KR20220011402A - Composition for restoring abnormal differentiation of skin keratin - Google Patents

Abstract

The present disclosure relates to a composition for normalizing abnormal differentiation of dead skin cells. The composition comprises an extract of Artemisia Capillaris as an active ingredient. The composition recovers expression of keratin 15 and transglutaminase 3 of keratinocyte which is deformed by fine dust and normalizes abnormal differentiation of dead skin cells.

Description

Composition for normalizing differentiation abnormality of skin keratin {COMPOSITION FOR RESTORING ABNORMAL DIFFERENTIATION OF SKIN KERATIN}

The present specification relates to a composition for normalizing the differentiation abnormality of skin keratin.

Recently, as an environmental issue, interest in fine dust is increasing and the pollution market is expanding. Fine dust penetrates the human body better because it has smaller particles than general yellow dust, and it is harmful to the human body because it contains more harmful substances such as heavy metals, nitrates, ammonium, and sulfates. When fine dust enters our body, the cells in charge of the body's immunity act to remove the dust. As a side effect at this time, inflammation is accompanied, and in addition, the World Health Organization classifies fine dust as a group 1 carcinogen directly related to cancer.

Fine dust penetrates not only the respiratory tract but also the skin and is expected to damage the skin barrier or cause skin problems, but research on the mechanism and target of how it negatively affects the skin is insufficient.

Accordingly, it is necessary to develop an anti-pollution material capable of alleviating damage to the skin barrier caused by fine dust.

Korean Patent Publication No. 10-2019-0033256

In one aspect, the present invention is to provide a composition for normalizing the differentiation abnormality of the skin keratin.

In one aspect, the present invention provides a composition for normalizing the differentiation of keratin of the skin comprising an extract of Artemisia wormwood as an active ingredient.

In an exemplary embodiment, the differentiation abnormality of the keratin of the skin may be due to fine dust.

In an exemplary embodiment, the Mugwort extract may be an extract of water, an organic solvent, or a mixture thereof.

In an exemplary embodiment, the wormwood extract may be administered at a dosage of 5 to 50 mg/kg/day.

In an exemplary embodiment, the fine dust may include one or more of arsenic, cadmium, lead, and nickel.

In an exemplary embodiment, the fine dust may have a particle size of PM 10 or less.

In an exemplary embodiment, the active ingredient may increase the expression of keratin 15 (Keratin 15, KRT15).

In an exemplary embodiment, the active ingredient may reduce the expression of transglutaminase 3 (TGM3).

In an exemplary embodiment, the composition may be a composition for external application to the skin.

In an exemplary embodiment, the composition may be a food, cosmetic or pharmaceutical composition.

In one aspect, the composition provided by the present invention can normalize the differentiation abnormality of skin keratin.

In one aspect, the composition provided by the present invention can increase the expression of keratin 15 reduced by fine dust.

In one aspect, the composition provided by the present invention can reduce the expression of transglutaminase 3 increased by fine dust.

1 is a graph showing the cytotoxicity according to the concentration of Mugwort extract.
Figure 2 is a graph showing the expression of KRT15 according to the treatment of Mugwort extract.
3 is a graph showing the expression of TGM3 according to the treatment of Mugwort extract.

Hereinafter, the present invention will be described in detail.

In one aspect, the present invention provides a composition for normalizing differentiation of keratin of the skin comprising an extract of Artemisia capillaris as an active ingredient.

In the present specification, Artemisia capillaris is a perennial herb that belongs to the Asteraceae and grows wild in Korea. It does not die even in winter and sprouts again from the stem the following year, so it is called Artemisia capillaris. The main components of wormwood are 6,7-dimethoxycoumarin, chlorogenic acid, caffeic acid, β-pinene, and capillin. (capillin), stearic acid, palmitic acid, oleic acid, etc. have been reported.

