KR20180082056A - A functional cosmetic composition comprising natural complex extract - Google Patents

Abstract

An object of the present invention is to provide a cosmetic composition which is excellent in not only functionality but also biosafety. The present invention provides a cosmetic composition comprising at least one selected from: a hydrolyzed soybean protein and Betula platyphylla sap; and a group comprising Hibiscus flower extract, Opuntia ficus flower extract, and Rosa canina extract.

Description

TECHNICAL FIELD [0001] The present invention relates to a functional cosmetic composition comprising a natural complex extract as an active ingredient,

The present invention relates to a cosmetic composition containing a natural 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.

As the industry develops, environmental pollution becomes serious, and there is a growing interest in skin aging prevention and whitening. In particular, human skin constantly undergoes changes, the most representative of which is degradation of the skin caused by aging and a reduction in visual beauty. The final appearance of skin appearance as a result of skin aging is wrinkle formation, skin elasticity decrease, senile spot formation, and wrinkle formation is considered to be the most typical phenomenon.

Lipids, proteins, polysaccharides and nucleic acids, which are major components of the skin, can be oxidized to destroy skin cells and tissues, and the skin may eventually age. Particularly, oxidation of proteins can cause collagen, hyaluronic acid, elastin, proteoglycan, fibronectin, etc., which are connective tissues of the skin, to be cleaved and hyperinflammation reaction. In the case of further deepening, mutation due to DNA mutation, Cancer induction can be promoted.

Therefore, it is necessary to protect the cell membrane by eradicating reactive oxygen species which are mediated by reactive oxygen species, ultraviolet ray irradiation, and inflammation reaction occurring during the metabolism of the body, regenerate damaged cells through active metabolism, and restore skin health desirable.

On the other hand, melanin is converted from tyrosine to dopa and dopaquinone by the action of tyrosinase existing in the pigment cells, and is produced through dopachrome and the like. Melanin protects the body from harmful factors such as ultraviolet rays and regulates hormone secretion in the body.

However, since melanin is overproduced, it can promote the formation of spots, freckles, and aging of the skin, and can be involved in the induction of skin cancer, so that research and development are actively carried out to suppress the excessive production of melanin.

(Japanese Patent Laid-open No. 4-9320), hydroquinone (Japanese Patent Laid-open No. 6-192062), kojic acid (Japanese Patent Laid-open No. 56-7710), arbutin Rosinase inhibitory activity and was used as a whitening cosmetic. However, these ingredients are poor in stability among cosmetic formulations, may be decomposed to cause coloration or odor, and their safety or efficacy in vivo is unclear and their use is limited.

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, a cosmetic product containing a natural-derived extract obtained by a conventional method has not been sufficiently functionalized, such as a skin whitening effect, and the skin improving activity is not maintained constantly.

Accordingly, there is a continuing need to develop cosmetic products derived from natural products, which have excellent functionalities and excellent biostability and skin improving 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] 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 an aspect of the present invention, there is provided a method of producing a soybean protein comprising hydrolyzed soybean protein and birch sap; And a cosmetic composition comprising at least one selected from the group consisting of Hibiscus flower extract, Paeonia sp. Flower extract, and Rosehip extract as an active ingredient.

In one embodiment, the hydrolised soy protein and birch sap may be included in an amount of 0.5-5.0 wt% based on the total weight of the composition.

In one embodiment, 10 to 30 parts by weight of the Hibiscus flower extract, 10 to 30 parts by weight of the perennial flower extract, and 10 to 30 parts by weight of the Rosehip extract may be included.

In one embodiment, the extract may be an extract obtained by fermentation inoculated with 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 cosmetic composition may be for skin whitening, skin moisturizing or skin cleansing.

In one embodiment, the cosmetic composition is comprised of a softening agent, 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, Lt; / RTI > may be formulated into one or more selected from the group.

The cosmetic composition according to one aspect of the present invention interacts with the active ingredients of the natural raw material, thus being beneficial to the skin health of the user and maximizing skin whitening and moisturizing 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 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 an aspect of the present invention, there is provided a method of producing a soybean protein comprising hydrolyzed soybean protein and birch sap; And a cosmetic composition comprising at least one selected from the group consisting of Hibiscus flower extract, Paeonia sp. Flower extract, and Rosehip extract as an active ingredient.

