KR101860352B1 - A cosmetic composition for treating demodex infestations comprising extract of artemisia apiaceae - Google Patents

Abstract

Provided is a cosmetic composition containing an Artemisiae Annuae Herba extract, a ginkgo leaf extract, and a fig extract as an active ingredient. More specifically, the purpose of the present invention is to provide a cosmetic composition with excellent biosafety as well as functionality.

Description

TECHNICAL FIELD [0001] The present invention relates to a cosmetic composition for removing folliculitis, which contains an extract of Cheongho Composition as an active ingredient,

The present invention relates to a cosmetic composition comprising a natural plant extract as an active ingredient.

Skin is the primary barrier of the human body. It protects the internal organs of the body from external stimuli such as changes in temperature and humidity, ultraviolet rays, and pollutants, and plays an important role in the maintenance of body homeostasis such as body temperature control.

Microcystic mite is an ex vivo parasite that is often observed under a microscope that infects the fur branching unit of the skin. Of the various species reported, at least Demodex folliculorum and Demodex brevis are found on the outside of the human body.

Two kinds of folliculitis are common, preferentially to the same skin area of the face, cheeks, forehead, nose and outer ear canal, where active sebaceous secretion is favorable for environment and reproduction. The adult folliculitis mite has a life cycle of 14 to 18 days and eventually dies, and their multiplication depends on successful mating by adult mite.

Mating occurs at the follicular opening (near the skin surface). Thereafter, the pregnant females migrate to the sebaceous glands to lay eggs and become larvae and first nymphs. The first nymph goes to the opening of the hair follicle to become the second nymph, and goes out into the skin and enters the hair follicle again to become an adult.

Thus, if the adult epididymis can successfully mate and produce the next generation in the life cycle process, the degree of infection in the host will increase.

Fungicide infections do not appear in healthy children, but they increase in an age-dependent manner and are found in nearly 100% of middle-aged skin unless an abortion is taken. The incidence of fungal infection is higher in unhealthy skin than in normal skin. Excessive growth of these mites is associated with skin disease and hair loss.

The fungus is prevalent in limb mammals, especially livestock animals, specifically dogs, and can cause hair follicle infection. The hair follicle mite enters hair follicles and sebaceous glands of animals, often resulting in severe dermatitis, infection and discomfort.

On the other hand, the fungus is a mite that parasitizes in the hair follicles and sebaceous glands of the skin. It grows from one to several pores in a few pores.

The soap or cleanser used in cleansing will remain on the skin after cleansing, and the remaining fat in the skin will cause the growth of the hair follicle. The hair follicle can start hair loss and dry skin can turn into oily skin. Accordingly, there is a need to develop a composition that is free of side effects in the development of soaps, detergents, shampoos, and cosmetics, and that is harmless to the human body and that is effective in removing folliculitis.

In this regard, the cosmetics industry is developing a number of products using natural products to reduce skin irritation caused by various chemicals. In addition to low adverse effects on skin, natural materials have recently been increasingly developed as cosmetic raw materials, as consumers have become more receptive to cosmetics using natural materials. However, the cosmetic product containing the extract derived from natural origin obtained by a conventional method is not sufficiently functionalized, and there is a problem that the skin improving activity is not maintained constantly.

Accordingly, there is a continuing need for the development of cosmetic products derived from natural products superior in biostability and skin improvement effect with excellent functionality.

Disclosure of Invention Technical Problem [8] The present invention has been made to solve the above-mentioned problems of the prior art, and an object of the present invention is to provide a cosmetic composition having not only functionality but also biostability.

According to one aspect of the present invention, And ginkgo biloba extract or fig extract as an active ingredient.

In one embodiment, the composition may include 10 to 30 parts by weight of the blueberry extract, 10 to 30 parts by weight of the ginkgo leaf extract, and 10 to 30 parts by weight of the fig extract.

In one embodiment, the extract may be an extract obtained by fermentation by inoculating the Lactobacillus genus (Lactobacillus sp.) Strains.

In one embodiment, the extract is selected from the group consisting of water, anhydrous or lower alcohols having 1 to 4 carbon atoms, acetone, petroleum ether, ethyl acetate, butyl acetate, trichloromethane, dichloromethane, chloroform, Glycol, < / RTI >

In one embodiment, the composition may be for skin whitening, skin moisturizing or wrinkle improvement.