In one aspect, the wormwood extract used as an active ingredient of the present invention can be prepared by extracting wormwood with water or an organic solvent by a method known in the art. The organic solvent may be at least one selected from the group consisting of ethanol, methanol, butanol, ether, ethyl acetate and chloroform, or a mixed solvent of these organic solvents and water may be used. Preferably, 80% ethanol may be used.

In one embodiment, the abnormal differentiation of the keratin of the skin may be due to fine dust.

In one embodiment, the wormwood extract may be administered at a dosage of 5 to 50 mg/kg/day. When the dosage of the wormwood extract is less than 5 mg/kg/day, it may be difficult to exhibit the effect according to the present invention, and when it exceeds 50 mg/kg/day, it may cause irritation to the skin. Specifically, the active ingredient of the composition is 5 mg/kg/day or more, 5.1 mg/kg/day or more, 5.2 mg/kg/day or more, 5.3 mg/kg/day or more, 5.4 mg/kg/day or more, 5.5 mg/kg/day or more, 5.6 mg/kg/day or more, 5.7 mg/kg/day or more, 5.8 mg/kg/day or more, 5.9 mg/kg/day or more, 6 mg/kg/day or more, 6.5 mg/ ≥ kg/day, ≥ 7 mg/kg/day, ≥ 7.5 mg/kg/day, ≥ 8 mg/kg/day, ≥ 9 mg/kg/day, ≥ 10 mg/kg/day, ≥ 11 mg/kg/ ≥ 12 mg/kg/day, ≥ 13 mg/kg/day, ≥ 14 mg/kg/day, ≥ 15 mg/kg/day, ≥ 16 mg/kg/day, ≥ 17 mg/kg/day, 18 mg/kg/day or more, 19 mg/kg/day or more, or 20 mg/kg/day or more, but 50 mg/kg/day or less, 49 mg/kg/day or less, 48 mg/kg/day or less, 47 less than or equal to mg/kg/day, less than or equal to 46 mg/kg/day, less than or equal to 45 mg/kg/day, less than or equal to 44 mg/kg/day, less than or equal to 43 mg/kg/day, less than or equal to 42 mg/kg/day, less than or equal to 41 mg/ kg/day or less, 40 mg/kg/day or less, 39 mg/kg/day or less, 38 mg/kg/day or less, 37 mg/kg/day or less, 35 mg/kg/day or less, 33 mg/kg/day It may be administered at a dosage of no more than 30 mg/kg/day, no more than 27 mg/kg/day, or no more than 25 mg/kg/day.

As used herein, the term “fine dust” refers to particulate matter that is invisible to our eyes and floats or scatters in the atmosphere for a long time. Specifically, the particle size of the fine dust may be 10 μm or less (PM 10), more specifically, 2.5 μm or less (PM 2.5). In particular, particulate matter having a particle diameter of 2.5 μm or less (PM 2.5) is referred to as “ultra-fine dust”, and in this specification, “fine dust” is intended to include “ultra-fine dust”.

In one aspect, the fine dust may include at least one of arsenic, cadmium, lead, and nickel. The heavy metal components such as arsenic, cadmium, lead, and nickel may be included in fine dust to attack the skin and cause abnormal differentiation of keratinocytes. In one embodiment, the fine dust may include one or more of arsenic, cadmium, lead, and nickel.

In one embodiment, the active ingredient may increase the expression of keratin 15 (Keratin 15, KRT15) of keratinocytes reduced by fine dust. Keratin 15 is a protein that plays an important role in maintaining epithelial stem cells, and is known to be necessary for skin regeneration and skin repair to occur properly. Keratin 15 regulates the differentiation of keratinocytes, and when keratinocytes are exposed to fine dust, the expression of keratin 15 is reduced.

In one embodiment, the active ingredient may reduce the expression of transglutaminase 3 (TGM3) in keratinocytes increased by fine dust. Transglutaminase 3 is a protein that regulates epidermal differentiation, and when keratinocytes are exposed to fine dust, the expression of transglutaminase 3 is increased.

