KR102503612B1 - Skin external composition - Google Patents

Abstract

The present invention relates to a composition for external application for skin including an extract of Scrophularia takesimensis Nakai callus as an active ingredient. The present invention includes an extract of Scrophularia takesimensis Nakai callus extracted by a hot water extraction method as an active ingredient. The present invention provides a skin anti-inflammatory effect, a skin barrier improvement effect and a skin moisturizing effect while being safe to a human body with few side effects.

Description

Composition for external application for skin {SKIN EXTERNAL COMPOSITION}

It relates to a composition for external application for skin.

The skin is composed of epidermis, dermis, and subcutaneous fat layer, and the keratinocytes of the epidermis and lipids between keratinocytes act as a barrier to prevent body fluid loss and protect the human body from external harmful factors. do

The skin barrier can maintain its function only when the stratum corneum located at the top of the epidermis is normally formed. The stratum corneum, located at the outermost part of the skin and in contact with the outside, plays an important role in the skin barrier function even though the cells are in a dead state. In other words, the stratum corneum provides an environment in which the skin can maintain normal biological functions by suppressing the loss of water and electrolytes through the skin, and protects against external harmful factors such as toxic substances, microorganisms, mechanical stimulation, and ultraviolet rays that invade through the skin. It serves as the first line of defense.

The stratum corneum of the skin is composed of about 40% protein, 40% water, and 10% to 20% lipids. The structure is composed of protein-rich corneocytes and lipids filling the gap, among which lipids play a major role as barriers.

Keratinocytes are mainly composed of keratin bundles and are involved in the structural stability and elasticity of the skin. Unlike other common biomembranes, most lipids are composed of ceramide, cholesterol, and free fatty acids rather than phospholipids. Ordinary biomembranes containing phospholipid, sphingomyelin, and cholesterol do not perform well as barriers because they are easily soluble in water or water and highly permeable to substances with low molecular weight. However, in the stratum corneum, ceramide Since lipids such as lipids, cholesterol, and free fatty acids are well connected in a straight line, they act as a barrier to prevent permeation of water and hydrophilic substances.

When the skin barrier is damaged, it is easily exposed to various allergens penetrating from the outside. The skin with the damaged skin barrier increases the skin pH, increases the activity of serine protease, and decomposes the protein of keratinocytes. Enzyme activated receptor-2 (PAR-2) is activated to secrete IL-25, IL-33, and thymic stromal lymphopoietin (TSLP) out of the cell. These factors activate Th2 immune cells and exacerbate inflammatory reactions such as atopic dermatitis.

On the other hand, the function of the skin barrier is greatly influenced by the composition and organization of lipids. Lipids between keratinocytes are composed of ceramide, cholesterol, and free fatty acids at the same molar ratio. It has been reported that ceramide is reduced in skin with damaged skin barriers such as dry skin, atopic dermatitis, acne skin, and aging skin. there is. In the skin of patients with atopic dermatitis, abnormal skin barrier function is observed, transepidermal water loss (TEWL) is increased by 2 to 5 times, and the water content of the stratum corneum is reduced, which is due to a decrease in stratum corneum lipids, In particular, the decrease in ceramide has been found to be the main cause.

In order to improve the skin barrier in such skin, various types of topical treatments using ceramides (or synthetic pseudo-ceramides) have been attempted. However, natural ceramides are expensive and have limitations in use due to safety controversy, and synthetic ceramides identical or similar to natural ceramides also have problems in that it is difficult to introduce them into various formulations due to high prices and low solubility.

One embodiment provides a composition for external application for skin that is safe to the human body with few side effects and has excellent skin anti-inflammatory effect, skin barrier improvement, and skin moisturizing effect.

One embodiment provides a composition for external application for skin comprising a callus extract of ginseng ginseng as an active ingredient.

The ginseng callus extract may be obtained by extracting the ginseng callus, and the ginseng callus may be obtained by culturing and inducing leaves, stems, roots, flowers, or a combination thereof in an induction medium.

The concentration of the ginseng callus extract may be 0.1 μg/mL to 500 μg/mL.

The ginseng callus extract may be included in an amount of 0.0001% to 10% by weight based on the total amount of the composition for external application for skin.

