KR20240006879A - Composition for anti-aging comprising extracts of Gosorisul lees - Google Patents

Abstract

The present invention relates to an anti-aging composition containing an extract of sake lees from high-sori liquor, which has an effect of suppressing cellular aging caused by oxidative stress, an effect of promoting collagen synthesis, an effect of suppressing elastase activity, and an inflammatory substance. Not only can it effectively prevent and improve skin aging by having a secretion suppressing effect, sebum secretion promoting effect, and whitening effect at the same time, but it can also be useful as a cosmetic because it does not irritate the skin and has excellent safety for the human body.

Description

{Composition for anti-aging comprising extracts of Gosorisul lees}

The present invention relates to an anti-aging composition containing an extract of Gosori sake sake lees. More specifically, the present invention is a sake lees extract that improves skin aging through the effect of suppressing cellular aging caused by oxidative stress, promoting collagen synthesis, suppressing elastase activity, suppressing secretion of inflammatory substances, promoting sebum secretion, and whitening effect. It relates to an anti-aging composition containing as an active ingredient.

Aging causes disharmony in the skin's immune system, which causes various skin diseases. The skin is the organ where aging primarily occurs, and much research has been conducted in the cosmetics field to delay or prevent it. Typically, skin aging begins to progress around the age of 25 and progresses in earnest around the age of 40. Skin aging is divided into extrinsic aging and intrinsic aging. Intrinsic aging is a natural phenomenon caused by degenerative changes that occur in proportion to the progression of time as one ages. Changes in the external appearance of the skin related to skin aging include dry skin, sagging skin, and wrinkles due to decreased moisture and sebum secretion, and skin diseases include seborrheic keratosis and sunspots. As skin ages, the number of fibroblasts in the dermis and their synthetic ability decrease, which results in separation of collagen bundles in the extracellular matrix (ECM), decreased synthesis of point polysaccharide, and collagen and elastic fibers (elastin). ) The dermis shrinks due to a decrease in number and diameter and the breakdown of collagen and elastic fibers, and the epidermis also becomes thinner and drier. The representative trigger of extrinsic aging is reactive oxygen species, which are generated from ultraviolet rays, pollution, inflammation, and the cell's own metabolic process, and act to promote cellular aging. When cellular aging is accelerated, not only does cell regeneration and the ability to synthesize skin components such as collagen, elastin, and hyaluronic acid decrease, but also the responsiveness to external growth factors decreases due to a decrease in growth factor receptors due to aging, thereby activating cells. There are many cases where growth factors do not work properly. In fact, research results continue to report that fibroblasts with advanced cellular senescence do not respond to EGF treatment as their receptors for EGF (epidermal growth factor), a representative skin activation factor, decreases (Hidenori Shiraha et al, JBC, 275 ;19343-19351, 2000). Therefore, it can be inferred that suppressing cellular aging is a major effective skin aging improvement strategy in order to properly respond to cell activating factors such as EGF.

Meanwhile, Gosorisul is a traditional Jeju liquor and is also called Omegisul. “Omegi rice cake” is made by kneading millet alone or with rice, boiling it in water, and going through a fermentation process using malt or yeast. Afterwards, soju is distilled using a gosori, an earthenware vessel for boiling soju, and the remaining dregs become sake lees. In the case of this type of sake lees, it was a discarded resource as a brewing by-product, and little research on its physiological activity has been conducted so far.

Accordingly, the present inventors continued research to develop a natural anti-aging composition with high stability and safety and no skin side effects. As a result, the results showed that the sake lees extract from Gosori Sul has an effect of suppressing cellular aging caused by oxidative stress, promoting collagen synthesis, By discovering that it can simultaneously exhibit an elastase activity inhibition effect, an inflammatory substance secretion inhibition effect, a sebum secretion promotion effect, and a whitening effect, it was discovered that it is a useful ingredient for anti-aging, and the present invention was completed.

One object of the present invention is to provide an anti-aging composition containing sake lees extract from Gosori Sul as an active ingredient.

Another object of the present invention is to provide a cosmetic composition containing the anti-aging composition.

Another object of the present invention is to provide a method for preventing or improving skin aging using the composition.

