KR20170030831A - External composition for moisturizing of skin comprising kirenol - Google Patents

Abstract

The present invention relates to a composition for external application for skin comprising kirenol as an active ingredient, and more specifically to a skin external application composition for skin moisturizing or antioxidation, which contains a kirenol as an active ingredient. The composition for external application for skin, which comprises the cholineol according to the present invention as an active ingredient, accelerates hyaluronic acid production and expression of a synthetic enzyme, thereby promoting skin moisturization and eliminating free radicals to exhibit an excellent antioxidative effect. It can be used as an active ingredient.

Description

[0001] The present invention relates to an external composition for moisturizing a skin comprising kirenol,

The present invention relates to a composition for external application for skin comprising kirenol as an active ingredient, and more specifically to a skin external application composition for skin moisturizing or antioxidation, which contains a kirenol as an active ingredient.

Hyaluronic acid is a kind of glycosaminoglycans, and is a chain-like polymer polysaccharide substance in which glucuronic acid and N-acetylglucosamine residues are repeatedly connected. It has high viscosity and elasticity because it has a property of making gel by combining with a large amount of water. Hyaluronic acid is a major component of the extracellular matrix and has been reported to be involved not only in the retention of water content, intercellular spacing, storage and spread of cell growth factors and nutrients, but also in cell division, differentiation and migration have.

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

However, the hyaluronic acid having the above effect has a large molecular weight and is not well absorbed into the skin. In addition, although a method of injecting hyaluronic acid into the skin is currently performed, A method of increasing hyaluronic acid synthesis in skin cells is more effective. Therefore, studies on a method for increasing the production of hyaluronic acid in the human body are actively conducted, but the remarkable results of the study are not yet known.

On the other hand, active oxygen introduced from the outside of the living body or generated in the living body causes many problems such as promoting aging of the living body, cancer, and the like. Therefore, the development and research of antioxidants that inhibit oxidation by active oxygen have been performed. Antioxidants are widely distributed in copper and plants, and many phenolic compounds, flavonoids, tocopherols, vitamin C, and selenium are known in fruits and vegetables. However, the antioxidant substances present in nature can not be expected to have practically sufficient effects in skin application. Therefore, although synthetic antioxidants having excellent antioxidant ability and low cost are widely used, their use is restricted due to safety concerns such as human side effects.

Therefore, it is urgently required to develop a functional ingredient having skin moisturizing or antioxidant activity which is safe in living body, stable in its active ingredient and above all, more effective than existing functional substances.

The object of the present invention is to provide a skin external application composition for moisturizing skin comprising as an active ingredient, a quinolone.

Another object of the present invention is to provide an antioxidant composition for external application for skin, which comprises choline as an active ingredient.

In order to accomplish the above object, the present invention provides a skin external composition for skin moisturizing comprising Kirenol represented by Chemical Formula 1 as an active ingredient. The present invention also provides an antioxidant composition for external application for skin, which comprises, as an active ingredient, a quinolone represented by the following formula (1).

[Chemical Formula 1]

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

In order to exert an excellent effect when the skin condition improving ingredient is applied to the actual skin, it exhibits a highly active skin condition improving activity at a low concentration and should be excellent in the ability to permeate and absorb the skin. In addition, it is preferable that it is low in volatility so as to be able to stay for a sufficient time to exhibit a skin condition improving effect, the active ingredient remains stable on the composition or skin, is easy to be formulated into cosmetics, and is safe for skin.

However, among the known components, components that satisfy all of the above characteristics are not common. For example, some skin condition-improving ingredients have excellent skin-improving activity even at low concentrations in in vitro experiments, but they are difficult to apply to real skin because of their ability to permeate through the skin. Other functional ingredients are poor in hydrophilicity and are difficult to formulate into cosmetics, or when they are exposed to heat, light, or oxygen, the active ingredient may be degraded or transformed into other compounds and the effect may already be lost before application to the skin.

The present inventors have made intensive studies to overcome the problems of the prior arts as a result, and as a result, it has been found that a composition for external application for skin comprising kirenol promotes the production of hyaluronic acid in keratinocyte, and hyaluronic acid synthase, HAS) and increase the synthesis of hyaluronic acid in the body. In addition, it disappeared free radicals and found to have excellent antioxidative effect, thus completing the present invention.

The Kastrup play (Kirenol) used in the present invention are hair jindeukchal (Siegesbeckia glabrescens ) extract.

