KR101587077B1 - Composition containing fermentated opuntia humifusa showing biological activity of skin - Google Patents

Abstract

The present invention relates to a composition comprising a fermented product of perennial plant.

Description

Technical Field [0001] The present invention relates to a composition comprising a fermented product having a skin physiological activity,

The present invention relates to a composition comprising a fermented product of perennial plant.

 Recently, as the level of living has improved, the interest of modern people in skin beauty is increasing. In particular, there is a growing interest in preventing skin aging in order to have a face that looks younger than age.

As the largest organ of the human body, which occupies about 16% of the total human body volume, the skin is directly contacted with the external environment, and it is important to protect the human body from many harmful factors that intrude into the human body such as temperature, humidity and ultraviolet rays. It acts as a protective film. However, with aging, skin cells are damaged due to various pollutants, strong ultraviolet rays, stress and nutritional deficiency, and regeneration is not actively performed, resulting in wrinkles on the skin and loss of elasticity.

Conventionally, natural plants have been used as they are in the form of extracts and used as formulations for cosmetics and the like in order to suppress skin aging. However, no formulation or the like including a fermentation product obtained by fermenting a specific natural plant has been reported, nor has a fermentation product containing a specific compound been reported.

Quan, TH., He, TY., Kang, S., Voorhees, JJ., & Fisher, GJ. (2004). Solar ultraviolet irradiation reduces collagen in photoaged human skin by blocking transforming growth factor-beta type II receptor / Smad signaling. Am J Pathol, 165 (3), 741-51

It is an object of the present invention to provide a composition comprising a natural fermenting substance comprising a specific compound and a composition comprising a natural fermenting substance having a novel use.

In order to accomplish the above object, one aspect of the present invention is a composition including a fermented product of Paeoniae sp., Wherein the Paeoniae fermented product is selected from the group consisting of caffeic acid, p-coumaric acid, ferulic acid, O-glucoside, isorhamnetin 3-O-rutinoside, and isorhamnetin 3-O-glucoside. , ≪ / RTI >

One aspect of the present invention also provides a composition for suppressing or improving skin photoaging, which comprises a fermented product of Baekjanggi as an active ingredient.

In addition, one aspect of the present invention provides a composition for skin moisturizing, which comprises a fermented product of Baekjanggi as an active ingredient.

One aspect of the present invention provides a new indication that a fermented product of Paeoniae contains a specific indicator substance that did not exist in Baekseoncho. In addition, there is a significance in increasing the possibility of using the fermented product of Baekjangwon for various purposes.

In addition, the composition of one aspect of the present invention can increase the production of TGF-β and decrease the production of MMP, including fermented products of the perennial plant having an effect superior to that of the perennial plant, It can be useful. One aspect of the present invention is that the composition can also increase the production of filla greens, including fermented products of the perennial plant having an effect superior to that of the perennial plant, and thus can be usefully used for moisturizing the skin.

In addition, since the fermented product is produced as a naturally derived ingredient, it has little side effects even when ingested or applied, and is natural-friendly.

FIG. 1 is a BPI chromatogram (UPLC-QTOF-MS / MS) of a methanol extract of methanol extract of Baicin Seeds and a methanol extract of Baicin Seeds fermented product.
FIG. 2 shows the results of analysis of the chemical structure of the indicator substances contained in the methanol extracts of methanol extracts of Paeonia spp. And Paeoniae spp. (A: Paekcheon, b: Perennial fermentation products).
FIG. 3 is a graph of PCA / scores (FO: perennial fermentation, EO: perennial) obtained by analyzing the methanol extract of the perennial plant and the methanol extract of the perennial plant fermented by Pallet analysis of UPLC-QTOF-MS / MS.
FIG. 4 is a graph showing changes in the concentration of TGF-β after treatment with UVB-irradiated fibroblast culture model of each of the perennial and perennial fermentations (FO: perennial fermentation, EO: perennial).
FIG. 5 is a graph showing changes in MMP-1 concentration (FO: perennial fermentation, EO: perennial) in a fibroblast culture model irradiated with UVB.
FIG. 6 is a graph (FO: perennial fermentation product, EO: perennial) in which a water-content of transdermal tissue is measured by treating each of the perennial plant and perennial plant fermented product with a UVB-irradiated mouse model.
Fig. 7 shows the results of treatment of each of the perennial and perennial fermented products in a mouse model irradiated with UVB. (FO: fermented water, EO: perennial) of trichrome staining.
FIG. 8 shows the result of treatment of each of the perennial and perennial fermentations with a UVB-irradiated mouse model, and measurement of the transdermal water content by Western blotting (FO: ferret of perennial plant, EO: perennial plant).

In one aspect, the present invention is a composition comprising a fermented product of Paeoniae sp., Wherein the Paeoniae fermented product is selected from the group consisting of caffeic acid, p-coumaric acid, ferulic acid, Wherein the composition comprises at least one compound selected from the group consisting of isorhamnetin 3-O-glucoside, 3-O-rutinoside and isorhamnetin 3-O-glucoside.

