Silk Fibroin Hydrogels and Uses Thereof

• US 20110008436A1
• Filed: 04/20/2010
• Published: 01/13/2011
• Est. Priority Date: 04/20/2009
• Status: Abandoned Application

• Current Assignee: Allergan Incorporated (Abbvie Incorporated)
• Original Assignee: Allergan Incorporated (Abbvie Incorporated)
• Inventors: COLLETTE, ADAM L., CHEN, JINGSONG, ALTMAN, GREGORY H., HORAN, REBECCA L.
• Application Number:
  US12/764,038
• Publication Number:
  US 20110008436A1
• Time in Patent Office: Days
• US Class Current: 424/484
• CPC Class Codes: A61F 2/12 Mammary prostheses and implants
• CPC Class Codes: A61K 2800/5922 At least two compounds being classified in the same subclass of A61K8/18
• CPC Class Codes: A61K 2800/91 Injection
• CPC Class Codes: A61K 38/1767 from invertebrates
• 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: A61K 8/64 Proteins; Peptides; Derivatives or degradation products thereof
• CPC Class Codes: A61L 2430/34 for soft tissue reconstruction
• CPC Class Codes: A61L 27/227 Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
• CPC Class Codes: A61L 27/26 Mixtures of macromolecular compounds
• CPC Class Codes: A61L 27/52 Hydrogels or hydrocolloids
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 23/02 Local anaesthetics
• CPC Class Codes: A61P 41/00 Drugs used in surgical methods, e.g. surgery adjuvants for preventing adhesion or for vitreum substitution
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/06 Antiarrhythmics
• CPC Class Codes: A61Q 19/00 Preparations for care of the skin
• CPC Class Codes: C08B 37/0072 Hyaluronic acid, i.e. HA or hyaluronan; Derivatives thereof, e.g. crosslinked hyaluronic acid (hylan) or hyaluronates
• CPC Class Codes: C08L 89/00 Compositions of proteins; Compositions of derivatives thereof foodstuff preparations A23J3/00

Abstract

The present specification provides for methods for purifying fibroins, purified fibroins, methods of conjugating biological and synthetic molecules to fibroins, fibroins conjugated to such molecules, methods of making fibroin hydrogels, fibroin hydrogels and fibroin hydrogel formulations useful for a variety of medical uses, including, without limitation uses as bulking agents, tissue space fillers, templates for tissue reconstruction or regeneration, cell culture scaffolds for tissue engineering and for disease models, surface coating to improve medical device function, or drug delivery devices.

Claims

1. A hydrogel formulation comprising:

a) a gel phase, the gel phase including hydrogel particles comprising a substantially sericin-depleted silk fibroin and an amphiphilic peptide; and

b) a carrier phase.

2. The hydrogel formulation of claim 1, wherein the hydrogel particles comprises about 1% (w/v) to about 10% (w/v) of silk fibroin.

3. The hydrogel formulation of claim 1, wherein the amphiphilic peptide comprising a RGD motif.

4. The hydrogel formulation of claim 3, wherein the amphiphilic peptide is 23 RGD.

5. The hydrogel formulation of claim 1, wherein the amphiphilic peptide comprises of a tail region, followed by a spacer region and finally a RGD motif.

6. The hydrogel formulation of claim 1, wherein the hydrogel particles comprises a molar ratio of 1:

10 to 10;

1 moles of the amphiphilic peptide per mole of the silk fibroin.

7. The hydrogel formulation of claim 6, wherein the hydrogel particles comprises a molar ratio of 3:

1 moles of the amphiphilic peptide per mole of the silk fibroin.

8. The hydrogel formulation of claim 1, wherein the hydrogel particles comprises a protein structure having a β

-sheet conformation of at least 80%.

9. The hydrogel formulation of claim 1, wherein the hydrogel particles comprises a protein structure having a β

-sheet conformation of at least 50%.

10. The hydrogel formulation of claim 1, wherein the hydrogel particles comprises a protein structure having a β

-sheet conformation of at least 20%.

11. The hydrogel formulation of claim 1, wherein the hydrogel particles comprises a protein structure having an α

-helical and random coil conformation of at most 20%.

12. The hydrogel formulation of claim 1, wherein the hydrogel particles have a cross-sectional area from about 5 μ

m² to about 1000 μ

m².

13. The hydrogel formulation of claim 1, wherein the hydrogel particles have a cross-sectional area from about 20 μ

m² to about 50 μ

m².

14. The hydrogel formulation of claim 1, wherein the carrier phase comprises saline.

15. The hydrogel formulation of claim 1, wherein the carrier phase comprises a surfactant solution.

16. The hydrogel formulation of claim 1 wherein the gel phase is 50% to 99% of the total formulation volume, the remainder being a carrier solution.

17. The hydrogel formulation of claim 16, wherein the gel phase is 75% of the total formulation volume, the remainder being a carrier solution.

18. The hydrogel formulation of claim 1, wherein the hydrogel formulation further comprising lidocaine.

19. The hydrogel formulation of claim 1, wherein a synthetic molecule having the formula:

(molecule X)_n-(spacer peptide)_0-300-(five-amino-acid peptide tail) is conjugated to the silk fibroin.

20. The hydrogel formulation of claim 1, wherein, upon injection, the hydrogel particles remains substantially at the injection site for one month to eighteen months.

21. The hydrogel formulation of claim 4, wherein the said formulation, upon injection, remains substantially at the injection site for five weeks to six weeks and consists of about 1% (w/v) to about 3% (w/v) silk fibroin, about 20% (v/v) to about 50% (v/v) saline, an average particle size range of about 20 μ

m² to about 50 μ

m².

22. The hydrogel formulation of claim 4, wherein the said formulation, upon injection, remains substantially at the injection site for two months to three months and consists of about 3% (w/v) to about 5% (w/v) silk fibroin, about 20% (v/v) to about 50% (v/v) saline, an average particle size range of about 20 μ

m² to about 40 μ

m².

23. The hydrogel formulation of claim 4, wherein the said formulation, upon injection, remains substantially at the injection site for five months to six months and consists of about 4% (w/v) to about 6% (w/v) silk fibroin, about 20% (v/v) to about 50% (v/v) saline, an average particle size range of about 20 μ

m² to about 40 μ

m².

24. The hydrogel formulation of claim 4, wherein the said formulation, upon injection, remains substantially at the injection site for eight months to ten months and consists of about 6% (w/v) to about 8% (w/v) silk fibroin, about 20% (v/v) to about 40% (v/v) saline, an average particle size range of about 20 μ

m² to about 40 μ

m².

25. The hydrogel formulation of claim 4, wherein the said formulation, upon injection, remains substantially at the injection site for eight months to ten months and consists of about 6% (w/v) to about 8% (w/v) silk fibroin, about 20% (v/v) to about 40% (v/v) saline, an average particle size range of about 20 μ

m² to about 40 μ

m².

26. The hydrogel formulation of claim 4, wherein the said formulation, upon injection, remains substantially at the injection site for twelve months to eighteen months and consists of about 8% (w/v) to about 10% (w/v) silk fibroin, about 0% (v/v) to about 40% (v/v) saline, an average particle size range of about 20 μ

m² to about 1000 μ

m².

27. A method of making a hydrogel formulation, the method comprising the steps of:

a) obtaining a solution comprising a substantially sericin-free, depolymerized silk fibroin; and

b) adding an enhancer solution comprising an alcohol and an amphiphilic peptide to the silk fibroin solution;

c) rinsing the hydrogel in a solvent to remove residual enhancer solution

d) homogenizing hydrogel in a physiological-acceptable buffer to form hydrogel particles;

28. The method of claim 27, wherein the method further comprises step (e) milling the hydrogel particles to a desired particle size.