CN107595659A - The application of Dermatologic preparation composition containing ginsenoside RG3 - Google Patents

The application of Dermatologic preparation composition containing ginsenoside RG3 Download PDF

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CN107595659A
CN107595659A CN201710970663.XA CN201710970663A CN107595659A CN 107595659 A CN107595659 A CN 107595659A CN 201710970663 A CN201710970663 A CN 201710970663A CN 107595659 A CN107595659 A CN 107595659A
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effect
composition
skin
ginseng
formulation example
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金东泫
柳权烈
李沃澯
廉明勋
曺濬喆
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Amorepacific Corp
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    • 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
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
    • A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
    • 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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/08—Solutions
    • 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
    • A61P17/08—Antiseborrheics
    • 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
    • A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • 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
    • A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the 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
    • A61Q7/00—Preparations for affecting hair growth

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Abstract

The present invention relates to the application of the Dermatologic preparation composition containing ginseng sapoglycoside Rg 3, said composition is using ginseng sapoglycoside Rg 3 as active ingredient, and improvement acne and allergic effect, dermostenosis and pore contractive effect can be provided, and the effect for improving skin complexion, promotion natural on-off cycles of hair growth can also be provided, improve white hair, antidandruff and corrosion-resistant effect.

Description

The application of Dermatologic preparation composition containing ginsenoside RG3
Related application
The application be Application No. 201480032940.7, the applying date be on April 24th, 2014, priority date be 2013 April 24, entitled " Dermatologic preparation composition containing ginsenoside RG3 " Chinese invention patent application division Application.
Technical field
The present invention relates to a kind of application of the composition containing ginseng sapoglycoside Rg 3, said composition can provide improvement acne With allergic effect, dermostenosis and pore contractive effect, and it can also provide and improve skin complexion effect, promote hair Growth, improve white hair, antidandruff and antiseptic effect.
Background technology
The skin of the mankind once defends film as human body and can play the change from temperature and humidity, ultraviolet, public hazards In the stimulation of the external environment conditions such as material protect intracorporeal organ function, skin with advancing age by internal factor and it is external because Element produces change.That is, internal factor is used for the secretory volume for the various secretion for adjusting metabolism, reduction immunocyte for reduction Function and cytoactive, so as to cause to make the biosynthesis of immune protein and raw body structural protein needed for live body to reduce. External factor is to cause to increase the ultraviolet content that earth's surface is reached in sunray by the destruction of ozone layer, and with environmental pollution Further in-depth and increase free radical and the harmful oxygen of activity etc. so that skin produces a variety of changes, i.e. skin thickness becomes Thin, increase wrinkle, and do not only result in and reduce elastic force and skin color is also dimmed, it is allergic often to occur, and increase Spot, freckle and senile plaque expelling, and make it that complexion variation, skin complexion are dimmed.
In order to prevent as in this skin and external factor caused by skin condition produce change to maintain healthy skin State, in the past in order to improve skin condition be made that following effort, i.e. by will from known various animals, plant, The physiological activator obtained in microorganism etc. is added to cosmetics and used.
Ginsenoside (Ginsenoside) is referred to as saponin(e possessed by ginseng, and wherein saponin(e is referred to as with sugar in chemistry One kind of the compound of body (glycoside).Ginsenoside be in order to the saponin(e with other plant makes a distinction and with from ginseng (ginseng) glycocide (glyco-side) of separation is the title that implication is enclosed.Ginsenoside can generally be named as ginseng Saponin(e Rx (ginsenoside-Rx) etc..Here, " R " is meant that root (Radix or Root), " x " is according to thin-layer chromatography (TLC) on the displacement (Rf values) of shown spot (spot) be named as successively from downside along upside o, a, b, c, d, e, f, G and h etc..Ginsenoside Endocrine system, immune system, metabolic system etc. headed by central nervous system bring influence, from And body function is adjusted and plays multi-effect.Up to now from the korean ginseng (including red ginseng) separation identify more than 30 kinds with On ginsenoside, and opened the pharmacological action of each ginsenoside successively.This ginsenoside shows similar work With or opposite effect.For example, the ginsenoside as typical saponin(e PPD, G-Rb1 Central nervous systems show to suppress effect Fruit, the G-Rg1 Central nervous systems as typical saponin(e PPT have facilitation effect.
Ginsenoside can be divided into following three kinds according to the structure of saponin(e aglycone (aglycon), i.e. diol type ginseng Saponin(e (Panaxadiol-type Ginsenosides：Ra1, Ra2, Ra3, Rb1, Rb2, Rb3, Rc, Rd, Rg3), three alcohol type people Join saponin(e (Panaxatriol-type Ginsenosides：Re, Rf, Rgl, Rg2), oleanolic acid type ginsenoside (Oleanolic acid-type Ginsenosides) etc..Diol type saponin content in ginseng is most, and it is 48%- 60%, secondly three alcohol type saponin contents account for 30%-35%, and contain micro oleanolic acid type saponin in ginseng.
