CN105246489B - Dermatologic preparation composition containing ginsenoside RF - Google Patents

Dermatologic preparation composition containing ginsenoside RF Download PDF

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CN105246489B
CN105246489B CN201480029495.9A CN201480029495A CN105246489B CN 105246489 B CN105246489 B CN 105246489B CN 201480029495 A CN201480029495 A CN 201480029495A CN 105246489 B CN105246489 B CN 105246489B
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ginsenoside
effect
skin
dosage form
composition
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CN105246489A (en
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金东泫
柳权烈
李沃澯
廉明勋
曺濬喆
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Aimeri Pacific
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Aimeri Pacific
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Priority to CN201711252406.9A priority patent/CN107898656B/en
Priority to CN201810479801.9A priority patent/CN108498365B/en
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
    • A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/63—Steroids; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0014—Skin, i.e. galenical aspects of topical compositions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/10—Anti-acne agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/02—Local antiseptics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/008—Preparations for oily skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q5/00—Preparations for care of the hair
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q5/00—Preparations for care of the hair
    • A61Q5/006—Antidandruff preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q5/00—Preparations for care of the hair
    • A61Q5/10—Preparations for permanently dyeing the hair
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q7/00—Preparations for affecting hair growth
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
    • A61K2800/52—Stabilizers
    • A61K2800/524—Preservatives

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Abstract

Contain composition of the ginsenoside Rf as effective component the present invention relates to a kind of.The composition is capable of providing the effect of the effect, convergence skin and pore refining that improve acne and skin problem, and is capable of providing the effect for improving skin color, promotion hair growth, improves white hair, anti-dandruff and anti-corrosion effect.

Description

Dermatologic preparation composition containing ginsenoside RF
Technical field
The present invention relates to a kind of composition, the composition contains ginsenoside Rf (Ginsenoside Rf), so as to It is enough that the effect of the effect, convergence skin and pore refining that improve acne and skin problem is provided, and it is capable of providing improvement skin The effect of color promotes hair growth, improves white hair, anti-dandruff and anti-corrosion effect.
Background technique
First defensive barrier of the skin as human body, have so that intracorporeal organ from such as temperature and humidity variation, The stimulation of the external environment of ultraviolet light, public hazards substance and the function of protecting intracorporeal organ.With advancing age, skin will be due to Inherent, external factor and change.That is, the various hormone secretions due to adjusting metabolism will subtract in terms of the inherence It is few, and the function of immunocyte and cell activity will reduce, therefore the immune protein of needed by human body and organism constitutive protein Biosynthesis will reduce.For in terms of external, due to the destruction of ozone layer, the ultraviolet light of earth's surface is reached from sunray Content will increase, and as the in-depth of environmental pollution, free radical and active oxygen will increase, thus not only make the thickness of skin Reduce, wrinkle increases, elastic force reduces, and keeps skin complexion dimmed, problem (trouble) always occurs for skin, and make mole and Freckle and senile plaque also increase, so as to 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 generate skin condition variation, and maintain health skin Skin state, people make great efforts always by adding the physiological activator obtained from existing various animals, plant, microorganism etc. Enforcement of going forward side by side is added in cosmetics to be used to improve skin condition.
Ginsenoside Rf is included in one kind of the saponin in ginseng, it is known that shows anti-lipid peroxidation effect and prevents Imperial physiological activity by the effect of alcohol-induced cerebral dysgenesis etc., inhibit voltage-dependent ca channel (Nah, et al., Proc.Natl.Acad.Sci.USA, 92,8739-8743,1995) and anti-pain effect (Shin et al., submited, 1997).But to contain ginsenoside Rf as the composition of effective component improvement acne and skin problem effect, convergence The effect of skin and pore refining, the effect for improving skin color promote hair growth, improve white hair, anti-dandruff and anti-corrosion Effect is not yet reported that.
Summary of the invention
Technical problems to be solved
In this regard, the inventors discovered that ginsenoside Rf is capable of providing the effect for improving acne and skin problem, convergence skin And the effect of pore refining, and be capable of providing the effect for improving skin color, promotion hair growth, improve white hair, anti-scalp Bits and anti-corrosion effect, so as to complete the present invention.
