KR101940658B1 - A cosmetic composition for slimming comprising ilex paraguariensis leaf extract - Google Patents

Abstract

The present invention provides a cosmetic composition comprising a matte extract as an active ingredient.

Description

TECHNICAL FIELD The present invention relates to a cosmetic composition for slimming, which contains an extract of Mattera as an active ingredient,

The present invention relates to a functional cosmetic composition containing a natural extract as an active ingredient.

Obesity generally refers to abnormal hypertrophy of fat adipose tissue below the skin layer. This phenomenon is generally caused by the fact that the calories of the foods to be consumed are more than necessary but that when the calories to be energized and digested are small, the excess calories are accumulated in the form of fat in the body. As a result, the obesity phenomenon causes the body to become fat and lose its physical beauty due to the fat thickness accumulated below the skin layer, and causes various adult diseases such as hypertension and diabetes. Therefore, .

Particularly, cellulite is a fat component that accumulates excessively in a specific region, and it causes a phenomenon such as sagging, elastic deformation and obesity in a specific region such as a face, a waist, a thigh or a chest. In the human body, fat is accumulated in the adipocyte, an adipocyte, in the form of triacylglycerol, so that triacylglycerol must be removed in order to decompose the fat.

In general, about 20 billion fat cells are present in the human body and play a role in accumulating or releasing energy in mammals' in vivo. In this cell, there is a complicated control principle for the accumulation and release of energy. When the supply is much higher than the demand of energy, it is stored as neutral fat in the adipocyte and decomposed into free fatty acid and glucose when energy is depleted . Obesity is caused by the accumulation of excessive energy due to the imbalance of the process, which is attributed to the increase in the size of fat cells or the increase in the number of fat cells.

The differentiation process of adipocytes is constantly converted from adipose precursor cells to adipocytes throughout life, which occurs through many external stimuli such as hormonal induction and complex gene regulation processes. Among the external signals that induce the differentiation of adipocytes, insulin is the most typical hormone and plays a pivotal role in the regulation of adipocyte metabolism. Most of the lipolysis-promoting substances conventionally known are caffeine, theophylline, Are the same methylxanthine substances. These substances exhibit lipolytic activity through promotion of adenylatecyclase, which is an enzyme closely related to lipolysis in adipocytes, or inhibition of phosphodiesterase.

However, they are known to be addictive substances, and it is known that there are side effects such as inducing bronchodilation or exacerbating the premenstrual syndrome phenomenon, and there is no clear verification of its safety. In addition, skin elasticity and hormone changes may cause skin aging and troubles.

Meanwhile, 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.

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 to solve the above problems occurring in the prior art, and it is an object of the present invention to provide a cosmetic preparation composition which promotes the decomposition process of neutral fat accumulated in fat cells, .

According to one aspect of the present invention, there is provided a cosmetic composition comprising a matte extract as an active ingredient.

In one embodiment, it may further comprise ginseng extract, turmeric extract, pepper seed extract.

In one embodiment, the extract is selected from the group consisting of Bifidobacterium spp . ) Strain of the present invention.

In one embodiment, the Bifidobacterium spp . ) Strain Bifidobacterium Bifidobacterium bonus (Bifidobacterium bifidum ).

In one embodiment, 20 to 70 parts by weight of the matte extract, 10 to 20 parts by weight of the ganoderma extract, 10 to 20 parts by weight of the turmeric extract, and 10 to 20 parts by weight of the pepper seed extract may be included.

In one embodiment, the 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 slimming, skin moisturizing or skin cleansing.

In one embodiment, the cosmetic composition is selected from the group consisting of softening longevity, convergent lotion, nutritional lotion, lotion, nutritional cream, massage cream, essence, cleansing cream, cleansing foam, cleansing water, pack, spray, Or more.

The cosmetic composition according to one aspect of the present invention is excellent in the decomposition effect of triglycerides present in adipocytes by containing a matte extract as an active ingredient and thus can be effectively used as a cosmetic for slimming which reduces the subcutaneous fat layer But can be used as skin moisturizing or soothing cosmetics. In addition, when one or more components are further combined with the extract of the matte, the slimming effect can be remarkably improved due to the synergy effect of each component.

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 can be applied safely to sensitive skin without harmfulness to human body.

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.

