KR102460358B1 - Cosmetic composition comprising fermented herbs mixture extract for preventing or improving of skin whitening, anti-wrinkle, UV-protective, anti-atopic and tinea pedis - Google Patents

Abstract

The present invention relates to a cosmetic or quasi-drug composition containing a herbal mixed extract by selectively using plant-derived lactic acid bacteria as a fermenting strain and containing it as an active ingredient. A cosmetic composition having whitening, wrinkle improvement, UV protection, anti-atopic and athlete's foot prevention or improvement functions containing the extract as an active ingredient by experimentally confirming the whitening, wrinkle improvement, UV protection, anti-atopic and athlete's foot prevention or improvement effects of the extract; It has an excellent effect of providing quasi-drugs.

Description

Whitening, wrinkle improvement, UV protection, anti-atopic and athlete's foot prevention or improvement functional cosmetic composition containing lactic acid bacteria fermented herb mixture extract for preventing or improving of skin whitening, anti-wrinkle, UV-protective, anti-atopic and tinea pedis}

The present invention relates to a functional cosmetic composition for whitening, wrinkle improvement, UV protection, anti-atopic and athlete's foot prevention or improvement using herbal plants, and more particularly, rosemary, lavender, sage, basil, mint, chamomile, oregano, among herbal plants. It relates to a whitening, anti-wrinkle, UV protection, anti-atopic and athlete's foot prevention or improvement functional cosmetic composition containing an herbal extract obtained by fermenting lemon balm and stevia with an herbal-derived lactic acid strain.

The word herb has its etymology from the Latin word 'herba', which means green grass, and was used in ancient countries to mean incense and medicinal herbs. It is defined as “a plant whose leaves or stems are used for food or medicinal purposes, or used for incense and flavor”. In other words, herbs are fragrant and useful plants for humans. Since ancient times, herbs have played an important role as medicinal herbs for antiseptic and insecticide as well as for pain relief and sedation. and antibacterial ([Naver Knowledge Encyclopedia] Herb [Herb] (Cooking Glossary, 2007. 8. 27., Gwangmungak)).

Rosemary is a perennial evergreen deciduous tree in the family Lamiaceae, which usually grows over 1 meter. The narrow, thin needle-shaped leaves have a leather-like quality, are shiny, have a characteristic strong aroma, and are dark green. It has sterilization and disinfection action, so it was used to preserve meat, and it was often used in hospitals or when infectious diseases were prevalent. Rosemary helps to improve memory, revitalizes the symptoms of lethargy, and relieves headaches and migraines. In addition, it has an anti-obesity effect, regenerates the skin, revitalizes the sagging tissue, suppresses the occurrence of edema and dandruff, and promotes hair growth. Rosemary is used for cooking, facial cleansing, and to increase blood pressure when you have low blood pressure.

Lavender is one of the most well-known herbs. High-quality lavender is used for many purposes, especially flowers, leaves, stems, and the like, so it is suitable for ornamental purposes as well as potpourri and various cosmetic products. The lotion made with the essential oil of lavender is effective for rough skin because it has a cleansing effect that relaxes and relieves the skin and regenerates it clean and moist. Also, the scent of lavender has a calming effect, so it was put in a pillow and used for sleep. Brewed tea with lavender is effective for sedation, relieves pain and headaches, and helps you sleep better by changing your mood.

Sage has been used as a panacea since the Greek and Roman times. It is used in various ways such as ornamental, spice collection, medicine, cooking, dyeing, etc., and has tonic, soothing, digestive and sterilizing effects. Sage has many uses, but should not be used in large quantities at once. Sage tea is effective in clearing the mood, calming the excitement, and preventing stomatitis, bleeding gums, and bad breath. In addition, sage promotes digestion, detoxifies, placebo, and baths are used for fatigue recovery, expectoration, gout, diuresis, anthelminth, and antiseptic, and has a great effect on poor circulation and menopausal disorders.

As an essential flavoring agent in Basil cuisine, it is used in dishes such as chicken, seafood, vegetables, salad, spaghetti, pizza pie, stew, soup, and sauce. Dried basil has medicinal effects to clear the head and eliminate headaches, so drinking it as a tea has irritability and headache as well as stomatitis and tonic effects.

Mint is a native of Europe, and along with lemon balm, it can be said to be a representative herb of the milwon plant. It is a perennial plant of the Lamiaceae family and one of the strongest herbs to grow. It has a strong mint scent that gives a refreshing and refreshing feeling just by rubbing the leaves. When storing, cut off the stems of the stems in June or July, when the scent is strongest, and dry them in the shade. Mint herbs, characterized by their refreshing feeling, are especially used to make sauces for meat dishes, and with their refreshing taste and scent, they are used to flavor beanies, carbonated water drinks, cocktails, etc. Although there are some differences depending on the type of mint, it is used as a raw material for sterilization, digestion promotion, gastric action, oral deodorant, toothpaste, and placebo. It has medicinal effects on dermatitis and itching, and when placed in the bathroom, the mint scent lasts for a long time instead of the bad smell, and it even has a strong sterilization effect.

Chamomile is one of the most well-known herbs as a medicinal herb among many herbs. It grows widely in Europe, India and North Africa and belongs to the Asteraceae family. It smells like sweet and sour apples, but in spring, tiny flowers bloom all at once, and the sweet fragrance spreads throughout. Dried flowers have excellent cosmetic effects and are used as a bathing agent. If you add a small amount to massage oil, it is good for recovery from fatigue after exercise. It is good for low inflammation, antiseptic, anthelmintic, and convulsions. In addition, it has the effect of warming the body for those who have a cold.