As one kind in a small amount of ginsenoside for carrying out generating during steaming ginseng to this ginseng, the pharmacology about Rg3 Effect is with the related patent of following pharmacological action for oral use, i.e. has the effect of suppression cancer metastasis, platelet aggregation to suppress Effect and anti thrombotic action, vasorelaxation action, Experimental Hepatic Damage inhibitory action, the anti-medicine inhibitory action of anticancer etc..But It is that there is presently no the report for life saponin(e Rg3 is had the effect that as the composition of active ingredient, the i.e. combination Thing can improve acne and allergic effect, dermostenosis and pore contractive effect, improve skin complexion effect, promote hair Growth, improve white hair, antidandruff and antiseptic effect.
The content of the invention
Technical task
On the other hand, the inventors discovered that life saponin(e Rg3, which can be provided, improves acne and allergic effect, dermostenosis And pore contractive effect, there is provided improve skin complexion effect, promote natural on-off cycles of hair growth, improve white hair, antidandruff and antiseptic effect, and Complete the present invention.
Thus, it is an object of the invention to provide a kind of Dermatologic preparation composition containing ginseng sapoglycoside Rg 3, the skin Preparation composition for external use, which can provide, improves acne and allergic effect, dermostenosis and pore contractive effect, and can also Show to improve skin complexion effect, promote natural on-off cycles of hair growth, improve white hair, antidandruff and antiseptic effect.
Problem solves method
In order to achieve the above object, the present invention provides a kind of external preparation for skin for containing ginseng sapoglycoside Rg 3 as active ingredient Agent composition, said composition are the composition for improving acne.
In addition, the present invention provides a kind of Dermatologic preparation composition for containing ginseng sapoglycoside Rg 3 as active ingredient, the group Compound is the composition for improving skin complexion and the colour of skin.
In addition, the present invention provides a kind of Dermatologic preparation composition for containing ginseng sapoglycoside Rg 3 as active ingredient, the group Compound is the composition for pore refining.
In addition, the present invention provides a kind of Dermatologic preparation composition containing ginseng sapoglycoside Rg 3, said composition is for promoting Enter the composition of natural on-off cycles of hair growth.
In addition, the present invention provides a kind of Dermatologic preparation composition containing ginseng sapoglycoside Rg 3, said composition is for preventing The only composition of white hair.
In addition, the present invention provides a kind of Dermatologic preparation composition containing ginseng sapoglycoside Rg 3, said composition is antidandruff Composition.
In addition, the present invention provides a kind of Dermatologic preparation composition containing ginseng sapoglycoside Rg 3, said composition is used as natural Preservative.
Invention effect
The composition of the present invention contains ginseng sapoglycoside Rg 3 and can provide improves acne and allergic effect, skin receipts Tight and pore contractive effect, and improvement skin complexion effect can be provided, promote natural on-off cycles of hair growth, improve white hair, antidandruff and prevent Rotten effect.
Embodiment
Contain the ginseng sapoglycoside Rg 3 (Ginsenoside Rg3) as active ingredient according to the composition of the present invention.
The ginseng sapoglycoside Rg 3 that the present invention uses has the structure of formula 1 below.
Chemical formula 1
The present invention ginseng sapoglycoside Rg 3 can be extracted from plant, can also use according to method well known in the art come Synthesis, or ginseng sapoglycoside Rg 3 on the market.In addition, ginseng sapoglycoside Rg 3 can obtain from ginseng extract. Now, ginseng species used in being not particularly limited, water ginseng, red ginseng, white ginseng, Tai Ji ginseng, ginseng palpus can be used Deng.In addition, the ginseng extract not only includes the leachate for being leached from ginseng, transferring and obtaining, but also including right again The leachate concentrate that either whole leachate is concentrated and obtained in part can also be included to the concentrate again Being contained in for being dried and manufacture is stagnant, decoction, tincture, in fluid elite and ginseng to play the chemicals of main efficacy results Matter, it is of course also possible to which including plant in itself, the extraction of all sites of ginseng such as stem, root, leaf, flower, fruit can be used Thing, and it is not limited to the extract of some privileged site.In addition, for extracting ginseng sapoglycoside Rg 3 from ginseng extract Method can use known method.
Specifically, after preparing ginseng extract using water or organic solvent from ginseng by means commonly known in the art, The ginseng sapoglycoside Rg 3 is separated from the ginseng extract.The organic solvent that the present invention uses can from ethanol, methanol, butanol, Select in the group of ether, ethyl acetate, chloroform and the mixed solution of these organic solvents and water composition, and preferably use 80% ethanol.Now, Extracting temperature is preferably 10-80 DEG C, can extract 3-24 hours.When the disengaging Extracting temperature and carry When taking time range, extraction efficiency may be reduced or produce the change of composition.
Preferably, the content of the ginseng sapoglycoside Rg 3 contained in composition of the invention is in the total content of composition Account for 0.001-50 weight %.This is due to when the content of the active ingredient is not up to 0.001 weight %, by described into separating The effect of rising and low effort, cause to produce cutaneous safety or formulation when more than 50 weight %The problem of upper.
The composition of the present invention may be used as improving the Dermatologic preparation composition of acne, skin preparations for extenal use combination Thing can provide antibacterial effect, particularly excellent to the antibacterial effect of acne pathogenic bacteria, and provide antiphlogistic effects.