According to one aspect of the present invention, there is provided a cosmetic composition comprising a matte extract as an active ingredient.

The cosmetic composition of the present invention not only can reduce the subcutaneous fat layer based on the lipolytic effect but also can achieve skin moisturizing or skin soothing effect by including an extract derived from natural origin as an effective ingredient. Particularly, since the mixed extract acts on the skin simultaneously, slimming, skin moisturizing or soothing effects can be remarkably increased due to the synergistic effect.

The " comprising as an active ingredient " means an effective amount of an effective amount which can exhibit a skin improving effect, for example, a degree of fat dissolving effect associated with weight loss efficacy, skin moisturizing, skin irritation alleviation, It can mean.

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 "Do tape (Ilex paraguariensis leaf "is a plant of about 6 meters in height belonging to a gambling tree family originating from the subtropical highlands of Brazil, Paraguay, Uruguay and Argentina. The matte contains caffeine and smells good, especially called 'car-eating salad' and is widely consumed in South America. Mate is rich in minerals such as iron, calcium, and magnesium because it grows in rich minerals such as calcium and iron. Especially, the iron has high iron content and has iron content which is five times that of green tea leaves.

In addition, the mate has a function of lowering the edible state by allowing a feeling of fullness to be felt within a short period of time when consumed, so that chlorate acid contained in the matte itself can not only prevent the absorption of fat but also decompose fat An excellent slimming effect can be realized.

Meanwhile, the cosmetic composition may further comprise ginseng extract, turmeric extract, and pepper seed extract.

The " Ganoderma lucidum ) "is a kind of mushroom native to the dead wood and stump of broad-leaved trees and is distributed north of the temperate zone of the Northern Hemisphere. Gnocchi mushroom is a one-year-old mushroom with polished surface on both sides and gut. Some are round with yunmun and occasionally elliptical. The front face is yellowish white at first, but it grows from reddish brown to dark brown from the first grown part. The back side is yellowish white and there are a lot of bays. The stand is slightly curved with the same color as the surface of the umbrella, and some large ones have wrinkles of 30 cm and lengths of over 20 ㎝. It collects in autumn, unlike other edible mushrooms, it does not rot and remains polished even after it is dead.

The Ganoderma lucidum exhibits organs such as the heart, lungs, and liver, strengthens the muscles and bones, and strengthens the energy. The various active ingredients of Ganoderma lucidum provide skin moisturization and skin irritation mitigation by ultraviolet rays. .

The "turmeric (Curcuma longa ) "is divided into Ulguk and turmeric depending on the area where medicinal herbs are used as originals. The vegetation type, weakness, etc. described in the old consent document use the roots of the plant roots, while the roots of the roots utilize stem roots of the plant, and the taste is much higher than that of the roots, and the color is darker than the roots. In addition, small spiny fluffy flowers on the back side of the leaves grow in pink in spring, and tropical Asia is native to the country, and turmeric plants are not native to the country. They are perennial herbaceous plants grown in tropical regions and southern China. to be.

Curcumin, which is a major physiologically active ingredient contained in turmeric, is used as an antitumor agent, antioxidant, anti-amyloid, and the like. And it has an effect of inhibiting the inflammation reaction, so that the skin soothing effect can be realized.

The above-mentioned " Piper nigrum seed " is a seed of pepper. The pepper belongs to the evergreen plant of the dicotyledonous plant, the pepper tree, and the southern part of India, and is distributed in Korea, Indonesia, and West Indies Islands. The fruit of the pepper is green when it is less ripe and turns red when it is ripe. The pepper seeds have a warm property, have an anticancer effect, an antioxidant effect, and an effect to smooth blood circulation, thereby cleansing the skin. When applied to a cosmetic composition, the skin can be moisturized and the skin soothing effect can be realized.

The ginseng extract, turmeric extract, and pepper seed extract can exhibit excellent slimming effect and skin improving effect due to interaction between active ingredients derived from each raw material. The extract, turmeric extract, and pepper seed extract can be increased in absorption rate by applying various components contained in each extract together to the skin. In particular, since the extract has skin moisturizing and skin soothing effects, it is possible to diversify the functionality of the cosmetic composition and to maximize the effects of lipid decomposition and body fat reduction by the extract of the maturation.

