Anti-inflammatory, skin-soothing, irritation-relieving, and skin barrier-improving cosmetic composition containing cyperus rotundus extract

Abstract

The present invention relates to a cosmetic composition containing a Cyperus rotundus extract extracted by heating microbubbles. The extract can inhibit the generation of NO (Nitric Oxide), can inhibit the expression of inflammatory expression factors, i.e., interleukin-6 (IL-6), cyclooxygenase-2 (COX-2), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α), thereby exhibiting excellent anti-inflammatory effects, can strengthen the expression of skin barrier and moisturizing genes, i.e., Aquaporin-3 (AQP-3), Hyaluronan Synthase 2 (HAS-2), claudin-1 (CLDN-1), Cadherin 1 (CDH1), Filaggrin (FLG), and involucrin (IVL), thereby exhibiting excellent effects in skin soothing, moisturizing, skin barrier improvement, and the relief of irritation such as skin redness, and the relief of itching, etc. when used as a cosmetic, and thus can be effectively used as a multi-care product for improving the skin condition.

Description

Technical Field

[0001] This invention relates to a cosmetic composition containing Cyperus rotundus extract manufactured by a microbubble extraction process, which has anti-inflammatory, skin-soothing, irritation-relieving, and skin barrier-improving effects. Background Technology

[0002] Cyperus rotundus is a perennial herb belonging to the Cyperaceae family, growing in southern Korea and Jeju Island. It contains essential oil components such as pinene, campene, 1,8-cineol, and limonene, and possesses analgesic and antibacterial properties. However, using existing solvents for extraction via static extraction may limit the application of the effective components in Cyperus rotundus.

[0003] Microbubbles can achieve functions such as sterilization and washing through their physicochemical properties such as gas dissolution, self-pressurization, and electrification, as well as the redox effect of ultrafine large bubbles and the large amount of energy generated when the bubbles burst. Therefore, they are widely used in various fields.

[0004] Recently, we have been actively developing and applying products and services with unique microbubbles, thereby gradually expanding their application areas. They are widely used in all industries such as food, medical, agriculture, aquaculture and dyeing. Moreover, as described in Korean Patent No. 10-2222661, microbubbles can also be used for the extraction of natural raw materials.

[0005] Therefore, in the process of studying the various physiological activities of Cyperus rotundus, the inventors confirmed that the Cyperus rotundus extract extracted by microbubbles has better anti-inflammatory, skin-soothing, irritation-relieving and skin barrier-improving effects than the existing static extracts, and can be used as a new cosmetic, thus completing the present invention.

[0006] Prior technology documents

[0007] Patent documents

[0008] (Patent Document 1) Republic of Korea Patent Publication No. 2013-0062112 (Title of Invention: Topical Skin Composition for Prevention and Treatment of Inflammatory Diseases Containing Cyperus rotundus Extract as an Active Ingredient; Applicant: Korea Pharmaceutical Industries Promotion Agency; Publication Date: June 12, 2013.)

[0009] (Patent Document 2) Republic of Korea Patent Registration No. 10-2222661 (Title of Invention: Method for Manufacturing Extract of Natural Cosmetic Raw Material Using Heated Microbubbles and Cosmetic Composition Containing the Extract; Applicant: J2KBIO Co., Ltd.; Registration Date: August 24, 2020.)

[0010] (Patent Document 3) U.S. Patent No. 11110117 (Title of Invention: Skin preparation composition for external use containing complex hyaluronic acid, Applicant: J2KB IO Co., Ltd., Registration Date: 2021.09.07) Summary of the Invention

[0011] The purpose of this invention is to provide a cosmetic composition containing an extract of Cyperus rotundus produced by a microbubble extraction process, which has anti-inflammatory, skin-soothing, irritation-relieving, and skin barrier-improving effects.

[0012] This invention relates to a cosmetic composition for anti-inflammatory and skin-soothing purposes, containing an extract of Cyperus rotundus extracted by heating microbubbles.

[0013] The extract may contain 50,000 to 60,000 ppm of α-cyperone.

[0014] Preferably, the extract is characterized by inhibiting the production of NO (nitric oxide). Furthermore, it also has the effect of inhibiting the expression of genes selected from the group consisting of inflammatory expression factors namely interleukin-6 (IL-6), cyclooxygenase-2 (COX-2), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α).

[0015] The extract may have skin barrier improving effects. Furthermore, the extract may have moisturizing effects by inhibiting transepidermal water loss and increasing skin moisture content. Preferably, the extract is characterized by enhancing the expression of genes selected from the group consisting of aquaporin-3 (AQP-3), hyaluronic acid synthase-2 (HAS-2), human claudin-1 (CLDN-1), cadherin 1 (CDH1), filaggrin (FLG), and involucrin (IVL).

