CN103796631A - Compositions for whitening skin comprising madecassoside - Google Patents
Compositions for whitening skin comprising madecassoside Download PDFInfo
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- CN103796631A CN103796631A CN201280021188.7A CN201280021188A CN103796631A CN 103796631 A CN103796631 A CN 103796631A CN 201280021188 A CN201280021188 A CN 201280021188A CN 103796631 A CN103796631 A CN 103796631A
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- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
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- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
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Abstract
The present invention relates to a skin whitening composition comprising madecassoside as an active ingredient. The composition comprising madecassoside can be safely on the skin without causing side effects and has excellent effects of inhibiting the UV-induced production of skin darkening cytokine in keratinocytes and inhibiting the production of melanin in melanocytes to inhibit pigmentation. Thus, the composition comprising madecassoside as an active ingredient is very effective in inhibiting UV-induced pigmentation, inhibiting dark skin discoloration and whitening the skin.
Description
Technical field
The present invention relates to the skin lightening compositions and the pharmaceutics compositions thereof that contain asiaticoside, relate in particular to the skin lightening compositions that contains asiaticoside, the product stability excellence of said composition, can use safely skin is without any side effects, and check melanin generates and prevents pigmentation effect excellence.
Background technology
People's the colour of skin depends on melanin (melanin) concentration and the distribution of skin inside, but affected by the environment such as solar ultraviolet or fatigue, pressure or physiological condition.Be phenols polymer substance by the melanin generating in human body skin melanocyte, this phenols polymer substance has the combined state of black pigment and protein, interdicts thereby bringing into play the important function that is caused skin injury by ultraviolet.It was reported, the effect of tryrosinase that is stored in melanocyte in melanic biosynthesis is the most important.Tryrosinase (tyrosinase) is by being converted to intermediate product-be DOPA (DOPA) that melanin polymer generates, DOPA quinone (dopaquinone)-and bring into play central role in skin melanism process as amino acid whose a kind of tyrosine (tyrosine).But, become knownly although prepare melanic generation approach, resolve not yet in detail clear about tryrosinase effect with the synthetic induction mechanism of the melanin of last stage.（Korean?J.Medicinal?Crop?Sci.18(2):79.85.2010）。
On the other hand, up to now the multiple whitening material of exploitation in melanin is synthetic as central role, restraint of tyrosinase and a series of melanin route of synthesis afterwards, thus bring into play whitening effect.But the research direction of developing recently whitening agent is, not only the melanin of check melanin cell itself generates, and adjacent with melanocyte, and suppress to cause that by ultraviolet secretion melanin generates the effect of the keratinocyte (Keratinocytes) that promotes the factor, and then performance whitening effect.
Interaction between keratinocyte and melanocyte is participated in by various cytokines.It was reported, especially the PGE2 between them, by the cAMP of secretion volume increase melanocyte in glial cell, is bringing into play and is promoting melanic Main Function.The material that can suppress this PGE2 also can be applicable to effectively defend the whitening agent of UV-induced cutaneous pigmentation.This melanin synthesizes in skin too much and produces, will cause skin color dimmed, produce black speck, freckle etc.Therefore, suppress melanin in skin synthetic, not only can realize making that the colour of skin brightens, skin whitening, can also improve by ultraviolet, hormone and gene and cause producing the cutaneous pigmentation such as black speck, freckle disease.
On the other hand, used in the past and as hydroquinone (hydroquinone) or ascorbic acid usp/bp (ascorbic acid), kojic acid (kojic acid), glutathion (glutathione) etc., sieve propylhomoserin enzyme is had to the active material of inhibition and improved skin-whitening or cutaneous pigmentation disease.But hydroquinone is brought into play specific whitening effect, but its skin irritation is strong, thereby has the problem that infiltration capacity must be restricted to minute quantity; Because ascorbic acid usp/bp is easily oxidized, cause the cosmetic material variable color of its preparation, the problem such as rotten; Kojic acid is dangerous in solution, therefore exists cosmetic material preparation technology to become complicated shortcoming.In addition, the sulfhydryl compound of glutathion and cysteine etc. has distinctive undesirable abnormal smells from the patient, and percutaneous absorption aspect also has problems in addition, and these glycocides and derivant also polarity are high, so be difficult to use as the preparation composition of cosmetic material.In addition, vitamin C is easily oxidation under aqueous solution state, and existence can not be brought into play the problem of lasting effect.
