KR102238329B1 - Skin funtional composition containing fermented natural product complex by lactic acid - Google Patents

Abstract

The present invention relates to a composition that enables the improvement and enhancement of skin functionality, including a fermented product obtained by fermenting a mixture of a plurality of natural substances with specific lactic acid bacteria, and in one aspect, Lactobacillus sa K ( Lactobacillus sakei ) As a cosmetic composition, pharmaceutical composition or health food composition containing fermented product as an active ingredient, Lactobacillus sakei is Lactobacillus Sake ATCC 15521, Lactobacillus Sake MGB0632MGB0522 Tobacillus Sake It relates to a composition that is at least one selected from the group consisting of MGB0522MGB0522.

Description

Skin functional composition containing a natural mixture of fermented lactic acid bacteria {SKIN FUNTIONAL COMPOSITION CONTAINING FERMENTED NATURAL PRODUCT COMPLEX BY LACTIC ACID}

The present invention relates to a composition that enables improvement and enhancement of skin functionality, including a fermented product obtained by fermenting a mixture of a plurality of natural products with specific lactic acid bacteria.

The epidermal layer contains moisture in about 70% in the base layer and the play layer, 30-50% in the granule layer, and 15-30% in the stratum corneum. That is, the moisture content in the epidermis exhibits a skin water gradient that gradually decreases toward the top, and through this moisture gradient, the skin is subjected to continuous osmotic stress. In addition, since the skin exists in the outermost layer of the human body and is chronically exposed to air pollution, dry environment, ultraviolet rays, wind, chemicals, etc., it dries easily and reduces gloss and elasticity, which in turn promotes skin aging.

On the other hand, free radicals, which are known as a major cause of skin aging, cause tissue damage to skin cells, resulting in oxidative stress. The resulting oxidative stress damages biological components, such as destruction of lipid components between cells and promoting melanin production, and appears as a result of decreased skin elasticity and the expression of spots and freckles.

Antioxidants that can control free radicals include butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT), but most of them are regulated due to stability controversy. Accordingly, the necessity of developing new natural antioxidants is increasing according to the recent trend of nature-oriented and environment-friendly consumption.

Republic of Korea Patent Publication No. 10-2006-0106310

It is an object of the present invention to provide a composition comprising a fermented product of natural substances having an excellent moisturizing effect, improving and enhancing skin elasticity, and improving wrinkles.

In order to achieve the above object, the present invention is a cosmetic composition for moisturizing, a pharmaceutical composition comprising a fermented product of Lactobacillus sakei of a mixture of horseback riding, dermis, gardenia, oleracea and licorice as an active ingredient in one aspect. And a health food composition, wherein the Lactobacillus sakei (Lactobacillus sakei) is one or more compositions selected from the group consisting of Lactobacillus Sake ATCC 15521, Lactobacillus Sake MGB0632 and Lactobacillus Sake MGB0522.

In another aspect, the present invention comprises a fermented product of Lactobacillus sakei of a mixture of horseback riding, dermis, gardenia, oleracea and licorice as an active ingredient, a cosmetic composition for improving wrinkles and preventing wrinkles, a pharmaceutical composition, and a health food Regarding the composition, the Lactobacillus sakei (Lactobacillus sakei) provides a composition of one or more selected from the group consisting of Lactobacillus Sake ATCC 15521, Lactobacillus Sake MGB0632 and Lactobacillus Sake MGB0522.

In another aspect, the present invention is a cosmetic composition for improving skin elasticity and preventing reduction of skin elasticity, comprising a fermented product of Lactobacillus sakei of a mixture of horse riding, dermis, gardenia, oleracea and licorice as an active ingredient, Regarding the composition and health food composition, the Lactobacillus sakei (Lactobacillus sakei) provides a composition of one or more selected from the group consisting of Lactobacillus Sake ATCC 15521, Lactobacillus Sake MGB0632 and Lactobacillus Sake MGB0522.

The composition as an aspect of the present invention has excellent antioxidant activity, including fermented products of natural mixtures of lactic acid bacteria as an active ingredient. Specifically, the active ingredients contained in the composition of one aspect of the present invention can inhibit elastase and collagenase, which are the causes of skin wrinkle formation and elasticity reduction, and increase the synthesis of procollagen, a precursor of collagen, Since it has the advantage of being able to perform the effect of improving skin wrinkles and increasing elasticity more actively, it can be usefully used as a cosmetic composition or health food composition for improving skin wrinkles and improving elasticity, and for preventing skin wrinkles. It can also be used as a pharmaceutical composition. In addition, the active ingredient contained in the composition, which is one aspect of the present invention, has a moisturizing effect that can increase skin moisture content and improve skin texture, so it can be usefully used as a cosmetic composition or health food composition for improving skin moisturization. It can also be used as a pharmaceutical composition for preventing or treating diseases such as psoriasis and atopy. In addition to these advantages, the composition, which is one aspect of the present invention, contains a natural-derived ingredient as an active ingredient, and thus has no side effects and does not cause cytotoxicity even when used for a long period of time.