[0016] Furthermore, the extract can have skin irritation-relieving effects. Preferably, the extract can relieve skin irritation and soothe the skin by relieving skin erythema or improving itching symptoms.

[0017] The heated microbubble extraction can be performed using a microbubble generator with a pore size of 5–50 μm and an output rate (or liquid flow rate) of 1.0–1.2 Ton / hr, and extraction with an organic solvent at 40–60 °C for 3–5 hours. At this time, the microbubble generation conditions are a pressure of 3–4 bar and… Its extraction efficiency is optimal under the specified gas flow rate conditions.

[0018] The organic solvent may be a C1-C4 alcohol or an aqueous solution thereof. The C1-C4 alcohol may be methanol, ethanol, propanol, isopropanol, butanol, and isobutanol, and the aqueous solution of the C1-C4 alcohol may be a 40-60% (v / v) aqueous solution of the C1-C4 alcohol.

[0019] The organic solvent can be used at 1 to 40 times its volume based on the weight of Cyperus rotundus used (based on a 1 kg volume). ).

[0020] The extracts of the present invention may include fractions thereof. As used herein, the term "fraction" refers to a product obtained by fractionation performed to separate a specific component or group of components from a mixture comprising multiple constituent parts.

[0021] The fractionation method for obtaining the fractionated product in this invention is not particularly limited and can be performed according to methods commonly used in the relevant art. Examples include solvent fractionation, which involves processing with multiple solvents; ultrafiltration fractionation, which involves passing the product through an ultrafiltration membrane with a specific molecular weight cutoff; chromatographic fractionation, which involves performing various chromatography methods (for separation based on size, charge, hydrophobicity, or affinity); and combinations thereof. The type of fractionation solvent used to obtain the fractionated product of this application is not particularly limited, and any solvent known in the relevant art can be used. Non-limiting examples of the fractionation solvent in this application include polar solvents such as water and alcohols with 1 to 4 carbon atoms; and non-polar solvents such as hexane, ethyl acetate, chloroform, and dichloromethane. One or more solvents can be used alone or in combination.

[0022] The chromatography method can be selected from silica gel column chromatography, LH-20 column chromatography, ion exchange resin chromatography, medium pressure liquid chromatography, thin layer chromatography (TLC), silica gel vacuum liquid chromatography, and high performance liquid chromatography.

[0023] Furthermore, this invention provides a pharmaceutical composition containing a Cyperus rotundus extract extracted by heated microbubbles, which has anti-inflammatory, skin-soothing, irritation-relieving, and skin barrier-improving effects. The Cyperus rotundus extract may be added to the pharmaceutical composition of this invention at a concentration of 0.001–100% by weight.

[0024] The pharmaceutical composition can be formulated into oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, or into external forms such as suppositories and sterile injectable solutions, using conventional methods. Carriers, excipients, and diluents that can be included in the pharmaceutical composition can include, for example, lactose, glucose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylparaben, propylparaben, talc, magnesium stearate, and mineral oil. During formulation, commonly used diluents or excipients such as fillers, expanders, humectants, disintegrants, and surfactants can be used. Solid dosage forms for oral administration include tablets, alkyl groups, powders, granules, and capsules. As described above, these solid dosage forms can be prepared by adding at least one excipient, such as starch, calcium carbonate, sucrose, lactose, and gelatin, to the Cyperus rotundus extract of this invention. Furthermore, in addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. Liquid dosage forms for oral administration include suspensions, oral liquids, emulsions, and syrups. Besides commonly used simple diluents such as water and liquid paraffin, they can also contain various excipients such as wetting agents, sweeteners, flavoring agents, and preservatives. Dosage forms for non-oral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions can be propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Bases for suppositories can include Wipedol, Macrogol, Tween 61, cocoa butter, glyceryl laurate, and glycerin gelatin.

[0025] The dosage of the pharmaceutical composition of the present invention can vary depending on the age, sex, and weight of the recipient, the specific disease or pathological condition to be treated, the severity of the disease or pathological condition, the route of administration, and the prescribing physician's judgment. Those skilled in the art can determine the dosage based on the factors described above, according to their own expertise; typical dosages range from 0.01 mg / kg / day to approximately 2000 mg / kg / day. More advanced dosages range from 1 mg / kg / day to 500 mg / kg / day. Administration can be once daily or divided into multiple administrations. The dosages described are not intended to limit the scope of the invention in any way.