Oregano (Oregano) is a perennial plant of the leaf family, and its origin is Europe. Also called flower mint, it has a pungent mint-like scent. It is harvested during the flowering period, dried and preserved, and dried leaves are used as spices. Oregano extract, which has been used as a medicinal herb since the ancient Greek times, has tonic, diuretic, gastrointestinal, appetite-enhancing, calming, and sterilizing effects.

Lemon Balm is also called 'Melissa' and has been famous as a wheat source plant since ancient times. It has a lemon-like scent, which calms emotions, lowers heart rate and lowers blood pressure. When bitten by a scorpion or tarantula, it has a detoxifying effect, relieves diarrhea, and is very effective in preventing viruses. It is also effective against smallpox and has been used for mouth sores when there is a toothache. It has been used for detoxification of poisonous mushrooms, stomach pain, stomach ulcer, suppression of menstrual pain, and promotion of menstruation.

Stevia grows in the mountainous regions of South America, such as Paraguay, Argentina, and Brazil. In Paraguay, stevia leaves have been used as a sweetener since ancient times. The leaves contain stevioside, a sweetener, about 6-7% by weight, and the content varies greatly depending on the individual. The sweetening ingredient is 300 times that of sugar, and it is used as a sweetener in soft drinks, such as making tea or as a substitute for chewing gum. The leaves are steamed and dried and used as a sweetener in diet foods such as sherbet, ice cream, and chewing gum.

Lactic acid bacteria are safe microorganisms that are easily found in the intestines of humans and animals and in fermented foods (Orrhge, K. et al., 2000, Bifidobacteria and lactobacilli in human health, Drugs Exptl. Clin. Res ., 26, 95-111) It attaches to intestinal epithelial cells and becomes parasitic to improve the properties of the intestinal flora, thereby stabilizing the intestinal flora, inhibiting the settling of harmful bacteria and reducing the production of decay products and preventing diseases, activating the immune system, and having an anticancer effect It is known to help the body a lot, such as lowering cholesterol. The reason that lactic acid bacteria has a growth inhibitory action against various putrefactive microorganisms and pathogenic microorganisms is due to several metabolic characteristics. Ida (Fuller, K., 1989, Probiotics in man and animals, J. Appl. Bacteriol., 66, 365-378).

In general, ultraviolet rays included in the sun's rays are divided into long wavelength, medium wavelength, and short wavelength according to the wavelength region, and these ultraviolet rays are known to cause various side effects on the skin. Short-wavelength ultraviolet rays among ultraviolet rays are very harmful to the human body, but so far they are absorbed by ozone insects in the atmosphere and hardly reach the earth's surface. , photoallergy, etc., and medium-wavelength ultraviolet rays with higher energy cause erythema, blisters, burns, skin cancer, pigmentation, and skin aging. very important. On the other hand, substances with ultraviolet absorbing action extracted and separated from natural products are generally less irritating and have other pharmacological actions such as anti-inflammatory, astringent, and antibacterial actions, so they have been promising as UV absorbers formulated in cosmetics. Efforts have been made to search for and separate UV-absorbing substances contained in natural products.

Atopic dermatitis is a chronic, recurrent inflammatory skin disease that mainly begins in infancy or childhood, and is accompanied by pruritus (itch), dry skin, and characteristic eczema. In infancy, it begins as eczema on the face and on the outstretched side of the limbs, but as you grow older, it characteristically appears in the form of eczema on the bent part of the arm and the bent part of the knee, and in many cases, it tends to improve naturally with growth. . In the case of adults, lichenification, which thickens the skin at the fold, appears, and eczema occurs on the face more often than in childhood. Atopic dermatitis is increasing worldwide, and there is a report that the prevalence is 20% of the population. The cause of atopic dermatitis is still unknown. Because clinical symptoms also appear in various ways such as dry skin and eczema, the cause of the onset cannot be explained by just one, but environmental factors, genetic predisposition, immunological reactions, and abnormal skin barrier are considered to be the main causes. Environmental factors include environmental pollution such as soot due to industrialization, increased use of food additives, increased use of carpets, beds, and sofas due to Western-style housing, and substances that cause allergies such as house dust mites due to increased indoor temperature (allergens) ) increase ([Naver Knowledge Encyclopedia] [atopic dermatitis] (Seoul National University Hospital Medical Information, Seoul National University Hospital)).

In order to improve atopic skin, conventionally, by cleaning the skin, removing substances that can act as antigens, strengthening the skin barrier function, and supplying ceramide, an intercellular lipid component necessary to protect the skin from drying, is used to prevent irritation to the stratum corneum. method has been used So, in order to improve atopic skin, well-known components such as ceramide, linoleic acid, vegetable oil or mineral oil, steroid preparations such as hydrocortisone, or specific plant extracts are included in the formulation of general emulsified products, or lecithin, a phospholipid component It is made in the form of liposomes and supplied to the stratum corneum of the skin to create an oil film to prevent moisture loss and moisture loss, and to prevent bacterial contamination. Although these methods have achieved some success in the prevention and treatment of atopic skin, they have several problems in terms of effectiveness and skin stability. For example, it has been reported that the use of steroids such as hydrocortisone may cause side effects such as epidermal growth inhibition or side effects, and urea peroxide, etc., irritates the skin and may cause another contact dermatitis. there is a problem. Therefore, in recent years, various studies on cosmetics for improving the condition of atopic skin or external preparations for skin are being actively conducted, and in particular, interest in natural substances that can be obtained from nature is increasing.