In addition, when the composition of one aspect of the present invention contains a fermented product fermented with one or more strains selected from the group consisting of Lactobacillus SAK ATCC 15521, Lactobacillus SAK MGB0632, and Lactobacillus SAK MGB0522, it is superior to the prior art. It has moisturizing activity and anti-aging activity (wrinkle improvement, elasticity enhancement, etc.), and especially, in the case of fermented products containing all of the above three strains, it shows remarkably excellent moisturizing and anti-aging effects.

1 and 2 are results confirming the antioxidant effect of the fermented product, which is an active ingredient of the composition of the present invention (FIG. 1: DPPH assay, FIG. 2: ABTS assay).
3 to 5 are results of evaluating the cytotoxicity of fermented products, which are active ingredients of the composition of the present invention.
6 is a view confirming the degree to which the fermented product, which is an active ingredient of the composition of the present invention, inhibits the activity of elastase.
7 is a confirmation that the fermented product, which is an active ingredient of the composition of the present invention, inhibits the activity of collagenase.
Figure 8 is a check of the procollagen synthesis ability of the fermented product, which is an active ingredient of the composition of the present invention.
9 is a result of confirming that the production amount of hyaluronic acid increases according to the fermented product, which is an active ingredient of the composition of the present invention.
10 is a result of clinical evaluation of cosmetics containing fermented products, which are active ingredients of the composition of the present invention.

In the present specification, "equestrian (升麻, Cimicifugae Rhizoma )" may refer to Cimicifugaheracleifolia belonging to the Ranunculus family or the root of a plant in the same genus.

In the present specification, "dermis (陳皮)" may refer to a mandarin orange (Citrus unshiu Markovich, Citrus reticulata Blancoe) or a mature pericarp of a plant of the same genus.

In the present specification, "Liquorice (甘草)" may be a perennial herbaceous plant having the scientific name Glycyrrhiza uralensis.

In the present specification, "Gardenia (梔子)" has the same scientific name as Gardenia jasminoides and may be an evergreen broad-leaved shrub belonging to the Rubiaceae family.

In the present specification, "Obaeja (五倍子)" may be a worm house formed by parasitic on the leaves of Rhus javanica Linne of the Rhus javanica Linne family of the five-baeja worms of the five-baeja aphid family.

In the present specification, the "mixture" includes all of the horseback riding, dermis, licorice, gardenia, and oleracea, and specifically, may be a hot water extract of horseback riding, dermis, licorice, gardenia, and oleracea, but is not limited thereto. The above mixture is meant to include both horseback riding, dermis, licorice, gardenia, and extracts obtained after mixing or mixing them after preparing the extracts of each of the five natural products. More specifically, the mixture may contain a relative weight ratio of 2 to 4 horseback riding, 1.5 to 3.5 dermis, 1 to 3 licorice, and 0.5 to 2.5 gardenia with respect to the weight of the five baeja. In the weight ratio, the skin moisturizing and anti-aging effect, which is one object of the present invention, specifically, wrinkle improvement and prevention, skin elasticity improvement and elasticity reduction prevention effect can be maximized. From this point of view, the mixture may contain 1.5 to 3.5 horseback riding, 2 to 3 dermis, 1.5 to 2.5 licorice, and 1 to 2 gardenia relative to the weight of the five baeja. More specifically, the mixture of the present specification may be obtained by extracting the natural product of the above ratio with hot water and then aging it at room temperature for 10 to 12 hours.

In the present specification, “fermentation” refers to a process of decomposing organic matter by putting the lactic acid bacteria and the mixture in the minimum medium required for the propagation of lactic acid bacteria, and the lactic acid bacteria usable in the fermentation process may be Lactobacillus sakei, The lactic acid bacteria minimum medium may be used without limitation, provided that it is a composition known to be capable of cultivating lactic acid bacteria well. Specifically, Lactobacillus sakei of the present specification may be one or more strains selected from the group consisting of Lactobacillus Sake ATCC 15521, Lactobacillus Sake MGB0632 and Lactobacillus Sake MGB0522. Among the Lactobacillus sakei strains of the present specification, ATCC 15521 is deposited with the Korean Cell Line Bank, and MGB0632 and MGB0522 are deposited with the Microorganism and Gene Bank. It can be easily obtained by applying.

