KR102031288B1 - A cosmetic composition comprising cabbage cultiva rfermented extract - Google Patents

Abstract

The present invention provides a cosmetic composition comprising the cabbage fermentation extract excellent in anti-inflammatory activity as an active ingredient.

Description

Cosmetic composition containing cabbage fermented extract as an active ingredient {A COSMETIC COMPOSITION COMPRISING CABBAGE CULTIVA RFERMENTED EXTRACT}

The present invention relates to a cosmetic composition containing the cabbage fermented extract as an active ingredient.

The skin is a membrane that covers the outside of the body and performs various physiological functions that protect the body from environmental factors such as external stimuli, obstacles, and drying. It protects and controls internal organs and other organs. do.

Inflammation is an immune response of the human body in response to a wound or disease, and oxidative stress such as ultraviolet rays, free radicals, and free radicals activates inflammatory factors and causes various diseases and aging of the skin.

Vascular activating polypeptides such as kinin, plasmin, and complement act on vasodilation, contraction and chemotaxis, as well as lymphokines such as interleukin-6 (IL-6). Phosphorus and arachidonic acid are responsible for the inflammatory response. Arachidonic acid is metabolized to inflammatory mediators, prostaglandin and leukotriene, through two pathways, cyclooxygenase or lipooxygenase, to mediate various inflammatory reactions.

Anti-inflammatory drugs act to reduce inflammation, reduce biological response and symptoms in order to eliminate inflammation.

The substances used for anti-inflammatory purposes until now are nonsteroidal flufenamic acid, ibuprofen, benzydamine, indomethacin, and indomethacin, and prednisolone as a steroid system. , Dexamethasone and the like.

In addition, allantoin, azene, hydrocortisone, etc. are known to be effective in anti-inflammatory, but these substances have a limited amount of use as a matter of safety to the skin, or a problem that can not be expected to substantially reduce the effect of the inflammation. There is this.

In addition to the inflammatory response, as the body ages, the secretion of various hormones that regulate metabolism decreases, and the function of immune cells and the activity of cells decreases.

Externally, the ultraviolet rays increase in the sun's rays due to the destruction of the ozone layer, and the excessive physical and chemical irritation and stress caused by the increase of free radicals and active harmful oxygen as environmental pollution becomes more severe are normal functions of the skin. Decreases, promotes the formation of melanin, blemishes, freckles, etc. and skin aging phenomenon will damage the skin.

In order to prevent skin aging and damage caused by these internal and external factors, in order to maintain healthier and more beautiful skin, physiologically active substances obtained from various animals, plants, and microorganisms are added to cosmetics to maintain the unique functions of the skin and Efforts to maintain skin health by activating cells to promote skin metabolism continue, and cosmetics and external preparations for skin are being developed accordingly.

However, the cosmetic product containing the extract derived from the natural method obtained by the conventional method does not fully implement the functionality such as skin whitening effect, there was a problem that the skin improvement activity is not maintained continuously. Therefore, the necessity of developing a natural product-derived cosmetic product with excellent functionality and excellent skin stability and skin improvement is steadily emerging.

The present inventors sought to develop a cosmetic composition of excellent functionality using a natural extract in a situation where the need for the development of a natural product-derived cosmetic product excellent in biosafety and excellent in antioxidant and anti-inflammatory effects.

The present invention is to solve the above problems of the prior art, an object of the present invention to provide an environmentally friendly cosmetic composition excellent in functionality using a natural fermented extract.

According to one aspect of the invention, cabbage ( Brassica) oleracea var. gemmifera ) A cosmetic composition comprising the fermented extract as an active ingredient is provided.

In one embodiment, the fermentation extract may be obtained by inoculating a fermentation strain.

In one embodiment, the fermentation strain may be Marinobacter shengliensis .

In one embodiment, the content of isocyanate may be increased by the fermentation.

In one embodiment, the cosmetic composition may be for anti-inflammatory or anti-stress.

