KR102471026B1 - Cosmetic composition containing Cyperus spp. extract for antiinflammation or skin whitening - Google Patents

Abstract

The present invention relates to a composition containing a plant extract of the genus Cyperus , wherein the plant of the genus Cyperus is Cyperus nipponicus , Cyperus microiria , Cyperus difformis , At least one species is selected from the group consisting of Cyperus orthostachyus and Cyperus flavidus , among which the leaves and stems inhibit the activity of inflammation-related substances and tyrosinase to improve skin conditions Multifunctional cosmetic composition readily available as

Description

Cosmetic composition containing Cyperus genus plant extract for anti-inflammatory or skin whitening {Cosmetic composition containing Cyperus spp. extract for antiinflammation or skin whitening}

The present invention relates to a cosmetic composition for anti-inflammatory or skin whitening containing a Cyperus spp. plant extract. The plant of the Cyperus genus is a blue beak ( Cyperus nipponicus ), a golden beetle ( Cyperus microiria ), an egg beetle ( Cyperus difformis ), a beetle beetle ( Cyperus orthostachyus ) and a dreungbang beetle ( Cyperus flavidus ) One or more species may be selected from the group consisting of have.

Hyperpigmentation of the skin is caused by various factors such as hormonal abnormalities in the body after an inflammatory reaction of the skin, genetic diseases, and ultraviolet irradiation, and the main factor is due to abnormal melanin synthesis and distribution. Melanin is produced by converting tyrosine into dopaquinone by the action of tyrosinase in melanocytes, and then converting the dopaquinone into melanin by the action of enzymes and spontaneous oxidation reactions.

Methods for inhibiting melanin production include the following methods. The first method is to remove the main cause of melanin production by blocking ultraviolet rays, and a whitening reaction can be induced by preparing a cosmetic containing a light scattering agent or a light blocking agent. . The second method is a method of inhibiting melanin production by inhibiting the synthesis of core carbohydrates required for tyrosinase to be active, and a compound such as glucosamine may be used. A third method is a method of interfering with cell division of the melanocytes by using a substance having specific toxicity to melanocytes, and a compound such as hydroquinone may be used. A fourth method is a method of interfering with the function of tyrosinase, an enzyme involved in melanin production, and a compound such as kojic acid or arbutin may be used. As another method, there is also a method of reducing the produced melanin. When using compounds such as hydroquinone, kojic acid, or arbutin in the above method, compounds derived from natural products may be used, but synthetic compounds may also be used. Representatively used synthetic compounds include 4-n-butylresorcinol, quercetin, kaempferol, etc., in addition to hydroquinone and kojic acid or arbutin mentioned above. Although the synthetic compounds have good whitening effects, their use has been limited due to product safety problems caused by skin irritation. In particular, synthetic 4-n-butylresorcinol is easily browned due to poor stability during product production, and skin irritation may occur when used in high concentrations. Therefore, in order to solve these disadvantages, a method of liposomizing 4-n-butylresorcinol using gamma-linolenic acid has been disclosed (Korean Registered Patent No. 10-0751883), but it was impossible to completely eliminate irritation. .

The inflammatory response is one of the body's defense mechanisms against causes such as physical stimulation, chemical stimulation, and bacterial infection, and is one of the mechanisms for removing external substances and restoring and regenerating damaged areas. Once stimuli associated with the inflammatory response are applied, vasoactive substances such as histamine, serotonine, bradykinin, prostaglandin, HETE (hydroxyeicosatetraenoic acid), and leukotriene are released locally. Inflammation is induced as vascular permeability increases.

