KR101664878B1 - Cosmetic composition containing fractions of allium hookeri as active ingredient - Google Patents

Abstract

Three of the present invention (Allium hookeri fractions have an effect of inhibiting MMP-1 production, enhancing collagen synthesis, inhibiting melanin production, and also improving skin wrinkle, skin whitening effect, skin elasticity improvement effect, skin aging prevention effect, antioxidative effect, skin barrier enhancement Effect, skin moisturizing effect, and preservative effect.

Description

FIELD OF THE INVENTION The present invention relates to a cosmetic composition containing a triple fraction as an active ingredient,

The present invention relates to a cosmetic composition containing a triangular fraction as an active ingredient.

A variety of physical and chemical changes occur in human skin during the aging process. The cause of this phenomenon is divided into intrinsic aging and photo-aging. Free radicals can be caused by activation of ultraviolet rays, stress, disease state, environmental factors, wounds, and aging. When such state is deepened, the antioxidant defense network existing in the living body is destroyed, And aging.

More specifically, lipids, proteins, polysaccharides and nucleic acids, which are major components of the skin, are oxidized to destroy skin cells and tissues, resulting in skin aging.

In particular, the oxidation of proteins can cause collagen (collagen), hyaluronic acid, elastin, proteoglycan, fibronectin, and the like that are severely hyperinflammatory reaction and skin elasticity If this gets worse, DNA mutations can lead to mutations, cancer, and immune deficiency.

Therefore, it is necessary to protect the cell membrane by destroying the free radicals mediated by free radicals, ultraviolet rays, and inflammation that occur during the metabolism of the body, and to regenerate already damaged cells by active metabolism, Can quickly recover and maintain healthy skin.

Aging involves not only these free radicals, but also enzyme called matrix metalloproteinase (MMP), a collagenase. That is, synthesis and degradation of extracellular matrix such as collagen in vivo are appropriately controlled, but collagen synthesis is reduced as aging progresses, and expression of matrix metalloproteinase (MMP), an enzyme that degrades collagen, is promoted, And the wrinkles are formed. These degrading enzymes are also activated by ultraviolet irradiation.

Therefore, there is a demand for development of a substance capable of modulating MMP expression induced in the cell or inhibiting its activity. Until now, the raw materials used as cosmetic materials mostly inhibited only the enzyme activity. Therefore, we tried to control the amount and activity of MMP expression induced by intracellular natural aging and photoaging.

On the other hand, skin changes due to pigment deposition occur. The factors that determine skin color are basically the partial or total pigmentation such as stain, freckle and tanning due to ultraviolet exposure, and the distribution of acne and scars and keratin Blood circulation, stress, and health. The most important factor among these factors is pigmentation.

Melanin, melanoid, carotene and hemoglobin are the most important factors affecting skin color. Among them, melanin is the most important factor affecting the biosynthesis of melanin.

Melanin absorbs or scatters ultraviolet light and plays a major role in preventing damage to the skin from ultraviolet rays. It does not have a specific maximum absorption wavelength and absorbs light in all areas. In addition, it has excellent ability to remove reactive oxygen species, sometimes melanin itself generates active oxygen, and other substances are reduced or oxidized by catechol or quinone in the melanin structure, and melanin itself shows free radical properties Pray.

The biosynthesis of melanin begins with the conversion of tyrosine, an amino acid, into melanosomes of melanocyte-forming melanosomes by tyrosinase and conversion to dihydroxy phenylalanine, followed by a series of oxidation processes to form pheomelanin, eumelanin.

This biosynthesis process is carried out in a special form of melanocyte in the brown intracellular organelle. The melanomas, including melanin granules, migrate from the nucleus to the tip of the dendritic process, and migrate into the cytoplasm by phagocytosis of keratinocytes. It accumulates around the nucleus.

The synthesis of melanin, the number of melanosomes, and the migration to keratinocytes in the periphery are partially affected by hormones and ultraviolet rays, and are primarily affected by genetics. In addition, it is known that interleukins, prostaglandins, histamines, and the like are involved in the expression of tyrosinase and cytokine, which is an intracellular regulator involved in the synthesis and transmission of melanin, and metal ions such as copper, zinc and iron.

(Japanese Patent Laid-open No. 4-9320), hydroquinone (Japanese Patent Laid-Open No. 192062), kojic acid (Japanese Patent Laid-open No. 56-7710), arbutin Activity, and is used as a whitening cosmetic. However, since it is low in stability in cosmetic formulations, its use is restricted due to decomposition and coloring, generation of odor, efficacy at a living body level, unclear effect and safety problem. These substances have proved their tyrosinase inhibitory effect, but their effect is lower in experiments similar to actual living body level. Therefore, it is required to evaluate the inhibitory effect by the melanoma cell culture similar to the biological level. Hydroquinone is also defined as a carcinogenic substance and its use in cosmetics is prohibited or restricted. Kojic acid and ascorbic acid are very unstable substances. When a cosmetic containing a small amount of the above components is kept at room temperature for several weeks, browning occurs.