The ginseng callus extract can inhibit the expression of inflammatory cytokines.

The ginseng callus extract can promote ceramide biosynthesis.

The ginseng callus extract can activate serine palmitoyl transferase (SPT), ceramide synthase (CerS), or a combination thereof.

The composition for external application for skin may be for improving skin barrier, for moisturizing skin, for relieving skin inflammation, or a combination thereof.

The composition for external application for skin may be a cosmetic composition, a hair composition for improving the scalp, a pharmaceutical composition for improving the skin barrier and moisturizing the skin, or a combination thereof.

The composition for external application for skin according to an embodiment has few side effects, is safe to the human body, and has excellent skin anti-inflammatory effects, skin barrier improvement and skin moisturizing effects, and is thus used as a cosmetic composition, a hair composition for improving the scalp, and a pharmaceutical composition for improving the skin barrier and moisturizing the skin. can be useful

1 is a graph showing the degree of inhibition of the expression of the inflammatory cytokine IL-25 when keratinocytes were treated with an extract of ginseng ginseng callus according to one embodiment.
2 is a graph showing the degree of increase in ceramide biosynthesis when keratinocytes were treated with an extract of ginseng callus according to one embodiment.
3 is a graph showing the degree of change in transepidermal water loss when the compositions for external application for skin according to Example 1 and Comparative Example 1 were applied.

Hereinafter, embodiments will be described in detail so that those skilled in the art can easily implement them. However, implementations may take many different forms and are not limited to the implementations described herein.

The composition for external application for skin according to one embodiment may include an extract of ginseng callus as an active ingredient.

When the ginseng callus extract is used in a composition for external application for skin, an excellent skin anti-inflammatory effect can be obtained by suppressing the expression of inflammatory cytokines, and also promotes ceramide biosynthesis and increases the gene expression of enzymes related to ceramide production to improve skin barrier function It can improve and obtain excellent skin moisturizing effect.

The ginseng callus extract may be obtained by extracting ginseng callus.

Scrophularia takesimensis Nakai, which is used to obtain the callus of ginseng ginseng, is a perennial plant belonging to the genus ginseng. It is endemic to Korea and is distributed only on Ulleungdo Island. It lives in crevices of coastal gravel or stone embankments along roads, but young individuals also live in sandy areas.

The ginseng callus can be obtained by culturing and inducing the leaves, stems, roots, flowers, or a combination thereof of ginseng ginseng in an induction medium, and may include a culture solution in which a part of ginseng ginseng is cultured.

The induction of the callus of ginseng using the ginseng can be carried out through a conventional method known in the art.

According to one embodiment, it is possible to obtain a raw material of an effective substance without damaging the plant by using the ginseng callus or the cells differentiated from the ginseng callus, rather than using the plant itself, which is superior to the plant itself. can hold

Methods for extracting the callus of ginseng ginseng include hot water extraction, cold extraction, solvent extraction, steam distillation, elution, compression, ultrasonic extraction, filtration, reflux extraction, etc., which are performed alone or in combination with two or more methods can do.

Among the above methods, for example, hot water extraction or solvent extraction may be used to extract the callus of ginseng. The solvent extraction method is extraction using water, an organic solvent, or a mixture thereof, and an organic solvent may be used, for example.

The organic solvent may be C1 to C4 alcohol, and may be, for example, at least one selected from the group consisting of methanol, ethanol, propyl alcohol, butyl alcohol, glycerin, propylene glycol, and butylene glycol.

The ginseng callus extract can be obtained by a solvent extraction method in which dried ginseng callus is pulverized, an extraction solvent is added, and the resulting solution is filtered and recovered after stirring at appropriate intervals while depositing for a predetermined time.

The amount of the extraction solvent may be 1 to 50 times the total volume of the ginseng callus, for example, 5 ml to 20 ml per 1 g of dry weight of the ginseng callus. Immersion extraction may be performed at room temperature for 1 to 7 days, or extraction may be performed for 1 hour to 5 hours at a temperature of 50° C. to 95° C. When extracted in the above temperature range, it is effective in preventing decay, discoloration, and odor, and when extracted in the above time range, extraction is performed well to secure sufficient active ingredients.