Monocytes are activated by bacterial infection and change into macrophages that perform phagocytosis. Macrophages induce inflammation by secreting NO (nitric oxide; nitrogen monoxide), prostaglandin E2, and pro-inflammatory cytokines. It acts as an important regulatory cell in all immune responses, and macrophages must go through an activation process in order to function like this. LPS (lipopolysaccharide), which is one of the cell wall constituents of Gram-negative bacteria and is well known as an endotoxin, is the most well-known extrinsic factor involved in the activation of macrophages. In particular, pro-inflammatory cytokines (pro-inflammatory cytokines such as TNF-α (tumor necrosis factor-alpha), IL-6 (interleukin-6), and IL-1β (interleukin-1 beta)) are inhibited in monocytes or macrophages such as RAW 264.7 cells. -Inflammatory cytokine) is secreted to increase LPS (lipopolysaccharide) at the inflammatory site. When LPS stimulates macrophages, an enzyme called iNOS (inducible nitric oxide synthase) converts L-arginine into L-citrulline, and NO is produced, and macrophages release NO. generate

In mammals, NO is synthesized by three types of NO synthase (NOS, nitric oxide synthease): neuronal NOS (nNOS), endothelial NOS (eNOS), and inducible NOS (iNOS). Among these, NO produced by nNOS and eNOS is produced for normal biological functions, and its concentration in tissues is maintained at a low level at a constant level. However, the amount of NO generated by iNOS is excessive and exhibits harmful effects to the living body such as pathological vasodilation, cytotoxicity, and tissue damage. In addition, prostaglandin E2 (PGE2), an inflammatory factor, stimulates phospholipids, a component of the cell membrane, by LPS, and arachidonic acid, which is released by an enzyme called phospholipase A2, catalyzes an enzyme called COXs (cyclooxygenase). is formed and induces an inflammatory response. COX is classified into COX-1 and COX-2. COX-1 acts on normal biological functions such as platelet formation, gastric wall protection, and maintenance of kidney function in the body, and COX-2 synthesizes PHE2, an inflammatory mediator. PHE2 is known to be deeply involved in cancer development by promoting inflammatory responses (pain, fever, etc.), immune responses, and angiogenesis. In most cases, iNOS inhibitory drugs have a mechanism of action that is linked to COXs inhibitory substances. Therefore, in many cases, iNOS inhibitors, which are relatively easy to search for, are discovered and developed as anti-inflammatory materials.

In addition, non-steroidal anti-inflammatory drugs (NSAIDs) including selective COX-2 inhibitors exhibit anti-inflammatory, antipyretic and analgesic effects. However, long-term use of these drugs can sometimes cause serious side effects. For example, secondary anemia caused by bleeding from peptic ulcer in the gastrointestinal system, inhibition of platelet function, inhibition of labor induction, side effects on the kidneys, liver damage, and hypersensitivity reactions.

In order to overcome the side effects of these drugs, preference is given to products derived from nature, such as plants, as safer materials, and in this regard, studies are being actively conducted to find and utilize anti-inflammatory materials from natural products.

Cyperus flavidus grows in damp places on plains. The stems are gathered in several patterns, stand upright, and are thin and hard. It is 20-40 cm in height. The leaf is 1-2mm wide, dark green, shorter than the flower stem, and the leaf sheath is dark coppery brown. Flowers bloom in August-October, and bracts are 2-4, and the lower one is longer than the flower ear, and the flower ear is umbrella-shaped or head-shaped. The flower ears are 3-12 cm in length and width, and the long branches reach 8 cm in length. At the end, 5-10 spikelets hang in the shape of a palm, and the spikelets at the bottom are line-shaped lanceolate. The scale pieces are egg-shaped and oval, and the end is dull. The fruit is an achene, flat, shaped like an upside-down egg, and brown. The stigma is two. Distributed in Korea and Japan.

The blue cockroach ( Cyperus nipponicus ) is 10 to 30 cm high, and the leaves come from the root and are narrow and linear. Flowers bloom in raceme in August-September, and fruits bear achene. It grows in wetlands and is distributed in Korea, Japan, and China.

Golden copper ( Cyperus microiria ) is 20 to 30 cm tall, and the leaves are alternate and threadlike. In July-August, yellowish brown spikelets bloom as an umbel at the end of the stem, and fruits bear achene. It grows in rice fields and is distributed in Korea, Japan, and China.

Cyperus difformis is 25 to 60 cm tall and has linear leaves. In August-October, the head-shaped flowers at the end of the main stem bloom densely at the end of the stem in an umbel, and the fruit is an achene. It grows in rice fields or wetlands, and is widely distributed in temperate and temperate regions, and is distributed throughout Korea.