The cosmetic composition of the present invention contains an extract of natural origin as an active ingredient, so that it is excellent in skin improving effect and biosafety, and skin irritation can be minimized. In particular, since the mixed extract acts on the skin at the same time, skin whitening and moisturizing effect 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 above-mentioned " comprising as an active ingredient " is intended to mean an effective amount of a composition capable of exhibiting a skin improving effect, such as collagen synthesis associated with a wrinkle-reducing effect, elasticity improvement or skin whitening, skin irritation alleviation, It can mean to do.

The " hydrolyzed soy protein " means hydrolyzed protein having an average molecular weight of 2,000 to 3,000 extracted from soybean. The hydrolized soy protein promotes the synthesis of collagen by activating the skin stem cells located at the lower skin layer of the skin, and increases the elasticity of the skin. The soybean can be, but is not limited to, Seoritae, Seomotei, Blackberry, Prunus mume, Fuda bean (Chungtae), Kidney bean, Stain kidney bean, Fence bean, Mung bean, Red bean, Reed, Soybean bean, Soybean bean or Soybean.

The hydrolysis method is not particularly limited. For example, the hydrolysis may be performed by a saponification reaction by adding an acid or an alkali, and preferably by adding potassium hydroxide.

The "birch ( Betula platyphylla "is a tree belonging to birch and birch. It grows up to 20m tall, bark is white, peeled horizontally, and flowers bloom around April and May, and fruit is born in September. The birch has 7 genera and 10 plant species distributed mainly in the northern half. In Korea, 5 genera, 30 species, and 20 varieties of plants are naturally grown. As a result, birch plants have been used as a dermatological medicine by carbonizing woody parts. Is also known to have been used for nourishing gums and skin diseases.

The sap of birch tree can be used without any physical and chemical treatment of the sap collected from birch trunks, and it is possible to realize a skin improving effect such as whitening of skin including various active ingredients.

The term " Hibiscus " is a generic term for plants belonging to the genus Cercopithecidae of the dicotyledonous plants and H. rosa-sinensis in horticulture, and various hybrids produced therein. The hibiscus is known to have a diuretic effect, an antihypertensive effect and an antibacterial action because it contains a physiologically active substance of human body, and skin whitening and moisturizing effect can be realized by acting directly on the skin.

The hibiscus extract may be extracted from various organs or parts of the hibiscus (e.g., leaves, flowers, roots, stems, etc.), preferably from flowers.

The " Opuntia " Ficus Indica ) "is a group of cacti, which are native to Jeju Island, and are also called as" fan cactus "and" palm cactus ". The perennial plant has been used as a folk remedy for fever relief, bronchial asthma, dyspepsia, stomach cramps, constipation, chest pain, poor blood circulation, gastrointestinal symptoms, back pain and rhinitis. Vitamin C is 5 times more than aloe And calcium and dietary fiber.

The above-mentioned " Rosa canina " refers to the fruit after the blooming of the blooming flowers and is also called "Rose Haw". The rose hips are native to Central Asia and Turkey (YamanKaradeniz, 1983) and have been used in tea and vitamin C supplements and food medicines for years in Europe and the United States. According to the Herb Pharmacopoeia in the UK, Rosehip is said to be effective against gastritis, diarrhea, thirst and vitamin C deficiency. It contains 20 to 40% more vitamin C than vegetables and fruits. Can be implemented.

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 then pulverizing the nanocloprosis oseanica, 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) of ethanol, and more particularly, it can be extracted with a concentration of 60 to 80% (v / v) 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

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 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 >

The hydrolized soybean protein and birch sap may be contained in an amount of 0.5 to 5.0 wt% based on the total weight of the composition. If the content of the hydrolyzed soybean protein and the birch sap is less than 0.5% by weight, the effect of improving the skin may not be properly realized. If the content is more than 5.0% by weight, the proportion of other ingredients required for commercialization is decreased, The merchantability may be lowered and the cost efficiency may be lowered.