In order to solve these various problems, recently, many cosmetics using natural products have been developed to reduce skin irritation caused by various chemical substances and the like. Natural materials are not only less harmful to the skin, but also have increased their value as a raw material for cosmetics due to the recent popularity of cosmetics using natural materials.

The present inventors have studied the applicability of cosmetics to various natural products that have not been known so far. As a result, the present inventors have found out that the lily plants such as Sanji (Sammi, Jumit and Root) , Collagen synthesis promotion effect, melanin production inhibitory effect, skin wrinkle improvement effect, skin whitening effect, skin elasticity improvement effect, skin moisturizing effect, preservative effect and the like, . Also,

It is still another object of the present invention to provide a cosmetic method using a cosmetic composition containing a triangular fraction.

The present invention relates to a process for the preparation of Allium hookeri fraction as an active ingredient.

Preferably, the cosmetic composition is for improving skin wrinkles.

Preferably, the cosmetic composition is for skin whitening.

Preferably, the cosmetic composition is for improving skin elasticity.

Preferably, the cosmetic composition is for preventing skin aging.

Preferably, the cosmetic composition has an antioxidative effect.

Preferably, the cosmetic composition has a skin barrier strengthening effect.

Preferably, the cosmetic composition is for skin moisturizing.

Preferably, the cosmetic composition has a preservative effect.

Preferably, the trifoliate fraction is contained in an amount of 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition.

Preferably, the trifoliate fraction is obtained by sequentially fractionating an extract obtained by extracting the trifoliate with an extraction solvent with N-hexane, methylene chloride, ethyl acetate and water.

Three of the present invention (Allium hookeri fractions have an effect of inhibiting MMP-1 production, enhancing collagen synthesis, inhibiting melanin production, and also improving skin wrinkle, skin whitening effect, skin elasticity improvement effect, skin aging prevention effect, antioxidative effect, skin barrier enhancement Skin moisturizing effect, and preservative effect (antibacterial effect).

Three are used as an active ingredient of the cosmetic composition of the present invention (Sammi holding, jyumit, roots and leek), Allium is in English representation is the scientific name It is labeled as hookeri . It is a perennial primrose of lily and wing. It is similar to the orange-yellow wave around 8-9. It is native to high altitudes of more than 1,400 meters in the Himalayas and is distributed in India, China, Bhutan, Sri Lanka and Myanmar. It is called "三 菜" because it has sweetness, bitter taste, and pungent taste. It is also called "玄 菜" because it is called "ginseng". The leaves and roots of the trichomes are supplemented with essential amino acids valine, isoleucine, methionine, threonine, lysine, phenylalanine, tryptophan, histidine, etc. And contains high levels of vitamin A, vitamin C, nitrogen, phosphoric acid, iron, manganese, zinc and sulfur. Particularly, the sulfur content of the tea is 3.28 mg per 100 g, which is 6 times higher than that of garlic, and in particular, it is known that 600 mg of dietary sulfur is contained per 100 g of the Korean liquor, They are not affected by wild animals even if they do not have a fence in the field.

In the present invention, the trichome fraction is obtained from various organs or parts (for example, leaves, flowers, roots, stems, etc.) of the trichomes, preferably a fraction obtained from the outposts or roots, and most preferably, Lt; / RTI >

In the present invention, the step (step 1) of obtaining a crude extract by extracting the triplex with an extraction solvent and the step (step 2) of fractionating the crude extract are obtained. Specifically, the step of fractionating the crude extract comprises sequentially fractionating the crude extract with n-hexane, methylene chloride, ethyl acetate and water (step 2). The step of obtaining crude extract (step 1) is extracted with an extraction solvent.

The extract can be extracted as an extraction solvent using water, an organic solvent or a mixed solvent thereof. The organic solvent is any one selected from the group consisting of C1 to C4 alcohols, ethyl acetate, methylene chloride and chloroform, or a mixed solvent thereof. Preferably, the organic solvent is extracted with a mixed solvent of water and a C1 to C4 alcohol, Preferably 60 to 100% ethanol. The temperature at the time of extraction is 30 占 폚 to 150 占 폚, preferably 50 占 폚 to 100 占 폚. The time for extraction is 30 minutes to 48 hours, preferably 1 hour to 12 hours. Extraction methods include ultrasonic extraction, supercritical extraction, subcritical extraction, hot water extraction, room temperature extraction or reflux extraction. Preferably room temperature extraction or reflux extraction, and more preferably, the reflux extraction is repeated 1 to 20 times, preferably 2 to 10 times. It will be apparent to those skilled in the art that the triple extract of the present invention can be obtained not only by the above-mentioned extraction solvent but also by using other extraction solvent, which exhibits substantially the same effect.

