KR101908976B1 - A cosmetic composition containing lactobacillus-fermented extract for improving atopic dermatitis - Google Patents

Abstract

The present invention relates to a cosmetic composition, which comprises a cosmetic composition comprising as an active ingredient, a mixed extract consisting of a fermented extract of Lactobacillus spp. , A royal jelly extract, and a marigold extract.

Description

TECHNICAL FIELD The present invention relates to a cosmetic composition for improving atopic dermatitis containing a lactobacillus fermented extract. BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a cosmetic composition for improving atopic dermatitis,

The present invention relates to a cosmetic composition containing a lactobacillus fermented extract.

The epidermis, which is the outermost layer of the skin, protects the human body from the outside, and the stratum corneum, which is the uppermost layer of the epidermis, is an epidermal barrier. It prevents evaporation of moisture and loss of epidermis, prevention of drying of the epidermis, and prevention of bacterial, fungal, Of the human body defense mechanism to prevent the invasion of the skin is the first. The stratum corneum has stratified structures between the keratinocytes and the fat-like bricks and the plaster between them.

Atopic dermatitis is characterized by a complex disorder of skin manifesting symptoms such as strong cannibalism, dryness, severe keratinization, dandruff, erythema and swelling, skin cracking, severe dental eczema, Syndrome, which in many cases lasts for more than 10 years and is a chronic disease that causes great discomfort for long-term life. In the past, infant and childhood began with the growth of the disease gradually improved to disappear before the age of 7 to 8, mostly.

However, since the 1970s, the rate of incidence has increased year by year due to pollution caused by rapid industrialization, environmental pollution, etc. The symptoms continue to adolescence and adulthood, and the incidence of new onset after adolescence also increases remarkably. Symptoms are associated with inflammation of the skin along with strong itching, and histamine hyperalgesia is a major cause of itching. If the skin is scratched by itching, erythema or eczematous lesions will appear. In severe cases, the skin will peel off or become fizzy (skin thickening).

Although many studies and information have been presented to date on the cause of atopic dermatitis, clear theories have yet to be established regarding the etiologic etiology and effective treatments due to the conflicting research data and complexity of the disease itself.

Due to these problems, the atopic dermatitis remedy developed so far is predominantly products for adjusting the symptoms to a proper level rather than fundamental treatment such as steroid preparations, oral antihistamines, hydroxyquinolines, tar preparations and hypoallergenic moisturizers. However, none of the drugs developed so far satisfy the needs of patients.

On the other hand, cosmetics for improving atopic dermatitis require skin sedation, inflammation inhibition, bacterial inhibition, moisturizing and skin barrier maintenance effects that suppress or alleviate the itching without causing itching and skin irritation.

However, research results have been reported that the chemical preservatives (parabens, phenoxyethanol, diazolidinyl urea) used in cosmetics for the improvement of atopic dermatitis developed so far cause damage to human DNA, A case has been reported.

Accordingly, there is a growing demand for natural materials that can minimize skin irritation and side effects by minimizing the use of chemical substances, and various studies on the development of cosmetics for improving atopic dermatitis, which have excellent biostability and skin soothing effect, .

Accordingly, the present inventors have developed an eco-friendly skin external preparation product that can contain only natural ingredients corresponding to EWG 1 grade and can safely use sensitive skin.

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made keeping in mind the above problems occurring in the prior art, and an object of the present invention is to provide a cosmetic composition containing a natural raw material as an active ingredient.

According to one aspect of the present invention, there is provided a cosmetic composition comprising, as an active ingredient, a mixed extract consisting of a fermented extract of Lactobacillus spp. , An extract of Angelica gigas Nakai, and a marigold extract.

In one embodiment, the Lactobacillus fermentation extract may be obtained by fermenting pomegranate extract.

In one embodiment, the Lactobacillus fermentation extract may be obtained by inoculating Lactobacillus plantarum .

In one embodiment, the mixed extract may comprise 20 to 60 parts by weight of the lactobacillus fermentation extract, 10 to 20 parts by weight of the Young's Silk Extract, and 10 to 20 parts by weight of the march extract.

