KR101477959B1 - Anti-irritating cosmetic compositions for skin comprising extracts of Aloe Aborescens - Google Patents

Abstract

The present invention relates to a skin irritation resistant cosmetic composition including an aloe extract, a Magnolia denudata Desrousseaux extract, and a peppermint extract as active ingredients. The skin irritation resistant cosmetic composition includes the aloe extract, Magnolia denudata Desrousseaux extract, and peppermint extract at a weight ratio of 1:1:1. The skin irritation resistant cosmetic composition decreases the generation of COX-2 and eotaxin-1, activates a cold sensor, and increases a moisturizing change rate of skin so that inflammatory skin diseases can be alleviated or treated. Moreover, the cosmetic composition blocks moisture loss of damaged skin or alleviates erythema and exhibits an effect in refreshing skin. Accordingly, if the cosmetic composition is used as an external application agent, skin damaged by external irritation can be soothed. In addition, the cosmetic composition does not have cell toxicity and skin side effects so as to be stably applied to a cosmetic composition.

Description

[0001] The present invention relates to a cosmetic composition for skin anti-irritation comprising an aloe extract as an active ingredient,

The present invention relates to a cosmetic composition for skin anti-irritation comprising an aloe extract, a Shin extract and a peppermint extract as an active ingredient. More particularly, the present invention relates to a cosmetic composition for skin anti-irritation, which comprises an anti-inflammatory, moisturizing, The present invention relates to a cosmetic composition for safe skin anti-irritation when applied to human body.

Skin is stimulated by retinol and lactic acid which are widely used as raw materials for cosmetics including various pollutants such as environmental pollution and automobile exhaust gas, and skin diseases such as aging, skin inflammation such as atopic or acne and wrinkles are increasing .

The major cause of skin diseases is known to be caused by inflammation in the body. Nitric oxide (NO), a kind of reactive radical species in the body inflammatory reaction, is produced by immune cells such as macrophages, It is known to play an important role in the pathological process. The nitrogen monoxide is generated through the activation of iNOS (inducible nitric oxide synthase), a nitric oxide (NO) generating enzyme by an inflammatory substance such as lipopolysaccaride (LPS) as a very unstable compound. COX-1 and COX-2 are known as COX (cyclooxygenase), which is also known as PGHS (prostaglandin H synthase), and is an important regulatory enzyme that acts to convert membrane phospholipids into prostalandins (thromboxanes) . COX-1 is basically present in tissues and constantly exhibits a certain action, whereas COX-2 is expressed in inflammatory reaction sites in animals or humans. It is known to be rapidly expressed in a short period of time by various stimuli. (COX-2) expression is also known to affect the production of nitrogen monoxide (NO) (Daewoo, Journal of the Korean Society of Cosmetic Scientists (2006) 58: 173-180). NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) activates an inflammatory gene expression mechanism to activate tumor necrosis factor (TNF), interleukin-1 interleukin-1, IL-1, interleukin-6, IL-6, interleukin-8 and IL-8, granulocyte macrophage-colony stimulating factor, GM-CSF), inducible nitric oxide synthase (iNOS), intercellular adhesion molecule 1 (ICAM-1), E-selectin, MHC class I and II molecules It is known as a transcription factor that increases expression. Thus, inhibitors of NF-κB may be effective in treating chronic and acute inflammatory diseases and atopic dermatitis.

It is also known that various types of cytokines or chemokines are involved in the process of infiltrating inflammatory cells into the lesion, and related receptors are expressed in various cells such as inflammatory cells and keratinocytes. Chemokine is a family of cytokines, now known as more than 50 species. They play a role in attracting appropriate immune cells to the site while the tissue is causing an inflammatory reaction. In particular, atopic dermatitis plays an important role in the infiltration of inflammatory cells into the skin. RANTES (regulated after activation, normal T expressed and secreted), monocyte chemotactic protein 4 (MCP4) Thaxin-1 is frequently elevated in atopic dermatitis lesions, which are closely related to the chemotaxis of eosinophils with CCR3 receptors and Th2 cells. Eosinophils secrete highly toxic protein and free radicals that can kill microbes and parasites, but on the other hand they can be very tightly regulated as they can cause serious tissue damage to the host if they are inadequately activated Loses. These eosinophils are Eotaxin-1, which mediates the transfer of circulating blood to tissues, and Eotaxin-1 modulation is a new target for atopic treatment.