Therefore, developed in recent years the skin lightening compositions that contains natural herbal extract, but major part presents color, and the upper existing problems of configuration, its effective ingredient is not yet identified, has the problem that can not expect effect same in the time of preparing product for this reason.
In order to address the above problem, those skilled in the art make great efforts to develop the single product that whitening effect is good, safe, have no side effect, and its result has completed the present invention.
The invention provides a kind of dermato-cosmetic composition that contains asiaticoside, its object is that the melanin that not only suppresses to produce in the melanocyte of skin generates, and suppress skin melanism cytokine PGE2 generation secreted in skin keratin formation cell, thereby play whitening effect.
Summary of the invention
For reaching above object, state according to the present invention provides a kind of dermato-cosmetic composition that contains asiaticoside.
State provides a kind of dermato-cosmetic composition that contains asiaticoside as cosmetic material compositions, pharmaceutics compositions or food compositions according to another preferred.
According to the state of another one more of the present invention, the invention provides and making the skin method of whitening more, its method is described dermato-cosmetic composition to be carried out on Dermatology to administration on the individuality in allowed band.
According to the present invention, asiaticoside not only suppresses the generation of secreted skin melanism cytokine in keratinocyte, but also melanin in check melanin cell generates, thereby has skin-whitening effect.In addition, in human safety test, any toxicity or stimulation not occurring, made it to distinguish that asiaticoside is the safe material that goes for human body.Therefore, asiaticoside of the present invention can be widely used in cosmetic material, medicine or the food etc. of skin-whitening.
Accompanying drawing explanation
Fig. 1 is by the system diagram of the keratinocyte of skin and the common training of melanocyte.
Fig. 2 is on the artificial skin of irradiation ultraviolet radiation, and after asiaticoside 0.5% is processed, the melanin of artificial skin scatters the comparison diagram with matched group.
The specific embodiment
The effective ingredient asiaticoside (Madecassoside) comprising in compositions of the present invention is the material that is mainly present in triterpenoid (triterpenoid) series of Herba Centellae (Centella asiatica), and generation, improvement treatment wound healing that it can reduce keloid are well-known.There is the structure of following Chemical formula 1.
[Chemical formula 1]
Described asiaticoside is natural source (natural source), can extract and form from Herba Centellae (Centella asiatica[L.]).For example, utilize multiple extracting method well known in the art can obtain asiaticoside from described natural source.Be preferably, (a) mixed solvent, (d) acetone, (e) ethyl acetate, (f) chloroform, (g) 1,3 butylene glycol or the mixture by them of the anhydrous or moisture lower alcohol (methanol, ethanol, propane, butane etc.) of water, (b) carbon number 1-4, (c) described rudimentary alcohol and water can be used as extraction solvent and obtain asiaticoside.
More preferably, can utilize the extraction with aqueous solution asiaticoside of methanol and ethanol, most preferably be, can utilize butanols extraction with aqueous solution asiaticoside.
On the other hand, asiaticoside of the present invention not only utilizes above-mentioned solvent extraction, but also can pass through other solvent extractions, and can obtain the extract of same effect, and this is self-explantory for a person skilled in the art.Those skilled in the art are except extracting solvent, and required concrete extraction conditions (temperature, time etc.), can independently select and adjust according to the character of described extraction solvent.
In addition, extract of the present invention not only extracts by described extraction solvent, but also comprises by conventional tablet engineering and extracting.For example, the ultrafilter membrane that utilization has certain molecular weight cut-off value separates, according to chromatography, (for separating and make according to size, electric charge, hydrophobicity, affinity) separates etc., and in addition, extract of the present invention also comprises by implementing multiple agent method and obtains graduation.In addition, the asiaticoside using in the present invention can be the product (for example: the asiaticoside of Chromadex company) through chemosynthesis making or business making.
Asiaticoside of the present invention is with respect to the existing vitamin C (vitamin C) that is applicable to skin-lightening cosmetic raw material, and the inhibition that in its whitening screening substances, normally used intracellular tyrosine enzymatic activity and melanin generate is superior more than 50%.In addition, asiaticoside of the present invention has the effect that in the keratinocyte that suppresses skin, secreted skin melanism cytokine PGE2 generates.Therefore, asiaticoside of the present invention is applied to melanocyte and the keratinocyte of all skins, has melanin in the cell of inhibition and generates, and prevent the Pigmented effect of skin.