Therefore, the object of the present invention is to provide a kind of Dermatologic preparation composition, the composition contains ginsenoside Rf so as to provide the effect of the effect for improving acne and skin problem, convergence skin and pore refining, and shows and changes The effect of kind skin color promotes hair growth, improves white hair, anti-dandruff and anti-corrosion effect.
Technical solution
To achieve the goals above, the present invention, which provides, a kind of contains ginsenoside Rf as effective component for improving Cuo The Dermatologic preparation composition of sore.
In addition, the present invention provides a kind of skin for being used to improve color and the colour of skin for containing ginsenoside Rf as effective component 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 Rf as effective component Agent composition.
In addition, the present invention provides and a kind of contains ginsenoside Rf as effective component for trichogenous skin 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 Rf as effective component Agent composition.
In addition, the present invention provides a kind of external preparation for skin for anti-dandruff for containing ginsenoside Rf as effective component Agent composition.
In addition, the present invention provides a kind of skin used as natural antiseptic agent for containing ginsenoside Rf as effective component Skin preparation composition for external use.
Beneficial effect
Composition of the invention contains ginsenoside Rf, so as to provide effect that is anti-inflammatory, improving acne and skin problem Fruit, the effect for restraining skin and pore refining, and be capable of providing the effect for improving skin color, promotion hair growth, improve White hair, anti-dandruff and anti-corrosion effect.
Preferred forms
Composition according to the present invention contains ginsenoside Rf as effective component.
Ginsenoside Rf used in the present invention has the structure of following chemical formula 1:
Chemical formula 1
Ginsenoside Rf of the invention can be extracted from plants, and can also be synthesized according to method as known in the art And use, the ginsenoside Rf of mercantile-type sale also can be used.In addition, ginsenoside Rf can obtain from ginseng extract .At this moment the type of the ginseng used is not particularly limited, and water ginseng, red ginseng, white ginseng, Tai Ji ginseng and tail ginseng etc. can be used.And And the ginseng extract not only include as extract, decoct in ginseng mention obtained from leachate, also comprising to the part of leachate Or concentrate obtained from being entirely concentrated again, or impregnating, decoction, piece dry again to the concentrate and prepare AgentExtracts are flowed, and play the chemical substance of main effect included in ginseng, but also include plant Itself, and the extract in all parts of ginsengs such as stem, root, leaf, flower, fruit can be used, a certain spy can't be defined in Determine the extract of part.In addition, well known method can be used in the method for extracting ginsenoside Rf from ginseng extract.
Specifically, the ginsenoside Rf can be made by method as known in the art using water or organic solvent After standby ginseng extract, therefrom it is isolated.Organic solvent used in the present invention can selected from ethyl alcohol, methanol, butanol, The mixed solvent of ether, ethyl acetate, chloroform and 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 hours.If exceeding the Extracting temperature and the range of extraction time, Extraction efficiency can reduce, or the variation of ingredient can occur.
Composition of the invention preferably comprises the ginsenoside that 0.001 to 50 weight % is calculated as with composition total weight Rf.This is because there is the effect of being brought by the ingredient and effect when the content of the effective component is less than 0.001 weight % The faint problem of fruit；When more than 50 weight %, there is a problem of in cutaneous safety or dosage form.
The Dermatologic preparation composition that composition of the invention can be used as improving acne to use antibacterial effect It is excellent, it is especially excellent to the antibacterial effect of acne pathogenic bacteria, and antiphlogistic effects are provided.
The Dermatologic preparation composition that composition of the invention can be used as improving color and the colour of skin comes using by it When being used in skin, by expanding capillary, and promotes blood circulation and come smoothly by nutrition supply to skin, and inhibit skin Aging, therefore, the effect for improving color and the colour of skin are brilliant.
Composition of the invention can be used as pore refining, adjust sebum and improve the skin preparations for extenal use of skin problem Composition comes using when being used in skin, inhibiting the sebum of excessive secretion, and elimination and collagen by promoting active oxygen The synthesis of albumen carrys out pore refining, and due to the reduction of inflammatory factor expression, inhibit the effect of skin problem brilliant.
Composition of the invention, which can be used as, to be come for trichogenous composition using by promoting stand-down It is converted into the anagen hair period hair cycle, to promote the growth of hair, and not only promotes the generation of new hair, but also Existing hair health is grown, and prevention is provided and inhibits the phenomenon that hair falls off from scalp, hypotrichosis or attenuates State effect.
