CN107405272B

CN107405272B - Cosmetic composition for skin regeneration comprising sinapic acid as index component of cynanchum atratum extract or cynanchum atratum extract having the same, and pharmaceutical composition for wound treatment

Info

Publication number: CN107405272B
Application number: CN201680013060.4A
Authority: CN
Inventors: 崔信旭; 洪仁基; 石地琄; 金诘; 裴炯珍; 金恩智
Original assignee: Radiant Inc
Current assignee: Radiant Inc
Priority date: 2015-04-13
Filing date: 2016-04-12
Publication date: 2023-06-09
Anticipated expiration: 2036-04-12
Also published as: CN107405272A; KR20160121847A; KR101768870B1; WO2016167527A1

Abstract

The present invention relates to a cosmetic composition for skin regeneration and a pharmaceutical composition for wound treatment, which contain sinapic acid or a cynanchum atratum extract having the same, and which control the occurrence of inflammation of the skin and exhibit excellent cell regeneration and wound treatment effects.

Description

Cosmetic composition for skin regeneration comprising sinapic acid as index component of cynanchum atratum extract or cynanchum atratum extract having the same, and pharmaceutical composition for wound treatment

Technical Field

The present invention relates to a cosmetic composition for skin regeneration and a pharmaceutical composition for wound treatment, which contain sinapic acid (sinapic acid) or a cynanchum atratum extract having sinapic acid (sinapic acid).

Background

The skin is an organ covering the outside of the human body, and has various functions of thermoregulation and barrier to the external environment. The skin is divided into three layers, namely a subcutaneous fat layer, dermis and epidermis from the innermost side. Wherein the subcutaneous fat layer is formed by fat cells and is located outermost in the skin layer. The dermis, which is formed of matrix proteins such as collagen fibers and elastic fibers, including blood vessels, nerves, sweat glands, etc., is located on the upper portion of the subcutaneous fat layer. The outermost epidermis is largely occupied by keratinocytes of the middle squamous epithelium.

However, such skin surfaces are continually damaged by deleterious environments such as ultraviolet light, dry weather, cold air, the outside of incorrect washing habits. Therefore, the outer skin cells constituting the outermost portion of the skin are continuously separated, and other cells below the outer skin cells continue to replace the outer skin cells.

As the relationship with the skin of modern people has increased, recently, in order to consider that such skin damage is induced, a peeling operation of removing a part of the skin portion to a large extent by using a chemical or laser has been attracting attention. The peeling operation is an operation for inducing the generation of new epidermis and stimulating the generation of collagen fibers of dermis, and is used for the treatment of acne, scars, wrinkles, pigmentation lesions, and treating wider pores. Such a peeling operation is a case where a wound is artificially generated in the skin, and the wound is all the states in which the continuity of the tissue is broken by external pressure.

When a wound occurs in the skin, the dermis is regenerated first and the epidermis is regenerated. More specifically, if the skin is wounded, the dermal fat cells decompose histamine to dilate the blood vessel, and inflammatory cells (leukocytes, platelets, etc.) are collected in the dilated blood vessel. If leukocytes in the aggregated inflammatory cells decompose cytokines, the fibrous subcellular components that maintain the elastic force of the dermis are activated, and collagen, mucopolysaccharide and other substances are secreted. Thereafter, the epidermis cleaved by the wound activates genes related to cell movement, and as a result, the epidermis regenerates.

One of the representative substances regulating such a wound treatment process may be growth factors (growth factors) which regulate the growth, differentiation, metabolism, etc. of cells and the surrounding environment of the wound through the whole process of the wound treatment. Among growth factors, epidermal growth factor (EGF, epidermal growth factor) is an important growth factor for in-bed therapy.

Epidermal growth factor has been studied to promote wound treatment of skin, cornea, end of year, etc. Also, the epidermal growth factor prevents the occurrence of scars, which are products occurring among adults, through wound treatment. For example, it has transforming growth factor beta (TGF-beta, transforming growth factor-beta). The transforming growth factor β is a substance involved in the growth and differentiation of various cells, and performs a complex function such as a plurality of complex functions (Bennett, n.t.et al., am.j. Surg.165:728,1993) including growth regulation, regulation of immune response, stimulation of bone formation, induction of cartilage-specific macromolecules (cartilage-specific macromolecule), and promotion of wound healing.