In the present specification, the "Lactobacillus sakei" may include Lactobacillus Sake ATCC 15521: Lactobacillus Sake MGB0632: Lactobacillus Sake MGB0522 in a relative weight ratio of 1: 0.5 to 1.5: 0.5 to 1.5 have. Under the weight ratio as described above, the moisturizing and anti-aging effects (wrinkle improvement, elasticity enhancement, etc.) intended for the present specification may be more maximized.

In the present specification, the term "fermented product" is not limited in form and properties, and includes, without limitation, a product obtained during the fermentation process of the mixture or after the fermentation process is completed.

In the present specification, "active ingredient" refers to an ingredient that exhibits a desired activity by itself or can exhibit activity together with a carrier that does not have its own activity.

In one aspect, the present specification is a cosmetic composition for skin moisturizing comprising a fermented product of Lactobacillus sakei of a mixture of horseback riding, dermis, gardenia, oleracea and licorice as an active ingredient, and the Lactobacillus sakei (Lactobacillus sakei) It relates to one or more compositions selected from the group consisting of Lactobacillus Skay ATCC 15521, Lactobacillus Skay MGB0632 and Lactobacillus Skay MGB0522.

In another aspect, the present specification is a pharmaceutical composition for skin moisturizing comprising a fermented product of Lactobacillus sakei of a mixture of horseback riding, dermis, gardenia, oleracea and licorice as an active ingredient, and the Lactobacillus sakei is It relates to one or more compositions selected from the group consisting of Lactobacillus SAK ATCC 15521, Lactobacillus SAK MGB0632 and Lactobacillus SAK MGB0522.

In addition, the present specification is a health food composition for skin moisturizing comprising a fermented product of Lactobacillus sakei of a mixture of horseback riding, dermis, gardenia, oleracea and licorice as an active ingredient from another viewpoint, and the Lactobacillus sakei (Lactobacillus sakei ) relates to a composition that is one or more selected from the group consisting of Lactobacillus SAK ATCC 15521, Lactobacillus SAK MGB0632 and Lactobacillus SAK MGB0522.

The above “moisturizing” may mean increasing skin moisture content or improving skin texture. In addition, since the above active ingredients included in the composition as an aspect of the present invention can rapidly increase the content of hyaluronic acid in the skin, there is also an advantage that it can be used as a composition for preventing or treating diseases such as psoriasis and atopy. .

In another aspect, the present specification is a cosmetic composition for improving or alleviating wrinkles comprising a fermented product of Lactobacillus sakei of a mixture of horseback riding, dermis, gardenia, oleracea and licorice as an active ingredient, and the Lactobacillus sakei (Lactobacillus sakei ) may relate to one or more compositions selected from the group consisting of Lactobacillus SAK ATCC 15521, Lactobacillus SAK MGB0632 and Lactobacillus SAK MGB0522.

In addition, in one aspect, the present specification is a pharmaceutical composition for preventing wrinkle formation comprising a fermented product of Lactobacillus sakei of a mixture of horseback riding, dermis, gardenia, ovary and licorice as an active ingredient, and the Lactobacillus sakei (Lactobacillus sakei ) relates to a composition that is one or more selected from the group consisting of Lactobacillus SAK ATCC 15521, Lactobacillus SAK MGB0632 and Lactobacillus SAK MGB0522.

In another aspect, the present specification is a health food composition for improving wrinkles, comprising a fermented product of Lactobacillus sakei of a mixture of horseback riding, dermis, gardenia, ovary and licorice as an active ingredient, and the Lactobacillus sakei (Lactobacillus sakei ) relates to a composition that is one or more selected from the group consisting of Lactobacillus SAK ATCC 15521, Lactobacillus SAK MGB0632 and Lactobacillus SAK MGB0522.

In addition, in one aspect, the present specification is a cosmetic composition for improving skin elasticity, comprising a fermented product of Lactobacillus sakei of a mixture of horseback riding, dermis, gardenia, oleracea and licorice as an active ingredient, and the Lactobacillus sakei ( Lactobacillus sakei ) includes at least one composition selected from the group consisting of Lactobacillus Sake ATCC 15521, Lactobacillus Sake MGB0632 and Lactobacillus Sake MGB0522.

