KR102397975B1 - External composition for moisturizing of skin comprising kirenol - Google Patents

Abstract

The present invention relates to a composition for external application for skin containing kylenol as an active ingredient, and more particularly, to a composition for external application for skin moisturizing or antioxidation containing kylenol as an active ingredient. The composition for external application for skin containing kylenol as an active ingredient according to the present invention promotes hyaluronic acid production and expression of synthetic enzymes to promote skin moisturizing, and exhibits excellent antioxidant effect by scavenging free radicals, which is effective for functional external composition for skin It can be used as an active ingredient.

Description

TECHNICAL FIELD [0001] External composition for moisturizing of skin comprising kirenol

The present invention relates to a composition for external application for skin containing kylenol as an active ingredient, and more particularly, to a composition for external application for skin moisturizing or antioxidation containing kylenol as an active ingredient.

Hyaluronic acid (hyaluronic acid) is a type of glycosaminoglycans (glycosaminoglycans), glucuronic acid and N-acetyl glucosamine residues are repeatedly linked to the chain-shaped polymer polysaccharide material. It combines with a large amount of water to form a gel, so it has high viscosity and elasticity. As a major component of the extracellular matrix, hyaluronic acid is reported to be involved not only in water retention, maintenance of intercellular spacing, storage and diffusion of cell growth factors and nutrients, but also in cell division, differentiation, and migration. there is.

In the skin, it has been reported that more than 50% of hyaluronic acid present in the body of a mammal is distributed in the skin, particularly the intercellular gap of the epidermis and the connective tissue of the dermis, and this hyaluronic acid is mainly synthesized by keratinocytes and fibroblasts It is known that It has been reported that the amount of hyaluronic acid in human skin decreases with aging, and the decrease in hyaluronic acid in the skin is considered to be one of the direct causes of the decrease in skin elasticity or moisture content according to aging (Biochem Biophys Acta 279, 265-275). ; Carbohydr Res 159, 127-136; Int J Dermatol 33, 119-122). Also, hyaluronic acid is known to be involved in maintaining the structure of the stratum corneum and maintaining the skin barrier function (J Cosmet Dermatol. 2007 Jun, 6(2), 75-82).

However, hyaluronic acid having the above effects is not well absorbed by the skin due to its high molecular weight. In addition, the method of injecting hyaluronic acid into the skin is currently being treated, A method of increasing hyaluronic acid synthesis in skin cells is more effective. Therefore, research on a method for increasing the production of hyaluronic acid in the human body is being actively conducted, but remarkable research results are not yet known.

On the other hand, active oxygen introduced from outside the living body or generated in the living body accelerates aging of the living body or causes many problems, such as cancer. Therefore, a lot of development and research on antioxidants that inhibit oxidation by active oxygen are being made. Antioxidants are widely distributed in animals and plants, and many phenolic compounds, flavonoids, tocopherols, vitamin C, and selenium are known in fruits and vegetables. However, antioxidant substances present in nature cannot be expected to have a substantially sufficient effect when applied to the skin. Therefore, although synthetic antioxidants with excellent antioxidant power and low price are widely used, their use is limited due to safety concerns such as side effects in the human body.

Therefore, there is an urgent need for the development of functional ingredients that are safe to the body and stable in active ingredients, and above all, have excellent skin moisturizing or antioxidant activity than existing functional substances.

The problem to be solved by the present invention is to provide a composition for external application for skin moisturizing the skin comprising kylenol as an active ingredient.

In addition, another problem to be solved by the present invention is to provide an antioxidant composition for external application for skin containing kylenol as an active ingredient.

In order to achieve the above technical object, the present invention provides a composition for external application for skin moisturizing comprising kirenol represented by the following formula (1) as an active ingredient. In addition, the present invention provides an antioxidant composition for external application for skin containing kylenol represented by the following formula (1) as an active ingredient.

[Formula 1]

Hereinafter, the configuration of the present invention will be described in detail.