On the other hand, in general, when the low-temperature environment is formed in the skin, the threshold value of inflammation-induced pain is lowered. It can be applied to reduce the inflammation and the pain caused by applying ice padding to skin inflammation sites or similar low temperature environment. TRPM-8 (transient receptor potential cation channel subfamily M member 8, cold and menthol-sensitive receptor 1: CMR1) is a type of transient receptor potential (TRP) ion channel that responds to cold stimulation. It is activated by the temperature of 25 ℃ or less due to the characteristic of TRPM8 ion channel. It is also activated by chemicals such as menthol, menthone, eucalyptol, spearmint, icilin (Reid G, Ion channels activated by cold and menthol in cultured rat dorsal root ganglion neurons. Neurosci Lett 2002; 324: 164-8). Recent studies have shown that TRPM8 is a new target for anti-inflammation, which can alleviate inflammation and reduce the feeling of lethargy, as well as coolness effects (Ramachandran R. TRPM8 activation attenuates inflammatory responses in mouse models of colitis, Proc Natl Acad Sci U.S.A. 2013 Apr 30; 110 (18): 7476-81).

Currently, steroid preparations, topical immunosuppressants, topical antibiotics and antihistamines are used as medicines for treating inflammatory skin diseases. However, when the medicinal preparations are used for a long period of time, skin thinning, discoloration of skin color, osteoporosis, arteriosclerosis, And the like. In addition, in the case of skin inflammation, various internal inflammatory signals are involved, and skin barrier weakness exacerbates such inflammation.

For this reason, a multi-functional substance capable of simultaneously exhibiting a anti-inflammatory effect for relieving inflammatory skin disease, a cold effect for reducing inflammation and pain, and a moisturizing effect for improving skin barrier, Is being emphasized.

Under these circumstances, the inventors of the present invention have made efforts to develop a natural-derived compound as a composite material having anti-inflammatory, cold and moisturizing effects, and high stability and safety, .

One object of the present invention is to provide a cosmetic composition for skin anti-irritation comprising an aloe extract, a Shin extract and a peppermint extract as an active ingredient.

The present invention relates to a cosmetic composition for skin anti-irritation comprising an aloe extract, a Shin extract and a peppermint extract as an active ingredient.

In the present invention, the cosmetic composition for skin anti-irritation refers to a cosmetic composition containing the anti-cancer effect, the cold feeling effect and the moisturizing effect at a weight ratio higher than that of aloe extract, Shin extract or peppermint extract, respectively. Specifically, the cosmetic composition for skin anti-irritation according to the present invention contains aloe extract, Shin extract and peppermint extract in a weight ratio of 0.9-1.1: 0.9-1.1: 0.9-1.1, preferably 1: 1: 1 Is included.

In one specific embodiment, an aloe extract, a god extract mixing peppermint extract in various weight ratios by preparing a complex compound, and the intracellular calcium by those of NF-κB luciferase activity, the TRFM8 activity (Ca ^{2 +)} influx and When used as an external preparation for skin, the moisture resistance was measured. As a result, the complex compound prepared by mixing aloe extract, Shin extract and peppermint extract at various weight ratios showed that the activity of NF-κB luciferase was at least 30% or more, preferably 40 , More preferably at least 50%, even more preferably at least 60%, still more preferably at least 70%, and the content of calcium (Ca ²⁺ ) introduced during TRPM8 activation is determined by the amount of TRPM8 not activated More preferably at least 100% by weight, more preferably at least 150% by weight, based on the weight of calcium introduced into the group, and when the composition is used as an external preparation for skin, By at least 5%, preferably at least 10%, more preferably at least 15%, even more preferably at least 25%.

In addition, the compound (# 1) prepared by mixing Aloe extract, Shin extract and Mint extract in a weight ratio of 1: 1: 1 among the complex compounds prepared according to the weight ratio, It was confirmed that NF-κB luciferase activity was remarkably inhibited compared to other complex compounds (# 4, # 6 and # 9) prepared by mixing, and aloe extract having the best moisturizing effect was mixed at a weight ratio of 1.5 or more It was confirmed that the amount of moisturizing change was remarkably smaller than that of the complex compound (# 2, # 3 and # 8). The complex compound (# 1) prepared by mixing Aloe extract, Shin extract and Mint extract at a weight ratio of 1: 1: 1 was mixed with 2 parts by weight of the mint extract having the best cold effect, (# 5 and # 7) prepared by mixing the peppermint extract at a weight ratio of 1.5 or more, the amount of calcium introduced into the cells was significantly increased compared to that of the compound (# 5 and # 7) (See FIGS. 1, 2 and 3).

Thus, the cosmetic composition in which the aloe extract, Shin extract and peppermint extract of the present invention are contained at a specific weight ratio, that is, a weight ratio of 0.9-1.1: 0.9-1.1: 0.9-1.1, preferably 1: 1: 1, Inhibits related immune related reactions, activates cold sensation sensors, and increases the amount of moisturizing change of the skin. Therefore, it can improve or treat inflammatory skin diseases, and when used as a external preparation, it gives a feeling of cooling sensation to damaged skin, It has the purpose of blocking loss or improving erythema.

The inflammatory skin disease may be, but is not limited to, at least one selected from the group consisting of atopic dermatitis, seborrheic dermatitis, contact dermatitis, irritable lupus, urticaria, acne and psoriasis.

Hereinafter, aloe extract, Shin extract and peppermint extract, which are effective components of the composition of the present invention, are referred to as a complex compound (ASAP-calm).

