KR102628844B1 - Cosmetic composition for anti-oxidation, skin whitening and anti-wrinkle containing a mixed fermentation extracts of Wisteria floribunda flowers, Aralia elata flowers, Camellia japonica leafs and Camellia japonica flowers as effective component - Google Patents

Abstract

The present invention relates to a cosmetic composition for skin whitening and improving skin wrinkles containing wisteria flowers, aralia flowers, camellia leaves and mixed fermented extracts of camellia flowers.
The cosmetic composition for antioxidant, skin whitening, and skin wrinkle improvement according to the present invention is composed of natural substances, is safe for the skin, and has excellent skin whitening and skin wrinkle improvement effects. More specifically, the composition of the present invention effectively acts on skin whitening by inhibiting melanin synthesis and tyrosinase activity, effectively inhibits the expression of MMP-1, which directly acts on the creation of skin wrinkles, and increases the expression of collagen protein. It works effectively in preventing and improving skin wrinkles.

Description

Cosmetic composition for anti-oxidation, skin whitening and anti-wrinkle improvement containing wisteria flowers, aralia flowers, camellia leaves and mixed fermented extracts of camellia flowers as active ingredients {Cosmetic composition for anti-oxidation, skin whitening and anti-wrinkle containing a mixed fermentation extracts of Wisteria floribunda flowers, Aralia elata flowers, Camellia japonica leafs and Camellia japonica flowers as effective component}

The present invention relates to a cosmetic composition for skin whitening and improving skin wrinkles containing wisteria flowers, aralia flowers, camellia leaves and mixed fermented extracts of camellia flowers.

Recently, interest in cosmetics derived from natural products is growing. In addition, as the demand for functional cosmetics increases, the problem of the skin's protective ability and regenerative ability being reduced due to various chemical ingredients used as functional ingredients is occurring. Accordingly, it is becoming popular to make and use cosmetics made from natural materials, ranging from toners and lotions to color cosmetics. In the case of people with skin diseases such as atopy or sensitive skin, interest in cosmetics derived from natural products that minimize skin irritation and have excellent skin improving activity is increasing.

The present inventors conducted research on natural materials that are non-toxic and have excellent antioxidant, skin whitening, and skin wrinkle improvement effects, and discovered that fermented wisteria flower extract exhibits excellent antioxidant, skin whitening, and skin wrinkle improvement effects. In addition, while continuing research on the fermented wisteria flower extract, when wisteria flowers were fermented by mixing them with aralia flowers, camellia leaves, and camellia flowers, not only did the skin whitening and skin wrinkle improvement effects improve, but they also had excellent antioxidant effects. The present invention was completed by discovering that .

KR 10-2011-0001538 A KR 10-2018-0005991 A

The problem to be solved by the present invention is to provide a cosmetic composition for antioxidant, skin whitening or skin wrinkle improvement containing wisteria flowers, aralia flowers, camellia leaves and mixed fermented extracts of camellia flowers as active ingredients.

Another object of the present invention is to provide a food composition for antioxidant, skin whitening or skin wrinkle improvement containing the above-mentioned wisteria flowers, aralia flowers, camellia leaves and mixed fermented extracts of camellia flowers as active ingredients.

In order to achieve the above object, the present invention provides a cosmetic composition for antioxidant, skin whitening, or skin wrinkle improvement containing fermented extracts of wisteria flowers, aralia flowers, camellia leaves, and camellia flower mixtures as active ingredients.

According to one embodiment of the present invention, the fermentation extract of the mixture may be included in an amount of 0.0001 to 20% by weight based on the total weight of the cosmetic composition.

According to one embodiment of the present invention, the mixture may include wisteria flowers, aralia flowers, camellia leaves, and camellia flowers in a weight ratio of 1:0.05-0.5:0.2-2:0.1-1.

According to one embodiment of the present invention, the mixture may further include 10 to 20 parts by weight of Ubangja based on 100 parts by weight of the mixture.

According to one embodiment of the present invention, the fermentation may be performed using Lactobacillus rhamnosus bacteria.

According to one embodiment of the present invention, the fermented extract of the mixture may be prepared by fermenting the mixture and then performing ultra-high pressure extraction.

According to one embodiment of the present invention, the fermented extract of the mixture may be a cosmetic composition characterized by exhibiting the following activities (1) to (5):

(1) activity to suppress expression of MMP-1 protein;

(2) activity to promote the expression of procollagen type I protein;

(3) tyrosinase activity inhibitory activity;

(4) melamine synthesis inhibitory activity; and

(5) DPPH (1-1-diphenyl-2-picryl-hydrazyl) radical scavenging activity.

According to one embodiment of the present invention, the cosmetic composition may be a formulation selected from lotions, emulsions, creams, essences, cosmetic ointments, sprays, gels, packs, sunscreens, makeup bases, foundations, powders, and makeup removers.

In addition, the present invention provides a food composition for antioxidant, skin whitening, or skin wrinkle improvement comprising fermented extracts of wisteria flowers, aralia flowers, camellia leaves, and camellia flower mixtures as active ingredients.

The cosmetic composition for antioxidant, skin whitening and skin wrinkle improvement according to the present invention contains natural substances such as wisteria flower, aralia flower, camellia leaf and fermented extract of camellia flower mixture as active ingredients, so it is safe for the skin and has excellent skin properties. It has whitening and skin wrinkle improvement effects. More specifically, the composition of the present invention effectively acts on skin whitening by inhibiting melanin synthesis and tyrosinase activity, effectively inhibits the expression of MMP-1, which directly acts on the creation of skin wrinkles, and increases the expression of collagen protein. It works effectively in preventing and improving skin wrinkles.

Hereinafter, the present invention will be described in detail.

According to one embodiment of the present invention, the present invention provides a cosmetic composition for antioxidant, skin whitening or skin wrinkle improvement comprising fermented extract of wisteria flower, aralia flower, camellia leaf and camellia flower mixture as an active ingredient. .

The present inventors have made extensive research efforts to increase the usability of natural plant materials, and as a result, when fermenting a mixture of wisteria flowers, aralia flowers, camellia leaves, and camellia flowers, it is effective in skin whitening and improving skin wrinkles. It was found to have excellent effects, and it was also confirmed that there were no safety issues when applied to the skin.

The composition of the present invention is non-cytotoxic, has excellent antioxidant effect, and inhibits matrix protease-1 (matrix metallo-proteinase, hereinafter referred to as 'MMP') protein expression and promotes procollagen type I protein expression, thereby protecting the skin. It has the characteristic of effectively suppressing and improving wrinkles. Therefore, the fermented extract of the wisteria flower, aralia flower, camellia leaf and camellia flower mixture of the present invention significantly improves skin whitening and skin wrinkle improvement by using natural plant materials that are easily available on the market and have guaranteed safety. It has the advantage of being effective.

In this specification, the “fermentation extract of the mixture” may be described as “mixed fermentation extract.”

Additionally, in this specification, the term “wisteria flower mixed fermented extract” means “fermented extract of wisteria flower, aralia flower, camellia leaf and camellia flower mixture.”

In the present invention, “ Wisteria floribunda ” is a tree of the legume family. It is a deciduous, vine-leafed tree whose vines grow over 10m long and turn to the right, wrapping around other objects. The leaves are alternate, pinnate compound leaves, and have 4 to 6 pairs of small leaves, which are egg-shaped with pointed ends and have short petioles. In spring, many blue-purple butterfly flowers grow in racemes tens of centimeters long from the leaf axils. The fruit forms a long capsule about 15cm long, which hangs down and opens when ripe to pop out the seeds. In the present invention, wisteria flowers were used.

