KR102675543B1 - Composition comprising fermented snowberry and its use for improving skin condition - Google Patents

Abstract

It relates to a composition comprising fermented snowberry and its use for improving skin condition. A composition containing fermented snowberry according to one aspect can be useful for improving skin-related conditions, such as preventing skin aging, improving skin wrinkles, enhancing skin elasticity, whitening skin, moisturizing skin, and strengthening skin barrier.

Description

Composition comprising fermented snowberry and its use for improving skin condition {Composition comprising fermented snowberry and its use for improving skin condition}

It relates to cosmetic compositions, food compositions containing fermented snowberries, and their use for improving skin condition.

The skin is the human body's primary defense shield from the external environment and is one of the important organs of the human body that regulates body temperature, barrier function, and various physiological functions. Human skin goes through an aging process as we age, and the causes can be broadly divided into intrinsic aging and extrinsic aging. Intrinsic aging is a process that no one can avoid as we age. Wrinkles appear, skin loses elasticity, moisture within skin cells is lost, and the structure of the stratum corneum changes. On the other hand, extrinsic aging is caused by external stress such as environmental factors, and the main cause is ultraviolet rays, so it is also called photoaging.

Matrix proteins such as collagen and elastin exist in the skin, and when these matrix proteins are reduced by decomposition enzymes such as MMP (matrix metalloproteinase), wrinkles occur in the skin and elasticity decreases. Conversely, increasing the synthesis of procollagen in the skin helps improve skin wrinkles and increase skin elasticity.

Another phenomenon of skin aging is that the ability to retain moisture decreases, causing the skin to become dry and rough. Water makes up about 70% of the human body and helps keep the skin moist and shiny. The stratum corneum of a healthy epidermis contains 15 to 20% moisture, and if this level falls below 10%, the skin barrier will malfunction, the skin will dry out, and wrinkles will increase. The protein that regulates water movement includes aquaporin (AQP). Aquaporin-3 (AQP-3), which is mainly present in the keratinocytes of the epidermis and is a moisture channel through which water, glycerol, and urea move, provides smooth moisture between cells. Additionally, an increase in filaggrin, a moisturizing ingredient that forms the skin barrier, can not only increase skin moisturization but also strengthen the skin barrier.

In addition, melanin pigment determines skin color, and melanin produced from excessive stimulation can cause skin problems such as spots, freckles, and hyperpigmentation.

Meanwhile, Patent Document 1 discloses a composition containing snowberry extract. However, there is no research on the efficacy of fermented snowberry products.

Accordingly, the present inventor completed the present invention by confirming the use of fermented snowberry for improving skin condition.

KR 10-1601908 B1

A composition comprising a fermented product obtained by fermenting snowberries with microorganisms is provided.

A method for producing snowberry fermentation product is provided.

A method of improving the skin condition of a subject is provided, comprising administering an effective amount of snowberry ferment or a composition containing the same to a subject in need thereof.

Provided is the use of snowberry fermentation for use in preparing compositions for improving skin condition.

One aspect provides a composition comprising a fermented product obtained by microbially fermenting snowberries.

The term "Snowberry" refers to a plant of the genus Symphoricarpos sp. , and is also called snowberry, waxberry, and ghostberry in English. Depending on the type, snowberry berries come in different colors, including white (e.g. common snowberry), pink (e.g. little leaf snowberry), red (e.g. coral berry), and blackish purple (e.g. Chinese coral berry). represents.

Exemplary types of snowberries include Common snowberry ( Symphoricarpos albus ), Mexican snowberry ( Symphoricarpos mexicanus ), Little leaf snowberry ( Symphoricarpos microphyllus ), and Creeping snowberry ( Symphoricarpos ). mollis ), Western snowberry or Wolfberry, Symphoricarpos occidentalis , Coralberry, Symphoricarpos orbiculatus , Mountain snowberry, Symphoricarpos oreophilus , Roundleaf snowberry, Symphoricarpos rotundifolius , Chinese coralberry ( Symphoricarpos sinensis ), etc.

In one embodiment, the snowberry may be common snowberry, Mexican snowberry, little leaf snowberry, creeping snowberry, western snowberry, coral berry, mountain snowberry, Chinese coral berry, or a combination of two or more thereof. .