The term " Opuntia " humifusa "is a plant belonging to the genus Opuntia in the cactus. It is a native plant of Jeju Island and grows widely in soil which is relatively difficult to grow, and it is cultivated widely in difficult places. Fruits are very rich in dietary fiber, while calories are very low, with 85% moisture, 10-15% carbohydrates, 6-8% glucose and fructose, and vitamins. In the present specification, the perennial herb extract may be obtained without limitation from all parts of the plant including the fruit, root, stem, and leaf of the perennial plant, but may be obtained specifically from the fruit.

In the present specification, the "fermented product of perennial plant" can be obtained by fermenting the perennial plant extract or the perennial plant itself. The perennial herb extract may be obtained by cooling, heating and filtering at room temperature by a conventional method known in the art to obtain a liquid product, or additionally, the solvent may be evaporated, spray dried or freeze-dried, and there is no limitation in the production method thereof. Even when fermenting the perennial plant itself, it can be used without limitation if it contains the solution or powder obtained from the perennial plant or the perennial plant, for example, making the perennial plant powder or using the perennial plant liquid. The perennial fermentation product may be obtained by inoculating two or more kinds of yeast into the perennial herb extract, but is not limited thereto. In the present specification, 'to inoculate two or more kinds of yeast and ferment the perennial herb extract' means to ferment the perennial herb extract by using a culture solution obtained by culturing two or more kinds of yeast, or culturing two or more kinds of yeast together And using the obtained culture medium for the fermentation of the perennial herb extract. However, the present invention is not limited thereto, and there is no limitation as long as it is obvious to a person skilled in the art. As used herein, the yeast may be Aureobasidium sp. And Candida sp., And more specifically, Aureobasidium pullulans and Candida utilis SY8-M2-1 ( (Candida utilis SY8-M2-1), more specifically Aureobasidium pullulans (KCCM 12017) and Candida utilis SY8-M2-1 (Candida utilis SY8-M2-1, KFCC11487P) have. Wherein the fermentation step comprises treating at least 5 parts by weight of a mixed culture of Aureobasidium pullulans and Candida utyl SY8-M2-1 at a temperature of 30 ° C to 40 ° C for 15 to 30 hours when the total weight of the extract is 100 parts by weight, Lt; / RTI > for a period of time. The 'mixed culture solution' may include a culture solution obtained by culturing two or more yeast cultures, or a culture solution obtained by culturing two or more yeasts together. However, the culture solution is not limited to those described in the art, none.

The above-mentioned compounds were found for the first time that the present inventors were present in the fermentation product of Baekryeon.

Specifically, in the composition according to one aspect of the present invention, the fermented product of perennial plant comprises 0.005% to 0.006% by weight of caffeic acid relative to the total weight of fermented product perennial plant; 0.004% to 0.005% by weight of p-coumaric acid; 0.01 to 0.02% by weight of ferric acid; 0.002% to 0.003% by weight of isohamnetine 3-O-rutinoside; Or isohamnetine 3-O-glucoside in an amount of 0.001 wt% to 0.002 wt%.

In one composition of the present invention, the perennial fermentation product is selected from the group consisting of Aureobasidium sp.) and Candida genus (Candida sp.) may be water yeast fermentation.

In one aspect of the present invention, there is provided a composition comprising the Aureobasidium The yeast may be Aureobasidium pullulans , and the Candida sp. yeast may be Candida utilis .

In one aspect of the invention, the composition may be for inhibiting or improving skin photoaging. In a composition according to one aspect of the present invention, the composition may be for treatment of skin photoaging, and may also be for alleviation of skin photoaging.

In the present specification, the term " inhibition or improvement of aging of skin photo-aging "refers to inhibition of aging of skin cells to promote aging slowly or promote regeneration of new cells capable of replacing aged skin, It can mean improvement. In one aspect of the present invention, photoexposure due to ultraviolet rays is progressed slowly, or photoaging phenomenon can be suppressed or photoaging phenomenon can be suppressed. More specifically, in a composition that is an aspect of the present invention, the perennial fermentation product may increase the production of TGF-ss or reduce the production of MMPs. In a composition according to one aspect of the present invention, the perennial fermentation product may increase the production of TGF-beta itself, increase the activity of TGF-beta, or inhibit the activity of TGF- (up-stream) proteins, enzymes and the like. In a composition according to one aspect of the present invention, the fermented product of the perennial plant may reduce the production of MMP itself, increase the activity of MMP, and may inhibit the production of MMP or an up-stream Proteins, enzymes, etc. can be controlled and controlled. In addition, there is no limitation on the kind of the MMP in the composition which is one aspect of the present invention.

In one aspect of the invention, the composition may be for skin moisturizing.

As used herein, the term "skin moisturizing" may mean increasing the amount of moisture remaining in the transdermal or dermis of the skin, or preventing the transdermal or dermis from emitting moisture, thereby increasing the moisture content of the skin cells. Specifically, the fermented product of Paeoniae may increase the production of fillagrin to increase moisturization of the skin, but the present invention is not limited thereto. In the composition according to one aspect of the present invention, the fermented product of the perennial plant may increase the production of the pillared green, increase the activity of the pillared green, increase the activity of the up- protein, enzyme, etc., of the protein.