The present invention relates to an anti-aging composition containing sake lees extract from Gosori Sul as an active ingredient. More specifically, the present invention relates to a cosmetic composition containing sake lees extract from Gosori Sul as an active ingredient. The composition is free from skin irritation and cytotoxicity and has excellent human safety, while suppressing cellular aging caused by oxidative stress. , It can improve skin aging by promoting collagen synthesis, suppressing elastase activity, suppressing secretion of inflammatory substances, promoting sebum secretion, and effectively suppressing melanin production.

In one aspect, the present invention provides an anti-aging composition containing an extract of sake lees from Gosori Sul as an active ingredient.

Gosori alcohol of the present invention is a traditional liquor of Jeju Island, mixed with millet and rice, kneaded, boiled in boiling water, fermented using malt and yeast, and then distilled in Gosori, an earthenware vessel for boiling soju.

The Gosori liquor, unlike traditional liquor in other regions, is characterized by mixing millet and rice, kneading and fermenting. The mixture of millet and rice is at a weight ratio of 1:9 to 9:1, preferably 2:8 to 8:2, more preferably 3:7 to 7:3, even more preferably 5. It can be achieved with a weight ratio of :5.

The millet is the grain remaining after removing the fruit of the millet. It is rich in protein, second only to soybeans among grains, contains more unsaturated fatty acids than rice or barley, contains carotene that is not found in ordinary grains, and has a savory taste. The rice is the remaining grains after removing the fruit of the rice plant, and unlike the millet, it contains a lot of starch.

At this time, if the content of the rice is less than the above range, fermentation by malt and yeast is not sufficient, making it difficult to manufacture high-sodium alcohol, and almost no useful substances are present in the sake lees of high-sori alcohol, resulting in the effects described below. There is a problem with it becoming weak. On the other hand, when the content of rice exceeds the above range, there is a problem that the sensory properties of the high-sodium alcohol produced are reduced due to excessive fermentation. However, since there are sufficient useful substances in the sake lees of the high-sori liquor, the effects described below are obtained. It has the advantage of being sufficiently expressed.

In addition, if the content of the millet is less than the above range, the sensory properties (particularly the savory taste) due to the content of the millet are reduced, which not only poses a problem in the sensory quality of the high-soured liquor produced, but also the useful content contained in the millet reduces the high-sourized liquor. There is a problem in that very little remains in the sake lees, making the effects described below very weak. On the other hand, when the content of millet exceeds the above range, there is an advantage that the sensory quality of gosori liquor produced is improved due to the increase in savory taste due to the inclusion of millet, and the above-mentioned useful components contained in millet are used as sake lees of gosori liquor. There is an advantage that the effect described below is excellent because a large amount remains in the rice, but there is a problem that the quality of Gosori liquor is reduced because the rice content is relatively reduced and fermentation does not occur sufficiently.

Therefore, it is important to produce gosori alcohol with the content of millet and rice described above, and furthermore, the purpose of the present invention can be achieved by using the sake lees of gosori liquor.

In the production of the high-solid liquor, the yeast is preferably a fermenting microorganism of the genus Aspergillus ( Aspergillus spp.), and the yeast is preferably a fermenting microorganism of the genus Saccharomyces ( Sacchraromycess spp.).

When producing the above-mentioned high-solid liquor, fermentation using nuruk and yeast is preferably carried out at 20°C to 30°C for 7 to 10 days. The quality of Gosori liquor is maintained due to sufficient fermentation under the above-mentioned temperature and time conditions.

The sake lees of the high-sori liquor of the present invention is the liquor residue remaining after producing the high-sori liquor.

In the present invention, the “sake lees extract of Gosori liquor” is extracted from the sake lees of Gosori liquor, and the “extract” is obtained by immersing the target substance in various solvents and then extracting it at room temperature or at a heated state for a certain period of time. It refers to the results such as the liquid component obtained by fractionation and the solid component obtained by removing the solvent from the liquid component. In addition, in addition to the above results, it can be comprehensively interpreted to include all dilutions of the results, concentrates thereof, crude preparations, purifications, etc. of the above results.

