CN105663595B - A kind of melanin inhibitor, skin care compositions and its application - Google Patents
A kind of melanin inhibitor, skin care compositions and its application Download PDFInfo
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- CN105663595B CN105663595B CN201610109586.4A CN201610109586A CN105663595B CN 105663595 B CN105663595 B CN 105663595B CN 201610109586 A CN201610109586 A CN 201610109586A CN 105663595 B CN105663595 B CN 105663595B
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
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- 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
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/34—Alcohols
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- 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
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/34—Alcohols
- A61K8/345—Alcohols containing more than one hydroxy group
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- 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
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/37—Esters of carboxylic acids
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- 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
- A61K8/40—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing nitrogen
- A61K8/44—Aminocarboxylic acids or derivatives thereof, e.g. aminocarboxylic acids containing sulfur; Salts; Esters or N-acylated derivatives thereof
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
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- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
- A61K8/4973—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with oxygen as the only hetero atom
- A61K8/498—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with oxygen as the only hetero atom having 6-membered rings or their condensed derivatives, e.g. coumarin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- 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
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
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- A61K8/00—Cosmetics or similar toiletry preparations
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- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/67—Vitamins
- A61K8/673—Vitamin B group
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- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
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- A61K8/676—Ascorbic acid, i.e. vitamin C
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- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/73—Polysaccharides
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- A61K8/00—Cosmetics or similar toiletry preparations
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- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/81—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds obtained by reactions involving only carbon-to-carbon unsaturated bonds
- A61K8/8141—Compositions of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by only one carboxyl radical, or of salts, anhydrides, esters, amides, imides or nitriles thereof; Compositions of derivatives of such polymers
- A61K8/8147—Homopolymers or copolymers of acids; Metal or ammonium salts thereof, e.g. crotonic acid, (meth)acrylic acid; Compositions of derivatives of such polymers
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
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- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/84—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds obtained by reactions otherwise than those involving only carbon-carbon unsaturated bonds
- A61K8/86—Polyethers
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- A—HUMAN NECESSITIES
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- 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
- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/84—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds obtained by reactions otherwise than those involving only carbon-carbon unsaturated bonds
- A61K8/89—Polysiloxanes
- A61K8/891—Polysiloxanes saturated, e.g. dimethicone, phenyl trimethicone, C24-C28 methicone or stearyl dimethicone
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
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- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
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- 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
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- 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
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
Abstract
The present invention relates to a kind of melanin inhibitor, skin care compositions and its applications.The melanin inhibitor is dragon's blood extract, can be as tyrosinase inhibitor for inhibiting melanin production with good tyrosinase effect and percutaneous absorbability；The external skin-care composition contains dragon's blood extract and optional bamboo leaf flavone, and bamboo leaf flavone has preferable antioxidation to Resina Draconis, is conducive to the stabilization of Resina Draconis.It can produce preferable synergistic effect after dragon's blood extract is compounded with bamboo leaf flavone, made external skin-care composition is significantly greater than dragon's blood extract single dose to the inhibitory effect of melanin production.Using skin care made of melanin inhibitor or topical composition of the invention, tyrosine activity can be inhibited, reduce melanin production, effectively prevent skin of face melanin deposition and color spot generates.
Description
Technical field
The invention belongs to Chinese medicines and cosmetic technical field, and in particular to a kind of melanin inhibitor, skin care compositions and
It is applied.
Background technique
Tyrosinase is widely present in people, in animal and plant, is the key enzyme that B16 cell is participated in organism.?
In melanocyte, tyrosine is oxidized to DOPA and DOPA quinone under tyrosinase effect, rearranged generation dopachrome, then
It is polymerized to 5,6- dihydroxy indole by series reaction, ultimately forms melanin.Tyrosinase has the above process a variety of
Catalytic activity can be used as tyrosine hydroxylase, dopa-oxidase, can also be used as 5,6- dihydroxy indole oxidizing ferment；It is lived
Property can determine the speed and yield of melanin production, inhibit its activity to can control Melanin productions.It is widely used at present
Tyrosinase inhibitor be mainly that kojic acid or arbutin are deposited although both substance tyrosinase inhibitory activities are good
Stability it is poor, be difficult to the problems such as absorbing.
Therefore, presently, there are the problem of be to research and develop that a kind of tyrosinase inhibitory activity is good, and stability is good, and is easy to
The melanin inhibitor of absorption.
Summary of the invention
An object of the present invention is to provide a kind of application of dragon's blood extract as tyrosinase inhibitor, i.e., by it
As melanin inhibitor.The second object of the present invention is to provide a kind of external skin-care composition containing dragon's blood extract.
The third object of the present invention is to provide a kind of dragon's blood extract or the external skin-care composition containing dragon's blood extract is protecting
Application in skin product.
For this purpose, first aspect present invention provides a kind of melanin inhibitor, it is dragon's blood extract.
Second aspect of the present invention provides a kind of for inhibiting the external skin-care composition of melanin, mentions containing Resina Draconis
Take object.
