KR101244101B1 - Cosmetic composition for improving skin inflammation and including a fermented product of paprika as an effective ingredient - Google Patents

Abstract

The present invention relates to a cosmetic composition for improving skin inflammation containing paprika ferment as an active ingredient, the composition of the present invention inhibits the expression of NF-kB, lipoxygenase and ICAM-1 protein closely related to skin inflammation and erythema By suppressing itching, etc., it has an excellent inflammation improvement effect and can be used safely in the human body.

Description

COSMETIC COMPOSITION FOR IMPROVING SKIN INFLAMMATION AND INCLUDING A FERMENTED PRODUCT OF PAPRIKA AS AN EFFECTIVE INGREDIENT}

The present invention relates to the use of a paprika fermented product (a fermented product), and more particularly to a cosmetic composition for improving skin inflammation comprising the paprika fermented product as an active ingredient.

The skin is a part of the body that is directly exposed to the external environment. Exposure to excessive ultraviolet rays, pollutants or irritants causes skin irritation and inflammatory reactions such as erythema, edema, and itching. The skin trouble caused by the skin irritation stress is not only aesthetic problem, but also the substances produced during the inflammatory reaction incidentally cause pigmentation of the skin and promote the breakdown of the skin elastic fibers to affect the increase of skin wrinkles. Known (Toxicol In Vitro, 18: 231-43 (2004); Irritant dermatitis, 87-111 (1996)).

Until now, antihistamines, vitamin ointments, and corticosteroids are mainly used for the treatment of skin inflammation, but the effects are often temporary and often have severe side effects.

Skin irritation and inflammatory reactions that may occur with the use of cosmetics include irritation such as itching, stinging and burning, erythema, and edema.

In order to alleviate the skin irritation, various methods such as removing skin irritants have been mobilized, but in recent years, functional cosmetics manufactures are increasing, and the functional material itself is often used as a stimulus source. It can be said.

Accordingly, the present inventors studied various natural plants in order to reduce side effects caused by cosmetics and to find substances that are safe for humans and have excellent anti-inflammatory effects. As a result, paprika fermented products are closely related to skin inflammation, NF-kB and lipoxygenase. And by inhibiting the expression of ICAM-1 protein confirmed that it has an excellent effect on the inhibition of dermatitis and completed the present invention based on this.

Accordingly, an object of the present invention relates to a cosmetic composition for improving skin inflammation comprising a paprika fermented product as an active ingredient.

In accordance with the above object, the present invention provides a cosmetic composition for improving skin inflammation comprising a paprika fermented product as an active ingredient.

The cosmetic composition for improving skin inflammation comprising the paprika fermentation product including the paprika fermentation product of the present invention as an active ingredient is excellent in improving inflammation by inhibiting the expression of NF-kB, lipoxygenase and ICAM-1 protein closely related to skin inflammation. It has an effect and can be used safely in the human body.

1 is a graph showing the cytotoxicity test results of the paprika fermentation of the present invention on human skin fibroblasts (Control: untreated cell group, C: paprika control sample treatment group, FSc: paprika fermentation sample treatment group).
2 is a graph showing the degree of inhibition of lipoxygenase by the paprika fermentation product of the present invention (Control: indomethacin treatment group, C: paprika control sample treatment group, FSc: paprika fermentation sample treatment group).
Figure 3 is a graph showing the degree of inhibition of expression of ICAM-1 protein by the paprika fermentation of the present invention (No UV control: UV untreated group, Control: UV treated group, C: Paprika control sample treatment group, FSc: Paprika fermentation Sample treatment group).
4 is a graph showing the degree of inhibition of expression of NF-kB protein by the paprika fermentation product of the present invention (No UV control: no UV treatment group, Control: UV treatment group, C: paprika control sample treatment group, FSc: paprika fermentation) Sample treatment group).