In addition, the present specification is a pharmaceutical composition for preventing skin elasticity reduction, comprising a fermented product of Lactobacillus sakei of a mixture of horseback riding, dermis, gardenia, oleracea and licorice from another viewpoint as an active ingredient, the Lactobacillus sakei ( Lactobacillus sakei ) relates to one or more compositions selected from the group consisting of Lactobacillus Sake ATCC 15521, Lactobacillus Sake MGB0632 and Lactobacillus Sake MGB0522.

In another aspect, the present specification is also a health food composition for improving skin elasticity, comprising a fermented product of Lactobacillus sakei of a mixture of horseback riding, dermis, gardenia, oleracea and licorice as an active ingredient, the Lactobacillus sakei ( Lactobacillus sakei ) relates to one or more compositions selected from the group consisting of Lactobacillus Sake ATCC 15521, Lactobacillus Sake MGB0632 and Lactobacillus Sake MGB0522.

When the composition as an aspect of the present invention contains a fermented product fermented with one or more strains selected from the group consisting of Lactobacillus SAK ATCC 15521, Lactobacillus SAK MGB0632 and Lactobacillus SAK MGB0522, it has superior moisturizing activity. , It has anti-aging activity (wrinkle improvement, elasticity improvement, etc.), especially in the case of fermented products containing all of the above three strains, exhibits remarkably excellent moisturizing and anti-aging effects.

The above active ingredients included in the composition as an aspect of the present invention can inhibit elastase and collagenase, which are the causes of skin wrinkles and elasticity reduction, and increase the synthesis of procollagen, a precursor of collagen, to reduce skin wrinkles. There is an advantage of being able to perform the effect of improving and enhancing elasticity more actively, so it is possible to more effectively delay skin changes over time related to skin elasticity, wrinkle formation, and the like.

In the composition of one aspect of the present invention, the mixture may contain 2 to 4 horseback riding, 1.5 to 3.5 dermis, 1 to 3 licorice, and 0.5 to 2.5 of gardenia relative to the weight of the five baeja. .

The cosmetic composition, which is one aspect of the present invention, is not particularly limited in its formulation, and may be appropriately selected according to the intended purpose. For example, softening lotion (skin lotion and milk lotion), nutritional lotion, essence, nutrition cream, massage cream, pack, gel, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, pack, powder, It may be prepared in one or more formulations selected from the group consisting of body lotion, body cream, body oil, and body essence, but is not limited thereto. In addition, the cosmetic composition may include use as an external preparation for skin in the form of an ointment, patch, or the like.

The cosmetic composition according to the present invention can be provided in any formulation suitable for topical application. For example, it may be provided in the form of a solution, an emulsion obtained by dispersing an oil phase in an aqueous phase, an emulsion obtained by dispersing an aqueous phase in an oil phase, a suspension, a solid, a gel, a powder, a paste, a foam, or an aerosol composition. Compositions of such formulations can be prepared according to conventional methods in the art.

The cosmetic composition according to the present invention may include other ingredients that can give a synergistic effect to the main effect, preferably within a range that does not impair the main effect in addition to the above-described substances. In addition, the cosmetic composition according to the present invention may further include a moisturizing agent, an emollient agent, an ultraviolet absorber, a preservative, a disinfectant, an antioxidant, a pH adjuster, an organic and inorganic pigment, a fragrance, a cooling agent or a limiting agent. The blending amount of the ingredients can be easily selected by a person skilled in the art within a range that does not impair the object and effect of the present invention, and the blending amount may be 0.01 to 5% by weight, specifically 0.01 to 3% by weight, based on the total weight of the composition. have.

In the pharmaceutical composition according to another aspect of the present invention, when the fermented product of the lactic acid bacteria of the mixture is applied to a pharmaceutical product, the composition is used as an active ingredient and a commercially available inorganic or organic carrier is added to form a solid, semi-solid, or liquid form. It can be formulated as an administration or parenteral administration.

The preparations for oral administration include tablets, pills, granules, capsules, powders, fine granules, powders, emulsions, syrups, pellets, and the like. In addition, the preparations for parenteral administration include injections, drops, ointments, lotions, sprays, suspensions, emulsions, and suppositories. In order to formulate the active ingredient of the present invention, it can be easily formulated if carried out according to conventional methods, and surfactants, excipients, coloring agents, spices, preservatives, stabilizers, buffers, suspensions, and other commonly used auxiliary agents can be appropriately used.

The pharmaceutical composition according to the present invention may be administered orally, parenteral, rectal, topical, transdermal, intravenous, intramuscular, intraperitoneal, subcutaneous, and the like.

