KR20230050744A - Composition for improving skin cooling sensation - Google Patents

Abstract

The present disclosure relates to a composition for enhancing skin cooling sensation comprising a Cnidium monnieri extract. The composition according to the present disclosure can enhance the skin cooling sensation by activating TRPM8, a cold receptor, without skin irritation.

Description

Composition for enhancing skin cooling sensation {COMPOSITION FOR IMPROVING SKIN COOLING SENSATION}

The present specification relates to a composition for enhancing skin cooling sensation.

A transient receptor potential (TRP) channel is a receptor that activates a cell by causing an influx of cations (eg, calcium ions) into the cell. TRPM8 (transient receptor potential melastatin 8), one of the transient receptor potential channels, is one of the temperature sensors present in the skin. TRPM8 is activated by cold stimulation at 20 to 26°C, which is slightly lower than body temperature, or by an agonist such as menthol to feel a cooling sensation.

In general, products that can feel a cooling sensation impart a cooling sensation by including ingredients such as menthol and mint. These methods are effective in imparting a cooling sensation, but menthol, mint, and the like are reverse stimulants and may cause burns, irritation, and pain, so their use in cosmetic compositions is limited. Therefore, a new composition capable of imparting a cooling sensation while minimizing skin irritation is required.

Menthol and related cooling compounds. Eccles R. J Pharm Pharmacol. 1994;46(8):618-30.

In one aspect, the present invention is to provide a composition that enhances the cooling sensation of the skin.

In one aspect, the present invention provides a composition for enhancing skin cooling sensation comprising a bee sandbox extract.

In an exemplary embodiment, the bee killer extract may be an extract of water, an organic solvent, or a mixed solvent thereof.

In an exemplary embodiment, the bee casualty extract may be an extract of 60 to 99.9% (w/w) ethanol aqueous solution.

In one exemplary embodiment, the bee casualty may be any one or more selected from the group consisting of leaves, stems and fruits of the bee casualty.

In one exemplary embodiment, the bee casualty extract may be an extract of the fruit of bee casualty.

In an exemplary embodiment, the bee casualty extract may be administered at a dosage of 600 to 6500 mg/kg/day.

In an exemplary embodiment, the composition may activate TRPM8 (transient receptor potential melastatin 8).

In an exemplary embodiment, the composition may be an external composition for skin.

In an exemplary embodiment, the composition may be a cosmetic composition.

In one aspect, the composition according to the present invention can improve skin cooling sensation without skin irritation.

In one aspect, the composition according to the present invention can activate TRPM8, a cold receptor.

1 is a graph showing the cytotoxicity of the bee casualty extract according to an embodiment of the present invention.
Figure 2 is a graph showing the TRPM8 activity of the bee casualty extract according to an embodiment of the present invention.
Figure 3 is a graph showing the mast cell degranulation inhibitory effect of the bee casualty extract according to one embodiment of the present invention.

Hereinafter, the present invention will be described in detail.

In one aspect, the present invention provides a composition for enhancing skin cooling sensation comprising an extract of Cnidium monnieri .

In the present specification, "bee casualty" refers to a biennial plant of the umbel family Miraniaceae, and the bee casualty has a straight stem, no hairs throughout, and leaves staggered and split into feathers three times. These bee casualties are distributed throughout the country as a native plant of Korea. The bee casualty may include any one or more selected from the group consisting of leaves, stems and fruits of the bee casualty.

In the present specification, the term "bee casualty extract" includes an extract of the bee killer, regardless of the extraction method, extraction solvent, extracted component or extract form, and includes a fraction obtained by fractionating the extracted extract with a specific solvent. The form of the extract may preferably be in the form of a powder. The extraction or fractionation may be performed using water, an organic solvent, or a mixed solvent thereof. Alcohol, isopropanol, acetone, hexane, ethyl acetate, carbon dioxide, or a mixed solvent of two or more of these may be used as the organic solvent, but is not limited thereto. can do. The alcohol may be a C ₁ ~ C ₅ lower alcohol. The number or method of extraction or fractionation is not particularly limited, and for example, methods such as cold extraction, ultrasonic extraction, reflux cooling extraction, and hot water extraction may be used. Preferably, the active ingredient is extracted or fractionated by cooling or heating After filtering, the filtrate is concentrated under reduced pressure to obtain the bee stinger extract of the present invention.

In one embodiment, the bee stippling extract may be an extract of water, an organic solvent, or a mixed solvent thereof. The water may include distilled water or purified water, and the organic solvent may be alcohol, isopropanol, acetone hexane, ethyl acetate, carbon dioxide, or a mixed solvent of two or more of these, but is not limited thereto. The alcohol may be a C ₁ -C ₅ lower alcohol.