NF-kB (Nuclear factor kappa-light-chain-enhancer of activated B cells) is a transcription factor that modulates various cellular activities such as immune function, acute phase response, and cell cycle regulation. It moves into the nucleus and binds to kB elements of several genes, causing transcription of that gene. NF-kB has an important effect on the development of tumors as well as on immune activity and the inflammatory response system. Stimulation of NF-kB increases the rate of expression of certain genes, leading to an increase in the production of related enzymes and proteins, which abnormally increases the production of inflammatory prostaglandin and other eicosanoids. Increasing NF-kB activity also overactivates the immune system, exacerbating various autoimmune diseases and inflammatory responses. NF-kB stimulation abnormally increases the production and activity of an enzyme called COX (cylooxygenase), which is an important cause of inflammation and cancer. The main activator of NF-kB is free radicals (eg, free radicals derived from UV) (Arch Dermatol Res, 302: 5-17 (2010); The Journal of Clinical Investigation, 107 (1): 3-11 (2001)).

In addition, lipoxygenase is an enzyme involved in the biosynthetic response of various inflammatory and allergic diseases (Advances in Biological Research 2 (3-4): 56-59 (2008); J. Food Sci. Nutr. 4). (3): 163-166 (1999)), ICAM-1 (Intercellular adhesion molecule-1) is a protein produced by vascular endothelial cells and acts to amplify the inflammatory response by acting on the body's movement and concentration of inflammatory cells ( J. Nutr. 134: 1013-1019 (2004); John Wiley & Sons A / S, Experimental Dermatology, 19, e182-e190 (2010)).

The present invention provides a skin cosmetic composition capable of inhibiting the expression of the NF-kB, lipoxygenase and ICAM-1 protein and exerting an effect on the relief of inflammation-related symptoms such as erythema and itching. The composition of the present invention contains paprika fermented product as an active ingredient.

Paprika ( Capsicum annuum var. Angulosum is called Sweet Pepper, Bell Pepper, Pimemto, Paprika, etc., depending on the country. Peppers are classified into Solanaceae family, Capsicum genus, Annuum species, and 6 subspecies: Chili pepper, Capsicum annuum L. var. Acuminatum , Bell or sweet pepper, Capsicum annuum L. var. grossum ), Cone pepper, Capsicum annuum L. var. abbreviatum ), Cherry pepper, Capsicum annuum L. var. cerasiforme )), Red cluster pepper, Capsicuum annum L. var. fasciculatum ), Long pepper, Capsicum annuum L. var. longum ), generally divided into hot pepper and sweet pepper, of which paprika belongs to sweet pepper.

The "paprika" used in the present invention bell peppers (Capsicum including six sub-species, as well as paprika (Capsicum annuum var. Angulosum) of a narrow concept described above Annuum ) is interpreted as a comprehensive concept and is simply abbreviated as Paprika for convenience of explanation.

Paprika fermentation according to the present invention may be prepared by inoculating yeast in paprika raw material or crushed products or extracts of paprika raw material and then cultured together. Specific fermented product manufacturing method may be performed with reference to Korean Patent Application No. 2010-0055218 filed by the present applicant (Miaebu Co., Ltd.). For example, paprika ferment is pulverized paprika raw material for 15 to 30 minutes hydrothermal treatment at 100 to 121 ℃ and incubated for 24 to 72 hours at 30 to 37 ℃, 1 to 2 vvm aeration conditions after inoculation with yeast Can be obtained. At this time, yeast is It can be used by inoculating in an amount of 1 to 2%. Preferably, the paprika fermented product is, after culturing with yeast, extracted for 15 to 30 minutes at 100 to 121 ℃ using a solvent selected from distilled water, ethanol, methanol, hexane and butanol, filtered and concentrated under reduced pressure and then frozen It can be used by drying. Filtration, reduced pressure concentration and lyophilization can be used with methods generally known for natural product extraction.

Usable yeast is Saccaromyces sp .) strain, for example Saccaromyces cerevisiae and Saccharomyces It can be chosen from bayanus . Preferably Saccharomyces cerevisiae MAB Y1 (Accession No .: KCTC 11386BP) deposited by Applicant.

As used herein, “paprika fermented product” is not only a fermentation broth, but also an extract of a fermentation broth, or a concentrate prepared by filtering and / or depressurizing the extract, or a powder prepared by drying (freeze-drying) the concentrate. Can be.

Paprika fermentation in the composition of the present invention may be contained in 0.1 to 2% by weight, preferably 0.5 to 1% by weight based on the total weight of the composition. Meeting the above content range is more effective in alleviating inflammation such as erythema, itching and the like without affecting formulation stability such as phase separation.