On the other hand, after laser treatment or peeling surgery, no inflammatory reaction is induced to regenerate skin using the above wound-treating substance. However, most of cosmetics developed so far for skin regeneration and wound treatment contain artificially prepared compound components in large amounts, and have a limited ability to stimulate skin side effects or skin regeneration from the beginning. Thus, recently, studies have been conducted to develop a natural product which has no side effects and is inexpensive to treat wounds.

However, cynanchum atratum extracts or sinapic acid have an effect on cell growth, differentiation, metabolism or trauma.

Disclosure of Invention

Technical problem

The present invention relates to a plant-derived substance, and particularly to a cosmetic composition for regenerating skin and a pharmaceutical composition for treating wounds, which are developed by developing a novel component for regenerating skin and treating wounds.

Means for solving the problems

In order to achieve the above object, the present invention provides a cosmetic composition for skin regeneration, which comprises sinapic acid (sinapic acid) as an active ingredient.

The present invention also provides a cosmetic composition for skin regeneration, which is characterized by comprising a plant extract containing sinapic acid as an active ingredient. In this case, the plant extract is preferably a cynanchum atratum (Cynanchum atratum) extract. The cynanchum atratum extract is obtained from cynanchum atratum by using one solvent selected from ethanol, dichloromethane and ethyl acetate, for example.

On the other hand, the cosmetic composition for skin regeneration of the present invention may contain ingredients commonly used in cosmetic compositions, such as antioxidants, stabilizers, solubilizers, vitamins, pigments and perfumes, as well as usual adjuvants and carriers, in addition to sinapic acid or plant extracts containing sinapic acid.

In the cosmetic composition for skin regeneration of the present invention, on the other hand, the cosmetic composition may include, as an example, a formulation selected from the group consisting of a solution, a suspension, an emulsion, a paste, a gel, a cosmetic composition, a lotion, a powder, a soap, a surfactant-containing cleanser, an oil, a foundation, an emulsified foundation, a wax base, and a spray.

In the case where the cosmetic composition of the present invention is in the form of a paste or gel, as the carrier component, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide or the like can be used.

In addition, in the case where the cosmetic composition of the present invention is in the form of a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and in the case of a spray, a propellant such as chlorofluorocarbon, propane/butane or dimethyl ether may be additionally contained.

In the case where the cosmetic composition of the present invention is in the form of a solution or emulsion, as the carrier component, a solvent, water as a solubilizer or an emulsifier, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1, 3-butanediol oil, glycerin fatty ester, polyethylene glycol or fatty acid ester of sorbitan may be used.

Also, in the case where the formulation of the cosmetic composition of the present invention is a suspension, as the carrier component, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth may be used.

In addition, in the case where the formulation of the cosmetic composition of the present invention is a surfactant-containing cleanser, as the carrier component, there can be used aliphatic alcohol sulfate, fatty alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazoline derivative, methyltaurine, sarcosinate, fatty acid amide ether sulfate, alkylamide betaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative or ethoxylated glycerol fatty acid ester.

In another aspect, the present invention provides a pharmaceutical composition for treatment, characterized by comprising sinapic acid as an active ingredient.

The present invention also provides a pharmaceutical composition for treatment, which is characterized by comprising a plant extract containing sinapic acid as an active ingredient. In this case, the plant extract may be preferably a cynanchum atratum extract. The cynanchum atratum extract is obtained from cynanchum atratum by using one selected from ethanol, dichloromethane and ethyl acetate as a solvent, for example.

On the other hand, the pharmaceutical composition for wound treatment of Benzhi may further comprise carriers, excipients and diluents commonly used in the preparation of pharmaceutical compositions. The pharmaceutical compositions for wound treatment of the present invention may be prepared in the form of powder, granule, tablet, capsule, suspension, emulsion, syrup, aerosol, etc. and sterile injection solution, or may be prepared in the form of cream, gel, patch, spray, ointment, coagulant, lotion, liniment, ointment or mucosal form, respectively, according to a general method, and may be used without limitation.

In the pharmaceutical composition for wound treatment of the present invention, as an example, lactose, glucose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil may be used as the carrier, excipient and diluent, and one or more selected from the group may be used.