[0026] The pharmaceutical compositions of the present invention can be administered to mammals such as mice, livestock, and humans via various routes. Administration can be performed by any means, such as oral, rectal or intravenous, intramuscular, subcutaneous, intradural, or intracerebral injection. Because the Cyperus rotundus extract of the present invention has virtually no toxicity or side effects, it is a safe medicine even for long-term use for preventative purposes.

[0027] Furthermore, this invention provides a health functional food containing a Cyperus rotundus extract extracted by heated microbubbles and food additives that are food-grade and have anti-inflammatory, skin-soothing, irritation-relieving, and skin barrier-improving effects. The Cyperus rotundus extract can be added to the health functional food of this invention at a rate of 0.001–100% by weight. The health functional food of this invention includes forms such as tablets, capsules, alkyl forms, or liquids. Foods to which the extract of this invention can be added include various beverages, meats, sausages, bread, candies, snacks, noodles, ice cream, dairy products, soups, ionic beverages, drinking water, alcoholic beverages, chewing gum, tea, and vitamin complexes.

[0028] As a dosage form of the cosmetic composition of the present invention containing Cyperus rotundus extract extracted by heating microbubbles, it can be manufactured in any dosage form commonly manufactured in the industry, and may include, for example, serums, lotions, emulsions, face masks, hand creams, foot creams, lip balms, lipsticks, eyeshadows, eyeliners, eyebrow pencils, blushes, highlighters, general toners, skin lotions, face creams, serums, beauty soaps, softening toners, medicated toners, body washes, facial foams, facial lotions, gels, facial oils, facial creams, shampoos, hair dyes, hair conditioners, hair conditioners, facial cleansing sheets, and facial water.

[0029] Specifically, when the dosage form of the cosmetic composition according to the present invention is a paste, cream, or gel, the carrier component may include animal fibers, plant fibers, waxes, paraffin wax, starch, astragalus gum, cellulose derivatives, polyethylene glycol, silicon, bentonite, silica, talc, or zinc oxide. When the dosage form of the cosmetic composition according to the present invention is a powder or spray, the carrier component may include lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder. Particularly in the case of a spray, a propellant such as chlorofluorocarbons, propane, or dimethyl ether may be added. When the dosage form of the cosmetic composition according to the present invention is a solution or emulsion, the carrier component may include solvents, solvators, or emulsifiers, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylethylene glycol oil, glycerol aliphatic esters, polyethylene glycol, or fatty acid esters of sorbitan anhydride. When the dosage form of the cosmetic composition according to the present invention is a suspension, liquid diluents such as water and ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol and polyoxyethylene sorbitol ester, microcrystalline cellulose, aluminum hydroxide, bentonite, agar, or astragalus gum can be used as carrier components. When the dosage form of the cosmetic composition according to the present invention is a surfactant-containing cleanser, carrier components such as fatty alcohol sulfates, fatty alcohol ether sulfates, sulfosuccinate monoesters, acesulfonates, imidazoline derivatives, methyl taurate, sarcosinates, fatty acid amide ether sulfates, alkyl amide betaine, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolenic acid derivatives, or ethoxylated glycerol fatty acid esters can be added. The cosmetic composition according to the present invention may be supplemented with excipients including fluorescent substances, bactericides, water-soluble growth promoters, moisturizers, fragrances, fragrance carriers, proteins, solubilizers, sugar derivatives, sunscreens, vitamins, and plant extracts. The amount of the ingredient added can be selected according to the dosage form or intended use, within a range that does not impair the inherent effects of the cosmetic composition. For example, the amount of the ingredient added relative to the total weight of the composition can be 0.1 to 10% by weight, preferably 0.1 to 6% by weight, but is not limited thereto.

[0030] This invention relates to a cosmetic composition containing an extract of Cyperus rotundus extracted by heated microbubbles. The extract can inhibit the production of nitric oxide (NO) and suppress the expression of inflammatory factors, namely interleukin-6 (IL-6), cyclooxygenase-2 (COX-2), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α), thus exhibiting excellent anti-inflammatory effects. It can also inhibit the expression of skin barrier and moisturizing genes, namely aquaporin-3 (AQP-3), hyaluronic acid synthase-2 (HAS-2), human claudin-1 (CLDN-1), and cadherin-1 (CDH1). 1) Enhanced expression of filaggrin (FLG) and involucrin (IVL) results in remarkable effects when used in cosmetics, such as skin soothing, moisturizing, skin barrier improvement, relief of skin irritation such as erythema, and relief of itching. Therefore, it can be effectively used as a multi-care product to improve skin condition. Attached Figure Description

[0031] Figure 1 This is a chart showing the results of a face cream containing the Cyperus rotundus extract extracted by heating microbubbles according to the present invention, demonstrating its itch-relieving effect.