The composition of the present invention may be used as improving the Dermatologic preparation composition of complexion and the colour of skin, by the skin When preparation composition for external use is used on skin, by expanding capillary and stimulating circulation, so as to by nutritional ingredient Smoothly feed to skin, suppress skin aging and there is superior complexion and colour of skin improvement.
The present invention composition may be used as can pore refining, adjust sebum and improve allergic skin preparations for extenal use Composition, when the Dermatologic preparation composition is used on skin, the sebum of excessive secretion can be suppressed, by promoting to live Property the removal of oxygen and the synthesis of collagen and can pore refining, and by reducing the performance of inflammatory factor and with superior The allergic effect of suppression.
The composition of the present invention may be used as promoting the composition of natural on-off cycles of hair growth, and said composition is stopped by promoting Only the hair cycles of phase are transitioned into the hair cycles in growth period, can promote what natural on-off cycles of hair growth and kainogenesis were grown so as to not only have Effect, but also with the effect that can grow existing hair healthy, and prevention is provided and suppresses hair and is taken off from scalp The phenomenon or hair sparse that fall, attenuate phenomenon.
The composition of the present invention may be used as preventing the composition of white hair, and said composition is by increasing melanocyte MIFT expression and melanocyte activity and to promote the synthesis of melanin, so as to prevent the generation of white hair, And promotion is provided and induces effect caused by dark hair.
The composition of the present invention may be used as the Dermatologic preparation composition of antidandruff, and said composition can be effectively discharged out Accumulate and purify scalp in the toxin on hair and scalp, and suppress dandruff bacillus propagation and growth and prevent head Skin inflammatory reaction, the anti-oxidation efficacy of generation and effect in addition with superior inhibitory activity oxygen and calm and strengthen scalp, So as to provide the effect for strengthening original defence capability.
The present invention composition may be used as natural antiseptic agent composition, said composition due to containing natural component, so as to Superior antiseptic effect and harmless effect are provided.
The medium being licensed on cosmeceutical or Dermatology or matrix can be contained according to the composition of the present invention Into formulation.Said composition is as all formulations being suitable for locally, such as can be provided as solution, gel, solid, paste The anhydrous product of shapeIn the liquid phase disperse oil phase and obtain emulsion, suspension, microemulsion, Microcapsules, microparticle ball the form of the vesica dispersant of ionic (liposome) and nonionic or can also either provide For the form of face cream, toner, skin care milk, foundation cream, ointment, spraying or concealer.Further, it is also possible to use foam (foam) shape Or the form of the aerosol composition of the propellant further containing compression.These compositions can be by usual in this area Method and manufacture.
Particularly, Dermatologic preparation composition of the invention is used for anti-antidandruff, educates hairOr prevent white hair When, its formulation can be made for the form of scalp and the composition of hair, and be not particularly limited formulation, for example, can be with Formulation turn to preparation for baldness, hair nutrition toner, scalp nursing solution, hair nursing liquid, shampoo, hair conditioner, hair breast or Scalp and hair dual-purpose conditioning liquid etc..
In addition, fatty material, organic solvent, lytic agent, concentrating agents, gelation can be contained according to the composition of the present invention Agent, softening agent, antioxidant, suspending agent, stabilizer, foaming agent (foaming agent), aromatic, surfactant, water, Ionic or nonionic softening agents, filler, metal ion block agentChelating agentAntistaling agent, vitamin, blocking agent, wetting agent, essential oil, dyestuff, pigment, hydrophily or lipophile are lived Property agent, lipid vesicle or any other compositions with being generally used in cosmetics it is identical and led in cosmeceutical or Dermatology The usually used adjuvant in domain.The amount of the adjuvant added is what is generally used in cosmeceutical or Dermatology field Amount.
In addition, the composition of the present invention can contain the material for promoting skin to absorb to increase skin improvement effects.
Embodiment
Hereinafter, the structure and effect of the present invention are further illustrated by experimental example and formulation example.But these Test example and formulation example are only the example for being used to help understand the present invention and offer, and scope of the invention and scope are not limited to Following examples.
[reference example 1] prepares personal saponin(e Rg3
In order to which the personal saponin(e Rg3 used the effect of testing the composition of the present invention is from AMBO research institutesPurchase.
[formulation example 1 and compare formulation example 1]
Nutrition face cream (unit has been manufactured by usual way according to the composition of table 1 below：Weight %).
Table 1
[table 1]
[test example 1] improves complexion effect
In order to evaluate the promotion skin blood circulating effect of the cosmetic composition according to the present invention, LDPI (Laser are utilized Doppler Perfusion Imager；Periscan PIM II, Perimed (stochholm, Sweden)) determine skin In blood circulation degree.What LDPI was widely known by the people is as the sanguimotor equipment in measure skin and to be used at present Equipment, it is a kind of blood flow velocity that can not only be determined in capillary of skin and amount but also can determine parteriole With the very sensitive equipment of the flowing in veinlet.