On the other hand, the asiaticoside of Chemical formula 1 of the present invention is based on art technology level, conventionally the derivant that the interpolation sub stituent of implementing by this area or displacement reaction obtain comprises the derivant that plays whitening effect through suppressing skin melanism inductive substance, and this it will be apparent to those skilled in the art that.
In lightening compositions of the present invention, described asiaticoside comprises approximately 0.0001 % by weight to approximately 15 % by weight content in composition total weight.When described asiaticoside content is less than 0.0001 % by weight, whitening effect is faint, and while exceeding 15 % by weight, owing to increasing content, to increase effect meagre, and existence can not be guaranteed the problem of the stability in dosage form.More preferably, asiaticoside comprises 0.001 % by weight to 10 % by weight in composition total weight.
As preferred embodiment, the cosmetic material compositions that dermato-cosmetic composition of the present invention can be object for skin whitening.
Cosmetic material compositions of the present invention, as effective ingredient, beyond hydroxyl-removal asiaticoside, comprises the composition that is conventionally applied to cosmetic material compositions.For example, can comprise conventional auxiliary agent and or the carriers such as antioxidant, stabilizing agent, lytic agent, vitamin, pigment and spice.And described cosmetic material compositions, in order to promote skin-whitening effect, can also comprise the material that promotes that skin absorbs in addition.
Cosmetic material compositions of the present invention also can be prepared any dosage form that this area makes conventionally.For example, can be prepared as solution, suspension, emulsion, paste, gel, cream, emulsion, powder, soap, contain abluent, oil preparation, muffin, foundation emulsion, wax foundation cream and the spray etc. of surfactant, but be not limited to this.More specifically, can be prepared as the dosage form of soft astringent, nutrition astringent, nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing milk, makeup removing water, facial film, spray or powdery.
When dosage form of the present invention is paste, cream or gel, can utilize animal oil, vegetable oil, wax, paraffin, starch, cellulose derivative, Polyethylene Glycol, silicone, bentonite, Silicon stone, Talcum or zinc oxide etc. as carrier components.
When dosage form of the present invention is powdery or spray, lactose, Talcum, Silicon stone, aluminium hydroxide, calcium silicates or Silon can be utilized as carrier components, especially when spray, the promoter such as HCFC, propane, butanols or dimethyl ether can also be comprised.
When dosage form of the present invention is solution or emulsion, utilize solvent, thawing agent or opacifiers as carrier components, for example, water, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzylalcohol, benzyl benzoate, propylene glycol, 1,3 butylene glycol oil, fatty acid glyceride, Polyethylene Glycol or sorbitan fatty acid ester.
When dosage form of the present invention is suspension, can utilize as the liquid diluent of water, ethanol or propylene glycol as carrier components, as the suspending agent of ethoxylated isostearyl alcohol, polyoxyethylene sorbitan ester, polyoxyethylene sorbitan ester, small crystalline cellulose, magaldrate, amargosite, agar or Tragacanth etc.
Dosage form of the present invention is, while containing the abluent of surfactant, can utilize analiphatic sulphur acid alcohol, aliphatic sulphuric acid alcohol ether, sulfosuccinic acid monoesters, isethionic acid, imidazoles woods derivant, N-methyltaurine sodium, sarcosinate, fatty acid amide ether sulfate, alkylamide propyl-betaine, aliphatic alcohol, fatty glyceride, fatty diglycollic amide, vegetable oil, lanolin derivative or ethoxylated glycerol fatty acid ester etc. as carrier components.
In cosmetic material compositions of the present invention, the content of asiaticoside can comprise 0.0001 % by weight to 15 % by weight of composition total weight, is preferably 0.001 % by weight to 10 % by weight.
Another preferred embodiment, it is object pharmaceutics compositions that dermato-cosmetic composition of the present invention can be used for skin-whitening.
In pharmaceutics compositions of the present invention, the content of asiaticoside can comprise 0.0001 % by weight to 15 % by weight of composition total weight, is preferably 0.001 % by weight to 10 % by weight.
Pharmaceutics compositions of the present invention can also comprise and is conventionally applied to optimum carrier, excipient and the diluent of preparing pharmaceutics compositions.
The dosage form of pharmaceutics compositions of the present invention is according to conventional method in pharmaceutics field, in can pharmaceutics field, the free position of preparation be carried out dosage form, for example, can make the plaster type dosage forms such as powder, granule, lozenge, capsule, suspension, Emulsion, syrup, gaseous solvents, the external preparation such as ointment, cream, sits agent and sterilizing injection etc.