In one embodiment, the cosmetic composition is one selected from the group consisting of skin, lotion, cream, serum, emulsion, essence, powder, foundation, spray, mask pack, seat pack, sleep pack, wash off pack and peel off pack It may be formulated as above.

According to another aspect of the invention, (a) drying and grinding the cabbage by heat sterilization; (b) Marinobacter in the cabbage Marinobacter inoculating and fermenting shengliensis ; (c) drying the fermented cabbage to obtain a solid fermentation product; And (d) immersion extraction by mixing the solid fermentation product and the extraction solvent.

Cosmetic composition according to an aspect of the present invention includes a natural extract as an active ingredient, not only excellent skin improvement effect, but also excellent biosafety.

In addition, since the cosmetic composition of the present invention contains a large amount of the active ingredient by the fermentation process, the skin improvement effect may be remarkably improved.

It is to be understood that the effects of the present invention are not limited to the above effects, and include all effects deduced from the configuration of the invention described in the detailed description or claims of the present invention.

1 is a result of measuring the cytotoxicity by the fermented extract according to an embodiment of the present invention.
Figure 2 is the result of measuring the anti-inflammatory activity of the fermented extract according to an embodiment of the present invention.
Figure 3 is the result of measuring the antistress activity of the fermented extract according to an embodiment of the present invention.
4 and 5 are the results of measuring the sulforaphane content according to the fermentation.

The terminology used herein is to select general terms that are currently widely used as possible in consideration of the functions in the present invention, but may vary according to the intention or precedent of the person skilled in the art, the emergence of new technologies and the like. In addition, in certain cases, there is also a term arbitrarily selected by the applicant, in which case the meaning will be described in detail in the description of the invention. Therefore, the terms used in the present invention should be defined based on the meanings of the terms and the contents throughout the present invention, rather than the names of the simple terms.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art. Terms such as those defined in the commonly used dictionaries should be construed as having meanings consistent with the meanings in the context of the related art, and are not construed in ideal or excessively formal meanings unless expressly defined in this application. Do not.

The numerical range includes the numerical values defined in the range. All maximum numerical limits given throughout this specification include all lower numerical limits as if the lower numerical limits were clearly written. All minimum numerical limits given throughout this specification include all higher numerical limitations as if the higher numerical limit were clearly written. All numerical limitations given throughout this specification will include all better numerical ranges within the broader numerical range, as the narrower numerical limitations are clearly written.

Hereinafter, examples of the present invention will be described in detail, but the present invention is not limited by the following examples.

According to one aspect of the invention, cabbage ( Brassica) oleracea var. gemmifera ) A cosmetic composition comprising the fermented extract as an active ingredient is provided.

The cosmetic composition may include an extract of natural origin as an active ingredient, as well as excellent skin improvement effect and biosafety, and may minimize skin irritation.

The "comprising as an active ingredient" may mean that it contains an effective amount to the extent that can exhibit skin improvement effect activity, such as anti-inflammatory or anti-stress effect.

The "cabbage (Brassica oleracea var. gemmifera ) ”is a perennial or biennial herb of the dicotyledonous poppy family Mustardaceae . It is rich in lysine, an essential amino acid necessary for growth, and contains a large amount of vitamin C, potassium, and calcium. It is also known to clear blood and increase body's resistance to freckles, acne and other skin diseases.

The fermentation extract may be obtained by inoculating a fermentation strain. The "fermentation" refers to a process of decomposing organic matter using an enzyme owned by the fermentation strain. Raw materials that have undergone fermentation through fermentation strains are known to have an effect of at least 2 to 10 times greater than before fermentation.

The fermented metabolite contains various amino acids, organic acids, and antioxidants that are good for the skin to promote skin metabolism to smooth skin texture. In addition, during the fermentation process, the particles become smaller and the toxicity of heavy metals is decomposed, so that the absorption rate is good and the skin troubles or allergic side effects can be alleviated.

The “fermented extract” may be prepared by sterilizing the cabbage and inoculating the fermentation strains, fermenting in predetermined conditions, and then extracting with a specific solvent.