In the present invention, “ Aralia elata ” is a deciduous broad-leaved shrub of the Aralia family. The shoots are edible, and the whole bark of herbal medicine is dried tree bark. In oriental medicine, the fruits and roots are used in seawater, stomach cancer, diabetes, and digestive medicine. It is used in the private sector to treat diabetes by decocting the bark or roots. In the present invention, aralia tree flowers were used.

In the present invention, “ Camellia japonica L. ” belongs to the Camellia family and is an evergreen broad-leaved small tree that grows in Gochang, Jeollabuk-do, Haenam, Jeollanam-do, Wando, Gangjin, Yeosu, Gwangyang, Gyeongnam, Geoje, the southern coastal region, and Jeju Island. It grows to about 15 m tall and about 50 cm in diameter. The leaf surface is dark green and glossy, the back side is yellow-green, oval-shaped, 5-12 cm long and 3-7 cm wide, with wavy fine teeth. There is. In the present invention, camellia leaves and flowers were used.

In the present invention, “Woobangja” is the fruit of burdock ( Arctium lappa L. ), a biennial herb of the Asteraceae family, and is also known by other names such as Daedoja, Woochaeja, and Black Fengja. The achenes grow in September and ripen to grayish brown, with brown caps attached and black seeds coming out from the inside. The main effect of Ubangja is that it is effective in relieving symptoms such as seawater, sputum, and sore throat caused by wind heat.

The mixed fermented extract of wisteria flowers, aralia flowers, camellia leaves and camellia flowers used as active ingredients in the composition of the present invention is a mixture of extracts extracted from each of wisteria flowers, aralia flowers, camellia leaves and camellia flowers. It refers to an extract of a post-fermented fermented product, or an extract of a fermented product obtained by mixing wisteria flowers, aralia flowers, camellia leaves, and camellia flowers.

In the present invention, the mixture may include wisteria flowers, aralia flowers, camellia leaves and camellia flowers at a weight ratio of 1:0.05-0.5:0.2-2:0.1-1, preferably 1:0.1-0.4:0.5. -A weight ratio of 1.5:0.25-0.75, more preferably a weight ratio of 1:0.15-0.35:0.8-1.2:0.4-0.6, more preferably a weight ratio of 1:0.2-0.3:0.9-1.1:0.45-0.55, most preferably may be a weight ratio of 1:0.25:1:0.5. When each weight ratio is within the above range, the synergistic effect of the mixed fermentation of the wisteria flowers, aralia flowers, camellia leaves, and camellia flowers can be maximized. On the other hand, if the weight ratio of the mixture is outside the above range, the skin whitening and skin wrinkle improvement efficacy will not meet expectations.

In particular, in the present invention, the mixture may include the camellia leaves and camellia flowers at a weight ratio of 2:0.5-1.5, preferably at a weight ratio of 2:0.8-1.2, and more preferably at a weight ratio of 2:1. there is.

In addition, in a preferred embodiment, the mixture may further include 10 to 30 parts by weight, preferably 10 to 20 parts by weight, and more preferably 12 to 18 parts by weight, based on 100 parts by weight of the mixture. When the above-mentioned algae is added in the weight ratio relative to the mixture, the antioxidant, skin whitening, and skin wrinkle improvement effects are further improved. If the content of algae in the mixture is less than the lower limit, the effect due to the addition of algae becomes insignificant, and if it exceeds the upper limit, the antioxidant, skin whitening, and skin wrinkle improvement effects increased by the addition of algae are rather reduced. There is a problem.

In addition, the mixed fermented extract of the present invention may be prepared by mixing the mixture with distilled water according to a conventional method and then fermented, or it may be prepared as a mixed extract and then fermented. In addition, the mixed extract is not particularly limited, but may be extracted using a solvent extraction method or an ultra-high pressure treatment extraction method.

The mixed extract obtained by solvent extraction may be extracted according to a common method known in the art. For example, the mixed extract by solvent extraction is a mixture of wisteria flowers, aralia flowers, camellia leaves and camellia flowers with the extraction solvent at a ratio of 1:10 to 30, preferably 1:15 to 25, more preferably. is mixed at a weight ratio of 1:18 to 22, extracted at 65 to 95°C, preferably 70 to 90°C, more preferably 75 to 85°C for 1 to 10 hours, preferably 1 to 5 hours, and then concentrated under reduced pressure. It may be manufactured by performing . If the weight ratio is outside the above range, the active ingredients of wisteria flowers, aralia flowers, camellia leaves, and camellia flowers may be extracted in small amounts.

The extraction solvent for the solvent extraction may be water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. The lower alcohol may include 20 to 80% methanol, ethanol, butanol, or propanol.

The extraction solvent is not particularly limited, but an extract extracted with a 60 to 80% ethanol aqueous solution is preferred for skin whitening and improving skin wrinkles.

In addition, the mixed extract by ultra-high pressure extraction may be extracted by ultra-high pressure treatment of a mixture of wisteria flowers, aralia flowers, camellia leaves, and camellia flowers. At this time, ultra-high pressure treatment means treatment at a pressure of 100 to 3000 MPa and a temperature of 40 to 90°C for 1 to 24 hours, preferably at a pressure of 500 to 1000 MPa at a temperature of 40 to 60°C and 12 to 24°C. It is a time treatment, and most preferably means treatment at a pressure of 1000 MPa at a temperature of 50° C. and for 24 hours. For the ultra-high pressure treatment, water, anhydrous alcohol having 1 to 4 carbon atoms, ethyl acetate, acetone, glycerin, ethylene glycol, propylene glycol, butylene glycol, caprylic/capric triglyceride, dimethicone, mineral oil, and cyclomethicone. At least one extraction solvent selected from the group consisting of octyldodecanol, cetyl ethylhexanoate, triethylhexanoin, isopropyl myristate, and vegetable oil can be added.

The suspension or mixed extract of the mixture obtained as described above can be prepared as a mixed fermented extract by fermenting the mixture for 1 to 7 days using fermentation bacteria while maintaining pH 5 to 7 under temperature conditions of 20 to 40°C.

At this time, the suspension or mixed extract is preferably added in an amount of 1 to 20% by weight to the basic medium of fermenting bacteria.

The fermenting bacteria may be yeast, lactic acid bacteria, Bacillus genus bacteria, and mixtures thereof, preferably lactic acid bacteria, more preferably Lactobacillus genus lactic acid bacteria, and even more preferably Lactobacillus rhamnonus and Lactobacillus planta. It may be one or more types of lactic acid bacteria selected from Rum, and Lactobacillus acidophilus, more preferably Lactobacillus rhamnonus, and most preferably Lactobacillus rhamnonus HK-9 (KCCM11254P4).

The fermentation bacteria may be added as the fermentation bacteria themselves or may be inoculated into the culture medium. The culture medium may include the culture medium itself obtained by culturing the strain in a liquid medium, a concentrate obtained by concentrating the culture medium, etc.

In one embodiment of the present invention, the mixed fermentation extract is a suspension of a mixture of wisteria flowers, aralia flowers, camellia leaves and camellia flowers, or a mixture of Lactobacillus rhamnonus HK-9 (KCCM11254P4) in an appropriate amount, e.g. For example, it may be obtained by inoculating at a concentration ^{of 1} there is.