Athlete's foot (tinea pedis) is one of the most commonly observed symptoms of a fungal infection, and is a skin disease in which a fungus dissolves dead skin cells and uses them as nutrients to parasitize and reproduce. Among the fungi having affinity for the skin, their causative bacteria are mainly Trichophyton rubrum, Trichophyton mentagrophytes, Microsporum canis, Candida albicans, etc. It is said that they penetrate into the keratin of the skin and cause symptoms such as itching, redness, and inflammation. It is known that the degree of exacerbation of these fungal infections depends on the environment at the site of infection and individual immunity. On the other hand, in modern society, people mainly wear shoes or socks for a long time. After infection with the fungus as described above, if the skin is kept closed for a long time by shoes, socks, etc., the growth of fungi due to heat and moisture in the infected area This is improved, and athlete's foot occurs.

Athlete's foot develops in most parts of the body in places that are rich in dead skin cells that mold likes, moist, and warm (toes, soles of the feet, toenails, nails, sides, groin area, and places where the skin overlaps). In general, it is easy to think that only those that occur on the feet are athlete's foot, but it includes not only the feet and hands but also the genital warts around the groin; do. Statistically, more than 10% of the total population suffers from athlete's foot, and it occurs most often in people in their 20s and 40s, and rarely even in children.

According to the type of occurrence, athlete's foot is divided into interphalangeal type, vesicular type and keratinization type. The interphalangeal type mainly occurs between the toes, because this area is not well-ventilated with the outside air and has high moisture, so it is easy for athlete's foot bacteria to inhabit. When interdigital athlete's foot occurs, itching is severe, and the skin is whitened and cracked, giving off a lot of unpleasant odor, and sometimes it spreads to both toes and soles of the feet. It is known that blisters of various sizes are formed in the vesicular type athlete's foot, and when scratched due to itching, the skin is peeled off and scabs are frequently generated, and it is known that it worsens when there is a lot of sweat in summer. Keratinized type athlete's foot refers to a symptom in which the keratin becomes thick all over the sole of the foot and the powder falls off when it is thick, and it is easy to develop into chronic intractable athlete's foot.

In order to treat athlete's foot, a method of killing the causative agent of athlete's foot or inhibiting the growth of the bacteria is considered. The main treatment is antifungal, which is applied, and is widely used because it is convenient to use and has few side effects. However, it is difficult to cure only by using an applied therapeutic agent, and it is generally necessary to apply it for at least several weeks. Next, there is an antifungal drug that is eaten, and the effects of the above therapeutic agents are better than that of an applied drug, but side effects such as hepatotoxicity and nephrotoxicity do not occur infrequently, so care must be taken in administration. In the treatment of athlete's foot, keratin emollients are used together to exfoliate the dead skin cells in which the athlete's foot fungus lives, or steroid preparations are also used to improve symptoms. It is also commercialized.

The main ingredients of the athlete's foot drug on the market are largely divided into two categories: terbinafine and clotrimazole. Terbinafine has an excellent sterilization effect and has the advantage of being able to see the effect quickly in a short time. On the other hand, clotrimazole is focused on slow treatment by preventing the growth of bacteria rather than sterilization. Although the treatment period is long when clotrimazole is used, it is known to have strengths compared to products using terbinafine in restoring damaged skin. The above drugs are mainly prepared in a cream type or an ointment type, but they are good for use in symptoms that become hard like calluses as the keratin occurs severely. However, it is known that in the case of athlete's foot with oozing symptoms, secondary infection caused by skin disease or eczema along with athlete's foot, or toenail athlete's foot, the above drugs do not cure well. In addition, the ingredients of a wide range of skin disease treatments widely distributed in the market have been controversial because they contain corticosteroids that suppress normal immune responses in addition to ingredients that kill athlete's foot.

In relation to the present invention, the prior art on lactic acid bacteria is based on the cytotoxicity test and the antioxidant activity test of Lactobacillus rhamnosus by Choi et al. It has been confirmed that it has a high whitening cosmetic activity through a whitening efficacy experiment, and thus the possibility of using it as an antioxidant and whitening functional cosmetic material has been suggested (Cited documents: Choi, W.S, Kwon, H.S, Lim, H.W, No, R.W, Lee, H.Y. (2013) Korea J. Miceobiol. Biotechnol. 41(2), 183-189). Meanwhile, Korean Patent Registration No. 10-1178217 has been published for a composition for treating intestinal diseases and a composition for enhancing immunity, including Lactobacillus platarum CJLP243 KCCM11045P. In addition, a cosmetic with antioxidant, whitening and anti-wrinkle effects containing lactic acid bacteria fermented as an active ingredient, obtained by fermenting a mixed extract of pomegranate flower, pomegranate root, hollyhock flower, paprika, plum flower, plum tree root, lotus, and lotus root with lactic acid bacteria. A patent on the composition has been disclosed as Republic of Korea Patent No. 10-1452770, and has a skin protection effect against UV rays, whitening and anti-wrinkle effects, such as licorice, horseradish, hawthorn, black sesame, situation mushroom, angelica, sangbaekpi, white peony, and ginseng. , Korean Patent Registration No. 10-0966835, which relates to a cosmetic composition containing an extract of a plant mixture composed of gold, and a method for manufacturing the same has been published.

In addition, Kim et al. investigated the anti-atopic efficacy of yeast fermented products fermented with Lactobacullus bifermentans in 2,4-dinitrochlorbenzene-induced atopic dermatitis NC/Nga mice to develop substances that can improve symptoms of atopic dermatitis. The tendency to reduce atopic dermatitis by application of water was confirmed and it was reported that yulpi fermented product was effective in improving the symptoms of atopic dermatitis (cited documents: Kim, B.J, Son, W.R, Choi, M.O, Jo, S.K, Jung). , H.K, Lee, J.T, Kim, H.Y, Kweon, D.J. (2013) J Korea Soc Food Sci Nutr 42(9), 1378-1386), purslane extract, silk extract, arborvitae extract and cypress extract A functional cosmetic composition for alleviating and improving atopic symptoms containing the above plant extracts and a manufacturing method thereof have been disclosed in Korean Patent Registration No. 10-1516002.

