KR20180087479A

KR20180087479A - Composition for skin barrier comprising extract of Kaki Calyx

Info

Publication number: KR20180087479A
Application number: KR1020170010174A
Authority: KR
Inventors: 김민지; 윤석균; 이경은; 남진주; 박지은; 노윤화; 강승현; 김연준
Original assignee: 코스맥스 주식회사
Priority date: 2017-01-23
Filing date: 2017-01-23
Publication date: 2018-08-02

Abstract

The present invention relates to a composition for strengthening skin barrier containing a persimmon stalk extract. According to the present invention, the composition for strengthening skin barrier containing the persimmon stalk extract increases expression of filaggrin and claudin-1 as a skin moisturizing factor. By inhibiting expression of thymic stromal lymphopoietin (TSLP), thymus and activation-regulated chemokine (TARC) and macrophage-derived chemokine (MDC) which are skin inflammation inducing factors, it is possible to strengthen skin barrier by preventing skin inflammation and reducing loss of moisture in skin.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for enhancing skin barrier containing a persimmon extract,

The present invention relates to a composition for strengthening skin barrier.

The skin performs various functions essential for the human body to survive. Barriers to maintain homeostasis in the body in response to environmental changes, sensory functions to perceive external changes, and temperature control functions are among the most representative skin functions. Among the various functions of the skin, in particular, the barrier function of the skin is mainly caused by the stratum corneum which is located at the outermost part of the skin. The stratum corneum has been reported to affect not only the barrier function but also the function, role and structure of the inner living cell layer, ie, the epidermis or dermis, and its importance is continuously increasing. This stratum corneum is composed of dead keratinocyte and intercellular lipids, and plays a key role as a skin barrier that protects skin from external stimuli and prevents water from evaporating inside. In addition, the keratinocyte of the stratum corneum produces a skin barrier through differentiation and keratinization processes.

There are various factors that cause aging of human skin, but ultraviolet rays cause skin wrinkles, decrease of elasticity, pigmentation and skin barrier damage due to skin barrier damage. When skin moisture is reduced due to ultraviolet rays, the skin becomes dry due to loss of elasticity of the stratum corneum, which eventually prevents the skin from functioning as a barrier. Therefore, in order to strengthen the skin barrier, it is very important to maintain the moisture of the skin.

There are two types of pores associated with the water homeostasis present in the epidermis: Aquaporins (AQP) and Tight Junction (TJ). Among them, Tight Junction and Tight Junction-proteins serve as normal barriers to prevent TEWL in healthy skin and function as a structural system by up-regulating when the skin is damaged or wounded. Specifically, tight junctions (TJs) are located in the intercellular space of the epidermal granule cell layer, located at the top of the epidermis, and play a beneficial role in protecting the internal organs from the external environment and maintaining the homeostasis of the human body do. Such TJ constructs include TJ-associated proteins such as occludin (OC), claudin, and junctional adhesion molecules. They are mediated by plaque proteins (zona occludens-1, cingulin) present in the cytoplasm and function as biological barriers to control the adhesion of solutes and water transfer through the periplasmic space function.

Filagrin is a precursor protein of the natural moisturizing factor (NMF) responsible for skin moisturization. It is known that the promoting activity of filla green plays an important role in moisturizing action (J. Invest. Dermatol. 54, 24-31, 1970).

Meanwhile, various inflammatory dermatitis is caused by IgE-related immunity. In this case, there are many reports that delayed immune response due to T-cell abnormality is involved. Particularly, at the site where atopic dermatitis occurs, infiltration of immune-related cells such as macrophages, Th lymphocytes, and mast cells is greatly increased. In atopic dermatitis patients with atopic dermatitis, the concentration of IgE in the blood is high because it increases the number of Th2 cells and stimulates the secretion of IgE through B lymphocyte stimulation of Th2 cytokines such as IL-4 and 13 secreted by these cells . IL-4 and IL-13 play an important role in early atopic dermatitis (Donald Y. M. Leung et al., J Clin Invest. 2004, 113, 651-657).