The fermented product can be obtained by fermenting snowberries using microorganisms. The above “fermented product obtained by fermenting snowberries with microorganisms” is used interchangeably with “snowberry fermented product.” Snowberries are known to be consumed only by animals because they contain ingredients that cause mild vomiting when consumed by humans. However, since the toxicity of snowberries is removed through fermentation, the fermented snowberries can also be consumed by humans as food.

The microorganism may be one or more types of microorganisms selected from the group consisting of Bacillus sp. , Lactobacillus sp. , and Leuconostoc sp. , but is not limited thereto.

The Bacillus sp. microorganisms include all aerobic or facultative anaerobic bacilli widely distributed in nature. The microorganisms of the Bacillus genus are not limited to a specific type, but for example, Bacillus subtilis , B. coagulans , B. velezensis , Bacillus licheniphor It may be selected from the group consisting of B. licheniformis , B. polyfermenticus , B. methylotrophicus , and B. amyloliquefaciens .

The Lactobacillus genus ( Lactobacillus sp. ) microorganisms are not limited to a specific type, but for example, Lactobacillus acidophilus ( L. acidophilus ), Lactobacillus casei ( L. casei ), Lactobacillus gasseri ( L gasseri ), Lactobacillus delbrueckii ssp. bulgaricus , Lactobacillus helveticus ( L. helveticus ), Lactobacillus fermentum ( L. fermentum ), Lactobacillus paracasei ( L. paracasei ) ), Lactobacillus plantarum ( L. plantarumm ), Lactobacillus reuteri ( L. reuteri ), Lactobacillus rhamnosus ( L. rhamnosus ), and Lactobacillus salivarius ( L. salivarius ). there is.

The microorganisms of the Leuconostoc sp. include Leuconostoc mesenteroides ( L. mesenteroides ), Leuconostoc citreum ( L. citreum ), Leuconostoc holzapfelii ( L. holzapfelii ), and Leuconostoc It may be selected from the group consisting of L. lactis.

The fermentation conditions can be selected at an appropriate temperature and time depending on the type of microorganism. For example, the fermentation may be performed at 20 to 40° C., but is not particularly limited thereto. In addition, the fermentation may be performed for 1 to 200 hours, 1 to 180 hours, 24 to 200 hours, 24 to 180 hours, 24 to 120 hours, 48 to 200 hours, 48 to 180 hours, or 48 to 120 hours. , but is not particularly limited thereto.

The fermentation medium can be appropriately selected depending on the type of microorganism. Depending on the type of microorganism, the medium may be a known medium or a conventional medium capable of cultivating the microorganism. For example, the medium may include one or a mixture of two or more selected from glucose, yeast extract, malt extract, peptone, glutamic acid, K ₂ HPO ₄ , KH ₂ PO ₄ , and NaCl, but is not limited thereto. .

In one embodiment, when the microorganism is a Bacillus genus microorganism, the medium may include glucose, peptone, yeast extract, glutamic acid, K ₂ HPO ₄ , and KH ₂ PO ₄ .

In another embodiment, when the microorganism is a microorganism of the Lactobacillus genus, the medium may include glucose, peptone, and yeast extract.

In another embodiment, when the microorganism is a microorganism of the Leuconostoc genus, the medium may include glucose, peptone, yeast extract, K ₂ HPO ₄ , and NaCl.

The fermented product may include a fermented broth obtained by fermenting snowberries with microorganisms, a diluted or concentrated liquid of the fermented broth, a dried product obtained by drying the fermented broth, or these crude or purified products, fractions thereof, etc.

The fermented product is 0.0001 to 99.9999% by weight, 0.0001 to 90% by weight, 0.0001 to 50% by weight, 0.0001 to 10% by weight, 0.001 to 99.9999% by weight, 0.001 to 90% by weight, 0.001 to 50% by weight, based on the total weight of the composition. , 0.001 to 10% by weight, 0.01 to 99.9999% by weight, 0.01 to 90% by weight, 0.01 to 50% by weight, 0.01 to 20% by weight, or 0.01 to 10% by weight, but is not particularly limited thereto.

The term “included as an active ingredient” means that the fermented product of the present specification is added to an extent that can produce the effects mentioned above. Additionally, this may include formulation in various forms by adding various ingredients as sub-ingredients for drug delivery and stabilization.

The composition may be a composition for improving skin condition, improving skin beauty, or preventing or treating skin diseases.

In one embodiment, the composition may be used to improve skin condition.