In one embodiment, the cosmetic composition may comprise from 0.5 to 10.0% by weight of the mixed extract.

In one embodiment, the mixed extract may be extracted with one or more solvents selected from the group consisting of purified water, lower alcohols having 1 to 4 carbon atoms, acetone, ethyl acetate, butyl acetate, and 1,3-butyleneglycol .

In one embodiment, the cosmetic composition may be for improving atopic dermatitis having antioxidant, anti-inflammatory, and antibacterial activity.

In one embodiment, the cosmetic composition is a composition comprising a softening agent, a convergent lotion, a nutritional lotion, a lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray, ≪ / RTI >

The cosmetic composition according to one aspect of the present invention is effective for the skin health of the user by interacting with the active ingredients contained in the mixed extract, and can maximize the skin soothing effect and the atopic dermatitis improvement effect.

In particular, the natural raw material applied to the cosmetic composition of the present invention is a raw material corresponding to the EWG 1 grade and has no harmful effect on the human body, and sensitive skin can be applied safely.

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 provides a cosmetic composition comprising, as an active ingredient, a mixed extract consisting of a fermented extract of Lactobacillus spp. , A sweet potato extract, and a marigold extract.

The cosmetic composition of the present invention contains an extract of natural origin as an active ingredient and has excellent antioxidative, anti-inflammatory and antibacterial activities, thereby alleviating skin irritation and effectively improving atopic dermatitis. In particular, since the mixed extract acts on the skin at the same time, the effect of improving sedative and atopic dermatitis can be remarkably increased due to the synergistic effect.

In particular, since the cosmetic composition contains only EWG level 1 components, it satisfies both functionality and safety, and is easy to apply to sensitive skin because it is not harmful.

EWG is a cosmetic ingredient harmful grade announced by EWG (Environmental Working Group), a nonprofit environmental NGO in the United States. EWG carefully examines the harmfulness of cosmetic ingredients and classifies the hazard level from 1 to 10, and 1 Class 2 and Class 2 are classified as safety class.

The Lactobacillus fermentation extract means a mixture of fermentations obtained by fermenting a strain of Lactobacillus spp. On a medium containing an energy source such as skim milk powder. The fermented product can be produced by a conventional lactobacillus fermentation method. The lactic acid bacteria fermentation method is apparent to those skilled in the art to which the present invention belongs.

The energy source may be cow milk, goat milk or horse milk and whole milk, skim milk or powder thereof, and can be used in powder form.

The strain of the genus Lactobacillus may be selected from the group consisting of Lactobacillus pentosus , Lactobacillus brevis , Lactobacillus plantarum , Lactobacillus casei or Lactobacillus acidophilus ( Lactobacillus acidophilus ), but is not limited thereto.

On the other hand, the lactobacillus fermentation extract can be obtained by fermenting pomegranate extract and inoculating Lactobacillus plantarum .

The term " fermentation " means a process of decomposing an organic substance using an enzyme contained in the microorganism. It has been reported that a raw material that has undergone fermentation by the above-mentioned 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, as the fermentation process proceeds, the particles become smaller and the toxicity such as heavy metals is decomposed, so that the absorption rate is good, and skin troubles and allergic side effects can be alleviated.

The above-mentioned " 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 pomegranate extract to a certain concentration in a solvent, preferably purified water, , That is, by inoculating and fermenting a strain of Lactobacillus sp .

In particular, the antioxidative and skin elasticity-improving effects of the pomegranate extract can be significantly enhanced by the fermentation of the lactobacillus strain.

The "pomegranate ( Punicagranatum L. )" is a fruit of the pomegranate family ( Punicaceae ), also called aphid pomegranate, anthrax or sweetpot. The major components of the pomegranate are saccharides (glucose fructose), about 40%, and the organic acid of pomegranate contains about 1.5% of citric acid, which gives sour taste, to purify and circulate blood. The pomegranate is rich in minerals such as calcium, potassium, magnesium, zinc, and iron that perform important functions in our body. In addition, pomegranate contains vitamins such as vitamin B1, vitamin B2 and vitamin C, which provide various nutrients to control various functions and prevent disease in the body.

