Cartilage particle tissue mixtures optionally combined with a cancellous construct

• US 20100274362A1
• Filed: 01/14/2010
• Published: 10/28/2010
• Est. Priority Date: 01/15/2009
• Status: Abandoned Application

• Current Assignee: Musculoskeletal Transplant Foundation, ProChon Biotech Ltd. (Ocugen, Inc.)
• Original Assignee: Musculoskeletal Transplant Foundation
• Inventors: Barkay-Olami, Hilla, Stern, Baruch, Munson, John C., Yayon, Avner, Semler, Eric J., Huang, Yen-Chen, Yannariello-Brown, Judith I., Tomford, William W., Jacobs, Morris L., Sunwoo, Moon Hae, Truncale, Katherine G., Gertzman, Arthur A., Callahan, Alex B., Shikhanovich, Roman
• Application Number:
  US12/657,207
• Publication Number:
  US 20100274362A1
• Time in Patent Office: Days
• US Class Current: 623/23.720
• CPC Class Codes: A61K 35/32 Bones; Osteocytes; Osteoblasts; Tendons; Tenocytes; Teeth; Odontoblasts; Cartilage; Chondrocytes; Synovial membrane
• CPC Class Codes: A61K 38/1825 Fibroblast growth factor [FGF]
• CPC Class Codes: A61K 9/19 lyophilised , i.e. freeze-dried, solutions or dispersions lyophilised products with subsequent particle size reduction A61K9/14; granules or pellets made by lyphilisation A61K9/1682; solid oral dosage forms made by lyophilisation A61K9/2095; lyophilisation additives A61K47/00
• CPC Class Codes: A61L 2300/414 Growth factors
• CPC Class Codes: A61L 2300/602 Type of release, e.g. controlled, sustained, slow
• CPC Class Codes: A61L 27/3612 Cartilage, synovial fluid
• CPC Class Codes: A61L 27/3654 Cartilage, e.g. meniscus
• CPC Class Codes: A61L 27/54 Biologically active materials, e.g. therapeutic substances A61L27/227 takes precedence
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders

Abstract

Mixtures, such as gels or pastes, comprising freeze-milled cartilage particles and exogenous growth factors are used for repairing chondral defects. Such mixtures may be applied to constructs comprising cancellous bone for implantation at the defect site. Suitable growth factors include variants of FGF-2, particularly variants that include a sole amino acid substitution for asparagine at amino acid 111 of the β8-β9 loop of the FGF-2 peptide. Such FGF-2 variants are released slowly and continuously at a constant rate from cartilage pastes. In other embodiments, the amino acid substituted for asparigine is glycine. Other variants that may be used include FGF-9 variants having truncated chains and a sole amino acid substitution in the β8-β9 loop of the FGF-9 peptide either for tryptophan at amino acid 144 or for asparagine at amino acid 143.

Claims

1. A sustained release composition, comprising lyophilized freeze-milled cartilage particles having a size no greater than 1 mm and an active growth factor exogenous to said cartilage particles, said growth factor being incorporated into said composition such that said growth factor is released substantially continuously over a prolonged period of time.

2. The composition of claim 1, further comprising a biocompatible carrier.

3. The composition of claim 2, wherein said biocompatible carrier is biodegradable.

4. The composition of claim 2, wherein said biocompatible carrier includes freeze-dried fibrin.

5. The composition of claim 4, wherein said fibrin is in the form of a matrix having open pores.

6. The composition of claim 4, wherein said fibrin is selected from the group consisting of human plasma protein, non-human mammalian plasma protein, avian plasma protein, recombinant plasma protein, partially-purified plasma protein, and totally purified plasma protein.

7. The composition of claim 1, wherein said cartilage particles have a size less than 210 microns.

8. The composition of claim 7, wherein said cartilage particles have a size in the range of from about 10 microns to about 210 microns, and constitute from about 10% to about 80% by weight of the composition.

9. The composition of claim 1, wherein said cartilage particles are selected from the group consisting of allograft cartilage, autograft cartilage and xenograft cartilage.

10. The composition of claim 9, wherein said cartilage particles are allograft cartilage.

11. The composition of claim 1, wherein said prolonged period of time is in the range of at least 2 days to about 10 days.

12. The composition of claim 11, wherein said growth factor is reversibly bound to said cartilage particles such that said growth factor is released substantially continuously into an aqueous medium in vitro over said prolonged period of time.

13. The composition of claim 1, wherein said prolonged period of time is in the range of at least 10 days to about 100 days.

14. The composition of claim 13, wherein said growth factor is reversibly bound to said cartilage particles such that said growth factor is released substantially continuously into an aqueous medium in vitro over said prolonged period of time.

15. The composition of claim 1, wherein said prolonged period of time is in the range of at least 2 days to about 120 days.

16. The composition of claim 15, wherein said growth factor is reversibly bound to said cartilage particles such that said growth factor is released substantially continuously into an aqueous medium in vitro over said prolonged period of time.

17. The composition of claim 1, wherein said prolonged period of time in the range of at least 2 days to about 180 days.

18. The composition of claim 17, wherein said growth factor is reversibly bound to said cartilage particles such that said growth factor is released substantially continuously into an aqueous medium in vitro over said prolonged period of time.