Composition of the invention can be used as preventing the composition of white hair from coming using by increasing in melanocyte The expression of transcription factor (MITF) carry out activation of melanocyte, and promote the synthesis of melanin, to provide advance preventing dialogue The induction of hair and the effect for promoting to induce dark hair.
Composition of the invention can be used as preventing the Dermatologic preparation composition of dandruff from coming using by having Effect discharge is accumulated in the toxin of hair and scalp to purify scalp, and proliferation and growth by inhibiting dandruff bacterium can be pre- Anti- scalp inflammation reaction, also, since the anti-oxidation function of generation and the effect of inhibitory activity oxygen is brilliant, it is capable of providing It calms and strengthens scalp, and strengthen the effect of intrinsic phylactic power defensive power.
Composition of the invention, which can be used as natural antiseptic agent composition, to be come using since composition of the invention is natural Ingredient, therefore anti-corrosion effect brilliance and harmless effect are provided.
Composition according to the present invention can the agent containing acceptable carrier or substrate on cosmeceutical or Dermatology Type.Its whole dosage form that can be used as suitable local use provides, for example, can be provided as solution, gel, solid, paste The anhydrous product of shape The dispersed oil phase in water phase and obtain lotion, suspension, microemulsion, The form of microcapsules, subparticle ball or ionic (liposome) and non-ionic folliculus dispersing agent, or frost, toner, wash Agent, powder, ointment, spray or concealing stick form.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, Dermatologic preparation composition of the invention is white for preventing dandruff, for hair growth or for preventing When hair, the composition for scalp and hair can be used as and dosage form, dosage form are not particularly limited, such as can be by dosage form For hair oil, trichotrophy toner, scalp care agent, hair-care agent, shampoo, hair conditioner, hair lotion or scalp hair Send out dual-purpose care agent etc..
Also, composition according to the present invention may 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 lipophilic 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.
Also, composition of the invention can contain the substance for promoting skin to absorb to increase skin improvement effects.
Specific embodiment
Hereinafter, structure and effect of the invention will be further illustrated by test example and dosage form example.However, these are tested Example and dosage form example are provided only to assist in the present invention is understood as the purpose of illustration, and scope of the invention and range are simultaneously It is not limited to following examples.
The preparation of [reference example 1] ginsenoside Rf
The ginsenoside Rf used to be tested to the effect of present composition studies purchased from rich (AMBO) is pacified Institute.
[dosage form example 1 and compare dosage form example 1]
According to the composition of following table 1, nourishing cream (unit: weight %) is prepared by conventional method.
Table 1
[test example 1] color improvement
It is how general using laser in order to evaluate the effect that cosmetic composition according to the present invention recycles promotion skin blood Strangle blood flow imaging instrument (Laser Doppler Perfusion Imager, LDPI；Periscan PIM II, lark prestige (Perimed) (Stockholm, Sweden)) measurement skin in blood circulation degree.Known LDPI is the blood measured in skin The instrument of liquid circulation, and it is now widely used instrument, is a kind of blood that can not only measure in capillary of skin The speed and amount of liquid, but also 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 determine initial value using LDPI.Firstly, It is determined with the initial stage blood flow below the forehead of 30 LDPI ice-cold to usually trick women.Then, make test pair Blood flow is measured later as using 1 week dosage form example 1 and comparing dosage form example 1, then by the blood flow of measurement and the initial stage Measured value is compared, and its result (skin blood flow variation) is shown in following table 2.
Table 2
From the result of above-mentioned table 2 it has been confirmed that cosmetic composition according to the present invention with do not contain ginsenoside Rf Comparison dosage form example 1 compare so that skin blood flow dramatically increases, and promote to change color by this blood circulation It is kind.This finally show the cosmetic composition according to the present invention containing ginsenoside Rf can to effective transmitting skin-nourishing at Point, and simultaneously delay skin aging contributes for inhibition.
[test example 2] 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, use 30 test objects 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.The colour of skin is improved using the lightness and color change value of skin and the lightness of skin and color measured value Rate is judged, and the results are shown in following Table 3.Lightness and the higher expression colour of skin of color change value are improved.