The cosmetic composition of the present invention can be prepared in a variety of cosmetic formulations, for example, supple cosmetics, nourishing lotion, nutrition lotion, massage cream, moisturizing cream, nutrition cream, pack, gel, body lotion, body cream, body oil, body It may be formulated as an essence, shampoo, body cleanser, preparation, or skin type cosmetic. The cosmetic composition of the present invention may further include other ingredients conventionally used in the preparation of cosmetics in addition to the paprika fermentation depending on the formulation or purpose of use.

Hereinafter, the present invention will be described in detail with reference to examples. However, this is merely to aid the understanding of the present invention, and the scope of the present invention is not limited to the following examples.

Preparation Example 1 Preparation of Paprika Fermentation Sample

37.5 g of paprika (red eco-friendly agricultural product, Gyeongnam Pearl) was crushed to mix 62.5 ml of distilled water, and then sterilized at 121 ° C. for 15 minutes for culture. Saccharomyces 2% in sterile paprika grind cerevisiae , MAB Y1, KCTC 11386BP) were inoculated and incubated at 30 ° C. for 24 hours.

50% ethanol (150 mL) was mixed with the paprika fermentation broth obtained after the culture, and extracted and sterilized at 121 ° C., filtered, and concentrated under reduced pressure to 1/2 at 80 ° C. The concentrate was lyophilized at −70 ° C. until its weight did not change to obtain 3.1 g of powder (hereinafter referred to as “paprica fermentation sample”).

A paprika extract powder sample obtained by the same process as described above was used as a control sample of the paprika fermentation sample, except that the fermentation process using the yeast was omitted.

< Test Example  1> Cytotoxicity Test ( MTT  analysis)

In order to evaluate the effect of the paprika fermentation sample and the control sample prepared in Preparation Example 1 on the cell survival as follows MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl-tetrazolium bromide) analysis was performed. The MTT assay is a test that takes advantage of the mitochondria's ability to reduce the yellow water-soluble substrate, MTT tetrazolium, to a blue-violet, water-insoluble MTT formazan by dehydrogenase activity. It is well known to reflect the concentration of vigorous cells.

Specifically, human dermal fibroblasts obtained from Clonetics (San Diego, CA, USA) were added Dulbecco's Modified Eagle's Medium (DMEM, Sigma co., St.) containing 10% fetal bovine serum (FBS) and 100 U / ml penicillin. Louis, MO, USA) was incubated at 37 ℃, 5% CO ₂ conditions.

The obtained human dermal fibroblasts were dispensed into each well of a 96 well plate at a density of 10 ⁴ cells / well, and the paprika fermentation sample of Preparation Example 1 at concentrations of 1, 10 and 20 µg / ml during differentiation. FSc) and control sample (C) were treated and then maintained for 4 hours in MTT (1 mg / ml PBS) solution after 48 hours. After dissolving using 0.04 N HCl / isopropanol, the absorbance was measured at 570 nm using a spectrophotometer (ELISA Reader, Bio-Rad, USA).

The results are shown in Figure 1; As shown in FIG. 1 , the paprika control sample was observed to be toxic at 20 μg / ml, but the paprika fermentation sample of the present invention was found to be cytotoxic and safe.

< Test Example  2> Lipoxygenase  Inhibitory efficacy test

Lipoxygenase is an enzyme involved in the biosynthetic response of various inflammatory and allergic diseases. The evaluation of inhibition of lipoxygenase is a commonly used method to determine the extent of skin inflammation reduction in the cosmetics industry (Advances). in Biological Research 2 (3-4): 56-59 (2008)). In this test example, the degree of inhibition of lipoxygenase by the paprika fermentation product of the present invention was evaluated.

Specifically, the paprika fermentation sample (FSc) and paprika control sample (C) prepared in Preparation Example 1 were dissolved in purified water at a concentration of 1, 5, 10, 30 mg / ml. Each sample contains 900 ml borate buffer (0.2M, pH 9), 2 ml linoleic acid (0.6 mM), and 100 ml lipoxygenase (20,000 U / ml) at room temperature for 5 minutes. After the change in absorbance was measured at 234 nm. At this time, the enzyme activity of the purified water was measured to evaluate the degree of inhibition of lipoxygenase in the sample at a relative inhibition rate. As a positive control (Control), indomethacin (indomethacin) effective in anti-inflammatory was used diluted to 20 mg / ㎖.