In one embodiment, the composition may be for removing folliculitis.

In one embodiment, the composition is comprised of a softener, a nutritional lotion, a water cream, a nutritional cream, a massage cream, an essence, an ampoule, a gel, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, ≪ / RTI >

According to the present invention, the cosmetic composition of the present invention contains not only the skin improving effect but also the human body safety because the natural complex extract contains the active ingredient.

It should be understood that the effects of the present invention are not limited to the effects described above, but include all effects that can be deduced from the description of the invention or the composition of the invention set forth in the claims.

As used herein, the terminology used herein is intended to encompass all commonly used generic terms that may be considered while considering the functionality of the present invention, but this may vary depending upon the intent or circumstance of the skilled artisan, the emergence of new technology, and the like. Also, in certain cases, there may be a term selected arbitrarily by the applicant, in which case the meaning thereof will be described in detail in the description of the corresponding invention. Therefore, the term used in the present invention should be defined based on the meaning of the term, not on the name of a simple term, but on the entire contents of the present invention.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Terms such as those defined in commonly used dictionaries are to be interpreted as having a meaning consistent with the contextual meaning of the related art and are to be interpreted as either ideal or overly formal in the sense of the present application Do not.

The numerical range includes numerical values defined in the above range. All numerical limitations of all the maximum numerical values given throughout this specification include all lower numerical limitations as the lower numerical limitations are explicitly stated. All the minimum numerical limitations given throughout this specification include all higher numerical limitations as the higher numerical limitations are explicitly stated. All numerical limitations given throughout this specification will include any better numerical range within a broader numerical range, as narrower numerical limitations are explicitly stated.

Hereinafter, embodiments of the present invention will be described in detail, but it should be apparent that the present invention is not limited by the following examples.

One aspect of the present invention relates to a composition comprising: And ginkgo biloba extract or fig extract as an active ingredient.

The cosmetic composition includes a natural plant extract as an active ingredient, so that the skin improving effect and the biosafety are excellent, and the skin irritation can be minimized.

The cosmetic composition is excellent in skin wrinkle improvement and skin moisturizing effect as well as skin peeling effect by the interaction of various active ingredients contained in the natural complex extract. In particular, the active ingredient of the above-mentioned Cheongho extract, Ginkgo biloba extract, and fig extract can effectively inhibit the growth of skin folliculitis by acting on the skin at the same time.

The above " Artemisiae Annuae Herba ) "is a species of Artemisiaannua Linne or Artemisia apiacea Hance ). It is also used for the treatment of fever and cold, schizophrenia, pediatric competition, dyspepsia and dysentery in one room. Also, it is known as a source of anti-malaria agent artemisinin.

The leaves of the " Ginkgo biloba L " contain various components. The ginkgo leaf contains flavonoids, biflavonoids, proanthocyanidins, terpenolactone, and the like, and bioflavonoids such as quercetin, kaempferol, and Isohamnetin, It is said that the most ripe green leaves are abundant when collected in September-October. It is known that the Ginkgo biloba extract inhibits the abnormal expression of melanocytes by inhibiting the expression of the stained blood vessels, which are found to be highly toxic to the spots.

The above " Ficuscarica L. " is a subtropical deciduous broad-leaved broad-leaved tree, belonging to the genus Phalaenopsis . More than 600 varieties are distributed worldwide, and it is known as the first fruit that humans have used for a long time. These figs have long been recorded in the Bible and Dongwoo-bombo as private medical uses and are known to be good for blood pressure strengthening, dryness, nourishment, constipation, hepatitis, cancer, and women's vitality recovery. The general efficacy of these figs is that they act as gastrointestinal drugs, diarrhea, treatment of hemorrhoids, papillary throat, inflammation, phlegm action, and digestion.

As used herein, the term " comprising as an active ingredient " means an effective amount that can exhibit a skin improving effect, for example, a degree of collagen synthesis associated with a wrinkle reducing effect, an elasticity improving or skin whitening effect, can do.

The above-mentioned "extract" refers to a solvent in which an effective ingredient contained in an extraction raw material has been transferred by contacting a solvent and an extraction raw material under specific conditions. If a substance is separated from a natural product, the extraction method, You can include them all regardless. For example, it may include extracts of components dissolved in a solvent from natural products by using water or an organic solvent, and specific components of natural products, such as oils obtained by extracting only specific components such as oil. The extract can be obtained by drying and pulverizing the raw material for extraction, or various extraction methods known in the art such as a hot water extraction method and an ethanol extraction method.