The present invention relates to a composition for inhibiting or improving skin photoaging, which comprises a fermented product of perennial plant as an active ingredient from another viewpoint.

In one composition of the present invention, the perennial fermentation product is selected from the group consisting of Aureobasidium sp.) and Candida genus (Candida sp.) may be water yeast fermentation. More specifically, the above-mentioned Aureobasidium The yeast may be a yeast strain selected from the group consisting of Aureobasidium pullulans), and the genus Candida (Candida sp.) yeast Candida utility less (Candida utilis .

In a composition that is an aspect of the present invention, the perennial fermentation product may increase TGF-beta production and decrease the production of MMP.

In one aspect of the present invention, the composition may comprise 1.0 to 5.0% by weight of the fermented product of Paeoniae, based on the total weight of the composition. When the composition contains the perennial fermentation product in the weight ratio range, it can effectively increase TGF-beta production, exerts an effect of reducing the production of MMP, and does not affect the content of other constituents. In view of the above, the composition may comprise 1.2 to 4.8 wt.%, 1.4 to 4.6 wt.%, 1.6 to 4.4 wt.%, 1.8 to 4.2 wt.% Or 2.0 to 4.0 wt.% Of the perennial fermentation product have.

The present invention relates to a composition for skin moisturizing which comprises a fermented product of pale herbaceousum as an active ingredient from another viewpoint.

In one aspect of the present invention, the perennial fermentation product may be a yeast fermentation product of Aureobasidium sp. And Candida sp. More specifically, the Aureobasidium sp. Yeast may be Aureobasidium pullulans, and the Candida sp. Yeast may be Candida utilis.

In a composition that is an aspect of the present invention, the perennial fermentation product may increase the production of filla green.

In one aspect of the present invention, the composition may comprise 1.0 to 5.0% by weight of the fermented product of Paeoniae, based on the total weight of the composition. When the composition contains the perennial fermentation product in the weight ratio range, the effect of increasing the production of pillared green is excellent, and the content of the other components is not affected. In view of the above, the composition may comprise 1.2 to 4.8 wt.%, 1.4 to 4.6 wt.%, 1.6 to 4.4 wt.%, 1.8 to 4.2 wt.% Or 2.0 to 4.0 wt.% Of the perennial fermentation product have.

One aspect of the present invention is a cosmetic composition.

The cosmetic composition may be, for example, a basic cosmetic composition, a makeup cosmetic composition, a hair cosmetic composition, a body cosmetic composition or the like, and the formulation is not particularly limited and may be appropriately selected according to the purpose.

For example, the cosmetic composition can be formulated into a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, But is not limited thereto. More specifically, the present invention relates to a cosmetic composition, which comprises at least one of a softening agent, a nutritional lotion, a lotion, a body lotion, a nutritional cream, a massage cream, a moisturizing cream, a hand cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, Shampoos, rinses, body cleansers, toothpastes or mouthwashes, hair tonics, cosmetics, cosmetics, cosmetics, cosmetics, cosmetics, Gel or mousse, or a hair cosmetic composition such as a hair dye or a hair dye.

The cosmetic composition contains a cosmetically acceptable medium or base. It may be in any form suitable for topical application, for example as a solution, a gel, a solid or a paste anhydrous product, an emulsion obtained by dispersing an oil phase in water, a suspension, a microemulsion, a microcapsule, a microgranule or an ionic form (liposome) and / In the form of a non-ionic follicle dispersing agent, or in the form of creams, skins, lotions, powders, ointments, sprays or conical sticks. These compositions may be prepared according to conventional methods in the art.

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

In the case where the formulation of the present invention is a suspension, a carrier such as 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, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

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

In the case where the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. Especially, in the case of a spray, a mixture of chlorofluorohydrocarbons, propane / Propane or dimethyl ether.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, or ethoxylated glycerol fatty acid esters.

In one embodiment of the present invention, the cosmetic composition may further contain an enhancer. The thickening agent included in the cosmetic composition of the present invention may be selected from the group consisting of methylcellulose, carboxymethylcellulose, carboxymethylhydroxyguanine, hydroxymethylcellulose, hydroxyethylcellulose, carboxyvinyl polymer, polyquaternium, cetearyl alcohol, Carrageenan, and the like. Among them, at least one of carboxymethyl cellulose, carboxyvinyl polymer, and polyquaternium can be used, and most preferably, it can be a carboxyvinyl polymer.

In one embodiment of the present invention, the cosmetic composition may contain various bases and additives as appropriate, and the kind and amount of these components can be easily selected by the inventors. The composition may contain an additive that is acceptable according to need, and may further include components such as preservatives, pigments, and additives customary in the art.

The preservative may specifically be phenoxyethanol or 1,2-hexanediol, and the perfume may be an artificial perfume or the like.

And, in one embodiment of the present invention, the cosmetic composition may comprise a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymeric polysaccharides, sphingolipids and seaweed extracts. Examples of the compounding ingredients that may be added include organic solvents such as a preservative component, a moisturizer, an emollient, a surfactant, an organic and inorganic pigment, an organic powder, an ultraviolet absorbent, a preservative, a bactericide, an antioxidant, a plant extract, a pH adjuster, A blood circulation accelerator, a cold agent, an antiperspirant agent, and purified water.