In the pharmaceutical composition for wound treatment of the present invention, the solid preparation for oral administration may be, for example, granules, capsules, powders, granules, capsules, and may contain one or more selected from starch, calcium carbonate, sucrose or lactose, gelatin-form starch, calcium carbonate (calcium carbonate), sucrose (sucrose) or lactose (lactose), gelatin.

In the pharmaceutical composition for wound treatment of the present invention, the liquid phase preparation for oral administration may be a suspension, a liquid, an emulsion or a syrup, and may contain one or more selected from water, liquid paraffin, a wetting agent, a sweetener, a flavor and a preservative, as an example.

In the pharmaceutical composition for wound treatment of the present invention, the preparation for parenteral administration may be a sterilized aqueous solution, nonaqueous solvent, suspension, emulsion, or freeze-dried preparation, and may contain one or more selected from nonaqueous solutions, suspending agents such as propylene glycol (propylene glycol), polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.

On the other hand, the amount of the pharmaceutical composition for wound treatment of the present invention to be administered is determined in consideration of the administration method, the age, sex, weight, and extent of disease of the user, and the like. As an example, the pharmaceutical composition for wound treatment of the present invention is administered once a day in an amount of 0.0000001 to 100 mg/kg (body weight) based on the active ingredient. The above-described administration amount is merely an example, and may be changed according to the state of the user and the prescription of the doctor.

ADVANTAGEOUS EFFECTS OF INVENTION

The cosmetic composition for skin regeneration comprising sinapic acid (sinapic acid) or a cynanchum atratum extract having the same of the present invention exhibits excellent cell regeneration effect and wound treatment effect as well as a pharmaceutical composition for wound treatment.

Drawings

FIG. 1 is a graph of cell viability and growth capacity measured based on solvent species and concentration of Cynanchum atratum extract.

FIG. 2 is a graph of the ability to determine cell migration based on the solvent type and concentration of Cynanchum atratum extract.

FIG. 3 is a graph of the expression inhibition energy of "TNF- α" determined based on the solvent type and concentration of Cynanchum atratum extract.

FIG. 4 is a graph of "PGE" determined based on solvent type and concentration of Cynanchum atratum extract ₂ "graph of expression inhibition energy".

FIG. 5 is a graph showing the "IL-2" expression inhibition ability measured based on the concentration of the solvent species of Cynanchum atratum extract.

FIG. 6 is a graph showing the expression inhibition ability of "iNOS" and "COX-2" based on the solvent type and concentration of Cynanchum atratum extract.

FIG. 7 is a graph of cell viability and growth ability measured based on concentration of sinapic acid.

FIG. 8 is a graph of the measurement of cell growth capacity based on concentration of sinapic acid and time lapse.

FIG. 9 is a graph of the ability of cells to move based on concentration of sinapic acid.

FIG. 10 is a graph of the inhibition of expression of "TNF- α" as measured by concentration of sinapic acid.

FIG. 11 is a graph showing the inhibitory activity of "IL-2" expression based on concentration of sinapic acid.

FIG. 12 is a graph of "PGE" for determining mustard-based concentration ₂ "graph of expression inhibition energy".

Fig. 13 is a graph of the area of a wound site measured based on the addition of cynanchum atratum extract.

Detailed Description

The following examples, experimental examples and dosage form examples will be used to further describe the present invention in detail, but the scope of the present invention is not limited to the following examples and dosage form examples, but includes modifications of the technical ideas equivalent thereto.

Preparation example 1 preparation of Cynanchum atratum extract

The cynanchum atratum extract is prepared in this preparation example.

The dried whole herb of cynanchum atratum was purchased and prepared into powder by using a pulverizer. The powdered cynanchum atratum is extracted with 70% ethanol, 99.5% methylene chloride (dichlormethane) and 99% ethyl acetate (ethyl acetate) as extraction solvents, respectively. In this case, the amount of each extraction solvent used is 5 to 20ml per 1g dry weight of the cynanchum atratum powder. For extraction, extraction was performed at room temperature for 3 hours. After extraction, the extract was recovered with filter paper. The collected extracts were concentrated by a rotary vacuum concentrator, and then used in the following test.