10 to 30 parts by weight of the hibiscus flower extract, 10 to 30 parts by weight of the perennial flower extract, and 10 to 30 parts by weight of the rosehip 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 inoculated with 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 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 extract or the resin extract to a certain concentration in a solvent, preferably purified water, For example, by inoculating and fermenting a microorganism, that is, a Lactobacillus sp . Strain.

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.

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 , And the alcohol may be ethanol at a concentration of 60 to 90% (v / v).

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

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 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 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. 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: Manufacture of flower extracts

Hibiscus flower, Paekcheoncho flower and Rosehip were washed with water, completely dried at room temperature and pulverized to obtain 100 g of pulverized product, respectively.

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 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 fermented extract was filtered with a 500 mesh and a 0.2 mu m filter to obtain a homogeneous fermented extract.

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. Hydrolyzed soy protein and birch sap were obtained commercially and used.

The skin improvement effect was measured by mixing the respective raw materials at a predetermined ratio as shown in Table 1 below. In Example 8, the fermented extract of Production Example 2 was mixed.

[Content (parts by weight)] division Hydrolyzed
Soy protein Birch sap Hibiscus flower extract Flower extract Rosehip extract Example 1 20 20 30 - - Example 2 20 20 - 30 - Example 3 20 20 - - 30 Example 4 20 20 15 15 - Example 5 20 20 - 15 15 Example 6 20 20 15 - 15 Example 7 20 20 10 10 10 Example 8 20 20 10 (Preparation Example 2) 10 (Preparation Example 2) 10 (Preparation Example 2) Comparative Example 1 35 35 - - - Comparative Example 2 - - 70 Comparative Example 3 - - 70 Comparative Example 4 - - 70 Comparative Example 5 - - 25 25 25

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 sample is not only harmless to the human body but also has excellent human safety.

Experimental Example  2: Evaluation of melanin production

Melanin contents assays were used to evaluate the inhibition of melanogenesis by the samples of the Examples and Comparative Examples (Eisinger M et al, 1982; Siegrist W and Eberle AN, 1986). The samples of the examples and comparative examples were each suspended in purified water at a concentration of 50 mg / L and the amount of melanin produced was evaluated.

B16F10 cells were inoculated in a 60 mm culture dish at a concentration of ^1.5 × 10 ⁵ cells / well and cultured for 24 hours. Α-MSH (Sigma-Aldrich, St. Louis, MO, USA), a hormone that promotes melanin formation, and samples of the examples and comparative examples were added to the medium at a concentration of 50 mg / L (Virador VM et al., 1999).

After 48 hours, the cells were harvested and washed once with PBS. Then, 120 μL of 1 N NaOH lysis buffer was added and the cells were lysed by heating at 95 ° C. for 10 minutes. Absorbance was measured at 450 nm wavelength using a microplate reader. Arbutin (100 μg / mL) was used as a control.

The inhibition rate of melanin production was calculated according to the following equation, and the evaluation results are shown in Table 2 below.

[Mathematical Expression]

Melanin production inhibition rate (%) = 100 x (1 - (absorbance of each test substance / absorbance of control))

division Melanin production inhibition rate (%) Example 1 26.7 Example 2 29.2 Example 3 27.7 Example 4 33.2 Example 5 31.7 Example 6 32.5 Example 7 37.2 Example 8 43.1 Comparative Example 1 18.5 Comparative Example 2 13.5 Comparative Example 3 14.2 Comparative Example 4 11.6 Comparative Example 5 17.6 Positive control group 32.4

The samples of Examples 1 to 8 effectively inhibited the production of melanin and inhibited the production of melanin to an equivalent or higher level compared with albutin, the whitening effect being widely known.

In particular, when the extracts of at least one of Hibiscus flower extract, Paeonia spp. Flower extract and Rose Hip extract acted together, the melanin formation inhibitory effect was further improved, and the whitening effect was remarkably increased by fermentation of the extract (Example 8).