Cyperus orthostachyus is 20 to 60 cm tall, and leaves are often formed from roots and are linear. From August to October, 8 to 20 flowers bloom at the end of the peduncle from between 3 to 5 bracts attached to the end of the stem. The fruit bears about 3mm of fruit. It grows on rice field banks or wetlands, and is distributed in Korea, Japan, and China.

Accordingly, the inventors of the present invention completed the present invention by confirming that the five species of Cyperus plant extracts had excellent anti-inflammatory function and skin whitening effect while studying the physiological activities of various Cyperus plant extracts.

Republic of Korea Patent Registration No. 10-1908221 (Title of Invention: Anti-Obesity Composition Containing Golden Dongsani Extract, Applicant: Dong-Eui University Industry-University Cooperation Foundation) Republic of Korea Patent Publication No. 10-2015-0026579 (Title of Invention: Pharmaceutical composition and health functional food for the prevention and treatment of allergic or non-allergic skin diseases containing herbal extracts as active ingredients, Applicant: YD Life Science Co., Ltd. ) Republic of Korea Patent Registration No. 10-0751883 (Title of invention: Whitening cosmetic composition containing gamma-linolenic acid-4-n-butylresorcinol water-soluble nanoliposome, Applicant: Wellskin Co., Ltd.) Republic of Korea Patent Publication No. 10-2013-0062112 (Title of Invention: External skin composition for prevention and treatment of inflammatory diseases containing hyangbuja extract as an active ingredient, Applicant: Korea Institute of Oriental Medicine Industry Promotion Foundation) Republic of Korea Patent Publication No. 10-2017-0137465 (Title of Invention: Composition for Skin Improvement Containing Complex Herbal Medicine Extract, Applicant: LG Household & Health Care Co., Ltd.)

An object of the present invention is to provide a cosmetic composition for anti-inflammatory or skin whitening containing a Cyperus spp. plant extract. The plant of the Cyperus genus is a blue beak ( Cyperus nipponicus ), a golden beetle ( Cyperus microiria ), an egg beetle ( Cyperus difformis ), a beetle beetle ( Cyperus orthostachyus ) and a dreungbang beetle ( Cyperus flavidus ) One or more species may be selected from the group consisting of have.

The present invention relates to a cosmetic composition for anti-inflammatory or skin whitening containing a Cyperus spp. plant extract. The plant of the Cyperus genus is a blue beak ( Cyperus nipponicus ), a golden beetle ( Cyperus microiria ), an egg beetle ( Cyperus difformis ), a beetle beetle ( Cyperus orthostachyus ) and a dreungbang beetle ( Cyperus flavidus ) One or more species may be selected from the group consisting of have.

The extract is characterized by inhibiting the production of nitric oxide (NO), interleukin-6 (IL-6) or tumor necrosis factor-α (TNF-α).

The extract has tyrosinase inhibitory activity.

The extract is characterized in that it is extracted using water, C1-C4 alcohol or a mixed solution thereof as a solvent. The C1-C4 alcohol may be selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol and isobutanol.

The cosmetic composition is skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrient lotion, massage cream, nutrient cream, moisture cream, hand cream, foundation, essence, nutrient essence, pack, soap , It may have any one formulation selected from the group consisting of cleansing foam, cleansing lotion, cleansing cream, body lotion and body cleanser.

In addition, the present invention may relate to a health functional food for anti-inflammatory or skin whitening containing a Cyperus spp. plant extract.

Hereinafter, the present invention will be described in detail.

The plant extract of the genus Cyperus may be extracted using an extraction solvent from leaves, stems, rhizomes, flowers, fruits, or mixtures thereof of the collected plants. Preferably, a mixture of leaves and stems in a weight ratio of 1:0.5 to 1:2 may be used.

