CN108066170A - Dermatologic preparation composition containing ginsenoside Mc - Google Patents

Abstract

The present invention relates to a kind of compositions for containing ginsenoside Mc as active ingredient, the composition can not only provide anti-aging, improvement wrinkle of skin, whitening, moisturizing improvement, but also it is capable of providing anti-inflammatory, improvement acne and skin problem, and effect, convergence skin and the pore refining effect of allergic symptom, and be capable of providing skin color improvement, promote hair growth, improve white hair, anti-dandruff and anti-corrosion effect.

Description

Dermatologic preparation composition containing ginsenoside Mc

The application is Application No. 201480029499.7, the applying date to be on April 24th, 2014, entitled " contain The divisional application of the Chinese patent application of the Dermatologic preparation composition of ginsenoside Mc ".This application claims April 24 in 2013 The priority for the 10-2013-0045117 korean patent applications that day submits in Korean Intellectual Property Office.

Technical field

The present invention relates to a kind of composition, the composition contains ginsenoside Mc (ginsenoside Mc), so as to not Only be capable of providing it is anti-aging, improve wrinkle of skin, whitening, moisturizing improvement, but also be capable of providing it is anti-inflammatory, improve acne and The effect of skin problem and allergic symptom, convergence skin and pore refining effect, and it is capable of providing skin color improvement effect Fruit promotes hair growth, improves white hair, anti-dandruff and anti-corrosion effect.

Background technology

First defensive barrier of the skin as human body, have so that intracorporeal organ from temperature and humidity variation and it is ultraviolet The stimulations of external environments such as line, public hazards substance and protect the function of intracorporeal organ.With advancing age, skin will be due to a variety of Inherent, external factor and change.That is, in terms of the inherence, since the various hormone secretions for adjusting metabolism are reduced, And the function and cytoactive of immunocyte reduce, therefore the biology of the immune protein of needed by human body and organism constitutive protein Synthesis is reduced.For in terms of external, due to the destruction of ozone layer, the ultraviolet light content that earth's surface is reached from sunray increases Add, and as the in-depth of environmental pollution, free radical and active oxygen increase, the thickness reduction of skin, wrinkle is not only caused to increase, Elastic force reduces, and makes skin complexion dimmed, and problem (trouble) often occurs for skin, can also increase mole and freckle and old age Spot, and cause the various changes such as complexion is deteriorated, the colour of skin is dimmed.

In order to prevent these due in skin and external factor and occur skin condition variation, and maintain health skin Skin state, people make great efforts always by the way that the physiological activator obtained from existing various animals, plant, microorganism etc. is added Enforcement of going forward side by side is added in cosmetics to be used for improving skin condition.

Ginsenoside Mc be the natural goods form contained in ginseng ginsenoside (Rb1, Rg1, Rd, Rb2, Rc, Rf) by What digestive ferment and intestinal microbial were decomposed and generated.It has been reported that the active anticancer of ginsenoside Mc is strong and Korean granted is special The method for using ginsenoside Mc as anticancer preparation is recorded in profit the 164266th, but without reporting ginsenoside Mc is as such as whole skin condition improvement of the composition of active ingredient, promotion hair growth and improves white hair, anti-head The overall effect as skin preparations for extenal use such as scurf and anti-corrosion.

The content of the invention

Technical problems to be solved

In this regard, the inventors discovered that the ginsenoside Mc contained in ginseng can not only provide anti-aging, improvement skin wrinkle Line, whitening and moisturizing improvement, but also effect that is anti-inflammatory, improving acne, skin problem and allergic symptom is capable of providing, and Skin color improvement is capable of providing, promotes hair growth, improve white hair, anti-dandruff and anti-corrosion effect, so as to complete The present invention.

Therefore, it is an object of the present invention to provide a kind of Dermatologic preparation composition, the composition contains ginsenoside Mc, so as to show that the integrality of skin improves and promote hair growth, improvement white hair, anti-dandruff and anti-corrosion effect Fruit.

Technical solution

To achieve these goals, present invention offer is a kind of contains ginsenoside Mc as active ingredient for anti-aging Dermatologic preparation composition.

In addition, the present invention provides a kind of external preparation for skin for being used to improve wrinkle for containing ginsenoside Mc as active ingredient Agent composition.

In addition, the present invention provides a kind of skin preparations for extenal use group for moisturizing for containing ginsenoside Mc as active ingredient Close object.

In addition, the present invention provides a kind of external preparation for skin for being used to improve acne for containing ginsenoside Mc as active ingredient Agent composition.

In addition, the present invention provides a kind of skin for being used to improve color and the colour of skin for containing ginsenoside Mc as active ingredient Skin preparation composition for external use.

In addition, the present invention provides a kind of external preparation for skin for pore refining for containing ginsenoside Mc as active ingredient Agent composition.

In addition, the present invention provides a kind of skin for being used to improve allergic skin for containing ginsenoside Mc as active ingredient Skin preparation composition for external use.

In addition, the present invention provides a kind of ginsenoside Mc that contains is used for anti-inflammatory skin preparations for extenal use group as active ingredient Close object.

In addition, the present invention provides a kind of skin preparations for extenal use group for whitening for containing ginsenoside Mc as active ingredient Close object.

In addition, the present invention provides a kind of ginsenoside Mc that contains is used for trichogenous skin as active ingredient Preparation composition for external use.

In addition, the present invention provides a kind of external preparation for skin for being used to prevent white hair for containing ginsenoside Mc as active ingredient Agent composition.

In addition, the present invention provides a kind of external preparation for skin for anti-dandruff for containing ginsenoside Mc as active ingredient Agent composition.

In addition, the present invention provides a kind of natural antiseptic agent composition by ginsenoside Mc.

Advantageous effect

The composition of the present invention contains ginsenoside Mc, so as to provide anti-aging, improvement wrinkle of skin, U.S. In vain, moisturizing improvement, but also it is capable of providing anti-inflammatory, improvement acne and the effect of skin problem and allergic symptom, convergence Skin and pore refining effect, and be capable of providing skin color improvement, promote hair growth, improve white hair, anti-dandruff And anti-corrosion effect.

Specific embodiment

Dermatologic preparation composition according to the present invention contains ginsenoside Mc as active ingredient.

The ginsenoside Mc used in the present invention has the structure of following chemical formula 1：

[chemical formula 1]

The ginsenoside Mc of the present invention can be extracted from plant, can also be synthesized simultaneously by method as known in the art It uses, the ginsenoside Mc of mercantile-type sale can also be used.In addition, ginsenoside Mc can be obtained from ginseng extract. At this moment the species of the ginseng used is not particularly limited, and can use water ginseng, red ginseng, white ginseng, Tai Ji ginseng and tail ginseng etc..Also, The ginseng extract not only comprising extracted as ginseng, decoct carry obtained from leachate, also include to the part or whole of leachate Concentrate obtained from body is concentrated again or impregnating, decoction, the tablet dried the concentrate again and preparedFlowing extracts and the chemical substance that main efficacy results are played included in ginseng, but also include plant sheet Body, and the extract of the entire parts of ginsengs such as stem, root, leaf, flower and fruit can be used, it is not limited to a certain specific part Extract.In addition, the method for ginsenoside Mc is extracted from ginseng extract can use well known method.

Specifically, the ginsenoside Mc can be made by method as known in the art using water or organic solvent After standby ginseng extract, therefrom it is isolated.The organic solvent used in the present invention can be selected from ethyl alcohol, methanol, butanol, Ether, ethyl acetate, chloroform and the mixed solvent of these organic solvents and water, it is preferable to use 80% ethyl alcohol.At this moment, Extracting temperature Preferably 10~80 DEG C, and can extract 3~24 it is small when.If beyond the Extracting temperature and the scope of extraction time, Extraction efficiency can reduce or can occur the variation of ingredient.