Experimental Example  3: tyrosinase inhibitory activity test

The tyrosinase inhibitory activity was measured in vitro to compare the whitening effects of the examples and the comparative examples.

Each of the samples of the examples and comparative examples was suspended in purified water at a concentration of 50 mg / L, and then 220 μL of 0.1 M phosphate buffer (pH 6.5), 20 μL of the sample solution, tyrosinase (1500 U / mL to 2000 U / 20 μL was added sequentially. 40 μl of a 1.5 mM tyrosinase solution was added thereto and reacted at 37 ° C for about 15 minutes.

Absorbance was measured at 490 nm using an ELISA reader. A 0.1 M phosphate buffer (pH 6.5) was used instead of the sample solution.

As a control, kojic acid at a concentration of 50 μM was used, and tyrosinase inhibitory activity was calculated according to the following equation. The evaluation results are shown in Table 3 below.

[Mathematical Expression]

Tyrosinase inhibitory activity (%) = 100 x (1 - (absorbance of each test substance / absorbance of negative control))

division Tyrosinase inhibitory activity (%) Example 1 25.1 Example 2 30.2 Example 3 27.7 Example 4 36.2 Example 5 35.7 Example 6 33.9 Example 7 38.2 Example 8 43.6 Comparative Example 1 12.2 Comparative Example 2 11.5 Comparative Example 3 10.9 Comparative Example 4 9.6 Comparative Example 5 17.5 Positive control group 29.4

The samples of Examples 1 to 8 inhibited the activity of tyrosinase, and the tyrosinase inhibitory activity was evaluated to be higher or equal to that of the positive control (kojic acid).

In particular, when the extracts of Hibiscus flower extract, Paeonia sp. Flower extract, and at least one extract of Rose Hip extract acted together, tyrosinase inhibitory activity was further improved, and the extract had enhanced tyrosinase inhibitory activity by fermentation (see Example 8).

The results show that the effective components of the flower extract interact with the hydrolized soy protein and birch sap, thereby significantly improving the whitening effect.

Experimental Example  4: 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 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.

[% of control] division Collagen production (%) Example 1 121.7 Example 2 123.5 Example 3 119.7 Example 4 127.4 Example 5 126.8 Example 6 129.5 Example 7 135.4 Example 8 140.8 Comparative Example 1 114.5 Comparative Example 2 110.2 Comparative Example 3 109.7 Comparative Example 4 111.2 Comparative Example 5 113.1 Positive control group 133.2

Experimental Example  5: skin moisturizing effect test

Samples of the examples and comparative samples 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 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.

These results suggest that the hydrolized soy protein and birch sap interact with the active ingredient of the extract to effectively promote the expression of the aquaporin protein and the synthesis of hyaluronic acid, thereby improving the skin moisturizing effect do.

[% of control] division HAS2 expression level AQP3 expression level Example 1 121.5 119.8 Example 2 123.2 122.3 Example 3 125.6 121.1 Example 4 133.5 129.8 Example 5 136.8 134.5 Example 6 135.3 132.3 Example 7 141.9 139.8 Example 8 145.5 144.2 Comparative Example 1 113.9 109.4 Comparative Example 2 108.1 106.5 Comparative Example 3 111.5 109.2 Comparative Example 4 112.2 109.9 Comparative Example 5 114.7 111.4

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)

Hydrolyzed soy protein and birch sap; And
A hibiscus flower extract, a papaya flower extract, and a rose hip extract as an active ingredient. The method according to claim 1,
Wherein the hydrolized soy protein and birch sap are 0.5 to 5.0 wt% based on the total weight of the composition. The method according to claim 1,
10 to 30 parts by weight of the Hibiscus flower extract, 10 to 30 parts by weight of the flower extract of Paeonia sp., And 10 to 30 parts by weight of the rosehip extract. The method according to claim 1,
The extract is an extract obtained by fermentation inoculated with the Lactobacillus genus (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. 7. The method according to any one of claims 1 to 6,
Skin whitening, skin moisturizing or skin soothing. 7. The method according to any one of claims 1 to 6,
Cosmetics formulated with 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 Composition.