In one specific embodiment, the complex compound (ASAP-calm) has a content of COX-2 (cyclooxygenase-2) produced by LPS stimulation of at least 5%, preferably at least 10%, more preferably at least 20% , More preferably at least 30%, even more preferably at least 50%, even more preferably at least 60%, and the content of eotaxin-1 produced by interleukin stimulation is at least 15% or more, preferably 50% or more, more preferably 80% or more, still more preferably 90% or more. In addition, when the external preparation for skin including the complex compound is applied to the skin damaged by external stimuli, the TEWL is cut by 20 to 35% as compared with a case where the external preparation containing no compound is applied, 15 to 40%, and a cooling sensation is felt (see Figs. 4, 5, 6 and 10).

As used herein, the term " anti-irritation "is meant to include skin soothing as the condition of the skin damaged or improved by external stimuli is improved or improved. Preferably, the inflammation-related skin inflammation-related immune reaction is inhibited, thereby improving or treating the inflammatory skin disease, cooling effect that can reduce inflammation and pain, and moisturizing effect to improve skin barrier, Improvement or treatment.

As used herein, the term " improvement "or" treatment "means (a) inhibition of the development of a disease or disease in an individual, (b) relief of the disease or disorder, and (c) As used herein, the term "individual" means a mammal, including a human, having a disease whose symptoms may be ameliorated by administration of the composition of the present invention, such as a monkey, cow, horse, pig, sheep, dog, cat, rat, Means a mammal.

In the present invention, the aloe extract, Shiney extract and peppermint extract can be obtained by using an extraction method and an extraction solvent known in the art. The extraction solvent may be selected from the group consisting of water, methanol, ethanol, isopropanol, butanol, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, dichloromethane, N, N-dimethylformamide, dimethylsulfoxide (DMSO) , 1,3-butylene glycol, propylene glycol, or a mixed solvent thereof. The extract can be extracted at room temperature or with heating under conditions where the active ingredient of the extract is not destroyed or minimized. Depending on the extraction solvent to be extracted, the degree of extraction and the degree of loss of the active ingredient of the extract may differ. Therefore, an appropriate extraction solvent should be selected and used. The extraction method is not particularly limited, and examples thereof include cold extraction, ultrasonic extraction, and reflux cooling extraction.

In addition, the aloe extract, Shin-Ii extract and peppermint extract may be obtained through a conventional purification process in addition to the extraction method using the above-mentioned extraction solvent. For example, various purification processes have been carried out, including ultrafiltration with a constant molecular weight cut-off value, freezing filtration, centrifugation, various chromatography (size, charge, hydrophobicity or affinity chromatographic separation) But the present invention is not limited thereto.

In the present invention, the aloe extract, Shiney extract and peppermint extract all include all extracts, fractions and tablets obtained in each step of extraction, fractionation or purification, their diluted solutions, concentrates or dried products.

In the present invention, the cosmetic composition may contain components commonly used in cosmetic compositions in addition to the complex compound. For example, conventional auxiliaries acceptable as cosmetics such as antioxidants, stabilizers, solubilizers, thickeners, vitamins, pigments and flavors, and carriers.

The cosmetic composition may be prepared in any formulations conventionally produced in the art. For example, they may be formulated as solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansers, oils, powder foundations, emulsion foundations, wax foundations and sprays, But is not limited thereto. More specifically, it can be prepared as a flexible lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.

When the formulation of the cosmetic composition is a paste, cream or gel, an animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, Can be used.

When the cosmetic formulation is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, chlorofluorohydrocarbons, propane / Propane, such as butane or dimethyl ether.

When the cosmetic composition is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid esters of sorbitan, and the like.

When the formulation of the cosmetic composition is in the form of a suspension, the carrier component may be a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tragacanth.

When the formulation of the cosmetic composition is an interfacial active agent-containing cleansing, it is preferable to use, as carrier components, aliphatic alcohol sulfates, aliphatic alcohol ether sulfates, sulfosuccinic acid monoesters, isethionates, imidazolinium derivatives, methyltaurates, sarcosinates, fatty acids Amide ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester.

The active ingredient of the complex compound prepared by mixing the aloe extract, Shin extract and peppermint extract of the present invention is 0.0001 to 20% by weight, preferably 0.001 to 10% by weight, more preferably 0.01 to 10% by weight, 3% by weight. When the weight of the complex compound is less than 0.0001% by weight, the skin soothing effect is weak. When the weight of the compound is more than 20% by weight, the effect of increasing the content is very weak.

Meanwhile, as a result of the cytotoxicity and skin irritation test of the complex compound in the concrete examples of the present invention, it has been found that the complex compound is a cytotoxic substance to keratinocytes as a natural substance and a harmless substance to human body. Accordingly, the complex compound prepared by mixing the aloe extract, Shin extract and the peppermint extract of the present invention has little toxicity and side effects, so that it can be safely used even for long-term use. In particular, as described above, the cosmetic composition for skin anti- Can be applied.