Adapted to 30 minutes after utilizing soap washed in constant temperature and humidity room, and initial value is determined using LDPI.First, The forehead of 30 cool women of usually trick is determined with the initial CBF of lower part using LDPI.Subject is made afterwards With the formulation example 1 of one week and compare formulation example 1, so that by the CBF determined compared with the initial measured value And the result (skin blood flow change) drawn is shown in table 2 below.
Table 2
[table 2]
It can be derived that from the result of the table 2, according to the cosmetic composition of the present invention compared to not containing ginseng soap Glycosides Rg3 comparison formulation example 1 significantly increases skin blood flow, and is able to confirm that by this sanguimotor promotion Improve complexion.This finally shows effectively transmit skin according to the cosmetic composition containing ginseng sapoglycoside Rg 3 of the present invention The nutritional ingredient of skin, and suppression and the aging of interest for delinquency skin.
[test example 2] improves colour of skin effect
For the improvement colour of skin effect learnt the formulation example 1 He compare formulation example 1, the subjects of 30 are made respectively With after (evening 1 times/day coating, 1 week altogether), the colour of skin using Facial Stage DM-3 (Moritex, Japan) appraisal of equipment Improvement degree.Colour of skin improvement rate judges the lightness changing value of skin and face using the lightness measured value and color measurement of skin Color change value, and its result is shown in table 3 below.Lightness changing value and the big expression of color change value improve the colour of skin.
Table 3
[table 3]
It can determine from the result of the table 3, according to the comparison formulation example 1 for not containing ginseng sapoglycoside Rg 3 of the present invention The effect of not showing to improve the colour of skin, reverse side, the skin in the formulation example 1 of the ginseng sapoglycoside Rg 3 containing active ingredient after use Color ratio improves a lot before using.
[test example 3] pore refining effect
1. the effect of the pore refining by promoting collagen biosynthesis
By comparing ginseng sapoglycoside Rg 3 and TGF-β according to the present invention, promote collagen biosynthesis so as to determine Effect.First, (seeding) 10 is inoculated with each hole of 24 orifice plates (well)5Individual fibroblast (fibroblast), and It is trained the degree of cells grown to 90%.After plasma-free DMEM medium culture 24 hours, serum-free training will be dissolved in Support the ginseng sapoglycoside Rg 3 of the invention in base and TGF-β is processed into 10ng/ml respectively, and in CO2It is small that 24 have been cultivated in culture medium When.The upper liquid of above-mentioned solution is taken out, and using I procollagen types ELISA kit (procollagen type (I)；# MK101, TAKARA (Shiga, Japan)) observation precollagen (procollagen) increase and decrease whether.Its result is shown in table 4 Go out, the synthesis capability of collagen is with being set as that 100 untreated fish group is contrasted.
Table 4
[table 4]
Substances Collagen synthesis ability (%)
Untreated fish group 100
TGF-β 183.5
Ginseng sapoglycoside Rg 3 197.2
It can determine from the result of the table 4, according to TGF-β of the ginseng sapoglycoside Rg 3 of the present invention than positive controls Show higher levels of excellent collagen synthesis ability.Thereby, it is possible to determine the ginseng sapoglycoside Rg 3 energy according to the present invention It is enough to shrink the pore for becoming big by increasing the collagen growing amount around pore.
2. pore refining effect
For the pore refining effect learnt formulation example 1 He compare formulation example 1, following evaluation has been carried out.Select pore big Testee men and women 20, be divided into two groups and every group of 10 people, each group is coating the formulation example 1 of totally 4 weeks on the face respectively daily With the nourishing cream for comparing formulation example 1.Pore refining effect is judged by being taken a picture respectively and by expert before testing and after 4 weeks With the naked eye evaluate and realize.Its result shows (opinion rating in table 5 below：0- does not shrink completely；5- shrinks a lot).
Table 5
[table 5]
Substances Opinion rating
Formulation example 1 3
Compare formulation example 1 0
It can determine from the result of the table 5, compare ungauged regions pore effect in formulation example 1, but in formulation example 1 Situation is shown to the naked eye determine the pore refining effect of degree, excellent so as to be had according to the ginseng sapoglycoside Rg 3 of the present invention The effect of good reduction pore size.
[test example 4] suppresses sebum secretion effect
1. suppress the effect of skin excessive secretion by suppressing 5α-reductase activity
In order to determine to suppress the effect of 5α-reductase activity, determine in HEK293-5 α R2 cells [14C] testosterone is converted into [14C] protona (DHT:Dihydrotestosterone ratio).P3 × FLAG-CMV-5 α are transfected in HEK293 cells R2, and add it in 24 orifice plates and add 2.5 × 10 in each hole5Individual cell (Park et al., 2003, JDS.Vol.31, pp.191-98) and cultivated.Second day, it is replaced with the new culture added with enzyme matrix and inhibitor Base.The matrix of culture medium employ 0.05 μ Ci [14C] testosterone (Amersham Pharmacia biotech, UK).