Pharmaceutics compositions of the present invention can in the mammals such as rat, mice, domestic animal, people, lung meridian mouth, per os etc. can number of ways administrations.For example, can pass through per os, rectum or vein, muscle, subcutaneous, intrauterine (intracerebroventricular) drug administration by injection in film or cerebrovascular.Now, non-per os path is preferably percutaneous dosing, and it most preferably is partial smearing.
The dosage of described preparation is according to patient's age, sex, body weight, symptom, and its medication is different, but during for external preparation, smears 1.0 every day to 3.0ml, and smear continuously more than 1 month every day for 1 to 5 time.In addition, when per os dosage form, different according to patient's age, sex, body weight, can take 0.1 to 100 ㎎/㎏ at every turn, every day 1 is to repeatedly.Just, can be according to increase and decrease dosages such as administration path, the order of severity, sex, body weight, ages.Thereby described dosage where face in office, does not limit scope of the present invention.
Another preferred embodiment, skin lightening compositions of the present invention is applicable to the functional food composite take skin-whitening as object.
Food compositions of the present invention, as effective ingredient, not only comprises asiaticoside, also comprises the composition conventionally adding while preparing food, for example, and protein, carbohydrate, fat, nutrient, flavoring agent and flavouring agent.Described carbohydrate is as monosaccharide, for example, and glucose, fructose etc., disaccharide, such as maltose, sucrose, oligosaccharide etc.; And polysaccharide, for example, glucide, cyclodextrin etc. are used the sugar alcohols such as sugar and xylitol, Sorbitol, erythritol conventionally.Can use natural flavours (thaumatin, stevioside derivant, [such as content rebaudioside-A, glycyrrhizin etc.]) and synthetic flavouring agent (glucide, aspartame etc.) as flavouring agent.
For example, when food compositions of the present invention is prepared as beverage, except asiaticoside of the present invention, can also comprise citric acid, liquid fructose, Saccharum Sinensis Roxb., glucose, acetic acid, malic acid, fruit juice, Cortex Eucommiae derivant, Fructus Jujubae derivant, Radix Glycyrrhizae derivant etc.
In specific embodiments of the invention, asiaticoside proves that to skin accumulative total irritant test result asiaticoside, as natural materials, is a kind of harmless material.Therefore, asiaticoside of the present invention does not almost have toxicity or side effect, so, even when life-time service, also can relievedly use, especially can safe and applicable in cosmetic material compositions as above, pharmaceutics compositions and food compositions.
On the other hand, the present invention also provides a kind of method of skin whitening, and the skin lightening compositions that contains asiaticoside is carried out to administration to individuality in Dermatology tolerance band.
' individuality ' of the present invention refers to and improved or prevent people or the mammal due to skin melanism or ultraviolet, hormone or the genetic cutaneous pigmentation causing, as Canis familiaris L., pig, horse, cattle etc.
Pull of the present invention refers to arbitrarily suitable method to the individual compositions of injecting regulation, compositions of the present invention can per os or non-per os carry out administration, administration path can be any general path of tissue of can achieving the goal.In addition, compositions of the present invention is the destination object of active substance, for example, can carry out administration by any device that can move to cell.
According to method of the present invention, the skin melanism of described individuality or cutaneous pigmentation disease can by by skin lightening compositions of the present invention to carrying out administration in individuality, can reach the objects such as improvement, improvement, prevention or treatment.The technical staff of general knowledge known in this field is understandable by described content and following enforcement.
[specific embodiment]
Illustrate in greater detail the present invention below by embodiment.These embodiment are only used to be described more specifically the present invention, and invention which is intended to be protected is not limited to these embodiment, and the technical staff of general knowledge known in this field is stated hereby.
Embodiment 1: measure the melanin generation that asiaticoside suppresses B16 melanoma cells
Utilize the pigment cell (B16 melanoma cells) of rat to measure the effect being generated by asiaticoside check melanin, the effect that the check melanin being produced by vitamin C that this mensuration effect is generated with known check melanin generates is compared.