In addition, the mixed fermented extract of the present invention may be obtained by fermenting the mixture and then extracting it under ultra-high pressure to increase skin whitening and skin wrinkle improvement activity.

In a preferred embodiment, the mixed fermented wisteria flower extract of the present invention includes the steps of solvent extracting a mixture of wisteria flowers, aralia flowers, camellia leaves, and camellia flowers to obtain a wisteria flower mixed extract; Obtaining a mixed fermented wisteria flower extract by fermenting the wisteria flower mixed extract with bacteria selected from yeast, lactic acid bacteria, Bacillus bacteria, and mixtures thereof; And the step of ultra-high pressure extraction of the wisteria flower mixed fermentation extract at a pressure of 100 to 3000 MPa and a temperature of 40 to 90° C. for 1 to 24 hours. In this way, when an ultra-high pressure extraction step is added to the mixed fermented wisteria flower extract of the present invention, the skin whitening and skin wrinkle improvement efficacy is significantly increased compared to the case where ultra-high pressure extraction is not performed.

In addition, the mixed fermentation extract of the present invention may be subjected to additional processes such as filtration, organic solvent reflux extraction, reduced pressure concentration, and freeze-drying after fermentation. It is obvious in the art that the mixed fermentation extract obtained by filtering, purifying and concentrating the above-described mixed fermentation extract is also included in the present invention.

In particular, in one embodiment of the present invention, the mixed fermented extract of wisteria flowers has superior MMP-1 protein expression inhibition activity and procollagen compared to the single fermented extract of each plant due to the synergistic effect or collaboration between the active ingredients in the fermented extract. It was confirmed that it has the activity of promoting the expression of type I protein, the activity of inhibiting tyrosinase activity, and the activity of inhibiting melamine synthesis.

Specifically, the mixed fermented extract of wisteria flowers of the present invention reduces the expression of MMP-1 protein more than the single fermented extract of wisteria flowers, aralia flowers, camellia leaves, and camellia flowers, and increases the expression of procollagen type I protein. was further increased, resulting in greater skin wrinkle improvement efficacy.

In particular, the mixed fermented extract of wisteria flowers of the present invention inhibits tyrosinase activity and melamine synthesis more than the single fermented extract of each plant due to a synergistic or collaborative effect between the active ingredients in the mixed fermented extract, resulting in better skin. It was able to show whitening efficacy.

Meanwhile, the inventors of the present invention tested the cumulative skin irritation of the composition through specific experiments and confirmed that it was a harmless substance to the human body. In addition, the mixed fermented extract of wisteria flowers of the present invention has almost no toxicity and side effects, so it can be used safely even for long-term use.

In addition, the mixed fermentation extract of the present invention also includes processed products of the mixed fermentation extract. For example, the mixed fermented extract of wisteria flowers of the present invention can be produced in powder form by additional processes such as distillation under reduced pressure and freeze-drying or spray-drying after fermentation extraction.

In addition, the mixed fermentation extract of the present invention, in a broad sense, is a processed product of a mixed fermentation extract of wisteria flowers, aralia flowers, camellia leaves and camellia flowers formulated to be applied or administered to animals, such as wisteria flowers. It is also meant to include mixed fermentation extract powder. Although experiments were conducted with a mixed fermented wisteria flower extract in the present invention, those skilled in the art would be able to predict that the desired effect can be achieved even in the same form as a processed wisteria flower mixed fermented extract.

In the present invention, “containing as an active ingredient” means containing an amount sufficient to achieve the efficacy or activity of the mixed fermented extract of wisteria flowers.

In an exemplary embodiment, the wisteria flower mixed fermentation extract may be present in the composition in any amount desired to provide effective antioxidant, skin whitening, and skin wrinkle improvement effects. According to one aspect, the mixed fermentation extract may be 0.0001 to 99.9% by weight, preferably 0.0001 to 30% by weight, more preferably 0.0001 to 20% by weight, and even more preferably 0.001 to 0.001% by weight, based on the total weight of the cosmetic composition. It may be included at 20% by weight, more preferably at 0.001 to 10% by weight, and even more preferably at 0.005 to 5% by weight. If the content of the wisteria flower mixed fermentation extract is less than the lower limit, no skin improvement effect appears, and if it exceeds the upper limit, the degree of increase in effect due to increase in content is minimal, there are problems with the safety and stability of the formulation, and it is economical. It is also undesirable from a human perspective. In other words, by including the mixed fermentation extract of the present invention in the above range, it is possible to achieve excellent antioxidant, skin whitening and skin wrinkle improvement effects, has formulation and product stability, and can exhibit excellent effects at the optimal content in terms of economic efficiency. . As a result of an in vitro experiment in one embodiment of the present invention, when the concentration of the mixed fermented extract of wisteria flowers of the present invention is within the above range, significant effects on antioxidant, skin whitening, and skin wrinkle improvement are observed, while side effects such as cytotoxicity are observed. didn't show up

The cosmetic composition of the present invention can exhibit a whitening effect by containing the wisteria flower mixed fermentation extract as an active ingredient, and more specifically, it contains the whitening active ingredient derived from the wisteria flower mixed fermentation extract as an active ingredient, resulting in significant tyrosinase. It has an activity-inhibiting effect and can exhibit an inhibitory effect on melanin biosynthesis.

In one embodiment of the present invention, the skin wrinkles may be caused by photoaging or natural aging, more preferably photoaging, and more preferably photoaging due to UV exposure, but are not limited thereto. The skin wrinkles are preferably based on the activity of MMP enzymes, but are not limited thereto. In general, the formation of skin wrinkles is promoted due to increased expression and activity of MMP, an enzyme that decomposes collagen, a major component of the skin, after UV exposure.

In addition, the cosmetic composition containing the mixed fermented extract of wisteria flowers as an active ingredient is characterized by increasing the expression of type I procollagen protein and suppressing the expression of MMP-1 protein.

Specifically, the cosmetic composition containing the mixed fermented extract of wisteria flowers of the present invention inhibits the expression of matrix metallo-proteinase, which promotes collagen decomposition, while increasing the production of type I procollagen. Bar, increasing skin elasticity, skin regeneration, improving skin wrinkles, wound healing, repair and regeneration of damaged skin tissue; and may have effects such as preventing skin aging. In other words, it can have the effect of preventing or improving skin wrinkles.

The mixed fermented extract of wisteria flowers included in the cosmetic composition for improving skin wrinkles of the present invention has the effect of removing skin wrinkles by itself, and when used by mixing with substances known to be effective in improving skin wrinkles. The effect of preventing or improving skin wrinkles can be maintained continuously.

According to a preferred embodiment of the present invention, the cosmetic composition for antioxidant, skin whitening, and skin wrinkle improvement may additionally include a dermatologically acceptable excipient in addition to the wisteria flower mixed fermentation extract.

The excipients are not limited thereto, but may include, for example, skin emollients, skin penetration enhancers, colorants, fragrances, emulsifiers, thickeners, and solvents. In addition, it may additionally contain fragrances, pigments, disinfectants, antioxidants, preservatives, and moisturizers, and may include thickeners, inorganic salts, synthetic polymer materials, etc. for the purpose of improving physical properties.

According to a preferred embodiment of the present invention, the moisturizing agent is glycerin, butylene glycol, propylene glycol, sorbitol, hexylene glycol, dipropylene glycol, diglycerin, 1,2-hexanediol, panthenol, betaine, squalane, petrolatum, liquid. It may be one or more selected from paraffin and hydrolyzed wheat gluten, but is not limited thereto.