Wherein the extract is selected from the group consisting of water, an anhydrous or lower alcohol having 1 to 4 carbon atoms, acetone, petroleum ether, ethyl acetate, butyl acetate, trichloromethane, dichloromethane, chloroform, hexane and 1,3-butylene glycol (V / v) ethanol, in particular at a concentration of between 60 and 80%.

The extraction ratio of the active ingredient contained in the raw material may be different depending on the polarity of the solvent. Since the ethanol is excellent in selectivity for extracting a physiologically active substance from a natural raw material, the ethanol extract can realize an optimal skin improving effect .

Since the water and ethanol have different polarities and the effective components to be extracted depend on the polarity, the concentration of ethanol can be appropriately controlled so that an optimal skin improving effect can be realized. At this time, if the concentration of ethanol is less than 60%, the effective ingredient showing skin improving effect may not be extracted sufficiently, and if it is more than 80%, an adequate yield may not be realized.

The extract is washed with water, dried and pulverized, and then extracted with a solvent having a volume of 8 to 12 times the weight of the raw material for about 1 to 24 hours by a conventional method such as circulation extraction, pressure extraction or ultrasonic extraction And filtering. In addition, the extract can be obtained in powder form by an additional process such as vacuum distillation or freeze-drying.

The extract may also contain an extract which has been subjected to a conventional purification process. For example, the extract may be subjected to various purification methods such as separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity) Lt; RTI ID = 0.0 > fractions. ≪ / RTI >

The composition may include 10 to 30 parts by weight of the blueberry extract, 10 to 30 parts by weight of the ginkgo leaf extract, and 10 to 30 parts by weight of the fig extract.

When the variation of the mixing ratio of the extract is large, the skin improvement effect derived from the active ingredient of each extract may not be properly implemented, and thus the mixing ratio can be appropriately controlled in consideration of the working environment and the quality of the final product. The extract may be provided with an optimal skin improving effect derived from the active ingredient in the above weight range.

The extract may be an extract obtained by fermentation by inoculating the Lactobacillus genus (Lactobacillus sp.) Strains.

The term " fermentation " refers to a process in which a microorganism decomposes an organic substance using an enzyme contained therein. It is known that a raw material that has undergone fermentation by the microorganisms has an effect of at least 2 times to a maximum of several tens of times greater than that before fermentation. Fermented metabolites contain various amino acids, organic acids and antioxidants that are good for the skin, promoting skin metabolism and making skin texture resilient and smooth. In addition, it has been reported that as the fermentation process proceeds, the particles become smaller and the toxicity such as heavy metals is decomposed and the absorption rate is improved, and the skin troubles and allergic side effects are alleviated.

The "fermentation extract" is obtained by drying the extract naturally or by using an apparatus, specifically a rotary vacuum concentrator and a freeze drier, diluting the extract to a predetermined concentration in a solvent, preferably purified water, That is, in Lactobacillus (Lactobacillus sp.) may be prepared by inoculation of strains and fermentation.

The Lactobacillus genus strains Lactobacillus pento suspension (Lactobacillus pentosus), Lactobacillus brevis (Lactobacillus brevis), Lactobacillus Planta room (Lactobacillus plantarum), Lactobacillus casei (Lactobacillus casei) or Lactobacillus know also required Ruth (Lactobacillus acidophilus , but are not limited thereto.

The composition may be for skin whitening, skin moisturizing or wrinkle improvement, and may be for removing folliculitis. Due to the various active ingredients contained in the plant extract, the composition has excellent skin improving effect and can effectively inhibit the growth of the hair follicle.

The " wrinkles " refers to the scarring caused by degeneration of the skin, and may be caused by a cause of genes, a reduction of collagen and elastin present in the skin dermis, and an external environment. The " wrinkle improvement " refers to a phenomenon that inhibits or inhibits the generation of wrinkles on the skin, or alleviates the already generated wrinkles.

The term " skin moisturizing " means any action that maintains the homeostasis of the skin tissue by appropriately controlling moisture loss (moisture evaporation) of the skin and the like. The skin moisturizing effect may be accompanied by an additional skin improving effect in various aspects such as an exfoliation improving effect and a skin irritation reducing effect.