The Siperus genus plant extract can be prepared by using water, C1-C4 alcohol, or a mixed solution thereof as a solvent. As the water, any water without impurities such as distilled water and ionized water (DI water) may be used. The C1-C4 alcohol may be selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol and isobutanol. At this time, the plant extract of the genus Cypherus is preferably a C1-C4 alcohol extract or an aqueous solution extract of C1-C4 alcohol rather than a water extract, and an aqueous solution extract of C1-C4 alcohol has the best anti-inflammatory or skin whitening activity. Suitable as an aqueous solution of C1-C4 alcohol is a 50 to 90% (v/v) aqueous ethanol solution, and most suitable is a 60 to 70% (v/v) aqueous ethanol solution.

In preparing the Siperus genus plant extract, extraction may be performed using a solvent of 5 to 40 times, preferably 10 to 20 times the weight of the Siperus plant dry weight. If the solvent is too less than the above range, when preparing the extract by applying heat, extraction may not be performed well due to evaporation of the solvent, and if the solvent is too large, there is a problem in that the extraction time is too long.

In addition, the extraction process may be carried out while stirring at 10 to 40 ° C. for 24 to 48 hours, and extraction may be repeated 1 to 4 times to obtain a liquid extract.

The liquid extract may be filtered out using sterile gauze or filter paper. The filtrate through this filtration process can be concentrated under reduced pressure at 10 to 40 ° C using a rotary vacuum concentrator, and an extract can be obtained after removing the remaining solvent by lyophilization.

The Siperus genus plant extract may be included in the cosmetic composition in an amount of 0.01 to 20% by weight. If the Siperus genus plant extract is included in the cosmetic in an amount of less than 0.01% by weight, the anti-inflammatory or skin whitening effect may be insignificant, and if it exceeds 20% by weight, formulation may be difficult. In addition, if it exceeds 10% by weight, the degree of improvement in anti-inflammatory or skin whitening functionality may be insignificant, so that the Siperus genus plant extract may be preferably included in the cosmetic composition in an amount of 0.01 to 10% by weight. At this time, the weight of the plant extract of the genus Cyperus is based on the dried powder form of the plant extract of the genus Cyperus.

In addition, the present invention may provide a cosmetic formulation containing the cosmetic composition as a raw material or an active ingredient. The formulations are external skin ointment, essence, whitening cream, lotion, emulsion, pack, general lotion, skin milk, skin, cream, serum, beauty soap, softening lotion, medicated lotion, body cleanser, cleansing oil, cleansing cream, cleansing tissue And cleansing water can be provided.

In the cosmetic formulation of the present invention, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, lactose, silica, aluminum hydroxide, Calcium silicate, polyamide powder, chlorofluorohydrocarbon, propane/butane or dimethyl ether, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic esters, polyethylene glycol or fatty acid esters of sorbitan, ethoxylated isostearyl alcohols, suspending agents such as polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonites, Agar or tracanth, aliphatic alcohol sulfates, aliphatic alcohol ether sulfates, sulfosuccinic acid monoesters, isethionates, imidazolinium derivatives, methyl taurates, sarcosinates, fatty acid amide ether sulfates, alkylamidobetaines, fatty acid glycerides , fatty acid diethanolamide, lanolin derivatives, or ethoxylated glycerol fatty acid esters may be used.

The present invention may be provided as an anti-inflammatory pharmaceutical composition containing a plant extract of the genus Cyperus. In addition, the extract can also be provided as a pharmaceutical composition for preventing or improving inflammatory diseases. The inflammatory disease consists of inflammatory bowel disease, inflammatory collagen vascular disease, glomerulonephritis, sarcoidosis, atopic dermatitis, retinitis, gastritis, hepatitis, enteritis, arthritis, tonsillitis, sore throat, bronchitis, pneumonia, pancreatitis, cystitis, neuritis, sepsis and nephritis. can be selected from the group.

The pharmaceutical composition may be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories and sterile injection solutions according to conventional methods, respectively. Carriers, excipients, and diluents that may be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, and cellulose. , methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include at least one excipient for the extract of the present invention, for example, starch, calcium carbonate, sucrose or lactose, It is prepared by mixing gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, solutions for oral use, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, aromatics, and preservatives may be included. . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried formulations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspensions. As a base for the suppository, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerogeratin and the like may be used.