The sake lees extract of the highly fermented liquor of the present invention can be extracted using a conventional solvent known in the art, and the extracted liquid can be used in liquid form or after being concentrated and/or dried. At this time, the solvent used is water, anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, propanol, butanol, etc.), a mixed solvent of the lower alcohol and water, acetone, ethyl acetate, chloroform, or 1,3- Any solvent of butylene glycol can be used. Preferably, extraction may be performed using methanol, ethanol, or butanol. However, it is not particularly limited to this, and the degree of extraction and loss of the active ingredients of the extract may vary depending on the organic solvent used for extraction, so it is desirable to select and use an appropriate organic solvent.

The extraction method is not particularly limited to this, but includes cold needle extraction, ultrasonic extraction, or reflux cooling extraction, and is preferably performed using the ultrasonic extraction method.

In addition, the sake lees extract from the high-steaming liquor can be obtained through a conventional purification process in addition to extraction using the extraction solvent. For example, through various additional purification methods, such as separation using ultrafiltration membranes with a certain molecular weight cut-off value, separation by various chromatographs (designed for separation according to size, charge, hydrophobicity, or affinity). Sake lees extract can also be obtained through the obtained fraction.

As a specific example, the sake lees of Gosori sake are shaded and ground, then 0.5 to 20 times the weight of the dried sample, preferably 1 to 15 times the amount of water, C1 to C4 lower alcohol such as ethanol, methanol, etc. Or, using a mixed solvent of water and ethanol at room temperature for about 20 to 70 hours, preferably 40 to 50 hours, using extraction methods such as stirred extraction, hot water extraction, cold immersion extraction, reflux cooling extraction, or ultrasonic extraction. Alternatively, the extract obtained after ultrasonic extraction can be filtered, concentrated under reduced pressure, or dried to obtain a sake lees extract.

In the present invention, the “anti-aging” refers to reducing or preventing aging or death of skin cells or damage caused by external pressure, fine or coarse wrinkles, decreased skin elasticity, etc., through cell aging inhibition, prevention, and regenerative effects. Or it means improving. Therefore, “anti-aging” in the present invention includes preventing or improving skin wrinkles, preventing or improving skin elasticity, promoting skin regeneration, preventing, reducing, or improving skin inflammation, and preventing or improving skin whitening. can be considered. Additionally, the term “anti-aging” may be used interchangeably with aging improvement or skin aging improvement in this specification.

In one specific embodiment, the Gosori liquor sake lees extract of the present invention has an effect of suppressing cellular aging caused by oxidative stress, an effect of promoting collagen synthesis, an effect of suppressing elastase activity, an effect of suppressing secretion of inflammatory substances, an effect of promoting sebum secretion, and a whitening effect. It is very useful in preventing, improving and treating skin aging.

In the present invention, the term “prevention” refers to suppressing or minimizing cell aging caused by oxidative stress, suppressing or minimizing skin wrinkles by promoting the synthesis of skin collagen, suppressing the activity of elastase, and promoting sebum secretion. , increases skin elasticity by promoting the synthesis of skin collagen, suppressing the activity of elastase, and promoting sebum secretion, increases anti-inflammation of the skin by suppressing or reducing the secretion of inflammatory substances, or increases the production of melanin. It refers to suppressing and minimizing skin aging by increasing skin whitening.

In the present invention, the term “improvement” or “treatment” refers to suppressing or minimizing cell aging caused by oxidative stress, or reducing skin wrinkles by promoting the synthesis of skin collagen, suppressing the activity of elastase, and promoting sebum secretion. Inhibit or minimize, increase skin elasticity by promoting the synthesis of skin collagen, suppressing the activity of elastase, and promoting sebum secretion, or increasing anti-inflammation of the skin by suppressing or reducing the secretion of inflammatory substances, It refers to preventing skin aging and returning the skin condition to its previous state by increasing the production of melanin and increasing skin whitening.

In the present invention, the cells include all cells existing in vivo or cultured in vitro . The cells can be classified in various ways based on cell shape and location, and examples include skin cells (epidermis, dermis, dermal papilla cells, melanin, etc.), fibroblasts, adipocytes, stem cells, and vascular cells. .

In one specific example, cellular senescence was induced in human fibroblasts and the degree of cellular senescence was examined depending on the concentration of the high-solid sake lees extract of the present invention. As a result, as the concentration of the highly-sulliary sake lees extract increased, cellular senescence decreased. It was confirmed that the sake lees extract from Gosori Sul has an anti-aging effect (see Figure 1).

