CN105246489A

CN105246489A - Topical composition for skin containing gincenoside RF

Info

Publication number: CN105246489A
Application number: CN201480029495.9A
Authority: CN
Inventors: 金东泫; 柳权烈; 李沃澯; 廉明勋; 曺濬喆
Original assignee: Amorepacific Corp
Current assignee: Amorepacific Corp
Priority date: 2013-04-24
Filing date: 2014-04-24
Publication date: 2016-01-13
Anticipated expiration: 2034-04-24
Also published as: KR20140126891A; CN107898656B; HK1213772A1; CN108498365A; CN108498365B; TW201521735A; CN108042386A; CN105246489B; WO2014175676A1; CN107898656A; TWI620567B; KR102024572B1

Abstract

The present invention relates to a composition containing, as an active ingredient, gincenoside RF, for providing effects such as improvement of acne and skin troubles, skin clearing, pore tightening, in addition to providing effects such as skin tone improvement, hair growth promotion, improvement of graying hair, anti-dandruff, and antisepsis.

Description

Dermatologic preparation composition containing ginsenoside RF

Technical field

The present invention relates to a kind of compositions, described compositions contains ginsenoside Rf (GinsenosideRf), thus effect, convergence skin and the effect of pore refining of improving acne and skin problem can be provided, and can provide improve skin color effect, promote hair growth, improve poliosis, anti-dandruff and antiseptic effect.

Background technology

Skin, as the first defensive barrier of human body, has the stimulation of the external environment condition making intracorporeal organ from the change of such as temperature and humidity, ultraviolet, public hazards material and protects the function of intracorporeal organ.With advancing age, skin will change due to inherent, extrinsic factor.That is, from inherent aspect, owing to regulating metabolic various hormone secretion to reduce, and the function of immunocyte and cytoactive will reduce, and therefore the immune protein of needed by human body and the biosynthesis of organism constitutive protein will reduce.From external aspect, due to the destruction of ozone layer, the ultraviolet content arriving earth's surface from sunray will increase, and along with the in-depth of environmental pollution, free radical and active oxygen will increase, not only make that the thickness of skin reduces thus, wrinkle will increase, elastic force reduces, and make skin complexion dimmed, always there is problem (trouble) in skin, and nevus and freckle and senile plaque are also increased, thus cause the various changes such as complexion is deteriorated, the colour of skin is dimmed.

In order to the skin condition change preventing these from producing due to skin inherence and extrinsic factor, and maintaining healthy skin condition, people make great efforts to be used for improving skin condition by adding in cosmetics to enforcement of going forward side by side such as the biological active substances obtained from existing various animal, plant, microorganism etc. always.

Ginsenoside Rf is included in the one of the Saponin in Radix Ginseng, known physiologically active, the suppression voltage dependent channel (Nah demonstrating the effect of the cerebral dysgenesis that anti-lipid peroxidation effect is induced by ethanol with defence etc., etal., Proc.Natl.Acad.Sci.USA, 92,8739-8743,1995) and anti-pain effect (Shinetal., submited, 1997).But to containing ginsenoside Rf as the effect of the improvement acne of the compositions of effective ingredient and skin problem, convergence skin and pore refining effect, improve skin color effect, promote hair growth, improve poliosis, anti-dandruff and antiseptic effect also do not report.

Summary of the invention

The technical problem solved

To this, the present inventor finds that ginsenoside Rf can provide the effect of effect, convergence skin and the pore refining improving acne and skin problem, and can provide improve skin color effect, promote hair growth, improve poliosis, anti-dandruff and antiseptic effect, thus complete the present invention.

Therefore, the object of the invention is to, a kind of Dermatologic preparation composition is provided, described compositions contains ginsenoside Rf, thus effect, convergence skin and the effect of pore refining of improving acne and skin problem can be provided, and demonstrate improve skin color effect, promote hair growth, improve poliosis, anti-dandruff and antiseptic effect.

Technical scheme

To achieve these goals, the invention provides a kind of containing ginsenoside Rf as the Dermatologic preparation composition for improving acne of effective ingredient.

In addition, the invention provides a kind of containing ginsenoside Rf as the Dermatologic preparation composition for improving color and the colour of skin of effective ingredient.

In addition, the invention provides a kind of containing ginsenoside Rf as the Dermatologic preparation composition for pore refining of effective ingredient.

In addition, the invention provides a kind of containing ginsenoside Rf as effective ingredient for trichogenous Dermatologic preparation composition.

In addition, the invention provides a kind of containing ginsenoside Rf as the Dermatologic preparation composition for preventing poliosis of effective ingredient.

In addition, the invention provides a kind of containing ginsenoside Rf as the Dermatologic preparation composition for anti-dandruff of effective ingredient.

In addition, the invention provides the Dermatologic preparation composition that as natural antiseptic agent use of a kind of ginsenoside Rf of containing as effective ingredient.