Thymic stromal lymphopoietin (TSLP) is a top-level protein that causes immune imbalance by abnormalities in various biological signal transduction. It is produced by keratinocytes, fibroblasts or mast cells that constitute the skin when exposed to external antigens or microorganisms. Lt; / RTI > The resulting TSLP causes the immune system to be defeated by Th2 through a variety of immune mechanisms. In particular, dendritic cells (DC) are activated to produce th2-attracting chemokines such as thymus and activation-regulated chemokine (TARC) and macrophage-derived chemokine (MDC). Recent reports indicate that TSLP plays an important role in a variety of inflammatory dermatitis, especially atopic dermatitis. In addition, serum concentrations of MDC and TARC have been reported to be closely correlated with a variety of inflammatory dermatitis, particularly Th2 cell-associated skin diseases such as atopic dermatitis (Y. Shimada et al., J. Dermatol. , 2004).

For the treatment of atopic dermatitis, moisturizer should be applied regularly, and when severe, steroid ointment or antihistamine should be prescribed. This may temporarily alleviate symptoms, but steroid hormones may cause skin atrophy or enlargement of the capillaries when used for a long period of time. Rather, it may cause side effects such as weakening of skin barrier function and osteoporosis.

Accordingly, there is a demand for the development of a natural remedy that can enhance the skin barrier function by moisturizing the skin and alleviating inflammation of the skin.

The nipple (body,

) Is a deciduous shrub belonging to the family Ebenaceae (枾樹 tree) Diospyros kaki THUNB. [KHPCP] refers to the calyx of the fruit calyx. The calyx is thin calyx and narrow calyx with a diameter of 15 ~ 25 ㎜ and a thickness of 1 ~ 4 ㎜. The lobe is ovate or triangular and wide ovate, mostly dried upwards, and the center of the lamella is thick. The outer surface is grayish brown or reddish brown, often with four ridges, some with residual disease, or with a recessed spot. When you look at the lamina on the inner side of the loupe, you will see brown short hairs in the middle and fruit in the center. Ursolic acid, oleic acid, betulic acid, and tannic acid are considered to be effective for cough, asthma and chronic bronchitis as well as for hiccups In addition, However, to date, there has been no known use for enhancing the skin barrier of the tap root extract. Accordingly, the inventors of the present invention discovered that the ginseng extract had such an effect while developing a natural skin barrier enhancer which is safe to human body.

As a result, the present inventors have made efforts to overcome the problems of the prior arts. As a result, the present inventors have found that, in the case of a composition for enhancing skin barrier containing the extract of persimmon extracts, the skin moisturizing factors, such as claudin- Inhibiting the expression of TSLP, TARC, and MDC as the factors, thereby reducing the skin moisture loss and preventing skin irritation. Thus, the present inventors have completed the present invention.

KR 10-2009-0098732 A KR 10-2005-0063164 A

Accordingly, it is a main object of the present invention to provide a composition for enhancing skin barrier containing a persimmon extract capable of reducing skin moisture loss and preventing skin irritation, thereby enhancing skin barrier.

According to one aspect of the present invention, there is provided a composition for skin barrier enhancement comprising as an active ingredient a gum nut extract.

The term " strengthening the skin barrier " of the present invention means strengthening the barrier of the stratum corneum located on the outer surface of the skin. In addition to being easily damaged by the external environment, the stratum corneum, which is the primary barrier of the skin, can not function properly or is damaged by inflammatory dermatitis such as atopy or dry skin. In order to moisturize dry skin, it is usually possible to solve temporary problems through a prescription of moisturizing agent, but it is difficult to solve fundamental problems, and inflammatory dermatitis such as atopic dermatitis may prescribe steroid to temporarily relieve inflammation. It is also a temporary measure, but a prescription like steroids rather weakens skin barrier function. Accordingly, the inventors of the present invention confirmed that the gut peel extract of the present invention can solve the fundamental problem and enhance the skin barrier, thus completing the present invention.

In the present invention, the extract may be extracted with any solvent conventionally used for extracting natural plants, preferably water, a lower alcohol having 1 to 4 carbon atoms, butylene glycol, propylene glycol, glycerin or the like , And more preferably, it can be extracted with a polar solvent of a lower alcohol such as ethanol.