[0032] Figure 2 This is a chart showing the results of a face cream containing the Cyperus rotundus extract extracted by heated microbubbles according to the present invention, demonstrating its effect in inhibiting transdermal water loss.

[0033] Figure 3 This is a chart showing the results of a face cream containing the Cyperus rotundus extract extracted by heating microbubbles according to the present invention, which has the effect of increasing skin moisture content. Detailed Implementation

[0034] The preferred embodiments of the present invention will now be described in detail. However, the present invention is not limited to the embodiments described herein, but can be implemented in other forms. The embodiments described are merely for the purpose of more thoroughly and completely introducing the content of the present invention and fully conveying the ideas of the present invention to those skilled in the art.

[0035] <Examples 1 to 3: Preparation of Cyperus rotundus microbubble extract>

[0036] Cyperus rotundus root was purchased from Samhonggeonjae Pharmaceutical (Korea). After microbubble extraction, 100g of Cyperus rotundus root and 2000g of a 50% (v / v) ethanol aqueous solution were injected into a microbubble generator (manufacturer: AutoBubble) with pore sizes of 5–50 μm and an outflow rate (liquid flow rate) of 1.0–1.2 Ton / hr. The mixture was then filtered using a 5 μm pore size filter (No. 2, HYUNDAIMICRO), followed by concentration, drying, and vacuum drying, and used as a sample.

[0037] Furthermore, in the microbubble generator, the pressure at which bubbles are generated using air from the atmosphere is 3–4 bar, and the gas flow rate of the air is [missing information].

[0038] Table 1

[0039]

[0040] <Comparative Examples 1 to 13: Preparation of Comparative Extracts>

[0041] To manufacture Cyperus rotundus extract, wherein the extraction method is modified to the conditions described below.

[0042] Table 2

[0043]

[0044] <Experimental Example 1: Identification of Active Ingredients in Various Extracts>

[0045] A standard of α-cyperone, the indicator component of the extract, was purchased from Sigma-Aldrich. 100 mg of the standard was dissolved in… Dissolve 50 mg of each sample in 99% methanol. In 99% methanol.

[0046] The analytical system used was a Waters e2695 / 2998UVD, and the column used was a Capcellpak C18 UG120 (5 μm, 4.6 × 250 mm). For the elution gradient, solvent A was a 0.1% aqueous formic acid solution, and solvent B was acetonitrile (ACN). Elution was performed in 13 fractions over 80 minutes, from solvent A to solvent B, at a ratio of 100:0 (v:v) → 0:100 (v:v).

[0047] The content of the effective components in each sample is shown in Table 3 below.

[0048] Table 3

[0049] extract α-Cyperone (ppm) Example 1 53241.41 Example 2 51434.03 Example 3 50463.39 Comparative Example 1 40454.07 Comparative Example 2 42523.03 Comparative Example 3 34253.35 Comparative Example 4 33423.09 Comparative Example 5 34523.01 Comparative Example 6 42234.23 Comparative Example 7 40234.02 Comparative Example 8 35523.42 Comparative Example 9 35245.30 Comparative Example 10 39283.43 Comparative Example 11 34234.52 Comparative Example 12 34526.35 Comparative Example 13 35422.34

[0050] The results in Table 3 confirm that the extracts of Examples 1 to 3 of the present invention contain the highest content of the effective compound, namely α-cyperone.

[0051] <Experimental Example 2: Confirmation of Cytotoxicity of the Sample>

[0052] To confirm the cytotoxicity of the various extracts prepared in Examples 1 to 3 and Comparative Examples 1 to 13, a cell viability analysis (MTT assay) was performed. Cells were first analyzed in 96-well plates at 5 × 10⁻⁶ cells / well. 4 Mouse mononuclear macrophage leukemia cells (RAW 264.7) were inoculated at cell / well concentrations, followed by the addition of samples dissolved in dimethyl sulfoxide (DMSO) at various concentrations after 24 hours. After 24 hours, the culture medium was removed, and 40 μL of MTT reagent (in PBS 2.5 mg / mL) was added to each well, incubated for 4 hours, and then the supernatant was removed. The cells were then completely dissolved by adding 100 μL of DMSO. The absorbance at 540 nm was measured using a spectrophotometer.

[0053] Table 4

[0054]

[0055] The results are shown in Table 4. Assuming the control group value is 100%, the individual extracts up to... At the concentrations observed up to this point, cell viability of over 97.0% was observed, indicating no cytotoxicity; therefore, the next experiment was conducted... Implement at the following concentrations.