The concentration of the ginseng callus extract may be 0.1 μg/mL to 500 μg/mL, for example, 0.4 μg/mL to 400 μg/mL, and 1 μg/mL to 400 μg/mL. . When the concentration of ginseng callus extract is within the above range, the expression of inflammatory cytokines can be suppressed, improving the skin anti-inflammatory effect, promoting ceramide biosynthesis and increasing the gene expression of enzymes related to ceramide production, thereby strengthening the skin barrier. and skin moisturizing effect can be improved.

The ginseng callus extract may be included in 0.0001% to 10% by weight based on the total amount of the composition for external application for skin, for example, 0.001% to 5% by weight, 0.01% to 5% by weight, 0.1% to 0.1% by weight 1% by weight. When the ginseng callus extract is included within the above content range, the stability of the formulation is improved, the fragrance is excellent, and the skin anti-inflammatory effect, skin barrier improvement, and skin moisturizing effect are excellent. Cosmetic composition, hair composition for improving scalp, skin barrier improvement And it can be usefully used as a pharmaceutical composition for skin moisturizing.

The ginseng callus extract according to one embodiment can suppress the expression of inflammatory cytokines and promote ceramide biosynthesis. In addition, enzymes related to ceramide production may be activated, for example, serine palmitoyl transferase (SPT), ceramide synthase (CerS), or a combination thereof may be activated.

Accordingly, the composition for external application for skin according to one embodiment may be used for improving skin barrier, moisturizing skin, relieving skin inflammation, or a combination thereof.

The composition for external application for skin according to one embodiment may further include additives in addition to the ginseng callus extract, depending on the use. The additives may be used alone or in combination with a moisturizer, a thickener, a surfactant, an oil base, a preservative, an antioxidant, an alcohol, a flavoring agent, a pH adjuster, or a natural extract.

The composition for external application for skin according to one embodiment may be a cosmetic composition, a hair composition for improving the scalp, a pharmaceutical composition for improving the skin barrier and moisturizing the skin, or a combination thereof.

The formulation of the cosmetic composition is not particularly limited and may be appropriately selected according to the purpose. For example, it may be prepared in formulations such as softening lotion, astringent lotion, nutrient lotion, massage cream, essence, nutrient cream, pack, powder, and waxy cream, but is not limited thereto.

The hair composition for improving the scalp may be in formulations such as shampoo, rinse, treatment, and hair pack, but is not limited thereto.

The pharmaceutical composition may be a transdermal dosage form such as lotion, ointment, gel, cream, patch, etc., but is not limited thereto.

The pharmaceutical composition can be used for skin barrier-related diseases, for example, atopic dermatitis, dermatitis due to rough skin, heat rash, soreness, frostbite, diaper dermatitis, contact dermatitis, seborrheic dermatitis, acne, Lichen Vidal, nummular eczema, housewives' eczema, solar dermatitis, ichthyosis, prurigo dermatitis, prurigo, drug rash, toxic rash, psoriasis, psoriasis psoriasis, palmar pustulosis, lichen planus, lichen glaucous pilaris erythematosus, Gibert's rosea rain Dermatosis, erythematosis, erythematosis, discoid lupus erythematosus, systemic lupus erythematosus, pemphigus, pemphigoid, during herpetic dermatitis, alopecia areata, vitiligo vulgaris, sarcoidosis, cutaneous amiloridesis, keloids, hypertrophic hemiplegia, It can be used for diseases such as wounds, bedsores, skin ulcers, hair loss, wool, and hair growth.

Hereinafter, specific embodiments of the present invention are presented. However, the embodiments described below are only intended to specifically illustrate or explain the present invention, and the present invention should not be limited thereto. In addition, since the content not described herein can be sufficiently technically inferred by those skilled in the art, the description thereof will be omitted.

Preparation Example 1: Preparation of ginseng callus extract

After sterilizing the leaves of Seomyeonsam (Obtainer: Ulleung-eup, Ulleung-gun, Gyeongsangbuk-do), the leaves were cut with a knife and cultured in MS medium to induce callus of Seomyeonsam. After harvesting and drying the ginseng callus, 4 g of the dried ginseng callus powder was placed in a container, 100 mL of purified water was added, and hot water extraction was performed at 121 ° C for 15 minutes. After extraction, the solids were removed by filtration through a mesh to prepare a callus extract of ginseng ginseng.