In order for a skin condition improving ingredient to exhibit an excellent effect when applied to the actual skin, it must exhibit high skin condition improvement activity at a low concentration and have excellent ability to penetrate and absorb the skin. In addition, it is desirable that it has low volatility so that it can stay for a sufficient time to exhibit the skin condition improvement effect, the active ingredient is stably maintained in the composition or on the skin, easy to formulate into cosmetics, and safe for the skin.

However, among known components, components that satisfy all of the above properties are rare. For example, some skin condition improving ingredients have excellent skin condition improvement activity even at low concentrations during in vitro experiments, but their ability to penetrate and absorb the skin is poor, making it difficult to apply to actual skin. Another functional ingredient is difficult to formulate as a cosmetic because of its low hydrophilicity, or when exposed to heat, light, or oxygen, the active ingredient is decomposed or transformed into another compound, and the effect is already lost before application to the skin.

As a result of intensive research efforts to overcome the problems of the prior art, the present inventors have found that a composition for external application for skin containing kirenol promotes the production of hyaluronic acid in keratinocytes, and hyaluronic acid synthase (hyaluronic acid synthase, HAS), and was found to increase the synthesis of hyaluronic acid in the body. In addition, it was found that it has an excellent antioxidant effect by scavenging free radicals, thereby completing the present invention.

The kirenol used in the present invention is Siegesbeckia glabrescens ) is one of the active ingredients of the extract.

As can be seen in the following examples, kylenol exhibits an exceptionally excellent hyaluronic acid production promoting effect or antioxidant effect at a low concentration. Therefore, kylenol can be used as an active ingredient in a composition for external application for skin for moisturizing or antioxidant effect.

The present invention provides a composition for external application for skin moisturizing the skin comprising kylenol as an active ingredient. In addition, the composition for external application for skin has excellent skin regeneration ability and may be used for wound healing on the skin surface.

As used herein, the term "skin moisturizing" refers to maintaining homeostasis of a living body by appropriately controlling moisture loss (water evaporation), etc., and when hyaluronic acid or collagen in the skin is reduced, moisture is lost, If increased, moisture may increase.

The composition for external application for skin moisturizing the skin comprising kylenol as an active ingredient of the present invention is characterized in that it promotes the production of hyaluronic acid. In addition, the composition is characterized in that it increases the expression of hyaluronic acid synthase (hyaluronic 　 acid 　 synthase, 　HAS).

The present invention is kylenol It is based on the fact that it can increase the expression of hyaluronic acid protein in keratinocytes derived from the human body. In the present invention, an experiment was conducted to examine the hyaluronic acid production promoting effect of chilenol, and as shown in Examples 2 to 4 of the present invention, when chilenol was treated, the concentration of hyaluronic acid was increased, and also hyaluronic acid synthesis It was confirmed that the expression of the protein of hyaluronic acid was increased by the gene expression of the enzyme.

As used herein, the term "hyaluronic acid" is a type of glycosaminoglycans and refers to a chain-shaped polymeric polysaccharide material in which glucuronic acid and N-acetylglucosamine residues are repeatedly linked.

In addition, the present invention provides an antioxidant composition for external application for skin comprising kylenol as an active ingredient. The composition for external application for skin as an antioxidant is characterized in that it exhibits an antioxidant effect by inhibiting the generation of radicals. The antioxidant composition for external application for skin can effectively remove active oxygen introduced from the outside of the body or generated in the body, and thus can be used for skin anti-aging or anti-cancer purposes.

In the present invention, an experiment was conducted to investigate the radical generation inhibitory effect of chilenol, and as shown in Example 5 of the present invention, when kylenol was treated, it exhibited a higher radical scavenging activity than ascorbic acid, and thus the antioxidant effect was very It was confirmed that it was excellent.

The term "antioxidant" effect of the present invention inhibits cell oxidation by highly reactive free radicals or reactive oxygen species (ROS) depending on intracellular metabolism or oxidative stress caused by the effect of ultraviolet rays This includes reducing damage to cells by removing free radicals or reactive oxygen species.

The composition for external application for skin moisturizing or antioxidant according to the present invention may be used as a cosmetic composition for improving skin condition or a pharmaceutical composition for preventing or treating skin diseases.

