# US 2018/0021385 A1 · Wound healing formulation

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- Number: US 2018/0021385 A1
- Document ID: US20180021385A1
- Status: Published application
- Kind: A1
- Application number: 15/548,666
- Filed: 2015-12-22
- Published: 2018-01-25
- Earliest claimed: 2015-02-04
- Applicants: Stichting VUmc; A-Skin Holding B.V.
- Inventors: Susan GIBBS; Riekeld Johannes SCHEPER
- Web page: https://patentgrep.com/patent/US20180021385A1

## CPC

- A61K 35/36: Skin; Hair; Nails; Sebaceous glands; Cerumen; Epidermis; Epithelial cells; Keratinocytes; Langerhans cells; Ectodermal cells (islets of Langerhans A61K35/39)
- A61K 35/33: Fibroblasts
- A61P 17/02: for treating wounds, ulcers, burns, scars, keloids, or the like
- C12N 5/0629: {Keratinocytes; Whole skin}
- C12N 5/0656: {Adult fibroblasts}
- C12N 2502/094: keratinocytes
- C12N 2502/1323: Adult fibroblasts
- 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
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present disclosure relates to a method for producing a composition, wherein the method comprises the steps of co-culturing immortalized fibroblasts and immortalized keratinocytes, thereby producing secretion; separating the secretion from the fibroblasts and keratinocytes; and providing a pharmaceutically acceptable composition comprising the secretion. The present disclosure also relates to the composition obtainable by the method, wherein the composition preferably is a pharmaceutical composition for medical use, preferably for use in the treatment of a wound, preferably a chronic or acute wound.

## Claims

### Claim 1

Method for producing a therapeutic wound healing composition, wherein the method comprises the steps of:
a) co-culturing immortalized fibroblasts and immortalized keratinocytes, thereby producing secretion;
b) separating the secretion from the fibroblasts and keratinocytes; and
c) providing a therapeutic wound healing composition comprising the secretion.

### Claim 2 (depends on 1)

Method according to claim 1, wherein in step a) the percentage of the immortalized fibroblasts relative to the total number of immortalized fibroblasts and immortalized keratinocytes is between 5% and 95%.

### Claim 3 (depends on 1)

Method according to of claims 1, wherein
the immortalized fibroblasts are TERT-immortalized fibroblasts; and/or
the immortalized keratinocytes are A431 keratinocytes or TERT-immortalized keratinocytes.

### Claim 4 (depends on 1)

Method according to claims 1, wherein step a) is performed
in vitro;
at 30-40° C.;
for at least 1 hour; and/or
in a clinical grade medium.

### Claim 5 (depends on 1)

Method according to claim 1, wherein in step a)
the immortalized fibroblasts and immortalized keratinocytes are arranged randomly in the co-culture; and/or
the immortalized fibroblasts and immortalized keratinocytes are co-cultured together in a monolayer of cells.

### Claim 6 (depends on 1)

A composition obtainable by the method according to claim 1, wherein the composition is a pharmaceutically acceptable composition.

### Claim 7 (depends on 6)

A method for the treatment of a wound, comprising applying the composition of claim 6 to the wound in an amount effective for the treatment of the wound.

### Claim 8 (depends on 7)

The method according to claim 7, wherein the treatment of a wound comprises applying the composition onto a skin wound.

### Claim 9 (depends on 6)

The composition according to claim 6, wherein the composition contains no fibroblasts and/or keratinocytes.

### Claim 10 (depends on 6)

The composition according to claim 6, wherein the composition is in the form of a liquid, ointment, cream, gel, hydrogel, lotion, dressing, or carrier.

### Claim 11 (depends on 1)

Method according to claim 1, wherein co-culturing immortalized fibroblasts and immortalized keratinocytes, thereby producing secretion, provides a percentage of the immortalized fibroblasts relative to the total number of immortalized fibroblasts and immortalized kerotinocytes between 25% and 95%.

### Claim 12 (depends on 2)

Method according to claim 2, wherein
the immortalized fibroblasts are TERT-immortalized fibroblasts; and/or
the immortalized keratinocytes are A431 keratinocytes or TERT-immortalized keratinocytes.

### Claim 13 (depends on 4)

Method according to claim 4, wherein co-culturing immortalized fibroblasts and immortalized keratinocytes, thereby producing secretion, is performed at 34-38° C.

### Claim 14 (depends on 4)

Method according to claim 4, wherein co-culturing immortalized fibroblasts and immortalized keratinocytes, thereby producing secretion, is performed for at most 240 hours.

### Claim 15 (depends on 1)

Method according to claim 1, wherein co-culturing immortalized fibroblasts and immortalized keratinocytes, thereby producing secretion, is performed
in vitro;
at 34-38° C.;
for 1 hour to 240 hours; and
in a clinical grade medium.

### Claim 16 (depends on 8)

The method of claim 8, wherein the applying the composition onto a skin wound is followed by applying a dressing, compression therapy, or a tissue substitute or an autograft onto the skin wound.