[0056] <Experimental Example 3: Determination of NO Generation>

[0057] To confirm the inhibitory activity of NO (nitric oxide) production in the extracts prepared in Examples 1 to 3 and Comparative Examples 1 to 13, experiments were conducted using mouse macrophage cells, specifically the mouse mononuclear macrophage leukemia cell line (RAW 264.7). The assay was first performed in 96-well plates at a concentration of 1.0 × 10⁻⁶. 4Mouse mononuclear macrophage leukemia cells (RAW264.7) were inoculated at a cell / well concentration and then cultured for 24 hours in a 37°C incubator supplied with 5% CO2.

[0058] Next, the 96-well plates were treated with staged dilutions of the sample and a stimulant, with the final concentration of lipopolysaccharide (LPS) at 1 μg / mL. 100 μL of the supernatant from cells cultured for 24 hours was transferred to a new 96-well plate. Griess A and Griess B solutions were mixed in a 1:1 ratio and 100 μL was added to each well. The reaction was carried out at room temperature for 10 minutes, and the absorbance at 540 nm was measured.

[0059] The nitric oxide production rate was calculated using the formula shown below.

[0060] NO generation rate (%) = (sample absorbance / reference absorbance) × 100

[0061] To visualize the results, the optical density (OD) values ​​of cells treated with 1 μg / mL lipopolysaccharide (LPS) were converted to 100%, and the results were recorded as a control group.

[0062] Table 5

[0063]

[0064] The results are shown in Table 5. It can be confirmed that the extracts of Examples 1 to 3 of the present invention exhibit a concentration-dependent NO generation inhibition effect. However, the extracts that did not meet the standard could not effectively inhibit NO generation, and therefore their anti-inflammatory efficacy was low.

[0065] <Experiment 4: Investigation of Anti-inflammatory Related Gene Expression Levels>

[0066] First, in a 96-well plate, at a depth of 4.0 × 10⁻⁶ mm. 5 Mouse mononuclear macrophage leukemia cells (RAW 264.7) were inoculated at cell / well concentrations and then cultured for 24 hours in a 37°C incubator supplied with 5% CO2. Individual extract samples were treated to a final concentration of 100 ppm and reacted with 1 μg / mL lipopolysaccharide (LPS) as a stimulant for 4 hours.

[0067] After removing the supernatant from the culture medium and washing with phosphate-buffered saline (PBS), the sample was dissolved in 500 μL of NucleoZOL (Macherey-Nagel, Germany). Then, 200 μL of sterile distilled water was added, and the sample was centrifuged for 15 minutes. The supernatant was then transferred to a new 500 μL tube. Next, 500 μL of isopropanol was added, and the sample was centrifuged for 10 minutes. After removing the supernatant, 75% (v / v) aqueous ethanol was added, and the sample was centrifuged again for 5 minutes. After removing the supernatant and drying at room temperature, the sample was dissolved in 50 μL of diethyl pyrocarbonate-distilled water (DEPC-DW), and the ribonucleic acid (RNA) was quantified using a spectrophotometer.

[0068] Next, 1 μg of ribonucleic acid (RNA) quantified using the HiSenScript™ RH(-)RT PreMix Kit and ethylene glycol monocarbonate-distilled water (DEPC-DW) were added to bring the total volume to 20 μL. Complementary deoxyribonucleic acid (cDNA) was synthesized at 42°C for 30 min, 85°C for 10 min, and 4°C at ∞. The synthesized cDNA was then mixed with 10 μL of 2x Real-Time PCR Master Mix (BioFACT), 1 μL of forward primer (10 pmol / μL), 1 μL of reverse primer (10 pmol / μL), and ethylene glycol monocarbonate-distilled water (DEPC-DW) to bring the total volume to 20 μL. The primers used to confirm the target gene were those disclosed in Table 6 below.

[0069] Table 6

[0070]

[0071] Next, polymerase chain reaction (PCR) was performed using a real-time polymerase chain reaction (RTPCR) device under the condition of 50 cycles at 95°C for 15 minutes, 95°C for 20 seconds, and 60°C for 40 seconds. The results were analyzed by relative quantitative analysis (ΔCt value) to determine the expression of messenger ribonucleic acid (mRNA) of each factor.

[0072] The expression levels of each gene are shown in Table 7 below.

[0073] Table 7

[0074]

[0075] The results are shown in Table 7. It can be found that the expression of each gene of inflammatory expression factors, namely interleukin-6 (IL-6), cyclooxygenase-2 (COX-2), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α), was significantly inhibited in the extract treatment groups of Examples 1 to 3.