The "skin whitening" mentioned above is intended to mitigate the formation of various biomolecules such as melanin, redox hemoglobin, carotene and melanoid affecting the skin color, thereby alleviating skin pigmentation phenomena such as dullness, freckles and stains, It means an effect of alleviating redness and improving skin brightness and uniformity.

The composition may be at least one selected from the group consisting of softening agents, nutritional lotion, water cream, nutritional cream, massage cream, essence, ampoule, gel, eye cream, cleansing cream, cleansing foam, cleansing water, .

The cosmetic composition may be formulated by a conventional method. The contents disclosed in Remington ' s Pharmaceutical Sciences, Mack Publishing Company, Easton PA in the formulation of the external preparation for skin can be referred to, and in the formulation of the cosmetic composition, International cosmetic ingredient dictionary, 6th ed (The cosmetic, Toiletry and Fragrance Association , Inc., Washington, 1995).

Specifically, the cosmetic composition may be prepared into a general emulsified formulation and a solubilized formulation. For example, lotion such as soft lotion or nutrition lotion; Lotion such as facial lotion, body lotion; Nourishing cream, moisture cream, cream such as eye cream; essence; Makeup ointment; spray; Gel; pack; Sunscreen; Makeup base; A foundation such as a liquid type, a solid type or a spray type; powder; Make-up removers such as cleansing creams, cleansing lotions and cleansing oils; Or a cleansing agent such as a cleansing foam, a soap, a body wash, and the like, but is not limited thereto. The external preparation for skin may be formulated into ointments, patches, gels, creams or sprays, but is not limited thereto.

The cosmetic composition of the present invention may be appropriately blended with other ingredients within the range of not compromising the object of the present invention depending on the kind of the formulation or the purpose of use, in addition to the essential ingredients in each formulation.

The cosmetic composition may contain a conventional acceptable carrier and may appropriately contain, for example, oil, water, a surfactant, a moisturizer, a lower alcohol, a thickener, a chelating agent, a colorant, no.

The acceptable carrier may vary according to the formulation. For example, when formulated into ointments, pastes, creams or gels, there may be used an animal oil, a vegetable oil, a wax, a paraffin, a starch, a tracer, a cellulose derivative, polyethylene glycol, silicon, bentonite, silica, talc, zinc oxide, Mixture can be used

When the cosmetic composition is formulated into a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder or a mixture thereof may be used as a carrier component. In the case of a spray, Propellants such as carbon, propane, butane or dimethyl ether.

When the cosmetic composition is formulated into a solution or emulsion, a solvent, a solubilizing agent or an emulsifying agent may be used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl benzoate, 1,3-butyl glycol oil may be used, and in particular, fatty acid esters of cottonseed oil, peanut oil, corn oil, olive oil, castor oil and sesame oil, glycerol aliphatic ester, polyethylene glycol or sorbitan may be used.

When formulated into a suspension, the cosmetic composition may contain, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the cosmetic composition is formulated into soap, it is preferable to use, as a carrier component, an alkali metal salt of a fatty acid, a fatty acid hemiester salt, a fatty acid protein hydrolizate, isethionate, a lanolin derivative, an aliphatic alcohol, a vegetable oil, Can be used.

The cosmetic composition may be a cosmetic or dermatological composition which is generally used in the art depending on the quality or function of the final product, such as a lipid, an organic solvent, a solubilizing agent, a thickening agent, a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, Surfactants, water, ionic or nonionic emulsifiers, fillers, metal ion sequestrants, chelating agents, preservatives, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, And may further contain adjuvants conventionally used in the field of cosmetics or dermatology, such as any other ingredient.

However, the adjuvant and the mixing ratio thereof can be appropriately selected so as not to affect the preferable properties of the cosmetic composition according to the present invention.

The present invention will be further described with reference to the following examples, but it should be apparent that the present invention is not limited by the following examples.

Manufacturing example  1: Preparation of plant extracts

After washing with water, ginkgo, ginkgo leaf, and fig, they were thoroughly dried at room temperature and pulverized to obtain 100 g of pulverized products.

100 g of each pulverized product was immersed in a 10-fold volume of an aqueous ethanol solution (70% w / w) as a solvent at a temperature of 70 DEG C for 12 hours.