Beneficial effect

Compositions of the present invention contains ginsenoside Rf, thus antiinflammatory can be provided, improve the effect of acne and skin problem, the effect of convergence skin and pore refining, and can provide improve skin color effect, promote hair growth, improve poliosis, anti-dandruff and antiseptic effect.

Preferred forms

Compositions according to the present invention contains ginsenoside Rf as effective ingredient.

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

Chemical formula 1

Ginsenoside Rf of the present invention can extract from plant, also can synthesize according to method as known in the art and use, also can use the ginsenoside Rf of mercantile-type sale.In addition, ginsenoside Rf can obtain from Radix Ginseng extract.The kind of the Radix Ginseng at this moment used is not particularly limited, and can use water ginseng, Radix Ginseng Rubra, Radix Ginseng, Tai Ji ginseng and tail ginseng etc.And, described Radix Ginseng extract not only comprises by lixiviate in Radix Ginseng, decocts and carry and the leachate obtained, also comprise the part of leachate or wholely carry out concentrated and concentrate that is that obtain or impregnating, decoct, tablet prepared by described concentrated solution after drying again flowing extracts, and be included in Radix Ginseng the chemical substance playing main efficacy results, but also comprise plant itself, and the extract in all parts of the Radix Ginsengs such as stem, root, leaf, flower, fruit can be used, the extract of a certain specific part can't be defined in.In addition, the method extracting ginsenoside Rf from Radix Ginseng extract can use known method.

Particularly, described ginsenoside Rf by method as known in the art, after using water or organic solvent to prepare Radix Ginseng extract, therefrom can be isolated.The organic solvent used in the present invention can be selected from the mixed solvent of ethanol, methanol, butanols, ether, ethyl acetate, chloroform and these organic solvents and water, preferably uses the ethanol of 80%.At this moment, Extracting temperature is preferably 10 ~ 80 DEG C, and can extract 3 ~ 24 hours.If exceed the scope of described Extracting temperature and extraction time, then extraction efficiency can reduce, or the change of composition can occur.

Compositions of the present invention is preferably containing the described ginsenoside Rf counting 0.001 to 50 % by weight with composition total weight.This is due to when the content of described effective ingredient is less than 0.001 % by weight, there is the problem of effect and the low effort brought by described composition; When more than 50 % by weight, there is the problem in cutaneous safety or dosage form.

Compositions of the present invention can use as the Dermatologic preparation composition for improving acne, and its antibacterial effect is excellent, especially excellent to the antibacterial effect of acne pathogenic bacterium, and provides antiphlogistic effects.

Compositions of the present invention can use as the Dermatologic preparation composition for improving color and the colour of skin, when being used in skin, by expansion blood capillary, and blood circulation promoting come smoothly by nutrition supply to skin, and suppress skin aging, therefore, the effect improving color and the colour of skin is remarkable.

Compositions of the present invention can use as pore refining, the Dermatologic preparation composition that regulates sebum and improve skin problem, when being used in skin, suppress the sebum of excessive secretion, and by promoting that the elimination of active oxygen and the synthesis of collagen protein carry out pore refining, and due to the minimizing of inflammatory Cytokines Expression, therefore, suppress the effect of skin problem remarkable.

Compositions of the present invention can use as trichogenous compositions, it is by promoting that resting stage is converted into the anagen hair cycle hair cycle, thus promote the growth of hair, and not only promote the generation of new hair, but also existing hair health is grown, and provide and prevent and the phenomenon suppressing hair to come off from scalp, hypotrichosis or the effect of state that attenuates.

Compositions of the present invention can as preventing from the compositions of poliosis to use, it carrys out activation of melanocyte by the expression increasing the transcription factor (MITF) in melanocyte, and promote melanic synthesis, thus provide advance preventing bringing out and promoting to bring out the effect of hair color poliosis.

Compositions of the present invention can as preventing from the Dermatologic preparation composition of the dandruff to use, it purifies scalp by effectively discharging the toxin being accumulated in hair and scalp, and scalp inflammation can be prevented to react by suppressing the propagation of dandruff bacterium and growth, and, due to the generation of inhibit activities oxygen and the anti-oxidation function brilliance of effect, calm therefore, it is possible to provide and strengthen scalp, and strengthening the effect of intrinsic phylactic power defensive power.

Compositions of the present invention can use as natural antiseptic agent compositions, because compositions of the present invention is natural component, therefore provides the effect that antiseptic effect is remarkable and harmless.

According to compositions of the present invention can containing acceptable carrier on cosmeceutical or Dermatology or base material dosage form.Its whole dosage forms that can use as applicable local provide, and such as, can be provided as solution, gel, solid, the anhydrous product of pasty state in aqueous phase, disperse oil phase and obtain emulsion, suspension, microemulsion, microcapsule, subparticle ball or ion-type (liposome) and nonionic the form of folliculus dispersant, or frost, astringent, lotion, powder, ointment, spray or hide the form of flaw rod.Further, can with foam (foam) form or comprise further compression propellant aerosol combination form use.These compositionss can be prepared according to the method that this area is conventional.