The cosmetic composition may comprise 20 to 70 parts by weight of the matte extract, 10 to 20 parts by weight of the ganoderma extract, 10 to 20 parts by weight of the turmeric extract, and 10 to 20 parts by weight of the pepper seed extract.

In the case where the content deviation between the respective active ingredients constituting the extract is out of the above range, a synergistic effect due to interaction between the active ingredients may not be properly realized. Therefore, considering the working environment and the quality of the final product, Can be appropriately controlled.

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 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, % Concentration (v / v) of ethanol.

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 Can

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 extract is washed with water, dried and pulverized, and extracted and 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, Followed by 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 >

On the other hand, the above extract is classified into Bifidobacterium spp . ) Strain of the present invention.

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 Bifidobacterium spp . ) Is a microorganism which mainly utilizes oligosaccharides containing aminosugars and promotes the digestion and absorption of proteins by inhibiting the development of harmful bacteria in the intestines and provides a vitamin group to the body. The Bifidus fermented extract has a natural moisturizing factor NMF, natural moisturizing factor), which is a precursor of profilaggrin, is converted into a natural moisturizing factor, thereby improving skin moisturization, imparting moisture and gloss to the skin, and achieving various skin improving effects. In addition, the polysaccharide component rich in the fermented extract can inhibit water evaporation in the skin surface.

The bifidus strain may be Bifidobacterium breve), Bifidobacterium Bifidobacterium bonus (Bifidobacterium may be bifidum), Bifidobacterium ronggeom (Bifidobacterium longum), or Bifidobacterium inpeon tooth (Bifidobacterium infantis) may be a, preferably Bifidobacterium bipyridinium bushes (Bifidobacterium bifidum).

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, preferably in purified water, and then adding a fermentation microorganism, And then fermented.

The cosmetic composition may be used for slimming, skin moisturizing or skin cleansing.

The term " slimming " means stimulating or decomposing adipose cells to release and simultaneously reduce the fat mass that is gathered in the body. In other words, not only for the purpose of reduction of fatigue, but also to loosen the mass of fat clusters to trim the body line, has the effect of increasing skin elasticity.

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 above-mentioned " skin soothing " means all actions that inhibit the inflammation activity that causes irritation to the skin, thereby suppressing troubles and the like. The skin soothing effect may be accompanied by an additional skin improving effect in various aspects such as a skin redness and erythema improvement effect, a skin irritation reduction effect, and a trouble improving effect such as acne.

The cosmetic composition may be at least one selected from the group consisting of softening longevity, nutritional lotion, moisture cream, nutritional cream, massage cream, essence, ampoule, gel, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray and powder Can be formulated.

The cosmetic composition may be formulated by a conventional method in the art. 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 extract

Matsutake, manji, turmeric, and black pepper were washed with water, completely dried at room temperature, and pulverized to obtain 100 g of each pulverized product.

100 g of each pulverized product was immersed and extracted with a 10-fold volume of an aqueous ethanol solution (70% mass concentration) as a solvent at a temperature of 70 ° C for 12 hours.

The extract was filtered with 250 mesh and 0.5 mu m filters and then extracted three times in the same manner for the remaining raw materials. The extract was concentrated under reduced pressure at 30 DEG C and lyophilized to obtain a solid fraction.

Manufacturing example  2: Preparation of fermented extract

Each extract obtained in Preparation Example 1 was inoculated into a fermentation medium (glucose 1.5% (v / v), sugar 1.5% (v / v), starch syrup 1.5% (v / v), starch syrup, yeast extract, / v), and yeast extract (0.3% (v / v)). Bifidobacterium longum ) was prepared.

10 mg of Bifidobacterium bifidobacterium strain was added to the medium (10 mL) and pre-cultivated. Artichoke leaf extract was added to a medium having the same composition as the fermentation medium, and the preculture was inoculated and fermented.

Bifidobacterium bifidum strain was cultured at 36.5 ℃ for 36 hours and glucose and sugar content were controlled to increase the growth rate of the strain during culture. After the fermentation, the fermented extracts of the respective extracts were obtained by centrifugation and filtering.

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 raw materials were mixed at predetermined ratios as shown in Table 1 below to verify skin improvement effect.