The extract was filtered through a 250 mesh filter and a 0.5 탆 filter, and then extracted three times in the same manner with respect to the remaining raw materials. The filtrate was concentrated under reduced pressure at 30 캜 and lyophilized to obtain a solid fraction.

Manufacturing example  2: Preparation of plant fermentation extract

Each of the extracts obtained in Preparation Example 1 was diluted to 5% by weight with purified water and then inoculated with a fermentation strain, Lactobacillus plantarum , and fermented for 12 hours.

The fermentation extract was filtered through a 500 mesh and 0.2 mu m filter to obtain fermented extracts, respectively.

Example  And Comparative Example

To examine the skin improving effect of the obtained extract and the fermented extract, examples and comparative examples were set by mixing samples as shown in Table 1 below.

The skin improvement effect was measured by mixing each raw material at a predetermined ratio as shown in Table 1 below. In Examples 4 to 6, fermented extracts of Preparation Example 2 were mixed.

[Content (parts by weight)] division Example Comparative Example One 2 3 4 5 6 One 2 3 4 Cheongho extract 50 50 50 - - - 100 - - - Ginkgo biloba extract 50 - 50 50 - - - 100 - 50 Fig extract - 50 50 - 50 - - - 100 50 Cheongho fermented extract - - - 50 50 50 - - - - Ginkgo leaf fermentation extract - - - - - 50 - - - - Fig fermented extract - - - - - 50 - - - -

Experimental Example  1: skin cell safety test

MTT assay experiments were performed on fibroblasts (HDF), melanocytes (B16F10), and keratinocytes (HaCaT) in order to verify the skin safety of the samples obtained in Preparation Examples 1 and 2.

Samples of Preparation Examples 1 and 2 were suspended in purified water at concentrations of 0, 1, 5, 10, 20, 50, and 100 μg / mL, and cell viability was measured.

Cytotoxicity was performed by modifying the method of Mosmann to measure cell viability using MTT {3- (4,5-dimethylthiazol-2-yl) -2-5-diphenyltetrazolium bromide} reagent.

(HDF), melanocyte-forming cells (B16F10), and keratinocytes (HaCaT) were dispensed in 96-well plates at a concentration of 1 × 10 ⁴ cells / well and cultured at 37 ° C. and 5% CO ₂ for 48 hours Respectively.

After the culture medium was removed and the sample was cultured for 48 hours in a concentration-treated medium, the medium was removed and repeatedly washed with PBS (phosphate buffered saline). MTT was dissolved in PBS at 5 mg / mL, and 50 L L was added and cultured at 37 째 C and 5% CO ₂ for 48 hours. 100 νL of DMSO (dimethyl sulfoxide) was added per well, and the mixture was stirred for 10 minutes and absorbance was measured at 540 nm.

No cytotoxicity was observed at all concentrations of each sample. The above results suggest that the extract of the above-mentioned example is not only harmless to human cells but also excellent in safety.

Experimental Example  2 : COL1A1  And Of MMP1  Expression test

Sample powders of Examples and Comparative Examples were suspended in purified water and diluted, and the effect of improving wrinkles was evaluated.

The expression of COL1A1, one of typical collagen expressed by human dermal fibroblasts, and MMP1, a collagenase, was measured.

In order to confirm the extent of expression of collagen, human dermal fibroblasts were treated with the extracts of the examples and comparative examples diluted to a concentration of 100 μg / mL and cultured for 24 hours.

The changes in gene expression were measured by qRT-PCR (quantitative real time PCR), which binds the fluorescent substance to the DNA product amplified by polymerase chain reaction (PCR) and continuously detects the fluorescent substance.

To quantify the changes of COL1A1 and MMP1 gene expression, fluorescence values of PCR products were measured by SYBR green I (Invitrogen). A reaction solution was prepared by mixing 0.2 μM primer, 50 mM KCl, 20 mM Tris / HCl pH 8.4, 0.8 mM dNTP, 0.5 U Extaq DNA polymerase, 3 mM MgCl ₂ and 1 × SYBR green in a PCR tube.

Denaturation, annealing, and polymerization were performed at 94 ° C for 30 seconds, at 58 ° C for 30 minutes, and then for 3 minutes at 94 ° C using Linegene K (BioER, China) Sec., 72 ° C for 30 sec., And the fluorescence intensity was measured after completion of each cycle.