The components to be added in addition to these components are not limited thereto, and any of the above components can be compounded within a range that does not impair the objects and effects of the present invention.

A composition that is an aspect of the invention may also be a pharmaceutical composition.

In one embodiment of the present invention, the pharmaceutical composition may be formulated into a parenteral administration form in solid, semi-solid or liquid form by adding a commercially available inorganic or organic carrier, excipient and diluent to the culture medium of the dermal epidermal stem cell as an active ingredient . The preparation for parenteral administration may be a transdermal dosage form selected from the group consisting of drops, ointments, lotions, gels, creams, patches, sprays, suspensions, and emulsions.

Examples of carriers, excipients and diluents that can be included in the composition include lactose, dextrose, sucrose, oligosaccharide, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

In the composition according to each formulation, other components other than the composition of the present invention may be mixed and selected without difficulty by those skilled in the art according to the formulation or use purpose of other external preparation for skin, etc. In this case, A synergistic effect can occur.

Further, when the composition according to the present invention is used as a pharmaceutical composition, it may further contain a preservative, a stabilizer, a wetting agent or an emulsifying accelerator, a pharmaceutical adjuvant such as a salt or buffer for controlling the osmotic pressure, and other therapeutically useful substances And can be formulated into various oral or parenteral dosage forms according to conventional methods.

It should be understood that the actual dosage of the active ingredient should be determined in light of various relevant factors such as the severity of the symptoms, the route of administration selected, the age, sex, weight and health status of the subject.

Generally, the dosage of active ingredient is in the range of 0.001 mg / kg / day to about 2000 mg / kg / day. A more preferred dosage is 0.5 mg / kg / day to 2.5 mg / kg / day. External administration may be administered once a day or divided into several doses.

Hereinafter, the present invention will be described more specifically with reference to the following examples. However, the following examples are provided for illustrative purposes only in order to facilitate understanding of the present invention, and the scope and scope of the present invention are not limited thereto.

[ Comparative Example  1] Preparation of extracts (powders)

5 kg of purified water is added to the crushed white birch seeds (1 kg), and the mixture is subjected to hot water extraction while stirring at 80 ° C for 1 hour or more. The extract was filtered using a filter press (IJ Engineering / Korea) and then pulverized using a powder drier (Eugene Tech / Korea).

[ Example  1] Fermentation product  Preparation of extract

[ Example  1-1] Aureobasidium Pullulanth  ( Aureobasidium pullulans ) Culture

Aureobasidium < RTI ID = 0.0 > pullulans , KCCM 12017) were purchased from the Korean Society of Bacterial Kidney. Aureobasidium pullulans was subcultured in YM agar medium at 30 DEG C for 24 hours and then stored in a refrigerator for seed culture. One platinum sample was inoculated in a test tube containing 5 ml of the seed culture medium in the refrigerated Aureobasidium pullulans (KCCM 12017) slant and seeded.

[ Example  1-2] Candida Utility SY8 - M2 -One( Candida utilis SY8 - M2 -1)

Candida utyl SY8-M2-1 was subcultured in YM agar medium at 30 ° C for 24 hours, and then stored in a refrigerator for seed culture. One platinum sample was inoculated into a test tube containing 5 ml of the seed culture medium from the slant of Candida utilis SY8-M2-1 ( Candida utilis SY8-M2-1), which was stored in the refrigerator.

[ Example  1-3] Aureobasidium Pullulanth  ( Aureobasidium pullulans ) And Candida Utility SY8 - M2 -One( Candida utilis SY8 - M2 -1) complex culture

Examples 1-1 Aureobasidium pullulans (Aureobasidium pullulans) as in Candida utility less SY8-M2-1 (Candida obtained in Examples 1-2 obtained by the utilis SY8-M2-1) seed culture.

[ Example  1-4] Fermented  Produce

5 kg of purified water was added to the crushed white perennials (1 kg), and the mixture was subjected to hot water extraction with stirring at 80 캜 for 1 hour or more. Cellulase corresponding to 0.1% (w / w) of the weight (1 kg) of the perennial flower was added to the extract solution, the enzyme reaction was carried out at 50 ° C for 5 hours, the enzyme was inactivated at 100 ° C, Engineering / Korea), and the obtained filtrate was concentrated to 40 Brix and used as a fermentation raw material.

Using the YM broth as a basic medium, the combined culture obtained in Example 1-3 was inoculated in the concentrated filtrate, and cultured at 30 DEG C for 24 hours to obtain a fermented product of the perennial plant.

[ Example  1-5] Example  The fermentation broth obtained by 1-3 Powdered

The final culture solution obtained in Example 4 was sterilized at 90 DEG C for 30 minutes, then filtered using a filter press (IJ Engineering / Korea) and pulverized using a spray dryer (Eugene Tech / Korea).