As can be seen in the following examples, the pyrethroids show remarkably excellent hyaluronic acid production promoting effect or antioxidative effect at a low concentration. Therefore, chironol can be used as an active ingredient of a skin external composition for skin moisturizing or antioxidant effect.

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

The term "skin moisturizing" of the present invention refers to maintaining the homeostasis of a living body by appropriately adjusting water loss (moisture evaporation) and the like. When hyaluronic acid or collagen in the skin decreases, moisture is lost, hyaluronic acid or collagen If it is increased, moisture may increase.

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

The present invention relates to < RTI ID = It is based on the fact that expression of hyaluronic acid protein can be increased in human keratinocytes. In the present invention, an experiment was conducted to examine the effect of stimulating the production of hyaluronic acid in the pyrenol, and as shown in Examples 2 to 4 of the present invention, the concentration of hyaluronic acid was increased when the pyrenol was treated, The expression of hyaluronic acid protein was increased by gene expression of the enzyme.

The term "hyaluronic acid" according to the present invention is a kind of glycosaminoglycans, and refers to a chain-like polymer polysaccharide substance in which glucuronic acid and N-acetylglucosamine residues are repeatedly connected.

The present invention also provides a composition for external application for skin for antioxidation, which comprises as an active ingredient, a quinolone. The composition for external application for skin for antioxidation is characterized by exhibiting an antioxidative effect by inhibiting radical formation. The composition for external application for skin for antioxidation can effectively remove active oxygen generated from outside the living body or generated in living body, and thus can be used for skin anti-aging or anticancer.

In the present invention, an experiment was conducted to examine the radical formation inhibitory effect of the pyrenol. As shown in Example 5 of the present invention, when the pyrenol was treated, it showed a higher radical scavenging activity than the ascorbic acid, Excellent.

The term "antioxidant" effect of the present invention inhibits the oxidation of cells with highly reactive free radicals or reactive oxygen species (ROS) according to the oxidative stress caused by intracellular metabolism or ultraviolet rays. And includes removal of free radicals or reactive oxygen species, thereby reducing damage to the cells.

The skin external application composition for skin moisturizing or antioxidation according to the present invention can 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 comprises 0.00001 to 10% by weight, preferably 0.0001 to 7% by weight, more preferably 0.001 to 5% by weight, and most preferably 0.01 to 3% by weight, %. When the concentration is less than 0.00001% by weight based on the total weight of the external composition for skin, it is difficult to obtain the effect. When the concentration exceeds 10% by weight, problems may arise in formulation stability.

The present inventors have found that chironol increases the protein expression of hyaluronic acid in the skin to promote its production and has an antioxidative effect, so that it can be effectively used as an active ingredient of 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 form that can be applied to a 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, an external ointment, a cream, a foam, a nutritional lotion, a soft lotion, a perfume, a pack, a soft water, a latex, a makeup base, an essence, Or a powder selected from the group consisting of sunflow oil, suspension, emulsion, paste, gel, lotion, powder, soap, surfactant-containing cleansing oil, powder foundation, emulsion foundation, wax foundation, patch, Can be used.

The composition for external application for skin containing kirenol according to the present invention may contain cosmetic or pharmacologically acceptable carriers, diluents, adjuvants, coloring agents, stabilizers, stabilizers and the like to facilitate the use and handling, , A flavoring agent, a surfactant, an oil, a humectant, an alcohol, a thickening agent, an antioxidant, a pH adjusting agent, or an ultraviolet screening agent.

The composition for external application for skin, which comprises the cholineol according to the present invention as an active ingredient, accelerates hyaluronic acid production and expression of a synthetic enzyme, thereby promoting skin moisturization and eliminating free radicals to exhibit an excellent antioxidative effect. It can be used as an active ingredient.

Hereinafter, embodiments of the present invention will be described in detail to facilitate understanding of the present invention. However, the embodiments according to the present invention can be modified into various other forms, and the scope of the present invention should not be construed as being limited to the following embodiments. Embodiments of the invention are provided to more fully describe the present invention to those skilled in the art.

Preparation of materials

Kirenol used in the following examples was purchased from Shaanxi Shipyard Biological Co., Ltd., China.

Example  One: Pyrenyl  Cytotoxicity Assessment

Cell viability of human keratinocytes (HaCaT) was measured by Cell Counting Kit-8 (Dojindo Lab, USA) for evaluation of cytotoxicity of chironol.