In another specific example, as a result of examining the rate of increase in collagen synthesis according to the concentration of the sake lees extract of high-sori liquor using human fibroblasts, it was found that collagen synthesis was promoted as the concentration of the sake lees extract of high-sori liquor increased. It was confirmed that sake lees extract exhibits wrinkle improvement and skin regeneration effects by synthesizing collagen (see Figure 2).

In another specific example, as a result of examining the degree of inhibition of elastase activity according to the concentration of the sake lees extract of high-sori liquor using an elastase analysis kit, it was found that elastase activity was inhibited by the sake lees extract of high-sori liquor. It was shown to decrease, and it was confirmed that the sake lees extract of Gosori liquor exhibits a wrinkle-improving effect by inhibiting elastase activity (see Figure 3).

In another specific example, an inflammatory response was induced in mouse macrophages and the degree of secretion of inflammatory substances (nitrite) according to the concentration of the sake lees extract of high-sori liquor was examined. As a result, nitrite (nitrite) was secreted by the sake lees extract of high-sori liquor. ) production was found to be reduced, and it was confirmed that the sake lees extract from Gosori Sul has anti-inflammatory and anti-aging effects by suppressing the secretion of inflammatory substances (see Figure 4).

In another specific example, as a result of examining the degree of sebum secretion according to the concentration of sake lees extract of high-sori liquor using human sebaceous cells, it was found that sebum secretion was promoted as the concentration of sake lees extract of high-sori liquor increased. , it was confirmed that the sake lees extract from Gosori Sul promotes sebum secretion, thereby improving wrinkles and suppressing aging (see Figure 5).

In another specific example, as a result of examining the degree of melanin production according to the concentration of the sake lees extract of Gosori liquor using human melanocytes, it was found that melanin production was suppressed by the sake lees extract of Gosori liquor. It was confirmed that sake lees extract exhibits a whitening effect by suppressing melanin production (see Figure 6).

The anti-aging composition containing the sake lees extract of Gosori sake of the present invention as an active ingredient contains 0.0001 to 20.0% by weight, preferably 0.001 to 10.0% by weight, of the sake lees extract of Gosori sake based on the total weight of the composition. . If the content of the sake lees extract of the Gosori liquor is less than 0.0001% by weight, anti-aging effect cannot be expected, and if it exceeds 20.0% by weight, the increase in anti-aging effect with increasing content is minimal and difficulties arise in manufacturing the formulation. .

In one specific embodiment, the anti-aging composition of the present invention may be provided as an anti-aging cosmetic composition containing an extract of sake lees from Gosori Sul as an active ingredient.

The cosmetic composition may preferably be in the form of a functional cosmetic, and the functionality may be skin anti-aging, including skin wrinkle improvement, skin antioxidant, skin anti-inflammatory, skin elasticity improvement, skin regeneration, skin whitening, etc.

The sake lees extract of Gosori Sul is as described above, and the sake lees extract of Gosori Sul has skin anti-aging effects such as skin wrinkle improvement effect, skin antioxidant effect, skin elasticity enhancement effect, skin anti-inflammatory effect, and skin whitening effect. Because it has an effect, it can be usefully used as a cosmetic composition for skin anti-aging purposes such as improving skin wrinkles, skin antioxidant, improving skin elasticity, skin anti-inflammatory, skin regeneration, and skin whitening.

The cosmetic composition may additionally contain ingredients commonly used in cosmetic compositions in addition to the sake lees extract of Gosori wine as an active ingredient with anti-aging effect. For example, it may include conventional auxiliaries and carriers such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavorings.

The cosmetic composition can be manufactured in any formulation commonly manufactured in the art. For example, it can be formulated as a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation or spray. More specifically, it can be manufactured in formulations such as nutritional cream, astringent lotion, softening lotion, lotion, essence, nutritional gel, or massage cream. However, it is not particularly limited thereto.

When the formulation of the cosmetic composition is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide are used as carrier ingredients. It can be used.

When the formulation of the cosmetic composition is powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder can be used as the carrier ingredient. In particular, in the case of spray, chlorofluorohydrocarbon, May contain propellants such as propane/butane or dimethyl ether.

When the formulation of the cosmetic composition is a solution or emulsion, a solvent, solubilizing agent, or emulsifying agent may be used as a carrier component. For example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic esters, polyethylene glycol or fatty acid esters of sorbitan can be used. there is.