Specifically, the fermentation extract (a) drying and grinding the cabbage by heat sterilization; (b) inoculating the cabbage with Marinobacter and fermenting it; (c) drying the fermented cabbage to obtain a solid fermentation product; And (d) it may be prepared by the step of immersion extraction by mixing the solid fermentation and the extraction solvent.

The “extract” may include all materials irrespective of the extraction method or the type of the component, as long as the active ingredient contained in the extract is separated from the natural ingredient by contacting the solvent and the extract raw material under specific conditions. .

For example, extracting a component dissolved in a solvent from a natural product using water or an organic solvent, and may include both a specific component of the natural product, such as obtained by extracting only a specific component such as oil. The extract may be powdered after drying, or may be obtained by various conventional extraction methods such as hot water extraction and ethanol extraction.

The extract is selected from the group consisting of water, anhydrous or hydrous lower alcohols having 1 to 4 carbon atoms, acetone, petroleum ether, ethyl acetate, butyl acetate, trichloromethane, dichloromethane, chloroform, hexane and 1,3-butylene glycol Which may be extracted with one or more solvents, and specifically with ethanol at a concentration of 60 to 80% (v / v).

The active ingredient contained in the raw material may be different in the extraction ratio according to the polarity of the solvent, the ethanol has excellent selectivity in the extraction of physiologically active substances of the natural raw material to achieve the optimal skin improvement effect by the ethanol extraction Can be.

The extract is washed with water, washed with water, dried and pulverized, and extracted with conventional methods such as reflux circulation extraction, pressure extraction, and ultrasonic extraction for about 1 to 24 hours with a solvent having a volume of 8 to 12 times the weight of the raw material. It can be prepared by filtration. In addition, the extract may be obtained in a powder state by an additional process such as distillation under reduced pressure or freeze drying.

The extract may also include an extract that has undergone conventional purification. For example, the extract may be subjected to various purification methods additionally performed, such as separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity, or affinity). It may comprise a fraction obtained through.

On the other hand, the fermentation strain may be Marinobacter ( Marinobacter ), preferably may be Marinobacter shengliensis ( Marinobacter shengliensis ).

The Marinobacter sengliensis is a Gram-negative bacterium, has a single polar flagella as basophils, aerobic bacilli, and has been isolated from the brine soil of Shengli Oilfield, the second largest heredity in China.

The present inventors promote the hydrolysis of the glucosinolate of the cabbage extract by the fermentation of Marinobacter sengliensis to increase the content of the isocyanate, which is a degradation product, thereby improving the skin such as antistress and anti-inflammatory activity It was confirmed that it can be significantly improved.

The glucosinolate is a group of secondary metabolites whose effects on plant defense and flavor formation against human health, biological and abiotic stresses are well known.

Mustard, horseradish, cabbage, radish, cauliflower, buds, cabbage, kale, broccoli, cabbage, horseradish, and flower buds belonging to the cruciferous vegetable contain various bioactive substances including flavonoids.

The glucosinolate is a secondary metabolite of plants and contains sulfur, nitrogen and the like derived from glucose. About 120 different glucosinolates are known, which can be synthesized from amino acids (tryptophan, methionine, phenylalanine, tyrosine).

The cruciferous plants contain a large amount of glucosinolates and their hydrolysates, indole and isothiocyanate, and a representative component is sulfoaphane.

The sulforaphane isothiocyanate obtained by the glucosinolate is hydrolyzed by the myrosinase during digestion or by the plant myrosinase as the plant cells are destroyed when processing or ingesting the plant. Is a kind of.

The sulforaphane contains ascorbic acid, beta-carotene, β-carotene, rutine, selenium, glutathione, quartose, quercetin, etc. It was first identified as a substance that had activity and had a minimal anti-cancer effect in cruciferous plants by researchers at Johns Hopkins University in 1992 (Zhang Y, et al., Proc. Natl. Acad. Sci. USA 89 (6): 2399403.1992). .

Since the cosmetic composition contains a large amount of isothiocyanate, which is a degradation product of the glucosinolate, the anti-inflammatory or antistress activity is excellent.