The skin condition improvement may be preventing skin aging, improving skin wrinkles, improving skin elasticity, whitening skin, moisturizing skin, strengthening skin barrier, or a combination of two or more thereof.

The term “skin aging” refers to both tangible and intangible changes that occur in the skin as one ages, such as thinning of the epidermis, number of cells or blood vessels in the dermis, DNA damage repair ability, cell replacement cycle, wound recovery, It refers to a decrease in skin barrier function, epidermal moisture retention, sweat secretion, sebum secretion, vitamin D production, physical damage defense, chemical removal ability, immune response, sensory function, and temperature regulation.

The fermented product may be used for preventing skin aging caused by exogenous or endogenous factors, for improving skin aging, or for anti-aging. The exogenous factors refer to various external factors, such as ultraviolet rays (light). The endogenous factors are also referred to as chronological factors and mainly refer to factors that occur due to the passage of time. In other words, the skin aging is not only a premature aging phenomenon induced by external stimuli such as ultraviolet rays, pollution, cigarette smoke, chemicals, etc., but also a natural aging phenomenon that occurs as the proliferation of skin cells decreases with age. , that is, it includes skin aging due to age, and is a concept that includes wrinkles, loss of elasticity, skin sagging, and dryness. In addition, wrinkles include stimulation caused by changes in internal and external factors that change the components that make up skin tissue, causing wrinkles.

The aging may be photoaging. The term “photoaging” is a phenomenon caused by external environmental factors, the most representative factor being ultraviolet rays. Ultraviolet rays cause damage to biological components such as activation of proteolytic enzymes, chain cutting of matrix proteins, and abnormal cross-linking, and repetition of this mechanism causes skin aging that is evident in appearance. Therefore, the composition may be used to prevent, improve, or treat photoaging caused by ultraviolet rays (ultraviolet, UV).

The term “skin wrinkle improvement” is interpreted to mean not only the action of reducing the number or depth of wrinkles on the skin, but also includes suppressing skin aging through wrinkle improvement.

The term “promoting skin elasticity” may refer to any action that increases skin elasticity that has been reduced due to causes such as aging or prevents a decrease in skin elasticity.

The term “skin whitening” may mean not only brightening skin tone by inhibiting the synthesis of melanin pigment, but also improving skin hyperpigmentation such as spots or freckles caused by ultraviolet rays, hormones, or genetics.

The term “skin moisturizing” can refer to any action that maintains skin moisture or prevents moisture loss. The skin moisturizing effect can help improve skin wrinkles and increase elasticity.

The term “skin barrier strengthening” can refer to any action that improves the function of the skin barrier, which is located on the outermost layer of the skin and prevents moisture and nutrition loss.

The term “skin disease” may be a disease caused by skin aging, skin hyperpigmentation, or impaired skin barrier function. The term “prevention” includes suppressing the occurrence of a disease. The term “treatment” includes inhibiting, alleviating, or eliminating the development of a disease.

Details regarding skin aging are as described above. Therefore, the skin aging may be photoaging or skin aging due to age.

The skin hyperpigmentation may include spots or freckles caused by ultraviolet rays, hormones, or genetics.

The damage to the skin barrier function may mean any change that appears in the skin due to decreased or damaged skin barrier function. For example, it may include increased skin wrinkles, dryness, dermatitis, atopic dermatitis, allergic dermatitis, acne, etc.

The composition can inhibit the expression of MMP-1. As the skin ages or due to irritants such as external ultraviolet rays and fine dust, it secretes various factors. Among these, it secretes MMP (matrix metalloproteinase), an enzyme that decomposes extracellular matrix proteins, to break down matrix proteins such as collagen and elastin fibers. Decompose. In particular, collagen accounts for most of the skin's matrix structure, and when it is reduced by decomposition enzymes such as MMPs, wrinkles occur in the skin and elasticity decreases. The composition may exhibit an anti-aging effect on the skin by inhibiting the expression of MMP-1.

The composition can increase the expression of procollagen. It is generally known that collagen, which is associated with wrinkles in skin cells, is synthesized in the form of a precursor substance called procollagen. Therefore, by measuring the amount of procollagen peptides, the degree of collagen synthesis within the cell can be determined. The composition can improve skin wrinkles and improve skin elasticity by increasing the expression of procollagen.