Experimental Example 1: Assessment of inhibition of inflammatory cytokine (IL-25) expression

In order to evaluate the inhibition of inflammatory cytokine expression by the ginseng callus extract according to Preparation Example 1, human keratinocytes were treated with 10% FBS (fetal bovine serum) and DMEM (Dulbecco Modified Eagle Medium) medium and inoculated into a 6-well plate at a concentration of 5×10 ⁴ cells/well, and after 24 hours, PAR-2 agonist 10 μM and ginseng callus extract at 0.4 μg/mL each, Concentrations of 4 μg/mL, 40 μg/mL and 400 μg/mL were applied to human keratinocytes. After 48 hours of culture, the supernatant was taken and the IL-25 expression level was measured using the Interleukin-25 ELISA kit (Abcam, USA). At this time, as a negative control group, a group not treated with either PAR-2 agonist or ginseng callus extract was used.

The results are shown in Figure 1. 1 is a graph showing the degree of inhibition of the expression of the inflammatory cytokine IL-25 when keratinocytes were treated with an extract of ginseng ginseng callus according to one embodiment. Referring to Figure 1, it was confirmed that the expression of the inflammatory cytokine IL-25 was inhibited by the callus extract of ginseng ginseng according to one embodiment.

Experimental Example 2: Evaluation of enzyme expression related to ceramide production process

In order to evaluate the expression of enzymes related to ceramide production by the ginseng callus extract according to Preparation Example 1, first, human keratinocytes were cultured in a DMEM medium containing 10% FBS and 1% P/S to form 2.5 × 10 After inoculation in a 6-well plate at a concentration of ⁵ cells/well, 24 hours later, the callus extract of 0.4 μg/mL, 4 μg/mL, 40 μg/mL, and 400 μg/mL respectively was applied to human keratinocytes.

To measure gene expression levels, total RNA was extracted from cells using easy-spin™ Total RNA extraction kit (iNtRoN, Korea). For the extracted RNA, cDNA was synthesized using Maxime RT PreMix (iNtRON, Korea), and real-time PCR was performed using ABI 7500 Fast Real-Time PCR system (Applied Biosystems, USA) to generate serine palmitoyl transferase (serine palmitoyl transferase). transferase (SPT) and ceramide synthase (CerS) gene expression levels were confirmed. At this time, as a negative control group, a group not treated with the callus extract of ginseng ginseng was used, and GAPDH mRNA was used as an internal control group.

The gene expression level was expressed as a ratio compared to the value of the control group (control-treated = 1), and the results are shown in Table 1 below.

(control-treated = 1) SPT1 SPT2 CerS3 ginseng callus extract 0.4 μg/mL 1.2 1.4 1.3 ginseng callus extract 4 μg/mL 1.4 1.5 1.4 ginseng callus extract 40 μg/mL 1.5 1.8 1.8 ginseng callus extract 400 μg/mL 1.6 2.0 1.9

Through Table 1, it was confirmed that the gene expression of enzymes SPT and CerS related to ceramide production increased by the treatment of the callus extract of ginseng ginseng according to one embodiment.

Experimental Example 3: Test for confirming increase in ceramide production

In order to measure the effect of increasing the amount of ceramide produced by the ginseng callus extract, human keratinocytes were cultured in a DMEM medium containing 10% FBS and 1% P/S to obtain 2.5×10 ⁵ cells/well (cells/well). Well), human keratinocytes were inoculated into a 6-well plate at a concentration of 0.4 µg/mL, 4 µg/mL, 40 µg/mL, and 400 µg/mL, respectively, after 24 hours. applied to. After culturing for 48 hours, the amount of ceramide eluted in the medium was measured using a Ceramide ELISA assay kit (Mybiosource, USA). As a negative control group, a group not treated with ginseng callus extract was used.

The results are shown in FIG. 2 . 2 is a graph showing the degree of increase in ceramide biosynthesis when keratinocytes were treated with an extract of ginseng callus according to one embodiment. Referring to Figure 2, it was confirmed that the amount of ceramide production increased by the ginseng callus extract according to one embodiment.