Especially, Dermatologic preparation composition of the present invention is used for preventing the dandruff, for hair growth or for preventing poliosis time, can as the compositions for scalp and hair dosage form, dosage form is not particularly limited, such as, can be turned to hair oil, trichotrophy astringent, scalp care agent, hair-care agent, shampoo, hair conditioner, hair lotion or hair of scalp dual-purpose nursing agent etc. by dosage form.

And, fatty material can be comprised according to compositions of the present invention, organic solvent, lytic agent, concentrating agents, gellant, softening agent, antioxidant, suspending agent, stabilizing agent, foaming agent (foamingagent), aromatic, surfactant, water, ion-type or nonionic emulsifier, filler, chelating agen, chelating agent, preservative agent, vitamin, blocker, wetting agent, quintessence oil, dyestuff, pigment, hydrophilic or lipophile activating agent, the adjuvant that lipid folliculus or other composition any etc. being usually used in cosmetics are commonly used in cosmeceutical or Dermatology field.Described adjuvant imports with amount normally used in cosmeceutical or Dermatology field.

Further, compositions of the present invention, in order to increase skin improvement effects, can containing promoting the material that skin absorbs.

Detailed description of the invention

Below, structure of the present invention and effect will be further illustrated by test example and dosage form example.But only in order to help to understand the present invention, object illustratively provides for these test examples and dosage form example, and category of the present invention and scope are not limited to following example.

The preparation of [reference example 1] ginsenoside Rf

The ginsenoside Rf used to test effect of the present composition is purchased from rich (AMBO) institute of peace.

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

According to the composition of following table 1, prepare nourishing cream (unit: % by weight) by the method for routine.

Table 1

[test example 1] color improves effect

In order to evaluate cosmetic composition according to the present invention to the effect promoting cutaneous circulation, utilize laser Doppler perfusion imaging instrument (LaserDopplerPeRfusionImager, LDPI; PeriscanPIMII, lark prestige (Perimed) (Stockholm, Sweden)) measure blood circulation degree in skin.Known LDPI measures the sanguimotor instrument in skin, and it is now widely used instrument, be a kind of speed and amount that not only can measure the blood in capillary of skin, but also the very delicate of the flowing in small artery and venule can be measured.

In thermostatic constant wet chamber, after washing one's face with fancy soap, adapt to 30 minutes, and utilize LDPI to determine initial value.First, with LDPI, the initial stage blood flow below the forehead of ice-cold 30 women of trick is at ordinary times measured.Then, make subjects use 1 week described dosage form example 1 and compare dosage form example 1 to measure blood flow afterwards, then the blood flow of mensuration and described initial stage measured value are compared, and its result (dermal blood flow change) is shown in following table 2.

Table 2

Can confirm from the result of above-mentioned table 2, cosmetic composition according to the present invention, compared with the comparison dosage form example 1 not containing ginsenoside Rf, is made dermal blood flow significantly increase, and is promoted by this blood circulation and color is improved.This finally shows can to effectively transmitting skin supplement ingredient, and suppress also delay skin aging to contribute according to the cosmetic composition containing ginsenoside Rf of the present invention.

[test example 2] colour of skin improves effect

In order to the colour of skin understood described dosage form example 1 and compare dosage form example 1 improves effect, 30 tested objects are made to use (evening 1 times/day respectively, smear 1 week altogether) after, FacialStageDM-3 (jasmine spy (Moritex), Japan) the instrument evaluation colour of skin is utilized to improve degree.Adopt the lightness of skin and color change value, and the lightness of skin and color measured value judge to colour of skin improvement rate, and the results are shown in following table 3.Lightness and the higher expression colour of skin of color change value improve.

Table 3

Can confirm from the result of table 3, the comparison dosage form example 1 not containing ginsenoside Rf of the present invention, does not demonstrate the significant colour of skin and improves effect, on the contrary, containing the dosage form example 1 of ginsenoside Rf as effective ingredient, compared with before use, the colour of skin after using is greatly improved.

[test example 3] pore contractive effect

1. by promoting the effect of the pore refining of collagen protein synthesis

Ginsenoside Rf of the present invention is compared the synthesis facilitation effect of collagen protein and transforming growth factor-β (TGF-β) and measures.First, by fibroblast (fibroblast) with every hole 10 ⁵the amount of individual cell is inoculated in 24 holes (well), and carries out cultivating until growing to till about 90%.Used after serum-free cell culture medium (DMEM) cultivates 24 hours, used that 10ng/ml's be dissolved in ginsenoside Rf of the present invention in serum-free medium and TGF-β process respectively, and at CO ₂cultivate 24 hours in incubator.Take out their supernatant, and utilize procollagen type (I) enzyme-linked immunosorbent assay (ELISA) test kit (procollagen type (I) (procollagentype (I); #MK101, TAKARA (will is congratulated, Japan)), whether the increase and decrease of observation procollagen (procollagen) to be, and the results are shown in following table 4.For the synthesis capability of collagen protein, non-process group be set to 100 and contrast.