Is a fruit of Rosa multiflora var. Platyphylla , which is commonly used in the treatment of arthritis and toothache. Petal is used as a lotion, and its fruit, Youngsil, is known to be a medicinal herb that has excellent therapeutic effects on women's menstrual cramps, menstrual irregularities, constipation, nephritis, cystitis, In addition, it contains various active ingredients and exhibits various skin improving effects including atopic improvement.

The above-mentioned " Portulaca oleracea " is an annual plant belonging to the leguminous plants, leguminous plants and leggings of the leguminous plants of the leggings ( Portulacaceae and Portulacaceae ). It is also called leggings, five legs, long legs, The mashin is known to have an effect on detoxification, diuretic and dysentery in pharmacological activity, and it is known to be effective on detoxification, diuretic and dysentery, It is also called horsemeat.

The mixed extract may exhibit excellent skin sedation and atopic dermatitis improving effect due to interactions between active ingredients derived from each raw material. The combined extracts can be applied to the skin together with the various components contained in the respective extracts, thereby increasing the absorption rate and maximizing the effect of improving skin sedation and atopic dermatitis.

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 mixed extract may be extracted with at least one solvent selected from the group consisting of purified water, a lower alcohol having 1 to 4 carbon atoms, acetone, ethyl acetate, butyl acetate, and 1,3-butylene glycol, It may be ethanol at 90% concentration (v / v).

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 .

Particularly, the polarity of water and ethanol is different, so that the effective ingredient to be extracted according to each polarity can be changed, and 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 the ethanol exceeds 90%, an adequate yield may not be realized. If the concentration is less than 60%, the effective ingredient showing skin improving effect may not be extracted sufficiently.

The mixed extract is prepared by a conventional method such as reflux circulation extraction, pressure extraction, and ultrasonic extraction for about 1 to 24 hours with a solvent having a volume of 8 to 12 times the weight of the raw material after washing each material with water, drying and pulverizing Extraction and filtration. 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 comprise from 0.5 to 10.0% by weight of the mixed extract. If the amount of the mixed extract is less than 0.5% by weight, the effect of improving the atopic dermatitis by the mixed extract may not be realized properly. If it exceeds 10.0% by weight, the specific gravity of the other additives required for commercialization is decreased, .

The mixed extract may be composed of 20 to 60 parts by weight of the lactobacillus fermentation extract, 10 to 20 parts by weight of the spiritual extract, and 10 to 20 parts by weight of the march extract.

In the case where the content difference between the active ingredients constituting the mixed extract is out of the above range, the synergistic effect due to the interaction between the active ingredients may not be properly realized. Therefore, The ratio can be appropriately controlled.

The cosmetic composition may be used for skin soothing or atopic dermatitis improvement and may be used for softening the skin such as a softening agent, a convergent lotion, a nutritional lotion, a lotion, a nutrition cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, And a powder.

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.

Production Example 1: Preparation of fermented extract using Lactobacillus strain

The Lactobacillus plantarum strain was inoculated into a defatted milk powder medium and incubated at 37 DEG C for 3 days, followed by centrifugation to obtain a supernatant. The pH of the recovered supernatant was adjusted to 6.5 and then lyophilized to obtain a fermented product. The composition of the skim milk powder medium is shown in Table 1 below.

ingredient Concentration (g / L) Skim milk powder 100 glucose 20 Yeast extract 3 peptone 3 Distilled water One

Production Example 2: Preparation of pomegranate lactobacillus fermentation extract

The fermentation medium (glucose 1.5% (v / v), sugar 1.5% (v / v), starch syrup 1.5% (v / v), yeast extract 0.3%) containing glucose, sugar, syrup and yeast extract v / v)) was used to prepare a fermented extract of Lactobacillus plantarum .

To the medium (10 mL) was added 10 mg of the dried product of Lactobacillus plantarum strain and pre-cultivation was performed. The pomegranate extract was added to a medium having the same composition as the fermentation medium, and the pre-culture solution was inoculated and fermented.