[Content (parts by weight)] division Example
One Example
2 Example
3 Example
4 Comparative Example
One Comparative Example
2 Comparative Example
3 Comparative Example
4 Comparative Example
5 Comparative Example
6 Matte extract 70 40 - - - - - - - - Ganoderma extract - 10 - - 70 - - - - - Turmeric extract - 10 - - - 70 - - - - Pepper seed extract - 10 - - - - 70 - - - Mate
Fermented extract - - 70 40 - - - - - - wisdom
Fermented extract - - - 10 - - - 70 - - curcuma
Fermented extract - - - 10 - - - - 70 - Mr. Fuchu
Fermented extract - - - 10 - - - - - 70

Experimental Example  1: Skin safety test

MTT assay experiments were performed on fibroblasts (HDF), melanocytes (B16F10), and keratinocytes (HaCaT) to verify skin safety for the samples of the Examples and Comparative Examples.

The samples of Examples and Comparative Examples were each suspended in purified water at a concentration (50 mg / L), 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 was added. The cells were cultured at 37 ° C and 5% CO ₂ for 24 hours. 100 L of DMSO (dimethyl sulfoxide) was added to each well, stirred for 10 minutes, and absorbance was measured at 540 nm.

As a result of measurement, cytotoxicity was not confirmed at all concentrations of each sample. The results suggest that the extract is not only harmless to human body but also has excellent human safety.

Experimental Example  2: Collagen biosynthesis induction test

It was confirmed that collagen synthesis was promoted by the growth of fibroblasts.

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 samples of the above-mentioned Examples and Comparative Examples were treated at a concentration of 50 mg / L and then cultured. As a control, acetic acid (50 μg / mL) was used as a positive control.

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 the culture medium of each test sample was calculated based on the amount of type I collagen in the control (100%). The measurement results are shown in Table 2 below.

[% of control] division Collagen production (%) Example 1 142.8 Example 2 145.6 Example 3 144.8 Example 4 152.4 Comparative Example 1 112.6 Comparative Example 2 110.6 Comparative Example 3 107.4 Comparative Example 4 110.3 Comparative Example 5 112.8 Comparative Example 6 114.6 Positive control group 132.7

Experimental Example  3: skin moisturizing effect test

Samples of the Examples and Comparative Examples were suspended in purified water and diluted to a concentration of 50 mg / L 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 plated at a density of 1.0 x ¹⁰ < ⁶ > cells / mL into 96-well plates and cultured for 24 hours. After replacing with DMEM free medium, the samples of Examples and Comparative Examples diluted to a predetermined concentration were treated 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 3 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 amounts are shown in Table 4 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 above results can effectively promote the expression of the aquaporin protein and the synthesis of hyaluronic acid by interacting with the extracts of the matte, ganoderma, cadmium, and pepper seed extracts and their fermentation extracts, Suggesting that the effect can be improved.

[% of control] division HAS2 expression level AQP3 expression level Example 1 124.1 127.6 Example 2 127.2 129.7 Example 3 126.7 125.7 Example 4 137.5 138.6 Comparative Example 1 110.7 112.4 Comparative Example 2 112.6 103.7 Comparative Example 3 102.9 107.2 Comparative Example 4 113.8 109.5 Comparative Example 5 103.6 113.7 Comparative Example 6 110.5 108.5

Experimental Example  4: 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 5 below.

division NO production (μM) Example 1 14.71 Example 2 10.67 Example 3 8.46 Example 4 7.21 Comparative Example 1 28.15 Comparative Example 2 27.67 Comparative Example 3 26.95 Comparative Example 4 27.33 Comparative Example 5 25.67 Comparative Example 6 27.11 LPS (-) 6.17 LPS (+) 37.24

Referring to Table 5, 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 anti-inflammatory activity was remarkably improved. In addition, the sample of Example 2 to which the mixed extract further containing the ganoderma extract, the turmeric extract, and the pepper seed extract was applied, and the sample of Example 4 to which the mixed fermentation extract was applied showed a higher anti-inflammatory activity than the single extracts of Examples 1 and 3 .