[ Experimental Example  1] 100 years and 100 years Fermented  Compare ingredients

UPLC-QTOF-MS / MS was carried out for each of the fermented products obtained from Examples 1-5 and the index substances of the extracts obtained from Comparative Example 1, respectively.

UPLC was performed by Waters ACQUITY UPLC system and analytical column was ACQUUTY BEH C18 (100 mm x 2.1 mm, 1.8 μm) manufactured by Waters. The mobile phases were (A) tertiary distilled water containing 0.1% formic acid and (B) acetonitrile containing 0.1% formic acid.

As a result (FIG. 1), it was confirmed that the peony root extract and the fermented product of Paekcheon showed completely different peaks, and it was confirmed that the components of the two substances were completely different.

[ Experimental Example  2] 100 years old and 100 years old Fermented  Surface material qualities

The UPLC-QTOF-MS / MS was carried out to qualify the fermented product of Baekryeon obtained in Example 1-5 and the Baekseoncho obtained in Comparative Example 1, respectively.

MS analysis was performed using Q-TOF SNAPT G2 from Waters.

As a result (Fig. 2 and Table 1), it was confirmed that the extracts of Paeoniae sp.

Peak NO. Compound name Retention
time (t _R ). min Formula
[MH] ^- / [M + H] < ⁺ > MS
[MH] ^- / [M + H] < ⁺ >
( m / z) ppm Amount
(mg / 100g) Centipede One Cyanidin-3- (6 " -
feruloylsophoroside) -
5-glucoside 8.32 C ₄₃ H ₄₉ O ₂₄ (+) 949.2589 (+) 1.0 - 2 Cyanidin-3- (6 " - [
coumarylsophoroside
) -5-glucoside 8.52 C ₄₂ H ₄₇ O ₂₃ (+) 919.2482 (+) -1.2 - 3 lsorhamnetin 3-O-
rutinoside 11.37 C ₂₈ H ₃₁ O ₁₆ (-) 623.1612 (-) 0.2 15.5 Fermented water One Caffeic acid 7.16 C ₉ H ₇ O ₄ (-) 179.0344 (-) 0.0 5.4 2 p -Coumaric acid 8.80 C ₉ H ₇ O ₃ (-) 163.0396 (-) 0.6 4.5 3 Ferulic acid 9.82 C ₁₀ H ₉ O ₄ (-) 193.0500 (-) -0.5 10.5 4 Isorhamnetin 3-O-
rutinoside 11.37 C ₂₈ H ₃₁ O ₁₆ (-) 623.1612 (-) 0.2 2.8 5 Isorhamnetin 3-O-
glucoside 11.72 C ₂₁ H ₂₂ O ₁₂ (-) 477,1035 (-) 0.4 1.5

2 (a), most of the compounds identified in the perennial extract are flavonoid glucoside derivatives. On the other hand, the perennial fermented products of the present invention are the graphs of FIG. 2 (b) , It was confirmed that the compound was a phenolic acid-based compound. Therefore, it was confirmed that the present invention analyzed the new indicator substance of the fermented product of Baekryeon.

[Experimental Example 3] Comparison of index substance components of fermented product of Paeoniae sp.

Peak parameter height was set to 5% in each mass spectrometry data (FIG. 1) in order to qualify the index material of the Baengnujang extract obtained in Example 1-5 and the Baengnujang extract obtained in Comparative Example 1, And analyzed by a principle component analysis (PCA) program based on the mass value (m / z) to obtain FIG.

As a result (Fig. 3), fermented products of Paeoniae and Paeoniae were found in clearly distinct regions, respectively, and they were found to contain statistically significant different distributions.

[ Experimental Example  4] Perennial Fermentation product  Depending on the treatment TGF Confirmation of the increase of -β1 concentration

10% FBS, using cultured in 100 units / ml penicillin and 100 ug / ml streptomycin NHDF (normal human dermal fibroblast, Kyung Hee University Skin Biotechnology Center, Republic of Korea) for 37 ℃, 5% CO ₂ conditions in DMEM medium containing Respectively. The NHDF was dispensed into each dish at ⁶ × 10 ⁴ cells / ml and the medium was removed after 24 hours. After that, NHDF was washed with PBS, and 500 μL of phosphate buffered saline was dispensed into each dish and then washed again with PBS. And UVB was irradiated to NHDF under the condition of 144 mJ / cm ² .

Serum-free DMEM medium was dispensed into each dish, and the fermented product of Paecil obtained in Example 1-5 and the Paechelium extract obtained in Comparative Example 1 were treated at a concentration of 1 and 10 μg / mL, respectively. After 72 hours, the supernatant was taken and stored at -20 < 0 >

Cell viability was assessed using MTT assay (Ez-cytox, DOGEN, Korea). Fibroblast cell (Skin Biotechnology Center, Korea) was irradiated with UV light and after 72 hours, the medium was removed, and a plate in which MTT (1 mg / mL) was dispensed for each well was incubated again for 2 hours. The plates containing MTT were measured at 450 nm using a microplate reader (model E09090; Molecular Device, San Francisco, Calif., USA).