[0076] <Experimental Example 5: Confirmation of the expression levels of skin barrier and moisturizing-related genes>

[0077] Next, to confirm the skin barrier strengthening effect, samples were taken from 6-well plates at a density of 2.0 × 10⁻⁶ mm. 5 Keratinocytes (HaCaT) were seeded at a cell / well concentration and cultured for 24 hours in a 37°C incubator supplied with 5% CO2. After washing with phosphate-buffered saline (PBS) and replacing with Duchenne Modified Igor Medium (DMEM) without fetal bovine serum (FBS), each extract sample was treated to a final concentration of 100 ppm. RNA was extracted and cDNA synthesized under the same assay conditions as for anti-inflammatory genes, and gene expression assays for aquaporin-3 (AQP-3), hyaluronic acid synthase-2 (HAS-2), human claudin-1 (CLDN-1), cadherin 1 (CDH1), filaggrin (FLG), and involucrin (IVL) were performed in the same manner. At this point, the primers used are shown in Table 8 below, and the gene expression results are output first in Table 9 with the comparison group (untreated group) as 100%.

[0078] Table 8

[0079]

[0080] Table 9

[0081]

[0082] As shown in Table 9, the analysis of gene expression patterns of aquaporin-3 (AQP-3), hyaluronic acid synthase-2 (HAS-2), human claudin-1 (CLDN-1), cadherin 1 (CDH1), filaggrin (FLG), and involucrin (IVL), which are related to skin barrier strengthening and moisture / moisturization, confirms that the extracts of Examples 1 to 3 of the present invention are most suitable for use as cosmetic compositions that strengthen the skin barrier by enhancing skin moisturizing function.

[0083] <Example 1 of cosmetic composition formulation: Manufacturing of skin lotion>

[0084] A skin lotion was prepared using the extract of Example 1 (5.0 wt%), propylene glycol (5.2 wt%), oleyl alcohol (1.5 wt%), ethanol (3.2 wt%), polysorbate 20 (3.2 wt%), benzophenone-9 (2.0 wt%), carboxyvinyl polymer (1.0 wt%), glycerin (3.5 wt%), trace amounts of fragrance, trace amounts of preservative, and the remainder of purified water, according to the usual method.

[0085] <Example 2 of cosmetic composition formulation: Manufacturing of moisturizing lotion>

[0086] A moisturizing lotion was prepared using the extract of Example 1 (5.0 wt%), cetearyl alcohol (1.0 wt%), glyceryl monostearate (0.8 wt%), sorbitan monostearate (0.3 wt%), polysorbate 60 (1.0 wt%), mineral oil (5.0 wt%), cyclomethicone (3.0 wt%), dimethicone (0.5 wt%), allantoin (0.1 wt%), glycerin (5.0 wt%), alcohol (2 wt%), propylene glycol (3.0 wt%), trace amounts of fragrance, trace amounts of preservatives, and the remainder of purified water, according to the usual method.

[0087] <Example 3 of cosmetic composition formulation: Manufacturing of toner>

[0088] A toner was prepared using the extract from Example 1 (5.0 wt%), propylene glycol (4.0 wt%), glycerin (3.0 wt%), allantoin (0.5 wt%), disodium EDTA-2Na (0.01 wt%), ethanol (5.0 wt%), triethanolamine (1.5 wt%), squalane (2.0 wt%), beeswax (2.5 wt%), polysorbate (60 wt%), carboxyvinyl polymer (1.0 wt%), sorbitol sesquioleate (2.5 wt%), trace amounts of fragrance, trace amounts of preservatives, and the remainder of purified water, according to the usual method.

[0089] <Example 4 of cosmetic composition formulation: Manufacturing of face cream>

[0090] The cream was prepared using the extract from Example 1 (5.0 wt%), polyoxyethylene sorbitan monostearate (0.7 wt%), sorbitan sesquioleate (0.5 wt%), cetyl alcohol (0.6 wt%), stearic acid (0.75 wt%), glyceryl monostearate (0.6 wt%), liquid paraffin (15.0 wt%), carboxyvinyl polymer (10.0 wt%), triethanolamine (0.2 wt%), trace amounts of fragrance, trace amounts of preservatives, and the remainder of purified water, according to the usual method.

[0091] <Experiment 6: Confirmation of Skin Irritation Relief Effect>

[0092] A human patch test was conducted using the toner manufactured in cosmetic formulation example 3 to evaluate its skin irritation relief effect.

[0093] Therefore, 20 adult women aged 20 to 55 were selected as subjects (Day -1, the start date of the experiment). After cleaning the test area, the skin was soothed for 30 minutes, and then an evaluation was carried out using a spectrophotmeter.

[0094] At this point, a chamber containing a 1.0% sodium lauryl sulfate (SLS) aqueous solution was applied to the test site for 24 hours to induce skin erythema. The condition of the test site was then assessed after removing the chamber (day 0 before product use).