As a prior art related to the prevention and improvement of athlete's foot, a composition for topical application for the prevention or treatment of athlete's foot comprising castor oil extracted with hexane, undecylenic acid, propyl alcohol, propylene glycol and water is disclosed in Korean Patent Registration No. 10-1281470 A composition for treating athlete's foot and eczema prepared by mixing and aging mugwort, yew, plantain, sagebrush, yellow lily, dermis, and hyangbuja in vinegar and wood vinegar is disclosed as Korean Patent Laid-Open Publication No. 10-2011-0014367 has been However, nowhere in the above literature, rosemary, lavender, sage, basil, mint, chamomile, oregano, lemon balm and stevia 9 kinds of herbal plants are selected and used to obtain extracts fermented with lactic acid bacteria, whitening, wrinkle improvement, UV protection, There is no disclosure with respect to a functional cosmetic composition for preventing or improving anti-atopic and athlete's foot.

Accordingly, an object of the present invention is to provide a functional cosmetic composition for whitening, wrinkle improvement, UV protection, anti-atopic and athlete's foot prevention or improvement containing a herbal mixed extract fermented using a plant lactic acid strain.

The above object of the present invention comprises the steps of preparing a raw material for fermentation by drying and powdering herbs and then sterilizing at a high temperature; Inoculating a vegetable lactic acid strain on the fermentation raw material obtained in the above step, and fermenting for 3 to 5 days under fermentation conditions of a temperature of 35 to 37 ℃, a pH of 7, and a stirring speed of 100 to 120 rpm to obtain an herbal fermented product; centrifuging the fermented product obtained in the above step and recovering only the supernatant; adding the same amount of an extraction solvent together with distilled water to the supernatant recovered in the above step, then extracting the distilled water with hot water to recover the distilled solution, and then obtaining the remaining lactic acid bacteria fermented extract; Filtering the lactic acid bacteria fermented extract remaining in the above step with a filter paper to prepare the fermented herbal extract of the present invention, confirming the whitening, wrinkle improvement, UV protection, anti-atopic and athlete's foot prevention or improvement effects of the extract, and applying the cosmetic composition It was achieved through the manufacturing steps.

Whitening, wrinkle improvement, UV protection, anti-atopic and athlete's foot prevention or improvement function containing the same as an active ingredient by confirming the whitening, wrinkle improvement, UV blocking, anti-atopic and athlete's foot prevention or improvement effects of the lactic acid bacteria fermented herbal extract according to the present invention There is an excellent effect of providing a cosmetic composition.

1 is a diagram showing a method for preparing a fermented herbal extract of a plant lactic acid strain of the present invention.

The method for producing a fermented herbal extract of the present invention is characterized in that it consists of the following steps (Fig. 1). Herbs suitable for achieving the object of the present invention are any one or more combinations selected from the group consisting of rosemary, lavender, sage, basil, mint, chamomile, oregano, lemon balm, and stevia.

Step 1: Prepare raw materials

Put the same amount of distilled water as the dried herb powder into a fermenter, sterilize it at 100℃ for 5 minutes, and then lower the temperature to 37℃ to prepare fermentation raw materials.

Step 2: Fermentation of plant lactic acid bacteria

The vegetable lactic acid strain cultured in liquid form in the lactic acid bacteria culture medium is inoculated into the fermentation raw material of the first step and fermented for 3 to 5 days at a temperature of 35 ~ 37 ℃, pH 7, and agitation speed of 100 ~ 120 rpm.

Step 3: Preparation of lactic acid bacteria fermented herbal extract

The fermented product obtained in the second step is centrifuged to remove sediment and cells, and only the fermentation supernatant is recovered. An extraction solvent is added to the fermentation supernatant together with the same amount of distilled water, and the distilled water is recovered by heating it at a temperature of 80° C. for 6 hours in an extractor equipped with a cooling condenser to obtain the remaining fermented extract. The lactic acid bacteria fermented herbal extract obtained above is first filtered with a 300 mesh filter paper, left at room temperature for 1 week, and then filtered with a 150 mm filter paper to prepare the lactic acid bacteria fermented herbal extract of the present invention.

As the herb of the first step, any one or more selected from the group of rosemary, lavender, sage, basil, mint, chamomile, oregano, lemon balm and stevia may be used, and most preferably, 4 or more kinds of herbs are used in the same amount (w/w ) is used in combination.

The plant lactic acid bacteria of the second step can be used without limitation as long as they are of the genus Lactobacillus, but Lactobacillus plantarum (L. plantarum, KFCC-11028), Lactobacillus sakei (L. sakei, KFCC 11383P), Lactobacillus lactis (L) It is preferable to select one of lactice, KCCM 11159P). The 'plant lactic acid bacteria' is a lactic acid strain characterized in that the raw material from which the lactic acid bacteria is separated is derived from the root thimble of a plant, that is, a herb.

In addition, as the lactic acid bacteria culture medium of the second step, MRS (Man Rogosa Sharpe), lactose, YM (Yeast and Mold), M17 or ART (Asparagine Enrichment Broth) medium may be used, but it is preferable to use MRS medium.

In addition, the inoculation amount of the lactic acid strain of the second step is 1x10 ⁸ ~ 5x10 ⁸ cells compared to 1 kg of the herbal plant of the first step.