The cosmetic composition for skin anti-irritation according to the present invention contains aloe extract, Shin extract and peppermint extract at a weight ratio of 1: 1: 1, which reduces COX-2 production and eotaxin-1 production, , It is possible to improve or treat inflammatory skin diseases by increasing the amount of moisturizing change of the skin. In addition, the cosmetic composition has the effect of blocking water loss of damaged skin, improving erythema and refreshing feeling on the skin. Therefore, when used as an external preparation, the skin damaged by external stimuli can be calm. In addition, since it is free from cytotoxicity and skin side effects, it can be safely applied to a cosmetic composition.

FIG. 1 is a graph showing the effect of inhibiting NF-.kappa.B luciferase activity of complex compounds prepared by mixing Aloe extract, Shin extract and Mint extract according to the composition ratio.
FIG. 2 is a graph showing Ca ²⁺ influx into cells of complex compounds prepared by mixing Aloe extract, Shin extract and Mint extract according to the composition ratio.
FIG. 3 is a graph showing a change in moisture change of composite compounds prepared by mixing Aloe extract, Shin extract and Mint extract according to a composition ratio.
FIG. 4 shows the inhibitory effect of COX-2 on the expression of COX-2 in astaxanthin cells according to the concentration of the complex compound in which aloe extract, Shin extract and peppermint extract were mixed at a weight ratio of 1: 1: 1 Graph.
FIG. 5 shows the effect of reducing the secretion of eotaxin-1 according to the concentration of a complex compound in which aloe, extract, and peppermint extract were mixed at a weight ratio of 1: 1: 1 when eotaxin-1 was induced in human normal fibroblasts Fig.
FIG. 6 is a graph showing the results of water loss (TEWL) and erythema (redness) in the case of applying a composition prepared by concentration of a complex compound in which aloe extract, Shin extract and peppermint extract were mixed at a weight ratio of 1: 1: erythema).

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention, and it is to be understood by those skilled in the art that the present invention is not limited thereto It will be obvious.

Example 1: Selection of natural products having anti-inflammation, cold feeling and moisturizing effect

Based on the results of previous studies of the present inventors, 20 kinds of extracts having anti-inflammatory activity were selected, and the efficacy of these extracts was compared with anti-inflammatory, cold sensation and moisturizing parts.

1-1. Manufacture of natural extract

The plant parts shown in the following Table 1 were collected and washed thoroughly so as to completely remove impurities and impurities, shaken at 20 to 35 ° C, and pulverized to a particle size of 1 mm or less. Then, 100 g of the pulverized extract powder was immersed in 10 L of 70% ethanol and subjected to ultrasonic extraction for 24 hours. The obtained extract was then filtered through a filter paper (Advantes, No. 2) and concentrated under reduced pressure to prepare each extract.

1-2. Anti-inflammatory effect test

Each of the plant extracts obtained in Example 1-1 was tested for anti-inflammatory activity by inhibiting NF-κB luciferase activity. Specifically, NF-κB reporter DNA was inserted into human fibroblasts and NIH / 3T3 cells, a mouse fibroblast cell line, using Superfect, and the transformation was induced. Twenty-four hours after transformation, the samples were treated with TNF-alpha (10 ng / ml). After lapse of 16 hours, the cells were collected and luminescence was measured at 450 nm using a luminometer (Berthold, Germany) (see: Planta medica 2005; 71: 338-343). The results are shown in Table 1.

1-3. Cold feeling efficacy test

In order to examine the activity of TRPM8 (transient receptor potential cation channel subfamily M member 8) in each of the plant extracts obtained in Example 1-1, Ca ²⁺ influx was measured to be Ca ²⁺ 2 / AM (Molecular probe, F36205), which specifically binds and reacts. Specifically, HEK cells in which hTRPM8 was cloned (SB-HEK-hTRPM8, sb ion channel) were cultured at 2 × 10 ⁴ cells in 96 wells for fluorescence microscopy. The next day, 5 μM of Fura-2 / AM prepared in HBS balanced salt solution was added and incubated at room temperature for 45 minutes. Cells that were then loaded with Fura-2 / AM were transferred to a measuring device with a confocal microscope (IN CELL ANALYZER 1000, GE Healthcare, USA). Cells were excited at 360 and 380 nm (F340, F380) and fluorescence was measured at 510 nm. Images were stored every 4 seconds and the amount of calcium in the cells was expressed as the relative ratio of F340 and F380. Cooling efficacy was judged based on Ca ²⁺ influx and persistence. The results are shown in Table 1.