[0095] The toner was applied to test subjects who were induced to develop erythema as described above and whose condition was measured. Safety and equipment evaluations were conducted on the test subjects 24, 48, and 72 hours after product use, thereby evaluating its efficacy.

[0096] The results of confirming the *a value using a colorimeter (Spectrophotometer) show that the *a value significantly decreased after 72 hours of product use.

[0097] Table 10

[0098]

[0099] As shown in Table 10, in the cases where sodium lauryl sulfate (SLS) patches were used alone, the redness and irritation of the skin reached their maximum after 24 hours. However, in the cases where product patches containing Cyperus rotundus extract obtained in Example 1 were used, the skin condition recovered rapidly. However, the toner without the extract still left red residue after 72 hours.

[0100] <Experimental Example 7: Expert Visual Assessment>

[0101] The face cream of Formulation Example 4 (containing 5% by weight of Cyperus rotundus extract from Example 1) was compared with other face creams containing the same ingredients but without the extract to test its clinical efficacy. In this study, the same subjects were observed until the user evaluation.

[0102] Therefore, after cleansing twice daily (morning and evening) for eight weeks, various face creams were applied to areas within 5cm below the knee creases of subjects who typically experienced itching. A total of 32 subjects participated in the trial.

[0103] First, experts (dermatologists / professional physicians) visually assess the degree of erythema, keratinization, induration, and fissuring based on the amount of face cream used, according to the ESIF scale (erythema, scaling, duration, fissuring) in Table 11 below.

[0104] Table 11

[0105]

[0106] Through expert evaluation, the four variables mentioned above can be added together, that is, the total score of each test subject can be 0 to 12 points. If the score exceeds 6 points, the subject can be determined to need dermatological treatment and excluded from the test.

[0107] The results confirmed that, among the 32 participants over a total of 8 weeks, none had a combined score exceeding 6 for the variables of erythema, keratosis, induration, and cracking. No individuals were observed to be unsuitable for the trial or experiencing side effects; therefore, the trial continued. These results were not presented separately.

[0108] <Trial Example 8: Confirmation of Improvement in Itching Symptoms>

[0109] Itching symptoms were assessed in subjects undergoing an 8-week trial using the Visual Analogue Scale (VAS). The length of each 10cm line segment marked directly by the subjects was measured. The rate of change in itching severity was calculated using the following formula. *Before product use: Week 0

[0110] Change rate in itching severity (%)

[0111] = {(Intensity of itching after product use - Intensity of itching before product use) / Intensity of itching before product use} × 100

[0112] The result is as follows Figure 1 As shown.

[0113] pass Figure 1 It can be confirmed that the face cream containing Cyperus rotundus extract (TEST) showed a significant relief of itching symptoms in weeks 4 and 8 compared to the face cream in the control group (CONTROL) without extract treatment.

[0114] <Experimental Example 9: Confirmation of Transdermal Water Loss and Skin Moisture Content>

[0115] Transepidermal water loss (TEWL), measured according to the level of application of face cream, is tested in accordance with the Human Suitability Testing Guidelines for Cosmetics that Help Improve Itching Symptoms by Restoring Skin Barrier Function, <Publicly Accepted Guidelines>. Transepidermal water loss is measured using a Vapometer (Delfin, Phinland). The Vapometer is equipped with a chamber that measures the relative humidity of the skin surface, thus measuring the amount of water lost from the skin surface. The measured value decreases as transepidermal water loss (TEWL) decreases, and the unit of measurement is g / m³. 2 / h, the result calculated using the following formula is as follows: Figure 2 As shown. *Before using the product: Week 0

[0116] Change rate of skin moisture content (%)

[0117] = {(Measurement value after product use - Measurement value before product use) / Measurement value before product use} × 100

[0118] pass Figure 2 It can be confirmed that the face cream containing Cyperus rotundus extract (TEST) significantly reduced transepidermal water loss in weeks 4 and 8 compared to the face cream in the control group (CONTROL) without extract treatment.

[0119] <Experimental Example 10: Confirmation of Skin Moisture Content>

[0120] The skin moisture content, based on the amount of face cream used, was measured using a CM825 skin moisture meter (Courage-Khazaka electronic GmbH, Germany). The skin moisture meter measures the capacitive capacitance of the current transmitted through a probe in contact with the skin. Because moisture content and capacitive capacitance are directly proportional, higher moisture levels result in higher measured values. The measurement coefficient is in arbitrary units (AU). The results are converted using the following formula: Figure 3 As shown. *Before using the product: Week 0

[0121] Change rate of skin moisture content (%)

[0122] = {(Measurement value after product use - Measurement value before product use) / Measurement value before product use} × 100

[0123] See Figure 3 In the group that applied a face cream containing the Cyperus rotundus extract of the present invention, the skin moisture content increased significantly.