Composition according to the present invention in terms of composition total weight, preferably comprises the ginsenoside of 0.001 to 50 weight % Mc.If this is because the content of the ginsenoside Mc is less than 0.001 weight %, the effect of being brought by the ingredient and effect Fruit is faint, if it exceeds 50 weight %, then can there are problems that in cutaneous safety or dosage form.

The composition of the present invention can be used as and be used for anti-aging Dermatologic preparation composition, in raising skin Effect in terms of elastic force and improvement wrinkle protrudes.

The composition of the present invention can be used as the Dermatologic preparation composition for moisturizing, can strengthen skin Barrier function, and it is capable of the differentiation of induced skin keratinocyte.Therefore, can effectively serve as preventing or improving because of epidermis Not xerodermia, allergic dermatitis, the skin preparations for extenal use for contacting atopic dermatitis or psoriasis etc. completely caused by differentiation Composition.

The composition of the present invention can be used as improving the Dermatologic preparation composition of acne, antibacterial effect It is excellent, it is especially excellent to the antibacterial effect of acne pathogenic bacteria, and antiphlogistic effects are provided.

The composition of the present invention can be used as improving the Dermatologic preparation composition of color and the colour of skin, by it When being used in skin, by expanding capillary, and promote blood circulation that nutritional ingredient smoothly is fed to skin, and inhibit Skin aging, therefore, the effect for improving color and the colour of skin are remarkable.

The composition of the present invention can be as pore refining, adjusting sebum and the skin preparations for extenal use for improving skin problem Composition uses, and when being used in skin, inhibits the sebum of excessive secretion, and passes through the elimination for promoting active oxygen and collagen The synthesis of albumen carrys out pore refining, and due to the reduction of inflammatory Cytokines Expression, the effect for inhibiting skin problem is remarkable.

The composition of the present invention can be used as improving the composition of allergic skin, not only pass through inhibition Induce the work of the protein decomposition enzyme active acceptor -2 (Proteinase-Activated Receptor-2, PAR-2) of itch Property, so as to provide excellent antipruritic effect, but also can be by reducing the secretion of interleukin (Interleukin-8, IL8) To provide anti-inflammatory effect.Therefore, the ginsenoside Mc of the invention can be used as stabilized susceptibility, irritation or mistake Quick property skin and scalp, and it is appropriate for improving or alleviating the active ingredient of the Dermatologic preparation composition of thermal sensation and inflammation Ground uses.

The composition of the present invention can be used as the composition for whitening, by the work for hindering tyrosinase Property, inhibit the generation of melanin, so as to provide excellent whitening effect.

The composition of the present invention can be used as and be used for trichogenous Dermatologic preparation composition, promote From hair cycle stand-down to the conversion in anagen hair cycle, so as to provide the growth including promoting hair, and promote new The generation of hair, and make the effect of existing hair health growth also provides and prevents and inhibit hair to show from what scalp came off As or hair is scattered or the effect of state that attenuates.

The composition of the present invention can be used as preventing the Dermatologic preparation composition of white hair, pass through raising The expression of transcription factor (MITF) in melanocyte carrys out activation of melanocyte, and promotes the synthesis of melanin, so as to provide The induction of advance preventing white hair, and promote to induce the effect of dark hair.

The composition of the present invention can be used as preventing the Dermatologic preparation composition of dandruff, by having Effect discharge is accumulated in the toxin on hair and scalp to purify scalp, and by inhibiting multiplication and the growth of dandruff bacterium and can Prevent scalp inflammation reaction, also, since the anti-oxidation efficacy of generation and the effect of inhibitory activity oxygen protrudes, therefore, it is possible to carry For calming and strengthening scalp, and strengthen the effect of intrinsic phylactic power defensive power.

The composition of the present invention can be used as natural antiseptic agent composition, since it is natural component, anti-corrosion It is also harmless while effect brilliance.

The composition of the present invention can the agent containing acceptable carrier or base material on cosmeceutical or Dermatology Type.It can be provided as the whole dosage forms for being suitble to locally use, for example, can be provided as solution, gel, solid, paste The anhydrous product of shape In water phase disperse oil phase and obtain lotion, suspension, microemulsion, Microcapsules, subparticle ball or ionic (liposome) and non-ionic folliculus dispersant form or frost, toner, wash Agent, powder, ointment, spray or the form for hiding flaw stick.And it is possible in the form of foam (foam) or further include pushing away for compression Form into the aerosol combination of agent uses.These compositions can be prepared according to method commonly used in the art.

In particular, the Dermatologic preparation composition of the present invention is used to prevent dandruff, is for hair growth or white for preventing During hair, can as the composition for scalp and hair and dosage form, dosage form are not particularly limited, such as can be turned to dosage form Hair oil, trichotrophy toner, scalp care agent, hair-care agent, shampoo, hair conditioner, hair lotion or hair of scalp Dual-purpose care agent etc..

In addition, composition according to the present invention can include fatty material, organic solvent, lytic agent, concentrating agents, gelling Agent, softening agent, antioxidant, suspending agent, stabilizer, foaming agent (foaming agent), aromatic, surfactant, water, Ionic or nonionic emulsifier, filler, chelating agent, complexing agent, preservative agent, vitamin, blocking agent, wetting agent, essential oil, Dyestuff, pigment, hydrophily or lipophile activating agent, lipid folliculus or any other ingredient for being usually used in cosmetics etc. are in cosmetics Common adjuvant in or Dermatology field.The adjuvant is with usually used in cosmeceutical or Dermatology field Amount import.

In addition, the composition of the present invention, in order to increase skin improvement effects, can contain skin absorption enhancement substance.

Embodiment

Hereinafter, the structure and effect of the present invention will be further illustrated by test example and dosage form example.However, these are tested Example and dosage form example are only to assist in the present invention is understood as purpose is illustrated come what is provided, and scope of the invention and scope are not It is defined in following examples.

The preparation of [reference example 1] ginsenoside Mc

In order to which the ginsenoside Mc that the effect of present composition is tested and used is studied purchased from peace rich (AMBO) Institute.

[test example 1] elastase activity inhibits the measure of effect

Ability is hindered for the elastase activity of ginsenoside Mc, with Epigallo-catechin gallate (EGCG) (EGCG) it is compared and measures.The elastoser and matrix used is commercially public purchased from U.S.'s Sigma-Aldrich Take charge of the elastoser of (Cat.No.E0127).

Elastase activity inhibition is tested with following test methods.

In 96 orifice plates, 10mg/L Tri(Hydroxymethyl) Amino Methane Hydrochlorides (Tris-HCL) buffer solution is being modulated into (pH8.0) in, 20 μ g/mL elastoser-type III solution of the ginsenoside Mc and 50 μ L of 200 μ L are mixed.By 250 μM EGCG is used as positive controls, and the non-process group as negative control group has used distilled water.Afterwards, add N- succinyl-alanine-the Ala-Alas-of the 0.4514mg/mL modulated with the buffer solution of 100 μ L are to nitro acyl Aniline (N-SUCCINYL-ALA-ALA-ALA-p-NITROANILIDE), and reacted 15 minutes at 25 DEG C.Reaction terminates Afterwards, absorbance (synergistic effect 2, the microplate reader (BioTek) (VT, the U.S.) under 415nm wavelength are measured.With identical method come real It applies blank test and is corrected.

The computational methods of elastase activity inhibition the results are shown in table 1 below as shown in following mathematical expressions 1 In.