Table 4

Substances	Collagen protein synthesis ability (%)
		Non-process group	100
TGF-β	183.5
		Ginsenoside Rf	187.6

Can confirm from the result of above-mentioned table 4, ginsenoside Rf of the present invention, compared with positive controls TGF-β, demonstrates the collagen protein synthesis ability of higher levels of excellence.Therefore, can confirm that ginsenoside Rf of the present invention shrinks by the collagen protein growing amount increasing pore periphery the pore broadened.

2. pore contractive effect

In order to understand dosage form example 1 and compare the pore contractive effect of dosage form example 1, evaluate as follows.Selecting men and women's tested object that 20 pore sizes are wide, being divided into two groups by often organizing 10, and according to group smearing dosage form example 1 every day on the face and comparing the nourishing cream of dosage form example 1, smear 4 weeks altogether.The judgement of pore contractive effect is implemented in such a way.Photo before shooting experiment and after 4 weeks, and allow expert be evaluated by naked eyes.The results are shown in (opinion rating: 0-does not shrink completely in following table 5; 5-shrinks a lot).

Table 5

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

Known from the result of above-mentioned table 5, compare the effect that dosage form example 1 does not have pore refining, but dosage form example 1 illustrates the pore contractive effect that can with the naked eye confirm, thus known ginsenoside Rf of the present invention is to the excellent effect of pore refining size.

[test example 4] sebum secretion inhibition

1. by suppressing the effect of the suppression skin supersecretion of 5α-reductase activity

In order to confirm the inhibition to 5α-reductase activity, determine in HEK293-5 α R2 cell [ ¹⁴c] testosterone change into [ ¹⁴c] ratio of dihydrotestosterone (DHT:dihydrotestosterone).On HEK293 cell after transfection p3xFLAG-CMV-5 α R2, and by every hole 2.5 × 10 ⁵the amount of individual cell is inoculated in 24 orifice plates, and carries out cultivating (Parketal., 2003, JDS.Vol.31, pp.191-98).Within second day, change the new culture medium being added with zymolyte and Inhibitors into.By 0.05 μ Ci [ ¹⁴c] testosterone (test kit (AmershamPharmaciabiotech), Britain) is as culture medium substrate.

In order to confirm, to the suppression degree of 5α-reductase activity, to add ginsenoside Rf, and at 37 DEG C, 5%CO ₂cultivate 2 hours in incubator.Now, be used as negative control group by what do not add ginsenoside Rf, the group that will add finasteride (finasteride) is as positive controls.Reclaim culture medium afterwards, and after extracting steroid by 800 μ l ethyl acetate, be separated top organic solvent layer and after drying, remaining residue is dissolved again by 50 μ l ethyl acetate, and on silastometer thin film silica gel 60F254 (Silicaplasticsheetkieselgel60F254), use ethylacetate-hexane (1:1) to launch as solvent.

By plastic sample after being dried in air, shower system is employed in order to measure isotopic amount the sheet plastic of drying and x-ray thin film are together joined bath box in, within 1 week, measure the isotopic mass of testosterone and the dihydrotestosterone stayed on thin film afterwards, then according to following mathematical expression 1 and 2, calculate conversion ratio and obstruction rate respectively, and the results are shown in following table 6.

Mathematical expression 1

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

Mathematical expression 2

Conversion ratio × 100 of obstruction rate (%)=[conversion ratio of the conversion ratio-substances of matched group]/matched group

Table 6

Substances	Conversion ratio (%)	Obstruction rate (%)
			Negative control group	48	-
Positive controls	27.6	42.5
			Ginsenoside Rf	15.4	67.9

Can confirm from the result of above-mentioned table 6, ginsenoside Rf can suppress the activity of 5α-reductase effectively, thus blocking-up testosterone is converted into dihydrotestosterone, and demonstrate inhibition more excellent compared with the finasteride of known suppression 5α-reductase activity.Described 5α-reductase makes testosterone be converted into dihydrotestosterone, thus is combined with intracytoplasmic receptor protein and enters in core, thus activation sebocyte cell promote differentiation, thus make the sebum excessive secretion in sebaceous gland.Therefore, the too much secretion of ginsenoside Rf by effectively suppressing the activity of 5α-reductase to suppress sebum is confirmed.

2. sebum secretion inhibition

In order to understand described dosage form example 1 and compare the sebum secretion inhibition of dosage form example 1, carry out following evaluation.Select 30 and feel men and women's tested object that sebum secretion is many, every day smears dosage form example 1 in the appointed part of facial skin and compares the nourishing cream of dosage form example 1, smears 4 weeks altogether.Sebum is reduced to the judgement of effect, by using sebum tester (SebumeterSM810, C+K Electronics Co., Ltd. (C+KElectronicCo.), Germany) measure the average sebum slip (%) after 2 weeks and 4 weeks respectively, and the results are shown in following table 7.

Table 7

Known from the result of above-mentioned table 7, of the present invention containing ginsenoside Rf as effective ingredient dosage form example 1 with not containing ginsenoside Rf comparison dosage form example 1 compared with, effectively can suppress the sebum of too much secretion.