First, a uniform number of cells was plated on a 96-well plate, stabilized, and then treated with 0.5 or 1 ppm of choline to further culture for 24 hours. After that, CCK-8 was treated and the absorbance was measured at 450 nm for 2 hours. The relative cell viability of cells treated with the quinolone versus 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 Chirenol
0.5 ppm Chirenol
1 ppm Survival rate (%) 100 96 98 94

* Number of iterations n = 3

* Relative cell survival rate = (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 the treatment with chelinol. This means that Kireonol does not cause cytotoxicity.

Example  2: Enzyme-linked immunosorbent assay (ELISA) was used in human keratinocytes To Kirenol  Increased hyaluronic acid concentration by

Human keratinocytes were cultured in DMEM medium containing 10% fetal bovine serum at a concentration of 1 × 10 ⁵ cells / ml and 500 μl / well in a 24-well plate. After incubation for 18 hours, the serum was not contained Washed twice with non-DMEM medium, and added to the concentration of chironol (0.5 and 1 ppm). Negative controls were treated with the same amount of ethanol and treated with 100 ppm of glucosamine hydrochloride (Sigma-aldrich), a positive control known to promote hyaluronic acid production. After culturing for 24 hours, cell culture medium was recovered and the concentration of hyaluronic acid was measured using a Hyaluronan ELISA kit (R & D Systems).

sample No treatment
(Negative control) Glucosamine hydrochloride
100 ppm Chirenol
0.5 ppm Chirenol
1 ppm Hyaluronic acid
Relative concentration (%) 100 139 131 169

* Number of iterations n = 3

* The concentration of hyaluronic acid in the experimental group treated with the chelonol was calculated based on the hyaluronic acid concentration of the negative control group (100%).

As a result, chironol increased hyaluronic acid production in human keratinocytes in a concentration-dependent manner.

Example  3: From keratinocytes derived from human body To Kirenol  Of hyaluronic acid synthase (HAS) gene

Human keratinocytes were cultured in DMEM medium containing 10% fetal bovine serum at a density of 1 × 10 ⁶ cells / ml, and 3000 μl / well of a 6-well plate was cultured for 18 hours. Washed twice in DMEM medium, and treated with 0.5 ppm and 1 ppm of chelinol for 24 hours. Cells were collected, and total RNA was extracted with RNeasy mini kit (Qiagen), and then 1 μg of RNA was quantified and reverse transcribed using GeneAmp ^® RNA PCR kit (Applied Biosystems). The reverse transcription reaction was performed using a Mycycler ^® PCR instrument (Biorad).

After using the reverse transcription reaction solution 2 ㎕ and HAS-2, HAS-3, Taqman ® probe (Invitrogen, USA, HS02511055_S1, HS03929097_g1) of GAPDH (Glyceraldehyde-3- phosphate dehydrogenase) was performed quantitative real time PCR. Real time PCR was performed using a CFX96 Touch ^TM Real-Time PCR Detection System instrument (Biorad). Experimental results obtained by 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 test group treated with the quinolone was quantified by using the amount of mRNA expression of the negative control as a reference (100%).

sample No treatment
(Negative control) Glucosamine hydrochloride
100 ppm Chirenol
0.5 ppm Chirenol
1 ppm HAS-2
Expression level (%) 100 125 109 119 HAS-3
Expression level (%) 100 130 106 125

* Number of iterations n = 3

As a result, expression of hyaluronic acid synthase, particularly HAS-2 and HAS-3, was increased in keratin-treated keratinocytes.

Example  4: From human-derived fibroblasts To Kirenol  Increased hyaluronic acid concentration by

Human Dermal Fibroblast was cultured in DMEM medium containing 10% fetal bovine serum at 1 × 10 ⁵ cells / ml and 500 μl / well in a 24-well plate. After 18 hours of culture, Lt; RTI ID = 0.0 > DMEM < / RTI > Chironol was added by concentration (0.5 and 1 ppm). Negative controls were treated with the same amount of ethanol and treated with 100 ppm of glucosamine hydrochloride (Sigma-aldrich), a positive control known to promote hyaluronic acid production. After culturing for 24 hours, the cell culture medium was recovered and the concentration of hyaluronic acid was measured using a Hyaluronan ELISA kit (R & D Systems).

sample No treatment
(Negative control) Glucosamine hydrochloride
100 ppm Chirenol
0.5 ppm Chirenol
1 ppm Hyaluronic acid
Relative concentration (%) 100 167 189 206

* Number of iterations n = 3

From the results of the experiments shown in Table 4, the chorinol increased hyaluronic acid production in human fibroblasts.