Said "anti-inflammatory" means the inhibition or amelioration of inflammation. The "inflammation" is a mechanism for repairing and repairing the damaged area when an invasion that causes any organic change, such as physical action, chemicals, or bacterial infection, is applied to the living body or tissue. Vascularly active substances such as serotonin, bradykinin, prostaglandins, hydroxyeicosatetraenoic acid (HETE), and leukotriene are released and inflammation can be induced by increased vascular permeability.

The cosmetic composition may be formulated into one or more selected from the group consisting of skins, lotions, creams, serums, emulsions, essences, powders, foundations, sprays, mask packs, seat packs, sleep packs, wash off packs and peel off packs. have.

The cosmetic composition may be formulated by conventional methods. Reference may be made to the formulations disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA, for formulation of external skin preparations, and in International Cosmetic Ingredient Dictionary, 6th ed (The cosmetic, Toiletry and Fragrance Association). , Inc., Washington, 1995).

Specifically, the cosmetic composition may be prepared in a general emulsion formulation and solubilized formulation. For example, lotion, such as a flexible lotion or a nourishing lotion; Latex, such as facial lotion and body lotion; Creams such as nutrition creams, moisture creams and eye creams; essence; Cosmetic ointments; spray; Gels; pack; Sunscreen; Makeup base; Foundations such as liquid type, solid type or spray type; powder; Makeup removers such as cleansing cream, cleansing lotion and cleansing oil; Or it may be formulated as a cleaning agent such as cleansing foam, soap, body wash, but is not limited thereto. In addition, the topical skin preparation may be formulated as an ointment, patch, gel, cream or spray, but is not limited thereto.

The cosmetic composition may be appropriately blended with other ingredients within the range of not impairing the object according to the present invention, depending on the kind or purpose of use of the formulation in addition to the essential components in each formulation.

The cosmetic composition may generally include an acceptable carrier, for example, oil, water, surfactants, moisturizers, lower alcohols, thickeners, chelating agents, pigments, preservatives, fragrances, etc. may be appropriately blended, but is not limited thereto. no.

The acceptable carrier may vary depending on the formulation. For example, when formulated into ointments, pastes, creams or gels, as carrier ingredients, animal oils, vegetable oils, waxes, paraffins, starches, trachants, cellulose derivatives, polyethylene glycols, silicones, bentonites, silica, talc, zinc oxide or theirs Mixtures can be used

When the cosmetic composition is formulated as a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder or mixtures thereof may be used as the carrier component, and in the case of a spray, chlorofluorohydro It may further comprise a propellant such as carbon, propane, butane or dimethyl ether.

When the cosmetic composition is formulated into a solution or emulsion, a solvent, solubilizer, or emulsion agent may be used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl benzoate, propylene glycol, 1,3-butylglycol oil may be used, and in particular, cottonseed oil, peanut oil, corn seed oil, olive oil, castor oil and sesame oil, glycerol aliphatic esters, fatty acid esters of polyethylene glycol or sorbitan may be used.

When the cosmetic composition is formulated as a suspension, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Microcrystalline cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used.

When the cosmetic composition is formulated with a soap, as a carrier component, alkali metal salts of fatty acids, fatty acid hemiester salts, fatty acid protein hydrolyzates, isethionates, lanolin derivatives, aliphatic alcohols, vegetable oils, glycerol, sugars and the like Can be used.

The cosmetic composition is a fatty material, organic solvent, solubilizer, thickening agent, gelling agent, softener, antioxidant, suspending agent, stabilizer, foaming agent, fragrance which are commonly used in the industry according to the quality or function of the final product. Commonly used in surfactants, water, ionic or nonionic emulsifiers, fillers, metal ion rod blockers, chelating agents, preservatives, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, cosmetics It may additionally contain adjuvants commonly used in the cosmetic or dermatological arts, such as any other ingredient.

However, the adjuvant and its mixing ratio may be appropriately selected so as not to affect the desirable properties of the cosmetic composition according to the present invention.