The cosmetic composition of the present invention can be prepared in any formulation commonly prepared in the art, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing agents. , oil, powder foundation, emulsion foundation, wax foundation, spray, etc., but is not limited thereto. More specifically, it can be manufactured in the form of flexible lotion (skin), nourishing lotion (milk lotion), nourishing cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray, or powder. .

When the cosmetic composition of the present invention is a soap, a surfactant-containing cleansing formulation, or a surfactant-free cleansing formulation, it can be applied to the skin and then wiped off, removed, or washed with water. As a specific example, the soap is liquid soap, powdered soap, solid soap, and oil soap, the surfactant-containing cleansing formulation is cleansing foam, cleansing water, cleansing towel, and cleansing pack, and the surfactant-free cleansing formulation is cleansing cream. , cleansing lotion, cleansing water, and cleansing gel, but are not limited thereto.

The cosmetic composition of the present invention can be used alone or in multiple applications, or can be used in multiple applications with other cosmetic compositions other than the present invention. Additionally, the cosmetic composition according to the present invention can be used according to conventional usage methods, and the number of times of use can be varied depending on the user's skin condition or preference.

According to another aspect of the present invention, the present invention relates to a skin health functional food composition for antioxidant, skin whitening and skin wrinkle improvement comprising wisteria flowers, aralia flowers, camellia leaves and mixed fermented extracts of camellia flowers as active ingredients. will be. The term “mixed fermentation extract” in the present invention is as described above.

The health functional food composition of the present invention can be useful for antioxidant, skin whitening, and skin wrinkle improvement.

The term “prevention” in the present invention means anything that suppresses or delays the increase in skin pigmentation or skin wrinkles.

The term “improvement” in the present invention means at least reducing the degree of parameters related to the condition being treated, such as skin pigmentation or skin wrinkling, by administration of a composition comprising the mixed fermented extract of wisteria flowers of the present invention.

The health functional food composition may be provided in the form of powder, granules, tablets, capsules, syrup, or beverage, and the health functional food composition may be used with other foods or food additives in addition to the active ingredients, and may be administered appropriately according to conventional methods. It can be used effectively. The mixing amount of the active ingredient can be appropriately determined depending on its purpose of use, for example, prevention, improvement or therapeutic treatment. According to one aspect, the wisteria flower mixed fermentation extract is 0.001 to 30% by weight, preferably 0.001 to 20% by weight, more preferably 0.001 to 10% by weight, more preferably 0.01 to 10% by weight, even more preferably May be included in 0.1 to 5% by weight.

The effective dose of the active ingredient contained in the health food composition is the type and content of the active ingredient and other ingredients contained in the composition, type of dosage form, age, weight, general health condition, gender and diet of the ingestor, administration time, and administration. It may be adjusted depending on various factors including route, period of intake, and concurrently used drugs, but is not limited thereto. In addition, in the case of long-term intake for the purpose of skin whitening and skin wrinkle improvement efficacy, the amount may be below the above range, and since the active ingredient has no safety issues, it is certain that it can be used in amounts above the above range.

Hereinafter, the present invention will be described in detail through examples, but the present invention is not limited to the following examples.

< Examples and Comparative Examples>

Example 1: Preparation of wisteria flower mixed fermentation extract - weight ratio of 1:0.25:1:0.5

(1) Preparation of wisteria flower mixed extract

Wisteria flowers, aralia flowers, camellia leaves, and camellia flowers were mixed at a weight ratio of 1:0.25:1:0.5, pulverized, and passed through a 300 mesh sieve to prepare the pulverized product.

The mixed pulverized product and 70% ethanol aqueous solution were mixed at a weight ratio of 1:20 and extracted at 80° C. for 3 hours to obtain a wisteria flower mixed solvent extract. The obtained extract was filtered, concentrated under reduced pressure, and powdered by freeze-drying.

(2) Preparation of wisteria flower mixed fermentation extract

1) Lactobacillus rhamnosus HK-9 (accession number: KCCM11254P) obtained from the Korea Microorganism Conservation Center was placed in a shaking constant-temperature incubator with a medium prepared by adding 55% MRS medium and 0.5% L-Cysteine hydrochloride, and incubated for 50 minutes. A lactic acid bacteria culture medium was prepared by culturing at ~60 rpm and 37°C for 48 hours.

2) 5% by weight of the wisteria flower mixed extract obtained in step (1) was mixed with distilled water and sterilized at 121°C for 15 minutes to prepare a wisteria flower mixed extract for fermentation.

3) 5% (v/v) of lactic acid bacteria culture medium (3×10 ⁹ cfu/ml) was inoculated into the wisteria flower mixed extract, and cultured with shaking at 37°C and 150 rpm for 48 hours. Afterwards, the fermented product was centrifuged at 3,000 rpm for 10 minutes to obtain the supernatant, filtered using 0.25 μM filter paper, concentrated under reduced pressure, and powdered by freeze-drying.

Example 2: Preparation of wisteria flower mixed fermentation extract - weight ratio of 1:1:1:1

The same procedure as in Example 1 was carried out, except that wisteria flowers, aralia flowers, camellia leaves, and camellia flowers were mixed at a weight ratio of 1:1:1:1 to prepare a mixed fermented extract of wisteria flowers.

Example 3: Preparation of Wisteria flower mixed fermentation extract - Addition of Wisteria flower

The same procedure as in Example 1 was performed, except that 15 parts by weight of Wisteria flowers were further mixed with 100 parts by weight of the mixture of wisteria flowers, aralia flowers, camellia leaves, and camellia flowers to prepare a mixed fermented extract of wisteria flowers.

Example 4: Preparation of wisteria flower mixed ultra-high pressure extract

After adding twice the volume of distilled water to the mixed fermented extract of wisteria flowers of Example 1, the temperature was raised to 50°C under a pressure of 1000 MPa using a high pressure processor, ultra-high pressure treated for 24 hours, and then cooled to room temperature. The ultra-high pressure treated Wisteria flower mixed fermentation extract and 70% (v/v) ethanol aqueous solution were mixed at a weight ratio of 1:20 and extracted at 80°C for 3 hours to obtain the Wisteria flower mixed fermentation ultra-high pressure extract. The obtained extract was filtered, concentrated under reduced pressure, and powdered by freeze-drying.

Comparative Example 1: Wisteria flower sole fermented extract

The same procedure as Example 1 was carried out, except that only wisteria flowers were used to prepare a fermented extract of wisteria flowers alone.

Comparative Example 2: Fermented extract of aralia tree flowers alone

The same procedure as in Example 1 was carried out, except that only the flowers of the aralia tree were used to prepare a fermented extract of the aralia flower alone.

Comparative Example 3: Camellia leaf fermented extract alone

The same procedure as in Example 1 was performed, but a fermented extract of camellia leaves alone was prepared using only camellia leaves.

Comparative Example 4: Camellia flower isolated fermented extract

The same procedure as Example 1 was carried out, except that only camellia flowers were used to prepare a fermented extract of camellia flowers alone.

Comparative Example 5: Mixed fermentation extract excluding wisteria flowers

A mixed fermentation extract was prepared in the same manner as in Example 1, except that wisteria flowers were excluded and only aralia flowers, camellia leaves, and camellia flowers were used.