The active ingredient of the pharmaceutical composition of the present invention will vary depending on the age, sex, weight, pathological condition and severity of the subject to be administered, the route of administration, or the judgment of the prescriber. Determination of the applied amount based on these factors is within the level of those skilled in the art, and its daily dose is, for example, 0.1 mg/kg/day to 100 mg/kg/day, more specifically 5 mg/kg/day to 50 mg/kg. /It can be work, but is not limited thereto.

The present invention is another aspect of the health food composition provides various types of food additives or functional foods. It can be processed into leached tea, liquid tea, beverages, fermented milk, cheese, yogurt, juice, probiotics, health supplements, etc. including the composition, and can be used in the form of various other food additives.

In one embodiment, the composition may contain other components that can give a synergistic effect to the main effect within a range that does not impair the main effect targeted by the present invention. For example, additives such as fragrances, pigments, disinfectants, antioxidants, preservatives, moisturizers, thickeners, inorganic salts, emulsifiers, and synthetic polymer substances may be further included in order to improve physical properties. In addition, auxiliary components such as water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, and seaweed extract may be further included. The above ingredients may be appropriately selected and blended by a person skilled in the art according to the formulation or purpose of use, and the amount added may be selected within a range not impairing the object and effect of the present invention. For example, the amount of the components added may be in the range of 0.01 to 5% by weight, more specifically 0.01 to 3% by weight, based on the total weight of the composition.

The formulation of the composition according to the present invention may be in various forms such as solutions, emulsions, viscous mixtures, tablets, powders, etc., which may be administered by various methods such as simple drinking, injection administration, spray method, or squeeze method.

Hereinafter, the present invention will be described in more detail through examples. These examples are for illustrative purposes only, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not construed as being limited by these examples.

[Example] Preparation of fermented lactic acid bacteria

Horse riding, dermis, licorice, gardenia, and ovary were mixed in a ratio of 30:25:20:15:10, and then sterilized. Distilled water was added as much as 10 times the volume of the raw material, extracted for 4 hours, heated at 80, and then aged for 12 hours to obtain a mixture.

Put 5% of the above mixture in relation to the minimum medium weight in the lactic acid bacteria minimum medium (2% whole milk, based on 1L), ^{and inoculate each of Lactobacillus sakei of the composition shown in Table 1 at 1.8*10 8} CFU/ml Incubated for 30 to 48 hours. The culture result was filtered under reduced pressure and concentrated under reduced pressure, and then freeze-dried to obtain a powder, which was used in the following experimental examples.

Strain Mixing ratio Indications in the following drawings Example 1 ATCC 15521 #One Example 2 MGB0632 #2 Example 3 MGB0522 #3 Example 4 ATCC 15521, MGB0632 1:1 #4 Example 5 ATCC 15521, MGB0522 1:1 #5 Example 6 MGB0632, MGB0522 1:1 #6 Example 7 ATCC 15521, MGB0632, MGB0522 1:1:1 #7

[Experimental Example 1] Antioxidant Activity of Fermented Lactobacillus

In order to examine the antioxidant effect of each of the fermented lactic acid bacteria prepared in Examples 1 to 7, the free radical scavenging ability was measured by DPPH (1,1-diphenyl-2-picryl-hydrazyl) assay and ABTS assay. L-ascobic acid (L-A), an antioxidant known in the art, was used as a positive control. Each of the experimental groups was tested using 100 μg/ml, 200 μg/ml, and 400 μg/ml, respectively, and as a result, FIGS. 1 and 2 were obtained.

1 and 2 thus obtained, the active ingredient of the composition of the present invention, specifically, Lactobacillus SAK ATCC 15521, Lactobacillus SAK MGB0632 and Lactobacillus SAK MGB0522 fermented with one or more strains selected from the group consisting of The obtained fermented product (Examples 1 to 3, # of FIGS. 1 and 2) containing only one strain of the fermented product (corresponding to Examples 4 to 7, #4 to #7 of FIGS. 1 and 2) Corresponds to 1 to #3). In addition, fermentation products obtained using all three strains including Lactobacillus SAK ATCC 15521, Lactobacillus SAK MGB0632 and Lactobacillus SAK MGB0522 (corresponding to #7 in Example 7, FIGS. 1 and 2) It was confirmed that the fermented product obtained using only two strains of the above strains (corresponding to Examples 4 to 6 and #4 to #6 in FIGS. 1 and 2) had better antioxidant activity.