Table 3
It has been confirmed that not containing the comparison dosage form example 1 of ginsenoside Rf of the invention from the result of table 3, do not show The significant colour of skin improves effect, on the contrary, contain dosage form example 1 of the ginsenoside Rf as effective component, before use compared with, use The colour of skin afterwards is greatly improved.
[test example 3] pore contractive effect
1. the effect of the pore refining by promoting collagen synthesis
By ginsenoside Rf of the invention to the synthesis facilitation effect of collagen and transforming growth factor-β (TGF-β) into It goes relatively and is determined.Firstly, by fibroblast (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 the ginsenoside Rf and TGF-β of the invention of 10ng/ml being dissolved in serum free medium at Reason, and in CO2It is cultivated 24 hours 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, TAK ARA (congratulate, day by will This)), whether observing the increase and decrease of procollagen (procollagen), and it the results are shown in following table 4.For collagen Synthesis capability, non-process group is set as 100 and is compared.
Table 4
Substances Collagen synthesis ability (%)
Non-process group 100
TGF-β 183.5
Ginsenoside Rf 187.6
It has been confirmed that ginsenoside Rf of the invention is compared with positive controls TGF-β from the result of above-mentioned table 4, show Higher levels of excellent collagen synthesis ability is shown.Therefore, it can be confirmed that ginsenoside Rf of the invention passes through increase The collagen production quantity on pore periphery shrinks the pore to broaden.
2. pore contractive effect
In order to understand dosage form example 1 and compare the pore contractive 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 daily according to group on the face and comparing The nourishing cream of dosage form example 1 is smeared 4 weeks altogether.The judgement of pore contractive effect is implemented in the following way.Shooting experiment before and 4 weeks Photo afterwards, and expert is allowed to evaluate by naked eyes.The results are shown in (opinion rating: 0- is absolutely not received in following table 5 Contracting；5- shrinks very much).
Table 5
Substances Opinion rating
Dosage form example 1 4
Compare dosage form example 1 0
It is found that comparing the effect of the not no pore refining of dosage form example 1 from the result of above-mentioned table 5, however dosage form example 1 shows energy The pore contractive effect of enough naked eyes confirmations, to know that ginsenoside Rf of the invention is excellent to the effect of pore refining size It is different.
[test example 4] sebum secretion inhibitory effect
1. by inhibiting the active effect for inhibiting skin hypersecretion of 5α-reductase
In order to confirm to the active inhibitory effect of 5α-reductase, determined in HEK293-5 α R2 cell [14C] testosterone turn Chemical conversion [14C] dihydrotestosterone (DHT:dihydrotestosterone) ratio.P3x FLAG- is transfected on HEK293 cell After CMV-5 α R2, and press every 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 Inhibitors into.By 0.05 μ Ci [14C] testosterone (kit (Amersham Pharmacia biotech), Britain) be used as culture substrate.
In order to confirm to the active inhibition level of 5α-reductase, it is added ginsenoside Rf, and at 37 DEG C, 5%CO2Culture It is cultivated 2 hours in device.At this point, no addition ginsenoside Rf is used as negative control group, Finasteride will be added (finasteride) group is used as positive controls.Recycle culture medium later, and with 800 μ l ethyl acetate extract steroids it Afterwards, the organic solvent layer on top is separated after drying, is redissolved remaining residue with 50 μ l ethyl acetate, and in silastomer On 60 F254 of film silica gel (Silica plastic sheet kieselgel60 F254), using ethylacetate-hexane (1: 1) it is unfolded as solvent.
After being dried in air by plastic sample, shower system has been used in order to measure the amount of isotopeDry sheet plastic and x-ray film are added to bath box together In, measurement stays in the isotopic mass of testosterone and dihydrotestosterone on film after 1 week, then according to following mathematical expressions 1 and 2, respectively Conversion ratio and obstruction rate are calculated, and the results are shown in following table 6.