Comparative Example 6: Wisteria flower mixed extract - fermentation omitted

A mixed extract of wisteria flowers was prepared in the same manner as in Example 1, but only step (1) was performed.

< Test example>

statistical processing

All statistical analyzes were performed using SPSS version 18.0 (IBM, Chicago, IL, USA). Differences in mean values between experimental groups (p <0.05) were determined using one-way ANOVA with Duncan's post-hoc test. The following experiments were each repeated at least three times, and each experimental data was expressed as mean ± standard deviation (SD).

Test Example 1: Cytotoxicity evaluation using MTT assay

Cytotoxicity was measured using the MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) reduction method.

The pre-cultured HaCaT cells were mixed well in DMEM medium and adjusted to 1x10 ⁵ cells/mL, and then 1 mL of prepared cells were added to a 24-well culture plate and cultured for 3-4 days. When the cells have grown to about 80%, they are treated with samples of various concentrations (50, 100, 500, 1000, and 2000 ㎍/mL) in medium without fetal bovine serum and antibiotics, and then cultured for an additional 24 hours. The survival rate of these cells was measured using the MTT reduction method.

Approximately one-tenth of the volume of the growth medium was added with MTT solution (5 mg/mL) to each well, and then cultured at 37°C for another 4 hours. Then, remove the medium, being careful to prevent the formazan produced by reducing MTT from leaving the medium. After leaving the sample at room temperature for 30 minutes to completely remove the remaining medium, dissolve the sample using DMSO and measure the absorbance at 570 nm. was measured. Cell survival rate was calculated according to Equation 1 below and is shown in Table 1 below.

[Equation 1]

Cell viability (%) = (absorbance of sample treatment group / absorbance of control group) × 100

division Cell viability (%) Sample processing concentration (㎍/mL) 50 100 500 1000 2000 Control group (untreated group) 100 Example 1 113.1 132.3 116.5 99.7 94.9 Example 2 121.2 141.4 123.1 110.3 96.6 Example 3 108.6 124.5 110.2 101.2 95.4 Example 4 108.7 126.1 109.9 102.4 93.8

As shown in Table 1, it was confirmed that the mixed fermentation extract according to the example of the present invention can be safely used on human skin up to 2,000 μg/mL.

Test Example 2: Antioxidant activity (DPPH radical scavenging ability)

After dissolving the test substance in distilled water, test solutions diluted to various concentrations were prepared.

100 μL of the test solution diluted to various concentrations and 100 μL of 0.15 mM DPPH (2,2-diphenyl-1-picrylhydrazyl) solution were added to a 96-well plate, mixed, and reacted at room temperature for 30 minutes. Then, the absorbance was measured at a wavelength of 570 nm. . The results showed the removal activity of free radicals compared to the control group to which no sample was added, and ascorbic acid was used as the positive control group. DPPH radical scavenging ability was calculated using the formula below, and the SC ₅₀ value was calculated from the concentration at which the radical scavenging ability is 50% from the calibration curve for the radical scavenging ability at each concentration and is shown in Table 2 below.

[Equation 2]

DPPH radical scavenging ability (%) = (1-A/B) x 100

(A: Absorbance of sample addition section, B: Absorbance of untreated section)

division DPPH radical scavenging ability (%) SC ₅₀
(㎍/mL) Sample processing concentration (㎍/mL) 50 100 200 300 500 control group
(Untreated group) 100 Example 1 27.7±2.5 39.5±4.8 55.1±3.3 64.4±5.9 69.8±6.3 166.5±9.4 Example 2 20.4±0.6 31.1±1.2 48.2±2.3 56.7±1.4 60.7±1.7 225.6±11.7 Example 3 33.1±1.2 44.7±3.5 60.4±4.7 69.7±5.2 76.3±0.7 137.8±16.3 Example 4 43.1±2.9 54.1±2.3 69.6±1.2 76.2±1.6 83.6±2.6 82.7±9.1 Comparative Example 1 13.7±1.1 19.8±0.7 26.2±1.5 32.1±2.4 35.7±2.0 - Comparative Example 5 16.7±1.3 25.4±1.9 40.6±2.8 51.2±3.7 54.8±1.5 290.4±21.6 Comparative Example 6 15.3±1.4 24.5±3.3 36.3±2.6 47.1±2.1 51.4±3.3 451.8±10.2

As shown in Table 2, it can be seen that the mixed fermentation extract according to the example shows a significantly superior antioxidant effect compared to the comparative example.

Test Example 3: Whitening effect

3-1: Tyrosinase activity inhibition ability

The ability to inhibit tyrosine hydroxylase activity of tyrosinase, which hydroxylates tyrosine to 3,4-dihydroxyphenylalanine (DOPA), was evaluated. The test substances were dissolved in distilled water and diluted in 0.1 M phosphate buffer (pH 6.5) to prepare test solutions of various concentrations. In a 96-well plate, sequentially add 170 μL of 0.1 M phosphate buffer (pH 6.5), 10 μL of test solution diluted to various concentrations, 10 μL of 2000 unit/mL mushroom tyrosinase, and 10 μL of 1.5 mM L-tyrosine, then mix and cool at 37°C. After reacting for 15 minutes, absorbance was measured at a wavelength of 490 nm. The absorbance of the blank test solution was measured by reacting in the same manner by adding 0.1 M phosphate buffer (pH 6.5) instead of the test solution. The inhibition rate of tyrosinase enzyme activity was calculated using the formula below and is shown in Table 3 below.

[Equation 3]

Enzyme activity inhibition rate (%) = (A-B)/A x 100

(A: Absorbance after reaction of blank test solution, B: Absorbance after reaction of test solution)

division Tyrosinase activity inhibition rate (%) Sample processing concentration (㎍/mL) 50 100 200 300 Control group (untreated group) 100 Example 1 19.3±3.0 39.1±0.5 58.5±1.3 62.5±1.3 Example 2 14.4±1.5 28.2±1.3 45.2±1.0 49.7±0.5 Example 3 23.2±2.2 48.6±1.2 67.6±0.6 71.7±1.0 Example 4 26.7±0.2 56.8±0.4 74.5±0.5 79.1±2.4 Comparative Example 1 11.5±2.3 21.2±1.5 29.3±2.7 34.6±0.5 Comparative Example 2 8.6±1.2 15.4±0.8 17.5±1.9 21.5±0.1 Comparative Example 3 7.5±2.0 11.3±0.7 15.1±0.5 17.5±0.6 Comparative Example 4 - 8.5±1.3 12.5±0.8 15.4±0.3 Comparative Example 5 10.3±1.3 19.5±2.2 27.7±2.6 31.6±1.9 Comparative Example 6 13.7±1.1 21.6±1.9 32.4±2.4 39.5±2.0

As shown in Table 3, the tyrosinase inhibitory effect of the mixed fermentation extract according to the example was confirmed to be very excellent.

3-2: Melamine synthesis inhibition ability

B16F10 cell culture

B16F10 cells, melanoma cells derived from mice, were grown in Dulbecco's modified Eagle's medium (DMEM, Gibco-Invitrogen, Carlabad, CA, USA) with 10% fetal bovine serum (FBS), 100 units/mL penicillin, and 100 μg/mL streptomycin. The added cell culture medium was used to culture in a humid CO ₂ incubator (5% CO ₂ /95% air) at 37°C. When the culture dish was about 80% filled with cells, the cell monolayer was washed with phosphate buffer saline (PBS, pH 7.4), cell culture medium was added, cells were detached and subcultured, and the medium was changed every two days.

