molecule), reacting this macromolecule with a hydroxyphenyl compound having a free carboxylic acid group in the case of a polyamine or a free primary amine group in the case of a polycarboxylate, and substituting the hydroxyphenyl compound onto the macromolecule via a carbodiimide-mediated reaction pathway to provide a hydroxyphenyl-substituted macromolecule. This macromolecule is then linked to other such macromolecules via an enzyme catalyzed dimerization reaction between two hydroxyphenyl groups attached respectively to different macromolecules under metabolic conditions of temperature and pH. In a preferred embodiment, the macromolecular network is made up of tyramine-substituted hyaluronan molecules that are linked by dityramine bonds to provide a stable, coherent hydrogel with desired physical properties. A method of preparing such a network is also provided.

Claims

1. A macromolecular network comprising

wherein R₁ and R₂ each comprises a structure selected from the group consisting of polycarboxylates, polyamines, polyhydroxyphenyl molecules, and copolymers thereof, and wherein R₁ and R₂ can be the same or different structures.

2. A macromolecular network according to claim 1, wherein R₁ is a polycarboxylate.

3. A macromolecular network according to claim 1, wherein R₁ is a polyamine.

4. A macromolecular network according to claim 1, wherein R₁ is a polyphenol.

5. A macromolecular network according to claim 1, wherein R₁ comprises a structure selected from the group consisting of peptides and proteins.

6. A macromolecular network according to claim 1, wherein R₁ comprises a structure selected from the group consisting of glycosaminoglycans.

7. A macromolecular network according to claim 1, wherein R₁ comprises hyaluronan.

8. A macromolecular network according to claim 1, wherein R₁ comprises a structure selected from the group consisting of chondroitin sulfate and dermatan sulfate.

9. A macromolecular network according to claim 1, wherein R₁ comprises a structure selected from the group consisting of heparan sulfate and heparin.

10. A macromolecular network according to claim 1, wherein R₁ comprises a structure selected from the group consisting of alginate, polyglucuronic acid, polygalacturonic acid, pectin and colominic acid.

11. A macromolecular network according to claim 1, wherein R₁ comprises a structure selected from the group consisting of polyaspartic acid and polyglutamic acid.

12. A macromolecular network according to claim 1, wherein R₁ comprises polytyrosine.

13. A macromolecular network according to claim 1, wherein R₁ comprises a structure selected from the group consisting of polylysine and polyarginine.

14. A macromolecular network according to claim 1, wherein R₁ comprises a proteoglycan.

15. A macromolecular network according to claim 1, wherein R1 comprises aggrecan.

16. A macromolecular network according to claim 1, further comprising a population of viable living cells within the macromolecular network.

17. A macromolecular network according to claim 1, further comprising bioactive factors within the macromolecular network.

18. A macromolecular network according to claim 1, said network comprising polycarboxylate molecules that have been substituted with a hydroxyphenyl compound, wherein at least one dihydroxyphenyl linkage is formed between two hydroxyphenyl groups attached respectively to adjacent polycarboxylate molecules.

19. A macromolecular network according to claim 1, said network comprising polyamine molecules that have been substituted with a hydroxyphenyl compound, wherein at least one dihydroxyphenyl linkage is formed between two hydroxyphenyl groups attached respectively to adjacent polyamine molecules.

20. A macromolecular network according to claim 1, said network comprising peptides and/or proteins that have been substituted with a hydroxyphenyl compound, wherein at least one dihydroxyphenyl linkage is formed between two hydroxyphenyl groups attached respectively to adjacent peptides and/or proteins.

21. A macromolecular network according to claim 18, said hydroxyphenyl compound being tyramine or tyrosine.

22. A macromolecular network according to claim 19, said hydroxyphenyl compound being tyrosine, 4-hydroxyphenylacetic acid, or 3-(4-hydroxyphenyl)propionic acid.

23. A macromolecular network according to claim 18, said polycarboxylate molecules having a hydroxyphenyl compound substitution rate less than 50 percent based on the molar quantity of CO₂H sites present on said polycarboxylate molecules.

24. A macromolecular network according to claim 19, said polyamine molecules having a hydroxyphenyl compound substitution rate less than 50 percent based on the molar quantity of primary NH₂ sites present on said polyamine molecules.

25. A macromolecular network according to claim 1, comprising a plurality of tyramine-substituted hyaluronan molecules, at least two adjacent hyaluronan molecules being linked by a dityramine linkage.

26. A macromolecular network according to claim 25, having a tyramine substitution rate on said hyaluronan molecules of less than 50% based on the molar quantity of CO₂H sites present on said hyaluronan molecules.

27. A macromolecular network according to claim 25, having a tyramine substitution rate on said hyaluronan molecules of less than 10% based on the molar quantity of CO₂H sites present on said hyaluronan molecules.

28. A macromolecular network according to claim 25, further comprising a population of viable living cells within the macromolecular network.

29. A macromolecular network according to claim 25, further comprising bioactive factors within the macromolecular network.

30. A hydrogel comprising the macromolecular network according to claim 25.

31. A hydrogel according to claim 30, having rigidity and rheological properties of a jelly.

32. A hydrogel according to claim 30, having rigidity and rheological properties of a gelatin.

33. A hydrogel according to claim 30, having rigidity and rheological properties of a dough.

34. A hydrogel according to claim 30, having rigidity and rheological properties of a resilient rubber-like composition.

35. A hydrogel according to claim 30, having cartilage-like rigidity and rheological properties.

36. A hydrogel according to claim 30, further comprising a population of viable living cells within the macromolecular network.

37. A hydrogel according to claim 30, provided as synthetic or artificial cartilage.

38. A hydrogel according to claim 30, further comprising bioactive factors within the macromolecular network.

39. A macromolecular network according to claim 28, said population of viable living cells including cells selected from the group consisting of chondrocytes, progenitor cells and stem cells.

40. A hydrogel according to claim 36, said population of viable living cells including cells selected from the group consisting of chondrocytes, progenitor cells and stem cells.

41. A macromolecular network according to claim 29, said bioactive factors including factors selected from the group consisting of growth factors, hormones and factors controlling cell differentiation.

42. A hydrogel according to claim 38, said bioactive factors including factors selected from the group consisting of growth factors, hormones and factors controlling cell differentiation.

43. A macromolecular network according to claim 25, having a tyramine substitution rate on said hyaluronan molecules of 4–

6 percent based on the molar quantity of CO₂H sites present on said hyaluronan molecules.

44. A hydrogel according to claim 30, said macromolecular network having a tyramine substitution rate on said hyaluronan molecules of 4–

6 percent based on the molar quantity of CO₂H sites present on said hyaluronan molecules.

45. A macromolecular network according to claim 25, having a tyramine substitution rate on said hyaluronan molecules of 1.7 percent based on the molar quantity of CO₂H sites present on said hyaluronan molecules.

46. A hydrogel according to claim 30, said macromolecular network having a tyramine substitution rate on said hyaluronan molecules of 1.7 percent based on the molar quantity of CO₂H sites present on said hyaluronan molecules.

47. A macromolecular network according to claim 25, having a tyramine substitution rate on said hyaluronan molecules of 4.7 percent based on the molar quantity of CO₂H sites present on said hyaluronan molecules.

48. A hydrogel according to claim 30, said macromolecular network having a tyramine substitution rate on said hyaluronan molecules of 4.7 percent based on the molar quantity of CO₂H sites present on said hyaluronan molecules.