Lactobacillus plantarum strain was cultured at 36.5 ℃ for 36 hours and glucose and sucrose contents were controlled to increase the growth rate of the strain. After the fermentation was completed, the mixture was centrifuged and filtered to obtain a pomegranate lactobacillus fermentation extract.

Production Example 3: Preparation of natural extract (hot water extraction)

Youngsil, and Mahi-Hyun were washed with water and then dried and pulverized at room temperature to obtain 100 g of crude pulverized products.

100 g of each of the pulverized products was immersed in a 10-fold volume of purified water and reflux-extracted at 80 DEG C for 12 hours.

The extract was filtered through a fine filter and repeatedly extracted three times in the same manner for the remaining raw materials and then cooled at room temperature. Each of the extracts was concentrated under reduced pressure at 30 ° C and lyophilized to obtain a solid.

Production Example 4: Preparation of natural extract (ethanol extract)

Each extract was obtained in the same manner as in Production Example 3, except that an ethanol aqueous solution (70% mass concentration) was used as an extraction solvent. Each extract was concentrated under reduced pressure and then lyophilized to obtain a solid.

Example 1: Mixed extract (hot water extraction) Samples

The extracts of Preparation Example 1 and Preparation Example 3 obtained by lyophilization were mixed in the same ratio. 10 g of the mixed complex extract was used as a sample of Example 1.

Example 2: Mixed extract (hot water extraction) Samples

The extracts of Preparation Example 2 and Preparation Example 3 obtained by lyophilization were mixed in the same ratio. 10 g of the above-mentioned mixed complex extract was used as a sample of Example 2.

Example 3: Mixed Extract (Ethanol Extraction) Samples

The extracts of Preparation Example 1 and Preparation Example 4 obtained by lyophilization were mixed in the same ratio. 10 g of the above-mentioned mixed complex extract was used as a sample of Example 3.

Example 4: Mixed Extract (Ethanol Extraction) Samples

The extracts of Preparation Example 2 and Preparation Example 4 obtained by lyophilization were mixed in the same ratio. 10 g of the mixed complex extract was used as a sample of Example 4.

Comparative Example 1: Lactobacillus fermentation extract sample

10 g of the lactobacillus fermentation extract of Preparation Example 1 obtained by lyophilization was used as a sample of Comparative Example 1.

Comparative Example 2: Pomegranate Lactobacillus extract sample

10 g of the pomegranate Lactobacillus extract of Preparation Example 2 obtained by lyophilization was used as a sample of Comparative Example 2.

Comparative Example 3: Extract of Young-Sil (hot-water extract) Samples

10 g of the Young's Silk Extract of Preparation Example 3 obtained by lyophilization was used as a sample of Comparative Example 3.

COMPARATIVE EXAMPLE 4: Sucrose extract (hot water extraction) Samples

10 g of the marigold extract of Preparation Example 3 obtained by lyophilization was used as a sample of Comparative Example 4.

COMPARATIVE EXAMPLE 5: Extracts of Young Sil and Mashin (ethanol extract) Samples

5 g of the Young's Silk Extract of Preparation Example 3 obtained by freeze-drying and 5 g of the mash extract were mixed to prepare a sample of Comparative Example 5.

Comparative Example 6: Young silk and Horseradish extract (ethanol extract) Samples

5 g of the crude extract of Preparation Example 4 obtained by freeze-drying and 5 g of the crude extract were used as a sample of Comparative Example 5.

Experimental Example 1: Evaluation of free radical scavenging activity

The extracts of Examples and Comparative Examples were suspended in purified water (concentration 10%) to evaluate free radical scavenging activity.

The DPPH assay measures absorbance at 540 nm to the extent that the inhibitor decolorizes the stable radical DPPH (2,2-Diphenyl-1-picrylhydrazyl radical).

As a control, free radical scavenging activity was clearly compared using dibutylhydroxy toluene (BHT). Dibutylhydroxytoluene is a colorless crystal or a white crystalline powder, which is excellent in antioxidative effect and is widely used as an antioxidant for edible and maintenance foods.

The same experiment was repeated 3 times for the accuracy of the experiment, and the results are shown in Table 2 below.