Mathematical expression 1
Radiant/total radiant × 100 in conversion ratio (%)=region DHT
Mathematical expression 2
Obstruction rate (%)=[conversion ratio-substances conversion ratio of control group]/control group conversion ratio × 100
Table 6
Substances Conversion ratio (%) Obstruction rate (%)
Negative control group 48 -
Positive controls 27.6 42.5
Ginsenoside Rf 15.4 67.9
It has been confirmed that ginsenoside Rf can effectively inhibit the activity of 5α-reductase from the result of above-mentioned table 6, from And testosterone is blocked to be converted into dihydrotestosterone, and show compared with the active Finasteride of known inhibition 5α-reductase more Add excellent inhibitory effect.The 5α-reductase makes testosterone be converted into dihydrotestosterone, thus with intracytoplasmic receptor protein knot It closes and enters in core, to activate sebocyte cell and promote to break up, to make the sebum excessive secretion in sebaceous glands.Cause This, it is thus identified that ginsenoside Rf inhibits the excessive secretion of sebum by effectively inhibiting the activity of 5α-reductase.
2. sebum secretion inhibitory effect
In order to understand the dosage form example 1 and compare the sebum secretion inhibitory effect of dosage form example 1, following evaluation has been carried out.Choosing 30 men and women's test objects felt more than sebum secretion out are smeared dosage form example 1 in the appointed part of facial skin daily and are compared The nourishing cream of dosage form example 1 is smeared 4 weeks altogether.The judgement that effect is reduced for sebum, by using sebum tester (Sebumeter SM810, C+K Electronics Co., Ltd. (C+K Electronic Co.), Germany) 2 weeks and 4 are passed through in measurement respectively Average sebum slip (%) after week, and the results are shown in following table 7.
Table 7
From the result of above-mentioned table 7 it is found that it is of the invention contain ginsenoside Rf as effective component dosage form example 1 with not Comparison dosage form example 1 containing ginsenoside Rf is compared, and the sebum excessively secreted can be effectively inhibited.
[dosage form example 2 and compare dosage form example 2~3]
Dosage form example 2 is prepared according to the ingredient and content (weight %) that show in following table 8 and compares dosage form example 2~3.Tool Body is described as follows.Dosage form example 2 is the substance for mixing ginsenoside Rf, and comparing dosage form example 2 is absolutely not comprising improving acne skin The substance of the effective component of skin, comparing dosage form example 3 is the standard substance as the benchmark for antimicrbial power, which contains multi-purpose Make the erythromycin (erythromycin) of acne therapeutic agent.
Dosage form example 2 and the preparation method for comparing dosage form example 2~3 are as described below.A in following table 8 ingredients is completely molten Solution, and in other dissolving tanks, B ingredients are completely dissolved, B are added in A later, keep its mixing solvable Change.And wherein, to add C ingredients according to the mixed proportion recorded in table 8, and after mixing, it is filtered, from And prepare this composition.
Table 8
[test example 5] tests the antimicrbial power of acne bacterium
It is used respectively according to the dosage form example 2 and compares the composition of dosage form example 2~3 and each cosmetic composition for preparing, To the propionibacterium acnes (ATCC6919: culture medium-brain heart infusion broth (BHI broth)) as acne pathogenic strain) into The test of row antimicrbial power.
It is as described below to the antimicrbial power test method of acne bacterium.
(1) preparation of test bacteria liquid
Propionibacterium acnes are inoculated in brain heart infusion broth culture medium and carry out the work of culture solution obtained from Anaerobic culturel For test bacteria liquid use.
(2) preparation of dilute solution
The examination of addition 0.15ml in the brain heart infusion broth (pH6.8) or LB broth bouillon (pH4.5) of 15ml Test bacterium solution, and using the mixed liquor mixed as dilute solution come using.
(3) preparation of sample
From dosage form example 2 and the cosmetic composition stoste prepared in dosage form example 2~3 will be compared, made directly as sample With.
(4) antimicrbial power is tested
1) sample is added in tissue culture plate (96 well plate) 1 row in 96 holes, so that with initial concentration Match, and dilute solution is added to make 200 μ l of total amount.
2) after mixing, the mixed liquor for taking out 100 μ l is added in No. 2 rows the mixed liquor for arranging No. 1, and is uniformly mixed Afterwards, the mixed liquor for taking out 100 μ l again is added in No. 3 rows.Doubling dilution (double is carried out in this way dilution)。
3) whether judging the proliferation of bacterium with suspension degree, and will not have after stationary culture 24 hours and 48 hours at 32 DEG C There is the Cmin of the proliferation of bacterium to be set to minimum inhibitory concentration (Minimum Inhibitory Concentration, MIC) value. Whether being difficult to judge the proliferation of bacterium if mixed liquor is opaque, confirmed by micro- sem observation.