In addition, the tubular extract of the present invention includes not only the extract obtained by the above-described extraction solvent, but also the extract obtained through ordinary purification and fermentation processes. For example, decompression by carbon dioxide, extraction by supercritical extraction with high temperature, extraction by extraction using ultrasound, separation by ultrafiltration membrane with constant molecular weight cutoff value, various chromatography (size, charge, hydrophobic or affinity And the active fraction obtained through various purification and extraction methods, such as an extract obtained by fermentation products using natural or various microorganisms, are also included in the extract of the present invention.

The step of fractionating the crude extract (step 2) can be carried out in the same manner as in step 1 except that the crude extract obtained in the step of obtaining the crude extract (step 1) is neutralized with N-hexane, methylene chloride, ethyl acetate, And then sequentially fractionating the fractions.

The crude extract is redissolved in water, and the same amount of normal hexane as water is added. The mixture is fractioned 2 to 5 times. The normal hexane layer is separated and concentrated under reduced pressure to prepare a normal hexane fraction. Next, after removal of n-hexane, the same amount of methylene chloride is added to the remaining water layer, and the mixture is fractionated 2 to 5 times. Methylene chloride is separated and concentrated under reduced pressure to prepare methylene chloride fraction. After removing the methylene chloride, the same amount of ethyl acetate was added to the remaining water layer, which was then fractionated 2 to 5 times. Ethyl acetate was removed, and the filtrate was concentrated under reduced pressure to prepare ethyl acetate fraction. The remaining water layer is lyophilized to produce a water fraction.

According to the present invention, the trifoliate fraction is contained in an amount of 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition, and more preferably, the trifoliate fraction is contained in an amount of 0.01 to 10% by weight based on the total weight of the cosmetic composition do. When the content of the fraction is less than 0.0001% by weight, the skin improvement effect is not exhibited. When the content is more than 30.0% by weight, the skin improvement effect on the content increase is insignificant, It is not even.

The ingredients contained in the cosmetic composition of the present invention may contain, as an active ingredient, the ingredients commonly used in cosmetic compositions in addition to the above-mentioned effective ingredients, and may contain conventional ingredients such as antioxidants, stabilizers, solubilizers, vitamins, Supplements, and carriers. The cosmetic composition of the present invention can be prepared into any of the formulations conventionally produced in the art and can be used in the form of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, , Oil, powdered foundation, emulsion foundation, wax foundation, pack, massage cream and spray, but is not limited thereto. More specifically, it can be manufactured in the form of a soft 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 present invention is a paste, a cream or a gel, an animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide . When the formulation of the present invention 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 sorbitan fatty acid esters. In the case where the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as 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 tracant, etc. may be used. When the formulation of the present invention 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, / Propane or dimethyl ether. When the formulation of the present invention is a surfactant-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide ether Alkylamido betaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester.

When the cosmetic composition of the present invention is a soap, a surfactant-containing cleansing formulation or a surfactant-free cleansing formulation, it may be applied to the skin and then wiped off, washed or rinsed with water. As a specific example, the soap is liquid soap, powdered soap, solid soap and oil soap, and the surfactant-containing cleansing formulation is a cleansing foam, a cleansing water, a cleansing towel and a cleansing pack, , Cleansing lotion, cleansing water and cleansing gel, but is not limited thereto. On the other hand, the cosmetic process of the present invention refers to all cosmetic processes using the cosmetic composition of the present invention. That is, all methods known in the art using a cosmetic composition belong to the cosmetic method of the present invention. The cosmetic composition of the present invention may be used alone or in combination, or may be used in combination with other cosmetic compositions other than the present invention. The cosmetic composition according to the present invention may be used according to a conventional method of use, and may be used in a number of times depending on the skin condition or taste of the user. When the cosmetic composition comprising the tubular extract of the present invention is used, anti-inflammation, antibacterial, antioxidant, moisturizing, skin barrier strengthening effect can be obtained. Hereinafter, the present invention will be described in more detail with reference to Examples. These embodiments are only for describing the present invention more specifically, and the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention

The fractions obtained by this method are effective for inhibiting MMP-1 production, enhancing collagen synthesis, inhibiting melanin formation, improving skin wrinkles, skin whitening effect, skin elasticity improvement effect, skin aging prevention effect, Effect, and preservative effect (antibacterial effect).