In the present invention, the composition for external application for skin contains kylenol in an amount of 0.00001 to 10% by weight, preferably 0.0001 to 7% by weight, more preferably 0.001 to 5% by weight, most preferably 0.01 to 3% by weight, based on the total weight of the composition. % can be included. At a concentration of less than 0.00001% by weight of the kylenol composition for external application for skin, it is difficult to obtain its effect, and at a concentration of more than 10% by weight, a problem may occur in formulation stability.

The kylenol of the present invention increases the protein expression of hyaluronic acid in the skin to promote production and has an antioxidant effect, so it can be usefully used as an active ingredient in various external preparations for skin. When the present invention is used as a composition for external application for skin, it can be used in any formulation applicable to the skin, such as liquid, oil, cream, ointment, stick, pack, pasta, or powder.

More specifically, the formulation of the composition for external application for skin is a solution, external ointment, cream, foam, nourishing lotion, softening lotion, perfume, pack, soft water, emulsion, makeup base, essence, soap, liquid detergent, bathing agent, sunscreen cream , sun oil, suspension, emulsion, paste, gel, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, patch, and spray, etc. and can be used

The composition for external application for skin containing kylenol according to the present invention is a cosmetically or pharmaceutically acceptable carrier, diluent, adjuvant, colorant, and stabilizer for easy use and handling within the scope that does not impair the achievement of the object of the present invention. , a flavoring agent, a surfactant, an oil, a moisturizing agent, alcohol, a thickener, an antioxidant, a pH adjusting agent, or a sunscreen agent and the like may be additionally contained.

The composition for external application for skin containing kylenol as an active ingredient according to the present invention promotes hyaluronic acid production and expression of synthetic enzymes to promote skin moisturizing, and exhibits excellent antioxidant effect by scavenging free radicals, which is effective for functional external composition for skin It can be used as an active ingredient.

Hereinafter, examples and the like will be described in detail to help the understanding of the present invention. However, the embodiments according to the present invention may be modified in various other forms, and the scope of the present invention should not be construed as being limited to the following examples. The embodiments of the present invention are provided to more completely explain the present invention to those of ordinary skill in the art.

preparation of materials

Cylenol used in the following examples was purchased from Shaanxi Zhujiang Biotechnology Co., Ltd. (陝西舟康生物有限公司) and used.

Example One: of kylenol Cytotoxicity assessment

To evaluate the cytotoxicity of kylenol, cell division changes of human keratinocytes (HaCaT) were measured using Cell Counting Kit-8 (Dojindo Lab, USA).

First, a uniform number of cells were spread on a 96-well plate, stabilized, and then treated with chilenol to 0.5 or 1 ppm, and further cultured for 24 hours. After 2 hours by treatment with CCK-8, the absorbance was measured at 450 nm. The relative cell viability of the cells treated with kylenol compared to the untreated group (control group) was analyzed as follows, and the results are shown in Table 1 below.

sample untreated group Glucosamine hydrochloride
1000 ppm kylenol
0.5 ppm kylenol
1 ppm Survival rate (%) 100 96 98 94

* number of repetitions n = 3

* Relative cell viability = (extract treated group)/(untreated group) X 100

As shown in Table 1, it was found that the number of cells was not significantly changed by kylenol treatment. This means that kylenol does not cause cytotoxicity.

Example 2: Human-derived keratinocytes using enzyme-linked immunosorbent assay (ELISA) to kylenol Increased hyaluronic acid concentration by

Human-derived keratinocytes were cultured in DMEM medium containing 10% fetal bovin serum at 1x10 ⁵ cells/ml, dispensed in a 24-well plate by 500 μl, and cultured for 18 hours. After washing twice with non-DMEM medium, kylenol was added at different concentrations (0.5 and 1 ppm). The same amount of ethanol was treated as a negative control, and 100 ppm of glucosamine hydrochloride (Sigma-aldrich), known to promote hyaluronic acid production, was treated as a positive control. After culturing for 24 hours, the cell culture medium was recovered and the hyaluronic acid concentration was measured using a Hyaluronan ELISA kit (R&D Systems), and it is shown in Table 2.

sample unprocessed
(negative control) Glucosamine hydrochloride
100 ppm kylenol
0.5 ppm kylenol
1 ppm hyaluronic acid
Relative concentration (%) 100 139 131 169

* number of repetitions n = 3

* Numericalized the hyaluronic acid concentration of the experimental group treated with chilenol based on the hyaluronic acid concentration of the negative control group (100%).