[Experimental Example 2] Confirmation of the presence or absence of cytotoxicity of fermented lactic acid bacteria

To confirm cytotoxicity, B16/F1 (Mouse/Melanoma), CCD-986sk (Human/Fibroblast), and Hacat (Human/Keratinocyte) cells distributed from the Korean cell line bank were used.

In DMEM (Lonza) medium containing 1% penicillin/streptomycin (GIBCO) and 10% fetal bovine serum (GIBCO), each of the cells obtained by Examples 4 to 7 The sample was treated and incubated in a 37° C., 5% CO 2 thermostat. Thereafter, cytotoxicity was confirmed using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay in each of the cultured cells, and are shown in FIGS. 6 and 7. MTT is a pale yellow substrate that reacts with enzymes in the mitochondria of living cells to form formazan, which is dissolved in dimethyl sulfoxide (DMSO) and the absorbance of 570 nm is measured to measure cell viability.

As a result, as can be seen in FIGS. 3 to 5, the fermented product of lactic acid bacteria, which is an active ingredient of the composition of the present invention, did not show cytotoxicity.

[Experimental Example 3] Elastase Inhibitory Activity of Fermented Lactic Acid Bacteria

Each fermented product obtained in Examples 4 to 7 was taken at 100 μg/ml, 200 μg/ml, and 400 μg/ml in a 96-well plate, and a 2.5 U/ml porcine pancreas elastase solution dissolved in 50 mM Tris-HCl buffer (pH 8.6) was prepared. 40 µl was added. After adding n-succinyl-(L-Ala)3-pnitroanilide (0.5 mg/ml) dissolved in 50mM Tris-HCl buffer (pH 8.6) as a substrate, reacting for 30 minutes, and then standing on ice for 5 minutes to stop the reaction. Absorbance was measured at 410 nm. Ursolic acid (U-A) was used as a positive control.

As a result, as can be seen in Figure 6, a fermentation product obtained using all three strains including Lactobacillus SAK ATCC 15521, Lactobacillus SAK MGB0632 and Lactobacillus SAK MGB0522 (Example 7, FIG. 1 And #7 of FIG. 2) is an elastase that is far superior to the fermented product obtained using only two strains of the above strains (corresponding to Examples 4 to 6 and #4 to #6 of FIGS. 1 and 2). It was confirmed that it showed an activity inhibitory ability.

[Experimental Example 4] Collagenase Inhibitory Activity of Fermented Lactobacillus

Add 4mM CaCl2 to 0.1M Tris-HCl buffer (pH7.5) and dissolve 4-phenylazobenzyloxycarbonyl-Pro-Leu-Gly-Pro-Arg (0.3mg/ml) in 0.25ml of the obtained substrate solution. Each of the fermented products obtained in Examples 4 to 7 was added to each of 100 μg/ml, 200 μg/ml, and 400 μg/ml, and then 0.15 ml of collagenase (0.2 mg/ml) was added and left at room temperature for 20 minutes. Tannic acid was used as a positive control. After adding 0.5 ml of 6% citric acid to stop the reaction of collagenase, 1.5 ml of ethyl acetate was added and mixed well, and the absorbance was measured at 320 nm. Collagenase is an enzyme that breaks down collagen, so inhibiting its activity can inhibit collagen breakdown, so it is effective in improving skin elasticity and wrinkles.

As a result, as can be seen in Figure 7, the fermented product obtained by using all three strains including Lactobacillus SAK ATCC 15521, Lactobacillus SAK MGB0632 and Lactobacillus SAK MGB0522 (Example 7, FIG. 8 It was confirmed that the collagenase inhibitory activity was superior to the fermented product obtained by using only two strains of the above strains (corresponding to #7 of) (Examples 4 to 6 and #4 to #6 of FIG. 8). .

[Experimental Example 5] Procollagen synthesis ability of fermented lactic acid bacteria

In a serum-free condition, the fermented products of Examples 4 to 7 were treated with each of 25 μg/ml, 50 μg/ml, and 100 μg/ml to CCD-986sk (Human/Fibroblast) cells distributed from the Korean cell line bank and cultured for 72 hours. . The experimental group not treated with the above fermentation was used as a control group (Non). The content of procollagen synthesized according to each sample was measured with a procollagen ELISA Kit (Elabscience, USA).

As a result, as can be seen in Figure 8, the fermented product obtained using all three strains including Lactobacillus SAK ATCC 15521, Lactobacillus SAK MGB0632 and Lactobacillus SAK MGB0522 (Example 7, FIG. 9 It was confirmed that the procollagen synthesis ability was superior to the fermented product obtained using only two strains of the above strains (corresponding to #7 of) (Examples 4 to 6, corresponding to #4 to #6 of FIG. 9).