The concentration of TGF-β1 was measured by the enzyme-linked immunosorbent assay kit (Duset, R & D system, USA) using the supernatant, and the same experiment was repeated three times. T-test was performed on all experimental data. p < 0.05 (compared to UV untreated group), *; p < 0.05, **; p < 0.01, ***; p < 0.001, and is shown in Fig.

As a result (FIG. 4), it was confirmed that the fermented product of perennial plant increased the concentration of TGF-β1 reduced by UVB, and particularly, it was confirmed that the effect of TGF-β1 was more effective than that of perennial plant.

[ Experimental Example 5 ] Centipedes Fermentation product  Depending on the treatment MMP Confirmation of concentration reduction of -1

(Normal human dermal fibroblast, Kyunghee University Skin Biotechnology Center, Korea) and HaCat-keratinocyte (Kyunghee University Skin Biotechnology Center, Korea) in DMEM medium containing 10% FBS, 100 units / ml penicillin and 100 ug / ml streptomycin. Were cultured at 37 ° C and 5% CO ₂ . The NHDF was dispensed into each dish at ⁶ × 10 ⁴ cells / ml. After 24 hours, the same amount of keratinocyte was added to the dish and cultured for 24 hours. Subsequently, the medium was removed, washed with PBS, and 500 μL of phosphate buffered saline was added to each dish, followed by washing with PBS. UVB was irradiated to the NHDF under the condition of 100 mJ / cm ² . Serum-free DMEM medium was dispensed into each dish, and the fermented product of Paeoniae and the Paeoniae extract obtained by Comparative Example 1 were treated at the concentrations of 0.1, 1, and 5 ㎍ / mL, respectively. After 72 hours, supernatant was taken and stored at -20 ° C and analyzed.

Cell viability was assessed using MTT assay (Ez-cytox, DOGEN, Korea). After incubation at 37 ° C and 5% CO 2 at 37 ° C for 72 hours, the culture medium was removed, and the cells were cultured for 3 days at 37 ° C in a humidified incubator (NHDF) (Normal Human Dermal Fibroblast, Kyunghee University Skin Biotechnology Center, Korea) and HaCat-keratinocyte MTT (1 mg / mL) was dispensed into each well and the plate was incubated again for 2 hours. After 2 hours, the plate containing MTT was again measured at 450 nm using a microplate reader (model E09090; Molecular Device, San Francisco, CA, USA).

The concentration of MMP-1 was measured using an enzyme-linked immunosorbent assay kit (Duset, R & D system, USA) and repeated three times. Data from all experiments were subjected to T-test. p < 0.05 (compared to UV untreated group), *; p < 0.05, **; p < 0.01, ***; p < 0.001, and is shown in Fig.

As shown in FIG. 5, it was confirmed that the concentration of MMP-1 increased by UVB was decreased by treatment with fermented product of Baekjanggi. In particular, it was confirmed that the fermented product of Paekcheon had a better effect than that of Paekcheon in decreasing the concentration of MMP-1.

[ Experimental Example 6 ] Centipedes Fermentation product  Depending on the treatment Percutaneous  Identify changes in moisture content

Six-week old mice (Shizuoka, Japan) were purchased for the experiment, and stabilized for one week. Then, the mice were fed with water and basic diet at 24 ° C and 50% humidity in a light / dark cycle of 12 hours. Animal experiments were conducted in accordance with the Kyung Hee University Animal Resource Experiment Guideline. The certification number was KHUASP (SU) -12-09.

Five experimental groups were used for the experiment.

(a) normal group (N, normal): basic diet supply

(b) Control (C, control): Basic diet + UVB irradiation

(c) N-acetylglucosamine (NAG): UVB irradiation + basic diet + 2.5% NAG,

(d) EO: UVB irradiation + basic diet supply + 2.5%

(e) FO: UVB irradiation + basic diet supply + 2.5% perennial fermentation

All diets were fed as solid diets and were self-administered (Table 2) with sufficient dietary intake per day and UVB was investigated for 9 weeks (Table 3). The weight change and daily average dietary intake of reared mice after 9 weeks are shown in Table 4. Specifically, UVB was examined at 310 nm using Sankyo Denki sunlamps (Kanagawa, Japan). The first week of the experiment was 100 mJ / cm ² once a day and then 200 mJ / cm ² twice a week for 8 weeks. UVB was irradiated on the epidermis of dull mice.

Composition Furtherance (%) Basic diet Cornstarch 35 Maltodextrin 5 Sucrose 20 Casein 20 Soybean oil 5 Lard 5 Mineral Mix 2.5 Vitamin Mix 5 Experimental sample Perennial Seed (EO) 2.5 Fermented water (FO) 2.5 NAG 2.5 Total (%) 100

term Week 1 (daily) Week 2 to 8
(3 times a week) 9 parking
(Once a week) UVB dose
(mJ / cm ² ) 200
(2 minutes 21 seconds) 400
(4 minutes 43 seconds) 200
(2 minutes 21 seconds)

The transdermal water content of each experimental group was measured on the dorsal skin at weekly intervals using a corneometer (MPA-5; Courage Khazaka Electronic GmbH) and all data were averaged and plotted in FIG.