When the formulation of the cosmetic composition is a suspension, the carrier ingredients include water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester, and miso. Crystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracant may be used.

When the formulation of the cosmetic composition is a cleansing agent containing a surfactant, the carrier ingredients include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, and fatty acid. Amide ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester can be used.

The cosmetic composition containing the sake lees extract of Gosori sake of the present invention as an active ingredient contains 0.0001 to 20.0% by weight, preferably 0.001 to 10.0% by weight, of the sake lees extract of Gosori alcohol based on the total weight of the composition. If the content of the sake lees extract of the Gosori liquor is less than 0.0001% by weight, anti-aging effect cannot be expected, and if it exceeds 20.0% by weight, the increase in the effect due to increase in content is insignificant and difficulties arise in manufacturing cosmetic formulations. .

Meanwhile, the sake lees extract from Gosori liquor according to the present invention is a natural substance that is harmless to the human body and has almost no toxicity or side effects, so it can be safely used even for long-term use, and can be safely applied to cosmetic compositions containing it.

The sake lees extract, which is an active ingredient of the present invention, does not irritate the skin and has excellent human safety, but also has an effect of inhibiting cell aging caused by oxidative stress, promoting collagen synthesis, inhibiting elastase activity, inhibiting secretion of inflammatory substances, promoting sebum secretion, and It can be usefully used as an anti-aging cosmetic composition that effectively improves skin aging by simultaneously having a whitening effect.

Figure 1 is a diagram showing the cell senescence inhibitory effect of sake lees extract when oxidative stress is induced in normal human fibroblasts according to an embodiment of the present invention.
Figure 2 is a diagram showing the collagen synthesis promoting effect of sake lees extract in normal human fibroblasts according to an embodiment of the present invention.
Figure 3 is a diagram showing the elastase activity inhibitory effect of sake lees extract according to an embodiment of the present invention.
Figure 4 is a diagram showing the inhibitory effect of the sake lees extract on the secretion of inflammatory substances when an inflammatory response is induced in mouse macrophages according to an embodiment of the present invention.
Figure 5 is a diagram showing the effect of promoting sebum secretion after treating human sebum cells with sake lees extract according to an embodiment of the present invention.
Figure 6 is a diagram showing the whitening effect of sake lees extract on human melanocytes according to an embodiment of the present invention.

Hereinafter, to aid understanding of the present invention, it will be described in detail through examples. However, the embodiments according to the present invention may be modified into various other forms, and the scope of the present invention should not be construed as being limited to the following embodiments. Embodiments of the present invention are provided to more completely explain the present invention to those skilled in the art.

Example 1: Preparation of Gosori Sake Sake Lid Extract

After mixing millet and rice in a weight ratio of 5:5, kneading it in hot purified water and boiling it in boiling water, add malt ( Aspergillus spp.) and yeast ( Saccharomyces spp.) and knead at 20℃ to 30℃ for 8 days. After fermentation, it was distilled using Gosori, an earthenware vessel used to boil soju, to produce Gosori liquor, a traditional liquor of Jeju Island. After producing the high-sori liquor, the remaining liquor residue (sake lees) was recovered, dried in the shade at 20°C to 35°C, and then ground to a particle size of 1 mm or less. Afterwards, 100 g of pulverized sake lees powder was immersed in 70% ethanol solvent and subjected to ultrasonic extraction for 48 hours, and the obtained extract was filtered using filter paper (Advantec, No. 2). The filtrate was concentrated under reduced pressure to prepare a sake lees extract.

Example 2: Confirmation of the inhibitory effect of cellular senescence induced by oxidative stress in human fibroblasts

In order to investigate the inhibitory effect of the sake lees extract on cellular aging induced by oxidative stress, the following experiment was conducted. Human dermal fibroblasts (HDFs) were inoculated at 1 × 10 ⁵ per well in a 6-well plate containing DMEM medium supplemented with 10% FBS (fetal bovine serum), and incubated in a 5% concentration CO ₂ incubator. Cultured for 24 hours at 37°C. Afterwards, the test substance and hydrogen peroxide (H ₂ O ₂ 300 μM), which can induce oxidative stress, were dissolved together in DMEM medium without FBS and then treated. EGCG (Epigallocatechin-3-gallate) was used as a positive control to inhibit cellular senescence. After 72 hours, the cell culture supernatant was removed, the cells were detached by treatment with trypsin-EDTA, and the cells were fluorescently stained with Dojindo's SPiDER-ßGal for 15 minutes and flow cytometry (FACS Calibur, Becton Dickinson, San Francisco). Jose, CA), and cells were counted. Changes in cellular aging were observed by confirming the proportion of cells showing aging-related β-galactosidase activity. The results are shown in Figure 1.