The composition can inhibit melanin synthesis. Melanin pigment is synthesized in melanocytes in the basal layer of the skin. Melanin synthesized by stimuli such as ultraviolet rays accumulates in melanosomes and is then transferred back to adjacent keratinocytes, giving the stratum corneum a reddish-brown color. Melanin determines eye, hair, and skin color and is mainly responsible for scavenging free radicals generated by ultraviolet rays and protecting the skin from ultraviolet rays. However, melanin produced due to excessive stimulation causes skin problems such as blemishes, freckles, hyperpigmentation, etc., and in severe cases, cell death and skin cancer may occur. The composition can exhibit a skin whitening effect by inhibiting melanin synthesis.

The composition can increase the expression of aquaporin 3 (AQP3). The composition can exhibit a skin moisturizing effect by smoothly supplying moisture to skin cells by increasing the expression of AQP3.

The composition can increase the expression of filaggrin. The filaggrin is a moisturizing ingredient that forms the skin barrier. The composition can exhibit skin moisturizing and skin barrier strengthening effects by increasing the expression of filaggrin.

Accordingly, the composition may inhibit the expression of MMP-1, inhibit melanin synthesis, or increase the expression of any one or more of procollagen, AQP-3, and filaggrin.

In one example, the fermented product was confirmed to be excellent in preventing skin aging, improving skin wrinkles, improving skin elasticity, whitening skin, moisturizing skin, and strengthening skin barrier. In addition, when using microorganisms of the genus Bacillus, Lactobacillus, and Leuconostoc as fermentation microorganisms, the effect was all excellent, but in particular, when microorganisms of the genus Bacillus were used, the best effect was shown.

The composition may be a cosmetic composition.

The cosmetic composition can be manufactured in any commonly manufactured formulation. For example, the cosmetic composition includes lotion, cream, essence, cleansing foam, cleansing water, pack, ampoule, body lotion, body oil, body gel, shampoo, rinse, hair conditioner, hair gel, foundation, lipstick, mascara, and makeup. It may have a base or skin-adhesive cosmetic formulation. The skin-adhesive cosmetic formulation may be, for example, a mask pack, but is not limited thereto.

Ingredients included in the cosmetic composition may include ingredients commonly used in cosmetic compositions in addition to the composition as an active ingredient, for example, conventional auxiliaries and carriers such as stabilizers, solubilizers, vitamins, pigments, and fragrances. may include.

Additionally, the composition may be a composition for external skin application.

The external skin preparation may be a cream, gel, ointment, skin emulsifier, skin suspension, transdermal delivery patch, drug-containing bandage, lotion, or a combination thereof. The skin external preparations include ingredients commonly used in skin external preparations such as cosmetics and medicines, such as aqueous ingredients, oil-based ingredients, powder ingredients, alcohols, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, and fragrances. , colorants, various skin nutrients, or a combination thereof may be appropriately mixed according to need. The skin external preparations include metal sequestrants such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, and gluconic acid, caffeine, tannin, belafamil, licorice extract, glablidin, and calin. Hot water extract of fruit, various herbal medicines, drugs such as tocopherol acetate, glytylitinic acid, tranexamic acid and its derivatives or salts, vitamin C, magnesium ascorbate phosphate, ascorbate glucoside, arbutin, kojic acid, glucose, fructose, Sugars such as trehalose can also be appropriately mixed.

The composition may be a food composition. The composition may be a health functional food composition.

The food composition may be used alone or in combination with other foods or food ingredients, and may be used appropriately according to conventional methods. The mixing amount of the active ingredient can be appropriately determined depending on the purpose of use (prevention, health, or therapeutic treatment). There are no particular restrictions on the types of health functional foods. Among the types of health functional foods, beverage compositions may contain various flavoring agents or natural carbohydrates as additional ingredients like ordinary beverages. The natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As a sweetener, natural sweeteners such as thaumatin and stevia extract or synthetic sweeteners such as saccharin and aspartame can be used. The health food composition also contains nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, and carbonated beverages. It may contain the carbonating agent used, or a combination thereof. The health functional food composition may also contain pulp for the production of natural fruit juice, fruit juice beverage, vegetable beverage, or a combination thereof.

The composition may be a pharmaceutical composition.

In one embodiment, the pharmaceutical composition may be used to prevent or treat skin aging.

The skin aging may be photoaging or skin aging due to age.