[Experimental Example 6] Increased hyaluronic acid production by fermented lactic acid bacteria

HaCat cells (Korea Cell Line Bank) were seeded in a 6 well plate at a concentration of ^{2 x 10 5 cells/ml.} After 24 hours, after washing twice with serum-free DMEM (Lonza DMEM with 4.5g/L Glucose & L-Glutamine 500ml part number: 12-604F / country of manufacture: USA), the serum-free DMEM was replaced and the fermented products of Examples 4 to 7 Were treated at 100 μg/ml increments. The experimental group not treated with the above fermentation was used as a control group (Non). After 24 hours, it was centrifuged at 15,000 xg for 5 minutes (Smart-R17/manufacturer: Hanil Science/manufacturer: Korea), and the supernatant was removed and used. After the supernatant was added to the plate coated with the hyaluronic acid antibody and reacted, a calibration curve was drawn to measure the result, and FIG. 10 was obtained.

As a result, the fermented product obtained by using all three strains including Lactobacillus SAK ATCC 15521, Lactobacillus SAK MGB0632 and Lactobacillus SAK MGB0522 (corresponding to Example 7, #7 of FIG. 9) above It was confirmed that the fermented product obtained by using only two strains of the strains (Examples 4 to 6, corresponding to #4 to #6 in FIG. 9) showed a far superior moisturizing effect.

[Experimental Example 7] Clinical evaluation of anti-wrinkle effect of fermented lactic acid bacteria

22 subjects were selected to self-apply the cream having the same composition as Formulation Example 2 disclosed in the present specification to the skin for 4 weeks, and then the wrinkle improvement effect was observed before application, 2 weeks after application, and 4 weeks after application. The specific wrinkle improvement effect was obtained by using the Antera 3D device to obtain an indentation index at the test site and compared them.

As a result, as shown in FIG. 10, it was confirmed that the wrinkles were remarkably relieved as the application period increased.

Hereinafter, an example of the formulation of the composition according to the present invention will be described, but this can be applied to various formulations, which is intended to be described only in detail, not to limit the present invention.

[Formulation Example 1] Soft Capsule

The fermented product of Example 7 150 mg, palm oil 2 mg, hydrogenated palm oil 8 mg, sulfur lead 4 mg, and lecithin 6 mg were mixed, and 400 mg per capsule was filled according to a conventional method to prepare a soft capsule.

[Formulation Example 2] Tablet

150 mg of the fermented product of Example 7, 100 mg of glucose, 50 mg of red ginseng extract, 96 mg of starch, and 4 mg of magnesium stearate were mixed, and 40 mg of 30% ethanol was added to form granules, followed by drying at 60°C and a tableting machine. It was compressed into tablets using.

[Formulation Example 3] Granules

150 mg of the fermented product of Example 7, 100 mg of glucose, 50 mg of red ginseng extract, and 600 mg of starch were mixed to form granules, and 100 mg of 30% ethanol was added to form granules, followed by drying at 60°C to form granules. Filled. The final weight of the contents was 1 g.

[Formulation Example 4] Drinks

150 mg of the fermented product of Example 7, 10 g of glucose, 50 mg of red ginseng extract, 2 g of citric acid, and 187.8 g of purified water were mixed and filled into a bottle. The final volume of the contents was 200 ml.

[Formulation Example 5] Preparation of healthy food

Fermented product of Example 7............1000 mg

Vitamin mixture

Vitamin A acetate............70 ㎍

Vitamin E ........................................... 1.0 mg

Vitamin B1................................0.13 mg

Vitamin B2 ...............................0.15 mg

Vitamin B6........................................... 0.5 ㎎

Vitamin B12.......................................... 0.2 ㎍

Vitamin C............................................. 10 ㎎

Biotin.................. 10 ㎍

Nicotinic acid amide......................................1.7 mg

Folic acid................................................. 50 ㎍

Calcium pantothenate............ 0.5 mg

Mineral mixture

Ferrous sulfate.......................................... 1.75 mg

Zinc oxide...........................................0.82 mg

Magnesium carbonate.......................................25.3 mg

Potassium Phosphate Monobasic.........................................15 ㎎

Dicalcium Phosphate..........................................55 ㎎

Potassium citrate...........................................90 mg

Calcium carbonate............................................100 mg

Magnesium chloride.......................................24.8 mg

The composition ratio of the vitamin and mineral mixture is relatively suitable for health food, but it may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. , To prepare granules, and can be used for preparing a health food composition according to a conventional method.