In one embodiment, the route of administration of the composition is not limited, but may be preferably transdermally or externally applied to the skin.

In one embodiment, the composition may be a cosmetic composition. The cosmetic composition is, for example, softening lotion, astringent lotion, nourishing lotion, nourishing cream, massage cream, eye cream, eye essence, essence, cleansing cream, cleansing lotion, cleansing foam, cleansing water, pack, powder, body lotion , body cream, body essence, body cleaner, hair dye, shampoo, conditioner, hair dressing, hair conditioner, ointment, gel, cream, patch, spray, powder, and skin adhesion type, but are not limited thereto.

In addition, in each formulation, other components other than the above essential components can be appropriately selected and formulated by those skilled in the art without difficulty depending on the type or purpose of use of other external preparations.

The cosmetic composition may be provided in any formulation suitable for topical application. For example, it may be provided in the form of a solution, an emulsion obtained by dispersing an oil phase in an aqueous phase, an emulsion obtained by dispersing an aqueous phase in an oil phase, a suspension, a solid, a gel, a powder, a paste, a microneedle, a foam, or an aerosol composition. have. Compositions of such formulations can be prepared according to conventional methods in the art.

The cosmetic composition according to the present specification may further include functional additives and components included in general cosmetic compositions in addition to the compounds of the present specification. The functional additive may include a component selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingolipids, and seaweed extract. The cosmetic composition according to the present specification may include other ingredients capable of giving a synergistic effect to the main effect, preferably within a range that does not impair the main effect. In addition, the cosmetic composition according to the present specification may further include a moisturizing agent, an emollient, a surfactant, a UV absorber, a preservative, a bactericide, an antioxidant, a pH adjuster, an organic and inorganic pigment, a fragrance, a cooling agent or a limiting agent. The blending amount of the component can be easily selected by those skilled in the art within the range that does not impair the purpose and effect of the present specification, and the blending amount may be 0.001 to 10% by weight, specifically 0.01 to 3% by weight, based on the total weight of the composition. have.

In one embodiment, the composition may be a food composition. The formulation of the food composition is not particularly limited, but, for example, may be formulated as tablets, granules, pills, powders, liquids such as drinks, caramel, gels, bars, tea bags, and the like. In addition to the active ingredient, the food composition of each formulation can be appropriately selected by those skilled in the art without difficulty depending on the formulation or purpose of use in addition to the active ingredient, and a synergistic effect may occur when applied simultaneously with other raw materials.

The food composition according to one embodiment includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavoring agents, coloring agents and enhancers (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the food compositions according to an embodiment may include natural fruit juice and pulp for the production of fruit juice beverages and vegetable beverages. These components may be used independently or in combination. The proportion of these additives is not so important, but is generally included in the range of 0 to about 50 parts by weight per 100 parts by weight of the composition according to one embodiment.

In one embodiment, the composition may be a pharmaceutical composition. The pharmaceutical composition may be administered orally, parenterally, rectally, topically, transdermally, intravenously, intramuscularly, intraperitoneally, subcutaneously, or the like. Formulations for oral administration may be tablets, pills, soft and hard capsules, granules, powders, fine granules, liquids, emulsions, or pellets, but are limited thereto it is not Formulations for parenteral administration may be solutions, suspensions, emulsions, gels, injections, drops, suppositories, patches, or sprays, but is not limited thereto. The formulations can be easily prepared according to conventional methods in the art, and surfactants, excipients, wettable powders, emulsification accelerators, suspending agents, salts or buffers for osmotic pressure control, colorants, spices, stabilizers, preservatives, preservatives or Other commercially available adjuvants may be further included.

Hereinafter, the present invention will be described in more detail through examples and the like. These examples are only for illustrating the present invention, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not to be construed as being limited by these examples.

[Example 1] Preparation of wormwood extract

After washing and drying 1 kg of wormwood (Jeju Oedo), 30L of water was added and extracted for 24 hours. After filtration using a 250 mesh filter, the obtained extract was concentrated and dried to obtain a wormwood extract.