The chewing gum extract of the present invention can be obtained by collecting the gut buds, drying and pulverizing them, and then heating the mixture at 60 to 100 ° C for about 0.1 to 12 hours with a polar solvent having a volume of about 1 to 20 times the weight of the sample, , Preferably for 1 to 6 hours, and then filtering and concentrating the obtained extract.

In the present invention, the extract may be contained in an amount of 0.001 to 30% by weight, preferably 0.1 to 10% by weight, based on the total weight of the composition

In the present invention, the extract is characterized by increasing the expression of claudin-1 and / or filaggrin of keratinocytes.

According to the experimental example of the present invention, the expression of claudin-1 and filaggrin, which are skin moisturizing factors, was confirmed in order to confirm whether moisturizing power can be given by reducing moisture loss of skin. As a result, -1 and pilar green were increased. From these results, it is suggested that the gut peel extract of the present invention increases the expression of a skin moisturizing factor and ultimately gives moisturizing power to the skin, thereby enhancing the skin barrier (Experimental Example 3, Figs. 5 and 6 ).

In the present invention, the extract is characterized by inhibiting expression of thymic stromal lymphoietin (TSLP), thymus and activation-regulated chemokine (TARC), and macrophage-derived chemokine (MDC).

According to the experimental example of the present invention, the extracts of Poly I: C and IL-4, which induce TSLP, TARC, and MDC were significantly inhibited by the concentration of persimmon extracts. From these results, it is suggested that the composition of the present invention inhibits the expression of TSLP, TARC and MDC, thereby preventing or improving dermatitis resulting from immune loss, thereby enhancing skin barrier (Experimental Examples 1 to 2 To 4).

In the present invention, the skin barrier strengthening is characterized by reducing water loss of the skin and preventing inflammation of the skin.

In the present invention, the composition is characterized by being a composition for external application for skin or a cosmetic composition.

In the present invention, the composition may be in the form of a lotion, a cream, a cream, a cream, The composition is characterized in that it has one formulation selected from the group consisting of nutritional essence, pack, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, lotion, ointment, gel, cream, patch and spray.

The ingredients contained in the external preparation for skin or cosmetic composition of the present invention may contain, as an active ingredient, the components commonly used in external preparations for skin or cosmetic compositions, such as stabilizers, solubilizers, vitamins, pigments and And customary adjuvants and carriers such as perfumes.

INDUSTRIAL APPLICABILITY As described above, the chewing gum extract of the present invention can enhance the skin barrier by reducing moisture loss of the skin and inducing inflammation of the skin, and it is also possible to provide a skin external application Composition or cosmetic composition may be prepared.

FIG. 1 is a graph showing the results of Experimental Example 1-2 in which the effect on the expression of TSLP gene in the treatment of persimmon extract was measured.
FIG. 2 shows the results of Experimental Example 1-3 in which the effect on the expression of TSLP protein in the treatment of persimmon extract was measured.
FIG. 3 is a graph showing the results of Experimental Example 2 in which the effect on the TARC gene expression was examined in the case of treating the persimmon extract. FIG.
4 is a graph showing the results of Experimental Example 2 in which the effect on the expression of MDC gene in the treatment of persimmon extract is measured.
5 is a graph showing the results of Experimental Example 3 in which the effect of filamentous fungus extract treatment on the expression of filaggrin gene was measured.
FIG. 6 is a graph showing the results of Experimental Example 3 in which the effect on the expression of claudin-1 gene in the treatment of persimmon extract was measured.

Hereinafter, the present invention will be described in more detail with reference to Examples. These embodiments are only for illustrating the present invention, and thus the scope of the present invention is not construed as being limited by these embodiments.

Example One: A spigot Isolation and Identification of Extracts

Ganjangjip was purchased from Kyungdong market (Seoul, Korea) and dried. The dried persimmon powder was pulverized, and then 100 ml of 70% ethanol was added per 10 g of each, and then aged at room temperature for 1 day. Then, the extracts were sequentially filtered and concentrated under reduced pressure to prepare an extract.