Be specially, on 6 orifice plates with every hole 1 10
5mice Inoculated melanoma (B-16 F10) cell, in the DMEM culture medium of FBS (too Ox blood serum) that contains 10%, at 5%CO
2with at 37 ℃, cultivate into cell and adhere to approximately 80% in the bottom in hole.After cultivation, remove culture medium, the culture medium of asiaticoside and vitamin C dilution debita spissitudo is processed to 5%CO
2, cultivate 3 days at 37 ℃.The concentration range of asiaticoside is determined is not having Cytotoxic 1uM, 10uM, 50uM.Will remove the PBS(phosphated buffer saline for cell of culture medium) clean, by it by trypsin treatment and reclaim cell.The cell being recovered utilizes hematimeter (hematocytometer) to measure after cell number, by each process element, the cell number of equivalent amount is put into 2mL pipe, with 5,000 to 10,000rpm centrifugalize 10 minutes, then removes supernatant and is precipitated.After this cell precipitation is dried at 60 ℃, add the 1M sodium hydroxide solution 100 μ l that contain 10% DMSO to obtain intracellular melanin in 60 ℃ of temperature chambers.Use this liquid, measure absorbance by microplate reader (microplate reader) at 490nm, obtain the melanin amount of each fixed number cell.Its result is as shown in table 1.
[table 1]
As shown in Table 1 above, in same concentration for the treatment of, showed the asiaticoside melanin generating suppression more superior than vitamin C.
Embodiment 2: measure asiaticoside and suppress the effect that in human body melanocyte, melanin generates
Utilize human body melanocyte (melanocyte) to measure the effect that asiaticoside check melanin generates, the vitamin C that this mensuration effect is generated with known check melanin suppresses the effect of intracellular tyrosine enzymatic activity and compares.
Particularly, buy and suppress human epidermal melanocyte (Human epidermal melanocyte[comes from neonate]) and cultivate comprising the culture medium 254 of buying human body melanocyte growth additive (Human Melanocyte Growth supplement) from above-mentioned company from Cascade Biologics company.On 6 orifice plates with every hole 1 10
5after inoculation human epidermal melanocyte (Human epidermal melanocytes), at 5%CO
2with at 37 ℃, cultivate into cell and adhere to approximately more than 80% in the bottom in hole.After cultivation, remove culture medium, sample is diluted to the culture medium of debita spissitudo and process, 5%CO
2, culture medium culturing of displacement in two days 5 days at 37 ℃.The concentration range of asiaticoside is determined at 1uM, 10uM.Will remove after end process the PBS(phosphated buffer saline for cell of culture medium) clean, by it by trypsin treatment and reclaim cell.The cell being recovered utilizes hematimeter (hematocytometer) to measure cell number, by each process element, the cell number of equivalent amount is put into 2mL pipe, then with 5,000 to 10,000rpm centrifugalize 10 minutes, then removes supernatant and be precipitated.After this cell precipitation is dried at 60 ℃, add the 1M sodium hydroxide solution 100 μ l that contain 10%DMSO to obtain intracellular melanin in 60 ℃ of temperature chambers.Use this liquid, measure absorbance by microplate reader (microplate reader) at 490nm, obtain the melanin amount of each fixed number cell.Its result is as shown in table 2.
[table 2]
| Style | Concentration for the treatment of (uM) | Melanin generating suppression (%) |
| Vitamin C | 1 | 0 |
| Vitamin C | 5 | 2 |
| Vitamin C | 10 | 6 |
| Asiaticoside | 1 | 19 |
| Asiaticoside | 5 | 28 |
| Asiaticoside | 10 | 51 |
As described in above-mentioned table 2, in unified concentration for the treatment of, show the suppression ratio that in the asiaticoside cell more superior with respect to vitamin C, melanin generates.
Embodiment 3: measure the effect suppressing by the PGE2 generation of ultraviolet induction keratinocyte
Utilize the effect of human keratinous formation raji cell assay Raji asiaticoside check melanin inducing cell factor PGE2.
Particularly, buy and suppress human epidermal melanocyte (Human epidermal melanocyte[comes from neonate]) and cultivate comprising the EpiLife culture medium of buying human body melanocyte growth additive (Human Melanocyte Growth supplement) from above-mentioned company from Cascade Biologics company.On 6 orifice plates with every hole 1 10
5inoculation keratinocyte, at 5%CO
2with at 37 ℃, cultivate into cell and adhere to approximately more than 80% in the bottom in hole.After cultivation, remove culture medium, sample is diluted to the culture medium of debita spissitudo and process, 5%CO
2, carry out after pre-treatment in 24 hours at 37 ℃, then irradiate artificial ultraviolet (UVB 20mJ) after using PBS (phosphated buffer saline) displacement.Finish, after irradiation ultraviolet radiation, to remove PBS, exchange asiaticoside is pressed the keratinocyte culture fluid of concentration processing, then cultivates 24 hours.Assemble cell culture fluid, utilize business PGE2 test kit (R & D) to measure endocrine culture fluid PGE2.Its result is as shown in table 3.