The pharmaceutical composition may additionally include a pharmaceutically acceptable diluent or carrier. The diluent may be lactose, corn starch, soybean oil, microcrystalline cellulose, or mannitol, and the lubricant may be magnesium stearate, talc, or a combination thereof. The carrier may be an excipient, disintegrant, binder, lubricant, or a combination thereof. The excipient may be microcrystalline cellulose, lactose, low-substituted hydroxycellulose, or a combination thereof. The disintegrant may be calcium carboxymethyl cellulose, sodium starch glycolate, calcium monohydrogen phosphate, or a combination thereof. The binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or a combination thereof. The lubricant may be magnesium stearate, silicon dioxide, talc, or a combination thereof.

The pharmaceutical composition may be formulated as an oral or parenteral dosage form. Oral dosage forms may be granules, powders, solutions, tablets, capsules, dry syrup, or combinations thereof. Parenteral dosage forms may be injectable.

Another aspect provides a method of making a snowberry ferment.

The method includes incubating snowberries with microorganisms to obtain a fermented product.

Obtaining the fermented product includes adding snowberries to the medium and sterilizing them; And it may include the step of inoculating microorganisms in the medium and incubating them to obtain a fermented product.

Details regarding the snowberry, microorganism, and fermented product are as described above.

The snowberry may be added in an amount of 0.01 to 20% by weight, or 0.1 to 10% by weight, based on the total weight of the medium, but is not limited thereto.

The sterilization may be performed using any known conventional method.

The microorganisms can be used in sufficient amounts to allow fermentation to occur, and can be appropriately selected by a person skilled in the art considering the amount of fermentation raw materials, etc.

The incubation temperature and time can be selected at a temperature and time sufficient to allow fermentation to occur, and can be appropriately selected by a person skilled in the art considering the amount of fermentation raw materials, the type of microorganism, etc. For example, the incubation may be performed at 20 to 40° C., but is not particularly limited thereto. Additionally, the incubation may be performed for 1 to 200 hours, 1 to 180 hours, 24 to 200 hours, 24 to 180 hours, 24 to 120 hours, 48 to 200 hours, 48 to 180 hours, or 48 to 120 hours. , but is not particularly limited thereto.

The fermentation can be performed in a medium capable of cultivating microorganisms. Depending on the type of microorganism, the medium may be a known medium or a conventional medium capable of cultivating the microorganism. Exemplary medium components depending on the type of microorganism are as described above.

The method may further include the step of isolating bacteria from the fermented product. Isolation of the bacteria can be done using known conventional methods, for example, standing, centrifugation, and combinations thereof.

The method may further include filtering the fermented product. The filtration may be performed using any known conventional method.

Another aspect provides a method of improving the skin condition of a subject comprising administering to a subject in need thereof an effective amount of snowberry ferment or a composition comprising the same.

Details regarding the snowberry fermentation product, composition, and skin condition improvement are as described above.

The term “effective amount” refers to an amount effective enough to produce the above-mentioned effects.

The terms “administering,” “introducing,” and “implanting” are used interchangeably and refer to an embodiment of the composition by a method or route that results in at least partial localization of the composition to the desired site. It may mean the arrangement of the composition according to.

Administration can be done by methods known in the art. Administration may be administered directly to the subject by any means, such as, for example, intravenous, intramuscular, oral, transdermal, mucosal, intranasal, intratracheal or subcutaneous administration. You can. The administration may be administered systemically or locally. The administration may be application to the skin.

The subject may be a mammal, such as a human, cow, horse, pig, dog, sheep, goat, or cat. The subject may be an individual in need of improving skin condition, for example, preventing skin aging, improving skin wrinkles, enhancing skin elasticity, whitening skin, moisturizing skin, or strengthening the skin barrier.

The dosage can be prescribed in various ways depending on factors such as formulation method, administration method, patient's age, weight, gender, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity, and those skilled in the art will know these factors. Taking these into account, the dosage can be adjusted appropriately. The frequency of administration can be once a day or more than twice within the range of clinically acceptable side effects, and can be administered at one or two or more locations, daily or at intervals of 2 to 5 days. The number of days of administration can be from 1 to 30 days per treatment. If necessary, the same treatment can be repeated after an appropriate period. For animals other than humans, the dosage per kg is the same as for humans, or the above dosage is converted into, for example, the volume ratio (e.g., average value) of the organs (heart, etc.) between the target animal and human. One dose can be administered.

Another aspect provides the use of snowberry fermentation for use in preparing a composition for improving skin condition.