PCR results were verified by melting curve for each result. The threshold cycle (Ct) value of each gene was normalized to the Ct value of β-actin, and the amount of change in the Ct value was compared and analyzed.

The Ct value is the number of cycles when the amount of fluorescence generated by the PCR product (the number of amplified PCR products) reaches a certain reference value above the basal value, and the amount of gene expression can be confirmed through the Ct value. The primer sequences used in the experiments are shown in Table 2 below.

Gene Forward primer Reverse primer β-actin 5-GGATTCCTATGTGGGCGACGA-3 5-CGCTCGGTGAGGATCTTCATG-3 COL1A1 5-AGGGCCAAGACGAAGACATC-3 5-AGATCACGTCATCGCACAACA-3 MMP1 5-TCTGACGTTGATCCCAGAGAGCAG-3 5-CAGGGTGACACCAGTGACTGCAC-3

The results of the measurement of the expression level according to the treatment of the extract are shown in Table 3 below. The expression level of COL1A1 mRNA was increased in a dose dependent manner, whereas the amount of MMP1 mRNA expression was decreased in a concentration dependent manner.

The higher the inhibitory effect on MMP-1 expression, the better the skin wrinkle and the anti-aging effect. The above results show that the extract promotes the production of collagen and thereby improves skin elasticity and wrinkles. .

division COL1A1 expression level MMP1 expression level Example 1 2.13 0.78 Example 2 2.25 0.73 Example 3 2.64 0.66 Example 4 2.57 0.69 Example 5 2.67 0.63 Example 6 3.10 0.47 Comparative Example 1 1.75 0.86 Comparative Example 2 1.64 0.89 Comparative Example 3 1.69 0.91 Comparative Example 4 1.82 0.83

 [Fold, β-actin normalized]

Experimental Example  3: Collagen biosynthesis induction test

The synthesis of collagen following fibroblast growth was observed.

Human dermal fibroblasts at a concentration of 1.0 × 10 ⁴ cells / mL were dispensed in a 96-well plate in an amount of 100 μL each, and cultured at 37 ° C., 5% CO ₂ and humidified conditions for 3 days. Medium 100 S (manufactured by Kurashiki Boshiki) was used a medium containing 10% by weight of LSGS (Low Serum Growth Supplement, Kurashiki Boshiki Co., Ltd.) per well.

The medium was exchanged with DMEM (Dulbecco's Modified Eagle's Medium, Sigma), and the extracts of Examples and Comparative Examples were treated at a concentration of 100 μg / mL and then cultured. The control was used in which purified water was added without treating the extract.

After the cells were cultured for 7 days, the culture solution was collected and the concentration of secreted type I collagen in the culture solution was quantified by enzyme-linked immunosorbent assay (Procollagen type I c-peptide EIA Kit, Takara Bio).

The amount of collagen in each test sample culture liquid was calculated based on the amount of type I collagen in the control (100%). The measurement results are shown in Table 4 below.

division Collagen production (%) Example 1 168.2 Example 2 171.5 Example 3 173.8 Example 4 182.2 Example 5 183.2 Example 6 191.6 Comparative Example 1 141.7 Comparative Example 2 133.4 Comparative Example 3 135.6 Comparative Example 4 136.2 Control group 101.5

Experimental Example  4 : HAS2  Gene and AQP3  Expression test

The sample powders of Examples and Comparative Examples were suspended in purified water to evaluate the skin moisturizing effect.

The degree of expression of HAS2 gene and AQP3 gene involved in skin moisturization in human keratinocytes was measured at the mRNA level.

Cultured human keratinocyte (HaCaT) under the conditions of Dulbecco's Modified Eagle's Medium (DMEM ), 10% Fatal bovine serum (FBS), 1% 37 ℃ at 100 mm / 60.1 cm culture dish along with Antibiotic-Antimycotic, 5% CO ₂ Respectively.

The keratinocytes were dispensed into 96-well plates at a concentration of 1x10 ⁶ cells / mL and cultured for 24 hours. The cells were treated with DMEM free medium and treated with the extracts of Examples and Comparative Examples diluted to a concentration of 100 μg / mL and cultured for 24 hours.

Cells were harvested, washed with cold phosphate buffered saline (PBS), and 1 ml of TRIZol reagent (Life Technologies, Inc.) was added and the total RNA extracted.