[Experimental Example 1] Culturing of keratinocytes

Keratinocytes were cultured in the (Normal Human Epidermal neonatal keratinocytes-, LONZA社, 192906) a CO ₂ incubator (CO ₂ incubator) at 37 ℃, 5% CO ₂ condition. As a medium, ^{Supplement (LONZA, 192060) was added to KGM-Gold TM} Keratinocyte Growth Medium, and Accutase (Merck Millipore, SCR005) was used for cell passage.

[Experimental Example 2] Cytotoxicity measurement experiment

The keratinocytes of Experimental Example 1 were inoculated in a 96-well plate at 20000 cells/well (cells/well), and then cultured at _{37° C. and 5% CO 2 conditions.} When the cultured keratinocytes subconfuent, the Mugwort extract of Example 1 was treated by concentration and cultured at _{37° C. and 5% CO 2 conditions for 24 hours.} An untreated group was used as a control. After treatment with EZ-Cytox (Daeil Labservice) for 1 hour, absorbance was measured at 450 nm using a microplate reader, and the results are shown in FIG. 1 .

As can be seen in FIG. 1 , the Mugwort extract did not show cytotoxicity even at 50 ppm.

[Experimental Example 3] KRT15 and TGM3 expression level measurement experiment through qPCR

1. Treatment of fine dust and substances in keratinocytes

Fine dust was collected at a PM 2.5 level by installing a high volume sampler on the roof of the Amorepacific R&D Center. A PTFE Teflon filter was used, and it was collected for 24 hours at a flow rate of ^{1.13 m 3 /min.}

In a 12 well plate, the keratinocytes of Experimental Example 1 were inoculated at 3 × 10 ⁵ cells/well (cells/well), and fine dust (100 μg/ml) and mugwort of Example 1 were inoculated when the keratinocytes were subconfused. The extracts were treated by concentration and incubated for 24 hours.

2. qPCR Quantification

2-1. RNA extraction

The culture medium was removed from the 12 well plate treated with the material, and the cells were washed with 2 ml of phosphate buffer (Phosphate Buffered Saline, PBS), and then RNA was isolated from the cells using Trizol reagent (Invitrogen).

2-2. cDNA synthesis

cDNA was synthesized from the isolated RNA using a reverse transcription kit (Invitrogen, Superscript III).

2-3. Perform qPCR

The synthesized cDNA was quantitatively analyzed through quantitative polymerase chain reaction (qPCR) to measure the expression levels of KRT15 and TGM3, and the results are shown in FIGS. 2 and 3 . The TaqMan probe used to measure the expression level of KRT15 was Hs00951967_g1, the TaqMan probe used to measure the expression level of TMG3 was Hs00162752_m1, and their expression level was corrected with RPL13A (Hs04194366_g1).

As shown in FIG. 2 , when fine dust was treated with keratinocytes, the expression of KRT15 was significantly reduced compared to the control group not treated with fine dust. It was confirmed that the expression of KRT15 changed by dust was restored.

In addition, as shown in FIG. 3 , when fine dust was treated with keratinocytes, the expression of TGM3 was significantly increased compared to the control group not treated with fine dust, but when fine dust and Mugwort extract were treated together, the expression of TGM3 decreased. Thus, it was confirmed that the expression of TGM3 changed by fine dust was restored.