Details regarding the skin condition improvement, composition, and snowberry fermentation product are as described above.

A composition containing fermented snowberry according to one aspect can be useful for improving skin-related conditions, such as preventing skin aging, improving skin wrinkles, enhancing skin elasticity, whitening skin, moisturizing skin, and strengthening skin barrier.

Figure 1 shows the results of analyzing the expression level of MMP-1 in human fibroblasts. Media Only, untreated control; UV, negative control; EGCG, positive control; Snowberry extract, Comparative Example 1; Snowberry Fermentation 1, Example 1; Snowberry Fermentation 2, Example 2; Snowberry Fermentation 3, Example 3.
Figure 2 shows the results of analyzing the procollagen production rate in human fibroblasts. Media Only, negative control group; TGF-β, positive control; Snowberry extract, Comparative Example 1; Snowberry Fermentation 1, Example 1; Snowberry Fermentation 2, Example 2; Snowberry Fermentation 3, Example 3.
Figure 3 shows the results of analyzing melanin content in melanoma cell lines. Media Only, untreated control; α-MSH, negative control; Arbutin, positive control; Snowberry extract, Comparative Example 1; Snowberry Fermentation 1, Example 1; Snowberry Fermentation 2, Example 2; Snowberry Fermentation 3, Example 3.
Figure 4 shows the relative expression level of AQP-3 in human keratinocytes. Media Only, negative control group; Retinoic acid, positive control; Snowberry extract, Comparative Example 1; Snowberry Fermentation 1, Example 1; Snowberry Fermentation 2, Example 2; Snowberry Fermentation 3, Example 3.
Figure 5 shows the relative expression level of filaggrin in human keratinocytes. Media Only, negative control group; Retinoic acid, positive control; Snowberry extract, Comparative Example 1; Snowberry Fermentation 1, Example 1; Snowberry Fermentation 2, Example 2; Snowberry Fermentation 3, Example 3.

Hereinafter, the present invention will be described in more detail through examples. However, these examples are for illustrative purposes only and the scope of the present invention is not limited to these examples.

Examples 1 to 3. Preparation of snowberry fermentation broth

Snowberry fermentation broth was prepared by fermenting snowberries using microorganisms. The specific manufacturing method is as follows.

First, snowberries were prepared by crushing the raw material. Snowberries were added to the prepared medium and sterilized at 121°C for 15 minutes. The medium is preferably selected appropriately depending on the type of microorganism to be inoculated. For example, a medium prepared with one or a mixture of two or more selected from glucose, yeast extract, malt extract, peptone, glutamic acid, K ₂ HPO ₄ , KH ₂ PO ₄ , and NaCl can be used. Snowberries added to the medium can be added at 0.1% (w/w) to 10% (w/w), but in this example, they were added at 1% (w/w). Next, microorganisms were inoculated into the sterilized medium, and fermentation was performed for 72 hours under conditions of 30 to 40°C with stirring at 150 rpm. The medium composition and types of microorganisms used in Examples 1 to 3 are shown in Table 1 below.

Content (% by weight) ingredient Example 1 Example 2 Example 3 glucose 2 2 2 peptone 0.5 One One yeast extract 0.5 4 0.5 glutamic acid 0.5 - - K ₂ HPO ₄ 0.1 - 0.1 KH ₂ PO ₄ 0.05 - - NaCl - - 0.5 water To 100 To 100 To 100 microbe Bacillus genus
( Bacillus sp. ) Lactobacillus genus
( Lactobacillus sp. ) Leuconostoc genus
( Leuconostoc sp. )

Next, the steps of filtering and obtaining the fermented product were performed. After completion of fermentation, the bacteria and fermentation residue were separated by centrifugation, and snowberry fermentation product was prepared through 0.45㎛ filtration.

Comparative Example 1. Preparation of snowberry extract

Snowberry extract was prepared by hot water extraction of snowberries using a conventional method.

Examples 4 to 6 and Comparative Example 2. Preparation of a composition containing snowberry fermentation broth or extract

A composition containing snowberry fermentation broth or extract was prepared as shown in Table 2 below.