[0124] <Experiment Example 11: Subject Satisfaction>

[0125] According to the criteria in Table 12 below, user satisfaction with the face cream containing the Cyperus rotundus extract of the present invention and the control group was measured after 4 weeks, and the results are shown in Table 13.

[0126] Table 12

[0127] Fraction Dosage form standard Standards for improvement of itching symptoms 1 Very bad The overall itching symptoms worsened significantly. 2 Difference The overall itching symptoms worsened 3 generally No difference compared to before application 4 excellent Overall itching symptoms improved 5 Excellent The overall itching symptoms were significantly improved.

[0128] Table 13

[0129]

[0130] The results in Table 13 confirm that the face cream containing the extract of the present invention not only improves itching symptoms, but also has excellent affinity for the cosmetic formulation itself.

[0131] The results confirm the effectiveness of the face cream containing the Cyperus rotundus extract of the present invention.

Claims

1. A cosmetic composition for anti-inflammatory and skin-soothing purposes, characterized in that: Contains Cyperus rotundus extract extracted using heated microbubbles. The heated microbubble extraction uses pores with a size of 5 to 50. A microbubble generator with a discharge rate of 1.0–1.2 Ton / hr is used, and extraction with an organic solvent is performed at a temperature of 40–60°C for 3–5 hours. The microbubble generator generates microbubbles at a pressure of 3–4 bar and a gas flow rate of 3–4 bar. / min, wherein the organic solvent is an aqueous solution of a C1-C4 alcohol at 40-60% v / v.

2. A cosmetic composition for anti-inflammatory and skin-soothing purposes, characterized in that: Contains Cyperus rotundus extract extracted using heated microbubbles. The heated microbubble extraction uses pores with a size of 5 to 50. A microbubble generator with a discharge rate of 1.0–1.2 Ton / hr was used, and extraction was performed at 70°C using an organic solvent for 4 hours. The microbubble generator produced microbubbles at a pressure of 3 bar and a gas flow rate of 4... / min, wherein the organic solvent is a 60% v / v aqueous solution of ethanol.

3. The anti-inflammatory and skin-soothing cosmetic composition according to claim 1 or 2, characterized in that: It contains α-cyperone.

4. The anti-inflammatory and skin-soothing cosmetic composition according to claim 1 or 2, characterized in that: The extract inhibits the formation of nitric oxide.

5. The anti-inflammatory and skin-soothing cosmetic composition according to claim 1 or 2, characterized in that: The extract inhibits the expression of genes selected from the group consisting of interleukin-6, cyclooxygenase-2, interleukin-1β, and tumor necrosis factor-α, which are inflammatory expression factors.

6. The anti-inflammatory and skin-soothing cosmetic composition according to claim 1 or 2, characterized in that: The extract has skin barrier improving effects.

7. The anti-inflammatory and skin-soothing cosmetic composition according to claim 1 or 2, characterized in that: The extract enhances the expression of genes selected from the group consisting of aquaporin-3, hyaluronic acid synthase-2, human blocking protein-1, cadherin 1, polyfilamentin, and inner lining protein.

8. The anti-inflammatory and skin-soothing cosmetic composition according to claim 1 or 2, characterized in that: The extract has the effect of relieving skin irritation.

9. The anti-inflammatory and skin-soothing cosmetic composition according to claim 8, characterized in that: The skin irritation is characterized by symptoms such as erythema or itching.

10. The anti-inflammatory and skin-soothing cosmetic composition according to claim 1, characterized in that: The temperature is 40°C, 50°C, or 60°C.

11. The anti-inflammatory and skin-soothing cosmetic composition according to claim 1, characterized in that: The time is 3 hours, 4 hours or 5 hours.

12. The anti-inflammatory and skin-soothing cosmetic composition according to claim 1, characterized in that: The pressure is 3 bar or 4 bar.

13. The anti-inflammatory and skin-soothing cosmetic composition according to claim 1, characterized in that: The gas flow rate is 3 / min or 4 / min.

14. The anti-inflammatory and skin-soothing cosmetic composition according to claim 1, characterized in that: The organic solvent is an aqueous solution of a C1-C4 alcohol at 40% v / v, 50% v / v, or 60% v / v.

15. The anti-inflammatory and skin-soothing cosmetic composition according to claim 1 or 14, characterized in that: The C1 to C4 alcohols are methanol, ethanol, propanol, isopropanol, butanol, or isobutanol.

16. The anti-inflammatory and skin-soothing cosmetic composition according to claim 15, characterized in that: The C1 to C4 alcohols mentioned are ethanol.

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