Distilled water or an organic solvent may be used as the extraction solvent in the third step, but most preferably distilled water and alcohol are added in the same ratio. As the organic solvent, any one or more of low gold alcohol having 1 to 4 carbon atoms, propylene glycol, ethyl acetate, butylene glycol, glycerin, ethyl acetate, hexane, and methylene chloride may be used.

In addition, the mixing ratio of the lactic acid bacteria fermentation supernatant of the third step and the extraction solvent is preferably 50:50 to 25:75 by weight.

The fermented herbal extract prepared according to the present invention can be used as a functional cosmetic composition for whitening, wrinkle improvement, UV protection, anti-atopic and athlete's foot prevention or improvement.

The cosmetic composition of the present invention may be prepared in any formulation conventionally prepared in the art, for example, a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, It may be formulated as oil, powder foundation, emulsion foundation, wax foundation and spray, but is not limited thereto. More specifically, it may be prepared in the form of a flexible lotion, astringent lotion, nourishing lotion, nourishing cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder.

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component. can

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and in the case of a spray, chlorofluorohydrocarbon, propane/butane or a propellant such as dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as a carrier component.

When the formulation of the present invention is a suspension, water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as polyoxyethylene sorbitol ester, microcrystalline cellulose, etc. may be used as a carrier component.

When the formulation of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, or the like may be used as a carrier component.

The present invention discloses a whitening, anti-wrinkle, UV protection, anti-atopic and athlete's foot prevention or improvement functional cosmetic composition containing a fermented herbal extract. Hereinafter, the present invention will be described in more detail by way of Examples. However, the following examples are only for illustrating the present invention and are not intended to limit the scope of the present invention.

For the herbs of the fermented herbal extract of the present invention, rosemary, lavender, sage, basil, peppermint, chamomile, oregano, lemon balm, and stevia collected at Sangsang Herb Brand Farm near Cheongju were dried and pulverized, respectively, and used as a dry powder. Lactobacillus plantarum (L. plantarum, KFCC-11028) was purchased from the Korea Microbial Conservation Center and used as lactic acid bacteria, and MRS culture medium (Difco ^TM MRS) was purchased from BD.

[Example 1] Preparation of fermented herbal extract

900 g of distilled water equal to 900 g of rosemary powder was placed in a fermenter, sterilized at 100° C. for 5 minutes, and then left at 37° C. Then, 200 mL (1 X 10 ⁸ cells) of Lactobacillus plantarum (KFCC-11028), which was separately cultured in liquid phase in MRS medium, was added to the fermenter, and the temperature was 37° C., pH 7, and the stirring speed was 100 rpm. It was fermented for 3 days under the fermentation conditions of After centrifuging the fermented product obtained in the above step, the precipitate and cells were removed to obtain a fermentation broth. 900 g of distilled water was added to the fermentation broth obtained in the above step, put it in an extractor equipped with a cooling condenser, and heated to a temperature of 80° C. for 6 hours, and then 800 L of distilled liquid was recovered. The fermented extract obtained above was first filtered with a 300 mesh filter paper, left at room temperature for 1 week, and then filtered twice with 150 mm filter paper to prepare a fermented rosemary extract of the present invention.

In the same manner as the above method, fermented lavender extract, fermented sage extract, fermented basil extract, fermented mint extract, fermented chamomile extract, fermented oregano extract, fermented lemon balm extract, and fermented stevia extract were respectively prepared.

[Example 2] Preparation of mixed fermented herbal extract

Unlike Example 1, the mixed fermented herbal extract was prepared as follows. Weigh and mix 100 g of rosemary powder, 100 g of lavender powder, 100 g of sage powder, 100 g of basil powder, 100 g of peppermint powder, 100 g of chamomile powder, 100 g of oregano powder, 100 g of lemon balm powder and 100 g of stevia powder. Using 900 g of one herbal mixed powder, 1,800 mL of lactic acid bacteria fermented herbal mixed extract was prepared. A specific manufacturing method is the same as in Example 1.

[Comparative Example 1] Preparation of mixed herbal extract

Herbal mixture of 100 g of rosemary powder, 100 g of lavender powder, 100 g of sage powder, 100 g of basil powder, 100 g of peppermint powder, 100 g of chamomile powder, 100 g of oregano powder, 100 g of lemon balm powder and 100 g of stevia powder. 900 g of distilled water equal to 900 g of powder was placed in an extractor equipped with a cooling condenser, heated at a temperature of 80° C. for 6 hours, and the distilled solution was recovered to obtain an herbal extract.

[Preparation Example 1] Preparation of cosmetic composition of mixed fermented herbal extract

After preparing the mixed fermented herbal extract prepared in Example 2, adjusting the temperature of the extract to 70°C, heating the emulsifier at 70°C to obtain an emulsifier in a solution state, the mixed fermented herbal extract and the emulsifier, and moisturizing oil , a moisturizer and purified water were mixed and stirred while maintaining 70 °C. Thereafter, it was cooled slowly to prepare a lotion-type cosmetic composition. Olive oil was used as the emulsifier used in the preparation, olive oil was used as the moisturizing oil, and glycerin was used as the moisturizing agent to prepare a cosmetic composition in the composition ratio as shown in Table 1.

Cosmetic composition mixing ratio Raw material Content (% by weight) Mixed Fermented Herb Extract (Example 2) 3-5 emulsifier 2-3 moisturizing oil 6-7 moisturizer 9-10 Purified water 75-80 Sum 100

[Comparative Example 2] Preparation of cosmetic composition of mixed herbal extract

Example 2 A cosmetic composition of a mixed herbal extract was prepared in the same manner as in the above method using the mixed herbal extract of Comparative Example 1 instead of the mixed fermented herbal extract.