In one embodiment, the bee casualty extract may be an extract of a mixed solvent of water and ethanol or an extract of ethanol. For example, the mixed solvent of water and ethanol is 60 to 99.9% (w / w) ethanol aqueous solution, 65 to 99.9% (w / w) ethanol aqueous solution, 65 to 95% (w / w) ethanol aqueous solution, 70 to 99.9% (w / w) ethanol aqueous solution 95% (w/w) ethanol aqueous solution, 70 to 90% (w/w) ethanol aqueous solution, 75 to 90% (w/w) ethanol aqueous solution, 75 to 85% (w/w) ethanol aqueous solution, 80 to 85% (w/w) ethanol aqueous solution or 80% (w/w) ethanol aqueous solution.

In one embodiment, the bee casualty extract by administration of the composition may be administered at a dosage of 600 to 6500 mg/kg/day. If the dose of the bee casualty extract is less than 600 mg/kg/day, the effect of enhancing skin cooling sensation is reduced, and if the dose exceeds 6500 mg/kg/day, irritation may occur. Specifically, the dosage of the bee casualty extract by administration of the composition is 600 mg / kg / day or more, 620 mg / kg / day or more, 640 mg / kg / day or more, 660 mg / kg / day or more, 680 mg /kg/day or more, 700 mg/kg/day or more, 720 mg/kg/day or more, 740 mg/kg/day or more, 760 mg/kg/day or more, 780 mg/kg/day or more, 800 mg/kg /day or more, 820 mg/kg/day or more, 840 mg/kg/day or more, 860 mg/kg/day or more, 880 mg/kg/day or more, 900 mg/kg/day or more, 950 mg/kg/day 1000 mg/kg/day or more, 1100 mg/kg/day or more, 1200 mg/kg/day or more, 1300 mg/kg/day or more, 1400 mg/kg/day or more, 1500 mg/kg/day or more; 1600 mg/kg/day or more, 1700 mg/kg/day or more, 1800 mg/kg/day or more, 1900 mg/kg/day or more, 2000 mg/kg/day or more, but less than or equal to 6500 mg/kg/day, 6480 Not more than mg/kg/day, not more than 6460 mg/kg/day, not more than 6440 mg/kg/day, not more than 6420 mg/kg/day, not more than 6400 mg/kg/day, not more than 6380 mg/kg/day, not more than 6360 mg/day kg/day or less, 6340 mg/kg/day or less, 6320 mg/kg/day or less, 6300 mg/kg/day or less, 6250 mg/kg/day or less, 6200 mg/kg/day or less, 6150 mg/kg/day or less day or less, 6100 mg/kg/day or less, 6000 mg/kg/day or less, 5900 mg/kg/day or less, 5800 mg/kg/day or less, 5700 mg/kg/day or less, 5600 mg/kg/day or less , 5500 mg/kg/day or less, 5400 mg/kg/day or less, 5300 mg/kg/day or less, 5200 mg/kg/day or less, 5100 mg/kg/day or less, 5000 mg/kg/day or less, 4900 or less than mg/kg/day, less than 4800 mg/kg/day, less than 4700 mg/kg/day, less than 4600 mg/kg/day or less than 4500 mg/kg/day.

In one embodiment, the composition may activate TRPM8 (transient receptor potential melastatin 8).

In one embodiment, the route of administration of the composition is not limited, but may be preferably applied to the skin.

In one embodiment, the composition may be a cosmetic composition. The cosmetic composition, for example, softening lotion, astringent lotion, nutrient lotion, nutrient cream, massage cream, eye cream, eye essence, essence, cleansing cream, cleansing lotion, cleansing foam, cleansing water, pack, powder, body lotion , Body cream, body essence, body cleanser, hair dye, shampoo, rinse, hair styling agent, hair growth agent, ointment, gel, cream, patch, spray, powder, and skin adhesive type, but may have formulations such as, but are not limited thereto.

In addition, other ingredients in addition to the essential ingredients described above in each formulation can be appropriately selected and blended without difficulty by those skilled in the art according to the type or purpose of use of other external preparations.

The cosmetic composition may be provided in any formulation suitable for topical application. For example, it may be provided in the form of a solution, an emulsion obtained by dispersing an oil phase in an aqueous phase, an emulsion obtained by dispersing an aqueous phase in an oil phase, a suspension, a solid, a gel, a powder, a paste, a microneedle, a foam, or an aerosol composition. there is. Compositions of such formulations may be prepared according to conventional methods in the art.