Specifically, Examples 4 to 6 are compositions containing the snowberry fermentation broth of Examples 1 to 3, respectively. Comparative Example 2 is a composition containing the snowberry extract of Comparative Example 1.

ingredient division Content (% by weight) Comparative Example 2 Example 4 Example 5 Example 6 water Distilled water Up to 100 Up to 100 Up to 100 Up to 100 Butylene glycol moisturizer 3 3 3 3 glycerin moisturizer 2 2 2 2 1,2-hexanediol antiseptic 2 2 2 2 Comparative Example 1 extract 10 - - - Example 1 fermented liquid - 10 - - Example 2 fermented liquid - - 10 - Example 3 fermented liquid - - - 10

Experimental Example 1. Confirmation of skin aging prevention efficacy of snowberry fermentation product

In order to confirm the effect of snowberry fermentation on preventing skin aging, MMP-1 expression level analysis was performed according to treatment of the snowberry fermentation prepared above.

Specifically, human dermal fibroblasts were irradiated with ultraviolet rays (UV), treated with the snowberry extract of Comparative Example 1 or the snowberry fermentation product of Examples 1 to 3, and then the expression level of MMP-1 was confirmed. . The treatment concentration of extract or fermented product in each experimental group was 100 ppm. The group that was not treated with ultraviolet rays was used as the untreated control group. In addition, the group treated with only ultraviolet light was used as a negative control group, and the group treated with 10 μM epigallocatechin (EGCG) was used as a positive control group.

Figure 1 shows the results of analyzing the expression level of MMP-1 in human fibroblasts.

As a result, as shown in Figure 1, the negative control group in which human fibroblasts were treated with UV showed a high level of MMP-1 expression. In addition, it was confirmed that when treated with snowberry fermentation product compared to when treated with snowberry extract, the level of MMP-1 expression decreased compared to the negative control group. In particular, snowberry fermentation product 1 of Example 1 using Bacillus microorganisms reduced the expression level of MMP-1 by about 47% compared to the negative control.

From the above results, it can be seen that snowberry fermentation reduces the expression level of MMP-1, showing a significantly excellent anti-aging effect.

Experimental Example 2. Confirmation of the effect of snowberry fermentation on improving skin wrinkles

In order to confirm the effect of the fermented snowberry product on improving skin wrinkles, an analysis of the production of procollagen according to the treatment of the fermented snowberry product prepared above was performed.

Specifically, human fibroblasts were treated with the snowberry extract of Comparative Example 1 or the snowberry fermentation product of Examples 1 to 3, and the relative production rate of procollagen was confirmed. The treatment concentration of the extract or fermented product in each experimental group was 100 ppm. The group not treated with snowberry extract or fermented product was used as a negative control group, and the group treated with TGF-β was used as a positive control group.

Figure 2 shows the results of analyzing the procollagen production rate in human fibroblasts.

As a result, as shown in Figure 2, it was confirmed that when treated with snowberry fermentation product rather than when treated with snowberry extract, the synthesis of procollagen increased compared to the negative control group. In particular, snowberry fermentation product 1 of Example 1 using Bacillus microorganisms had the best effect in increasing procollagen synthesis.

From the above results, it can be seen that fermented snowberry increases procollagen synthesis and exhibits a significantly excellent effect of improving skin wrinkles and improving skin elasticity.

Experimental Example 3. Confirmation of skin whitening effect of fermented snowberry product

In order to confirm the skin whitening effect of the snowberry fermentation product, an experiment was performed to confirm the amount of melanin synthesis according to the treatment of the snowberry fermentation product prepared above.

Specifically, the melanoma B16F10 cell line was treated with the snowberry extract of Comparative Example 1 or the snowberry fermentation product of Examples 1 to 3 with 250 nM of α-MSH, and then the melanin produced in the cells was dissolved to compare melanin content. analyzed. The treatment concentration of extract or fermented product in each experimental group was 100 ppm. The group treated with 50 μg/ml of arbutin was used as a positive control group, and the group treated with only α-MSH was used as a negative control group.

Figure 3 shows the results of analyzing melanin content in melanoma cell lines.

As a result, as shown in Figure 3, when treated with snowberry fermentation product rather than when treated with snowberry extract, the amount of melanin synthesis decreased compared to the negative control group. In particular, snowberry fermentation product 1 of Example 1 using Bacillus microorganisms reduced melanin synthesis the most.

From the above results, it can be confirmed that snowberry fermentation reduces melanin synthesis and exhibits a significantly excellent skin whitening effect.