[dosage form example 2 and compare dosage form example 2 ~ 3]

Prepare dosage form example 2 according to the composition shown in following table 8 and content (% by weight) and compare dosage form example 2 ~ 3.Be described as follows.Dosage form example 2 is the material of mixing ginsenoside Rf, relatively dosage form example 2 is the material not comprising the effective ingredient improving acne skin completely, relatively dosage form example 3 is the standard substance as the benchmark for antimicrbial power, and this material contains the multiplex erythromycin (erythromycin) making acne therapeutic agent.

Dosage form example 2 and to compare the preparation method of dosage form example 2 ~ 3 as described below.The composition of A item in following table 8 is dissolved completely, and in other dissolving tank, the composition of B item is dissolved completely, afterwards B item is added in A item, make it mix solubilized.And wherein, to add the composition of C item according to the mixed proportion recorded in table 8, and after mix homogeneously, filter, thus prepare this compositions.

Table 8

[test example 5] is tested the antimicrbial power of acne bacterium

Use each cosmetic composition prepared according to described dosage form example 2 and the composition comparing dosage form example 2 ~ 3 respectively, the propionibacterium acnes (ATCC6919: culture medium-brain heart infusion broth (BHIbroth)) to as acne pathogenic strain) carry out antimicrbial power test.

As described below to the antimicrbial power test method of acne bacterium.

(1) preparation of test organisms liquid

Propionibacterium acnes is inoculated in and carries out Anaerobic culturel in brain heart infusion broth culture medium and the culture fluid obtained uses as test organisms liquid.

(2) preparation of dilute solution

In the brain heart infusion broth (pH6.8) or LB broth bouillon (pH4.5) of 15ml, add the described test organisms liquid of 0.15ml, and the mixed liquor mixed is used as dilute solution.

(3) preparation of test portion

By from dosage form example 2 and the cosmetic composition stock solution comparing preparation in dosage form example 2 ~ 3, directly use as test portion.

(4) antimicrbial power test

1) add test portion in No. 1, Tissue Culture Plate (96wellplate) row in 96 holes, make to mate with initial concentration, and add dilute solution and make total amount be 200 μ l.

2) by after No. 1 mixed liquor mix homogeneously arranged, the mixed liquor taking out 100 μ l joins in No. 2 rows, and after mix homogeneously, the mixed liquor again taking out 100 μ l joins in No. 3 rows.Carry out doubling dilution (doubledilution) in this way.

3) whether quiescent culture after 24 hours and 48 hours at 32 DEG C, judge the propagation of bacterium by suspension degree, and the Cmin of the propagation not having bacterium be decided to be minimal inhibitory concentration (MinimumInhibitoryConcentration, MIC) value.If the opaque and propagation being difficult to judge bacterium of mixed liquor whether, then confirmed by microscopic examination.

The antimicrbial power result of the test of acne bacterium is shown in following table 9.For minimal inhibitory concentration, be converted into the concentration of the effective ingredient contained in dosage form and labelling.

Table 9

Project	pH	Propionibacterium acnes
			Dosage form example 2	5.7	>41ppm
Relatively dosage form example 2	5.7	Maximum concentration (not having antimicrbial power)
			Relatively dosage form example 3	5.7	>100ppm

Can confirm from the result of above-mentioned table 9, in minimal inhibitory concentration, ppm concentration is less, can think that the antimicrbial power of this material to acne bacterium is effective, use dosage form example 2 time, be used as known acne therapeutic agent erythromycin comparison dosage form example 3 compared with, the ppm concentration demonstrated is significantly low, thus the compositions containing ginsenoside Rf can be confirmed, to test organisms, there is more excellent antimicrbial power.

[test example 6] inhibition test to lipid synthesis (Lipogenesis)

Using the 3T3-L1 cell of the fibroblast (fibroblastcellline) as mice, with 1 × 10 ⁵cells/well is attached at the fetal bovine serum (fetalBovineSerum filled containing 10%, FBS) DMEM (Dulbecco ' smodifiedeagle ' smedium, GIBCOBRL, life technology company) culture medium 6 well culture plates (cultureplate) in.After 2 days, DMEM (FBS containing the 10%) culture medium again more renewed, and cultivate 2 days.Then, with the DMEM (FBS containing 10%) containing 1 μ g/ml insulin (insulin), 0.5mM isobutyl methylxanthine (IBMX) and 0.25 μm of dexamethasone (dexamethasone), again induction is carried out to described cultured cells, and with the ginsenoside Rf of 50 μMs and caffeine process, then, after 2 days, again change the DMEM comprising insulin, and cultivate 5 days.After 5 days, be again replaced with normal incubation medium (DMEM, the FBS containing 10%), and described cell is cultured to described cell is changing into adipose cell from form.