[% of control] division DPPH radical scavenging activity (%) Example 1 131.7 Example 2 135.6 Example 3 134.6 Example 4 141.7 Comparative Example 1 104.2 Comparative Example 2 108.4 Comparative Example 3 115.1 Comparative Example 4 112.8 Comparative Example 5 119.4 Comparative Example 6 122.6 Control group 147.9

Referring to Table 2, the free radical scavenging activity of the samples of Examples 1 to 4 was increased as the concentration was increased, and the free radical scavenging activity was significantly better than that of Comparative Examples 1 to 6. [

The higher the free radical scavenging activity, the better the antioxidative effect. In particular, the sample of Example 4, in which the pomegranate lactobacillus fermentation extract and the ethanol extract were simultaneously applied, was more excellent in antioxidative effect than those of Examples 1 to 3.

Experimental Example 2: Evaluation of anti-inflammatory activity

The anti - inflammatory activity of EAME was evaluated by analyzing the inhibitory effect of NO production on EAME in mouse macrophage RAW 264.7 cells stimulated with Lipopolysaccharide (LPS).

Raw 264.7 cells, a Murine macrophage cell line, were purchased from Korean Cell Line Bank and cultured in DMEM medium containing 100 units / mL penicillin-streptomycin and 10% fetal bovine serum (FBS) at 37 ° C in a 5% CO ₂ incubator And subculture was performed once every 2 to 3 days.

Raw 264.7 cells (3 × 10 ⁵ cells / mL) were cultured for 18 hours, and the extract samples (concentration 10%) and LPS (1 μg / mL) of the examples and comparative examples were simultaneously treated for 24 hours, NO was measured in the form of NO ₂ ^- present in cell culture medium using Griess reagent. The negative control group was not treated with LPS, and the positive control group was treated with LPS alone to measure the amount of NO produced.

Cell culture supernatants (100 μL) and Griess reagent (1% w / v sulfanilamide, 0.1% w / v naphtylehtylenediamine in 2.5% (v / v) phosphoric acid) were mixed and equilibrated in a dark room for 10 min. absorbance (540 nm) was measured using a reader. Standard concentration curves were obtained by serial dilution of sodium nitrite (NaNO ₂ ) (10-100 μM). The measurement results are shown in Table 3 below.

division NO production (μM) Example 1 15.47 Example 2 13.72 Example 3 13.52 Example 4 8.38 Comparative Example 1 29.53 Comparative Example 2 24.94 Comparative Example 3 26.61 Comparative Example 4 28.23 Comparative Example 5 22.52 Comparative Example 6 20.12 LPS (-) 6.64 LPS (+) 37.21

Referring to Table 3, the samples of Examples 1 to 4 were superior to the samples of Comparative Examples 1 to 6 in inhibiting nitric oxide (NO) formation, and the antioxidative and anti-inflammatory activities were remarkably improved. In addition, the sample of Example 4, to which the pomegranate lactobacillus fermentation extract and the ethanol extract were applied at the same time, was further enhanced in anti-inflammatory activity as compared with Examples 1 to 3.

Experimental Example 3: Antimicrobial activity evaluation

The antibacterial activity of the samples obtained in Examples and Comparative Examples was evaluated. Propionibacterium acnes KCTC 3314 was used as a test strain and triclosan, a chemical synthetic antimicrobial, was used as a positive control.

Propionibacterium acnes were streaked on tryptic soy agar medium containing 5% sheep blood and cultured for 2 to 3 days to prepare McFaland nephleometer standards 0.5 (1% Barum chloride (0.05 ml) + 1% Sulfuric acid (9.95 ml), the bacterial concentration was 1 × ^{10 8} cells / mL.

Each sample was diluted in 2 ml of liquid BHI medium at intervals of 100 ppm from 0 to 2000 ppm, and 20 μl of 1x10 ⁸ cells / ml of the bacterial solution was inoculated and cultured for 48 hours. The minimum growth inhibition concentration was defined as the lowest growth inhibitory concentration (MIC) based on turbidity, and the results are shown in Table 4 below.

division Minimum growth inhibitory concentration MIC (PPM) Example 1 730 Example 2 620 Example 3 650 Example 4 410 Comparative Example 1 1750 Comparative Example 2 1550 Comparative Example 3 1450 Comparative Example 4 1700 Comparative Example 5 1300 Comparative Example 6 1150 Triclosan 400

Referring to Table 4, the antibacterial effect of the samples of Examples 1 to 4 was somewhat inferior to that of the positive control triclosan, but the antimicrobial effect was remarkably superior to those of Comparative Examples 1 to 6.