The dosage of the pharmaceutical composition of the present invention will vary depending on the age, sex, and weight of the subject to be treated, the specific disease or pathological condition to be treated, the severity of the disease or pathological condition, the route of administration, and the judgment of the prescriber. Determination of dosage based on these factors is within the level of those skilled in the art, and generally dosages range from 0.01 mg/kg/day to approximately 2000 mg/kg/day. A preferred dosage is 1 mg/kg/day to 500 mg/kg/day. Administration may be administered once a day, or may be administered in several divided doses. The dosage is not intended to limit the scope of the present invention in any way.

The pharmaceutical composition of the present invention can be administered to mammals such as rats, livestock, and humans through various routes. All modes of administration are contemplated, eg oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine intrathecal or intracerebrovascular injection. Since the extract of the present invention has little toxicity and side effects, it is a drug that can be safely used even when taken for a long period of time for preventive purposes.

In addition, the present invention provides a health functional food for anti-inflammatory or skin whitening containing a plant extract of the genus Cyperus. The extract can also be provided as a health functional food for preventing or improving inflammatory diseases.

The Siperus genus plant extract may be added to the health functional food of the present invention in an amount of 0.01 to 100% by weight. The health functional food of the present invention includes forms such as tablets, capsules, pills or liquids, and foods to which the extract of the present invention can be added include, for example, various drinks, meat, sausages, bread, candy, Snacks, noodles, ice cream, dairy products, soups, ionic beverages, soft drinks, alcoholic beverages, chewing gum, tea, and vitamin complexes.

The present invention relates to a composition containing a plant extract of the genus Cyperus , wherein the plant of the genus Cyperus is Cyperus nipponicus , Cyperus microiria , Cyperus difformis , At least one species is selected from the group consisting of Cyperus orthostachyus and Cyperus flavidus , among which the leaves and stems inhibit the activity of inflammation-related substances and tyrosinase to improve skin conditions Multifunctional cosmetic composition readily available as

1 is a graph showing the results of CCK-8 experiments confirming the cytotoxicity of plant extracts of the genus Cypherus.
Figure 2 is a graph showing the experimental results confirming the NO scavenging ability of the plant extract of the genus Cypherus.
3 is a graph showing the IL-6 production inhibitory effect of plant extracts of the genus Cyperus.
Figure 4 is a graph showing the production inhibitory effect of TNF-α of plant extracts of the genus Cypherus.

Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, it is provided to sufficiently convey the spirit of the invention to those skilled in the art, so that the disclosure herein will be thorough and complete.

Preparation Examples 1 to 7. Preparation of plant extracts of the genus Siperus

For the preparation of plant extracts of the genus Cyperus of the present invention, 5 species of plants of the genus Cyperus, Cyperus nipponicus , golden brassicas ( Cyperus microiria ), egg chambers ( Cyperus difformis ), cyperus orthostachyus ( A mixture of equal amounts of leaves and stems of Cyperus flavidus ) was washed with tertiary distilled water and pulverized using a pea. Extraction was performed while stirring at 25 ° C. for 24 hours by adding 10 times the weight of a 70% (v / v) ethanol aqueous solution to a mixture of the leaves and stems of the pulverized Cyperus genus in equal amounts. Thereafter, the filtrate was first filtered using sterile gauze, and then the filtrate was filtered again using filter paper (Whatman, No. 2: Whatman Ltd., Maidstone, Kent, UK), and the pulverized sample was filtered using a rotary vacuum concentrator. After concentration under reduced pressure, the remaining solvent was removed with a lyophilizer to obtain a plant extract of the genus Siferus. In addition, as shown in Table 1 below, extracts of Cyperus pacificus or Cyperus rotundus were prepared using samples from the same region.