Experimental Example 4: Safety test on skin

In order to evaluate the degree of irritation to the skin, a human patch test was performed. The skin patch test was conducted on 30 healthy people over the age of 20, and samples were made with 0.1% by weight, 1% by weight and 10% by weight aqueous solutions, and then applied to the inner part of the upper arm for 48 hours using a Finn chamber. The first reading was taken after removal after attachment, and the second reading was performed after 72 hours had elapsed to determine the skin reaction result. The evaluation criteria and the calculation of the stimulation index are shown in Table 2 and Equation 1 below, and the results are shown in Table 3 below.

reaction weight Evaluation standard - 0.0 unresponsive ± 0.5 suspected positive reaction + 1.0 weak positive reaction ++ 2.0 strong positive reaction +++ 3.0 polar positive reaction

[Equation 1]

Skin irritation index = Σ (weight X number of responders) / (highest weight X total number of subjects) X 100

skin irritation index degree of irritation ginseng callus extract 0.1% by weight 0.0 non-irritating ginseng callus extract 1% by weight 0.0 non-irritating ginseng callus extract 10% by weight 0.0 non-irritating

Referring to Table 3, it was found that the ginseng callus extract was non-irritating to the skin at 10% by weight or less, and it was determined that there was no irritation to the skin when used as a raw material for external skin preparations.

Experimental Example 5: Skin barrier improvement test through transepidermal water loss (TEWL) measurement

In order to confirm the skin barrier improving effect of the composition for external application for skin according to one embodiment, the composition for external application for skin, specifically the cream composition, of Example 1 and Comparative Example 1 was prepared as shown in Table 4 below.

ingredient name Content (% by weight) Example 1 Comparative Example 1 Ginseng Callus Extract 0.5 - glycerin 5.0 5.0 Stearic Acid 8.0 8.0 squalane 5.0 5.0 Self-Emulsifying Glycerin Monostearate 2.5 2.5 Polyoxyethylene Sorbitan Monostearate 1.5 1.5 propylene glycol 4.0 4.0 Stearyl Glycyrrhetinate 0.2 0.2 petrolatum 2.0 2.0 antioxidant 0.5 0.5 Spices 0.1 0.1 antiseptic 0.5 0.5 Purified water balance balance

The compositions for external application for skin of Example 1 and Comparative Example 1 were applied to the skin and then evaluated by measuring transepidermal water loss (TEWL).

Specifically, two areas of 1.5 cm × 1.5 cm were set on the inner side of the forearm of the subject for 10 adults between the ages of 30 and 43, and the external skin composition of Example 1 and Comparative Example 1 was used as the test site. tested using

First, after measuring the transepidermal water loss of the test site using a transepidermal water loss measuring device (Vapometer, Delfin Technologies, Finland), the stratum corneum of the test site is removed through tape stripping to artificially damage the skin barrier made it After the skin barrier was damaged, the external skin composition of Example 1 and Comparative Example 1 was applied to the test site twice a day, and the amount of transepidermal water loss was measured after 1, 3, and 5 days, and the results were also shown. 3.

3 is a graph showing the degree of change in transepidermal water loss when the compositions for external application for skin according to Example 1 and Comparative Example 1 were applied.

Referring to FIG. 3, when the composition for external application for skin of Example 1 containing the callus extract of ginseng ginseng was applied according to one embodiment, the transepidermal water loss value was reduced compared to Comparative Example 1, indicating that the skin barrier improvement effect was excellent. .

Hereinafter, based on the results of the above experimental examples, various formulation examples using the callus extract of ginseng ginseng are formulated and presented. However, these formulation examples are for explaining one embodiment, and it is obvious to those skilled in the art that formulations in one embodiment are not limited to these formulation examples.