1-4. Moisturizing effect black

Moisture level of plant extracts was measured using MPA9 (corneometer, CK electronic, Germany) for the measurement of transepidermal water loss (TEWL) of each plant extract extracted in Example 1-1. The plant extracts were applied to the abdomen upper part of the experimenter after applying the same amount of patches (adhesive plaster, Ripafet Pharma, Japan) and waited for 30 minutes. Then, after the patch was removed, the degree of moisturization at each application site was measured at regular time intervals, and the amount of moisturizing change was calculated using the following equation (1). All measurements were carried out in a constant temperature and humidity room without air movement and without direct sunlight at room temperature of 22 ° C and relative humidity of 40%. The results are shown in Table 1.

[Equation 1]

Moisture change (%) = {(measured value after patch removal - measured value after applying plant extract) / measured value after patch removal} × 100

Extraction name (used site) Anti-inflammatory effect
(NF-kB) Moisturizing effect Cold feeling effect Selection Scarlet fever (fruit) ++ + - Chrysanthemum (chrysanthemum) (flower) +++ + - Forsythia ++ - - King Cherry tree (branches) +++ - - Crowd (outpost) ++ - - Mint (Outpost) +++ - +++ O Shin (Magnolia flower) ++++ ++ - O Mulberry (fruit) ++ + - Strawberry (Outpost) +++ - + Aloe leaves +++ ++++ + O Camphor tree +++ - ++ Eucalyptus (leaf) + + ++ Peppermint (Outpost) ++ - ++ Mountain Mint (Outpost) ++ - ++ Oatmeal (fruit) + +++ - Basil seeds (fruit) + +++ - Pepper (fruit) + - ++ Chestnut trees (branches and leaves) +++ + + Iris (outpost) ++ + - Mustard (Outpost) ++ - ++ -: no effect, +: weak effect, ++: moderate effect, +++: strong effect ++++: very strong effect

As a result of the test, 20 kinds of plant extracts showed the highest efficacy among anti-inflammatory, moisturizing, and cold sensation effects. Among the extracts that showed the highest efficacy among the other evaluation items, aloe, cinnamon, Respectively. These compounds were tested for their ability to exert their combined action on sedation and synergistic effects.

Example 2: Establishment of composition ratio of a compound showing anti-inflammatory, moisturizing, and cold effect

2-1. Composite compound manufacturing

The aloe extract, Shin extract and Mint extract obtained in Example 1 were mixed at various ratios to prepare a complex compound. The composition ratio of the composite compound was prepared in ten composition ratios as shown in Table 2 below.

Composite number Aloe extract Shin extract Peppermint extract One One One One 2 1.5 One 0.5 3 1.5 0.5 One 4 One 1.5 0.5 5 One 0.5 1.5 6 0.5 1.5 One 7 0.5 One 1.5 8 2 0.5 0.5 9 0.5 2 0.5 10 0.5 0.5 2

2-2. Measurement of anti-inflammatory effect of compound

The anti-inflammatory effect of each compound prepared in Example 2-1 was measured in the same manner as in Example 1-2. As a control, NF-κB reporter DNA-transfected fibroblasts were treated with TNF-α and fibroblasts not treated with complex compound, and positive control cells were transformed with NF-κB reporter DNA Fibroblasts treated with TNF-α alone were used for the fibroblasts. The results are shown in FIG. 1 and Table 3.

Complex compound number NF-κB activity Standard Deviation NF-κB (% of cont) Standard deviation (% of cont) Control group 67 4 25% One% Positive control group
(TNF? (100 ng / ml)) 273 8 100% 3% # 1 (100 ppm) 75 5 27% 2% # 2 (100 ppm) 147 3 54% One% # 3 (100 ppm) 169 7 62% 3% # 4 (100 ppm) 83 12 30% 4% # 5 (100 ppm) 139 10 51% 4% # 6 (100 ppm) 110 11 40% 4% # 7 (100 ppm) 105 9 38% 3% # 8 (100 ppm) 186 12 68% 4% # 9 (100 ppm) 111 4 41% One% # 10 (100 ppm) 162 6 59% 2%

As a result, the cells treated with the compound (# 1) prepared by mixing Aloe extract, Shin extract and Mint extract at a weight ratio of 1: 1: 1 showed NF-κB luciferase activity as a 73% inhibition (# 4), which was mixed at a weight ratio of 1: 1.5: 0.5 with the compound (# 4) prepared at a weight ratio of 0.5: 1: 1.5, Inhibition of NF-κB luciferase activity by 70% and 62%, respectively.

2-3. Measurement of cold effect of composite compound

The cooling effect of each compound prepared in Example 2-1 was measured in the same manner as in Example 1-3. As a control, HEK cells cloned with hTRPM8 without complex compound treatment were used. Cooling efficacy was judged based on Ca ²⁺ influx and persistence. The results are shown in FIG. 2 and Table 4.