Through the following examples, the present invention will be described in more detail, but the present invention is not limited by the following examples.

Example: Preparation of Cabbage Fermented Extract

The cabbage was washed with water, dried at room temperature and ground to give 100 g of a ground product.

The cabbage grounds were autoclaved at 121 ° C. for 20 minutes and then inoculated with Marinobacter shengliensis mycelium.

Cabbage grounds inoculated with the fermentation strains were incubated at 30 ° C. for 5 days, maintaining a humidity of 80 to 90%.

The fermented cabbage was extracted twice at 80 ° C. for 2 hours with a 10-fold volume of 70% (v / v) ethanol as a solvent.

The extract was filtered through a 250 mesh and 0.5 μm filter, concentrated under reduced pressure at 40 ° C., and lyophilized to obtain a solid of each of the willow stem fermentation extract.

Comparative Example 1: Preparation of Cabbage Extract

The cabbage was washed with water, dried at room temperature and ground to give 100 g of a ground product. To 100 g of the pulverized product, 10 times volume of 70% (v / v) ethanol was extracted twice at 80 ° C. for 2 hours.

The extract was filtered through a 250 mesh and 3 ㎛ filter, concentrated under reduced pressure at 40 ℃ and lyophilized to obtain a solid of the extract.

Comparative Example 2: Bacillus fermented extract preparation

Fermentation extracts were prepared in the same manner as in Example, except that only fermentation strains were Bacillus subtilis .

Comparative Example 3 Preparation of Lactobacillus Fermented Extract

A fermented extract was prepared in the same manner as in Example, but only the fermented strain was changed to Lactobacillus casei .

Experimental Example 1: Evaluation of Skin Safety

In order to verify the skin safety of the samples of Examples and Comparative Examples, MTT assay experiments on fibroblasts (HDF), melanocytes (B16F10), keratinocytes (HaCaT) was performed.

Samples of Examples and Comparative Examples were suspended in purified water at concentrations of 0, 1, 5, 10, 20, 50 and 100 mg / L, respectively, and cell viability was measured.

Cytotoxicity was performed by modifying Mosmann's method of measuring cell viability using MTT {3- (4,5-dimethylthiazol-2-yl) -2-5-diphenyltetrazolium bromide} reagent.

Fibroblasts (HDF), melanocytes (B16F10), and keratinocytes (HaCaT) were each dispensed in 96-well plates at a concentration of 1 × 10 ⁴ cells / well and incubated at 37 ° C., 5% CO ₂ for 48 hours. It was.

After removing the culture medium and incubating the sample for 48 hours in a concentration-treated medium, the medium was removed and washed repeatedly with PBS (Phosphate buffered saline).

MTT was dissolved in PBS at 5 mg / mL, 50 μL was added, and incubated for 48 hours at 37 ° C. and 5% CO ₂ . 100 L of DMSO (dimethyl sulfoxide) was added per well, stirred for 10 minutes, and the absorbance was measured at 540 nm.

As a result, no cytotoxicity was confirmed at all concentrations of each sample (FIG. 1). The results suggest that the sample is not only harmless to the human body but also has excellent human safety.

Experimental Example 2 Evaluation of NO Production Inhibition Effect

In order to confirm the anti-inflammatory effects of the Examples and Comparative Examples, nitric oxide (NO) formation inhibitory experiments were performed by the GRIESS method using RAW264.7 cell line (ATCC number: CRL-2278).

Specifically, RAW264.7 cells, which are macrophages of mice, were passaged several times, placed in 24-well plates each containing 3 × 10 ⁵ cells in one well, and then cultured for 24 hours.

It was replaced with the cell medium containing the samples of the Examples and Comparative Examples diluted by concentration. The positive control group was incubated for 30 minutes by treating 2-amino-4-picoline, a NO production inhibitor, with 5 μM, and incubating with 1 μg of LPS (Lipopolysaccharide) for 24 hours.