Measurement of melanin content produced in B16F10 cells

B16F10 cells were distributed in a 12-well plate at 5 × 10 ⁴ cells/well and cultured for 24 hours. After culturing the cells for 24 hours, the cell culture medium was exchanged with a culture medium containing 100 nM α-melanocyte-stimulating hormone (α-MSH, Sigma-Aldrich) and various concentrations of test substances. After culturing the cells for 72 hours, the cell monolayer was washed with PBS, treated with 0.25% trypsin-2.65 mM EDTA to recover the cells, and centrifuged at 12,000 rpm for 10 minutes to obtain a pellet. 1 N NaOH containing 10% dimethylsulfoxide (DMSO) was added to the obtained pellet to dissolve it at 60°C, and the absorbance was measured at a wavelength of 490 nm. The amount of melanin was calculated from a calibration curve using synthetic melanin, the protein was measured using a BCA protein assay kit (Thermo Scientific), the amount of melanin produced was normalized to the amount of protein, and the value of the group treated only with α-MSH (stimulant) was calculated. It is expressed as a relative value by setting it to 100. At this time, the positive control group is the test group treated with 40 μM of Melasolv. For reference, the melanin content of the untreated group in which neither α-MSH nor the sample was treated was measured to be 44.1 ± 8.8%.

division Melanin content (% of control) Sample processing concentration (㎍/mL) 50 100 200 300 α-MSH only 100 Melasolve + α-MSH 39.6±4.7 Example 1 + α-MSH 89.9±2.4 78.7±6.2 64.2±1.5 55.0±2.7 Example 2 + α-MSH 95.3±1.5 86.6±2.1 73.1±0.7 64.2±1.6 Example 3 + α-MSH 86.2±1.3 74.5±2.7 58.5±2.4 47.3±2.1 Example 4 + α-MSH 83.1±1.2 68.1±0.8 52.6±0.3 41.1±1.6 Comparative Example 1 + α-MSH 98.4±0.6 95.2±2.7 89.1±1.2 83.3±1.5 Comparative Example 2 + α-MSH 99.5±1.3 98.3±1.1 95.6±2.9 91.6±2.2 Comparative Example 3 + α-MSH 98.2±2.6 97.3±2.0 95.1±2.6 90.3±3.1 Comparative Example 4 + α-MSH 98.9±2.1 96.1±1.3 92.2±1.5 87.4±1.3 Comparative Example 5 + α-MSH 98.1±0.2 96.2±1.8 92.3±0.7 85.1±0.3 Comparative Example 6 + α-MSH 92.9±1.4 89.2±2.0 83.4±3.8 76.7±2.6

As shown in Table 4, the melanin synthesis inhibition effect of the mixed fermentation extract according to the example was confirmed to be very excellent.

Test Example 4: Skin wrinkle improvement effect

4-1: Promotes synthesis of type 1 procollagen protein

HS68 cell culture

HS68 cells, human-derived skin fibroblasts, were purchased from the Korea Cell Line Bank. HS68 cells were grown in a cell culture medium (complete DMEM medium) supplemented with 10% fetal bovine serum (FBS), 100 units/mL penicillin, and 100 μg/mL streptomycin in DMEM medium (Welgene) at 37°C in a humidified CO ₂ incubator ( Cultured in 5% CO ₂ /95% air). When the culture dish was about 80% filled with cells, the cell monolayer was washed with phosphate buffer saline (PBS, pH 7.4), trypsin-2.65mM EDTA was added, cells were detached and subcultured, and the medium was changed every 2 days. .

Measurement of type 1 procollagen content produced and secreted by HS68 cells

HS68 cells were distributed in a 24-well plate at 1 × 10 ⁵ cells/well and the cells were cultured for 24 hours. After culturing the cells for 24 hours, they were replaced with serum-free cell culture medium containing various concentrations of each test substance, and the cells were cultured for 24 hours. After culturing the cells for 24 hours, the cell culture medium was collected and the procollagen type I C-peptide (PICP) content was measured according to the method suggested by the manufacturer using the Procollagen Type I C-Peptide (PIP) EIA Kit (Takara). was measured, and the relative values to the values of the untreated group in which the sample was not treated are shown in Table 5 below. At this time, the positive control group is the test group treated with 5 ng/mL of TGF-β1.

division Type 1 procollagen content (% of control) Sample processing concentration (㎍/mL) 50 100 200 300 500 Untreated group 100 TGF-β1 371.0±6.0 Example 1 246.6±22.7 255.1±20.7 271.8±15.9 275.4±0.7 282.9±5.1 Example 2 150.2±11.0 163.1±12.6 172.4±9.7 189.8±8.9 205.1±6.5 Example 3 267.7±13.8 284.3±3.4 309.4±6.7 316.8±9.1 325.5±1.9 Example 4 292.8±11.2 326.6±11.6 345.8±11.2 357.9±1.6 368.5±6.2 Comparative Example 1 103.6±2.3 109.2±10.4 116.4±11.5 115.9±10.1 125.6±4.7 Comparative Example 2 100.8±11.2 102.5±11.0 103.7±13.6 106.1±12.7 105.5±6.3 Comparative Example 3 131.2±10.5 148.6±12.4 155.4±9.5 172.2±10.2 186.6±8.1 Comparative Example 4 109.4±5.1 116.1±10.1 124.8±11.3 134.8±9.9 137.2±10.8 Comparative Example 5 134.8±13.5 156.4±11.0 162.8±16.1 186.1±7.6 195.1±7.3 Comparative Example 6 128.6±10.7 148.8±7.5 156.3±10.4 174.7±5.7 183.6±9.7

As shown in Table 5, it can be confirmed that the mixed fermentation extract according to the example effectively prevents the formation of wrinkles and improves the wrinkles formed through the action of promoting the production of type 1 procollagen.

4-2: Inhibition of synthesis of MMP-1 protein

HS68 cells were distributed in a 24-well plate at 1 × 10 ⁵ cells/well and the cells were cultured for 24 hours. After culturing the cells for 24 hours, they were replaced with serum-free cell culture medium containing various concentrations of each test substance, and the cells were cultured for 24 hours. After culturing the cells for 24 hours, the cell culture medium was collected and the MMP-1 content was measured using the Human MMP-1 ELISA Kit (Sigma-Aldrich) according to the method suggested by the manufacturer. The relative values are shown in Table 6 below. For reference, the MMP-1 content of the untreated group was measured to be 4.21 ± 0.10 ng/mL.

division MMP-1 content (% of control) Sample processing concentration (㎍/mL) 50 100 200 300 500 Untreated group 100 Example 1 64.6±2.7 58.1±2.7 54.8±1.9 51.4±0.7 48.3±5.1 Example 2 73.2±1.0 66.1±2.6 63.4±4.7 60.8±4.9 57.1±6.5 Example 3 58.7±3.8 49.3±3.4 46.4±3.7 45.8±4.1 42.5±1.9 Example 4 56.8±1.2 45.6±1.6 42.8±1.2 40.9±1.6 38.5±2.2 Comparative Example 1 93.6±2.3 89.2±10.4 83.4±11.5 82.9±10.1 83.6±4.7 Comparative Example 2 100.8±11.2 102.5±11.0 98.7±3.6 96.1±7.7 95.5±6.3 Comparative Example 3 88.2±10.5 83.6±1.4 75.4±9.5 72.2±10.2 70.6±5.1 Comparative Example 4 92.4±5.1 86.1±10.1 83.8±6.3 79.8±9.9 77.2±1.8 Comparative Example 5 84.8±3.5 78.4±8.0 72.8±6.1 70.1±7.6 68.1±1.3 Comparative Example 6 86.5±2.6 80.1±3.4 75.9±2.7 72.5±6.1 68.6±2.4

As shown in Table 6, it can be confirmed that the mixed fermentation extract according to the example effectively prevents the formation of wrinkles and improves the wrinkles formed through the action of suppressing MMP-1 expression.