Condition sample Preparation Example 1 blue cockroach Preparation Example 2 Golden Dongsani Preparation Example 3 albang dongsani Production Example 4 iron cage Preparation Example 5 Drungbang Dongsani Preparation Example 6 white room dongsani Preparation Example 7 rich man

Example 1. Confirmation of tyrosinase inhibitory activity of plant extracts of the genus Cyperus

To measure Tyrosinase inhibitory activity, mushroom tyrosinase, which is widely used in general, was used. A solution of 1 mM L-tyrosine and 0.5 M phosphate buffer (pH 6.5) is prepared in a 96-well microplate (SPL Life Sciences Co., Ltd., Pocheon, Korea). Extract samples to be measured were reacted with 170 μl of the prepared solution and 20 μl of tyrosinase (300 U) at 25° C. for 30 minutes. After the reaction, it was measured at 450 nm using a microplate reader (Berthold, Bad Wildbad, Germany). 10 μl of DMSO was added and used as a control. Blank was measured by adding 20 μl of 0.1 M phosphate buffer (pH 6.5) instead of tyrosinase.

The enzyme activity inhibition rate (%) was calculated by the following formula.

Tyrosinase activity inhibition rate (%) = {(A-B)/A} × 100

A: O.D at 450 nm without sample,

B: O.D at 450 nm with sample

division Absorbance relative to control
(Relative tyrosinase activity, Abs) Control (DMSO) 1.00 Preparation Example 1 0.314±0.05 Preparation Example 2 0.349±0.05 Preparation Example 3 0.428±0.05 Production Example 4 0.335±0.20 Preparation Example 5 0.337±0.2 Preparation Example 6 1.08±0.08 Preparation Example 7 0.97±0.12

As a result of the confirmation, as shown in Table 2, it can be seen that the inhibitory effect of tyrosinase is excellent in the groups treated with the extracts of five species of plants belonging to the genus Siperus of blue-bellied, gold-breasted, egg-breasted, brown-breasted, and drung-breasted. However, the efficacy was hardly shown in the extracts of the same genus, C. chinensis and Hyangbuja.

<Example 2. Confirmation of cytotoxicity of plant extract itself of the genus Cyperus>

HaCaT (Human ketatinocyte) was used to confirm the cytotoxicity of extracts of blue-bellied, gold-breasted, egg-breasted, brown-breasted-breasted, dreung-breasted-breasted, white-breasted-breasted-breasted-breasted-breasted-breasted-breasted-breasted. To this end, keratinocytes were cultured in a 96-well plate using DMEM (Dulbeco's Modified Eagle's Media) containing 10% (v/v) fetal bovine serum (FBS) for 18 hours, and keratinocytes without FBS were then cultured. It was replaced with sterile liquid medium 154CF for culture. Next, each well was treated with the extracts of Preparation Examples 1 to 7 at 100 μg/ml and cultured for 24 hours. Thereafter, 10 μl of CCK-8 reagent (Dojindo) was added per well, followed by incubation for 3 hours, and absorbance was measured at 450 nm. It was confirmed as shown in FIG. 1 at μg/ml, and it was confirmed that there was no cytotoxicity. However, among the extracts used as comparative conditions, the cytotoxicity of the cyanobacteria extract itself was confirmed.

Example 3. NO scavenging activity measurement test of plant extracts of the genus Cyperus

The NO scavenging ability of the extracts of Preparation Examples 1 to 7 was measured through the following method. Raw264.7 cells, mouse macrophages distributed from the Korea Cell Line Bank, were cultured in DMEM supplemented with 10% FBS and 1% penicillin-streptomycin at 37°C, 95% air, and 5% CO ₂ . It was seeded in a 96 well plate at 5×10 ⁴ cells/well. After each sample and positive control (quercetin 50 μM) were treated, they were cultured in an incubator for 2 hours, and each well was treated with LPS at a concentration of 1 μg/ml, and then cultured in an incubator for 24 hours. 50 μl of the supernatant was taken from the plate treated with the sample and LPS, put into a new plate, and the concentration of nitrite (NO ₂ ^- ) was measured and calculated for the production of NO. Nitrite concentration was measured by colorimetric assay using Griess reagent (1% sulfanilamide 0.1% napthylethylenediamine dihydrochloride / 2.5% H ₃ PO ₄ ). 50 μl of the culture solution was put in a 96 well plate, reacted with 50 μl of Griess reagent at room temperature in a light-shielded state for 10 minutes, and then the absorbance at 520 nm was measured using a microplate reader (Molecular Device, USA). The standard curve used the known concentration of sodium nitrite. The well containing only the culture medium was used as a control. Each experimental value was repeated at least three times and averaged.