Experimental Example  5: Assessment of Triggering Effects of Neutral Fat Decomposition in Adipocytes

3T3-L1 cells, a fibroblast cell line of mice, were cultured in DMEM containing 10% fetal bovine serum (FBS) (Dulbecos modified eagles medium, GIBCO BRL, Life Technologies) And then attached to a well culture plate at 1 × 10 ⁵ cells / well. After 2 days, the cells were replaced with fresh DMEM (containing 10% FBS) medium and cultured for 2 days. The cultured cells were again induced to differentiate into DMEM (containing 10% FBS) containing 1 μg / ml insulin, 0.5 mM IBMX and 0.25 μM dexamethasone, and after 2 days, insulin was again contained Gt; EMEM < / RTI > for 5 days. After 5 days, the cells were replaced with normal medium (DMEM, containing 10% FBS) and cultured while observing until the cells morphologically changed into adipocytes.

3T3-L1 adipocytes were used to measure the triglycerol degradation promoting effect in the adipocytes of the Examples and Comparative Examples. 3T3-L1 adipocytes were washed twice with PBS (phosphate buffered saline) and then added to colorless DMEM containing 0.5% bovine serum albumin (BSA) free of fatty acids. The control group was prepared by using untreated bamboo seedlings, and the resulting value was expressed as 100% based on the medium value treated in the examples and the comparative examples. The degree of lipolysis was determined by measuring the concentration of glycerol released from adipocytes into the culture medium. Glucose was determined by a color reaction using a GPO-trinder kit purchased from Sigma Co. (St. Louis, MO, USA) and absorbance was measured at 540 nm using an ELISA reader. The results are shown in Table 6 Respectively.

division Amount of liberated glucose (%) Example 1 140 Example 2 148 Example 3 146 Example 4 162 Comparative Example 1 106 Comparative Example 2 117 Comparative Example 3 112 Comparative Example 4 106 Comparative Example 5 111 Comparative Example 6 118 Control group 100

Referring to Table 6, it can be seen that the samples of Examples 1 to 4 significantly increased the concentration of glucose liberated as compared with the samples of Comparative Examples 1 to 6, and thus the fat dissolving effect was excellent. In addition, the sample of Example 2 to which mixed extracts containing ganoderma extract, turmeric extract, and pepper seed extract were applied and the sample of Example 4 to which the mixed fermentation extract was applied had lipolytic effects compared to the single extracts of Examples 1 and 3 Further improvement.

Experimental Example  6: Cosmetics  Of the composition Slimming  Effect evaluation

Using the samples of the Examples and Comparative Examples, cream form cosmetic compositions of the same composition were prepared. Each of the above samples was formulated into cream of the same composition, but the samples of Examples 1 to 4 and Comparative Examples 1 to 6 were different.

In order to measure the slimming effect of the cream form cosmetic composition, 100 subjects (20 to 40 years old) were divided into 10 groups of 10 persons, and for each group, cream containing the samples of Examples and Comparative Examples Was applied twice a day for 1 consecutive month.

To determine the effect of slimming, we measured changes in thigh thickness before and after use of the product. The measurement method was averaged by measuring a reduced length (? L) using a ruler, and the results are shown in Table 7 below.

division Thigh circumference change (ΔL, cm) Example 1 2.8 Example 2 3.2 Example 3 3.0 Example 4 3.5 Comparative Example 1 1.4 Comparative Example 2 0.9 Comparative Example 3 1.3 Comparative Example 4 1.1 Comparative Example 5 0.8 Comparative Example 6 0.6

The results are shown in Table 7. Referring to Table 7, the cosmetic composition containing the samples of Examples 1 to 4 exhibited a significantly reduced slimming effect on the thigh area, and further, the ginseng extract, turmeric extract and pepper seed extract were further included In Example 2 and Example 4 in which the mixed fermentation extract was applied were evaluated to be remarkably excellent.

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 composition for slimming comprising a matte extract, a ginseng extract, a turmeric extract, and a black pepper seed extract as an active ingredient. delete The method according to claim 1,
Wherein the extract is a fermented extract obtained by inoculating a strain of Bifidobacterium spp . The method of claim 3,
The Bifidobacterium spp . ) Strain is Bifidobacterium bipyridinium bushes (Bifidobacterium bifidum) of the cosmetic composition. The method according to claim 1,
20 to 70 parts by weight of the matte extract, 10 to 20 parts by weight of the ginseng extract, 10 to 20 parts by weight of the turmeric extract, and 10 to 20 parts by weight of the pepper seed extract. The method according to claim 1,
Wherein the 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,
A cosmetic composition which further comprises skin moisturizing or skin soothing. 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, cleansing cream, cleansing foam, cleansing water, pack, spray, and powder.