As a result, kylenol increased the production of hyaluronic acid in human-derived keratinocytes in a concentration-dependent manner.

Example 3: In human-derived keratinocytes to kylenol Increased expression of hyaluronic acid synthase (HAS) gene by

Human-derived keratinocytes were cultured in DMEM medium containing 10% fetal bovin serum at 1x10 ⁶ cells/ml, aliquoted at 3000 μl in a 6-well plate, and incubated for 18 hours. It was washed twice with non-DMEM medium and treated with 0.5 ppm and 1 ppm of kylenol for 24 hours. Cells were recovered, total RNA was extracted with RNeasy mini kit (Qiagen), quantified, and 1 μg of RNA was reverse transcribed using GeneAmp ^® RNA PCR kit (Applied Biosystems). Reverse transcription reaction was performed using Mycycler ^® PCR instrument (Biorad).

Then, quantitative real-time PCR was performed using 2 μl of the reverse transcription reaction solution and Taqman ^® probe (Invitrogen, USA, HS02511055_S1, HS03929097_g1) of HAS-2, HAS-3, and GAPDH (Glyceraldehyde-3-phosphate dehydrogenase). Real time PCR was performed using CFX96 Touch ^TM Real-Time PCR Detection System (Biorad). Experimental results obtained through real-time PCR were calculated by the method described in Livak KJ et al. (Methods, 2001, 25, 402-408) based on the intracellular control gene GAPDH.

In addition, the mRNA expression level of the kylenol-treated experimental group was quantified based on the mRNA expression level of the negative control group as a reference (100%), and it is shown in Table 3 below.

sample unprocessed
(negative control) Glucosamine hydrochloride
100 ppm kylenol
0.5 ppm kylenol
1 ppm HAS-2
Expression (%) 100 125 109 119 HAS-3
Expression (%) 100 130 106 125

* number of repetitions n = 3

As a result, expression of hyaluronic acid synthase, particularly HAS-2 and HAS-3, was increased in human-derived keratinocytes by treatment with kylenol.

Example 4: in human fibroblasts to kylenol Increased hyaluronic acid concentration by

Human Dermal Fibroblasts were cultured at 1x10 ⁵ cells/ml in DMEM medium containing 10% fetal bovin serum, dispensed 500 μl into a 24-well plate, and incubated for 18 hours, followed by culturing for 18 hours. Wash twice with DMEM-free medium. Cylenol was added at different concentrations (0.5 and 1 ppm). The same amount of ethanol was treated as a negative control, and 100 ppm of glucosamine hydrochloride (Sigma-aldrich), which is known to promote hyaluronic acid production, was treated as a positive control. After culturing for 24 hours, the cell culture medium was recovered and the hyaluronic acid concentration was measured using a Hyaluronan ELISA kit (R&D Systems), and it is shown in Table 4.

sample unprocessed
(negative control) Glucosamine hydrochloride
100 ppm kylenol
0.5 ppm kylenol
1 ppm hyaluronic acid
Relative concentration (%) 100 167 189 206

* number of repetitions n = 3

From the experimental results shown in Table 4, kylenol increased the production of hyaluronic acid in human-derived fibroblasts.