The results are given in Fig. As shown in Figure 2 , the paprika fermentation sample according to the present invention was shown to be superior to the lipoxygenase inhibitory effect compared to the paprika control sample. In particular, when paprika fermented samples were treated at a concentration of 30 mg / ml, they exhibited high lipoxygenase inhibitory effect comparable to indomethacin.

< Test Example  3> ICAM -1 protein expression inhibition test

Increasing ICAM-1 (Intercellular adhesion molecule-1) in dermal fibroblasts causes infiltration of the skin of cells involved in inflammation such as macrophages and white blood cells. ICAM-1 has been reported to increase the expression of ultraviolet (UV) irradiation (John Wiley & Sons A / S, Experimental Dermatology, 19, e182-e190 (2010)). In this test example, the degree of inhibition of expression of the paprika fermentation sample according to the present invention for the ICAM-1 protein induced by ultraviolet B irradiation was evaluated by Western blot analysis.

Specifically, human dermal fibroblasts obtained from Clonetics (San Diego, CA, USA) were added Dulbecco's Modified Eagle's Medium (DMEM, Sigma co., St.) containing 10% fetal bovine serum (FBS) and 100 U / ml penicillin. Louis, MO, USA) was incubated at 37 ℃, 5% CO ₂ conditions.

Then, the medium was removed and washed with phosphate buffered saline (PBS) to remove the serum components in the medium and irradiated with 100 mJ / cm ² UV-B using a Bio-sun (Vilber Lourmat, Marine, Cedex, France) fluorescent lamp .

Western blot analysis was performed to confirm the expression level of ICAM-1 protein. Specifically, the paprika fermentation sample and the control sample prepared in Preparation Example 1 were treated with UV-treated human skin fibroblasts by concentrations (1, 10, 20 ㎍ / ㎖) and then treated with 150 ml lysis buffer to extract the cell extracts. Prepared and performed 8% SDS-PAGE. To prevent nonspecific binding, 5% skim milk powder was treated, and then the primary antibody of the ICAM-1 protein (monoclonal rabbit anti-human antibody, Santa Cruz Biotech.) Was diluted 1000 times and reacted with stirring, followed by goat anti-rabbit. It was reacted with a secondary antibody of horseradish peroxidase (1: 10000, Jackson ImmunoReasearch Lab.) for 1 hour. ICAM-1 protein in the nitrocellulose membrane was detected by Supersignal West pico chemiluminescence (Pierce Biotech.) And photographed by X-ray film (Konica Co.).

The results are given in Fig. As shown in FIG . 3, in the skin fibroblasts not irradiated with ultraviolet B (No UV control), the expression of ICAM-1 protein was weak, but when the ultraviolet B was irradiated (Control), the expression of ICAM-1 protein was remarkably increased. Increased. When the paprika fermented sample and Paprika control sample of Preparation Example 1 were treated at different concentrations (1, 10, 20 µg / ml) to the fibroblasts irradiated with ultraviolet B, the cell population treated with the paprika fermentation sample of the present invention was obtained. In the case of ICAM-1 protein expression was greatly reduced, especially at concentrations of 10 μg / ㎖ or more showed a similar level of ICAM-1 protein expression level than that without UV treatment.

< Test Example  4> NF - kB  Protein expression inhibition test

NF-kB moves from the cytoplasm to the nucleus and increases in expression when an inflammatory reaction occurs. In this test example, the degree of inhibition of expression of NF-kB by the paprika fermentation product of the present invention was evaluated.