Referring to Figure 2, it was confirmed that the extracts of Preparation Examples 1 to 5 significantly lowered the NO concentration in a concentration-dependent manner. ※ It seems that the white swallowtail extract of Preparation Example 6 also suppresses the NO concentration, but it is predicted that this is due to cell death, so the experiment is performed on the white swallowtail extract, but the effect is not explained.

Example 4. IL-6 confirmation experiment

Raw264.7 cells, mouse macrophages distributed from the Korea Cell Line Bank, were cultured in DMEM supplemented with 10% FBS and 1% penicillin-streptomycin at 37°C, 95% air, and 5% CO ₂ . After each sample was treated with 100 μg/ml and 200 μg/ml, each sample was incubated in an incubator for 2 hours, and each well was treated with LPS at a concentration of 1 μg/ml and cultured in an incubator for 24 hours. Take 50 μl of supernatant from the plate treated with sample and LPS, put it on a new plate, and measure the amount of IL-6 cytokine produced by a microplate reader (Beckman Coulter, CA, USA) using the IL-6 ELISA kit (Thermo Scientific, IL, USA). USA) was measured at a wavelength of 450 nm.

The IL-6 concentration is shown in FIG. 3, and the results also show that the extracts of Preparation Examples 1 to 5 have excellent efficacy in inhibiting IL-6 production.

Example 4. TNF-α confirmation experiment

Raw264.7 cells, mouse macrophages distributed from the Korea Cell Line Bank, were cultured in DMEM supplemented with 10% FBS and 1% penicillin-streptomycin at 37°C, 95% air, and 5% CO ₂ . After each sample was treated with 100 μg/ml and 200 μg/ml, each sample was incubated in an incubator for 2 hours, and each well was treated with LPS at a concentration of 1 μg/ml and cultured in an incubator for 24 hours. Take 50 μl of the supernatant from the plate treated with sample and LPS, put it on a new plate, and use the TNF-α ELISA kit (Cat # BMS223HS) to measure the amount of TNF-α produced using a microplate reader (Beckman Coulter, CA, USA) at 450 μl. It was measured at a wavelength of nm.

The TNF-α concentration is shown in Figure 4, and the results also show that the extracts of Preparation Examples 1 to 5 have excellent efficacy in inhibiting TNF-α production.

<Formulation Example 1. Cosmetics Manufacturing>

Formulation Example 1-1. manufacture of skins

5.0% by weight of the blue cockroach extract of Preparation Example 1 of the present invention, 5.2% by weight of propylene glycol, 1.5% by weight of oleyl alcohol, 3.2% by weight of ethanol, 3.2% by weight of polysorbate 20, 2.0% by weight of benzophenone-9, carboxyl A skin was prepared using a conventional method with the contents of 1.0% by weight of vinyl polymer, 3.5% by weight of glycerin, a trace amount of fragrance, a trace amount of preservative, and the remaining amount of purified water.

Formulation Example 1-2. manufacture of lotion

5.0% by weight of the goldenrod extract of Preparation Example 2 of the present invention, 1.0% by weight of cetostearyl alcohol, 0.8% by weight of glyceryl monostearate, 0.3% by weight of sorbitan monostearate, 1.0% by weight of polysorbate 60, mineral oil 5.0% by weight, 3.0% by weight of cyclomethicone, 0.5% by weight of dimethicone, 0.1% by weight of allantoin, 5.0% by weight of glycerin, 2% by weight of alcohol, 3.0% by weight of propylene glycol, a small amount of flavor, a small amount of preservative, and the remaining amount of purified water. The lotion was prepared using the phosphorus method.