[Experimental Example 1] Tyrosinase inhibitory effect measurement

Tyrosinase (tyrosinase) inhibitory effect was measured using the dopachrome method. After mixing 150 μl of mushroom tyrosinase-150 unit, 225 μl of 2.5 mM L-tyrosine, 225 μl of 0.4 M hepes buffer (pH 6.8), ethanol solution or 300 μl of fermented herb extract with a concentration of 1 mg/mL, before incubation, 15 minutes Each absorbance after incubation was measured at a wavelength of 475 nm. In order to examine the tyrosinase inhibitory effect according to the presence or absence of lactic acid bacteria fermentation, the herbal extract prepared in Comparative Example 1 was also carried out in the same manner as described above. As a positive control, kojic acid was used, and the absorbance of each of the measured sample solutions was calculated, and the tyrosinase activity inhibition level was calculated using the following formula.

Tyrosinase Inhibition (%) = (D - C) - (B - A) / (D - C) × 100

A and B are the absorbances before and after incubation of the solution to which the sample is added, respectively, and C and D are the absorbances before and after incubation of the solution without the sample, respectively. When these tyrosinase inhibitory effects are expressed as 100, it means complete inhibition, and when 0, it means no inhibition at all.

Experimental results As shown in Table 2, the tyrosinase activity inhibitory effect of various fermented herbal extracts at 0.5 mg/mL concentration was in the order of mixed herbs (Example 2), lavender, oregano, mint, sage, rosemary, lemon balm, chamomile, stevia, and basil. 78.7, 65.7, 65.1, 64.6, 63.8, 62.5, 61.5, 59.2, 57.1, and 54.1% of tyrosinase inhibitory activity were exhibited. The mixed fermented herbal extract had the highest tyrosinase activity inhibitory activity, and the fermented basil extract had the lowest. As a result of comparison using kojic acid, which is a representative substance showing the inhibitory effect of tyrosinase enzyme, as a positive control, kojic acid was 69.8% under the same conditions, and it was confirmed that the mixed fermented herbal extract was 8.9% higher. The activity inhibitory effect was measured to be 11.6% higher in the herbal extract fermented with lactic acid bacteria. Therefore, it was found that fermenting herbs with lactic acid bacteria had the highest tyrosinase activity inhibition rate and thus had a whitening effect.

Measurement result of tyrosinase inhibitory effect sample Tyrosinase activity inhibition rate (%) Control (kojic acid) 69.8 Example 1 Fermented Rosemary Extract 62.5 Fermented Lavender Extract 65.7 Fermented Sage Extract 63.8 Fermented Basil Extract 54.1 Fermented Mint Extract 64.6 Fermented Chamomile Extract 59.2 Fermented Oregano Extract 65.1 Fermented Lemon Balm Extract 61.5 Fermented Stevia Extract 57.1 Example 2 (mixed fermented herbal extract) 78.7 Comparative Example 1 (mixed herbal extract) 67.1

[Experimental Example 2] Measurement of melanin production

To measure the amount of melanin production, melanoma cell B16F10 was treated with each sample and positive control arbutin at concentrations of 2.5%, 5.0%, and 10.0% (v/v) to measure the whitening activity. Melanoma cell B16F10 was inoculated in a 96-well plate at a concentration of 1.5 × 10 ⁴ cells/well, and the sample was treated for 3 days. After 3 days, after removing the medium from each well, it was washed with PBS, 1 mL of 1 N NaOH was added to each well, and then stirred so that the melanin component was eluted. To measure the content of eluted melanin, the absorbance was measured at 400 nm using a microplate reader. The herbal extract prepared in Comparative Example 1 was also carried out in the same manner as above in order to determine the amount of melanin production according to the presence or absence of fermentation of lactic acid bacteria. As a positive control, hydroquinone, known as a representative whitening agent, was used.

As a result of the experiment, as shown in Table 3, the amount of melanin production of various fermented herbal extracts was in the order of mixed herb (Example 2), oregano, lavender, mint, sage, rosemary, lemon balm, chamomile, stevia, and basil at a concentration of 0.1 mg/mL. 47.6, 52.8, 53.8, 55.7, 58.6, 61.3, 62.8, 64.4, 66.5% of melanin production was shown, and the mixed fermented herbal extract had the lowest melanogenesis performance and the fermented basil extract had the highest. The amount of melanin production according to the presence or absence of lactic acid bacteria fermentation was measured to be 11.3% lower in the herbal extract fermented with lactic acid bacteria.

Measurement result of melanin production sample Melanin production (%) Control (hrdroquinone) 40.1 Example 1 Fermented Rosemary Extract 58.8 Fermented Lavender Extract 53.8 Fermented Sage Extract 58.6 Fermented Basil Extract 66.5 Fermented Mint Extract 55.7 Fermented Chamomile Extract 62.8 Fermented Oregano Extract 52.8 Fermented Lemon Balm Extract 61.3 Fermented Stevia Extract 64.4 Example 2 (mixed fermented herbal extract) 47.6 Comparative Example 1 (mixed herbal extract) 58.3

[Experimental Example 3] Measurement of anti-wrinkle effect by collagenase activator

One of the causes of skin wrinkles is a deficiency of dermal collagen, that is, collagen. Collagen is a major protein constituting the dermis of the skin and plays a role in maintaining the structure and elasticity of the skin. It is known that collagen production decreases with age, and the degree of decomposition increases to induce depression of the dermal layer of the skin to generate wrinkles of the skin. In addition, matrix metalloproteinase (MMP), such as collagenase, is expressed by ultraviolet light, which is a cause of photoaging, thereby increasing the degradation of collagen, thereby generating wrinkles. Therefore, by checking whether the fermented herbal extract of the present invention inhibits collagenase activity, it was attempted to see the effect of improving skin wrinkles.