Mathematical expression 1

Elastase activity obstruction rate (%)=1- (C-D)/(A-B) × 100

A：In no addition substances, and when adding enzyme, the absorbance under 415nm wavelength

B：In no addition substances, enzyme, the absorbance under 415nm wavelength

C：When adding substances, enzyme, the absorbance under 415nm wavelength

D：Addition substances, without add enzyme when, the absorbance under 415nm wavelength

Table 1

Compound	Inhibition level (%)
		Non-process group	0
EGCG	65
		Ginsenoside Mc	75

As shown in above-mentioned table 1, inhibition levels of the ginsenoside Mc to elastase activity is shown, than being known as bullet Property proteinase activity inhibitor EGCG it is significantly excellent, thus it is confirmed that the present invention ginsenoside Mc elastin laminin enzyme activity Property inhibition is excellent.

[test example 2] clostridiopetidase A (MMP-1) hinders ability

Obstruction ability is generated for the clostridiopetidase A of the ginsenoside Mc of the present invention, is measured compared with retinoic acid.

Equipped with DMEM (Dulbecco ' s Modified Eagle ' s Media) trainings containing 2.5% fetal bovine serum In the 96 hole plate incubators (96-well microtiter plate) for supporting base, added in the amount of 5,000 cells/wells (well) Human fibroblasts, and in 5%CO₂, culture is carried out in 37 DEG C of incubators until 70~80% degree are grown to.With 10 μ g/ The ginsenoside Mc of ml concentration or during small retinoic acid treatments 24 after, take cell culture fluid.

Using available commercially as clostridiopetidase A sensing equipment (Amersham Phamarcia companies of the U.S., Catalog#： RPN2610), the clostridiopetidase A generation degree for the cell culture fluid taken is measured.First, once clostridiopetidase A antibody is smeared uniform 96- hole plates (96-well plate) in add in the cell culture fluid taken, and it is anti-to implement in thermostat Ag-Ab Answer 3 it is small when.3 it is small when after, the secondary collagen antibody for being combined with colour developing group is added in 96- hole plates, and secondary response again 15 minutes.After 15 minutes, add in as develop the color evocating substance 3,3 ', 5,5 '-tetramethyl benzidine (3,3 ', 5,5 '- Tetramethylbenzidine, Sigma), and colour developing 15 minutes is induced at room temperature, it adds in 1M sulfuric acid again and stops showing Colour response, then the color of reaction solution is in yellow, and according to the carry out degree of reaction, the degree of the yellow shown is different.

Using absorptiometer, measure is in the absorbance of the 96- hole plates of yellow under 405nm, and according to following mathematical expressions 2 The synthesis degree of clostridiopetidase A is calculated, and the results are shown in table 2 below.At this moment, by the group of never useful compositions-treated In the reaction absorbance of cell culture fluid taken as a control group.

Mathematical expression 2

Absorbance × 100 of absorbance/control group of collagenase expression degree (%)=substance processing groups of cells

Table 2

Compound	Expression degree (%)
		Non-process group	100
Retinoic acid	75
		Ginsenoside Mc	73

As shown in above-mentioned table 2, it is known that the collagenase expression degree of ginsenoside Mc is with being known as collagenase expression The retinoic acid of inhibitor is compared, and collagenase expression hinders effect level similar.

By the above results, it can confirm that the ginsenoside Mc of the present invention has and hinder matrix metalloproteinase (MMP-1) Effect.

[dosage form example 1 and compare dosage form example 1]

According to the composition of Table 3 below, nourishing cream (unit is prepared by conventional method：Weight %).

Table 3

[test example 3] skin elasticity improves effect confirmation

In order to confirm the effect to the raising of the skin elasticity of people, using the dosage form example 1 and the dosage form for comparing dosage form example 1, And following evaluation is made.

For the healthy women of 20 30 to 40 years old age brackets, it is divided into two groups with every group 10, by dosage form example 1 and compares agent After the nourishing cream of 1 two groups of type example is applied to face 12 weeks with frequency 1 time a day, skin elasticity tester is utilized (Cutometer SEM575, C+K Electronics Co., Ltd.s (C+K Electronic Co.), Germany) measures skin elasticity.It is tied Fruit is represented in table 4 below.The end value of table 4 is remembered with the Δ R8 values of Cutometer (Cutometer SEM 575) It carries, wherein R8 values represent the property of skin viscoplasticity (viscoelasticity).

Table 4

Experimental products	Skin elasticity effect
		Dosage form example 1	0.44
Compare dosage form example 1	0.10

As shown in above-mentioned table 4, the dosage form example 1 of the ginsenoside Mc containing the present invention, the group of dosage form example 1 compared with smearing It compares, skin elasticity improves more.

Therefore, it can confirm that the composition of the ginsenoside Mc containing the present invention is highly effective to skin elasticity raising.

[test example 4] improves the confirmation of wrinkle of skin effect

In order to confirm that the composition of the present invention to the effect improving wrinkles of people, make use of the dosage form example 1 and compare dosage form Example 1.

For the effect improving wrinkles for confirming the dosage form example 1 He comparing dosage form example 1, following evaluation has been made.For 20 The healthy women of 40 years old age bracket of name, is divided into two groups with every group 10, by dosage form example 1 and compares the nutrition of 1 two groups of dosage form example After frost is applied to face 12 weeks with frequency 1 time a day, replica (replica) is taken out using silicon, and uses skinanalysis apparatus (visiometer, SV600, Courage+Khazaka electronic GmbH, Germany) measures the state of wrinkle, and carries out Graphical analysis.Its result is represented in table 5 below.The result of table 5 below is represented in each parameter (parameter) after smearing 12 weeks Subtract the average value after the parameter value before smearing.

Table 5

As shown in above-mentioned table 5, it is known that the preparation composition for external use of dosage form example 1 is to improving the effect of wrinkle of skin very It is excellent.

[test example 5] tyrosinase hinders effect

Tyrosinase is the extraction from mushroom class (Mushroom), has used the tyrosinase of Sigma.First, It will be dissolved in as the tyrosine of matrix in distilled water and the solution of 0.3mg/ml be made, and by the solution with every pipe 1.0ml Amount be added in test tube after, wherein add 1.0ml potassium-phosphate buffer solution (0.1mol concentration, pH6.8) and The distilled water of 0.7ml.

Sample liquid is prepared to mix ginsenoside Mc with 10ppm in the ethanol solution of the present invention, by the above-mentioned of 0.2ml Sample liquid is added in reaction solution, is then reacted 10 minutes in 37 DEG C of thermostats.At this moment, 0.2ml solvents will only be added in and carrys out generation For adding in the group of each sample liquid as a control group, and used using ascorbic acid as positive controls.In the reaction solution The tyrosinase solution of 2500 units/ml of 0.1ml is separately added into, and is reacted 10 minutes in 37 DEG C of thermostats again.It will dress The test tube for having the reaction solution, which is put into ice water, makes its rapid cooling, so as to stop reacting, and with photoelectricity spectrum analysis instrument, measures Absorbance (synergy 2, microplate reader (VT, the U.S.), and the results are shown in table 6 below under 475nm wavelength.Under It states mathematical expression 3 and hinders effect to calculate each tyrosinase.

Mathematical expression 3

Tyrosinase obstruction rate (%)=100- (the reaction absorbance of reaction absorbance/control group of substances × 100)

Table 6

Substances	Tyrosinase obstruction rate (%)
		Control group (does not add)	0
Ascorbic acid	52
		Ginsenoside Mc	63

It is recognised that ginsenoside Mc according to the present invention is to the inhibiting rate ratio of tyrosinase from the result of above-mentioned table 6 Ascorbic acid as well known tyrosinase inhibitor is much higher, therefore whitening effect is very excellent.