The changes in gene expression were measured by qRT-PCR (quantitative real time PCR), which binds the fluorescent substance to the DNA product amplified by polymerase chain reaction (PCR) and continuously detects the fluorescent substance.

To quantify the changes of HAS2 and AQP3 gene expression, fluorescence values of the PCR products were measured by SYBR green I (Invitrogen). A reaction solution was prepared by mixing 0.2 μM primer, 50 mM KCl, 20 mM Tris / HCl pH 8.4, 0.8 mM dNTP, 0.5 U Extaq DNA polymerase, 3 mM MgCl ₂ and 1 × SYBR green in a PCR tube.

Denaturation, annealing, and polymerization were performed at 94 ° C for 30 seconds, at 58 ° C for 30 minutes, and then for 3 minutes at 94 ° C using Linegene K (BioER, China) Sec., 72 ° C for 30 sec., And the fluorescence intensity was measured after completion of each cycle.

PCR results were verified by melting curve for each result. The threshold cycle (Ct) value of each gene was normalized to the Ct value of β-actin, and the amount of change in the Ct value was compared and analyzed.

The Ct value is the number of cycles when the amount of fluorescence generated by the PCR product (the number of amplified PCR products) reaches a certain reference value above the basal value, and the amount of gene expression can be confirmed through the Ct value. The primer sequences used in the experiments are shown in Table 5 below.

Gene Forward primer Reverse primer HAS-2 5'-TTTCTTTATGTGACTCATCTGTCTCACCGG-3 ' 5'-ATTGTTGGCTACCAGTTTATCCAAAGGG-3 ' AQP3 5'-ACCCTCATCCTGGTGATGTTTG-3 ' 5'-TCTGCTCCTTGTGCTTCACAT-3 '

The results of measurement of HAS2 and AQP3 mRNA expression levels are shown in Table 6 below, and the amount of mRNA expression was increased in a concentration-dependent manner according to the treatment of the extract. The higher the ability to promote HAS2 and AQP3 expression, the better the skin moisturizing effect.

The results suggest that the extract may enhance skin moisturization by promoting the expression of aquaporin protein and the synthesis of hyaluronic acid.

division HAS2 expression level AQP3 expression level Example 1 1.85 1.63 Example 2 1.79 1.55 Example 3 1.98 1.75 Example 4 1.94 1.68 Example 5 2.01 1.74 Example 6 2.32 2.07 Comparative Example 1 1.63 1.41 Comparative Example 2 1.56 1.34 Comparative Example 3 1.49 1.28 Comparative Example 4 1.54 1.35

 [Fold of control]

Experimental Example  5: Skin Moisture power  evaluation

The skin moisturizing power improving effect of the cosmetic compositions containing the samples of Examples and Comparative Examples was evaluated. A cream form cosmetic composition of the same composition was prepared using the samples of Examples and Comparative Examples. Each sample was formulated into cream of the same composition, except that the samples of Examples and Comparative Examples were different.

In order to measure the skin moisturizing effect of the cream form cosmetic composition, 100 subjects (20 to 40 year old female) were divided into 10 groups of 10 persons, and samples of the examples and comparative examples were included in the entire face for each group Was applied twice a day for 1 month continuously.

The skin conductivity was measured beforehand using a corneometer (CM820 courage Khazaka electronic GmbH, Germany) under constant temperature and humidity conditions (22 ± 2 ° C, humidity 50 ± 5% The skin conductivity increase rate (%) after 4 weeks was measured. The measurement results are shown in Table 7 below.

division 1 week 2 weeks 4 weeks Example 1 32 41 55 Example 2 34 45 56 Example 3 40 51 62 Example 4 34 46 58 Example 5 39 53 57 Example 6 48 57 66 Comparative Example 1 12 17 27 Comparative Example 2 11 19 26 Comparative Example 3 13 18 23 Comparative Example 4 14 20 26

Referring to Table 7, the experimental group to which the cosmetic composition of Examples 1 to 6 was applied exhibited excellent skin conductivity increase rate as compared with Comparative Examples 1 to 4. In particular, the cosmetic compositions of Examples 4 to 6 containing fermented extracts were remarkably excellent in skin conductivity increasing effect.

Experimental Example  6: Estimation of reduction effect of folliculitis

The sample powders of Examples and Comparative Examples were suspended in purified water, diluted, and applied to the skin to evaluate the effect of reducing the insect pox.