As shown in Figure 1, it was confirmed that sake lees extract reduced the increased SA-β-gal activity caused by hydrogen peroxide treatment in human fibroblasts. From this, it was found that sake lees extract had an excellent effect in suppressing cellular aging caused by oxidative stress in human fibroblasts in a concentration-dependent manner.

Example 3: Confirmation of collagen synthesis promoting effect in human fibroblasts

To investigate the effect of sake lees extract on promoting collagen synthesis, the following experiment was conducted. Human fibroblasts were seeded at 2 × 10 ⁴ per well in a 24-well plate containing DMEM medium supplemented with 10% FBS, and then cultured in a 5% CO ₂ incubator at 37°C for 24 hours. Afterwards, the test substance was dissolved in DMEM medium without FBS and then treated. Transforming growth factor-β (TGF-β) was used as a positive control for promoting collagen synthesis. After 72 hours, cell culture supernatants were collected to prepare samples. The amount of collagen synthesis in the sample prepared above was measured by measuring procollagen type I C-peptide (PICP, Type I C-peptide) using TAKARA's collagen measurement kit (MK101, Procollagen Type I C-peptide EIA kit). The amount was measured. The results are shown in Figure 2.

As shown in Figure 2, it was confirmed that the sake lees extract promoted collagen synthesis in the human fibroblast culture supernatant.

Example 4: Confirmation of elastase activity inhibition effect

In order to investigate the inhibitory effect of sake lees extract on elastase activity, the following experiment was conducted. P.C was used as a positive control as an elastase activity inhibitor, and the level of elastase activity of the test substance was measured using a 96-well black plate and Invitrogen's elastase activity measurement kit (EnzChek™ Elastase Assay Kit). did. The results are shown in Figure 3.

As shown in Figure 3, it was confirmed that the sake lees extract inhibited elastase activity.

Example 5: Confirmation of inhibitory effect on secretion of inflammation-inducing substances in mouse macrophages

In order to investigate the inhibitory effect of sake lees extract on the secretion of nitric oxide, an inflammation-inducing substance, the following experiment was conducted. Mouse macrophages (RAW264.7, macrophage) were inoculated at 2 × 10 ⁵ per well in a 24-well plate containing RPMI medium supplemented with 10% FBS (fetal bovine serum), and then incubated in a 5% concentration CO ₂ incubator. Cultured for 24 hours at 37°C. Afterwards, the test substance was dissolved in RPMI medium supplemented with 2% FBS, pretreated for 2 hours, and then LPS (Lipopolysaccharide 100 ng/mL), which can induce an inflammatory response, was treated in each well. 1400W was used as a positive control for inhibition of nitric oxide secretion. After 24 hours, cell culture supernatants were collected to prepare samples. The sample prepared above was reacted with Griess reagent to quantify NO ^{2 -} (nitrite), a stabilized oxidizing substance of nitric oxide, thereby quantifying the produced nitric oxide. The results are shown in Figure 4.

As shown in Figure 4, it was confirmed that sake lees extract reduced the increased nitrite secretion due to LPS treatment in mouse macrophages. From this, it was found that sake lees extract was effective in suppressing the secretion of inflammatory substances caused by inducing an inflammatory response in mouse macrophages.

Example 6: Confirmation of sebum secretion promoting effect in human sebum cells

In order to determine the effect of sake lees extract on promoting sebum secretion, the following experiment was conducted. Human sebocytes were inoculated at 1 × 10 ⁴ per well in a 24-well plate containing sebum cell culture medium, and then cultured in a 5% CO ₂ incubator at 37°C for 24 hours. Afterwards, the test substance was dissolved in the culture medium and treated. After 48 hours, the cell culture supernatant was removed and fixed with a 4% (v/v) paraformaldehyde solution. Palmitic acid was used as a positive control for promoting sebum secretion. To confirm the sebum synthesized in sebum cells, the cells were stained with Nile red for 15 minutes at room temperature, and then the nuclei were stained with Hoechst 33342 to confirm the total number of cells, and the degree of sebum secretion was observed under a fluorescence microscope. Micrographs were analyzed with ImageJ, and the results are shown in Figure 5.