Complex compound number 10 seconds 30 seconds 60 seconds 90 seconds 120 seconds Control group 12 13 9 10 11 # 1 (100 ppm) 25 150 178 180 177 # 2 (100 ppm) 20 78 83 85 72 # 3 (100 ppm) 26 102 121 125 111 # 4 (100 ppm) 23 71 79 73 64 # 5 (100 ppm) 24 112 125 130 128 # 6 (100 ppm) 22 83 80 81 77 # 7 (100 ppm) 25 125 145 155 150 # 8 (100 ppm) 20 78 83 85 72 # 9 (100 ppm) 21 78 83 85 72 # 10 (100 ppm) 32 152 193 194 189

As a result, Ca ^{2 +} influx was highest at 189 RFU in cells treated with compound (# 10) prepared by mixing Aloe extract, Shin extract and Mint extract at a weight ratio of 0.5: 0.5: 2, (177 RFU) treated with a complex compound (# 1) prepared by mixing at a weight ratio of 2: 1: 1, a cell treated with a complex compound (# 7) mixed at a weight ratio of 0.5: RFU), 1: 0.5: 1.5 are mixed in a weight ratio of the process the complex compound produced (# 5) cells (128 RFU) were confirmed to be highly pure influx of Ca ^{+ 2.}

2-3. Measurement of moisturizing effect of compound

The moisturizing effect of each compound prepared in Example 2-1 was measured in the same manner as in Example 1-4. As a control group, patches without complex compound treatment were used. After removing the patch, the degree of moisturization at each application site was measured at regular intervals of time, and the amount of moisturizing change was calculated using Equation 1. The results are shown in FIG. 3 and Table 5.

Complex compound number 0 minutes 30 minutes Moisture change (%) Standard Deviation Control group 82 24 71% 5% # 1 (0.1%) 81 44 46% 4% # 2 (0.1%) 83 43 48% 3% # 3 (0.1%) 82 38 54% 2% # 4 (0.1%) 80 31 61% 4% # 5 (0.1%) 81 29 64% 3% # 6 (0.1%) 79 27 66% 3% # 7 (0.1%) 78 30 62% One% # 8 (0.1%) 82 41 50% 4% # 9 (0.1%) 80 30 63% 2% # 10 (0.1%) 81 29 64% 4%

As a result of the experiment, when the patch containing the complex compound prepared according to the composition ratio was applied, the amount of moisturizing change was included in the composite compound (# 1) prepared by mixing Aloe extract, Shin extract and Mint extract at a weight ratio of 1: 1: 1 (# 2), which was prepared by mixing at a weight ratio of 1.5: 0.5: 1, was 48%, the moisture change was 48% : 0.5: 0.5 weight ratio, the amount of water change was 50% when a patch containing the composite compound (# 8) prepared as described above was applied.

In general, the combination of aloe extract, Shin extract and peppermint extract showed higher efficacy than single extract alone, and there was a significant correlation between composition ratio and efficacy , But it was confirmed that the composition ratio showing the ideal synergistic effect in the anti-inflammatory effect, moisturizing effect, and cold feeling effect was mixed with the composition ratio of aloe extract, Shin extract and peppermint extract of 1: 1: 1 (Aloe: Shini: peppermint).

Hereinafter, the complex compound is referred to as ASAP-calm.

Example  3: Cytotoxicity test of compound

The stability of the most effective compound (ASAP-calm) revealed in Example 2 was examined. Specifically, cytotoxicity test (MTT) was performed using HaCaT cells, which are human keratinocyte cell lines. HaCaT cells were treated with the complex compound (ASAP-calm) of Example 2 at the concentrations of 0, 10, 25, 50, 100, 250, 500, 1,000 and 2,000 占 퐂 / The cytotoxic effect was measured using MTT (3- (4,5-dimetylthiazol-2-yl) 2,5-diphenyltetrazolium bromide) assay. The results are shown in Table 6.

The concentration (쨉 g / ml) of the complex compound (ASAP-calm) 0 10 25 50 100 250 500 1,000 2,000 Cell survival rate 100 101 98.7 102 99.3 97.4 95.8 87.9 79.8

As a result, the complex compound (ASAP-calm) did not show cytotoxicity at a concentration of 500 μg / ml or less, but it was confirmed that some cells were killed at a concentration of 1,000 μg / ml or more.

Example  4: COX -2 generation inhibitory effect measurement

Immunized macrophages (RAW 264.7) were cultured in 24-well plates for 24 hours, transfected with COX-2 luciferase vector, and cultured for 24 hours. The cells were treated with LPS (Lipopolysaccharide) at a concentration of 50 ppm, 100 ppm and 300 ppm, and the cells were treated with 200 ng / ml of LPS to induce COX-2 production. The degree of COX-2 production was then measured using the degree of luciferase fluorescence expression. The results are shown in Fig.

As a result, the production of COX-2 was induced when LPS was administered to immune macrophages and the production of COX-2 was inhibited in a dose dependent manner when the compound (ASAP-calm) was treated.