In order to determine to suppress the degree of 5α-reductase activity, ginseng sapoglycoside Rg 3 is added and in 37 DEG C, 5% CO2Training Support and cultivated 2 hours in base.Now, the nutrient solution for not adding ginseng sapoglycoside Rg 3 is used as negative control group, by addition have it is non-that The nutrient solution of male amine (finasteride) is used as positive controls.Afterwards, nutrient solution is extracted, and utilizes 800 μ l ethyl acetate Steroids is extracted, the organic solvent layer on top is separated afterwards and it is dried, use 50 μ l ethyl acetate again afterwards Remaining residue is dissolved, by it in silastomer plate silica gel 60F254 (Silica plastic sheet kieselgel With ethyl acetate-nucleic acid (1 on 60F254):1) deployed as solvent.
Dry plastics sample in atmosphere, Bath system is employed afterwards for determining the content of isotope, wherein, will Dry plastic sample and X-ray film are together added in Bath box, and the testosterone resided on film was determined after one week With the isotopic content of protona, conversion ratio and inhibiting rate are calculated respectively according to following mathematical expressions 1 and mathematical expression 2 afterwards, Its result is shown in table 6 below.
Mathematical expression 1
Mathematical expression 2
Table 6
[table 6]
Substances Conversion ratio (%) Inhibiting rate (%)
Negative control group 48 -
Positive controls 27.6 42.5
Ginseng sapoglycoside Rg 3 16.3 66.0
Can determine from the result of the table 6,5α-reductase by testosterone be converted into protona and with it is intracellular Accommodate body protein to combine and enter in core so that sebocyte cell activates and promotes to break up, so that 5α-reductase plays The effect of excessive secretion sebum in sebaceous glands, and can effectively suppress the activity of 5α-reductase by ginseng sapoglycoside Rg 3, from And block testosterone to be converted into protona, and show with than can suppress 5α-reductase activity known to it is non-that The more excellent inhibition of male amine.Thus, ginseng sapoglycoside Rg 3 can suppress skin by effectively suppressing the activity of 5α-reductase The excessive secretion of fat.
2. suppress sebum secretion effect
In order to draw above-mentioned formulation example 1 and compare the effect for suppressing sebum secretion of formulation example 1, following evaluation has been carried out. The subject men and women 30 more than sebum secretion is thought in selection, and is coated on the position specified daily in facial skin 4 weeks Formulation example 1 and the nutrition face cream for comparing formulation example 1.The judgement of sebum minimizing effect is by using sebum flowmeter (Sebumeter SM810, C+K Electronic Co., Germany) determines being averaged after two weeks and after four weeks respectively Sebum slip (%) and realize, its result is shown in table 7 below.
Table 7
[table 7]
It can determine from the result of the table 7, according to the ginseng sapoglycoside Rg 3 contained as active ingredient of the present invention Formulation example 1 more effectively suppresses the excessive secretion of sebum than not containing the comparison formulation example 1 of the ginseng sapoglycoside Rg 3.
[formulation example 2 and compare formulation example 2-3]
Composition and content (weight %) according to being shown in table 8 below are prepared for formulation example 2 and compare formulation example 2-3.Tool Body is described as follows, i.e. formulation example 2 coordinates ginseng sapoglycoside Rg 3, compares formulation example 2 and is entirely free of for improving acne-prone skin Active ingredient, compare formulation example 3 and contain as the standard substance for the benchmark for judging antibacterial ability more as acne therapeutic agent Erythromycin (erythromycin).
Formulation example 2 and the manufacture method for comparing formulation example 2-3 are as follows.The composition being completely dissolved in the A of table 8 below, and The composition being completely dissolved in other dissolving tank in B, afterwards, B is added in A and carries out mixed dissolution.Here, by C into Point according to the mixing ratio described in table 8 be added and it is well mixed after filter, so as to produce this composition.
Table 8
[table 8]
[test example 5] is tested the antibacterial ability of acne bacterium
Using by the formulation example 2 and formulation example 2-3 composition is compared each cosmetic composition for preparing, to making For the propionibacterium acnes (ATCC 6919 of acne pathogenic bacteria：Culture medium-BHI fluid nutrient mediums (broth)) carry out antibacterial energy Power is tested.
It is as follows to the antibacterial ability test method of acne bacterium.
(1) experiment bacterium solution is prepared
Employ by the way that propionibacterium acnes is inoculated with to carry out the culture medium of Anaerobic culturel in BHI fluid nutrient mediums.
(2) dilute solution is prepared
Addition 0.15ml's is described in 15ml BHI fluid nutrient mediums (pH6.8) or LB fluid nutrient mediums (pH4.5) Test bacterium solution and be sufficiently mixed, and its solution is used as dilute solution.
(3) sample is prepared
By formulation example 2 sample will be directly used as with the cosmetic composition stoste for comparing formulation example 2-3 preparations.
(4) antibacterial ability is tested
1) added in the Tissue Culture Plate (96well plate) in 96 holes and meet the sample of initial concentration and be separately added into Dilute solution to total amount is 200 μ l.
2) it is sufficiently mixed after the mixed liquor of the first row, 100 μ l of extraction mixed liquor is added to the second row and fully mix Close, extract the form execution doubling dilution (double dilution) that 100 μ l are added to the third line again afterwards.
3) quiescent culture after 24 hours and 48 hours at 32 DEG C, judge whether bacterium is proliferated into suspended degree, and by bacterium Cmin without propagation is defined as MIC (minimal inhibitory concentration, Minimum Inhibitory Concentration) value.Such as Fruit mixed liquor is muddy and is difficult to judge when whether bacterium breeds, it is necessary to be confirmed by micro- sem observation.