Experimental Example 1: TSLP Inhibition Test

1-1. Human skin keratinocyte cell line HaCaT cell culture

TSLP (thymic stromal lymphopoietin) is classified as a top-level cytokine that initiates type 2 helper T cell-mediated immunity deficits that are recognized as the cause of atopic dermatitis. In order to confirm the reduction effect of Example 1 on TSLP (thymic stromal lymphopoietin) expression of human keratinocyte HaCaT, gene expression level of TSLP was evaluated.

The keratinocyte HaCaT, a constitutive cell line of human skin, was cultured in a DMEM culture medium containing 10% fetal bovine serum (FBS) and antibiotics. The cells were cultured in a 75 ° C flask and a 6-well plate at 37 ° C with 5% CO ₂ . The culture medium was changed every 3 to 4 days. When the cells were overproduced, they were subcultured. Cells were washed with phosphate-buffered saline solution (PBS) for 24 hours after culturing, and treated with Poly I: C, IL-4 in DMEM medium without FBS in each well, 1 was diluted to a final concentration of 1, 10 ppm, and added to each well. After 4 hours, cDNA was synthesized and PCR was performed to evaluate gene expression level or western blotting after 24 hours.

1-2. Test for effect of TSLP expression using real-time PCR

The synthesized cDNA was subjected to real-time PCR (PCR) using a master mix and a SYBR green master mix as a target template and a cyanine dye to finally determine the gene expression level of TSLP . Expression of the gene was compared with a relative value by calibrated quantification of the internal control gene β-actin by the Ct (threshold cycle) method. RT-PCR was performed using TSLP, TARC, MDC, Filaggrin, Claudin 1, and β-actin primers and cDNA with Cyber Green (SYBR Green supermix, Applied Biosystems, USA) Followed by polymerase chain reaction at 94 ° C for 5 minutes, polymerization at 95 ° C for 30 seconds, 55 ° C for 30 seconds, and 72 ° C for 30 seconds. The primer sequences are shown in Table 1 below.

Name of the primer order TSLP Forward 5'-GCTATCTGGTGCCCAGGCTAT-3 ' TSLP Reverse 5'-CGACGCCACAATCCTTGTAAT-3 ' TARC Forward 5'-CTTCTCTGCAGCACATCC-3 ' TARC Reverse 5'-AAGACCTCTCAAGGCTTTG-3 ' MDC Forward 5'- GCATGGCTCGCCTACAGACT-3 ' MDC Reverse 5'-GCAGGGAGGGAGGCAGAGGA-3 ' Filaggrin Forward 5'-AGTGCACTCAGGGGGCTCACA-3 ' Filaggrin Reverse 5'-CCGGCTTGGCCGTAATGTGT-3 ' Claudin1 Forward 5'-GCTCTAGAATTCCGAGCGAGTCATGGCCAACGC-3 ' Claudin1 Forward 5'-GCTCTAGAATTCTCACACGTAGTCTTTCCCGCT-3 ' β-actin Forward 5'-GGCCATCTCTTGCTCGAAGT-3 ' β-actin Reverse 5'-GACACCTTCAACACCCCAGC-3 '

As a result, as shown in FIG. 1, it was confirmed that TSLP overexpressed by poly I: C and IL-4 was significantly inhibited by treatment with 1, 10 ppm of tap root extract.

1-3. Test for effect of TSLP expression using Western blot

Cells were lysed with lysis buffer containing 20 mM Tris, 100 mM NaCl, 5 mM MgCl ₂ , 1% NP40, 0.5% sodium deoxychlolate, 10 mM glycerophosphate, 0.1 mM orthovanadate and protease inhibitor for 10 minutes and incubated at 13000 rpm for 10 minutes The proteins were separated by centrifugation. Separated proteins were quantified using the Bradford assay and the same amount of protein per sample was separated by electrophoresis on 10-12% SDS-PAGE and then transferred to the PVDF membrane. Membrane was blocked with 5% skim milk and blotted with TSLP and β-actin antibody and detected with ECL kit.

As a result, as shown in FIG. 2, it was confirmed that the TSLP protein overexpressed by poly I: C and IL-4 was significantly inhibited by 1, 10 ppm treatment of persimmon extract.

Experimental Example 2: Test for effect of expression of TARC and MDC using real-time PCR

The degree of inhibition of TARC and MDC mRNA expression by the tap extract was measured by the same method as in Experimental Example 1-2.