( Experimental example 1: cytotoxicity and cell growth assay of cynanchum atratum extracts )

In this experimental example, "MTT assay" was performed on the cynanchum atratum extract prepared in preparation example 1 above to confirm cytotoxicity.

Human-derived keratinocytes (HaCaT) were seeded at a concentration of 1X 105cells/ml in 96-well plates and cultured. Thereafter, each of the cynanchum atratum extracts prepared in preparation example 1 was diluted to the concentration shown in table 1 below, and further cultured for 24, 48, and 72 hours by adding to the well plate. In this case, the survival rate and growth rate of cells were evaluated using an MTT reagent, and negative control groups were used as control groups in which no sample was treated.

TABLE 1

As a result of the experiment, the extract showed no toxicity in 25. Mu.g/ml, 50. Mu.g/ml, 99.5% methylene chloride 1. Mu.g/ml, 5. Mu.g/ml, 99% ethyl acetate 1. Mu.g/ml, 5. Mu.g/ml of 70% ethanol cynanchum atratum extract. In addition, cell growth can be achieved at these concentrations (FIG. 1).

Experimental example 2 measurement of cell motility of Cynanchum atratum extract

In this experiment, the movement of keratinocytes, which is an important mechanism, was confirmed during the treatment.

After keratinocytes (HaCaT) were inoculated and cultured at a concentration of 1X 105cells/ml in a 24-well plate, cells were extracted using a 200. Mu.l pipette tip. Thereafter, 25. Mu.g/ml, 50. Mu.g/ml, 99.5% methylene chloride 5. Mu.g/ml, and 99% ethyl acetate 5. Mu.g/ml of 70% ethanol cynanchum atratum extract which did not exhibit cytotoxicity in the above experimental example 1 were added, respectively, to measure cell migration for 72 hours. At this time, the control group utilized the epidermal growth factor having high cell mobility.

As a result of the experiment, the Cynanchum extract had excellent cell migration force compared with the EGF as a control group, and the Cynanchum extract exhibited the most excellent cell migration force among 50. Mu.g/ml of 70% ethanol Cynanchum extract (FIG. 2).

( Experimental example 3: evaluation of TNF-alpha expression inhibition ability of Cynanchum atratum extract )

Human-derived keratinocytes (HaCaT) were inoculated at a concentration of 1X 105cells/ml in a 24-well plate and cultured for 24 hours at a rate of 12.5mJ/cm ² Is processed at the ultraviolet B wavelength of (c). Thereafter, 25. Mu.g/ml, 50. Mu.g/ml, 99.5% methylene chloride 5. Mu.g/ml, and 99% ethyl acetate 5. Mu.g/ml of the 70% ethanol cynanchum atratum extract prepared in preparation example 1 were added to the well plates, respectively, and cultured for 24 hours. After incubation, the amount of TNF- α secreted in the culture broth was determined by immunoassay. At this time, the control group used 1. Mu.M dexamethasone (dexamethasone) as a positive control group.

As a result of the experiment, the expression of TNF- α produced by uv B in the cynanchum atratum extract was excellently inhibited compared to dexamethasone (fig. 3).

( Experimental example 4: evaluation of PGE2 expression inhibition ability of Cynanchum atratum extract )

Human-derived keratinocytes (HaCaT) were inoculated at a concentration of 1X 105cells/ml in a 24-well plate and cultured for 24 hours at a rate of 12.5mJ/cm ² Is processed at the ultraviolet B wavelength of (c). Thereafter, 25. Mu.g/ml, 50. Mu.g/ml, 99.5% methylene chloride 5. Mu.g/ml, 99% ethyl acetate 5. Mu.g/ml of the 70% ethanol cynanchum atratum extract prepared in preparation example 1 was added to the well plate, respectivelyAfter 24 hours of incubation, the amount of PGE2 secreted in the culture broth was determined by immunoassay. At this time, the control group used 1. Mu.M dexamethasone as a positive control group.

As a result of the experiment, the expression of PGE2 generated by ultraviolet rays B in the cynanchum atratum extract was excellently suppressed compared to dexamethasone (fig. 4).