100 μL of the supernatant was taken, transferred to a 96-well plate, 100 μL of GRIESS solution was added thereto, and reacted at room temperature for 10 minutes. The absorbance at 540 nm was measured to determine the effect of inhibiting NO production of the extract (FIG. 2).

The experiment was repeated three times, the inhibition of NO production can be evaluated as excellent anti-inflammatory effect.

In the case of inoculation of Marinobacter sengliensis, the NO production rate was significantly lower than that of Comparative Examples 1 to 3, which suggests that the anti-inflammatory effect may be significantly increased by Marinobacter sengliene fermentation.

Experimental Example 3 Evaluation of Stress Inhibitory Effect

The inhibitory effect of COX-2 (cyclooxygenase-2) on the expression level with inflammatory response was evaluated.

The keratinocytes (HaCaT) were incubated for 24 hours in a 12-well plate, followed by transfection of the luciferase vector into which the COX-2 gene was inserted.

COX-2 production was induced by treating 10 ㎍ / mL benzopyrene (B [a] P) after treating the cultured cells with different concentrations of Examples and Comparative Examples.

The degree of COX-2 gene expression inhibited by the test substance was measured using the luciferase fluorescence expression level (FIG. 3).

Referring to FIG. 3, the expression of the increased COX-2 gene by benzopyrene was effectively inhibited by the sample of the example and concentration-dependently reduced the expression of the COX-2 gene in keratinocytes.

Experimental Example 4: Evaluation of isothiocyanate content through HPLC quantitative analysis

DL-sulforaphane, allyl isothiocyanate (AITC), benzyl isothiocyanate (BITC), phenethyl HPLC analysis was performed on isothiocyanate (PEITC) standards and analytical samples for the fermented extracts were prepared and compared.

Standard solutions and assay samples were dissolved in 70% ACN solution and filtered through a syringe filter. HPLC analysis was carried out by operating the mobile phase conditions at a flow rate of 0.5 mL / min, a sample temperature and a column temperature of 25 ℃ of methanol: purified water ratio of 85: 15, and the absorbance of 250nm wavelength was measured.

When the standard solution was analyzed under these conditions, the retention time of each isothiocyanate was found to be around 5.5 minutes for AITC, 7 minutes for BITC, 7.2 minutes for PEITC, and 3.8 minutes for DL-sulforaphane.

The change in the content of DL-sulforaphane was compared with before and after fermentation.

4 and 5, the sulforaphane content was significantly increased in the sample of the Example fermented with Marinobacter sengliensis. In addition, the sulforaphane change tendency correlated with the evaluation results of Experimental Examples 2 and 3.

The foregoing description of the present invention is intended for illustration, and it will be understood by those skilled in the art that the present invention may be easily modified in other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are exemplary in all respects and not restrictive. For example, each component described as a single type may be implemented in a distributed manner, and similarly, components described as distributed may be implemented in a combined form.

The scope of the invention is indicated by the following claims, and it should be construed that all changes or modifications derived from the meaning and scope of the claims and their equivalents are included in the scope of the invention.

Claims (7)

Cosmetic composition comprising a cabbage ( Brassica oleracea var. Gemmifera ) fermentation extract obtained by inoculating Marinobacter shengliensis as an active ingredient. delete delete The method of claim 1,
Cosmetic composition, wherein the content of isocyanate is increased by the fermentation. The method according to any one of claims 1 to 4,
The cosmetic composition is for anti-oxidation or inflammation relief, cosmetic composition. The method according to any one of claims 1 to 4,
The cosmetic composition is formulated with one or more selected from the group consisting of skin, lotion, cream, serum, emulsion, essence, powder, foundation, spray, mask pack, sheet pack, sleep pack, wash off pack and peel off pack Composition. (a) drying and grinding the cabbage to heat sterilization;
(b) Marinobacter in the cabbage Marinobacter inoculating and fermenting shengliensis ;
(c) drying the fermented cabbage to obtain a solid fermentation product; And
(D) a step of immersion extraction by mixing the solid fermentation and the extraction solvent; Method of producing a cabbage fermentation extract comprising a.

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