[table 3]
As described in above-mentioned table 3, can confirm the generation that is increased PGE2 by ultraviolet radiation, reduce concentration for the treatment of by asiaticoside.
Embodiment 4: keratinocyte-melanocyte is measured the effect that inhibition is generated by ultraviolet induction melanin under co-cultivation facility
Shown in Fig. 1, measured the effect generating by asiaticoside check melanin, this effect utilization keratinocyte and melanocyte co-cultivation facility.Be specially, melanocyte is seeded on 12 holes, keratinocyte is seeded on micropore (Millicell).Confirm after the glial cell that is seeded in micropore adheres to, asiaticoside to be processed by concentration.After removing culture fluid PBS displacement after 1 day, irradiate UVB.Then,, after asiaticoside is replaced again by the culture fluid of concentration processing, micropore is placed on 12 holes of inoculation melanocyte.This co-cultivation facility, due to by the filter membrane mobile culture fluid mutually of micropore, can be observed the mutual variation of material in culture fluid.
After end process, remove culture medium, utilize PBS (phosphated buffer saline) to clean, by trypsin treatment and reclaim cell.Utilize hematimeter (hematocytometer) to measure after cell number in the cell of recovery, by each process element, the cell number of equivalent amount is put into 2mL pipe, after 10 minutes, remove supernatant with 5,000 to 10,000rpm centrifugalize and be precipitated.Make this cell precipitation 60 ℃ dry after, add the 1M sodium hydroxide solution that contains 10% DMSO, in the temperature chamber of 60 ℃, obtain intracellular melanin.Use this liquid, measure absorbance by microplate reader (microplate reader) at 490nm, obtain the melanin amount of each fixed number cell.This result is as shown in table 4.
[table 4]
| Style | Concentration for the treatment of (uM) | Melanin generating suppression (%) |
| Asiaticoside | 10 | 19 |
| Asiaticoside | 50 | 28 |
| Asiaticoside | 100 | 51 |
As shown in Table 4 above, its result shows generating according to control of the concentration melanin of asiaticoside.
Embodiment 5: measure whitening effect in artificial skin
Artificial skin uses the MEL-300-B (MaTek.USA) for buying, and artificial skin culture fluid EPI-100-LLMM (MaTek.USA) displacement in every 3 days that comprises following table 5 content has once been processed 2 weeks.Before sample displacement, irradiate ultraviolet with UVB 50mJ condition.After the component that processing finishes is fixing in 10% formaldehyde, thinly-sliced after recycling paraffin embeds, carry out Fontana-Masson dyeing with the special melanin method of dying.After this, spread melanin amount by the every certain area of Image-pro plus software (MediaCybernetics, U.S.A).Its result is as shown in table 5 and Fig. 2.
[table 5]
As shown in table 5, its result shows obviously to have reduced melanin distributed density in asiaticoside processed group according to concentration.
Embodiment 6: as peroral administration whitening effect
When irradiation ultraviolet radiation, due to promotion skin melanism process that can be artificial, after animal experiment middle-ultraviolet lamp irradiates, sample per os or percutaneous dosing are measured to its effect, can confirm thus actual whitening effect.
Therefore, in the present embodiment, used known to brown Cavia porcellus (the Tortoiseshell guinea pigs of the similar ultraviolet chromogenesis calmness of people; Brown guinea pigs), measure the whitening effect of asiaticoside.The Cavia porcellus of 6-7 one full year of life is divided into 8 of every conventional groups, and UV matched group, UV/ asiaticoside administration group, wait upon at duration of test.Conventional group gives 0.5ml normal saline with matched group per os, and UV/ asiaticoside administration group is take solid type powder as benchmark, and every body weight Kg utilizes administrable injection that 100mg asiaticoside is mixed in to 0.5ml saline solution per os to carry out administration.