[test example 6] generates inhibition using the melanin of B16/F10 melanoma cells

To the sample of ginsenoside Mc and kojic acid be contained as substances using the amount of 0.001 weight % respectively, and with one Determine concentration to be added in the culture solution of B16/F10 melanoma cells (bank of Korea Cell system), and cultivate 3 days after, removal training Then nutrient solution is cleaned with phosphate buffer (PBS), and with after 1N NaOH dissolving cells, absorbance (association is measured under 405nm With effect 2, microplate reader (VT, the U.S.).To be not added with the cells of substances as a control group, and with the melanin in control group Content is compared, and measures the degree that each substances hinder melanin generation.Melanin generation suppression is calculated according to mathematical expression 4 Rate processed, and the results are shown in table 7.

Mathematical expression 4

Melanin generating suppression (%)=100- (absorbance × 100 of absorbance/control group of substances)

Table 7

Substances	Melanin generating suppression (%)
		Control group (does not add)	0
Kojic acid	53
		Ginsenoside Mc	66

It is recognised that the inhibiting rate that ginsenoside Mc according to the present invention generates melanin from the result of above-mentioned table 7 It is more much higher than the kojic acid as well known melanin generation Inhibitors, therefore whitening effect is very excellent.

[test example 7] skin moisture-keeping power increases effect measuring

The effect generated is increased to skin moisture-keeping power in order to measure ginsenoside Mc, the dosage form example 1 is make use of and compares Dosage form example 1, and made following evaluation.

For the adult men and women for being categorized as dry skin of 20 40 to 50 years old age brackets, it is divided into two groups with every group 10, Dosage form example 1 and the nourishing cream for comparing 1 two groups of dosage form example are applied to face 4 weeks with frequency 2 times a day.Smearing start before, After smearing 1 week, after 2 weeks, after 4 weeks when and stopping smearing by after 2 weeks (altogether by 6 weeks), in constant temperature, constant humidity condition Under (24 DEG C, relative humidity 40%), moisture of skin tester (Corneometer CM825, C+K Electronics Co., Ltd. (C+ is used K Electronic Co.), Germany) measure moisture of skin amount.Its result is represented in table 8 below.Table 8 the result is that with experiment Start the value of the moisture of skin tester measured before as benchmark, by the increase part of the measured value after processing a period of time Result expressed as a percentage.

Table 8

From the result of the table 8 it has been confirmed that when smearing compares dosage form example 1, until 4 weeks by smearing, show About 30% moisture increment rate is shown, but stops moisture of skin amount after smearing and reduces, on the contrary, smearing the agent containing ginsenoside Mc During type example 1, also major part shows more than 30% moisture of skin increment rate after stopping smearing.It is possible thereby to know containing ginseng The skin moisture-keeping power excellent effect of the composition of the present invention of saponin(e Mc.

[test example 8] Keratinocyte differentiation facilitation effect measures

It is as follows, in order to understand ginsenoside Mc Human Keratinocytes differentiation facilitation effect, using absorbance come Measure hornification coating (Cornified Envelope, the CE) amount generated during Keratinocyte differentiation.

First, the keratinocyte of the people cultivated after being separated from the epidermis of baby and once is put into culture and burns In bottle, after it is made to be attached to bottom, with after the ginsenoside Mc processing of the concentration of 5ppm in culture solution, culture 5 days until Until cell grows to the 70~80% of bottom area.At this point, low calcium (0.03mM) processing group and high calcium (1.2mM) are handled into component It Zuo Wei not negative control group and positive controls.Then the cell of above-mentioned culture is obtained, after being washed with phosphate buffered saline (PBS), Add in the Soviet Union of two sulphur containing 2% lauryl sodium sulfate (sodium dodecyl sulfate, SDS) and 20mM concentration of 1ml The 10mM concentration of sugar alcohol (Dithiothreitol, DTT) Tri(Hydroxymethyl) Amino Methane Hydrochloride buffer solution (Tris-HCl, PH7.4), and (sonication) is ultrasonically treated, (boiling) is boiled, centrifuges, then precipitation is suspended in 1ml In PBS, and measure absorbance (synergy 2, microplate reader (VT, the U.S.) under 340nm.In addition, it takes out a part of described super Solution after sonication measures protein content, as the benchmark of evaluation cell differentiation.And it the results are shown in In table 9 below.

Table 9

Substances	Differentiation capability (%) in keratinocyte
		Low calcium (0.03mM) solution (negative control group)	100
High calcium (1.2mM) solution (positive controls)	210
		Ginsenoside Mc	285

As shown in Table 9 above, it can confirm that the differentiation facilitation effect of keratinocyte is excellent when being handled with ginsenoside Mc It is different.

[test example 9] skin barrier function recovery effects measure

It is real in order to measure the effect that ginsenoside Mc recovers to generate to the skin barrier function impaired due to skin injury Following experiments are applied.For the upper arm of 10 adult men and women, the method that (Tape Stripping) is removed with adhesive tape damages skin Barrier smears the dosage form example 2 prepared according to the composition of table 10 below and compares 2 two groups of dosage form example, uses Vapometer respectively (dolphin (Delfin), Finland) measures the recovery extent of 1 percutaneous moisture loss amount (TWEL), measures seven days, and carry out daily Compare.Here comparison dosage form example 2 is the carrier (vehicle) of negative control group.Tested that the results are shown in table 1 below 1 In.Table 11 the result is that the difference of the before processing before barrier injury and after barrier injury is compared as 100% benchmark As a result.

Table 10

Food ingredient	Dosage form example 2	Compare dosage form example 2
			Purified Water	69	70
Propylene glycol	30	30
			Ginsenoside Mc	1	-

Table 11

It was found from from above-mentioned table 11, it can confirm that when the comparison dosage form example 2 for not containing ginsenoside Mc is used to handle, with The passage of time, percutaneous moisture loss amount gradually increases, on the contrary, when the dosage form example 2 containing ginsenoside Mc is used to handle, Percutaneous moisture loss amount recovers normal with speed quickly, and barrier injury is restored.

[test example 10] color improvement

It is how general using laser in order to evaluate cosmetic composition according to the present invention to promoting the effect that skin blood cycles Strangle blood flow imaging instrument (Laser Doppler Perfusion Imager, LDPI；Periscan PIM II, lark prestige (Perimed) (Stockholm, Sweden)), measure blood circulation degree in skin.Known LDPI is the blood measured in skin The instrument for cycling, and it is now widely used instrument, is a kind of blood that can not only measure in capillary of skin Speed and amount, and the very delicate of the flowing in parteriole and veinlet can be measured.

In thermostatic constant wet chamber, after being washed one's face with perfumed soap, adapt to 30 minutes, and initial value is determined using LDPI.First, It is determined with the initial stage blood flow below the forehead of 30 ice-cold to usually trick LDPI women.Then, experiment pair is made Blood flow is measured afterwards as using 1 week dosage form example 1 and comparing dosage form example 1, then by the blood flow of measure and the initial stage The result (skin blood flow variation) that measured value is compared is represented in table 1 below 2.

Table 12

It has been confirmed that cosmetic composition according to the present invention is not with containing ginsenoside Mc from the result of above-mentioned table 12 Comparison dosage form example 1 compare so that skin blood flow dramatically increases, and is promoted by this blood circulation so that complexion obtains Improve.This finally shows that the cosmetic composition according to the present invention containing ginsenoside Mc can be to effectively transferring skin-nourishing Simultaneously delay skin aging contributes for ingredient and inhibition.

[test example 11] colour of skin improvement

In order to understand the dosage form example 1 and compare the colour of skin improvement of dosage form example 1,30 test objects is made to use respectively After (1 times/day of evening smears 1 week altogether), Facial Stage DM-3 (jasmine spy (Moritex), Japan) instrument evaluation is utilized The colour of skin improves degree.For colour of skin improvement rate, become using the lightness and color measured value of skin and the lightness and color of skin Change value is judged, and the results are shown in table 1 below 3.Lightness and the higher expression colour of skin of color change value are changed It is kind.