[ Manufacturing example  1 to 4] Triptych Fraction  Produce

200g of the shrubs of the shrubs were shaken 3 times for 5 hours with 70% (v / v) ethanol aqueous solution, cooled, and filtered through Whatman # 4 filter paper to obtain an extract. Filtered and the filtrate was concentrated under reduced pressure and lyophilized to obtain 41.2 g of an ethanol extract. A systematic fractionation using an organic solvent was performed to fractionate the ethanol extract according to the polarity. Specifically, 20 g of the ethanol extract was re-dissolved in 200 mL of water, and an equal amount of n-hexane was added thereto. The hexane layer was separated to obtain a hexane fraction (Preparation Example 1). To the water layer from which the hexane fraction had been removed, methylene chloride was added in the same amount, and the methylene chloride layer was separated to obtain a methylene chloride fraction (Production Example 2). To the water layer from which the methylene chloride fraction had been removed, ethyl acetate was added in the same amount, and the ethyl acetate layer was separated to obtain an ethyl acetate fraction (Production Example 3). The ethyl acetate fraction was removed and the remaining water layer was lyophilized to give a water fraction (Preparation Example 4). The fractionation was carried out at room temperature and each fraction was allowed to stand for at least 3 hours. Each fraction was repeated twice. The fractions except for the water layer were filtered, concentrated under reduced pressure, and dried.

Experimental Example  One : Triptych Fraction MMP -1 production inhibitory effect

In Experimental Example 1, the effect of inhibiting the production of collagenase, MMP-1, of the triplex fractions obtained in Production Examples 1 to 4 was measured.

Human normal skin cells were inoculated into a 48-well microplate (Nunc, Denmark) at a concentration of 1 × 10 ⁶ cells per well and cultured in DMEM medium (Sigma, USA) and 37 ° C For 24 hours. Then, the cells were further cultured in serum-free DMEM medium supplemented with the triplicate fractions of Preparation Examples 1 to 4 and in serum-free DMEM medium containing no trivalent fraction for 48 hours.

After the incubation, the supernatant of each well was collected and the amount (ng / ml) of the newly synthesized MMP-1 was measured using an MMP-1 assay kit (Amersham, USA) The inhibition rate (%) was calculated and the results are shown in Table 1.

[Equation 1]

Name of sample Treatment concentration Inhibition rate of MMP-1 production (%) Production Example 1 Production Example 2 Production Example 3 Production Example 4 Trifoliate fraction 50 ppm 37.92 38.89 40.05 25.82 100ppm 49.50 56.14 59.48 35.75 500ppm 69.15 79.53 81.25 59.15 1000ppm 80.20 90.21 92.38 73.90 TGF-beta (200 nM) 76.80

As shown in the results of Table 1, it was confirmed that the triple fraction of the present invention inhibits the production of MMP-1 (inhibits MMP-1 activity) in a concentration-dependent manner, and is particularly excellent in Production Examples 2 and 3. Even considering the difference between the concentration of TGF-beta (200 nM), which is known to inhibit MMP-1 production, and the concentration (500 ppm) of the trifoliate fraction of the present invention, The inhibition of MMP-1 indicates that the triple fraction of the present invention has an inhibitory effect on MMP-1 production.

Experimental Example  2 : Triptych Fraction  Enhancement of collagen synthesis

Human normal epithelial cells, fibroblasts, were inoculated into 48-well microplates at a density of 1 x 10 ⁶ cells per well and cultured in DMEM medium for 24 hours. Serum-free DMEM medium supplemented with the fractions prepared in Preparation Examples 1 to 4 and the control group were further cultured for 48 hours in serum-free DMEM medium containing no trivalent fraction. After the incubation, the supernatant of each well was collected, and the amount of procollagen type I C-peptide (PICP) was measured using a collagen kit (Takara, Japan) and converted into ng / Respectively. Collagen biosynthesis increase rate (%) was calculated according to the following formula (2), and the results are shown in Table 2.

&Quot; (2) "

Name of sample Treatment concentration Increase in collagen biosynthesis (%) Production Example 1 Production Example 2 Production Example 3 Production Example 4 Trifoliate fraction 5 ppm 15.3 20.1 25.5 11.5 10ppm 28.4 45.1 50.1 27.7 25 ppm 82.3 97.5 100.5 41.5 50 ppm 121.5 162.4 170.5 94.8 TGF-beta (200 nM) 165.5

According to Table 2, it was confirmed that the collagen biosynthesis rate of the trifoliate fraction prepared in Production Examples 1 to 4 of the present invention was increased in a concentration-dependent manner. In particular, it was confirmed that the trihydrate fraction produced by Production Examples 2 and 3 had excellent effects.

Experimental Example  3: Triptych Fraction  Effect of B16F1 melanin-forming cells on melanin formation

In Experimental Example 3, in order to confirm the whitening effect of the triplet fractions obtained in Production Examples 1 to 4, the degree of inhibition of melanin formation on B16F1 melanocyte-forming cells was examined to determine the whitening effect. The B16F1 melanin-forming cell used in Experimental Example 3 is a cell strain derived from a mouse, and is a cell that secretes a melanin pigment called melanin.