Shown for the antibacterial ability result of the test of acne bacterium in table 9 below.MIC, which is converted into, is contained in having in formulation Imitate the concentration of composition and be marked.
Table 9
[table 9]
Project pH Propionibacterium acnes
Formulation example 2 5.7 >48ppm
Compare formulation example 2 5.7 Maximum concentration (no antibacterial ability)
Compare formulation example 3 5.7 >100ppm
It can determine from the result of the table 9, ppm concentration is smaller in MIC, can represent the antibacterial energy to acne bacterium The more effective material of power, in situation and the comparison formulation example 3 using the erythromycin for being known as acne therapeutic agent of formulation example 2 Compare, ppm concentration is substantially reduced, and therefore, the composition containing ginseng sapoglycoside Rg 3 has more excellent antibacterial energy to test organisms Power.
[test example 6] suppresses the experiment of lipid synthesis (Lipogenesis)
Using as the 3T3-L1 cells of the fibroblast (fibroblast cell line) of mouse living with 1 × 105Carefully The amount in born of the same parents/hole is attached on 6 well culture plates (culture plate), in 6 well culture plate be equipped with containing hyclone (FBS, Fetal bovine serum) DMEM (Dulbecco's modified eagle's medium, GIBCO BRL, life section Skill company) culture medium.DMEM (containing the 10% FBS) culture mediums that more renewed after 2 days and cultivate 2 days again.Afterwards, lead to Cross the insulin (insulin) containing 1 μ g/mL, 0.5mM IBMX and 0.25 μM of dexamethasone (dexamethasone) DMEM (contains 10% FBS), induces the cell of above-mentioned culture to be broken up again, and using 50 μM ginseng sapoglycoside Rg 3 and Caffeine processing, after 2 days, is replaced with containing the DMEM of insulin and cultivates 5 days again.After 5 days, it is replaced with again Normal incubation medium (DMEM, contains 10% FBS), and observation culture to above-mentioned cell becomes lipoblast form.
Suppress to accumulate the effect of fat in adipocyte to evaluate ginseng sapoglycoside Rg 3, utilize above-mentioned differentiated 3T3- L1 adipocytes have carried out soudan III dyeing (S4136, sigma-aldrich).At normal temperatures by adipocyte with 4% it is more Polyformaldehyde (pH 7.2) is fixed in phosphate buffer, afterwards with PBS (phosphate buffered saline) to it Washed, taken a picture after being dyed afterwards with soudan III, so as to by visually comparing.Control group, which only used, to be not added with Substances or the culture medium for comparing material, the caffeine for using 50 μM as other comparative groups are handled.Suppress lipopexia Degree is by the way that dye levels are divided into +++, ++ ,+,-and divided rank.Now, more tend to +++, it represents that dye levels are bigger. Its result is shown in table 10 below.
Table 10
[table 10]
Sample Inhibiting rate %
Control group +++
Comparative group +
Ginseng sapoglycoside Rg 3 -
It can be learnt from the result of upper table 10, the ginseng sapoglycoside Rg 3 used in the present invention not only makes in adipocyte The fat content of accumulation is few, and has more excellent suppression compared to the caffeine for the material for being known as suppressing lipid synthesis The effect of lipid synthesis processed.Thus, sebum is reduced by suppressing the synthesis of lipid, so as to suppress the generation of acne.
[test example 7] improves acne, reduces sebum secretion and have non-stimulated experiment
Carbuncled 30 experimenters are divided into 3 groups with every group 10, corresponding testee in each group is made respectively With 1 month by above-mentioned formulation example 2, compare the cosmetic composition of formulation example 2-3 preparation.The yardstick for improving acne is set as 1 Divide to 5 points, wherein, 1 point represents " not having ", and 3 points represent " general ", and 5 points represent " very good ".Its result of the test is in table 11 below In marked with the average mark of 10.
Acne date of the period to judge to disappear that disappear is used as benchmark, and acne, which recur, uses " whetheing there is " and with after 1 month As a result it is used as benchmark.The reduction of sebum secretion is set as 1 point to 5 points, wherein, 1 point represents " not having ", and 3 points represent " general ", 5 Divide and represent " very good ".Its result of the test is marked in table 11 below with the average mark of 10.Skin irritatin is whether there is by (having The number of stimulate the reaction)/(overall test person number) expression.
Table 11
[table 11]
It can be learnt from the table 11, the formulation example 2 compared with of formulation example 2 does not recur again compared to acne, right on the whole Improving acne has excellent effect.In addition, the situation in the comparison formulation example 3 containing antibacterial ability standard substance shows and changed The effect of kind acne, but it is big to skin irritatin during use, therefore be not suitable for long-term use, and according to the combination of the present invention Thing shows due to non-stimulated and also is adapted for long-term use.