As a result, as shown in FIGS. 3 and 4, it was confirmed that the TARC and MDC genes overexpressed by poly I: C and IL-4 were significantly inhibited by treatment with 1, 10 ppm of tap root extract.

Experimental Example 3: Test for effect of filaggrin and claudin-1 expression using real-time PCR

The degree of filaggrin and claudin-1 mRNA expression by the tap extract was measured by the same method as in Experimental Example 1-2.

As a result, it was confirmed that filaggrin and claudin-1 genes reduced by poly I: C and IL-4 were significantly increased by treatment with 10 ppm of persimmon extract, as shown in FIGS.

Formulation Example 1: Preparation of a cream preparation

To prepare a cream preparation containing a gentle tap extract, it was prepared by a conventional method according to composition components and composition ratios as shown in Table 2.

ingredient Content (% by weight) Persimmon extract One Glyceryl stearate / phage-100 stearate 2.0 Polysorbate 60 0.5 Sorbitan stearate 0.5 Phitosqualan 5.0 Jojoba oil 3.0 Caprylic / capric triglyceride 10.0 Aloe butter 3.0 Wax 1.0 Stearyl alcohol 0.5 Beeryl alcohol 0.5 Stearic acid 0.5 glycerin 7.0 Butylene glycol 5.0 Carboxyvinyl polymer 0.2 Arginine 0.3 Dimethicone 0.5 antiseptic a very small amount Spices a very small amount Purified water Balance

Formulation Example 2: Preparation of lotion agent

Lotion agents containing the ingredients obtained in Example 1 were prepared according to the conventional methods according to the compositional components and composition ratios shown in Table 3 below.

ingredient Content (% by weight) Persimmon extract One glycerin 3.0 Butylene glycol 2.0 C14-22 Alcohol / C12-20 alkyl glucoside 2.0 Glycerin monostearate 0.5 Caprylic / capric triglyceride 10.0 Liquid paraffin 5.0 Carboxyvinyl polymer 0.2 Triethanolamine 0.2 antiseptic a very small amount Spices a very small amount Purified water Balance

Formulation Example 3: Preparation of Ointment Agent

Ointment agents containing the ingredients obtained in Example 1 were prepared in a conventional manner according to the compositional components and composition ratios shown in Table 4 below.

ingredient Content (% by weight) Persimmon extract One Wax 10.0 Polysorbate bit 60 1.5 Sorbitan sesquioleate 0.8 Liquid paraffin 40.0 Squalane 5.0 Caprylic / capric triglyceride 4.0 glycerin 5.0 Butylene glycol 3.0 Carboxyvinyl polymer 0.2 Triethanolamine 0.2 antiseptic a very small amount Spices a very small amount Purified water Balance

Claims

A composition for enhancing skin barrier comprising an extract of Kaki Calyx as an active ingredient.

The skin barrier enhancer according to claim 1, wherein the extract is extracted with water, at least one solvent selected from the group consisting of lower alcohols having 1 to 4 carbon atoms, butylene glycol, propylene glycol, glycerin, / RTI >

The composition for enhancing skin barrier according to claim 1, wherein the extract comprises 0.1 to 10% by weight based on the total weight of the composition.

The composition for enhancing skin barrier according to claim 1, wherein the extract increases the expression of claudin-1 and filaggrin of keratinocytes.

The composition for enhancing skin barrier according to claim 1, wherein the extract inhibits the expression of thymic stromal lymphoietin (TSLP), thymus and activation-regulated chemokine (TARC), and macrophage-derived chemokine (MDC).

The skin barrier strengthening composition according to claim 1, wherein the skin barrier strengthening reduces moisture loss of the skin and prevents inflammation of the skin.

The skin barrier strengthening composition according to claim 1, wherein the composition is a composition for external application for skin or cosmetic composition.

The composition according to claim 1, wherein the composition is at least one selected from the group consisting of a skin lotion, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, a nutrition cream, a moisturizer cream, a hand cream, an ointment, Wherein the composition has one formulation selected from the group consisting of soaps, cleansing foams, cleansing lotions, cleansing creams, body lotions, body cleansers, lotions, ointments, gels, creams, patches and sprays.