Table 13

It has been confirmed that not containing the comparison dosage form example 1 of ginsenoside Mc according to the present invention from the result of table 13, do not show Show that the significant colour of skin improves effect, on the contrary, using containing dosage form examples 1 of the ginsenoside Mc as active ingredient, after use Compared with the colour of skin is before use, it is greatly improved.

[test example 12] pore refining effect

1. the effect of the pore refining of the biosynthesis by promoting collagen

By ginsenoside Mc according to the present invention to the facilitation effect and Peritoneal fibrosis of the biosynthesis of collagen β (TGF-β) is compared and is determined.First, by fibroblast with every hole 10⁵The amount of a cell is inoculated in 24 holes (well) in, and culture is carried out until growing to 90% or so.It is small with serum-free cell culture medium (DMEM) culture 24 When after, respectively using 10ng/ml be dissolved in serum free medium the present invention ginsenoside Mc and TGF-β at Reason, and in CO₂When culture 24 is small in incubator.Their supernatant is taken out, and utilizes procollagen type (I) Enzyme-linked Immunosorbent Assay Measure (ELISA) kit (procollagen type (I) (procollagen type (I)；#MK101, TAKARA (Shiga, Japan) Come whether observing the increase and decrease of procollagen (procollagen), and the results are shown in table 1 below 4.Non-process group is set to 100 and compared for the synthesis capability of collagen.

Table 14

Substances	Collagen synthesis ability (%)
		Non-process group	100
TGF-β	183.5
		Ginsenoside Mc	195.7

It has been confirmed that ginsenoside Mc according to the present invention and positive controls TGF-β phase from the result of above-mentioned table 14 Than showing higher levels of excellent collagen synthesis ability.Therefore, it can confirm that ginsenoside Mc according to the present invention The pore to broaden is shunk by increasing the collagen growing amount on pore periphery.

2. pore refining effect

In order to understand dosage form example 1 and compare the pore refining effect of dosage form example 1, evaluated as follows.Select 20 pores The wide men and women's test object of size is divided into two groups by every group 10, and is smearing dosage form example 1 on the face daily according to group and comparing The nourishing cream of dosage form example 1 is smeared 4 weeks altogether.Judge the effect of pore refining in the following way.Before shooting experiment and after 4 weeks Photo, and allow expert by visually being evaluated.Its result represents the (opinion rating in table 1 below 5：0- is not received completely Contracting；5- shrinks very much).

Table 15

Substances	Opinion rating
		Dosage form example 1	4
Compare dosage form example 1	0

It was found from from the result of above-mentioned table 15, compare effect of the dosage form example 1 without pore refining, however, dosage form example 1 is shown The pore refining effect that can with the naked eye confirm, so as to understand that the ginsenoside Mc of the present invention is excellent to the effect of pore refining size It is different.

[test example 13] sebum secretion inhibition

1. the effect of the inhibition skin hypersecretion by inhibiting 5α-reductase activity

In order to confirm 5α-reductase activity suppression effect, determined in HEK293-5 α R2 cells [¹⁴C] testosterone changes into [¹⁴C] dihydrotestosterone (DHT：Dihydrotestosterone ratio).P3x FLAG-CMV-5 α are transfected on HEK293 cells After R2, and by per hole 2.5 × 10⁵The amount of a cell is inoculated in 24 orifice plates, and cultivated (Park et al., 2003, JDS.Vol.31, pp.191-98).Change within second day the new culture medium added with zymolyte and inhibitor into.By 0.05 μ Ci [¹⁴C] testosterone (kit (Amersham Pharmacia biotech), Britain) be used as culture substrate.

In order to confirm 5α-reductase activity suppression degree, ginsenoside Mc is added in, and at 37 DEG C, 5%CO₂In incubator Cultivate 2 it is small when.At this point, the group for not adding in ginsenoside Mc is used as negative control group, Finasteride will be added in (finasteride) group is used as positive controls.Recycle culture medium afterwards, and with 800 μ l ethyl acetate extraction steroids it Afterwards, the organic solvent layer on top is separated, and after drying, is dissolved to remaining residue, then with 50 μ l ethyl acetate, and in silicon On plastic film silica gel 60F254 (Silica plastic sheet kieselgel60F254), ethylacetate-hexane is used (1:1) it is unfolded as solvent.

By plastics sample after being dried in air, shower system has been used in order to measure the amount of isotopeDry sheet plastic and x-ray film are added to bath box togetherIn, 1 The isotopic mass for staying in testosterone and dihydrotestosterone on film is measured after week, then according to following mathematical expressions 5 and 6, is calculated respectively Conversion ratio and obstruction rate, and the results are shown in table 1 below 6.

Mathematical expression 5

Radiant/total radiant × 100 in conversion ratio (%)=DHT regions

Mathematical expression 6

Conversion ratio × 100 of inhibiting rate (%)=[conversion ratios of conversion ratio-substances of control group]/control group

Table 16

Substances	Conversion ratio (%)	Obstruction rate (%)
			Negative control group	48.0	-
Positive controls	27.6	42.5
			Ginsenoside Mc	14.8	67

It has been confirmed that ginsenoside Mc can effectively inhibit the activity of 5α-reductase from above-mentioned 16 result of table, so as to Testosterone is blocked to be converted into dihydrotestosterone, and is shown compared with the Finasteride of known inhibition 5α-reductase activity more Add excellent inhibition.The 5α-reductase makes testosterone be converted into dihydrotestosterone, so as to intracytoplasmic receptor protein knot It closes and enters in core, so as to activate sebocyte cell and promote to break up, so that the sebum excessive secretion in sebaceous glands.Cause This, it is thus identified that ginsenoside Mc inhibits the excessive secretion of sebum by effectively inhibiting the activity of 5α-reductase.

2. sebum secretion inhibition

In order to understand the dosage form example 1 and compare the sebum secretion inhibition of dosage form example 1, following evaluation has been carried out.Choosing Go out 30 men and women's test objects thought more than sebum secretion, smear dosage form example 1 daily in the appointed part of skin on the face and compare The nourishing cream of dosage form example 1 is smeared 4 weeks altogether.Judgement for sebum minimizing effect, by using sebum tester (Sebumeter SM810, C+K Electronics Co., Ltd.s (C+K Electronic Co.), Germany) is measured respectively by 2 weeks and 4 Average sebum slip (%) after week, and the results are shown in table 1 below 7.

Table 17

It was found from from the result of above-mentioned table 17, it is of the invention contain ginsenoside Mc as active ingredient dosage form example 1 with Comparison dosage form example 1 without it is compared, and can effectively inhibit the sebum excessively secreted.

[dosage form example 3 and compare dosage form example 3~4]

It prepares dosage form example 3 according to the ingredient and content (weight %) shown in table 1 below 8 and compares dosage form example 3~4, It is described as follows.Dosage form example 3 is the substance of mixing ginsenoside Mc, compares dosage form example 3 to improve completely without bag containing acne The substance of the active ingredient of skin, standard substance of the comparative example 4 as the benchmark for antimicrbial power, which contains is used as Cuo more The erythromycin (erythromycin) that sore therapeutic agent uses.

Dosage form example 3 and compare dosage form example 3~4 preparation method it is as follows.The ingredient of the A items of table 1 below 8 is completely dissolved, And in other dissolving tanks, the ingredient of B is completely dissolved, B are added in A afterwards, makes its mixing solubilized.And Wherein, the ingredient of C is added with the mixed proportion according to described in table 18, and after mixing, is filtered, so as to prepare This composition.

Table 18

[test example 14] tests the antimicrbial power of acne bacterium

Using each cosmetic composition according to the dosage form example 3 and the composition preparation for comparing dosage form example 3~4, to conduct Propionibacterium acnes (the ATCC6919 of acne pathogenic strain：Culture medium-brain heart infusion broth culture medium (BHI broth))) into Row antimicrbial power is tested.