The administration time limit is totally 4 weeks, and day same time has been carried out administration on every Fridays.When oral administration 2 weeks, irradiate (total irradiation energy=1350mJ/cm to UV matched group and asiaticoside administration group
2) SE lamp (wavelength is 290-320nm, Toshiba).There is pigmentation phenomenon after 2 or 3 weeks in ultraviolet radiation, after approximately 2 weeks, has measured skin pigment variation.
Use colour difference meter (Minolta CR2002 chroma meter) to judge black and white degree and the effect of measuring skin, its result is as shown in table 6.In expression color, use L
*a
*b
*colour system, in the present embodiment L
*value is index.Proofread and correct L with standard white plate
*value, L
*value is measured more than 5 times repeatedly at a position, thereby has realized impartial pigmentation.
[table 6]
| Sample | Whitening effect (L *) |
| UV matched group | 0.52?0.09 |
| UV/ asiaticoside | 0.85?0.08 |
As shown in Table 6 above, can observe asiaticoside administration group and there is higher whitening effect (L with respect to the UV/ matched group that there is no administration
*be worth larger skin-color brighter).
Embodiment 7: confirm the test of asiaticoside to human body skin safety
The preparation of the skin preparations for extenal use that 7-1. comprises asiaticoside
According to described embodiment, in order to confirm to distinguish asiaticoside for the whitening effect excellence safe coefficient to human body skin, respectively prepare and contained asiaticoside and ascorbic skin preparations for extenal use, carry out the confirmatory experiment of cutaneous safety.
According to the component content of following table 7, prepare and respectively contained asiaticoside and ascorbic skin preparations for extenal use.In following table 7, matched group is the skin preparations for extenal use that does not contain tryrosinase co-inhibitor, and test group 1 and test group 2 are to contain separately asiaticoside and ascorbic skin preparations for extenal use.
In order to prepare skin preparations for extenal use, mix purify waste water, propylene glycol and butanediol at 70 ℃, dissolve (aqueous portion), at 70 ℃, dissolve the residual components (oil phase part) except described 3 kinds of compositions and trimethanolamine.In aqueous portion, add described oil phase part, stir and carry out 1 emulsifying with homogenizer (Japanese Tokushu Kika company produce), finally added therein trimethanolamine.Remove after the foaming generating in mixed liquor, at room temperature carry out cooling and prepared skin preparations for extenal use.
[table 7]
7-2. skin accumulative total irritant test
Use each skin preparations for extenal use of preparing in described embodiment 7-1, the next day of carrying out at the upper arm position of 30 health adults totally 9 times, the accumulative total patch of each 24 hours, has measured asiaticoside and whether skin has been caused to stimulation.
Patch method is to have utilized Finn-Chamber (the Finn chamber of Finland Epitest Ltd).In cell, drip separately after described skin preparations for extenal use 15ul at every turn and carried out patch.Utilize following empirical formula 1 to mark the extent of reaction at every turn occurring on skin, its result as described in Table 8.
[empirical formula 1]
Average response degree=[[index of Response x degree of reaction/be verified total number of persons x highest score (4 points)] x100] ÷ checks number of times (9 times)
Now, in degree of reaction, give pen 1 point ,+be 2 points, ++ be the mark of 4 points, average response degree, lower than 3 o'clock, is judged to be safe compositions.
[table 8]
In described table 8, in test group 1, about ,+, ++ relative number be respectively 1,0,0, in addition do not occur reaction.Be calculated as [(1x2)/(20x4)] x100/9=0.13 according to empirical formula as implied above, average response degree is 0.37, lower than 3, is judged to be safe compositions.Thereby the skin preparations for extenal use district that comprises asiaticoside (test group 1), obviously not in matched group or contain ascorbic skin preparations for extenal use, does not show the state that accumulative total stimulates, and is judged to be the material to human-body safety.
formulation example 1: cosmetic material preparation
formulation example 2: medicine and pharmacology preparation
1. the preparation of powder
Asiaticoside 2g
Lactose 1g
After mixing described composition, filling is prepared powder in airtight cloth.
2. the preparation of tablet
Asiaticoside 100 ㎎
Corn starch 100 ㎎
Lactose 100 ㎎
Dolomol 2 ㎎
Mix after mentioned component, method for preparing tablet thereof is routinely played ingot and is prepared tablet.
3. the preparation of capsule
Asiaticoside 100 ㎎
Corn starch 100 ㎎
Lactose 100 ㎎
Dolomol 2 ㎎
Mix after mentioned component, the preparation method filling of capsule is prepared capsule in gel capsule routinely.