19. The composition of claim 1, wherein said growth factor is released into an aqueous medium in vitro such that the concentration of said growth factor in said aqueous medium varies by a factor of less than about 10 over said prolonged period of time.

20. The composition of claim 1, wherein said composition has an initial concentration of said exogenous growth factor in the range of about 0.5 μ

g per ml of said composition to about 5,000 μ

g per ml of said composition.

21. The composition of claim 1, wherein said growth factor is selected from the group consisting of native, recombinant and variant isoforms of FGF-2, FGF-4, FGF-5, FGF-7, FGF-9, FGF-11, FGF-21, TGF-β

, BMP-2, BMP-4, BMP-7, BMP-14, PDGF, VEGF, and IGF-1.

22. The composition of claim 1, wherein said growth factor is selected from the group consisting of FGF isoforms having a least one substitution of an amino acid in the β

8-β

9 loop.

23. The composition of claim 22, wherein said at least one substitution is a single point substitution.

24. The composition of claim 1, wherein said growth factor is selected from the group consisting of native, recombinant and variant isoforms of FGF-2.

25. The composition of claim 1, wherein said growth factor is a human FGF-2.

26. The composition of claim 1, wherein said growth factor is a variant of a native human FGF-2, said variant having a substitution of amino acid 111 from asparagine to an amino acid selected from the group consisting of glycine, arginine, valine, glutamic acid and serine and is SEQ ID NO:

1.

27. The composition of claim 1, wherein said growth factor is a variant of a native human FGF-2, said variant having a substitution of amino acid 111 from asparagine to glycine and is SEQ ID NO:

3.

28. The composition of claim 1, wherein said growth factor is a variant of a native human FGF-2, said variant having a substitution of amino acid 3 from alanine to glutamine, a substitution of amino acid 5 from serine to glutamine, and a substitution of amino acid 111 from asparagine to an amino acid selected from the group consisting of glycine, arginine, valine, glutamic acid and serine and is SEQ ID NO:

4.

29. The composition of claim 1, wherein said growth factor is a variant of a native human FGF-2, said variant having a substitution of amino acid 3 from alanine to glutamine, a substitution of amino acid 5 from serine to glutamine, and a substitution of amino acid 111 from asparagine to glycine and is SEQ ID NO:

5.

30. The composition of claim 1, wherein said growth factor is selected from the group consisting of native, recombinant and variant isoforms of FGF-9.

31. The composition of claim 1, wherein said growth factor is a human FGF-9.

32. The composition of claim 1, wherein said growth factor is a variant of a native human FGF-9, said variant having a 63 amino acid N-terminus truncation and a substitution of amino acid 144 from tryptophan to an amino acid selected from the group consisting of glycine, arginine, glutamic acid and valine, and is SEQ ID NO:

6.

33. The composition of claim 1, wherein said growth factor is a variant of a native human FGF-9, said variant having a 63 amino acid N-terminus truncation, an 18 amino acid C-terminus truncation, and a substitution of amino acid 144 from tryptophan to an amino acid selected from the group consisting of glycine, arginine, glutamic acid and valine, and is SEQ ID NO:

7.

34. The composition of claim 1, wherein said growth factor is a variant of a native human FGF-9, said variant having a 63 amino acid N-terminus truncation and a substitution of amino acid 143 from asparagine to serine, and is SEQ ID NO:

8.

35. The composition of claim 1, wherein said growth factor is a variant of a native human FGF-9, said variant having a 63 amino acid N-terminus truncation, an 18 amino acid C-terminus truncation, and a substitution of amino acid 143 from asparagine to serine, and is SEQ ID NO:

9.

36. The composition of claim 4, wherein said fibrin is obtained by mixing plasma proteins comprising fibrinogen and Factor XIII with thrombin, and wherein said composition includes at least one anti-fibrinolytic agent.

37. The composition of claim 2, wherein said biocompatible carrier comprises a carrier selected from the group consisting of phosphate buffered saline solution, saline solution, sodium hyaluronate solution, hyaluronic acid and its derivatives, gelatin, collagen, chitosan, alginate, dextran, carboxymethylcellulose, and hydroxypropyl methylcellulose.

38. The composition of claim 37, wherein said biocompatible carrier is selected from the group consisting of sodium hyaluronate solution, hyaluronic acid and its derivatives and phosphate buffered saline solution.

39. A cartilage repair construct, comprising a demineralized bone portion which contains the composition of claim 1.

40. A method of repairing a defect in cartilage in a subject in need thereof, comprising the step of placing, in a defect site, a sustained release composition including lyophilized freeze-milled cartilage particles having a size no greater than 1 mm and an active growth factor exogenous to said cartilage particles, said growth factor being incorporated into said composition such that said growth factor is released substantially continuously over a prolonged period of time.

41. The method of claim 40, wherein the defect site is in articular cartilage.

42. The method of claim 40, wherein said step of placing the sustained release composition in a defect site at least partially relieves the need for a prosthetic replacement of a body part in which the defect site is located.

43. The method of claim 40, wherein said growth factor is reversibly bound to said cartilage particles such that said growth factor is released substantially continuously over a period of time.

44. A composition, comprising lyophilized freeze-milled cartilage particles having a size no greater than 1 mm and a biocompatible carrier including freeze-dried fibrin.