It is as follows to the antimicrbial power test method of acne bacterium.

(1) preparation of test bacteria liquid

Using using propionibacterium acnes be inoculated in brain heart infusion broth culture medium carry out Anaerobic culturel culture solution as Test bacteria liquid.

(2) preparation of dilute solution

The addition 0.15ml in the brain heart infusion broth culture medium (pH6.8) or LB broth bouillons (pH4.5) of 15ml The test bacteria liquid, and use using the mixed liquor mixed as dilute solution.

(3) preparation of sample

By dosage form example 3 and the cosmetic composition stoste prepared in dosage form example 3~4 will be compared, made directly as sample With.

(4) antimicrbial power is tested

1) sample is added in No. 1 row of cell culture tube (96well plate) in 96 holes so that it is matched with initial concentration, And adding in dilute solution makes total amount be 200 μ l.

2) by the mixed liquor of No. 1 row after mixing, take out the mixed liquor of 100 μ l and be added in No. 2 rows, and mix equal After even, the mixed liquor of 100 μ l is taken out again and is added in No. 3 rows.Secondary dilution (double is carried out in this way dilution)。

3) after when quiescent culture 24 is small at 32 DEG C and when 48 is small, judge whether bacterium rises in value with suspension degree, and will The Cmin for not having the multiplication of bacterium is set to minimum inhibitory concentration (Minimum Inhibitory Concentration, MIC) Value.It is difficult to judge whether bacterium rises in value if mixed liquor is opaque, confirm by using micro- sem observation.

The antimicrbial power result of the test of acne bacterium is represented in table 1 below 9.For minimum inhibitory concentration, dosage form is converted into In the concentration of active ingredient that contains and mark.

Table 19

Project	pH	Propionibacterium acnes
			Dosage form example 3	5.7	>52ppm
Compare dosage form example 3	5.7	Maximum concentration (does not have antimicrbial power)
			Compare dosage form example 4	5.7	>100ppm

In the result of table 19, in minimum inhibitory concentration, ppm concentration is smaller, it is believed that the substance is to acne bacterium The effective substance of antimicrbial power, during using dosage form example 3, the comparison dosage form with the erythromycin used as well known acne therapeutic agent Example 4 is compared, and the ppm concentration shown is significantly low, whereby it was confirmed that the composition containing ginsenoside Mc has test organisms Superior antimicrbial power.

[test example 15] lipid synthesis (Lipogenesis) inhibits experiment

Using as the 3T3-L1 cells of the fibroblast of mouse (fibroblast cell line), with 1 × 10⁵Carefully Born of the same parents/hole, which is attached at, fills the DMEM (Dulbecco ' s containing 10% fetal bovine serum (fetal Bovine Serum, FBS) Modified eagle ' s medium, GIBCO BRL, live technology company) culture medium 6 well culture plate (culture Plate in).After 2 days, new DMEM (containing 10% FBS) culture medium is re-replaced, and is cultivated 2 days.Then, with containing There are 1 μ g/ml insulin (insulin), 0.5mM isobutyl methylxanthines (IBMX) and 0.25 μm of dexamethasone (dexamethasone) DMEM (containing 10% FBS) carries out induction to the cell of the culture, and with 50 μM Then ginsenoside Mc and caffeine processing, after 2 days, re-replace into the DMEM comprising insulin, and cultivate 5 days.5 After it, normal incubation medium (DMEM contains 10% FBS) is re-replaced into, and the cell is observed and is cultivated to institute It states cell and changes lipoblast from form.

In order to evaluate the effect of ginsenoside Mc is to inhibiting Fat Accumulation in adipocyte, complete what is broken up using described 3T3-L1 adipocytes are implemented the Sudan three and are dyed (S4136, Sigma-Aldrich).At normal temperatures, in phosphate buffer In, after 4% paraformaldehyde (pH7.2) fixed fat cell, washed with phosphate buffer, then, with the Sudan three into Photo is shot after row dyeing, and passes through naked eyes and is compared.It will be not added with substances or compare substance and culture medium is used only Group use as a control group, for other comparative groups, handled with 50 μM of caffeines.Fat Accumulation inhibition level is to pass through The degree of dyeing is divided into +++, ++ ,+,-, so as to assign grade, at this point, closer +++, represent that dye levels are bigger.It is tied Fruit is represented in table 2 below 0.

Table 20

As shown in above-mentioned table 20, the ginsenoside Mc used in the present invention can confirm that, not only accumulation in adipocyte Fat mass it is few, and compared with as the caffeine of well known lipid synthesis inhibiting substances, also with excellent lipid synthesis Inhibition.Therefore, sebum is reduced by inhibiting lipid synthesis, so as to inhibit the generation of acne.

[test example 16] acne improves and sebum secretion reduces and stimulate the experiment whetheing there is

Using 30 people with acne as subjects, it is divided into three groups, and the examination to corresponding to each group with every group 10 Object is tested using with the dosage form example 3 and comparing dosage form example 3~4 cosmetic composition for preparing 1 month.Improve for acne Standard, is set as 1 point to 5 points, and mark 1 divides for " not having ", and 3 points are " common ", and 5 points are " very good ".For experimental result, with The average mark of 10 is marked in table 2 below 1.

Period is eliminated for acne, to recognize the number of days of elimination as benchmark, for acne recurrence, to have after 1 month Without recurrence as benchmark.Reduction for sebum secretion is set as 1 point to 5 points, and is marked as 1 point as " not having ", and 3 points are " general It is logical ", 5 points are " very good ".For experimental result, marked with the average mark of 10 in table 2 below 1.With (showing stimulates instead The number answered)/(overall test number) judge the presence or absence of skin irritatin.

Table 21

As shown in above-mentioned table 21, it is known that the dosage form example 3 compared with of dosage form example 3 is compared, acne does not recur, and whole Acne, which is improved, on body has excellent effect.In addition, during using comparison dosage form example 4 containing antimicrbial power standard substance, although It is strong to the stimulation of skin when showing acne improvement, but using the substance, therefore be not suitable for long-time service, however, Composition according to the present invention does not stimulate but, thus it is shown that also being adapted for using for a long time.

[test example 17] inflammation improvement-interleukin 8 (IL-8) generation inhibition

Before one day tested, by Keratoderma epithelial cell (Normal human skin keratinocyte, NHEK buys place：Lonza), with 5 × 10⁴Cells/well is inoculated in 96 orifice plates, then at 37 DEG C, 5%CO₂Incubator (incubator) when culture 24 is small in.24 it is small when after, clean cell 2 times with phosphate buffer, and be replaced with serum-free angling Cell basal culture medium (serum free keratinocyte basement media (KBM)).In each hole, use respectively The ginsenoside Mc processing of 5ppm, 25ppm and 50ppm concentration, and react 30 minutes after, use staphylococcus aureus respectively Peptide glycan (PGSA) (10 μ g/ml), aureus peptide glycan (50 μ g/ml) and aureus peptide glycan (50 μ G/ml)+lipopolysaccharides (1 μ g/ml) processing.Wherein, aureus peptide glycan (PGSA：peptidoglycan from S.aureus it is the cell of Gram-positive (+) bacterium) as the peptide glycan (peptidoglycan) extracted from staphylococcus The main constituents of wall, and the cell membrane component of known bacterium can induce inflammation.According to the report, especially staphylococcus, allergy Property dermatitis patients 90% or so because the bacterium generate superinfection.Known lipopolysaccharides (LPS：Lypopolysaccaride it is) leather The main constituents of the cell membrane of Lan Shi feminine gender (-) bacterium, and be the main reason for inflammation induces.