As a result, as shown in FIG. 6, the transdermal water content of all experimental groups was decreased compared with the negative control group without UVB irradiation. However, when the fermented product was administered orally at 4 weeks, NAG The transdermal water content was increased compared to oral administration.

[ Experimental Example 7 ] Centipedes Fermentation product  Confirmed that collagen breakdown decreases with treatment

Each of the test samples of Experimental Example 6 was cut to a size of 7 mm x 7 mm and fixed in 10% formalin buffer. The tissue is then dehydrated using alcohol and placed in paraffin according to the histological examination procedure. To stain the collagen fibers at the site to be stained, the tissue was stained with trichrome stain, and the tissue was observed under a fluorescence microscope at a magnification of 200.

As a result, as shown in FIG. 7, it was confirmed that the control group with UVB irradiation showed more collagen fiber tissue destruction than the normal group without UVB irradiation. However, it was observed that collagen fiber breakage was relatively small in the group (FO) obtained by oral ingestion of the fermented product at 100th percentile, and thus it was confirmed that the fermented product at 100th percentile can inhibit skin aging.

[ Experimental Example 8 ] Centipedes Fermentation product  Depending on processing Pillar green Expression level  Confirm increase

Skin samples of each experimental group of Experimental Example 6 were collected, homogenized in a cell lysis buffer, and proteins were extracted therefrom and western blotting was performed on filaggrin.

Total protein extracts from skin tissues and alpha-tubulin, a Western control sample, were electrophoresed on 10% sodium dodecyl sulfate polyacrylamide gel and transferred to a nitrocellulose membrane (Amersham Pharmacia Biotech, USA). Tissue was obtained from dermis and epidermal tissue using a primary antibody (filaggrin; Santa Cruz Biotechnology) and a secondary antibody (horseradish peroxidase-conjugated secondary antibody; Santa Cruz Biotechnology).

As a result, as shown in FIG. 8, it was confirmed that the expression of filaggrin, which is a precursor of natural moisturizing factor, in the group in which the fermented product of perennial plant was orally ingested (FO was expressed in excellent amounts in each of dermis and epidermis.

Hereinafter, a formulation example of the composition according to the present invention will be described. However, the pharmaceutical composition and the cosmetic composition can be applied to various formulations, which are not intended to limit the present invention but merely to illustrate the present invention.

[Formulation Example 1] Softening lotion (skin lotion)

Compounding ingredient Content (% by weight) Fermented water 0.1 glycerin 3.0 Butylene glycol 2.0 Propylene glycol 2.0 Carboxyvinyl polymer 0.1 Phage-12 nonylphenyl ether 0.2 Polysorbate 80 0.4 ethanol 10.0 Triethanolamine 0.1 Preservative, coloring, fragrance Suitable amount Purified water Balance

[Formulation Example 2] Nutritional lotion (Milk lotion)

Compounding ingredient Content (% by weight) Fermented water 0.5 glycerin 3.0 Butylene glycol 3.0 Propylene glycol 3.0 Carboxyvinyl polymer 0.1 Wax 4.0 Polysorbate 60 1.5 Caprylic / capric triglyceride 5.0 Squalane 5.0 Sorbitacecequioleate 1.5 Liquid paraffin 0.5 Cetearyl alcohol 1.0 Triethanolamine 0.2 Preservative, coloring, fragrance Suitable amount Purified water Balance

[Formulation Example 3] Nourishing cream

Compounding ingredient Content (% by weight) Fermented water 1.0 glycerin 3.0 Butylene glycol 3.0 Liquid paraffin 7.0 Beta Glucan 7.0 Carbomer 0.1 Caprylic / capric triglyceride 3.0 Squalane 5.0 Cetearyl glucoside 1.5 Sorbitan stearate 0.4 Polysorbate 60 1.2 Triethanolamine 0.1 Preservative, coloring, fragrance Suitable amount Purified water Balance

[Formulation Example 4] Massage cream

Compounding ingredient Content (% by weight) Fermented water 1.5 glycerin 8.0 Butylene glycol 4.0 Liquid paraffin 45.0 Beta Glucan 7.0 Carbomer 0.1 Caprylic / capric triglyceride 3.0 Wax 4.0 Cetearyl glucoside 1.5 Sesquioleic acid sorbitan 0.9 Vaseline 3.0 paraffin 1.5 Preservative, coloring, fragrance Suitable amount Purified water Balance

[Formulation Example 5] Pack

Compounding ingredient Content (% by weight) Fermented water 0.1 glycerin 4.0 Polyvinyl alcohol 15.0 Hyaluronic acid extract 5.0 Beta Glucan 7.0 Allantoin 0.1 Nonyl phenyl ether 0.4 Polysorbate 60 1.2 Ethanol preservative 6.0 Good Preservative, coloring, fragrance Suitable amount Purified water Balance

[Formulation Example 6] ointment

Compounding ingredient Content (% by weight) Fermented water 0.1 glycerin 8.0 Butylene glycol 4.0 Liquid 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 Wax 4.0 Preservative, coloring, fragrance Suitable amount Purified water Balance

[Formulation Example 1] Soft capsule

A soft capsule was prepared by mixing 0.0025 g of fermented water at 100 years, 50 mg of red ginseng extract, 2 mg of palm oil, 8 mg of palm kernel oil, 4 mg of yellow pearls, and 6 mg of lecithin and filling them with 400 mg per capsule according to a conventional method.