[formulation example 3 and compare formulation example 4]
Shampoo has been manufactured using the composition of table 12 below.Specifically, surfactant and second two are added in purified water Alcohol distearate, and 80 DEG C are heated to after equably dissolving, 40 DEG C are slowly cooled to by stirring, in said mixture Middle input is mixed according to the active ingredient and preservative, viscosity modifier, spices and hair conditioner of the present invention, by stirring Mix and be cooled to room temperature, prepared so as to complete.
Table 12
[table 12]
Composition (weight %) Formulation example 3 Compare formulation example 4
Ammonium dodecyl ether sulfate 10 10
Polyoxyethylene lauryl ether ammonium sulfate 5 5
Cocoamidopropyl betaine 2 2
EGDS 1.5 1.5
Cocoyl MEA 0.8 0.8
Ginseng sapoglycoside Rg 3 5.0 -
Polyquaternium-10 0.2 0.2
Blueness 1 0.0002 0.0002
Yellow 4 0.0001 0.0001
Methyl p-hydroxybenzoate 0.1 0.1
Spices 0.8 0.8
Citric acid 0.1 0.1
Dimethyl silicone polymer 1.0 1.0
Water To 100 To 100
[test example 8] reduces dandruff effect test
The more male of 24 dandruffs of 19-35 year is selected, and is divided into two groups with every group 12, each group is passed through respectively Formulation example 3 and the shampoo that compares formulation example 4 of the in the following manner using 1 month, then determine dandruff slip.
After being had one's hair wash before on-test with common in general shampoo, dandruff of the collection by accumulation in 2 days, and by collection The weight of dandruff with by each formulation example 3 and compared with formulation example 4 shampoo to have one's hair wash once two days and after completing experiment The weight of dandruff by accumulation in 2 days is compared and evaluated.Now, the dandruff of accumulation is direct by vacuum suction apparatus Gathered from scalp, and dandruff slip is obtained according to following mathematical expression 3, its result is shown in table 13 below.
Mathematical expression 3
Table 13
[table 13]
It can be learnt from the result of the table 13, the situation of the formulation example 3 containing ginseng sapoglycoside Rg 3 shows excellent The effect of antidandruff.
[test example 9] prevents the effect test of pruritus of scalp
24 more serious men and women of scalp itch are felt in selection 25-45 year, are divided into two groups with every group 12, and use respectively After formulation example 3 once uses 2 weeks with the shampoo for comparing formulation example 4 and with every 3 days, by preventing that itching of the scalp effect from being to comment Valency benchmark and evaluated, its result is shown in table 14 below.
[metewand]
It is very outstanding -5 points
Outstanding -4 points
Typically -3 points
Bad -2 points
It is very bad -1 point
Table 14
[table 14]
Classification Formulation example 3 Compare formulation example 4
The pruritus removal effect of scalp 4.3 2.3
It can be learnt from the result of the table 14, the situation of the formulation example 3 containing ginseng sapoglycoside Rg 3 is to preventing scalp scabies Itch disease effect it is more excellent.
[test example 10] increases the effect assessment of potassium ion channel activity
Minoxidil as alopeciaing therapeutic agent is known potential mitochondria K ~+Channel Opener (KATP Channel opener), it is the representative drugs for treating male sex hormone alopecia.In order to evaluate the machine of this minoxidil System, employs tests below method：For being formed in the fibroblast of scalp corium, blocking K is usedATPThe toluene of passage Sulphur butyl urea (SIGMA ALDRICH, T0891) handles and suppresses cell propagation, is again turned on potassium-channel afterwards and recovers thin Born of the same parents breed.
In order to evaluate the conduct K of this compositionATPThe function of channel opener, it is employed herein as into fiber finer NIH3T3 (Mouse embryonic fibroblast cell line) cell line of born of the same parents.The cell line is by from NIH Switzerland In mice embryonic (Swiss mouse embryo) fibroblast of separation using 3T3 experimental methods ( Protocol the cell line of nature immortalization) is carried out.DMEM (the Gibco containing 10% FBS are added in the cell line BRL, Gaithersburg, MD, the U.S.), and in 5% CO2, maintain to have cultivated 24 hours in 37 DEG C of incubator.Will NIH3T3 is added in 96 orifice plates, and after having cultivated 24 hours in 37 DEG C of incubator, at 2.5mM orinase Reason, after 10 minutes will use respectively as positive controls 10 μM of minoxidil handle, concentration respectively be 2.5ppm, 5ppm and 10ppm ginseng sapoglycoside Rg 3 processing.After drug-treated 48 hours, determined using WST-1 kits (Roche) thin Born of the same parents' multiplication capacity.As a result shown in table 15 below.
Table 15
[table 15]
Classification Ability of cell proliferation (%)
Untreated control group (Control) 100
Minoxidil 132
Ginseng sapoglycoside Rg 3 (2.5ppm) 114
Ginseng sapoglycoside Rg 3 (5ppm) 123
Ginseng sapoglycoside Rg 3 (10ppm) 129
It can determine from the result of the table 15, recovered fibroblast in the situation for having handled ginseng sapoglycoside Rg 3 Propagation, and ability of cell proliferation dependent on processing ginseng sapoglycoside Rg 3 concentration and increase, at by ginseng sapoglycoside Rg 3 Cell breeds the level reverted to when handling minoxidil in the case of managing into 10ppm.