The antimicrbial power test result of acne bacterium is shown in following table 9.For minimum inhibitory concentration, it is converted into dosage form and contains The concentration of some effective component and mark.
Table 9
Project pH Propionibacterium acnes
Dosage form example 2 5.7 >41ppm
Compare dosage form example 2 5.7 Maximum concentration (does not have antimicrbial power)
Compare dosage form example 3 5.7 >100ppm
From the result of above-mentioned table 9 it has been confirmed that in minimum inhibitory concentration, ppm concentration is smaller, it is believed that the substance It is effective to the antimicrbial power of acne bacterium, when using dosage form example 2, the agent compared with used as the erythromycin of well known acne therapeutic agent Type example 3 is compared, and the ppm concentration shown is significantly low, whereby it was confirmed that the composition containing ginsenoside Rf, has test organisms There is superior antimicrbial power.
[test example 6] tests the inhibition of lipid synthesis (Lipogenesis)
By the 3T3-L1 cell as the fibroblast of mouse (fibroblast cell line), with 1 × 105Carefully Born of the same parents/hole is attached at the DMEM (Dulbecco ' s for filling the fetal bovine serum (fetal Bovine Serum, FBS) containing 10% Modified eagle ' s medium, GI BCO BRL, live technology company) culture medium 6 well culture plate (culture Plate in).After 2 days, new DMEM (FBS containing 10%) culture medium is re-replaced, and cultivate 2 days.Then, with containing There are 1 μ g/ml insulin (insulin), 0.5mM isobutyl methylxanthine (IBMX) and 0.25 μm of dexamethasone (dexamethasone) DMEM (FBS containing 10%) carries out induction to the cell of the culture again, and with 50 μ Then the ginsenoside Rf of M and caffeine processing after 2 days, re-replace the DMEM comprising insulin, and cultivate 5 days.5 After it, normal incubation medium (DMEM, the FBS containing 10%) is re-replaced into, and cultivated the cell to the cell Change lipoblast from form.
In order to evaluate the effect of ginsenoside Rf is to Fat Accumulation in fat cell is inhibited, the completion differentiation is utilized 3T3-L1 fat cell implements the Sudan three and dyes (S4136, Sigma-Aldrich).At normal temperature, in phosphate buffer In, after 4% paraformaldehyde (pH7.2) fixed fat cell, with phosphate buffer (phosphate buffered Saline, PBS) it is rinsed, then, photo is shot after being dyed with the Sudan three, and be compared by naked eyes.To not have Addition substances or the group for comparing substance and only adding culture medium use as a control group, for other comparative groups, with 50 μ The caffeine of M is handled.Fat Accumulation inhibition level is by the way that the degree of dyeing to be divided into +++, ++ ,+,-, so that grade is assigned, At this point, closer +++, indicate that dye levels are big.The results are shown in following table 10.
Table 10
Sample Obstruction rate %
Control group +++
Comparative group +
Ginsenoside Rf -
It is found that ginsenoside Rf used in the present invention, the fat mass not only accumulated in fat cell from above-mentioned table 10 It is few, and compared with the caffeine for hindering substance as well known lipid synthesis, also with the excellent effect for hindering lipid synthesis Fruit.Therefore, sebum is reduced by inhibiting lipid synthesis, so as to inhibit the generation of acne.
[test example 7] improves acne and reduces sebum secretion and have non-stimulated test
Using 30 people with acne as subjects, it is divided into three groups, and the examination to each group is corresponded to every group 10 Object is tested using by the dosage form example 2 and comparing dosage form example 2~3 prepare cosmetic composition 1 month.Acne is changed Kind standard, is set as 1 point to 5 points, and is marked as 1 point as " not having ", and 3 points are " common ", and 5 points are " very good ".By 10 put down Equal score is marked in following table 11 as experimental result.
Period is eliminated for acne, to recognize the number of days of elimination as benchmark, acne is recurred, to answer after 1 month As benchmark whether hair.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 ".It is marked on 10 average marks as experimental result in following table 11.With (showing stimulation The number of reaction)/(overall test number) judge the presence or absence of skin irritatin.