To 0.8 µL of 0.05 M tricine buffer (pH7.5) as a reaction group, 0.05 µL of substrate solution and 0.05 µL of sample solution in which FALGPA at a concentration of 0.2 mM were dissolved, and 0.05 mL of collagenase (3 mg/mL) was added and left at 20°C for 18 hours. Thereafter, 10 μL of 1 N acetate was added to stop the reaction, and the inhibitory activity was measured at an absorbance of 324 nm. The herbal extract prepared in Comparative Example 1 was also carried out in the same manner as above in order to determine the inhibition rate of collagenase activity according to the presence or absence of lactic acid bacteria fermentation. As a positive control group, Retinol (vitamin A), a representative anti-wrinkle substance, was used.

Experimental results As shown in Table 4, at a concentration of 1 mg/mL of various fermented herbal extracts, the inhibition rate of circulating collagenase activity was 45.9 ~ 66.7%. % was measured as high. It was confirmed that the collagenase activity inhibition rate of the lactic acid bacteria fermented mixed herbal extract was high, and the skin wrinkle improvement effect was confirmed.

Collagenase activity inhibitory effect measurement result sample Collagenase activity inhibition rate (%) Control (Retinol) 82.3 Example 1 Fermented Rosemary Extract 50.1 Fermented Lavender Extract 60.4 Fermented Sage Extract 54.5 Fermented Basil Extract 49.6 Fermented Mint Extract 52.1 Fermented Chamomile Extract 53.2 Fermented Oregano Extract 49.8 Fermented Lemon Balm Extract 48.1 Fermented Stevia Extract 45.9 Example 2 (mixed fermented herbal extract) 66.7 Comparative Example 1 (mixed herbal extract) 55.9

[Experimental Example 4] Measurement of UV protection effect

The UV-blocking effect was tested by measuring the inhibition of MMP-1 expression using UVA treatment (ELISA method).

CCD-986sk was cultured at a concentration of 1.5 × 10 ⁵ cells/mL in a 35 mm dish until it reached about 80% confluency. After removing the original medium before UV irradiation, it was washed with PBS to remove the serum component in the medium, and then UVA (6.3 J/cm2) was irradiated for 24 hours using a filter in UV light (Coralife, 35W, UV). After UVA irradiation, the culture medium was cultured for 24 hours by administering the Angelica asiatica extract at a concentration of 1 mg/mL in DMEM medium without FBS.

MMP-1 expression level induced by UVA irradiation was measured using the method used by Dunsmore et al. (Dunsmore et al., 1996). After UVA irradiation, the sample was treated and cultured for 24 hours was dispensed into a 96 well plate and reacted at 4°C overnight. After washing 3 times with TBS (phosphate buffered saline + 0.05% Tween 20) and blocking with 3% PBS at 37°C for 1 hour, using a primary antibody (monoclonal anti-MMP-1 antibody) as a blocking buffer 1: It was diluted to 3,000 and reacted at 37°C for 90 minutes. After washing with PBS, the secondary antibody (alkaline phosphatase conjugated Goat anti-mouse IgG) was diluted 1:3,000 in blocking buffer, reacted at 37°C for 90 minutes, washed with PBS, and then alkaline phosphatase substrate solution (1 mg/mL, p-mitrophenyl phosphate in diethanolamine buffer) was added and reacted at room temperature for 30 minutes. After completely stopping the reaction with 3 N NaOH, the absorbance was measured at 405 nm using a microplate reader. Ascorbic acid (vitamin C) was used as a positive control.

As shown in Table 5, ascorbic acid, a positive control, showed the most suppressed expression rate at 1 mg/mL concentration as shown in Table 5, and Example 2 (mixed fermented herbal extract) extract showed 123.2% close to ascorbic acid in the sample. It showed the lowest expression rate. Other samples also showed a value of 132.1% or less, indicating a relatively low expression rate, indicating that there was an inhibitory effect on MMP-1 expression on UVA.

UV protection effect measurement result sample UV protection rate (%) Control (ascorbic acid) 121.1 Example 1 Fermented Rosemary Extract 130.5 Fermented Lavender Extract 129.1 Fermented Sage Extract 128.4 Fermented Basil Extract 125.6 Fermented Mint Extract 129.9 Fermented Chamomile Extract 127.4 Fermented Oregano Extract 130.1 Fermented Lemon Balm Extract 132.1 Fermented Stevia Extract 131.8 Example 2 (mixed fermented herbal extract) 123.2 Comparative Example 1 (mixed herbal extract) 129.4

[Experimental Example 5] Sensory evaluation of alleviation and improvement of atopic symptoms

In order to evaluate the anti-atopic effect of the cosmetic composition prepared in Preparation Example 1 and the cosmetic composition prepared in Comparative Example 2, it was investigated and compared as follows.

Sensory evaluation was performed on the degree of alleviation and improvement of atopic symptoms, moisturizing power, and preference for the prepared cosmetic composition. At this time, the test subject was evaluated by applying it to the skin showing atopic symptoms or dry symptoms at least once a day for a week, targeting 10 adults with atopic symptoms over 20 years of age. The evaluation score ranged from 1 to 5, and the best score was 5, and the results are shown in Table 5 below (1: very bad, 2: bad, 3: average, 4: good, 5: very good). ).