The effect test for promoting melanin generation of [test example 11] ginseng sapoglycoside Rg 3
The culture that the hyclone of addition 5%, 100IU benzyl penicillin and 0.2 μM of TPA are formed in RPMI culture mediums In base, by melanocyte (melan-a) in 24 hole microtiter plates (24-well microtiter plate) plant division into 50,000 cells/wells.Second day, the ginseng sapoglycoside Rg 3 processing as substances, the ginseng soap are used in the cell of plant division Glycosides Rg3 ultimate density is 10ppm or 50ppm, and 0.1% DMSO processing is used as negative control group, as positive right According to 100 μM of IBMX processing of group use, cultivated afterwards at a temperature of 37 DEG C 3 days.After culture, with PBS washing holes and difference Add 100 μ l 1N NaOH and dissolve intracellular melanin.Using ELIASA (microplate reader) in 405nm Determine the absorbance (Synergy2, BioTek (VT, USA)) of the melanin of dissolving.The promotion melanin generation of ginseng sapoglycoside Rg 3 The result that effect is drawn compared with control group is shown in table 16 below.
Table 16
[table 16]
Sample B16 cell amount (%)
DMSO (0.1%) 100
IBMX(100μM) 120
Ginseng sapoglycoside Rg 3 (10ppm) 110
Ginseng sapoglycoside Rg 3 (50ppm) 120
It can be learnt from the result of the table 16, ginseng sapoglycoside Rg 3 promotes the synthesis of melanin and increases melanin life Into so as to show the excellent effect for promoting melanin generation.
MITF and tyrosinase (tyrosinase) are expressed in the promotion melanocyte of [test example 12] ginseng sapoglycoside Rg 3 Effect
Using 501mel cell line in 6 hole microtiter plates (6-well microtiter plate) plant division into 500,000 cells/wells, in each hole, 0.1% DMSO processing is used as negative control group, is made as positive controls With 100 μM of IBMX processing, the ginseng sapoglycoside Rg 3 for using 10ppm as test group is handled, so as to be cultivated at a temperature of 37 DEG C Protein is obtained after 24 hours, 48 hours, 72 hours.Using MITF and tyrosinase antibody to thus obtained albumen Matter has carried out Western blotting.The extraction of protein and the protein imprinted standard method used by usual technical staff are real It is existing.Result after Western blotting compared with being set to 100 value of negative control group after shown in table 17 below.
Table 17
[table 17]
It can determine from the result of table 17, ginseng sapoglycoside Rg 3 can increase MITF and tyrosinase in melanocyte Protein expression.
The antibacterial ability evaluation of [test example 13] ginseng sapoglycoside Rg 3
Antibacterial tests have been carried out in order to evaluate the antibacterial ability of ginseng sapoglycoside Rg 3.Specific test method is as follows.
Staphylococcus aureus (Staphylococcus aureus), Escherichia coli used in experiment (Escherichia coli), Pseudomonas aeruginosa (Pseudomonas aeruginosa) bacterial strain are in pancreas protein soybeans culture medium Cultivated in (Tryptic Soy Broth), and Candida albicans (Candida albicans), aspergillus niger (Aspergillus niger) bacterial strain is using gauze in Sharpe fluid nutrient medium (Sabouraud's Dextrose Broth) Cultivated.Nutrient solution is diluted to 1/100 (Candida albicans is diluted to 1/10) in each culture medium, the dilution is used It is tested bacterium solution.Aspergillus niger is will to be manufactured into 2 × 108Cfu/ml spore suspension is used as experiment bacterium solution.
In 15ml each culture medium add 0.15ml experiment bacterium solution and well-mixed solution is used as dilute solution.
16 μ l 10ppm ginseng sapoglycoside Rg 3 and 184 is separately added into 96 orifice plates (96well plate) the first row μ l dilute solution.100 μ l dilute solution is added in remaining hole.After the mixed liquor of the first row is sufficiently mixed, 100 μ l are taken It is added in the second row and is sufficiently mixed, takes 100 μ l to be added in the third line afterwards, diluted respectively twice by the above method.
Staphylococcus aureus (Staphylococcus aureus), Escherichia coli (Escherichia coli), green pus Bacillus (Pseudomonas aeruginosa) is cultivated in 32 DEG C of thermostat, and Candida albicans (Candida Albicans), aspergillus niger (Aspergillus niger) is cultivated in 25 DEG C of thermostat.
After 48 hours, by suspension degree and microscope confirm bacterium propagation whether, and determine MIC (MIC), its result is shown in table 18 below.
Table 18
[table 18]
Can determine from the result of the table 18, ginseng sapoglycoside Rg 3 goes out antibacterial ability to a variety of strains expresseds, thus, it is possible to Enough predict ginseng sapoglycoside Rg 3 and may be used as natural antiseptic agent or antiseptic in composition.

Claims (2)

1. the Dermatologic preparation composition for containing ginseng sapoglycoside Rg 3 as active ingredient is preparing the cosmetic of improvement complexion and the colour of skin Application in product composition.
2. ginseng sapoglycoside Rg 3 answering in the cosmetic composition for improving complexion and the colour of skin is prepared as unique active ingredient With.
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