To measure sebum and to observe folliculitis, a digital light microscope (Advanced Microscopy Group Inc., USA) and slide glass, cover glass, mineral oil (Bio-Rad, USA), tweezers and the like were used.

During the course of the experiment, the sebum extracted from the nose was collected, placed on a slide glass, covered with a cover glass, photographed, and quantified by ImageJsoftware.

After 20 μL of oil was dissolved and dissolved in a slide glass containing sebum, the cover glass was covered and the presence or absence of the follicle was observed using a Digitallightmicroscope.

100 subjects were divided into 10 groups of 10 persons, 1 g of sebum was collected, and the average value of the number of folliculitis was determined. The results are shown in Table 8 below.

Since the folliculitis forms a transparent form after a short time at room temperature and is difficult to observe, the folliculitis is read in a short time to observe in the living state.

division Before use After use (1 week passed) Improvement rate (%) Example 1 12.6 9.5 24.5 Example 2 11.5 8.2 28.5 Example 3 10.7 7.7 27.9 Example 4 14.5 9.9 31.5 Example 5 12.6 8.9 29.4 Example 6 15.2 9.6 36.7 Comparative Example 1 13.5 11.7 13.6 Comparative Example 2 13.1 11.2 14.5 Comparative Example 3 14.2 11.9 16.3 Comparative Example 4 16.2 13.6 15.9

Experimental Example  7: Assessment of the inhibitory effect on survival

The sample powders of Examples and Comparative Examples were suspended in purified water and diluted, and the effect of suppressing survival of shigella feces was evaluated.

The electron microscope was attached to the CCD camera and connected to the computer. The hair follicles were separated from the hair follicles using a blade, placed on a slide glass, and treated with the extracts of Examples and Comparative Examples diluted to a concentration of 100 μg / mL.

The cover glass was covered and the movement of the body and legs of the follicle was observed under three electron microscopes.

The electron microscope was used to observe the external appearance of the folliculitis using a low magnification, and then to observe the details at increasingly high magnification and to confirm the time and measure the survival time.

In the experiment, the survival time was measured from the time when the sample was dropped to the time when the body of the fibrillar body was gradually shrunk and the movement was terminated.

division Life time Example 1 14 minutes 30 seconds Example 2 14 minutes 50 seconds Example 3 13 minutes 30 seconds Example 4 13 minutes 30 seconds Example 5 12 minutes 30 seconds Example 6 11 minutes 30 seconds Comparative Example 1 21 minutes 30 seconds Comparative Example 2 22 minutes 10 seconds Comparative Example 3 21 minutes 50 seconds Comparative Example 4 22 minutes 00 seconds

The results are shown in Table 9. Referring to Table 9, Examples 1 to 6 were superior to Comparative Examples 1 to 4 in suppressing the survival of follicular growth, and in particular, Examples 4 to 6 containing the fermentation extract were remarkably excellent in suppressing the survival of the follicular growth.

It will be understood by those skilled in the art that the foregoing description of the present invention is for illustrative purposes only and that those of ordinary skill in the art can readily understand that various changes and modifications may be made without departing from the spirit or essential characteristics of the present invention. will be. It is therefore to be understood that the above-described embodiments are illustrative in all aspects and not restrictive. For example, each component described as a single entity may be distributed and implemented, and components described as being distributed may also be implemented in a combined form.

The scope of the present invention is defined by the appended claims, and all changes or modifications derived from the meaning and scope of the claims and their equivalents should be construed as being included within the scope of the present invention.

Claims (7)

A cosmetic composition for removing folliculitis, skin moisturizing and wrinkles, comprising Cheongho extract, Ginkgo biloba extract, and Fig. The method according to claim 1,
10 to 30 parts by weight of the blueberry extract, 10 to 30 parts by weight of the ginkgo leaf extract, and 10 to 30 parts by weight of the fig extract. The method according to claim 1,
The extract is fermented extract obtained was inoculated into Lactobacillus (Lactobacillus sp.) Strains of the cosmetic composition. The method according to claim 1,
Wherein the extract is selected from the group consisting of water, an anhydrous or lower alcohol having 1 to 4 carbon atoms, acetone, petroleum ether, ethyl acetate, butyl acetate, trichloromethane, dichloromethane, chloroform, hexane and 1,3-butylene glycol Gt; cosmetic < / RTI > composition. delete delete delete