In order to evaluate ginsenoside Rf to the effect suppressing Fat Accumulation in adipose cell, described in utilization, completing the 3T3-L1 adipose cell of differentiation, implementing the Sudan three and dyeing (S4136, Sigma-Aldrich).At normal temperatures, in phosphate buffer, after 4% paraformaldehyde (pH7.2) fixed fat cell, with phosphate buffer (Sphosphatebufferedsaline, PBS) rinse, then, take pictures after dyeing with the Sudan three, and compared by naked eyes.The group of only adding culture medium by not adding substances or compare material uses, as a control group for other comparable group, with the caffeine process of 50 μMs.Fat Accumulation suppresses degree to be by the degree of dyeing being divided into +++, ++ ,+,-, thus give grade, and now, more close +++, represent that dye levels is large.The results are shown in following table 10.

Table 10

Test portion	Obstruction rate %
		Matched group	+++
Comparable group	+
		Ginsenoside Rf	-

Known from above-mentioned table 10, the ginsenoside Rf used in the present invention, not only in adipose cell, the fat mass of accumulation is few, and with hinder the caffeine of material as known lipid synthesis compared with, also has the effect of excellent obstruction lipid synthesis.Therefore, by suppressing lipid synthesis to reduce sebum, thus the generation of acne can be suppressed.

[test example 7] is improved acne and is reduced sebum secretion and have non-stimulated test

Using 30 people suffering from acne as subjects, be divided into three groups often to organize 10, and use by described dosage form example 2 to the subjects corresponding to each group and compare cosmetic composition 1 month prepared by dosage form example 2 ~ 3.Improve standard for acne, be set as 1 point to 5 points, and be marked as 1 and be divided into " not having ", 3 are divided into " common ", and 5 are divided into " very good ".The average mark of 10 is marked in following table 11 as experimental result.

Period is eliminated for acne, to recognize the natural law of elimination as benchmark, for acne recurrence, whether recurred as benchmark after 1 month.For the minimizing of sebum secretion, be set as 1 point to 5 points, and be marked as 1 and be divided into " not having ", 3 are divided into " common ", and 5 are divided into " very good ".The average mark of 10 is marked in following table 11 as experimental result.With (demonstrating the number of irritant reaction)/(overall test number) judge skin irritant with or without.

Table 11

Known from above-mentioned table 11, dosage form example 2 with compare compared with dosage form example 2, acne does not recur, and improves acne on the whole and have excellent effect.In addition, when using the comparison dosage form example 3 containing antimicrbial power standard substance, the effect improving acne is demonstrated, but strong to the stimulation of skin when using, be therefore not suitable for life-time service, but, compositions according to the present invention does not but stimulate, and therefore demonstrates and is also applicable to long-time use.

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

Shampoo is prepared with the composition of following table 12.Particularly, surfactant and glycol distearate are added in Purified Water, and be heated to 80 DEG C, after making its uniform dissolution, under agitation gradually be cooled to 40 DEG C, and, drop into after also mixing according to effective ingredient of the present invention and antiseptic, viscosity modifier, spice and hair conditioner in described mixture, under agitation be cooled to room temperature, thus prepare shampoo.

Table 12

Composition	Dosage form example 4	Relatively dosage form example 5
			Ammonium lauryl sulfate	10	10
Polyoxyethylene lauryl base ammonium sulfate	5	5
			Cocamido propyl betaine	2	2
Glycol distearate	1.5	1.5

Cocomonoethanolamide	0.8	0.8
			Ginsenoside Rf	5.0	-
Polyquaternium-10	0.2	0.2
			Blue No. 1	0.0002	0.0002
Yellow No. 4	0.0001	0.0001
			Methyl parahydroxybenzoate	0.1	0.1
Spice	0.8	0.8
			Citric acid	0.1	0.1
Dimethicone	1.0	1.0
			Water	To 100	To 100

[test example 8] dandruff reduces effect test

The male of 19 to 35 years old that selected 24 dandruff is more, is divided into two groups often to organize 12, and the shampoo using dosage form example 3 with the following methods respectively and compare dosage form example 4 is after 1 month, measures dandruff slip.

Before on-test, usual conventional shampoo cleaning hair, and the accumulation dandruff of two days after gathering hair washing, and by the weight of the dandruff of collection with washed a hair with dosage form example 3 and the shampoo that compares dosage form example 4 every two days respectively and the weight of the dandruff of accumulating two days after off-test compares and evaluates.Now, directly gather the dandruff of accumulation from scalp with vacuum suction apparatus, and obtain dandruff slip according to following equation 3, and the results are shown in following table 13.

Mathematical expression 3

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

Table 13

Known from above-mentioned table 13, when using the dosage form example 3 containing ginsenoside Rf, demonstrate excellent dandruff preventing effectiveness.

[test example 9] prevents the test of the effect of pruritus capitis

Selected 24 men and women of 25 years old to 45 years old more seriously feeling scalp itch, two groups are divided into often to organize 12, and respectively with three days frequency usage dosage form examples 3 once and the shampoo after two weeks that compares dosage form example 4, evaluated preventing the effect of scalp itch by following metewand, and the results are shown in following table 14.