Formulation Example 1: Flexible skin toner

ingredient name Content (% by weight) Ginseng Callus Extract 0.5 glycerin 5.0 1,3-butylene glycol 3.0 ethanol 3.0 Polyoxyethylene nonylphenyl ether 0.5 Spices 0.1 antiseptic 0.5 Purified water balance

Formulation Example 2: Astringent

ingredient name Content (% by weight) Ginseng Callus Extract 0.5 glycerin 3.0 Citric Acid 0.1 ethanol 5.0 Polyoxyethylene oleyl ether 1.0 Sorbitol 2.0 Spices 0.1 antiseptic 0.5 Purified water balance

Formulation Example 3: Lotion (emulsion)

ingredient name Content (% by weight) Ginseng Callus Extract 0.5 glycerin 5.0 1,3-butylene glycol 8.0 squalane 10.0 Polyoxyethylene sorbitan monooleate 2.0 triethanolamine 1.5 Glyceryl Stearate 0.5 Stearyl Glycyrrhetinate 0.2 carboxyvinyl polymer 0.1 arginine 0.1 Spices 0.1 antiseptic 0.5 Purified water balance

Formulation Example 4: Cream

ingredient name Content (% by weight) Ginseng Callus Extract 0.5 glycerin 5.0 Stearic Acid 8.0 squalane 5.0 Self-emulsifying glycerin monostearate 2.5 Polyoxyethylene Sorbitan Monostearate 1.5 propylene glycol 4.0 Stearyl Glycyrrhetinate 0.2 petrolatum 2.0 antioxidant 0.5 Spices 0.1 antiseptic 0.5 Purified water balance

Formulation Example 5: Ointment

ingredient name Content (% by weight) Ginseng Callus Extract 0.5 Cetearyl Alcohol 2.0 Self-emulsifying monostearic acid 2.0 Stearic Acid 1.0 beeswax 4.0 squalane 7.0 monostearin glycerin 3.0 Sorbitan Monostearate 1.0 Polysorbate 80 3.0 glycerin 5.0 propylene glycol 4.0 Spices 0.1 petrolatum balance

As shown in Tables 5 to 9, according to one embodiment, the composition for external application for skin, in which various formulations are prepared using the extract of ginseng ginseng callus, can exhibit excellent effects on skin anti-inflammatory, promotion of ceramide biosynthesis, improvement of skin barrier, and skin moisturizing.

Although the preferred embodiments of the present invention have been described above, the present invention is not limited thereto, and it is possible to carry out various modifications within the scope of the claims and the detailed description of the invention and the accompanying drawings, and this is also the present invention. It goes without saying that it falls within the scope of the invention.

Claims (9)

It contains the callus extract of Scrophularia takesimensis Nakai extracted by hot water extraction as an active ingredient,
The concentration of the ginseng callus extract is 0.4 μg / mL to 400 μg / mL,
The serine palmitoyl transferase (SPT), ceramide synthase (CerS), or a combination thereof activating the serine palmitoyl transferase (SPT), or a composition for external application for skin. In paragraph 1,
The ginseng callus extract is obtained by extracting ginseng callus,
The ginseng callus is an external composition for skin that is obtained by inducing by culturing the leaves, stems, roots, flowers, or a combination thereof of ginseng in an induction medium. delete In paragraph 1,
The ginseng callus extract is contained in an amount of 0.0001% to 10% by weight based on the total amount of the composition for external application for skin. In paragraph 1,
The ginseng callus extract is a composition for external application for skin that inhibits the expression of inflammatory cytokines. In paragraph 1,
The extract for external application for skin is to promote ceramide biosynthesis. delete In paragraph 1,
The composition for external application for skin is a composition for external application for skin that is for improving the skin barrier, for moisturizing the skin, for relieving skin inflammation, or a combination thereof. In paragraph 1,
The composition for external application for skin is a cosmetic composition, a hair composition for improving the scalp, a pharmaceutical composition for improving the skin barrier and moisturizing the skin, or a combination thereof,
The pharmaceutical composition is suitable for dermatitis, prickly psoriasis, psoriasis, frostbite, acne, lichen, eczema, eczema, dermatitis, prurigo, psoriasis, toxic rash, psoriasis, psoriasis, palmar pustulosis, erythromatosis pilaris, and Sibert's rosea nonglycaemia. , erythema, erythematosus, discoid lupus erythematosus, systemic lupus erythematosus, pemphigus, pemphigoid, alopecia areata, vitiligo vulgaris, sarcoidosis, cutaneous amyloidosis, keloids, hypertrophic half-wounds, cuts, bedsores, skin ulcers , A composition for external application for skin that is for diseases of hair loss, wool or hair growth.

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