Example  5: allergy  relation Eot Thaksin -1 secretion reduction effect measurement

Human normal fibroblast (HNF) was inoculated into a 24-well microplate containing DMEM medium to a concentration of about 2 x 10 ⁵ cells per well and cultured in a 5% CO ₂ incubator at 37 ° C for 24 hours Lt; / RTI > Then, 50 ng / ml of interleukin-4 (R & D, Minneapolis MN, IL-4) was treated with the compound (ASAP-calm) at concentrations of 1, 10 and 50 ppm The cell culture medium was collected. The degree of secretion of eotaxin-1 was then measured using an ELISA kit. As the control (-), a composite compound (ASAP-calm) and human fibroblasts which did not induce the secretion of eotaxin-1 were used. The results are shown in Fig.

As a result, it was shown that the secretion of eotaxin-1 induced by interleukin-4 was decreased by the complex compound (ASAP-calm). As the concentration of the compound (ASAP-calm) And the concentration-dependent tendency.

Example  6: Safety test of complex compounds against human skin

Since it was found that there was no cytotoxicity in the cytotoxicity test of Example 3, a skin safety verification experiment was conducted to confirm whether or not it was safe for human skin. As a test method, a normal cumulative skin irritation test was performed.

6-1. Preparation of external skin preparation containing complex compound (ASAP-calm)

In order to confirm that the complex compound (ASAP-calm) is safe for human skin, a skin external preparation containing a complex compound (ASAP-calm) was prepared by the ingredients and contents shown in Table 7 below, and skin stability and clinical efficacy test were conducted Respectively. First, purified water, glycerin, and butylene glycol were mixed and dissolved at 70 캜 (water part), and the other components except for the above three components and trimethanolamine were dissolved at 70 캜 (oil part). The oil part was added to a water part and stirred with a homomixer (Tokushu Kika, Japan) for primary emulsification, and trimethanolamine was finally added thereto. Next, the bubbles generated in the mixed solution were removed, and the mixture was cooled to room temperature to prepare an external preparation for skin.

ingredient content (%) Control group Test group 1 Test group 2 Purified water 72 72 72 glycerin 8.0 8.0 8.0 Butylene glycol 4.0 4.0 4.0 Complex compound (ASAP-calm) 0 0.1 1.0 Caprylic Capri Triglyceride 8.0 8.0 8.0 Squalane 5.0 5.0 5.0 Cetearyl glucide 1.5 1.5 1.5 Sorbitan stearate 0.4 0.4 0.4 Cetearyl alcohol 1.0 1.0 1.0 Trimethanolamine 0.1 0.1 0.1 Gross weight 100 100 100

6-2. Experiment of cumulative stimulation of skin

A total of 9 times and 24 hours cumulative pelletization was performed on the upper arm of the body in 30 healthy adults, and the compound (ASAP-calm) stimulated skin Respectively. As a control, an external preparation for skin without squalene-based complex compound (ASAP-calm) was used.

A pin chamber (Finn chamber, Epitest Ltd, Finland) was used as a method of applying the coating, and 15 μl of each of the external preparations was dropped into the chamber. The degree of the skin reaction on each occasion was scored using Equation 1 below, and the results are shown in Table 8 below.

[Experimental Equation 1]

(Number of total subjects × maximum score (4 points))} × 100] / number of tests (9 times)

At this time, a score of ± is given as 1, a score of 2 is given as +, and a score of 4 is given as ++, and a safe composition is judged when the average degree of reaction is less than 3.

Test substance Number of subjects who responded (persons) Average
Reactivity 1 week 2 weeks 3 weeks Primary Secondary Third Fourth 5th 6th Seventh 8th car 9th ± / + / ++ ± / + / ++ ± / + / ++ ± / + / ++ ± / + / ++ ± / + / ++ ± / + / ++ ± / + / ++ ± / + / ++ Control group - / - / - - / - / - - / - / - - / - / - - / - / - - / - / - - / - / - - / - / - - / - / - 0.00 Test group 1 - / - / - - / - / - - / - / - - / - / - - / - / - - / - / - - / - / - - / - / - - / - / - 0.00 Test group 2 - / - / - - / - / - - / - / - - / - / - - / - / - - / - / - - / - / - - / - / - - / - / - 0.00 Subject 30 30 30 30 30 30 30 30 30

As a result of the experiment, there was no reaction in 0, 0, 0 persons in the control group, the test group 1 and the test group 2, respectively. As a result of calculation according to the above formula, the average degree of reactivity of the control group and the test groups 1 and 2 was 0.00 or less, all 3 or less.

Example  7: Skin soothing effect  exam

After skin irritation was caused by SDS (0.05%) to damage skin tissue, skin external preparation (control group, test group 1, test group 2) prepared by the ingredients and contents of Table 7 was applied to the upper part of the skin, Was improved. In this experiment, 20 healthy adults were included.

A pin chamber (Finn chamber, Epitest Ltd, Finland) was used as the deposition method. 15 占 퐇 of each of the above-mentioned external preparations for skin was dropped into the chamber, followed by applying for 24 hours. After 24 hours, the amount of erythema induced by SDS was mitigated by each treatment substance using a Chroma meter CR-300 (Konica Minolta, Japan), and the degree of transdermal water loss (TEWL) was measured by TEWA Tewameter TM210, Germany). The cooling sensation was evaluated according to Table 9 below. The results are shown in Fig. 6 and Table 10.