According to the evaluation results, when the pomegranate lactobacillus extract and the ethanol extract were applied, the antimicrobial effect against the dicycles was further improved.

Experimental Example 4: Evaluation of improving effect of atopic dermatitis

The effect of improving the atopy of the cosmetic composition containing the samples obtained in the Examples and Comparative Examples was evaluated. Each of the above samples was formulated into cream of the same composition, and samples of Examples 1 to 4 and Comparative Examples 1 to 5 were different.

100 selected women aged 20 to 60 years old were evaluated for the degree of improvement of atopic dermatitis by applying formulations of Examples and Comparative Examples.

100 women who had similar dermatitis symptoms were divided into 10 groups each consisting of 10 individuals. The formulated cream was applied to the whole face for 12 weeks twice a day at a temperature of 24 to 26 DEG C and a humidity of 75%.

At the end of the test, subjects were asked to fill out a questionnaire and conducted a subjective efficacy evaluation. In the questionnaire, the symptoms of atopic dermatitis were classified into three categories: "leeches", "dryness", "keratinization", "dandruff", "erythema", "swelling", "skin crack" . When the symptom was present, the degree of improvement was evaluated for each symptom, and the improvement was evaluated by individuals to evaluate the improvement effect of atopic dermatitis symptoms. The evaluation results are shown in Table 5 below.

sample Significant improvement (persons) Some improvement (persons) No improvement (persons) Deterioration (persons) Example 1 6 3 One - Example 2 7 2 One - Example 3 8 One One - Example 4 9 One - - Comparative Example 1 2 5 2 One Comparative Example 2 3 4 2 One Comparative Example 3 3 3 3 One Comparative Example 4 3 2 3 2 Comparative Example 5 4 3 2 One Comparative Example 6 5 3 One One

As shown in Table 5, the samples 1 to 4 were evaluated as having excellent atopic dermatitis improving effect as compared with the samples of Comparative Examples 1 to 6.

In addition, the sample of Example 2 to which the pomegranate lactobacillus fermentation extract was applied had a better effect of improving atopic dermatitis compared to Example 1. The sample of Example 4 to which ethanol extract was applied showed better skin improvement effect than that of Example 2 to which hot water extract was applied Was significantly improved.

That is, the samples of Examples 3 and 4 by ethanol extraction significantly improved the atopic dermatitis and the skin soothing effect as compared with Examples 1 and 2, respectively.

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 (8)

A mixed extract consisting of a fermented extract of Lactobacillus spp. , An Aspergillus oryzae extract, and a marjoram extract as an active ingredient,
Wherein the lactobacillus fermentation extract is obtained by fermenting pomegranate extract. delete The method according to claim 1,
Wherein the lactobacillus fermentation extract is obtained by inoculating Lactobacillus plantarum . The method according to claim 1,
Wherein the mixed extract comprises 20 to 60 parts by weight of the lactobacillus fermentation extract, 10 to 20 parts by weight of the farinaceutic extract, and 10 to 20 parts by weight of the aspartame extract. The method according to claim 1,
And 0.5 to 10.0% by weight of the mixed extract. The method according to claim 1,
Wherein the mixed extract is extracted with at least one solvent selected from the group consisting of purified water, a lower alcohol having 1 to 4 carbon atoms, acetone, ethyl acetate, butyl acetate, and 1,3-butylene glycol. 7. The method according to any one of claims 1 to 6,
Which is effective for improving atopic dermatitis having antioxidant, anti-inflammatory, and antibacterial activity. 7. The method according to any one of claims 1 to 6,
A cosmetic composition formulated with at least one selected from the group consisting of softening lotion, astringent lotion, nutritional lotion, lotion, nutritional cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray, .