Claims (5)
1. a skin lightening compositions, it comprises asiaticoside as effective ingredient.
2. compositions according to claim 1, is characterized in that: the melanin that described asiaticoside effectively suppresses dermal melanin cell generates, and effectively suppresses skin melanism cytokine PGE2 secreted in the keratinocyte of skin.
3. compositions according to claim 1 and 2, is characterized in that: the content of described asiaticoside accounts for 0.0001 % by weight to 15 % by weight of composition total weight.
4. compositions according to claim 1, is characterized in that: described compositions is to be used in cosmetic material, medical material or the food take skin-whitening as object.
5. compositions according to claim 4, is characterized in that: in the situation that described compositions is cosmetic material, and dosage form composed of the following components; For example, by solution, suspension, emulsion, paste, gel, cream, emulsion, powder, soap, form containing abluent, cleaning agent, oil preparation, muffin, foundation emulsion, wax foundation cream and the spray of surfactant.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/KR2012/005211 WO2014003224A1 (en) | 2012-06-29 | 2012-06-29 | Compositions for whitening skin comprising madecassoside |
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| Publication Number | Publication Date |
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| CN103796631A true CN103796631A (en) | 2014-05-14 |
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| CN201280021188.7A Pending CN103796631A (en) | 2012-06-29 | 2012-06-29 | Compositions for whitening skin comprising madecassoside |
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| Country | Link |
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| JP (1) | JP2014520166A (en) |
| CN (1) | CN103796631A (en) |
| WO (1) | WO2014003224A1 (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104434618A (en) * | 2014-08-28 | 2015-03-25 | 济南臻雅化妆品有限公司 | Nine-herb cream with effects of whitening and removing freckle marks and preparation method thereof |
| CN112957764A (en) * | 2021-01-29 | 2021-06-15 | 三益创价生物科技(深圳)有限公司 | Method for extracting asiaticoside-rich composition from centella asiatica |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR102371416B1 (en) * | 2015-09-30 | 2022-03-08 | (주)아모레퍼시픽 | Method for making senescent melanocyte cells, senescent cells therefrom and method for screening anti-aging material using the same |
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| CA2628694A1 (en) * | 2005-11-09 | 2007-05-18 | Bayer Consumer Care Ag | Use of compounds from centella asiatica |
| JP2007297367A (en) * | 2005-12-15 | 2007-11-15 | Yomeishu Seizo Co Ltd | Composition for controlling elevation of blood pressure, medicine and functional food |
| JP2011057634A (en) * | 2009-09-11 | 2011-03-24 | Rohto Pharmaceutical Co Ltd | Skin-lightening composition |
| KR101064904B1 (en) * | 2011-01-21 | 2011-09-16 | 주식회사 씨앤피차앤박화장품 | Cosmetic composition for improving acne comprising natural extracts |
| KR101245563B1 (en) * | 2011-03-16 | 2013-03-21 | 바이오스펙트럼 주식회사 | Agents for skin whitening containing Madecassoside |
| JP6026785B2 (en) * | 2011-10-31 | 2016-11-16 | 富士フイルム株式会社 | Aqueous composition |
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- 2012-06-29 CN CN201280021188.7A patent/CN103796631A/en active Pending
- 2012-06-29 JP JP2014522735A patent/JP2014520166A/en active Pending
- 2012-06-29 WO PCT/KR2012/005211 patent/WO2014003224A1/en active Application Filing
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| CN1194154A (en) * | 1997-03-24 | 1998-09-30 | 东国制药株式会社 | Asiaticoside of self asiatic and hydroxy-asiaticoside and carboxy-asiaticoside water soluble extract and its separating method |
| CN101596152A (en) * | 2008-03-28 | 2009-12-09 | 莱雅公司 | The compositions of the combination of hydroxyl asiaticoside and/or terminoloside and arginine and/or its salt and/or its derivant and purposes |
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| CN104434618A (en) * | 2014-08-28 | 2015-03-25 | 济南臻雅化妆品有限公司 | Nine-herb cream with effects of whitening and removing freckle marks and preparation method thereof |
| CN112957764A (en) * | 2021-01-29 | 2021-06-15 | 三益创价生物科技(深圳)有限公司 | Method for extracting asiaticoside-rich composition from centella asiatica |
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| WO2014003224A1 (en) | 2014-01-03 |
| JP2014520166A (en) | 2014-08-21 |
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