Experimental Example 4. Confirmation of skin moisturizing effect of snowberry fermentation product

In order to confirm the skin moisturizing effect of the snowberry fermentation product, the expression levels of aquaporin-3 (AQP-3) and filaggrin mRNA according to the treatment of the snowberry fermentation product prepared above were confirmed by performing real-time PCR. did.

Specifically, human keratinocytes were cultured, treated with the snowberry extract of Comparative Example 1 or the snowberry fermentation product of Examples 1 to 3, and mRNA was obtained to compare the expression levels of AQP-3 and filaggrin. analyzed. The treatment concentration of extract or fermented product in each experimental group was 100 ppm. The group that was not treated with snowberry extract or fermented product was used as a negative control group, and the group treated with 1 μM retinoic acid was used as a positive control group. The AQP-3, HAS-3, and Filaggrin primer sequences used in the PCR are listed in Table 3 below.

Primer name Sequence (5'→3') sequence number AQP3_forward AGACAGCCCCTTCAGGATTT One AQP3_reverse TCCCTTGCCCTGAATATCTG 2 Filaggrin_forward AGTGCACTCAGGGGGCTCCACA 3 Filaggrin_reverse CCGGCTTGGCCGTAATGTGT 4

Figure 4 shows the relative expression level of AQP-3 in human keratinocytes.

Figure 5 shows the relative expression level of filaggrin in human keratinocytes.

As a result, as shown in Figures 4 and 5, the expression levels of AQP-3 and filaggrin mRNA increased more in the group treated with snowberry fermentation product than in the group treated with snowberry extract. In particular, snowberry fermentation product 1 of Example 1 using Bacillus microorganisms showed the best increase in AQP-3 and filaggrin expression.

From the above results, it can be seen that the snowberry fermentation product exhibits a significantly excellent skin moisturizing effect and skin barrier strengthening effect.

In summary, it can be confirmed that the cosmetic composition of the present invention contains snowberry fermentation as an active ingredient, thereby exhibiting significantly improved anti-aging, skin wrinkle improvement, skin elasticity enhancement, skin whitening, skin moisturizing, and skin barrier strengthening effects. .

<110> MIGLIM COMPANY LIMITED <120> Composition comprising fermented snowberry and its use for improving skin condition <130> PN143812 <160> 4 <170> PatentIn version 3.2 <210> 1 <211> 20 <212> DNA_RNA <213> Artificial <220> <221> MISC_FEATURE <223> AQP3_forward primer <400> 1 agacagcccc ttcaggattt 20 <210> 2 <211> 20 <212> DNA_RNA <213> Artificial <220> <221> MISC_FEATURE <223> AQP3_reverse primer <400> 2 tcccttgccc tgaatatctg 20 <210> 3 <211> 21 <212> DNA_RNA <213> Artificial <220> <221> MISC_FEATURE <223> Filaggrin_forward primer <400> 3 agtgcactca gggggctcac a 21 <210> 4 <211> 20 <212> DNA_RNA <213> Artificial <220> <221> MISC_FEATURE <223> Filaggrin_reverse primer <400> 4 ccggcttggc cgtaatgtgt 20

Claims (5)

A cosmetic composition for improving skin condition comprising a fermented product obtained by fermenting snowberries with microorganisms,
The microorganism is a microorganism of the genus Lactobacillus ( Lactobacillus sp.) or Leuconostoc sp.,
A cosmetic composition wherein the skin condition improvement includes improving skin wrinkles, improving skin elasticity, moisturizing the skin, strengthening the skin barrier, or a combination of two or more thereof. A cosmetic composition for improving skin condition comprising a fermented product obtained by fermenting snowberries with microorganisms,
The microorganism is a microorganism of the genus Lactobacillus ( Lactobacillus sp.) or Leuconostoc sp.,
A cosmetic composition wherein the skin condition improvement is skin whitening. A food composition for improving skin condition comprising a fermented product obtained by fermenting snowberries with microorganisms,
The microorganism is a microorganism of the genus Lactobacillus ( Lactobacillus sp.) or Leuconostoc sp.,
A food composition wherein the skin condition improvement includes improving skin wrinkles, improving skin elasticity, moisturizing the skin, strengthening the skin barrier, or a combination of two or more thereof. A food composition for improving skin condition comprising a fermented product obtained by fermenting snowberries with microorganisms,
The microorganism is a microorganism of the genus Lactobacillus ( Lactobacillus sp.) or Leuconostoc sp.,
A food composition wherein the improvement in skin condition is skin whitening. delete