[Formulation Example 2] Tablets

The granules were formed by mixing 0.0025 g of fermented water, 100 mg of glucose, 50 mg of red ginseng extract, 96 mg of starch and 4 mg of magnesium stearate, and 30% ethanol was added thereto, followed by drying at 60 ° C., Tablets were tableted.

[Formulation Example 3] Granules

The granules were formed by mixing 150 mg of fermented water, 100 mg of glucose, 50 mg of red ginseng extract and 600 mg of starch and 100 mg of 30% ethanol, and dried at 60 ° C. to form granules. The final weight of the contents was 1 g.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (22)

A composition comprising a fermented product,
The perennial fermentation product may be selected from the group consisting of caffeic acid, p-coumaric acid, isorhamnetin 3-O-rutinoside and isocanethin 3-0-glucoside wherein the composition comprises at least one compound selected from the group consisting of isorhamnetin 3-O-glucoside. A composition comprising a fermented product,
The perennial fermentation product may be selected from the group consisting of caffeic acid, p-coumaric acid, isorhamnetin 3-O-rutinoside and isocanethin 3-0-glucoside wherein the composition comprises at least one compound selected from the group consisting of isorhamnetin 3-O-glucoside. delete delete A cosmetic composition for inhibiting skin photoaging by inhibiting the production of collagenolytic enzyme, which comprises fermented water of 100 years old as an active ingredient. 6. The method of claim 5,
The cosmetic composition for inhibiting skin photo-aging by inhibiting the production of collagenase, which is a fermented product of Aureobasidium sp. And Candida sp. The method according to claim 6,
Wherein the Aureobasidium sp. Yeast is Aureobasidium pullulans, which inhibits the production of collagenase. The method according to claim 6,
Wherein the Candida sp. Yeast is Candida utilis. The cosmetic composition for suppressing skin photoaging through inhibition of collagenase production. 6. The method of claim 5,
The perennial fermentation product may be selected from the group consisting of caffeic acid, p-coumaric acid, isorhamnetin 3-O-rutinoside and isocanethin 3-0-glucoside wherein the composition comprises at least one compound selected from the group consisting of isorhamnetin 3-O-glucoside. 6. The method of claim 5,
Wherein the composition comprises 1.0% by weight to 5.0% by weight of the fermented product of perennial plant, based on the total weight of the composition, to inhibit collagenase formation. A pharmaceutical composition for inhibiting skin photoaging by inhibiting the production of collagenolytic enzyme, which comprises fermented water of 100 years old as an active ingredient. 12. The method of claim 11,
Wherein the fermented product is a fermented product of Aureobasidium sp. And Candida sp. Yeast extract of bacillus subtilis extract, which inhibits the production of collagenase. 13. The method of claim 12,
Wherein the Aureobasidium sp. Yeast is Aureobasidium pullulans. The pharmaceutical composition for suppressing skin photoaging through inhibition of collagenase production. 13. The method of claim 12,
The Candida sp. Yeast is Candida utilis, which inhibits the production of collagenase. 12. The method of claim 11,
The perennial fermentation product may be selected from the group consisting of caffeic acid, p-coumaric acid, isorhamnetin 3-O-rutinoside and isocanethin 3-0-glucoside and isorhamnetin 3-O-glucoside. 2. A pharmaceutical composition for suppressing skin photo-aging through inhibition of collagenase degradation, comprising at least one compound selected from the group consisting of isorhamnetin 3-O-glucoside. 12. The method of claim 11,
Wherein the composition comprises 1.0% by weight to 5.0% by weight of the fermented product of perennial plant, based on the total weight of the composition, for inhibiting collagenase degradation. A health food composition for inhibiting skin photoaging by inhibiting the production of collagenolytic enzyme, comprising fermented water of 100 years old as an active ingredient. 18. The method of claim 17,
Wherein the fermented product of the perennial plant is a fermented product of Aureobasidium sp. And Candida sp. Yeast extract of a perennial plant extract, which inhibits the production of collagenolytic enzyme. 19. The method of claim 18,
Wherein the Aureobasidium sp. Yeast is Aureobasidium pullulans, a composition for inhibiting the production of collagen collagenase, and a composition for preventing skin photoaging. 19. The method of claim 18,
The Candida sp. Yeast is Candida utilis, a health food composition for suppressing skin photoaging through inhibition of collagenase production. 18. The method of claim 17,
The perennial fermentation product may be selected from the group consisting of caffeic acid, p-coumaric acid, isorhamnetin 3-O-rutinoside and isocanethin 3-0-glucoside wherein the composition comprises at least one compound selected from the group consisting of isorhamnetin 3-O-glucoside. 18. The method of claim 17,
Wherein the composition comprises 1.0% by weight to 5.0% by weight of the fermented product of perennial plant, based on the total weight of the composition, of the composition for suppressing collagenase formation.

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