+++ I felt a very strong cooling sensation. ++ I felt a strong sense of coolness. + I felt a refreshing feeling. - I do not feel a refreshing feeling.

Average response (refreshing feeling) Control group - Test group 1 ++ Test group 2 +++

As shown in FIG. 3, skin irritation induced skin irritation when treated with SDS, and excellent erythema reduction was observed when the compound (ASAP-calm) was treated. Furthermore, when the degree of transdermal water loss was measured, the transdermal water loss increased by the SDS treatment was remarkably alleviated by the compound (ASAP-calm).

On the other hand, as a result of the sensory test on the skin-cooling sensation under the respective conditions, it was confirmed that the frequency of responding to the refreshing feeling was significantly higher in the test group to which the compound (ASAP-calm) was applied.

Formulation example  One : Cosmetic  Formulation

1-1. Production of flexible lotion

As shown in the following Table 11, a lotion lotion containing a complex compound (ASAP-calm) as an active ingredient was prepared by a conventional method.

ingredient Content (% by weight) Complex compound (ASAP-calm) 0.1 glycerin 3.0 Butylene glycol 2.0 Propylene glycol 2.0 Carboxyvinyl polymer 0.1 ethanol 10.0 Triethanolamine 0.1 Preservatives, micronutrients, trace fragrances and trace water Balance sum 100.0

1-2. Manufacture of nutrition lotion

As shown in Table 12 below, a nutritional lotion containing a complex compound (ASAP-calm) as an active ingredient was prepared according to a conventional method.

ingredient Content (% by weight) Complex compound (ASAP-calm) 0.1 Wax 4.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.5 Liquid paraffin 5.0 Squalane 5.0 Caprylic / capric triglyceride 5.0 glycerin 3.0 Butylene glycol 3.0 Propylene glycol 3.0 Carboxyvinyl polymer 0.1 Triethanolamine 0.2 Preservatives, micronutrients, trace fragrances and trace water Balance sum 100.0

1-3. Manufacture of nutrition cream

A nutritional cream containing the complex compound (ASAP-calm) as an active ingredient was prepared according to a conventional method as shown in Table 13 below.

ingredient Content (% by weight) Complex compound (ASAP-calm) 0.1 Wax 10.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.5 Liquid paraffin 10.0 Squalane 5.0 Caprylic / capric triglyceride 5.0 glycerin 5.0 Butylene glycol 3.0 Propylene glycol 3.0 Triethanolamine 0.2 Preservatives, micronutrients, trace fragrances and trace water Balance sum 100.0

1-4. Manufacture of massage cream

A massage cream containing the compound (ASAP-calm) water as an active ingredient was prepared according to a conventional method as shown in Table 14 below.

ingredient Content (% by weight) Complex compound (ASAP-calm) 0.1 Wax 10.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.8 Liquid paraffin 40.0 Squalane 5.0 Caprylic / capric triglyceride 4.0 glycerin 5.0 Butylene glycol 3.0 Propylene glycol 3.0 Triethanolamine 0.2 Preservatives, micronutrients, trace fragrances and trace water Balance sum 100.0

1-5. Manufacture of packs

A pack containing the complex compound (ASAP-calm) as an active ingredient was prepared according to a conventional method as shown in Table 15 below.

ingredient Content (% by weight) Complex compound (ASAP-calm) 0.1 Polyvinyl alcohol 13.0 Sodium carboxymethylcellulose 0.2 Allantoin 0.1 ethanol 5.0 Nonyl phenyl ether 0.3 Preservatives, micronutrients, trace fragrances and trace water Balance sum 100.0

Claims (6)

A cosmetic composition for skin anti-irritation comprising an aloe extract, a Shin extract and a peppermint extract as an active ingredient.
The cosmetic composition for skin anti-irritation according to claim 1, wherein the aloe extract, Shin extract and peppermint extract are contained in a weight ratio of 0.9-1.1: 0.9-1.1: 0.9-1.1.
The cosmetic composition for skin anti-irritation according to claim 1, wherein the aloe extract, Shin extract and peppermint extract are contained in a weight ratio of 1: 1: 1.
[2] The composition according to claim 1, wherein the anti-irritant of the skin is used for improving or treating an inflammatory skin disease, exhibiting a cold effect for reducing inflammation and pain and a moisturizing effect for improving skin barrier, Or treating the skin.
The cosmetic composition for skin anti-irritation according to claim 1, wherein the cosmetic composition for skin anti-irritation is 0.0001 to 20% by weight based on the total weight of the composition.
The cosmetic composition of claim 1, wherein the cosmetic composition is a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray Lt; RTI ID = 0.0 > 1, < / RTI >

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