Specifically, the paprika fermentation sample and the control sample prepared in Preparation Example 1 to the skin fibroblasts (300,000 cells / 60 mm dish density) exposed to UV as in Test Example 3 by concentration (1, 10, 20 ㎍ / Ml). 150 ml lysis buffer was added to the cultured cells to prepare cell extracts, followed by 10% SDS-PAGE. After treatment with 5% skim milk to prevent nonspecific binding, mouse primary antibody NF-kB (Cell Signaling Technology Inc., Beverly, MA, USA) was diluted 1000-fold and reacted with stirring, goat anti -rabbit horseradish peroxidase (1: 10000, Jackson ImmunoReasearch Lab., Jackson ImmunoResearch Lab., West Grove, PA, USA) was reacted for 1 hour. Proteins of PPAR gamma and C / EBP alpha in the nitrocellulose membrane were detected by Supersignal West pico chemiluminescence (Pierce Biotech., Rockford, IL, USA) and photographed by X-ray film (Konica Co., Tokyo, Japan).

The results are given in Fig. As shown in FIG . 4 , the NF-kB protein was hardly expressed in the skin fibroblasts (No UV control) which did not irradiate ultraviolet B. However, when the ultraviolet B was irradiated (Control), the expression of the NF-kB protein was remarkably increased. Increased. As a result of treating the paprika fermentation sample according to the present invention to the skin fibroblasts irradiated with ultraviolet B, the expression of NF-kB protein was greatly reduced, especially at a concentration of 10 μg / ml or more, similar to that without UV treatment. NF-kB protein expression levels are shown.

These results show that the composition containing the paprika fermentation product according to the present invention is very effective in alleviating skin inflammation.

<Production Example 1> Nourishing Cream (Milk Skin Lotion)

The nutritional lotion was prepared by the composition of Table 1 according to a conventional method.

ingredient Content (% by weight) Purified water To 100 Paprika Fermentation 0.2 Butylene Glycol 4.0 Hyaluronic acid extract 5.0 Beta Glucan 7.0 Carbomer 0.1 glycerin 8.0 Caprylic Capric Triglycerides 8.0 Squalane 5.0 Cetearyl Glucoside 1.5 Sorbitan stearate 0.4 Cetearyl Alcohol 1.0 Preservative, fragrance, pigment Suitable amount Triethanolamine 0.1

Preparation Example 2 Nutrition Cream

Nutritional cream was prepared in the composition of Table 2 according to a conventional method.

ingredient Content (% by weight) Purified water To 100 Paprika Fermentation 0.5 Butylene glycol 3.0 Liquid paraffin 7.0 Beta Glucan 7.0 Carbomer 0.1 glycerin 3.0 Caprylic Capric Triglycerides 3.0 Squalane 5.0 Cetearyl Glucoside 1.5 Sorbitan stearate 0.4 Polysorbate 60 1.2 Preservative, fragrance, pigment Suitable amount Triethanolamine 0.1

< Manufacturing example  3> pack

According to a conventional method, a pack was prepared with the composition shown in Table 3 below.

ingredient Content (% by weight) Purified water To 100 Paprika Fermentation 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 Preservative, coloring, fragrance Suitable amount

Claims (5)

Paprika extract was inoculated with Saccharomyces cerevisiae MAB Y1 (Accession No .: KCTC 11386BP), fermented at 30 to 37 ° C. for 24 to 72 hours, and then ethanol was added at 100 to 121 ° C. Cosmetic composition for improving skin inflammation comprising paprika fermented product extracted for 15 to 30 minutes as an active ingredient.
The method of claim 1,
The paprika extract is a skin composition for improving skin inflammation, characterized in that the hydrothermal extraction for 15 to 30 minutes at 100 to 121 ℃ by adding distilled water to paprika.
The method of claim 1,
The paprika ferment is a cosmetic composition for improving skin inflammation, characterized by improving inflammation symptoms by inhibiting one or more of NF-kB, lipoxygenase and intercellular adhesion factor-1 (ICAM-1). .
The method of claim 1,
The cosmetic composition is a flexible cosmetics, nourishing cosmetics, nourishing lotion, massage cream, moisturizing cream, nutrition cream, pack, gel, body lotion, body cream, body oil, body essence, shampoo, body cleanser, preparation and skin adhesive type cosmetic Cosmetic composition for improving skin inflammation, characterized in that formed in any one formulation selected from the group consisting of.
The method of claim 1,
The cosmetic composition is a cosmetic composition for improving skin inflammation, characterized in that the paprika fermented product is contained in 0.1 to 2% by weight based on the total weight of the composition.

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