( Experimental example 5: IL-2 expression inhibition ability evaluation of cynanchum atratum extract )

Human-derived keratinocytes (HaCaT) were inoculated at a concentration of 1X 105cells/ml in a 24-well plate and cultured for 24 hours at a rate of 12.5mJ/cm ² Is processed at the ultraviolet B wavelength of (c). Thereafter, 25. Mu.g/ml, 50. Mu.g/ml, 99.5% methylene chloride 5. Mu.g/ml, and 99% ethyl acetate 5. Mu.g/ml of the 70% ethanol cynanchum atratum extract prepared in preparation example 1 were added to the well plate to culture the mixture for 24 hours, and the amount of IL-2 secreted in the culture broth was measured by immunoassay. At this time, the control group used 1. Mu.M dexamethasone as a positive control group.

As a result of the experiment, the expression of IL-2 produced by ultraviolet B in the cynanchum atratum extract was excellently inhibited compared to dexamethasone (fig. 5).

( Experimental example 6: evaluation of inhibition ability of iNOS and COX-2 expression of Cynanchum atratum extract )

Human-derived keratinocytes (HaCaT) were inoculated at a concentration of 1X 105cells/ml in a 60-well plate and cultured for 24 hours at a rate of 12.5mJ/cm ² Is processed at the ultraviolet B wavelength of (c). Thereafter, 25. Mu.g/ml, 50. Mu.g/ml, 99.5% methylene chloride 5. Mu.g/ml, and 99% ethyl acetate 5. Mu.g/ml of the 70% ethanol cynanchum atratum extract prepared in preparation example 1 were added to the well plates, respectively, and cultured for 24 hours. After the cultivation, proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE, sodiumdidecylsulfate-polyacrylamide gel electrophoresis), and after shifting to PVDF membrane (membrane) by "electrotransfer", intracellular expression levels were confirmed by immunoblotting (immunoblotting). At this time, the control group used 1. Mu.M dexamethasone as a positive control group.

As a result of the experiment, the expression of iNOS and COX-2 produced by ultraviolet ray B in the cynanchum atratum extract was excellently inhibited as compared with dexamethasone (FIG. 6).

( Experimental example 7: cytotoxicity and cell growth assay of sinapic acid )

In this experimental example, MTT assay was performed on sinapic acid in order to confirm cytotoxicity of sinapic acid as an index component of the cynanchum atratum extract prepared in the above experimental example 1. In this case, sinapic acid was confirmed to be an index component of the cynanchum atratum extract prepared in the above-mentioned preparation example 1, which was requested by the institute of new natural science and medicine, kyoto, science and technology.

Human-derived keratinocytes (HaCaT) were prepared at 1X 10 in 96-well plates ⁵ After inoculation and cultivation at the concentration of cells/ml, the concentration of sinapic acid was diluted to 25, 50, 100, 200, 250, 500. Mu.M, respectively, and added to the well plate for cultivation for 24 hours. After incubation, the viability of the cells was assessed using MTT reagents. In this case, a negative control group in which no sample was treated was used as the control group.

As a result of the experiment, sinapic acid did not exhibit cytotoxicity at the concentrations of 25, 50, 100, 200, 250, 500. Mu.M (FIG. 7).

( Experimental example 8: cell growth assay based on sinapic acid addition concentration and time lapse )

Human-derived keratinocytes (HaCaT) were prepared at 1X 10 in 96-well plates ⁵ After inoculation and cultivation at the concentration of cells/ml, the concentration of sinapic acid was diluted to 25, 50, 100, 200, 250, 500. Mu.M and added to the well plate for cultivation for 24, 48, 72 hours. After incubation, the growth rate of the cells was evaluated using MTT reagent. In this case, a negative control group, in which no sample was treated, was used as the control group, and an epidermal growth factor was used as the positive control group.

As a result of the experiment, the concentration of sinapic acid was relied upon to achieve cell growth. In the case of the concentration of 100. Mu.M or more, the cell growth was higher in the case of culturing for 24 hours than in the case of culturing for 48 hours and 72 hours (FIG. 8).

Experimental example 9 determination of the cell movement ability of sinapic acid

In this experimental example, the movement of keratinocytes, which is an important mechanism during wound treatment, was confirmed.

Keratinocytes (HaCaT) were plated at 1X 10 in 24-well plates ⁵ After inoculation and incubation at a concentration of cells/ml, the cells were harvested using a 200. Mu.l pipette tip. After that, sinapic acid was added to the well plate and cell movement was measured for 72 hours. The control group utilized epidermal growth factor that increased the ability of the cells to move.

