# US 2019/0169243 A1 · Silk-derived protein for treating inflammation

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 2019/0169243 A1
- Document ID: US20190169243A1
- Status: Published application
- Kind: A1
- Application number: 16/324,844
- Filed: 2017-08-12
- Published: 2019-06-06
- Earliest claimed: 2016-08-12
- Assignees: Silk Technologies, Ltd.
- Applicants: Silk Technologies, Ltd.
- Inventors: Brian D. LAWRENCE; David W. INFANGER
- Web page: https://patentgrep.com/patent/US20190169243A1

## CPC

- C07K 14/43586: {from silkworms}
- A61F 2/14: Eye parts, e.g. lenses or corneal implants; Artificial eyes
- A61K 9/0048: {Eye, e.g. artificial tears}
- A61K 38/012: {from animals}
- A61K 47/02: Inorganic compounds
- A61K 47/10: Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
- A61K 47/183: {Amino acids, e.g. glycine, EDTA or aspartame}
- A61K 47/26: Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
- 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)
- A61K 47/38: Cellulose; Derivatives thereof
- 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)
- A61P 27/02: Ophthalmic agents
- A61P 27/06: Antiglaucoma agents or miotics
- A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- A61F: FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS (dental prosthetics A61C)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

## Abstract

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.

## Claims

### Claim 1

A fibroin-derived protein composition that possesses enhanced stability in an aqueous solution, 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;
a plurality of peptide chains in the protein composition terminate in amide (—C(═O)NH2) groups;
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 2 kDa.

### Claim 2 (depends on 1)

The protein composition of claim 1 wherein greater than 50% of the protein chains of the protein composition have a molecular weight within the range of 10 kDa to 60 kDa.

### Claim 3 (depends on 1)

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.

### Claim 4 (depends on 1)

The protein composition of claim 1 wherein the protein composition comprises less than 8% serine amino acid residues.

### Claim 5 (depends on 1)

The protein composition of claim 1 wherein the protein composition comprises greater than 46.5% glycine amino acids.

### Claim 6 (depends on 1)

The protein composition of claim 1 wherein the protein composition comprises greater than 30.5% alanine amino acids.

### Claim 7 (depends on 1)

The protein composition of claim 1 wherein the protein composition completely re-dissolves in water after being dried to a thin film.

### Claim 8 (depends on 1)

The protein composition of claim 1 wherein the protein composition lacks beta-sheet protein structure in aqueous solution.

### Claim 9 (depends on 1)

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.

### Claim 10 (depends on 1)

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.

### Claim 11

A method for reducing inflammation comprising administering a fibroin-derived protein composition to inflamed tissue;
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 value 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;
a plurality of peptide chains in the protein composition terminate in amide (—C(═O)NH2) groups;
the composition has a serine content that is reduced by greater than 25% compared to native fibroin protein, and wherein the serine content is at least about 5%; and
wherein the average molecular weight of the fibroin-derived protein composition is less than 60 kDa and greater than 10 kDa;
thereby reducing transcription factor signaling within cell nuclei of the tissue, thereby reducing the inflammation.

### Claim 12 (depends on 11)

The method of claim 11 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.

### Claim 13 (depends on 11)

The method of claim 11 wherein the administration is to the cornea and the administration reduces the presence of MMP-9 in the cornea.

### Claim 14 (depends on 11)

The method of claim 11 wherein the administration is to the eye and the administration reduces inflammation on the ocular surface.

### Claim 15 (depends on 11)

The method of claim 11 wherein the reduction in inflammation is accompanied by increased cell migration rates at the point of inflammation.

### Claim 16 (depends on 11)

The method of claim 11 wherein the protein composition has an average molecular weight less than 40 kDa.

### Claim 17 (depends on 11)

The method of claim 11 wherein the protein composition has an average molecular weight less than 35 kDa.

### Claim 18 (depends on 11)

The method of claim 11 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.

### Claim 19 (depends on 11)

The method of claim 11 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.

### Claim 20 (depends on 19)

The method of claim 19 wherein the inflammation is caused by dry eye syndrome.

### Claim 21

(canceled)

### Claim 22

(canceled)

### Claim 23

(canceled)