[metewand]

Very excellent-5 points

Excellent-4 points

Generally-3 points

Bad-2 points

Very bad-1 point

Table 14

Classification	Dosage form example 3	Relatively dosage form example 4
			The removal effect of the pruritus of scalp	4.5	2.3

Known from above-mentioned table 14, when using the dosage form example 3 containing ginsenoside Rf, demonstrate more excellent effect to preventing scalp itch.

[test example 10] increases the effect assessment of potassium ion channel activity

Minoxidil as alopeciaing therapeutic agent is known as potential mitochondrion K ~+Channel Opener (K _aTPchannelopener), be the representative drugs used in the treatment of androgenetic alopecia.In order to evaluate the mechanism of this minoxidil, employ following test method(s).Described test method(s) is use in the fibroblast of composition scalp corium to stop K _aTPthe tolbutamide (SIGMAAIDRICH, T0891) of passage processes, thus antiproliferative effect, and again open potassium-channel and recover cell proliferation.

In order to evaluate the K as this compositions _aTPthe function of channel opener, present invention uses mouse embryo fibroblasts system (Mouseembryonicfibroblastcellline, the NIH3T3) cell line as fibroblast.This cell is use 3T3 scheme, the cell line obtained carrying out nature immortalization from the middle fibroblast be separated of NIH Swiss mouse embryo (Swissmouseembryo).By described cell line in the DMEM (GibcoBRL, Gaithersburg, MD, the U.S.) containing 10%FBS, at maintenance 5%CO ₂, cultivate 24 hours in the incubator of 37 DEG C.NIH3T3 is inoculated in 96 orifice plates, and cultivate in the incubator of 37 DEG C after 24 hours, process with the tolbutamide of 2.5mM, and after 10 minutes, be used separately as the minoxidil of 10 μMs of positive controls and the ginsenoside Rf of 2.5ppm, 5ppm and 10ppm concentration process, and through 48 hours after drug treating, then use WST-1 test kit (Roche (Roche)) to measure ability of cell proliferation.And the results are shown in following table 15.

Table 15

Classification	Ability of cell proliferation (%)
		Non-processor matched group (contrast)	100
Minoxidil	132
		Ginsenoside Rf (2.5ppm)	115
Ginsenoside Rf (5ppm)	121
		Ginsenoside Rf (10ppm)	132

Known from above-mentioned table 15, when processing with ginsenoside Rf, fibroblastic propagation is restored, and ability of cell proliferation depends on the concentration of the ginsenoside Rf of process and increases, and when can confirm to process with the ginsenoside Rf of 10ppm, cell proliferation returns to by level during minoxidil process.

The melanin of [test example 11] ginsenoside Rf generates facilitation effect test

Add in RPMI culture medium the fetal bovine serum of 5%, the benzylpenicillin of 100IU and 0.2 μM p-phthalic acid (TPA) culture medium in, by melanocyte with 50,000 cells/well is inoculated in 24 orifice plates.Second day, to the cell of inoculation, process with the ginsenoside Rf as substances of the ultimate density of 10ppm or 50ppm.Using by the group of DMSO process of 0.1% as group as negative control group, using by the group of the IBMX process of 100 μMs as positive controls, above-mentioned each group is cultivated three days at 37 DEG C of temperature.After cultivation, with phosphate buffer (PBS) cleaning orifice, and add the 1NNaOH of 100 μ l in every hole after, the melanin in dissolved cell.Utilize slat chain conveyor analyzer (microplatereader), measure under 405nm by the melanic absorbance (cooperative effect 2, microplate reader (VT, the U.S.)) dissolved.The result that the effect and matched group that promote the melanic generation of ginsenoside Rh compare is shown in following table 16.

Table 16

Test portion	B16 cell amount (%)
		DMSO(0.1％)	100
IBMX(100μM)	120
		Ginsenoside Rf (10ppm)	112
Ginsenoside Rf (50ppm)	123

Known from above-mentioned table 16, ginsenoside Rf promotes melanocytic B16 cell, thus increases melanic generation, therefore, it is possible to demonstrate the effect of excellent promotion melanin generation.

[test example 12] ginsenoside Rf promotes the effect that transcription factor (MITF) and tryrosinase are expressed in melanocyte

Utilize 501mel cell line, with 500,000 cells/well is inoculated in 6 orifice plates, and in each hole, dimethyl sulfoxide (DMSO) with 0.1% process as positive controls, with the IBMX process of 100 μMs as positive controls, and with 10ppm life saponin Rf process as experimental group, and cultivate after 24 hours, 48 hours and 72 hours at 37 DEG C of temperature, obtain protein.For the protein so obtained, MITF and tryrosinase antibody is utilized to carry out the test of Western blot (WesternBlot) method.Protein Extraction and Western blot are implemented by the normally 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 compares with this value and be shown in following table 17.

Table 17

Can confirm from above-mentioned table 17, ginsenoside Rf improves the expression of MITF and tyrosinase protein matter in melanocyte.