Example 5: to kylenol Antioxidant effect by - free radical scavenging rate

To confirm the antioxidant activity of kylenol, free radical scavenging activity was measured. Free radical scavenging ability was measured by the 1,1-diphenyl-2-picryl-hydrazyl (DPPH) method as follows (Blois, M.S. Nature 181, 1190, 1958). DPPH is a relatively stable free radical, and when it exists in a radical state, it exhibits a maximum absorption at 517 nm and loses absorption when it is eliminated. DPPH was used by Sigma, dissolved in methyl alcohol at a concentration of 0.15 mM. First, 100 μl of kylenol and ascorbic acid as a positive control were added to each well of a 96-well plate. 100 μl of DPPH solution was added thereto, left at room temperature for 30 minutes, and then absorbance at 517 nm was measured using a micro plate reader (BioTek EL-340). At this time, as a control, methyl alcohol was used instead of chilenol. The free radical scavenging activity effect was obtained with the absorbance measured for the experimental group and the control group, and the results are shown in Table 5 below.

Free radical removal rate (%) sample Free radical removal rate (%) kylenol 56.2 ascorbic acid 78.9 methyl alcohol 8.4

As can be seen from the results of Table 5, it can be seen that kylenol has a high activity compared to ascorbic acid, which is a known antioxidant, and thus has an antioxidant effect.

Example 6: to kylenol increase in skin moisture

In order to apply the effect of increasing the skin moisture content of the skin external preparation containing kylenol to human skin, it was prepared according to the composition of the nutrient cream of Preparation Example 1 below, and was administered to 20 healthy adults in a constant temperature and humidity room at 22°C and 50% relative humidity. A certain amount (2 mg/cm 2 ) was applied to the inside of the lower arm, and the moisture content of the skin was measured before application, 1 hour after application, and 2 hours after application. In the case of the control group, a nourishing cream containing the same amount of ethanol was prepared and applied in the same way instead of kylenol, and normal skin without any treatment was used as the untreated group. The water retention capacity was measured by measuring the electrical conductivity of the skin surface using a Corneometer (Courage + Khazaka, GmbH, Germany). The results are shown in Table 6 below.

unit (corneometer units) Preparation Example 1
nourishing cream control
nourishing cream untreated group the battlefield 53 55 57 1 hour after application 84 79 55 2 hours after application 72 65 54

* n = 20

As a result, the nourishing cream containing kylenol increased the amount of moisture in human skin, and showed excellent moisture retention ability even after 2 hours of application.

Through this, it was confirmed that kylenol has a very excellent effect of increasing the amount of skin moisture and retaining moisture, so it can be usefully used for moisturizing the skin.

Based on the effect test results as described above, the preparation example of the composition according to the present invention will be described in detail. However, the following preparation examples are only examples of the composition of the present invention, it will be apparent to those of ordinary skill in the art that the scope of the composition of the present invention is not limited only to the following formulations. Hereinafter, formulation examples of external preparations for skin containing kylenol are shown by preparation examples.

production example 1: Preparation of nourishing cream

Nutrient cream containing kylenol as an active ingredient with the composition shown in Table 7 below was prepared by a conventional method used in the cosmetic field.

Ingredient name content (wt%) kylenol 1.0 beta-1,3-glucan 5.0 beeswax 10.0 Polysorbate 60 1.5 sebum 60 hydrogenated castor oil 2.0 Sorbitan sesquioleate 0.5 liquid paraffin 10.0 squalane 5.0 Caprylic/Capric Triglycerides 5.0 glycerin 5.0 butylene glycol 3.0 propylene glycol 3.0 triethanolamine 0.2 antiseptic 0.05 pigment 0.05 Spices 0.05 Purified water To 100

From the above description, those skilled in the art to which the present invention pertains will understand that the present invention may be embodied in other specific forms without changing the technical spirit or essential characteristics thereof. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention, rather than the above detailed description, all changes or modifications derived from the meaning and scope of the claims to be described later and their equivalents.

Claims (5)

A composition for external application for skin moisturizing by enhancing hyaluronic acid comprising kirenol as an active ingredient. delete delete The composition for external application for skin according to claim 1, wherein the composition comprises 0.00001 to 10% by weight of kylenol based on the total weight of the composition. According to claim 1, wherein the formulation of the composition is a solution, external ointment, cream, foam, nourishing lotion, softening lotion, perfume, pack, soft water, emulsion, makeup base, essence, soap, liquid detergent, bathing agent, sunscreen cream , sun oil, suspension, emulsion, paste, gel, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, patch, and spray A composition for external application to the skin.