As shown in Figure 5, it was confirmed that the sake lees extract promoted sebum secretion from human sebum cells.

Example 7: Confirmation of whitening effect in human melanocytes

In order to determine the whitening effect of sake lees extract, the following experiment was conducted. Human melanocytes were inoculated at 1.5 × 10 ⁵ per well in a 6-well plate containing melanocyte culture medium, and then cultured in a 5% CO ₂ incubator at 37°C for 24 hours. Afterwards, the test substance was dissolved in the culture medium and treated. After 5 days, the cell culture supernatant was removed and the cells were collected with a Cell Scraper. Kojic acid was used as a positive control for inhibiting melanin production. 1M sodium hydroxide solution was added to the collected cells, and the absorbance of the intracellular melanin solution was measured at 450 nm. The results are shown in Figure 6.

As shown in Figure 6, it was confirmed that the sake lees extract exhibited a whitening effect by suppressing melanin production in human melanocytes.

Example 6: Experiment to confirm the safety of sake lees extract on human skin

In order to confirm whether the sake lees extract, which was found to have excellent anti-aging effect according to the present invention, is safe for human skin, an external skin preparation containing the sake lees extract was prepared and a skin safety verification experiment was performed.

An external skin preparation containing 1% of the sake lees extract of Example 1 was prepared with the ingredient contents shown in Table 1 below. To prepare an external skin agent, purified water, glycerin, and butylene glycol were mixed and dissolved at 70°C (aqueous part), and the remaining ingredients except the above three ingredients and trimethanolamine were dissolved at 70°C (oil part). The oil part was added to the water part and stirred with a homomixer (Tokushu Kika, Japan) for primary emulsification, and trimethanolamine was finally added thereto. After removing the bubbles generated in the mixed solution, it was cooled to room temperature to prepare an external skin preparation.

ingredient content (%) control group test group Purified water 72 71 glycerin 8.0 8.0 Butylene glycol 4.0 4.0 Gosori Sake Sake Lid Extract 0 1.0 Caprylic Capri Triglyceride 8.0 8.0 squalane 5.0 5.0 Cetearyl Glucuside 1.5 1.5 Sorbitan Stearate 0.4 0.4 cetearyl alcohol 1.0 1.0 Trimethanolamine 0.1 0.1 total weight 100 100

We measured whether the sake lees extract irritates the skin by applying an occlusive patch to the back of 30 healthy adults for 24 hours using each skin external preparation prepared as above.

The patch method used a chamber (IQ Ultimate™) from Chemotechnique MB Diagnostics AB. 20 μL of each of the above skin external preparations was taken into the chamber and an occlusive patch was applied for 24 hours. Skin reaction was observed 30 minutes and 24 hours after removal of the patch, and the presence and degree of skin irritation was visually evaluated according to the evaluation criteria. Based on Frosch & Kligman (1979) and CTFA guideline (2007), skin After calculating the irritation evaluation index, the average skin irritation index of each external skin preparation was calculated using the following empirical formula 1, and the skin irritation was evaluated based on the skin irritation index in Table 3 classified by applying the Draize method.

[Empirical Formula 1]

Average skin irritation index calculation formula

∑ [(Grade × Number of subjects who responded)/ (4; Highest grade × Total number of subjects)]÷ Number of evaluations

sign Grade Irritation evaluation - 0 no response + One Mild but clearly visible erythema reaction ++ 2 Erythematous reaction with clear erythema and papules or edema +++ 3 Strong erythematous reaction with edema and papules ++++ 4 Strong erythematous reaction with edema and blisters

skin irritation index Determination of skin irritation level 0.00-0.25 Non-irritating 0.26-1.00 mild irritant 1.01-2.50 moderate irritant 2.51-4.00 strong irritant

test
matter Number of subjects who responded average
Skin irritation index 30 minutes later 24 hours later One 2 3 4 One 2 3 4 control group - - - - - - - - 0.00 test group - - - - - - - - 0.00 Number of subjects 30 30 30 30 30 30 30 30

In Table 4, no stimulation response occurred in both the control and test groups. When calculated according to the formula described above, the average reactivity was 0.00, so the skin external preparation containing sake lees extract (test group) was determined to be non-irritating and safe for human skin, like the control skin external preparation.