The antimicrbial power evaluation of [test example 13] ginsenoside Rf

In order to evaluate the antimicrbial power of ginsenoside Rf, implement antibacterial experiment.Concrete test method is as follows.

The staphylococcus aureus (Staphylococcusaureus) used in experiment, escherichia coli (Escherichiacoli) and Pseudomonas aeruginosa (Pseudomonasaeruginosa) cultivate in trypticase soya broth culture medium (TrypticSoyBroth); Candida albicans (Candidaalbicans) and aspergillus niger (Aspergillusniger) cultivate in Sabouraud dextrose broth bouillon (SabouraudDextroseBroth).The diluent that culture fluid carries out diluting with 1/100 (albicans strain is using 1/10) in each culture medium is used as test organisms liquid.For aspergillus niger, will 2 × 10 be prepared into ⁸the spore suspension of cfu/ml uses as test organisms liquid.

The test organisms liquid of 0.15ml is added and the mixed liquor mixed uses as dilute solution in each culture medium of 15ml.

In No. 1,96 orifice plate row, add 10ppm ginsenoside Rf respectively with the amount of every hole 16 μ l, and add dilute solution with every hole 184 μ l amount.Dilute solution is added in the amount of other Zhong Yimei hole 100, hole μ l.After No. 1 mixed liquor mix homogeneously arranged, the mixed liquor taking out 100 μ l joins in No. 2 rows, and after mix homogeneously, the mixed liquor again taking out 100 μ l joins in No. 3 rows.Carry out doubling dilution in this way.

Staphylococcus aureus, escherichia coli and Pseudomonas aeruginosa cultivate in the temperature chamber of 32 DEG C; Candida albicans and aspergillus niger cultivate in the temperature chamber of 25 DEG C.

After 48 hours, whether confirm the increment of bacterium by suspensibility and microscope, thus determine minimal inhibitory concentration (MIC) value, and the results are shown in following table 18.

Table 18

As shown in above-mentioned table 18, can confirm that ginsenoside Rf demonstrates antimicrbial power to multiple bacterial strain, and can predict that ginsenoside Rf can be worked as natural antiseptic agent or antibacterial in compositions by these.

Claims

1. a Dermatologic preparation composition, is characterized in that, described Dermatologic preparation composition contains ginsenoside Rf as effective ingredient.

2. Dermatologic preparation composition according to claim 1, is characterized in that, described ginsenoside Rf represents with following chemical formula 1:

3. Dermatologic preparation composition according to claim 1 and 2, is characterized in that, in composition total weight, described Dermatologic preparation composition contains the described ginsenoside Rf of 0.001 ~ 50 % by weight.

4. Dermatologic preparation composition according to claim 1 and 2, is characterized in that, described compositions is for improving acne.

5. Dermatologic preparation composition according to claim 1 and 2, is characterized in that, described compositions is used for antibacterial.

6. Dermatologic preparation composition according to claim 1 and 2, is characterized in that, described compositions is for improving color and the colour of skin.

7. Dermatologic preparation composition according to claim 1 and 2, is characterized in that, described compositions is used for pore refining.

8. Dermatologic preparation composition according to claim 1 and 2, is characterized in that, described compositions is for regulating sebum.

9. Dermatologic preparation composition according to claim 1 and 2, is characterized in that, described compositions is for improving skin problem.

10. Dermatologic preparation composition according to claim 1 and 2, is characterized in that, described compositions is for promoting hair growth.

11. Dermatologic preparation compositions according to claim 1 and 2, is characterized in that, described compositions is used for preventing poliosis.

12. Dermatologic preparation compositions according to claim 1 and 2, is characterized in that, described compositions is used for anti-dandruff.

13. Dermatologic preparation compositions according to claim 1 and 2, is characterized in that, described compositions uses as natural antiseptic agent.

14. 1 kinds are being improved the application in acne containing ginsenoside Rf as the Dermatologic preparation composition of effective ingredient.

15. 1 kinds containing the Dermatologic preparation composition application in antibacterial of ginsenoside Rf as effective ingredient.

16. 1 kinds are being improved the application in color and the colour of skin containing ginsenoside Rf as the Dermatologic preparation composition of effective ingredient.

17. 1 kinds containing the Dermatologic preparation composition application in pore refining of ginsenoside Rf as effective ingredient.

18. 1 kinds containing the Dermatologic preparation composition application in adjustment sebum of ginsenoside Rf as effective ingredient.

19. 1 kinds are being improved the application in skin problem containing ginsenoside Rf as the Dermatologic preparation composition of effective ingredient.

20. 1 kinds containing the Dermatologic preparation composition application in promotion hair growth of ginsenoside Rf as effective ingredient.

21. 1 kinds are preventing the application in poliosis containing ginsenoside Rf as the Dermatologic preparation composition of effective ingredient.

22. 1 kinds containing the Dermatologic preparation composition application in anti-dandruff of ginsenoside Rf as effective ingredient.

23. 1 kinds containing the Dermatologic preparation composition application when as natural antiseptic agent use of ginsenoside Rf as effective ingredient.