As a result of the experiment, the cell mobility was increased by sinapic acid (fig. 9).

Experimental example 10 evaluation of the inhibitory Activity of sinapic acid against TNF-alpha expression

Human-derived keratinocytes (HaCaT) were prepared at 1X 10 in 24-well plates ⁵ After inoculation and incubation at a cell/ml concentration for 24 hours, the cells were inoculated at a rate of 12.5mJ/cm ² After treatment with ultraviolet B-wavelength, sinapic acid was added at 50.100. Mu.M and incubated for 24 hours, and the amount of TNF-. Alpha.secreted in the culture broth was measured by immunoassay. At this time, the control group used 1. Mu.M dexamethasone as a positive control group.

As a result of the experiment, sinapic acid inhibited TNF- α expression similarly to dexamethasone as a positive control group (fig. 10).

Experimental example 11 evaluation of expression inhibition ability of sinapic acid IL-2

Human-derived keratinocytes (HaCaT) were prepared at 1X 10 in 24-well plates ⁵ The cells/ml concentration was inoculated and cultured for 24 hours. According to 12.5mJ/cm ² After the ultraviolet B-wavelength treatment, sinapic acid was added at 50 and 100. Mu.M and incubated for 24 hours. After the culture, the amount of IL-2 secreted in the culture broth was determined by immunoassay. At this time, the control group used 1. Mu.M dexamethasone as a positive control group.

As a result of the experiment, sinapic acid inhibited IL-2 expression similarly to dexamethasone as a positive control group (fig. 11).

Experimental example 12 evaluation of sinapic acid PGE2 expression inhibition ability

Human-derived keratinocytes (HaCaT) were prepared at 1X 10 in 24-well plates ⁵ concentration of cells/mlInoculated and cultured for 24 hours. According to 12.5mJ/cm ² After the ultraviolet B-wavelength treatment, sinapic acid was added at 50 and 100. Mu.M and incubated for 24 hours. After incubation, PGE secreted in the broth was assayed by immunoassay ₂ Is a combination of the amounts of (a) and (b). At this time, the control group used 1. Mu.M dexamethasone as a positive control group.

As a result of the experiment, sinapic acid inhibited PGE similarly to dexamethasone as a positive control group ₂ Is shown (FIG. 12).

( Experimental example 13: evaluation of skin wound treatment efficacy of Cynanchum atratum extract )

In this experimental example, the cynanchum atratum extract extracted using 70% ethanol in the above preparation example 1 was used to evaluate the skin wound treatment efficacy. Hereinafter, in the following experimental examples, experiments were performed using the cynanchum atratum extract as used in the present experimental examples.

For experiments, 5 weeks male SD rate was used in the Zoak fish widely used for toxicity and drug efficacy experiments, 5 weeks male SD rate was anesthetized for preparation of a wearing model, and after dehairing of equivalent parts, skin was sterilized with 70% ethanol and povidone. Using the vertebra county as the midline, a 3cm mesh was used on one side to initiate the trauma and removed to the dermis. Thereafter, the wound site was directly coated with cynanchum atratum extracts at concentrations of 0.5% (w/v), 1% (w/v) and 2% (w/v), respectively, and then Dressing (Dressing) was stably performed.

After the wound site was coated once daily with the test substance, the wound site was photographed at 2-time intervals per week, and the wound site area was measured using ImageJ soft for 24 days. The results are presented in table 2 below.

TABLE 2

As a result of the experiment, the recovery rate and the area were different between the 0.5% (w/v), 0.1% (w/v), and 2% (w/v) cynanchum atratum extract-treated group and the comparative group from 10 days later, and were superior to those of the comparative group from 10 days later to 24 days later.

( Experimental example 14: evaluation of skin wound treatment histopathological efficacy of cynanchum atratum extract )

After performing a Hematoxylin & Eosin (H & E) staining in order to evaluate the histopathological efficacy of the Cynanchum atratum extract for skin wound treatment, observation was performed using an optical microscope.

To compare the treatment profiles of the individual experimental groups, the conditions of inflammation (infusion), fibroplasia (Fiboroplasia), epithelialization (epithelialization) were determined, starting from 0 to 3 points from which the corresponding conditions were not observed, and a total of 4 steps were divided for evaluation. The evaluation results are shown in table 3 below.