At 37 DEG C, 5%CO₂After when culture 24 is small in incubator, takes out culture solution and carry out to interleukin 8 The enzyme-linked immunosorbent assay (ELISA) of (Interleukin-8, IL-8), and the results are shown in table 2 below 2.For ELISA make use of the experimental method of preparation company (BD science).

Table 22

Classification	The secretion (pg/ml) of interleukin 8
		Without processing control group (control)	935.12
PGSA(10μg/ml)	4812.60
		PGSA(50μg/ml)	5895.08
PGSA(50μg/ml)+LPS(1μg/ml)	6814.91
		Ginsenoside Mc (5ppm)	1573.32
Ginsenoside Mc (25ppm)	1203.54
		Ginsenoside Mc (50ppm)	1001.23

It has been confirmed that ginsenoside Mc can be substantially reduced and inhibited due to PGSA and lipopolysaccharides LPS from above-mentioned table 22 The secretion of increased interleukin 8.As a result it will be appreciated that the present invention Dermatologic preparation composition can substantially reduce because The secretion of PGSA and LPS and increased interleukin 8, so as to provide excellent anti-inflammatory effect.

[test example 18] itch alleviates evaluation

Before one day tested, by keratinocyte (cell line title：HaCaT buys place：US mode culture Object preservation institute (ATCC)), with 4 × 10⁴Cells/well is inoculated in 96 orifice plates, then at 37 DEG C, 5%CO₂24 are cultivated in incubator Hour.24 it is small when after, it is clear with balanced salt solution (Hanks ' Balanced Salt solution, HBSS) buffer solution of hanks After washing (washing) 96 orifice plate 2 times, by reaction buffer (2 μM of Fluo-4-AM, 20% pluronic acid (pluronic Acid), 2.5mM probenecid (probenecid)) it is put into cell.At 37 DEG C, 5%CO₂It is reacted 30 minutes in incubator, and After reacting 30 minutes at normal temperatures, with HBSS buffer solution for cleaning 2 times, and (weighed with 0.05%, 0.1%, 0.5% and 1.0% concentration Amount %) ginsenoside Mc handle cell.

Reaction after ten minutes, is carried out with the trypsase (trypsin) of 2U/ml or 5 μM of PAR-2 active peptides (SLIGKV) Processing, and measure intracellular Ca²⁺Concentration changes 80 seconds.For intracellular Ca²⁺The measure of concentration variation, make use of microplate reader 3 (FlexStation3：Molecular device (Molecular Device), the U.S.).With the trypsase of ginsenoside Mc and 2U/ml (trypsin) or after 5 μM of PAR-2 active peptides (SLIGKV) processing, the bending (flex) of 80 seconds is measured, and be obtained After the minimum value of value and the difference of maximum, by the value with using 2U/ml trypsase or 5 μM of PAR-2 active peptides (SLIGKV) difference of minimum value and maximum when handling is compared, and intracellular inhibiting rate (%) table is poured in calcium ion Show in table 2 below 3.

Table 23

It is recognised that the calcium ion caused by trypsase or PAR-2 active peptides (SLIGKV) is to thin from above-mentioned table 23 Intracellular pours in, and is reduced with the processing of ginsenoside Mc, and can confirm that with the concentration for improving ginsenoside Mc, calcium Ion is substantially reduced to intracellular pouring in.

Therefore, the Dermatologic preparation composition of the ginsenoside Mc containing the present invention, by effectively inhibiting to induce itch PAR-2 activity, so as to provide excellent antipruritic effect.

[dosage form example 4 and compare dosage form example 5]

Shampoo is prepared with the composition of table 2 below 4.Specifically, surfactant and glycol distearate are added It is added in Purified Water, and after being heated to 80 DEG C and making its uniform dissolution, is cooled to 40 DEG C gradually under stiring, also, described After active ingredient according to the present invention and preservative, viscosity modifier, fragrance and hair conditioner are added in mixture and is mixed, It is cooled to room temperature under stiring, so as to prepare shampoo.

Table 24

Ingredient	Dosage form example 4	Compare dosage form example 5
			Ammonium lauryl sulfate	10	10
Polyoxyethylene lauryl base ammonium sulfate	5	5
			Cocoamidopropyl betaine	2	2
Glycol distearate	1.5	1.5
			Cocomonoethanolamide	0.8	0.8
Ginsenoside Mc	5.0	-
			Polyquaternium-10	0.2	0.2
Blueness 1	0.0002	0.0002
			Yellow 4	0.0001	0.0001
Methyl p-hydroxybenzoate	0.1	0.1
			Fragrance	0.8	0.8
Citric acid	0.1	0.1
			Dimethicone	1.0	1.0
Water	To 100	To 100

[test example 19] dandruff minimizing effect is tested

The more male of 19 to 35 years old of 24 dandruffs is selected, is divided into two groups with every group 12, and divides with the following methods Not using dosage form example 4 and after comparing the shampoo 1 month of dosage form example 5, dandruff slip is measured.

Before on-test, hair usually is cleaned with conventional shampoo, the dandruff for then accumulating two days after acquisition hair washing, And by the weight of the dandruff of acquisition with respectively with dosage form example 4 and compared with dosage form example 5 shampoo washed every two days a hair and The weight for the dandruff for accumulating two days after the test is compared and evaluates.At this point, the scalp crumb vacuum suck by accumulation Device is directly gathered from scalp, and dandruff slip is obtained according to following mathematical expressions 7, and the results are shown in following tables In 25.

Mathematical expression 7

Dandruff slip (%)=(dandruff weight of the dandruff weight (mg) after-one month before on-test (mg)) the dandruff weight (mg) × 100 before/on-test

Table 25

It is recognised that the dosage form example 4 containing ginsenoside Mc shows that excellent dandruff prevents from imitating from above-mentioned table 26 Fruit.

[test example 20] pruritus of scalp prevents the experiment of effect

Selected 24 than the more serious men and women of 25 years old to 45 years old for feeling itching of the scalp, are divided into two groups with every group 12, And, pass through following evaluation criteria pair with each dosage form example 4 of frequency usage once three days and the shampoo for comparing dosage form example 5 after two weeks Pruritus of scalp prevents effect from being evaluated, and the results are shown in table 2 below 6.

[evaluation criteria]

It is -5 points very excellent

Excellent -4 points

Generally -3 points

Bad -2 points

It is -1 point very bad

Table 26

Classification	Dosage form example 4	Compare dosage form example 5
			The removal effect of pruritus of scalp	4.2	2.3

It is recognised that the dosage form example 4 containing ginsenoside Mc prevents from showing to pruritus of scalp from above-mentioned table 26 Superior effect.

[test example 21] potassium ion channel activity increases effect assessment

Minoxidil as alopeciaing therapeutic agent is known as potential mitochondria K ~+Channel Opener (K_ATP Channel opener), it is the representative drugs used in the treatment of androgenetic alopecia.In order to evaluate this minoxidil Mechanism, used following test method(s)s.The test method(s) is the prevention K in the fibroblast using composition scalp corium_ATPIt is logical The orinase (SIGMA AIDRICH, T0891) in road is handled, and so as to inhibit the multiplication of cell, and is again turned on potassium Ion channel and recover cell Proliferation.

In order to evaluate the K as this composition_ATPThe function of channel opener, present invention uses as fibroblast Mouse embryonic fibroblasts system (Mouse embryonic fibroblast the cell line, NIH3T3 of system：).This cell It is for 3T3 schemes (protocol), to separated into fiber from NIH Swiss mouse embryos (Swiss mouse embryo) Cell line carries out cell line obtained from nature immortalization.The cell line, containing 10%FBS DMEM (Gibco BRL, Gaithersburg, MD, the U.S.) in, maintaining 5%CO₂, when culture 24 is small in 37 DEG C of incubator.NIH3T3 is inoculated in 96 holes In plate, and when culture 24 is small in 37 DEG C of incubator after, handled with the orinase of 2.5mM, and at 10 minutes Afterwards, serve as 10 μM of minoxidil of positive controls and the ginsenoside Mc of 2.5ppm, 5ppm and 10ppm concentration into Row processing, and after drug-treated when 48 is small after, WST-1 kits (Roche (Roche)) is used to measure cell Proliferation energy Power.As a result represent in table 2 below 7.