<Production Example 1> Production of cosmetics (essence)

Essences were prepared with cosmetic compositions to which the mixed fermentation extracts according to Examples and Comparative Examples were applied, respectively, with the compositions shown in Tables 7 and 8 below.

Ingredients Content (% by weight) Manufacturing Example 1-1 Manufacturing Example 1-2 Manufacturing Example 1-3 Manufacturing example
1-4 Example 1 0.2 - - - Example 2 - 0.2 - - Example 3 - - 0.2 - Example 4 - - - 0.2 Triethanolamine 0.25 0.25 0.25 0.25 Carboxyvinylpolymer ⁽¹⁾ 0.22 0.22 0.22 0.22 glycerin 4.0 4.0 4.0 4.0 propylene glycol 0.2 0.2 0.2 0.2 Paraoxybenzoic acid
ester 0.2 0.2 0.2 0.2 beeswax 0.5 0.5 0.5 0.5 Cetostearyl alcohol 1.0 1.0 1.0 1.0 Glyceryl Mono
stearate 1.0 1.0 1.0 1.0 monostearic acid
Sorbitan ⁽²⁾ 0.5 0.5 0.5 0.5 monostearic acid
polyethylene glycol 8.0 8.0 8.0 8.0 Propyl paraoxybenzoic acid 0.1 0.1 0.1 0.1 dimethylsiloxane 0.3 0.3 0.3 0.3 isocetyl
Octanoate ⁽³⁾ 3.0 3.0 3.0 3.0 squalane 5.0 5.0 5.0 5.0 Spices Appropriate amount Appropriate amount Appropriate amount Appropriate amount Distilled water remaining amount remaining amount remaining amount remaining amount

* Note ¹ : Carboxyvinyl polymer - 1% by weight aqueous solution of Carbopol 941 from BFGoodrich, USA * Note ² : Sorbitan monostearate - Arlacel 60 from ICI, UK 60)

* Note ³ : Isocetyl octanoate - ICEH from Nihon KoKyu Alcohol, Japan

Ingredients Content (% by weight) Comparative Manufacturing Example 1 Comparative Manufacturing Example 2 Comparative Manufacturing Example 3 Comparative Manufacturing Example 4 Comparative Manufacturing Example 5 Comparative Manufacturing Example 6 Comparative Example 1 0.2 - - - - - Comparative Example 2 - 0.2 - - - - Comparative Example 3 - - 0.2 - - - Comparative Example 4 - - - 0.2 - - Comparative Example 5 - - - - 0.2 - Comparative Manufacturing Example 6 - - - - - 0.2 Triethanolamine 0.25 0.25 0.25 0.25 0.25 0.25 Carboxyvinyl polymer (1) 0.22 0.22 0.22 0.22 0.22 0.22 glycerin 4.0 4.0 4.0 4.0 4.0 4.0 propylene glycol 0.2 0.2 0.2 0.2 0.2 0.2 Paraoxybenzoic acid
ester 0.2 0.2 0.2 0.2 0.2 0.2 beeswax 0.5 0.5 0.5 0.5 0.5 0.5 Cetostearyl alcohol 1.0 1.0 1.0 1.0 1.0 1.0 Glyceryl Mono
stearate 1.0 1.0 1.0 1.0 1.0 1.0 monostearic acid
Sorbitan (2) 0.5 0.5 0.5 0.5 0.5 0.5 monostearic acid
polyethylene glycol 8.0 8.0 8.0 8.0 8.0 8.0 Propyl paraoxybenzoic acid 0.1 0.1 0.1 0.1 0.1 0.1 dimethylsiloxane 0.3 0.3 0.3 0.3 0.3 0.3 isocetyl
Octanoate (3) 3.0 3.0 3.0 3.0 3.0 3.0 squalane 5.0 5.0 5.0 5.0 5.0 5.0 Spices Appropriate amount Appropriate amount Appropriate amount Appropriate amount Appropriate amount Appropriate amount Distilled water remaining amount remaining amount remaining amount remaining amount remaining amount remaining amount

Test Example 5: Skin moisturizing effect

The clinical moisturizing effect and durability of the cosmetic compositions of Preparation Examples and Comparative Preparation Examples were measured as follows.

Through a survey, 120 healthy adult men and women who felt that their skin was dry were selected and randomly divided into 12 groups (10 people each), and each group was given Preparation Examples 1-1 to 1-6 and Comparative Preparation Examples 1 to 1. The essence prepared according to 5 was applied to the entire face twice daily (morning/evening) for 30 days. At this time, before applying the essence, measure the amount of skin moisture using a moisture meter (Corneometer, CM825 courage Khazaka electronic GmbHtk, Germany) under constant temperature and humidity conditions (24 ℃, humidity 40%), and set this as the default value, 30 After one day had elapsed, the skin moisture content was measured and the amount of change was evaluated. For reference, the measurement value of the moisture meter increases as the skin moisture content increases, and the measurement coefficient is arbitrary unit (A.U.). The experimental results are shown in Table 9 below.

division Before application After 30 days Skin moisture increase rate (%) Control group (untreated group) 25.76 26.03 1.05 Manufacturing Example 1-1 24.95 45.62 84.26 Manufacturing Example 1-2 24.20 39.52 63.31 Manufacturing Example 1-3 25.17 49.19 95.45 Manufacturing Example 1-4 25.06 51.80 107.21 Comparative Manufacturing Example 1 23.90 38.18 59.75 Comparative Manufacturing Example 2 23.50 31.09 32.30 Comparative Manufacturing Example 3 23.22 25.64 10.42 Comparative Manufacturing Example 4 25.23 34.46 36.58 Comparative Manufacturing Example 5 23.19 34.41 48.37 Comparative Manufacturing Example 6 24.25 36.42 50.19

As shown in Table 9, it was confirmed that the cosmetic composition according to the Preparation Example had significantly higher skin moisturizing effect and durability compared to the cosmetic composition of the Comparative Preparation Example.

Test Example 6: Human patch test

The human patch test was conducted in accordance with the ethical regulations based on the Declaration of Helsinki and the Ministry of Food and Drug Safety Cosmetic Human Application Testing Guidelines, using the method devised by Frosch & Kligman (J. Am. Acad. Dermatol, 1(1) 35 (1979) ) was applied to 48 healthy female subjects aged 20 to 60 (average age 41.34 ± 8.06 years) without specific skin diseases or allergies. First, the patch area was disinfected with 70% ethanol, and then a Van der Bend (Van der Bend, Netherlands) patch containing 20 μl of the samples (5.0 mg/mL) of Examples 1 to 6 was applied. After 48 h, the patch was removed, the test site was marked with a skin marker (Chemotechnique Diagnostics AB, Sweden), and the reactivity of each test site was evaluated after 20 min and 24 hours. The evaluation criteria for the primary skin irritation test were in accordance with the PCPC Guidelines shown in Table 10 below, and the degree of skin reaction (average reactivity) evaluated twice was calculated according to Equation 4 below: Scored.