Formulation Example 1-3. Preparation of Essence

5.0% by weight of eggplant extract of Preparation Example 3 of the present invention, 4.0% by weight of propylene glycol, 3.0% by weight of glycerin, 0.5% by weight of allantoin, 0.01% by weight of EDTA-2Na, 5.0% by weight of ethanol, 1.5% by weight of triethanolamine, 2.0% by weight of squalane %, 2.5% by weight of beeswax, 60% by weight of polysorbate, 1.0% by weight of carboxylvinyl polymer, 2.5% by weight of sorbitan sesquioleate, a small amount of flavor, a small amount of preservative, and the remaining amount of purified water. manufactured.

Formulation Example 1-4. preparation of cream

10.0% of the oxtail extract of Preparation Example 4 of the present invention, polyoxyethylene sorbitan monostearate 0.7%, sorbitan sesquioleate 0.5%, cetyl alcohol 0.6%, stearic acid 0.75%, glyceryl monostearate 0.6%, fluid A cream was prepared using a conventional method with the contents of 15.0% paraffin, 10.0% carboxyvinyl polymer, 0.2% triethanolamine, trace amounts of flavor, trace amounts of preservatives, and the remaining amount of purified water.

<Formulation Example 2. Food Manufacturing>

Formulation Example 2-1. Preparation of seasoning for cooking

A health-promoting cooking seasoning was prepared by adding 1% by weight of Dreongbangsani extract of Preparation Example 5 of the present invention to the cooking seasoning.

Formulation example 2-2. Manufacture of Flour Food

0.1% by weight of the blue cockerel extract of Preparation Example 1 of the present invention was added to wheat flour, and bread, cakes, cookies, crackers, and noodles were prepared using the mixture to prepare health-enhancing foods.

Formulation Example 2-3. Preparation of soups and gravies

Health-promoting soup and broth were prepared by adding 0.1% by weight of the goldenrod extract of Preparation Example 2 of the present invention to soup and broth.

Formulation Example 2-4. Manufacture of dairy products

The eggplant extract of Production Example 3 of the present invention was added to milk in an amount of 0.1% by weight, and various dairy products such as butter and ice cream were prepared using the milk.

Formulation Example 2-5. Preparation of vegetable juice

Vegetable juice for health promotion was prepared by adding 0.5 g of the extract of sardines in Preparation Example 4 of the present invention to 1,000 ml of tomato juice or carrot juice.

Formulation Example 2-6. manufacture of fruit juice

Fruit juice for health promotion was prepared by adding 0.1 g of the extract of Dreongbangsani of Preparation Example 5 of the present invention to 1,000 ml of apple juice or grape juice.

Claims (8)

A cosmetic composition for anti-inflammatory or skin whitening , characterized in that it contains an extract. delete A health functional food for anti-inflammatory or skin whitening, characterized in that it contains an extract of golden dongsani ( Cyperus microiria ). An anti-inflammatory composition comprising an extract of Cyperus microiria . A pharmaceutical composition for the prevention or treatment of inflammatory diseases , characterized in that it contains an extract. According to claim 5,
The inflammatory disease is selected from the group consisting of inflammatory bowel disease, inflammatory collagen vascular disease, glomerulonephritis, sarcoidosis, retinitis, gastritis, hepatitis, enteritis, arthritis, tonsillitis, sore throat, bronchitis, pneumonia, pancreatitis, cystitis, neuritis, sepsis and nephritis. A pharmaceutical composition for preventing or treating inflammatory diseases, characterized in that being. A health functional food for the prevention or improvement of inflammatory diseases, characterized in that it contains an extract of golden dongsani ( Cyperus microiria ). According to claim 7,
The inflammatory disease is selected from the group consisting of inflammatory bowel disease, inflammatory collagen vascular disease, glomerulonephritis, sarcoidosis, retinitis, gastritis, hepatitis, enteritis, arthritis, tonsillitis, sore throat, bronchitis, pneumonia, pancreatitis, cystitis, neuritis, sepsis and nephritis. Health functional food for preventing or improving inflammatory diseases, characterized in that being.

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