Table 27

Classification	Ability of cell proliferation (%)
		Without processing control group (control)	100
Minoxidil	132
		Ginsenoside Mc (2.5ppm)	115
Ginsenoside Mc (5ppm)	123
		Ginsenoside Mc (10ppm)	130

From above-mentioned table 27 it is recognised that when being handled with ginsenoside Mc, fibroblastic multiplication is restored, and Ability of cell proliferation increases dependent on the concentration of the ginsenoside Mc of processing, and can confirm that with 10ppm ginsenosides Mc During processing, cell Proliferation is restored to level when being handled with minoxidil.

The melanin generation facilitation effect experiment of [test example 22] ginsenoside Mc

The penicillin of the fetal bovine serum of addition 5%, 100IU in serum-free cell freezing media (RPMI culture mediums) In the culture medium of G and 0.2 μM of terephthalic acid (TPA) (TPA), by melanocyte, with 50,000 cells/wells, 24 orifice plates are inoculated in In.Second day, to the cell of inoculation, by the use of at the ginsenoside Mc as substances of the ultimate density of 10ppm or 50ppm Reason.It, will be at 100 μM of isobutyl methylxanthine (IBMX) using the group handled by the use of 0.1% DMSO as negative control group The group of reason cultivates above-mentioned each group three days as positive controls at a temperature of 37 DEG C.After culture, phosphate buffer is used (PBS) cleaning orifice, and after adding in the 1N NaOH of 100 μ l in every hole, dissolve the melanin in cell.Utilize tablet culture Analyzer (microplate reader) measures absorbance (synergistic effect 2, the enzyme mark of dissolved melanin under 405nm Instrument (VT, the U.S.).Result of the melanin generation facilitation effect of ginsenoside Mc compared with control group is represented following In table 28.

Table 28

Sample	B16 cell amount (%)
		DMSO (0.1%)	100
IBMX(100μM)	120
		Ginsenoside Mc (10ppm)	112
Ginsenoside Mc (50ppm)	121

It is recognised that ginsenoside Mc promotes the B16 cell of melanocyte, so as to increase black from above-mentioned table 28 The generation of element, therefore can show that excellent melanin generation facilitation effect.

Promotion transcription factors (MITF) and tyrosinase expression of [test example 23] the ginsenoside Mc in melanocyte Effect

Using 501mel cell lines, with 500,000 cells/wells, it is inoculated in 6 orifice plates, and in each hole, with 0.1% Dimethyl sulfoxide (DMSO) (DMSO) processing conduct negative control group, by the use of 100 μM of IBMX processing as positive controls and By the use of the life saponin(e Mc of 10ppm handle as experimental group, and cultivated at a temperature of 37 DEG C 24 it is small when, 48 it is small when and 72 it is small when Afterwards, protein is obtained.For the protein so obtained, western blot (Western is carried out using MITF and tyrosinase Blot) method is tested.Protein Extraction and western blot are implemented by the usually used standard method of those skilled in the art. After implementing western blot, the negative control group in its result is set to 100, and is represented compared with the value in table 2 below 9 In.

Table 29

As shown in above-mentioned table 29, it can confirm that ginsenoside Mc improves MITF and tyrosinase protein in melanocyte The expression of matter.

The antimicrbial power evaluation of [test example 24] ginsenoside Mc

In order to evaluate the antimicrbial power of ginsenoside Mc, antibacterial experiment is implemented.Specific test method is as described below.

Staphylococcus aureus (Staphylococcus aureus), the Escherichia coli used in experiment (Escherichia coli) and pseudomonas aeruginosa (Pseudomonas aeruginosa) are in trypticase soya broth Culture in culture medium (Tryptic Soy Broth)；Candida albicans (Candida albicans) and aspergillus niger (Aspergillus niger) is the culture in Sabouraud dextrose broth bouillon (Sabouraud Dextrose Broth). By culture solution in each culture medium with the dilution that 1/100 (albicans strain is using 1/10) is diluted as test organisms Liquid uses.For aspergillus niger, 2 × 10 will be prepared into⁸The spore suspension of cfu/ml is as test bacteria liquid.

The mixed liquor that the test bacteria liquid that 0.15ml is added in each culture medium of 15ml is mixed is as dilute solution To use.

In No. 1 row of 96 orifice plate, the ginsenoside Mc of 10ppm is separately added into the amount of every 16 μ l of hole, and is separately added into The dilute solution of 184 μ l.The dilute solution of 100 μ l is separately added into other holes.The mixed liquor of No. 1 row is uniformly mixed Afterwards, the mixed liquor for taking out 100 μ l is added in No. 2 rows, and after mixing, the mixed liquor for taking out 100 μ l again is added to No. 3 In row.Doubling dilution has been carried out in this way.

Staphylococcus aureus, Escherichia coli and pseudomonas aeruginosa are cultivated in 32 DEG C of thermostat；Candida albicans Bacterium and aspergillus niger are cultivated in 25 DEG C of thermostat.

48 it is small when after, with suspensibility and microscope come whether confirming the increment of bacterium, so as to determine minimum inhibitory concentration (MIC) Value, and the results are shown in Table 3 below 0.

Table 30

As shown in above-mentioned table 30, it can confirm that ginsenoside Mc shows antimicrbial power to a variety of bacterial strains, and pass through this It can predict that ginsenoside Mc can work in composition as natural antiseptic agent or antiseptic a bit.

Claims (14)

1. the cosmetic combinations of improvement acne are being prepared as the Dermatologic preparation composition of active ingredient comprising ginsenoside Mc Application in object.

2. applications of the ginsenoside Mc as unique active ingredient in the cosmetic composition for improving acne is prepared.

3. the cosmetics of offer antibacterial action are being prepared as the Dermatologic preparation composition of active ingredient comprising ginsenoside Mc Application in composition.

4. applications of the ginsenoside Mc as unique active ingredient in the cosmetic composition for providing antibacterial action is prepared.

5. the cosmetics of offer anti-inflammatory effect are being prepared as the Dermatologic preparation composition of active ingredient comprising ginsenoside Mc Application in composition.

6. applications of the ginsenoside Mc as unique active ingredient in the cosmetic composition for providing anti-inflammatory effect is prepared.

7. the makeup of improvement allergic skin is being prepared as the Dermatologic preparation composition of active ingredient comprising ginsenoside Mc Application in product composition.

8. applications of the ginsenoside Mc as unique active ingredient in the cosmetic composition for improving allergic skin is prepared.

9. the cosmetic combinations of pore refining are being prepared as the Dermatologic preparation composition of active ingredient comprising ginsenoside Mc Application in object.

10. applications of the ginsenoside Mc as unique active ingredient in the cosmetic composition for preparing pore refining.

11. the cosmetic combinations of adjusting sebum are being prepared as the Dermatologic preparation composition of active ingredient comprising ginsenoside Mc Application in object.

12. applications of the ginsenoside Mc as unique active ingredient in the cosmetic composition for adjusting sebum is prepared.

13. the change of offer anti-dandruff effect is being prepared as the Dermatologic preparation composition of active ingredient comprising ginsenoside Mc Application in cosmetic compositions.

14. ginsenoside Mc answering in the cosmetic composition that anti-dandruff effect is provided is prepared as unique active ingredient With.