Responsiveness Clinical symptoms (reaction) 0 No response One faint or mild erythema 2 Moderate erythema; and barely detectable swelling and/or papule formation at the patch border. 3 Moderate erythema; and edema spread throughout the patch area. 4 severe erythema; and severe edema; or blisters 5 Severe reactions spread beyond the patch site

[Equation 4]

Average reactivity = [(Reactivity × Number of subjects who responded)/(Total number of subjects × Highest score (5 points)) × 100] × 1/2

In order to evaluate the degree of skin irritation on human skin, a human patch test was performed by applying 20 μL of the samples of Examples 1 to 6 in liquid state to each of 8 subjects (48 in total), and the results are shown in Table 11 below. indicated.

division Number of subjects who responded (people) Responsiveness 24 hours 48 hours average negative control 0 0.0 0.0 0.0 Example 1 0 0.0 0.0 0.0 Example 2 0 0.0 0.0 0.0 Example 3 0 0.0 0.0 0.0 Example 4 0 0.0 0.0 0.0

Looking at Table 11, as a result of evaluating the degree of skin irritation by applying the composition according to the embodiment of the present invention, all 48 subjects showed a negative reaction with a reaction grade of 0.0, so the composition of the present invention was a primary irritant to human skin. From this perspective, it is considered a non-irritating material.

The following illustrates a preparation example for the mixed fermented extract of wisteria flowers of the present invention.

<Production Example 2> Production of cosmetics

<2-1> Soft lotion (skin)

In order to produce a softening lotion containing the mixed fermented extract of wisteria flowers of the present invention for antioxidant purposes, for skin moisturizing, for improving skin acne, or for improving skin troubles, it is mixed as shown in Table 12 below and follows a conventional manufacturing method in the field of cosmetics. It can be manufactured according to.

ingredient Content (% by weight) Wisteria flower mixed fermentation extract of Example 1 0.2 glycerin 6.0 1,3-butylene glycol 3.0 PEG 1500 1.0 allantoin 0.1 DL-panthenol 0.3 EDTA-2Na 0.02 Benzophenone-9 0.04 Sodium Hyaluronate 5.0 ethanol 10.0 Polysorbate 20 0.2 Preservatives, fragrance, coloring a very small amount Distilled water remaining amount Sum 100

<2-2> Nourishing lotion (lotion)

To produce a nutritional lotion containing the mixed fermented extract of wisteria flowers of the present invention for antioxidant purposes, for skin moisturizing, for improving skin acne, or for improving skin troubles, it was mixed as shown in Table 13 below and used in a conventional manufacturing method in the cosmetics field. It can be manufactured according to.

ingredient Content (% by weight) Wisteria flower mixed fermentation extract of Example 1 0.2 propylene glycol 6.0 glycerin 4.0 Triethanolamine 1.2 Tocopheryl Acetate 3.0 liquid paraffin 5.0 squalane 3.0 macadami nut oil 2.0 Polysorbate 60 1.5 Sorbitan sesquioleate 1.0 Carboxy vinyl polymer 1.0 Preservatives, fragrance, coloring a very small amount Distilled water remaining amount Sum 100

<2-3> Nutritional cream

In order to produce a nutritional cream for antioxidant, skin moisturizing, improving skin acne or improving skin troubles containing the mixed fermented extract of wisteria flowers of the present invention, it was mixed as shown in Table 14 below and used in a conventional manufacturing method in the cosmetics field. It can be manufactured according to.

ingredient Content (% by weight) Wisteria flower mixed fermentation extract of Example 1 0.2 Lipophilic glycerin monostearate 1.5 cetearyl alcohol 1.5 stearic acid 1.0 Polysorbate 60 1.5 Sorbitan Stearate 0.6 Isostearyl isostearate 5.0 squalane 5.0 mineral oil 35.0 Dimethicone 0.5 Hydroxyethylcellulose 0.12 glycerin 6.0 Triethanolamine 0.7 Preservatives, fragrance, coloring a very small amount Distilled water remaining amount Sum 100

<2-4> Emulsion for mask pack

In order to prepare a mask pack emulsion containing the mixed fermented extract of wisteria flowers of the present invention for antioxidant purposes, for skin moisturizing, for improving skin acne, or for improving skin troubles, the mixture is mixed as shown in Table 15 below, and a manufacturing method in the conventional cosmetics field is used. It can be manufactured according to.

ingredient Content (% by weight) Wisteria flower mixed fermentation extract of Example 1 0.2 polyvinyl alcohol 15.0 cellulose gum 0.15 glycerin 3.0 PEG 1500 2.0 Cyclodextrin 0.15 DL-panthenol 0.4 allantoin 0.1 Monoammonium glycyrrhizinate 0.3 nicotinamide 0.5 ethanol 6.0 PEG 40 Hydrogenated Castor Oil 0.3 Preservatives, fragrance, coloring a very small amount Distilled water remaining amount Sum 100

<Manufacturing Example 3> Manufacturing of health functional food

<3-1> Manufacturing of health drinks

A health drink containing the mixed fermented wisteria flower extract of the present invention with the ingredients and contents shown in Table 16 below was prepared according to a conventional method.

Mix the ingredients in Table 16 according to a typical health drink manufacturing method, stir and heat at 85°C for about 1 hour, filter the resulting solution, place it in a sterilized 2 L container, seal, sterilize, and store in the refrigerator. Used in the production of the functional beverage composition of the present invention.

The composition ratio is a preferred embodiment of mixing ingredients that are relatively suitable for beverages of preference, but the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as demand class, country of demand, and intended use.

ingredient content Wisteria flower mixed fermentation extract of Example 1 500mg citric acid 1000mg oligosaccharide 10 g plum concentrate 2.0g taurine 1.0 g Purified water remaining amount Sum 100g

Although the present invention has been described in terms of the above-mentioned preferred embodiments, various modifications and variations can be made without departing from the gist and scope of the invention. Additionally, the appended claims cover such modifications or variations as fall within the subject matter of the present invention.

Claims (7)

Contains fermented extracts of wisteria flowers, aralia flowers, camellia leaves, and camellia flower mixtures as active ingredients,
The mixture includes wisteria flowers, aralia flowers, camellia leaves and camellia flowers in a weight ratio of 1:0.05-0.5:0.2-2:0.1-1,
A cosmetic composition for antioxidant, skin whitening, and skin wrinkle improvement, wherein the fermentation is performed using Lactobacillus rhamnosus. According to paragraph 1,
A cosmetic composition, characterized in that the fermented extract of the mixture is contained in an amount of 0.0001 to 20% by weight based on the total weight of the cosmetic composition. delete According to paragraph 1,
A cosmetic composition characterized in that the mixture further comprises 10 to 20 parts by weight of Ubangja based on 100 parts by weight of the mixture. delete The cosmetic composition according to claim 1, wherein the fermented extract of the mixture is obtained by fermenting the mixture and then extracting it under ultra-high pressure. According to paragraph 1,
A cosmetic composition characterized in that the fermented extract of the mixture exhibits the following activities (1) to (5):
(1) activity to suppress expression of MMP-1 protein;
(2) activity to promote the expression of procollagen type I protein;
(3) tyrosinase activity inhibitory activity;
(4) Melamine synthesis inhibitory activity; and
(5) DPPH (1-1-diphenyl-2-picryl-hydrazyl) radical scavenging activity.