During the artificial culture of these cells, samples were treated to compare the degree of reduction of melanin pigment. The B16F1 melanin-forming cells used in this experiment were purchased from the American Type Culture Collection (ATCC).

Melanin biosynthesis inhibitory effect of B16F1 melanin-forming cells was measured as follows.

B16F1 melanogenesis cells were seeded at a concentration of 2 × 10 ⁵ per 6 wells on a 6-well plate, and the cells were treated with the triplicate fraction according to Preparations 1 to 4 at a concentration that did not cause toxicity, followed by incubation for 72 hours. After incubation for 72 hours, the cells were detached with trypsin-EDTA, the number of cells was measured, and the cells were recovered by centrifugation. Quantification of intracellular melanin was carried out with a slight modification of the method of Lotan (Cancer Res., 40: 3345-3350, 1980). Cell pellet was washed once with PBS, and 1 ml of homogenization buffer solution (50 mM sodium phosphate, pH 6.8, 1% Triton X-100, 2 mM PMSF) was added and vortexed for 5 minutes to disrupt the cells. After centrifugation (3,000 rpm, 10 min), 1N NaOH (10% DMSO) was added to the cell filtrate to dissolve the extracted melanin, and the absorbance of melanin was measured at 405 nm with a microplate reader. The inhibition rate (%) of melanin formation was measured. Melanin formation inhibition rate (%) of B16F1 melanin-forming cells was calculated by the following equation (3), and the results are shown in Table 3. [

&Quot; (3) "

Melanin formation inhibition rate (%) = [(A-B) / A] X100

A: Amount of melanin in wells to which no sample was added

B: Amount of melanin in the well to which the sample was added

Name of sample Treatment concentration (%) Melanin formation inhibitory effect (%) Production Example 1 Production Example 2 Production Example 3 Production Example 4 Trifoliate fraction 0.001 12.8 19.27 23.11 11.5 0.005 28.4 37.89 42.16 27.7 0.01 41.5 53.72 56.42 40.5 0.05 51.2 74.11 77.42 48.4 Arbutin 200ppm 68.25

As can be seen from the results of Table 3, it was found that the purified triangular fraction prepared according to Production Examples 1 to 4 of the present invention significantly inhibited melanogenesis in B16F1 melanin-forming cells.

Experimental Example  4 : Triptych Fraction  Antioxidative effect

The antioxidative effect (free radical scavenging ratio) was measured by the DPPH method in order to measure the antioxidative effect of the trivalent fractions obtained in Production Examples 1 to 4. The DPPH method measures the antioxidative effect by reducing power using a free radical called DPPH (2,2-Di (4-tert-octylphenyl) -1-picrylhydrazyl) free radical. The degree of decrease in absorbance of DPPH is reduced by the test substance (elderberry extract) to the absorbance of the blank test solution and the free radical scavenging ratio (%) is measured at a wavelength of 560 nm. As a reagent used, 61.88 mg of a 0.1 mM solution of 2,2-Di (4-tert-octylphenyl) -1-picrylhydrazyl free radical (Aldrich Chem. Co., MW = 618.76) was dissolved in methanol to make 100 ml.

As a measuring method

(1) Add 0.15 ml of the sample solution to 0.15 ml of 0.1 mM DPPH solution in a 96-well plate, stir rapidly, and start culturing at 25 ° C for 10 minutes.

② Measure the absorbance St at 560 nm thereafter.

(3) The blank of the sample solution is measured by the same procedure as that for measuring the absorbance St except that methanol is used instead of the 0.1 mM DPPH solution.

(4) The absorbance Bt is measured by operating in the same manner as in the measurement of the absorbance St except that distilled water is used instead of the sample solution.

(5) Blank of the blank test is carried out in the same manner as in the measurement of the absorbance St except that methanol is used instead of the 0.1 mM DPPH solution and distilled water is used instead of the sample solution, and the absorbance Bo is measured.

The results of the free radical scavenging rate (%) were calculated according to Equation (3), and the results are shown in Table 5 below.

&Quot; (4) "

Free radical scavenging rate (%) = [1 - {(St-So) / (Bt-Bo)}] x100

St: absorbance at 514 nm after free radical scavenging of the sample solution

Bt: Absorbance at 514 nm after free radical scavenging of blank test solution

So: the absorbance at 514 nm before the reaction in the absence of free radicals in the sample solution

Bo: absorbance at 514 nm before the reaction in the absence of the free radical of the blank test solution

Name of sample SC ₅₀ (%) Production Example 1 0.08 Production Example 2 0.01 Production Example 3 0.015 Production Example 4 1.0

SC ₅₀ , which is a concentration of a sample required for 50% DPPH free radical scavenging, was confirmed. The results are shown in Production Examples 1 to 4 and especially in Production Examples 2 and 3.