Example  5: To Kirenol  Antioxidant Effect - Free Radical Scavenging Rate

Free radical scavenging activity was measured to confirm the antioxidative activity of chironol. The free radical scavenging activity was determined by the 1,1-diphenyl-2-picryl-hydrazyl (DPPH) method as follows (Blois, MS Nature 181, 1190, 1958). DPPH is a relatively stable free radical, exhibiting maximum absorption at 517 nm in the presence of radicals, and loses its absorbance when cleaved. DPPH was purchased from Sigma, and dissolved in methyl alcohol at a concentration of 0.15 mM. First, 100 [mu] L of each of the wells of a 96-well plate was loaded with chironol and ascorbic acid as a positive control. 100 μl of DPPH solution was added thereto, and the mixture was allowed to stand at room temperature for 30 minutes. Then, the absorbance at 517 nm was measured using a microplate reader (BioTek EL-340). At this time, methyl alcohol was used instead of the quinolone as a control group. The effect of free radical scavenging activity was determined with the absorbance measured for the experimental group and the control group, and the results are shown in Table 5 below.

Free radical scavenging rate (%) sample Free radical scavenging rate (%) Chirenol 56.2 Ascorbic acid 78.9 Methyl alcohol 8.4

As can be seen from the results in Table 5, it is known that chironol exhibits antioxidative effects because it shows high activity as compared with ascorbic acid, which is a known antioxidant.

Example  6: To Kirenol  Increased skin moisture by

In order to apply the skin moisturizing effect to the external skin preparation containing the quinolone to human skin, it was prepared according to the composition of the nutritional cream of Preparation Example 1 below. In a constant temperature and humidity chamber at 22 ° C and a relative humidity of 50% (2 mg / ㎠) was applied to the inside of the habock, and the moisture content of the skin was measured before application, one hour after application and two hours after application. In the control group, nutritional cream containing the same amount of ethanol instead of chelenol was prepared and applied in the same manner, and normal skin without any treatment was used as a non - treated group. The water conductivity was measured by using a Corneometer (Courage + Khazaka, GmbH, Germany). The results are shown in Table 6 below.

Units (corneometer units) Production Example 1
Nourishing cream Control group
Nourishing cream Untreated group Before application 53 55 57 After 1 hour of application 84 79 55 After 2 hours of application 72 65 54

* n = 20

As a result, the nutritional cream containing Kirenol increased the water content of human skin and showed excellent water holding ability even after 2 hours of application.

As a result, it has been confirmed that quinolone has an excellent skin moisture increasing effect and water retention ability and can be useful for skin moisturizing.

The production example of the composition according to the present invention will be described in detail based on the above-mentioned effect test results. However, it should be apparent to those skilled in the art that the following preparations are merely examples of the composition of the present invention, and that the composition of the present invention is not limited to the following formulations. Hereinafter, a prescription example of an external preparation for skin containing a quinolone is shown by a production example.

Manufacturing example  1: Manufacture of nutritional cream

Nutritional creams containing the chelnol as an active ingredient in the composition shown in Table 7 below were prepared by a conventional method used in the field of cosmetics.

Ingredients Content (% by weight) Chirenol 1.0 Beta-1,3-glucan 5.0 Wax 10.0 Polysorbate 60 1.5 Piggy 60 hardened castor oil 2.0 Sorbitan sesquioleate 0.5 Liquid paraffin 10.0 Squalane 5.0 Caprylic / capric triglyceride 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, it will be understood by those skilled in the art that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. In this regard, it should be understood that the above-described embodiments are to be considered in all respects as illustrative and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention without departing from the scope of the present invention as defined by the appended claims.

Claims (5)

A composition for external application for skin for moisturizing skin, comprising Kirenol as an active ingredient. The composition for external application for skin for skin moisturizing according to claim 1, wherein the composition promotes hyaluronic acid production. A composition for external application for skin for antioxidant comprising Kirenol as an active ingredient. 4. The composition for external application for skin according to any one of claims 1 to 3, wherein the composition comprises 0.001 to 10% by weight of choline in a total weight of the composition. 4. The composition according to any one of claims 1 to 3, wherein the formulation of the composition is selected from the group consisting of a solution, an ointment, a cream, a foam, a nutritional lotion, a flexible lotion, a perfume, a pack, The compositions of the present invention may be formulated as tablets, pills, powders, creams, bath salts, sunscreen creams, sun oils, suspensions, emulsions, pastes, gels, lotions, powders, soaps, surfactant- containing cleansers, oils, powder foundations, emulsion foundations, waxes, Wherein the composition is any one selected from the group consisting of: