Silk-derived protein for treating inflammation
Described herein are methods for reducing inflammation by administration of an effective amount of silk-derived proteins (SDP) or a fraction thereof to a subject having an inflammatory condition. The methods include the treatment of inflammatory conditions and wounds, including corneal wounds, comprising the topical administration of an effective amount of SDP material as described herein.
1. A fibroin-derived protein composition that possesses enhanced stability in an aqueous solution compared to native silk fibroin consisting of the a fibroin-derived protein composition wherein:
the primary amino acid sequences of the fibroin-derived protein composition differ from native fibroin by at least 4% with respect to the absolute values of the combined differences in amino acid content of serine, glycine, and alanine;
cysteine disulfide bonds between the fibroin heavy and fibroin light protein chains of fibroin are reduced or eliminated;
the composition has a serine content that is reduced by greater than 25% compared to native fibroin protein, wherein the serine content is at least about 5%; and
wherein the average molecular weight of the fibroin-derived protein composition is less than 40 kDa and greater than 10 kDa, and 39%+/−10% of the protein chains of the protein composition have a molecular weight within the range of 25 kDa to 50 kDa.
2. The protein composition of claim 1 wherein the protein composition does not gel upon sonication of an aqueous solution of the protein composition at concentrations of up to 10% w/w.
3. The protein composition of claim 1 wherein the protein composition comprises less than 8% serine amino acid residues.
4. The protein composition of claim 1 wherein the protein composition comprises greater than 46.5% glycine amino acids.
5. The protein composition of claim 1 wherein the protein composition comprises greater than 30.5% alanine amino acids.
6. The protein composition of claim 1 wherein the protein composition completely re-dissolves in water after being dried to a thin film.
7. The protein composition of claim 1 wherein the protein composition lacks beta-sheet protein structure in aqueous solution.
8. The protein composition of claim 1 wherein the protein composition maintains an optical absorbance in aqueous solution of less than 0.25 at 550 nm after at least five seconds of sonication.
9. An ophthalmic formulation comprising the protein composition of claim 1 and water, and optionally one or more of a buffering medium, a salt, a stabilizer, a preservative, and a lubricant.
10. A method for reducing inflammation comprising administering the fibroin-derived protein composition according to claim 1 to inflamed tissue
thereby reducing transcription factor signaling within cell nuclei of the tissue, thereby reducing the inflammation.
11. The method of claim 10 wherein the administration to inflamed tissue reduces transcription of one or more of the inflammatory genes TNF-α, MMP-9, IL-1β, and IL-6.
12. The method of claim 10 wherein the administration is to the cornea and the administration reduces the presence of MMP-9 in the cornea.
13. The method of claim 10 wherein the administration is to the eye and the administration reduces inflammation on the ocular surface.
14. The method of claim 10 wherein the reduction in inflammation is accompanied by increased cell migration rates at the point of inflammation.
15. The method of claim 10 wherein the protein composition has an average molecular weight less than 35 kDa.
16. The method of claim 10 wherein the fibroin-derived protein composition is dissolved in an ophthalmic formulation comprising one or more of a buffering medium, a salt, a stabilizer, a preservative, and a lubricant.
17. The method of claim 10 wherein the inflammation is caused by an ocular condition, wherein the ocular condition is dry eye syndrome, corneal ulcer, corneal erosion, corneal abrasion, corneal degeneration, corneal perforation, corneal scarring, epithelial defect, keratoconjunctivitis, idiopathic uveitis, corneal transplantation, age-related macular degeneration, diabetic eye, blepharitis, glaucoma, ocular hypertension, post-operative eye pain and inflammation, posterior segment neovascularization, proliferative vitreoretinopathy, cytomegalovirus retinitis, endophthalmitis, choroidal neovascular membrane, vascular occlusive disease, allergic eye disease, tumor, retinitis pigmentosa, eye infection, scleritis, ptosis, miosis, eye pain, mydriasis, neuralgia, cicatrizing ocular surface disease, ocular infection, inflammatory ocular disease, ocular surface disease, corneal disease, retinal disease, ocular manifestations of systemic diseases, hereditary eye condition, ocular tumor, increased intraocular pressure, herpetic infection, ptyrigium or scleral tumor, wound sustained to ocular surface, post-photorefractive keratotomy eye pain and inflammation, thermal or chemical burn to the cornea, scleral wound, or keratoconus and conjunctival wound.
18. The method of claim 17 wherein the inflammation is caused by dry eye syndrome.