As shown in Table 6, as a result of the sensory evaluation, it was evaluated that the cosmetic composition of the present invention exhibits a more excellent effect than that of Comparative Example 2 in the degree of improvement of atopic symptoms. In particular, it was evaluated that erythema and itch symptoms were greatly alleviated, and in terms of moisturizing power, it was evaluated as showing excellent moisturizing power, and it was evaluated that the cosmetic composition of Preparation Example 1 had a higher preference than the cosmetic composition of Comparative Example 2.

That is, in the case of the cosmetic composition of the present invention, the degree of alleviation and improvement of atopic symptoms has not been medically proven, but it was determined that it could indirectly exhibit the alleviation effect of atopic symptoms through the skin soothing effect according to excellent skin moisturizing and antibacterial properties. .

Sensory evaluation results for the alleviation and improvement of atopic symptoms Preparation Example 1 Comparative Example 2 Relief of atopic symptoms 4.5 3.1 moisturizing power 4 3.3 preference 4.5 3.1

[Experimental Example 6] Confirmation of antibacterial activity

In order to confirm the antibacterial activity of the fermented herbal extract of the present invention against athlete's foot bacteria, representative athlete's foot bacteria Trichophyton rubrum (KCTC 6345), Trichophyton mentagrophytes (KCTC 6077), microsporum car Four species of Nice (Microsporum canis, KCTC 6348) and Candida albicans (KCTC 7965) were distributed from a microbial resource center and their growth inhibitory activity was tested by the paper disc diffusion method.

40 g of dextrose and 10 g of peptone were mixed, distilled water was added to make a total of 1 L, and then sterilized to prepare SDB (Saboraud dextrose broth) medium, and 15 g of agar was added to the same composition. By adding (40 g of dextrose, 10 g of peptone, 15 g of agar, and 1 L of distilled water in total), SDA (Saboraud dextrose agar) was prepared. After that, the received strain was inoculated into the SDB medium and aerobically cultured at 28°C for 7 days (however, Candida albicans cultured for 1 day), 1 mL of each culture solution and 2 mL of soft agar (0.7%) It was mixed evenly and spread on SDA medium. After inoculation on the SDA medium, 50 μL of each of the extracts of Examples 1, 2 and Comparative Example 1 were placed on paper discs inoculated with each bacteria and aerobically cultured at 28°C for 14 days ( However, Candida albicans cultured for 1 day). Antifungal activity was confirmed by looking at the size of the growth inhibition ring (clear zone) formed around the filter paper, and the results are shown in Table 7 below. At this time, it was judged that there was antifungal activity when the size of the growth inhibitory ring was 10 mm or more due to each extract.

Experimental results As shown in Table 7, the mixed fermented herbal extract of Example 2 of the present invention inhibited growth of Trichophyton rubrum, Trichophyton mentagrophytes, Microsporum canis, and Candida albicans compared to other extracts. With a size of 30 mm or more, it was found that the antifungal properties were remarkably excellent, and it was judged that there was an antibacterial effect of athlete's foot bacteria.

Measurement result of antibacterial activity sample
(50 μL) Growth inhibition ring (clear zone (mm)) Trichophyton rubrum Trichophyton
Mentagrophytes microforum canis candida
albicans Example 1 Fermented Rosemary Extract +++ +++ ++ +++ Fermented Lavender Extract ++ ++ +++ ++ Fermented Sage Extract +++ ++ +++ +++ Fermented Basil Extract + + + + Fermented Mint Extract ++++ +++ +++ +++ Fermented Chamomile Extract +++ +++ +++ +++ Fermented Oregano Extract ++++ ++++ ++++ ++++ Fermented Lemon Balm Extract +++ +++ +++ +++ Fermented Stevia Extract + + ++ ++ Example 2
(Mixed Fermented Herb Extract) +++++ +++++ +++++ +++++ Comparative Example 1 (mixed herbal extract) +++ +++ ++++ +++++ 10mm or less: +, 11~16mm: ++, 17~22mm: +++, 23~29mm: ++++ 30mm or more: +++++

The present invention has an excellent effect of providing anti-atopic and athlete's foot prevention or improvement functional cosmetics or quasi-drugs, in addition to whitening, anti-wrinkle and UV-blocking functional cosmetics containing lactic acid bacteria fermented herbal extract, and in particular, cosmetics in formulations such as creams, lotions, and ointments Alternatively, it has an excellent effect that it can be used as a new material for quasi-drugs. Therefore, by providing a cosmetic composition containing useful ingredients contained in the lactic acid bacteria fermented herbal mixed extract of the present invention, there is an excellent effect of improving whitening, wrinkle improvement, UV protection, anti-atopic and athlete's foot prevention or improvement functions. It is a very useful invention.

Claims (5)

(a) 4 or more herbs selected from herbs consisting of rosemary, lavender, sage, basil, mint, chamomile, oregano, lemon balm, and stevia are mixed in the same ratio to dry powder, and then the same amount is 1:1 (w/w ) in a fermenter with distilled water, high-temperature sterilization at 100°C for 5 minutes, and cooling to 37°C to prepare fermentation raw materials;
(b) inoculating the fermented raw material obtained in the above step with vegetable lactic acid bacteria and fermenting for 3 to 5 days at 35 to 37° C. and pH 7.0 to obtain a fermented product;
(c) centrifuging the fermented product obtained in the above step to obtain a fermentation supernatant;
(d) the fermentation supernatant obtained in the above step is extracted with hot water at 80° C. for 6 hours with an extraction solvent in which alcohol is added with an equal amount of 1:1 (w/w) distilled water to recover the distilled liquid, and then the remaining fermented herbs obtaining an extract;
(E) atopy prevention and improvement functional lactic acid bacteria fermented herbal mixed extract prepared including the step of filtering the remaining fermented herbal extract in the step with a filter paper. delete delete delete delete

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