Formulation Example  1 to 4: Triptych The fraction  Containing Cosmetics  Preparation of composition

The cosmetic compositions containing the trifoliate fractions were each prepared so as to contain the trifoliate fractions of Production Examples 1 to 4. For comparison of the efficacy, a cosmetic composition containing glycerin and 1,3-butylene glycol as moisturizing agents instead of the trifoliate fraction (Comparative Formulation Example 4) containing the cosmetic composition (Comparative Formulation Example 3) and ceramides (Comparative Formulation Example 4) containing no fractions or moisturizing agents are shown in Table 5 below Respectively.

division ingredient Formulation Example 1 Formulation Example 2 Formulation Example 3 Formulation Example 4 compare
Formulation Example 1 compare
Formulation Example 2 compare
Formulation Example 3 compare
Formulation Example 4 A Production Example 1 2.0 - - - - - - - Production Example 2 - 2.0 - - - - - - Production Example 3 - - 2.0 - - - - - Production Example 4 - - - 2.0 - - - - Adenosine - - - - - 3 - - glycerin - - - - 5 - - - 1,3-butylene glycol - - - - 5 - - - Ceramide - - - - - - - 0.5 EDTA-2Na 0.02 0.02 0.02 0.02 0.02 0.02 0.02 0.02 Purified water to 100 to 100 to 100 to 100 to 100 to 100 to 100 to 100 B Cetostearyl alcohol 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 Glyceryl stearate 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 Microcrystalline 0.7 0.7 0.7 0.7 0.7 0.7 0.7 0.7 Squalane 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 Liquid paraffin 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Trioctanoin 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 Polysorbate 1.2 1.2 1.2 1.2 1.2 1.2 1.2 1.2 Sorbitan stearate 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Tocopheryl acetate 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 Cyclomethicone 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 BHT 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 C Incense, preservative Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount

Experimental Example  5: Triptych Fraction  Wrinkle improvement effect

Clinical experiments on the effect of improving the skin wrinkles of the cosmetic compositions containing the triangular fraction of the present invention were conducted. Clinical trials were conducted through actual use tests of each cosmetic product.

That is, the clinical trial was conducted in the same manner as in Formulation Examples 1 to 4 containing the triple fraction of Production Examples 1 to 4 in Table 6 and Comparative Formulation Example 1 containing glycerin and 1,3-butylene glycol as moisturizing agents The cosmetic composition was used to confirm and compare the effect of improving the skin wrinkles caused by the use of cosmetics on humans.

120 female subjects (35-50 year-old female) were divided into experimental group (20 patients each), cosmetic composition (Comparative Formulation Example 1) containing glycerin and 1,3-butylene glycol as moisturizing agents, The triplet fractions were divided into 6 groups of the cream (Comparative Formulation Example 2) treatment group in which adenosine was replaced with 3% (v / v) and the experimental group of Formulation Examples 1 to 4 containing the triple fraction, The degree of wrinkle improvement was evaluated by visual evaluation after wrinkle improvement. The results of the wrinkle-improving effect based on this evaluation are shown in Table 7 below.

The effect of improving wrinkles after the completion of the experiment was evaluated by visual examination and instrument measurement (cutometer SEM 575, C + K Electronic Co., Germany) of a skilled physician before and after 3 months of use. The experimental results are shown in Table 6 below.

Wrinkle improvement effect Effective rate (%) Great slightly none Formulation Example 1 12 2 6 70.0 Formulation Example 2 14 4 2 90.0 Formulation Example 3 15 3 2 90.0 Formulation Example 4 9 6 7 65 Comparative Formulation Example 1 3 3 14 30.0 Comparative Formulation Example 2 14 2 3 85

As can be seen from the results of Table 6 above, the cosmetic compositions of Formulation Examples 1 to 4 containing the triangular fraction of the present invention showed a wrinkle-reducing effect. Particularly, Formulation Examples 2 and 3 are superior to Comparative Formulation Example 1 containing moisturizing agent Showed about 3.0 times higher wrinkle improvement effect.

In addition, similar results were obtained with the cosmetic preparation of Comparative Formulation Example 2 containing adenosine, which is known to have a wrinkle-reducing effect instead of the fructose-containing fraction, indicating that the fructose-containing fraction exhibits excellent wrinkle-improving effect.

Experimental Example  6: Measurement of skin elasticity improvement effect

In Experimental Example 6, to evaluate the skin elasticity improving effect of the cosmetic composition according to the formulation examples and the comparative formulations including the triangular fraction obtained in Production Examples 1 to 4, 75 female subjects with reduced elasticity in 30 to 40 years . 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, 15, For 12 weeks, twice a day, and then skin elasticity was measured after 12 weeks. The elasticity of the facial region was measured using a cutometer SEM575 (MPA 580, Courage + Khazaka GmbH, Germany), and the R2 (biological elasticity) indicating skin elasticity was measured before and after application . The results are shown in Table 8 below, and the results are the average of the degree of improvement for each group.