[Formulation Example 1] Lotion

A lotion was prepared in a conventional manner according to the composition shown in Table 1 below.

ingredient content wormwood extract 5 g L-ascorbic acid-2-magnesium phosphate salt 1.5 g Collagen 1.5 g sodium citrate 0.15 g citric acid 0.08 g Licorice Extract 0.3 g 1,3-butylene glycol 4.5 g Purified water 137 g

[Formulation Example 2] Cream

Creams were prepared in a conventional manner according to the composition shown in Table 2 below.

ingredient content wormwood extract 4.5 g Polyethylene glycol monostearate 2.5 g Self-emulsifying glycerin monostearate 6.5 g cetyl alcohol 5 g squalene 7.5 g tri2-glyceryl ethyl henate 7.5 g sphingoglycolipids 1 g 1,3-butylene glycol 8 g Purified water 60 g

[Formulation Example 3] Pack

A pack was prepared in a conventional manner according to the composition shown in Table 3 below.

ingredient content wormwood extract 1.5 g polyvinyl alcohol 13 g L-ascorbic acid-2-magnesium phosphate salt 1 g Lauroylhydroxyproline 1 g Collagen 2 g 1,3-butylene glycol 3 g ethanol 5 g Purified water 70 g

[Formulation Example 4] Soap

Soap was prepared in a conventional manner according to the composition shown in Table 4 below.

ingredient content wormwood extract 4.5 g titanium dioxide 0.2 g polyethylene glycol 0.8 g glycerin 0.5 g Ethylenediaminetetraacetic acid 0.05 g salt 1 g pigment 0.2 g soapy scent 0.2 g soap base 93 g

[Formulation Example 5] Cosmetic liquid formulation

According to the composition shown in Table 5 below, a cosmetic liquid formulation was prepared in a conventional manner.

ingredient content wormwood extract 5 g Hydroxyethylene Cellulose 30 g xanthan gum 5 g 1,3-butylene glycol 15 g glycerin 10 g Sodium Hyaluronate 12 g Purified water 173 g

[Formulation Example 6] Soft capsules

A soft capsule was prepared by mixing 150 mg of wormwood extract, 160 mg of L-carnitine, 200 mg of soybean oil, 2 mg of palm oil, 8 mg of hydrogenated vegetable oil, 4 mg of yellow wax and 6 mg of lecithin, and filling one capsule according to a conventional method.

[Formulation Example 7] Tablet

After mixing 120 mg of wormwood extract, 200 mg of galactooligosaccharide, 60 mg of lactose, and 140 mg of maltose, granulation using a fluidized bed dryer, 6 mg of sugar ester was added thereto, and the tablets were tableted with a tableting machine.

[Formulation Example 8] Granules

200 mg of Artemisia Artemisia extract, 250 mg of anhydrous crystalline glucose and 550 mg of starch were mixed, formed into granules using a fluidized bed granulator, and then filled in a bag to prepare granules.

[Formulation Example 9] Drink formulation

After mixing 300 mg of mugwort extract, 10 g of glucose, 0.6 g of citric acid, and 25 g of liquid oligosaccharide, add 300 ml of purified water and fill each bottle by 200 ml. After filling the bottle, it was sterilized at 130° C. for 4 to 5 seconds to prepare a drink beverage.

Claims (12)

A composition for normalizing differentiation of keratin of the skin comprising an extract of Artemisia capillaris as an active ingredient. The method of claim 1,
The abnormal differentiation of the skin keratin is due to fine dust, the composition. The method of claim 1,
The composition, wherein the wormwood extract is an extract of water, an organic solvent, or a mixed solvent thereof. The method of claim 1,
The composition composition wherein the wormwood extract is administered at a dosage of 5 to 50 mg/kg/day. 3. The method of claim 2,
The fine dust is a composition comprising at least one of arsenic, cadmium, lead, and nickel. 3. The method of claim 2,
The fine dust is a composition having a particle size of PM 10 or less. The method of claim 1,
The active ingredient is to increase the expression of keratin 15 (Keratin 15), the composition. The method of claim 1,
The active ingredient is to reduce the expression of transglutaminase 3 (Transglutaminase 3), the composition. 9. The method according to any one of claims 1 to 8,
The composition is a composition for external application to the skin, the composition. 9. The method according to any one of claims 1 to 8,
The composition is a food composition. 9. The method according to any one of claims 1 to 8,
The composition is a cosmetic composition, composition. 9. The method according to any one of claims 1 to 8,
The composition is a pharmaceutical composition.

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