[Formulation Example 6] Preparation of healthy beverage

Fermented product of Example 7............1000 mg

Citric acid..................1000 ㎎

Oligosaccharide............................100 g

Plum concentrate............................................2 g

Taurine...................................... 1 g

Add purified water to the whole...............900 ml

After mixing the above ingredients according to a conventional health drink manufacturing method, stirring and heating at 85° C. for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 liter container, sealed and sterilized, and then stored in a refrigerator. It is used in the manufacture of health drink composition.

The composition ratio is composed of ingredients suitable for a relatively preferred beverage in a preferred embodiment, but the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as the demand class, the country of demand, and the intended use. Those of ordinary skill in the technical field to which the present invention belongs will be able to perform various applications and modifications within the scope of the present invention based on the above contents.

[Formulation Example 1] Lotion

Ingredients weight% Fermented product of Example 7 3.00 L-ascorbic acid-2-phosphate magnesium salt 1.00 Water-soluble collagen (1% aqueous solution) 1.00 Sodium citrate 0.10 Citric acid 0.05 White Oyster Extract 0.20 1,3-butylene glycol 3.00 Purified water to 100

[Formulation Example 2] Cream

Ingredients weight% Fermented product of Example 7 1.00 Polyethylene glycol monostearate 2.00 Self-emulsifying glycerin monostearate 5.00 Cetyl alcohol 4.00 Squalene 6.00 Tri2-ethylhexane glyceryl 6.00 Sphingoglycolipid 1.00 1,3-butylene glycol 7.00 Purified water to 100

[Formulation Example 3] Pack

Ingredients weight% Fermented product of Example 7 5.00 Polyvinyl alcohol 13.00 L-ascorbic acid-2-phosphate magnesium salt 1.00 Lauroylhydroxyproline 1.00 Water-soluble collagen (1% aqueous solution) 2.00 1,3-butylene glycol 3.00 ethanol 5.00 Purified water to 100

[Formulation Example 4] Essence

Ingredients weight% Fermented product of Example 7 2.00 Hydroxyethylene Cellulose (2% aqueous solution) 12.00 Xanthan gum (2% aqueous solution) 2.00 1,3-butylene glycol 6.00 Dark glycerin 4.00 Sodium hyaluronate (1% aqueous solution) 5.00 Purified water to 100

[Formulation Example 5] Ointment

Ingredients Content (% by weight) Fermented product of Example 7 0.1 glycerin 8.0 Butylene glycol 4.0 Floating paraffin 15.0 Beta Glucan 7.0 Carbomer 0.1 Caprylic/Capric Triglyceride 3.0 Squalane 1.0 Cetearyl glucoside 1.5 Sorbitan stearate 0.4 Cetearyl alcohol 1.0 Beeswax 4.0 Preservatives, colors, flavors Appropriate amount Purified water Balance

As described above, specific parts of the present invention have been described in detail, and it is obvious that these specific techniques are only preferred embodiments for those of ordinary skill in the art, and the scope of the present invention is not limited thereby. something to do. Accordingly, it will be said that the substantial scope of the present invention is defined by the appended claims and their equivalents.

Claims (5)

delete A cosmetic composition for improving wrinkles containing a fermented product of Lactobacillus sakei of horseback riding, dermis, gardenia, oleracea and licorice as an active ingredient,
The mixture comprises horseback riding 2 to 4, dermis 1.5 to 3.5, licorice 1 to 3, and gardenia in a relative weight ratio of 0.5 to 2.5,
The Lactobacillus sakei (Lactobacillus sakei) is a cosmetic composition for wrinkle improvement comprising Lactobacillus Sake ATCC 15521: Lactobacillus Sake MGB0632: Lactobacillus Sake MGB0522 in a relative weight ratio of 1: 1: 1. A cosmetic composition for improving skin elasticity comprising a fermented product of Lactobacillus sakei of a mixture of horseback riding, dermis, gardenia, oleracea and licorice as an active ingredient,
The mixture comprises horseback riding 2 to 4, dermis 1.5 to 3.5, licorice 1 to 3, and gardenia in a relative weight ratio of 0.5 to 2.5,
The Lactobacillus sakei (Lactobacillus sakei) is a cosmetic composition for improving skin elasticity comprising Lactobacillus Sake ATCC 15521: Lactobacillus Sake MGB0632: Lactobacillus Sake MGB0522 in a relative weight ratio of 1: 1: 1. delete delete

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