Experimental product Skin elasticity improvement (%) Formulation Example 1 15.7 Formulation Example 2 24.8 Formulation Example 3 25.4 Formulation Example 4 13.4 Comparative Formulation Example 1 4.7

As shown in Table 7, Formulations 1 to 4 containing the triangular fraction had significantly improved skin elasticity as compared to Comparative Formulation Example 1.

Experimental Example  7: Skin barrier strengthening effect TEWL )

TEWL (Trans-Epidermal Water Loss) is an index of moisture (excluding sweat) that evaporates through the skin and is used to assess the extent of damage to the stratum corneum and skin barrier function. A high level of TEWL indicates that the skin barrier is damaged. TEWL was measured at constant temperature (22 ° C) and humidity (47%), and the unit was g / h / m ² . The method of measurement was as follows: First, 30 times of tape-stripping was performed to cause damage to the skin barrier. TEWL levels were measured using a Tewameter TM210 (Courage + Khazaka, Germany) prior to application of the test product after causing skin barrier damage. Formulations 1 to 4 and Comparative Formulations 3 and 4 of Table 5 were applied to damaged skin for 2 days twice a day for 3 days and the change in TEWL value was measured to measure the recovery of transdermal water loss according to Equation 5 . The results are shown in Table 8.

&Quot; (5) "

Percutaneous water loss recovery (%) = (TEWL immediately after disturbance - TEWL at indicated time) / (TEWL immediately after disturbance - baseline TEWL) x100

Name of sample TEWL value TEWL Recovery (%) Before stripping After stripping  Three days after application Formulation Example 1 6.9 19.2 9.1 79.1 Formulation Example 2 7.5 18.2 7.9 96.3 Formulation Example 3 7.2 19.6 7.5 97.6 Formulation Example 4 7.0 19.5 8.5 76.0 Comparative Formulation Example 3 7.0 19.7 15.8 30.7 Comparative Formulation Example 4 6.8 18.9 7.1 97.5

As shown in Table 8, Formulation Examples 1 to 4 containing the trivalent fractions, Comparative Formulation Example 4 containing the ceramide, and Comparative Formulation Example 3 containing no active ingredient were applied to the skin, and the TEWL was measured to confirm the skin barrier strengthening effect As a result, Formulation Example 2 and Formulation 3 containing methylene chloride and ethyl acetate fractions showed particularly excellent effects and showed similar effects to Comparative Formulation Example 4 containing ceramides.

Experimental Example  8 : Triptych Fraction  Moisturizing effect

The clinical moisturizing effect of the cosmetic composition of the above formulation example was measured as follows. A, B, C, and D groups were divided into seven groups (A, B, C, D, E, F) (Comparative Formulation Example 1) containing glycerin and 1,3-butylene glycol as a moisturizing agent instead of the triple fraction in the E group and a formulation containing no moisturizing agent in the F group Example 3) were applied to the face for 2 weeks each for 4 weeks. The moisturizing effect was evaluated by Corneomater CM820 (Corage + Khazaka, Germany) after every two weeks and after the start of the test. The results are shown in Table 9 below.

division Before use After 2 weeks of use After 4 weeks of use Formulation Example 1 29.15 44.15 51.12 Formulation Example 2 28.98 45.19 53.89 Formulation Example 3 30.81 37.18 54.19 Formulation Example 4 31.11 43.15 49.45 Comparative Formulation Example 1 30.3 42.1 50.9 Comparative Formulation Example 3 28.5 29.4 28.9

Experimental Example  9: Triptych  Preservative effect of extract

In order to examine the preservative effect of the tubular extracts, the cosmetic compositions of Formulation Example 5-12 containing the tubular extracts of Preparation Examples 1, 2 and 3 at various ratios in the composition of the following Table 10, Comparative Formulations containing purified water instead of the tubular extract The cosmetic composition of Comparative Formulation Example 6 containing methylparaben and imidazolidinyl urethane as a preservative instead of 5 cosmetic and triple extracts was prepared.

ingredient Formulation Example Comparative Formulation Example 5 6 7 8 9 10 11 12 5 6 Content (unit:% by weight) Trifoliate fraction
(Production Example 1) 0.5 1.0 - - - - - - - - Trifoliate fraction
(Production Example 2) - - 0.5 1.0 - - - - - - Trifoliate fraction
(Production Example 3) - - - - 0.5 1.0 - - - - Trifoliate fraction
(Production Example 4) - - - - - - 0.5 1.0 - - glycerin 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 Butylene glycol 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 Propylene glycol 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 EDTA-2Na 0.02 0.02 0.02 0.02 0.02 0.02 0.02 0.02 0.02 0.02 Polysorbate 60 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 Glyceryl stearate 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 Wax 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 Macadamia nut oil 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 Squalane 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Spices a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Methylparaben - - - - - - - - - 0.2 Imidazolidinyl urea - - - - - - - - - 0.2 Purified water To 100 To 100 To 100 To 100 To 100 To 100 To 100 To 100 To 100 To 100

The cosmetic effects of the formulation examples and comparative formulations of Table 10 were used to measure the preservative effect on bacteria and fungi.