Preparation Example 1: Cosmetic formulation

Preparation Example 1-1: Soft lotion

As shown in Table 5 below, a softening lotion containing Gosori Sake lees extract as an active ingredient was prepared according to a conventional method.

ingredient weight% Gosori Sake Sake Lid Extract 0.01 glycerin 3.0 butylene glycol 2.0 propylene glycol 2.0 Carboxy vinyl polymer 0.1 ethanol 10.0 Triethanolamine 0.1 Preservatives, trace colors, trace fragrances and trace amounts of purified water 82.79 sum 100.0

Preparation Example 1-2: Nutritional lotion

As shown in Table 6 below, a nutritious lotion containing Gosori liquor sake lees extract as an active ingredient was prepared according to a conventional method.

ingredient weight% Gosori Sake Sake Lid Extract 0.01 beeswax 4.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.5 liquid paraffin 5.0 squalane 5.0 Caprylic/Capric Triglyceride 5.0 glycerin 3.0 butylene glycol 3.0 propylene glycol 3.0 Carboxy vinyl polymer 0.1 Triethanolamine 0.2 Preservatives, trace pigments, trace fragrances and trace purified water 69.69 sum 100.0

Manufacturing Example 1-3: Nutritional cream

As shown in Table 7 below, a nutritional cream containing Gosori Sake lees extract as an active ingredient was prepared according to a conventional method.

ingredient weight% Gosori Sake Sake Lid Extract 0.01 beeswax 10.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.5 liquid paraffin 10.0 squalane 5.0 Caprylic/Capric Triglyceride 5.0 glycerin 5.0 butylene glycol 3.0 propylene glycol 3.0 Triethanolamine 0.2 Preservatives, trace pigments, trace fragrances and trace purified water 56.79 sum 100.0

Manufacturing Example 1-4: Massage Cream

As shown in Table 8 below, a nutritional cream containing Gosori Sake lees extract as an active ingredient was prepared according to a conventional method.

ingredient weight% Gosori Sake Sake Lid Extract 0.01 beeswax 10.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.8 liquid paraffin 40.0 squalane 5.0 Caprylic/Capric Triglyceride 4.0 glycerin 5.0 butylene glycol 3.0 propylene glycol 3.0 Triethanolamine 0.2 Preservatives, trace pigments, trace fragrances and trace purified water 27.49 sum 100.0

Manufacturing Example 1-5: Pack

As shown in Table 9 below, a pack containing Gosori Sake lees extract as an active ingredient was prepared according to a conventional method.

ingredient weight% Gosori Sake Sake Lid Extract 0.01 polyvinyl alcohol 13.0 Sodium Carboxymethyl Cellulose 0.2 allantoin 0.1 ethanol 5.0 Nonylphenyl ether 0.3 Preservatives, trace pigments, trace fragrances and trace purified water 81.39 sum 100.0

Claims (5)

An anti-aging cosmetic composition containing sake lees extract from Gosori liquor as an active ingredient. The anti-aging cosmetic according to claim 1, wherein the sake lees of the high-sori alcohol is the liquor residue remaining after mixing millet and rice, kneading them, boiling them in boiling water, fermenting them with malt and yeast, and then distilling them with high-sori alcohol. Composition. The method of claim 1, wherein the anti-aging is at least one selected from the group consisting of preventing or improving skin wrinkles, preventing or improving skin elasticity, promoting skin regeneration, preventing or improving skin inflammation, and preventing or improving skin whitening. Characterized by an anti-aging cosmetic composition. The cosmetic composition for preventing or improving aging according to claim 1, wherein the extract is contained in an amount of 0.0001 to 20% by weight based on the total weight of the entire cosmetic composition. The cosmetic composition according to claim 1, wherein the cosmetic composition consists of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray. An anti-aging cosmetic composition, characterized in that it is any one of the group.