Table 11
It is found that the dosage form example 2 compared with of dosage form example 2 is compared from above-mentioned table 11, acne does not recur, and on the whole to Cuo Sore improvement has the effect of excellent.In addition, showing improves Cuo when using comparison dosage form example 3 containing antimicrbial power standard substance The effect of sore, but it is strong to the stimulation of skin when use, therefore be not suitable for being used for a long time, however, composition according to the present invention But it does not stimulate, thus it is shown that being also suitble to use for a long time.
[dosage form example 3 and compare dosage form example 4]
Shampoo is prepared with the composition of following table 12.Specifically, surfactant and glycol distearate are added It is added in Purified Water, and is heated to 80 DEG C, after making its uniform dissolution, be cooled to 40 DEG C gradually under stiring, also, described mixed It closes after putting into effective component according to the present invention and preservative, viscosity modifier, fragrance and hair conditioner in object and mixing, In It is cooled to room temperature under stirring, to prepare shampoo.
Table 12
[test example 8] dandruff reduces effect test
19 to 35 years old more males of selected 24 dandruffs, are divided into two groups with every group 12, and divide with the following methods Not using dosage form example 3 and after comparing shampoo 1 month of dosage form example 4, dandruff slip is measured.
Before on-test, hair usually is cleaned with conventional shampoo, and acquires the dandruff for accumulating two days after hair washing, And by the weight of the dandruff of acquisition with respectively with dosage form example 3 and compared with dosage form example 4 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, direct from scalp with vacuum suck device The dandruff of accumulation is acquired, and finds out dandruff slip according to following the equation 3, and the results are shown in following table 13 In.
Mathematical expression 3
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 13
It is found that showing that excellent dandruff prevents when using dosage form example 3 containing ginsenoside Rf from above-mentioned table 13 Effect.
[test example 9] prevents the test of the effect of pruritus of scalp
Selected 24 are divided into two groups than more serious 25 years old to 45 years old men and women for feeling itching of the scalp with every group 12, And respectively with frequency usage dosage form example 3 once three days and the shampoo for comparing dosage form example 4 after two weeks, pass through following evaluation criteria The effect for preventing itching of the scalp is evaluated, and the results are shown in following table 14.
[evaluation criteria]
It is -5 points very excellent
Excellent -4 points
Generally -3 points
Bad -2 points
It is -1 point very bad
Table 14
Classification Dosage form example 3 Compare dosage form example 4
The removal effect of the pruritus of scalp 4.5 2.3
From above-mentioned table 14 it is found that when using dosage form example 3 containing ginsenoside Rf, to preventing itching of the scalp from showing more Excellent effect.
The effect assessment of [test example 10] increase potassium ion channel activity
Minoxidil as alopeciaing therapeutic agent is known as potential mitochondria K ~+Channel Opener (KATP 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 to prevent K using in the fibroblast for forming scalp coriumATPChannel Orinase (SIGMA AIDRICH, T0891) handled, to inhibit cell Proliferation, and be again turned on potassium ion Channel and restore cell Proliferation.
In order to evaluate the K as this compositionATPThe function of channel opener, present invention uses as fibroblast Mouse embryonic fibroblasts system (Mouse embryonic fibroblast cell line, NIH3T3) cell line of system. This cell line is with 3T3 scheme, to the fibroblast separated from NIH Swiss mouse embryo (Swiss mouse embryo) System carries out cell line obtained from nature immortalization.By the cell line in DMEM (Gibco BRL, Gai Se containing 10%FBS Regensburg, MD, the U.S.) in, maintaining 5%CO2, cultivate 24 hours in 37 DEG C of incubator.NIH3T3 is inoculated in 96 orifice plates, And after being cultivated 24 hours in 37 DEG C of incubator, handled with the orinase of 2.5mM, and after 10 minutes, point It is not used as at 10 μM of minoxidil of positive controls and the ginsenoside Rf of 2.5ppm, 5ppm and 10ppm concentration Reason, and by 48 hours after drug-treated, then ability of cell proliferation is measured using WST-1 kit (Roche (Roche)). And it the results are shown in following table 15.