TABLE 3 Table 3

As shown in table 3, the inflammation was alleviated, and the combined tissue proliferation and epithelialization were increased by the cynanchum atratum extract treatment group.

( Experimental example 1: cream-type cosmetic composition for skin regeneration comprising cynanchum atratum extract as active ingredient )

In this example, a cream-type cosmetic composition for skin regeneration containing a cynanchum atratum extract as an active ingredient was prepared. The ingredients containing the cynanchum atratum extract with a solid content of 2% (w/v) were mixed in accordance with the contents of the following table 4 to prepare a cream-type cosmetic composition for skin regeneration.

TABLE 4 Table 4

Composition of the components	Content (weight percentage)
		2% (w/v) Cynanchum atratum extract	4.0
Glycerol	2.0
		Parabens	0.2
Allantoin	2.0
		Betaine (betaine)	3.0
Sodium hyaluronate	1.0
		Tocopheryl acetate	5.0
Butter resin	2.0
		Trehalose	1.0
Preservative and perfume	Proper amount of
		Purified water	Near 100
Totalizing	100

On the other hand, in the following examples, the same cynanchum atratum extract as in the present example was used.

( Experimental example 2: pharmaceutical composition for wound treatment comprising cynanchum atratum extract as active ingredient )

In this example, a pharmaceutical composition (ointment) for wound treatment comprising a cynanchum atratum extract as an active ingredient was prepared.

The ingredients of the present invention containing a cynanchum atratum extract with a solid content of 2% (w/v) were mixed at the contents of the following table 5 to prepare a pharmaceutical composition (ointment) for wound treatment.

TABLE 5

Composition of the components	Content (weight percentage)
		2% (w/v) Cynanchum atratum extract	10
Sebacic acid diethyl ester	8
		Whale wax	5
Polyoxyethylene oleyl ether phosphate	6
		Sodium benzoate	Proper amount of
VaselineForest stand	Near 100
		Totalizing	100

( Experimental example 3: cream-type cosmetic composition for skin regeneration containing sinapic acid as active ingredient )

In this example, a cream-type cosmetic composition for skin regeneration containing sinapic acid as an active ingredient was prepared. A cream-type cosmetic composition for skin regeneration was prepared by mixing ingredients containing 2% (w/v) sinapic acid in the amounts shown in Table 6 below.

TABLE 6

Composition of the components	Content (weight percentage)
		2% (w/v) Cynanchum atratum extract	0.4
Glycerol	2.0
		Parabens	0.2
Allantoin	2.0
		Betaine (betaine)	3.0
Sodium hyaluronate	1.0
		Tocopheryl acetate	5.0
Butter resin	2.0
		Trehalose	1.0
Preservative and perfume	Appropriately, is
		Purified water	Near 100
Totalizing	100

On the other hand, in the following examples, the same sinapic acid as in this example was used.

( Example 4: pharmaceutical composition for wound treatment comprising sinapic acid as active ingredient )

The ingredients of the present invention containing a cynanchum atratum extract with a solid content of 2% (w/v) were mixed at the contents of the following table 7 to prepare a pharmaceutical composition (ointment) for wound treatment.

TABLE 7

Composition of the components	Content (weight percentage)
		2% (w/v) Cynanchum atratum extract	1
Sebacic acid diethyl ester	8
		Whale wax	5
Polyoxyethylene oleyl ether phosphate	6
		Sodium benzoate	Proper amount of
Vaseline	Near			100
		Totalizing	100

Claims

1. Use of a cynanchum atratum (Cynanchum atratum) extract for the preparation of a cosmetic composition for the regeneration of wound skin, the cynanchum atratum extract comprising sinapic acid as an active ingredient and having cell migration ability, and the cynanchum atratum extract being an extract obtained from cynanchum atratum using 70% ethanol as a solvent, wherein the regeneration of wound skin comprises that inflammation of skin is alleviated, connective tissue hyperplasia and epithelialization is increased,

wherein, in the cosmetic composition, the cynanchum atratum extract is an active ingredient that promotes regeneration of wound skin, which is capable of alleviating inflammation of skin, promoting connective tissue proliferation, and increasing epithelialization.