First, in the case of bacteria, the cosmetic compositions of Formulation Examples 5 to 12 and the cosmetic compositions of Comparative Formulation Examples 5 and 6 were mixed with 20 to 30 g of Escherichia coli ; ATCC 8739), Pseudomonas aeruginosa (ATCC 9027) and Staphylococcus aureus (ATCC 6538) were added and mixed at an initial concentration of 10 ⁷ cfu / g per sample. 1 g of each cosmetic was taken at 1, 7, 14, 21 and 28 days intervals while culturing them in a thermostat at 30-32 ° C for 4 weeks, and the number of viable cells was measured. The measurement results are shown in Table 11 below.

Next, in the case of mold, Candida albicans ( Candida albicans ; ATCC 10231), Penicillium citrinum ; KCTC2123) and Aspergillus niger (ATCC16404) were added at a concentration of 10 ⁷ cfu / g per sample and cultured at 25 ° C in a thermostatic bath at intervals of 7 days, And the results are shown in Table 11 below.

Cosmetics Bacteria (cfu / g) mold* Initial number of bacteria Day 1 Day 7 Day 14 Day 21 Day 28 Formulation Example 5 1 x 10 ⁷ 600000 25000 5000 500 <100 - Formulation Example 6 1 x 10 ⁷ 310000 4000 500 <100 <100 - Formulation Example 7 1 x 10 ⁷ 650000 31000 8000 600 <100 - Formulation Example 8 1 x 10 ⁷ 360000 5000 650 200 <100 - Formulation Example 9 1 x 10 ⁷ 690000 38000 7000 950 <100 - Formulation Example 10 1 x 10 ⁷ 390000 6500 810 400 <100 - Formulation Example 11 1 x 10 ⁷ 710000 100000 80000 25000 1200 - Formulation Example 12 1 x 10 ⁷ 580000 13000 5000 900 <100 - Comparative Formulation Example 5 1 x 10 ⁷ 8500000 8000000 7400000 6300000 5900000 +++ Comparative Formulation Example 6 1 x 10 ⁷ 350000 7000 800 <100 <100 -

-: No spawning and mycelium spawning for 8 weeks and good

+: Molding on the wall or lid within 4 weeks

++: mending within 4 weeks and mold on part of the surface

+++: Mildew and mold on the entire surface within 4 weeks

As shown in Table 11, the cosmetic composition of Comparative Example 5, which did not use an antiseptic, showed molds on the surface and the entire surface within 4 weeks, and bacterial bacteria were remarkably increased even after 1 day. However, the cosmetic compositions of Formulation Examples 5 to 12 containing the triple extract of the present invention showed a concentration-dependent preservative effect on bacteria and fungi, and after about 28 days, 2.0% (w / v) The preservation force similar to or better than Comparative Formulation Example 6 in which the preservative methylparaben and imidazolidinyl urea were mixed was obtained and when it was judged according to the whole culture day that the preservative was used in an amount of 2.0% It was found that a far superior buoyant force was obtained as compared to Comparative Formulation Example 6 in which methylparaben, which is an imidazolidinyl urea, and imidazolidinyl urea were mixed. In particular, it was confirmed that the tubular extracts prepared in Preparation Examples 1 and 2 were more excellent in flotation power.

As shown in Table 11, it was confirmed that the moisturizing effect of Formulation Examples 1 to 4 containing the triacylglycerol fraction of the present invention was superior to Comparative Formulation Example 1 containing other moisturizing agents.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Claims (11)

A cosmetic composition for enhancing skin barrier comprising, as an active ingredient, a normal hexane fraction, a methylene chloride fraction or an ethyl acetate fraction of a tubular extract. A cosmetic composition for skin moisturizing, comprising a normal hexane fraction, a methylene chloride fraction or an ethyl acetate fraction of a tubular extract as an active ingredient. 3. The method according to claim 1 or 2,
The trifoliate fraction is a cosmetic composition which is a normal hexane fraction, a methylene chloride fraction or an ethyl acetate fraction obtained by sequentially fractionating a tubular extract with N-hexane, methylene chloride, ethyl acetate and water . 3. The method according to claim 1 or 2,
Wherein said trifoliate fraction is contained in an amount of 0.0001 to 30.0% by weight based on the total weight of said cosmetic composition. delete delete delete delete delete delete delete