Table 15
Classification Ability of cell proliferation (%)
Non-treated control group (control) 100
Minoxidil 132
Ginsenoside Rf (2.5ppm) 115
Ginsenoside Rf (5ppm) 121
Ginsenoside Rf (10ppm) 132
From above-mentioned table 15 it is found that when being handled with ginsenoside Rf, fibroblastic proliferation is restored, and cell Proliferative capacity depends on the concentration of the ginsenoside Rf of processing and increases, and can be confirmed at the ginsenoside Rf with 10ppm When reason, cell Proliferation is restored to level when being handled with minoxidil.
The melanin production facilitation effect of [test example 11] ginsenoside Rf is tested
The benzyl penicillin and 0.2 μM of terephthalic acid (TPA) of the fetal bovine serum, 100IU of addition 5% in RPMI culture medium (TPA) in culture medium, melanocyte is inoculated in 24 orifice plates with 50,000 cells/wells.Second day, to the thin of inoculation Born of the same parents use the ginsenoside Rf as substances of the ultimate density of 10ppm or 50ppm to handle.It will be with 0.1% DMSO The group of processing is used as negative control group as group, using the group for using 100 μM of IBMX to handle as positive controls, by above-mentioned each group It is cultivated three days at a temperature of 37 DEG C.After culture, with phosphate buffer (PBS) cleaning orifice, and it is added 100 μ l's in every hole After 1N NaOH, the melanin in cell is dissolved.Using plate culture assay instrument (microplate reader), at 405nm The absorbance (synergistic effect 2, microplate reader (VT, the U.S.)) of the dissolved melanin of measurement.It will promote the black of ginsenoside Rf The result that the effect of the generation of element is compared with control group is shown in following table 16.
Table 16
Sample B16 cell amount (%)
DMSO (0.1%) 100
IBMX(100μM) 120
Ginsenoside Rf (10ppm) 112
Ginsenoside Rf (50ppm) 123
It is found that ginsenoside Rf promotes the B16 cell of melanocyte, to increase melanin from above-mentioned table 16 It generates, therefore can show that the effect of excellent promotion melanin production.
The effect that [test example 12] ginsenoside Rf promotes transcription factor (MITF) and tyrosinase to express in melanocyte Fruit
It using 501mel cell line, is inoculated in 6 orifice plates with 500,000 cells/wells, and in each hole, with 0.1% The conduct positive controls of dimethyl sulfoxide (DMSO) processing, use that 100 μM of IBMX handles as positive controls, Yi Jiyong The conduct experimental group of 10ppm life saponin(e Rf processing, and after being cultivated 24 hours, 48 hours and 72 hours at a temperature of 37 DEG C, it obtains To protein.For the protein so obtained, western blot (Western is carried out using MITF and tyrosinase antibody 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 as 100, and be compared with the value and be shown in following table 17 In.
Table 17
It has been confirmed that ginsenoside Rf improves MITF and tyrosinase protein matter in melanocyte from above-mentioned table 17 Expression.
The antimicrbial power of [test example 13] ginsenoside Rf is evaluated
In order to evaluate the antimicrbial power of ginsenoside Rf, implement antibacterial experiment.Specific test method is as follows.
Staphylococcus aureus used in experiment (Staphylococcus aureus), Escherichia coli (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). The dilution that culture solution is diluted in each culture medium with 1/100 (albicans strain is using 1/10) is as test organisms Liquid come using.For aspergillus niger, 2 × 10 will be prepared into8The spore suspension of cfu/ml is used as test bacteria liquid.
The test bacteria liquid of 0.15ml and the mixed liquor that mixes are added in each culture medium of 15ml as dilute solution It uses.
In 96 orifice plate 1 row, 10ppm ginsenoside Rf is separately added into the amount of every 16 μ l of hole, and with every 184 μ l of hole Dilute solution is added in amount.Dilute solution is added with the amount of every 100 μ l of hole in other holes.The mixed liquor that No. 1 is arranged mixes equal After even, 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 3 Number row in.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.
After 48 hours, whether the increment of bacterium is confirmed by suspensibility and microscope, to determine minimum inhibitory concentration (MIC) value, and the results are shown in following table 18.
Table 18
As shown in above-mentioned table 18, it can be confirmed that ginsenoside Rf shows antimicrbial power to a variety of bacterial strains, and pass through this It can predict that ginsenoside Rf can work in composition as natural antiseptic agent or antibacterial agent a bit.

Claims (1)

1. ginsenoside RfThe purposes in the cosmetic composition for improving acne is being prepared as sole active ingredient.
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