The cosmetic composition according to the present specification may further include functional additives and components included in general cosmetic compositions in addition to the compounds of the present specification. The functional additive may include a component selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high-molecular peptides, high-molecular polysaccharides, sphingolipids, and seaweed extracts. The cosmetic composition according to the present specification may include other components capable of giving a synergistic effect to the main effect within a range that does not impair the main effect. In addition, the cosmetic composition according to the present specification may further include a moisturizer, an emollient agent, a surfactant, a UV absorber, a preservative, a bactericide, an antioxidant, a pH adjuster, an organic and inorganic pigment, a fragrance, a cooling agent or an antiperspirant. The blending amount of the components can be easily selected by those skilled in the art within a range that does not impair the purpose and effect of the present specification, and the blending amount may be 0.001 to 10% by weight, specifically 0.01 to 3% by weight based on the total weight of the composition. there is.

Hereinafter, the present invention will be described in more detail through examples and the like. These examples are only for illustrating the present invention, and it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as being limited by these examples.

[Example 1] Preparation of bee casualty extract

Bee casualty fruit (Seoul, Korea) was dried and pulverized to obtain bee casualty powder. The powder was extracted with 80% (w/w) ethanol at 80° C. for 3 hours. The residue was extracted twice in the same manner, filtered, and then concentrated by vacuum evaporation to prepare a bee sage extract.

[Experimental Example 1] Measurement of cytotoxicity

Human epidermal keratinocytes (Human Epidermal Keratinocytes, LONZA) were added to KGM-Gold ^TM Keratinocyte Growth Medium (LONZA) and cultured under 37°C and 5% CO ₂ conditions. The cultured keratinocytes were inoculated into a 96 well plate at 20,000 cells/well, and then cultured for 24 hours at 37°C and 5% CO ₂ conditions. Here, the bee casualty extract of Example 1 was treated by concentration (10, 20, 50, 100 ppm) and cultured at 5% CO ₂ and 37 ° C for 24 hours. Then, after incubation with CCK-8 (DOJINDO Co.) for 2 hours, absorbance was measured at 450 nm using a microplate reader. In the measured results, relative cell viability was evaluated by setting the value of the control group to 100 and shown in FIG. 1 .

[Experimental Example 2] Evaluation of cooling sensation

TRPM8 (transient receptor potential melastatin 8) activation of bee casualty extract was evaluated using Fluo-4NW calcium assay. Fluo-4 AM is a fluorescent marker for measuring intracellular Ca ²⁺ concentration, and is converted into fluorescent Fluo-4 by esterification by esterase present in cells after permeating into cells. At this time, when Ca ²⁺ introduced into the cell through TRPM8 binds to Fluo-4, it emits fluorescence of a specific wavelength (Excitation/Emission: 494/516 nm). By measuring the fluorescence at each reaction time, the fluorescence increase value due to Ca ²⁺ influx is obtained, and through this, the relative increase in TRPM8 activity can be quantified.

CHO-K1 cells (Rugen SCI) overexpressing TRPM8 were cultured in RPMI-1640 (w/20mM HEPES, 10% FBS, 1% antibiotics) medium at 37°C and 5% CO ₂ . The cultured CHO-K1 cells were seeded on a 96 well black plate w/clear bottom at about 5x10 ⁴ /well, and then cultured in a cell culture medium for 24 hours. Using the Fluo-4NW calcium assay kit (F36206, Molecular probes), 100 ul of the dye solution was dispensed into the wells seeded with the CHO-K1 cells, and the bee killer extract of Example 1 was treated by concentration. Then, after blocking the light with foil and reacting for 30 minutes in a 37 ℃ incubator, it was taken out and reacted for another 30 minutes at room temperature. After the reaction, fluorescence was measured using a FlexStation device. The measured value was converted into (F-F0)/F0 and expressed as a bar graph, which is shown in FIG. 2. As a positive control group, Ionomycin was used to confirm whether the Fluo-4NW calcium assay reacted properly.

As shown in Figure 2, it was confirmed that the bee casualty extract enhances the cooling sensation by increasing the activity of TRPM8 in a concentration-dependent manner.

[Experimental Example 3] Mast cell degranulation inhibition evaluation

The mast cell degranulation inhibitory effect was confirmed by β-hexosaminidase assay. RBL-2H3 cells (22256, Korea Cell Line Bank) were cultured in DMEM medium (w/ 10% FBS, 1% antibiotics) at 37°C and 5% CO ₂ conditions. After seeding the cultured RBL-2H3 cells in a 96 well plate at 4x10 ⁴ /well, DNP-IgE (200ng/ml), fine dust (PM 2.5, 100ppm), and bee killer extract of Example 1 were treated by concentration. and cultured for 24 hours in a cell incubator. Fine dust was collected at a flow rate of 1.13 m ³ /min using a PTFE Teflon filter (Zefluor Co.) by installing a high volume sampler on the roof of Mizium, Amorepacific R&D Center. Then, the cells were washed twice with DPBS, treated with phenol red free DMEM (w/ 1% antibiotics) medium in which DNP-BSA was diluted to a concentration of 1 ug/ml, and cultured in an incubator for 1 hour. After the reaction, 50 ul of the supernatant from each well was transferred to a new 96 well plate, and 50 ul of substrate buffer (4-p-Nitrophenyl-N-acetyl-bD-glucosaminide 2 mM, sodium citrate 0.05 M, pH4.5) was added. Incubated for 1 hour in an incubator. After confirming that the color changed to yellow by adding 100 ul of stop solution per well, the absorbance was measured at 405 nm, and the control group (DNP-IgE + DNP-BAS) was converted to 100% in FIG. 3 and shown as a graph. Ketotifen was used as a positive control group.

As shown in Figure 3, it was confirmed that the bee casualty extract significantly inhibited mast cell degranulation.

[Formulation Example 1] Soap

Soap was prepared by a conventional method according to the composition shown in Table 1 below.

ingredient Content (% by weight) Bee casualty extract of Example 1 5.00 titanium dioxide 0.20 polyethylene glycol 0.80 glycerin 0.50 Ethylenediaminetetraacetic acid 0.05 salt 1.00 pigment 0.20 soap scent 0.20 soap base balance Sum 100.00

[Formulation Example 2] Lotion

A lotion was prepared in a conventional manner according to the composition shown in Table 2 below.

ingredient Content (% by weight) Bee casualty extract of Example 1 3.00 L-ascorbic acid-2-phosphate magnesium salt 1.00 Collagen 1.00 sodium citrate 0.10 citric acid 0.05 licorice extract 0.20 1,3-butylene glycol 3.00 Purified water balance Sum 100.00

[Formulation Example 3] Cream

Cream was prepared in a conventional manner according to the composition shown in Table 3 below.

ingredient Content (% by weight) Bee casualty extract of Example 1 5.00 Polyethylene glycol monostearate 2.00 Self-emulsifying monostearate glycerin 5.00 cetyl alcohol 4.00 squalene 6.00 Glyceryl tri-2-ethylhenate 6.00 sphingoglycolipids 1.00 1,3-butylene glycol 7.00 Purified water balance Sum 100.00

[Formulation Example 4] Pack

A pack was prepared in a conventional manner according to the composition shown in Table 4 below.

ingredient Content (% by weight) Bee casualty extract of Example 1 3.00 polyvinyl alcohol 13.00 L-ascorbic acid-2-phosphate magnesium salt 1.00 lauroylhydroxyproline 1.00 Collagen 2.00 1,3-butylene glycol 3.00 ethanol 5.00 Purified water balance Sum 100.00

[Formulation Example 5] Beauty liquid formulation

According to the composition shown in Table 5 below, a cosmetic liquid formulation was prepared by a conventional method.

ingredient Content (% by weight) Bee casualty extract of Example 1 3.00 Hydroxyethylene Cellulose 12.00 xanthan gum 2.00 1,3-butylene glycol 6.00 glycerin 4.00 sodium hyaluronate 5.00 Purified water balance Sum 100.00

Claims (9)

A composition for enhancing skin cooling sensation, comprising a bee burial extract. According to claim 1,
The bee casualty extract is an extract of water, an organic solvent, or a mixed solvent thereof, the composition. According to claim 1,
The bee casualty extract is an extract of 60 to 99.9% (w / w) ethanol aqueous solution. According to claim 1,
The bee casualty is any one or more selected from the group consisting of leaves, stems and fruits of the bee casualty, the composition. According to claim 1,
The bee casualty extract is an extract of the fruit of the bee casualty, composition. According to claim 1,
The bee casualty extract is administered at a dosage of 600 to 6500 mg / kg / day, composition. According to claim 1,
Wherein the composition activates TRPM8 (transient receptor potential melastatin 8). According to any one of claims 1 to 7,
The composition is a composition for external application to the skin. According to any one of claims 1 to 7,
The composition is a cosmetic composition, the composition.

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