According to the present invention, in the composition, total weight of the content of the dragon's blood extract based on composition
For 90wt%-95wt%.
According to the present invention, the composition also contains bamboo leaf flavone.
In some specific preferred embodiments of the invention, in the composition, the total weight based on composition,
Content≤10wt% of bamboo leaf flavone described in 5wt% <.
Third aspect present invention provides a kind of for inhibiting the external-use skin care product of melanin, contains such as the present invention the
Melanin inhibitor described in one side or external skin-care composition as described in respect of the second aspect of the invention.
Other embodiments according to the present invention, in the external-use skin care product, the dosage of the melanin inhibitor
For 0.25wt%-1wt%.
According to some embodiments of the present invention, in the external-use skin care product, the dosage of the external skin-care composition
For 0.26wt-1.1wt%.
According to the present invention, the external-use skin care product further include moisturizer, it is oil components, antioxidant, thickener, ultraviolet
One of light absorbers, surfactant, water composition and water are a variety of.
In some embodiments of the invention, the external-use skin care product with frost, lotion, facial mask and is washed in baths extremely
A kind of few form exists.
Fourth aspect present invention provides a kind of for inhibiting the face cream of melanin, is based on face cream total weight, the face
Frost includes:
According to the present invention, the face cream also includes one in pawpaw sulfydryl enzyme, Bilberry fruit P.E and sodium ascorbyl phosphate
Kind is several；It is preferably based on face cream total weight, dosage≤1wt% of pawpaw sulfydryl enzyme, further preferably 0.5wt%-
1wt%；Dosage≤1wt% of Bilberry fruit P.E, further preferably 0.5wt%-1wt%；The dosage of sodium ascorbyl phosphate
≤ 4wt%, further preferably 2wt%-4wt%.
Detailed description of the invention
Fig. 1 shows the dragon's blood extract of various concentration in the present invention for inhibitory activity against tyrosinase.
Fig. 2 is dragon's blood extract high-efficient liquid phase chromatogram in the present invention.
Fig. 3 is 30min Transdermal absorption ingredient high-efficient liquid phase chromatogram in the present invention.
Fig. 4 is 60min Transdermal absorption ingredient high-efficient liquid phase chromatogram in the present invention.
Fig. 5 is 90min Transdermal absorption ingredient high-efficient liquid phase chromatogram in the present invention.
Specific embodiment
To keep the present invention easier to understand, below in conjunction with embodiment, the present invention will be described in detail, these embodiments are only
Serve illustrative, it is not limited to application range of the invention, unmentioned specific experiment method in the following example, usually
It is carried out according to routine experiment method.
Resina Draconis is Liliaceae dracaena swordleaf dragon tree Dracaena cochinchinensis (Lour.)
The resin that the xylem of S.C.Chen extracts has promoting blood circulation to remove blood stasis, analgesic, hemostasis, myogenic sore and other effects, main chemical compositions
Including flavones, volatile oil, phenols etc..Mainly application includes anti-inflammatory, analgesic, hypoglycemic etc. to Resina Draconis at present, in treatment angiocarpy
Disease, injure outside it is curative for effect in terms of gynaecologic hemorrhage, but there is not yet about Resina Draconis and its extract as tyrosinase suppression
The report of formulation application.And the present inventor has found that there is preferable tyrosinase to inhibit to make for Resina Draconis and its extract after study
With can be as tyrosinase inhibitor for inhibiting melanin production.
Therefore, melanin inhibitor involved in first aspect present invention is dragon's blood extract, with preferable junket
Propylhomoserin enzyme effect, can be as tyrosinase inhibitor for inhibiting melanin production.Therefore, involved by first aspect present invention
Melanin inhibitor be application that dragon's blood extract can be understood as dragon's blood extract as tyrosinase inhibitor, i.e.,
It is used as melanin inhibitor.
The present inventor also found that bamboo leaf flavone itself, which does not have tyrosinase, directly to be inhibited to make after study
With, but bamboo leaf flavone has preferable antioxidation to dragon's blood extract, is conducive to dragon's blood extract
Stablize.It can produce preferable synergistic effect, made external application after dragon's blood extract is compounded with bamboo leaf flavone
Skin care compositions is significantly greater than dragon's blood extract single dose to the inhibitory effect of melanin production.
For inhibiting the external skin-care composition of melanin involved in second aspect of the present invention, extracted containing Resina Draconis
Object.The composition, which can be understood as one kind, has tyrosinase inhibitory action, and thus further suppresses Melanin productions
External skin-care composition.
In some embodiments of the invention, in the composition, the content of the dragon's blood extract is based on combination
The total weight of object is 90wt%-95wt%.
Some preferred embodiments according to the present invention, the composition also contain bamboo leaf flavone.Some
In embodiment, for example, in the composition, the total weight based on composition, bamboo leaf flavone described in 5wt% <
Content≤10wt%.
For inhibiting the external-use skin care product of melanin involved in third aspect present invention, contain such as the present invention first
Melanin inhibitor described in aspect or external skin-care composition as described in respect of the second aspect of the invention.This can be understood as dragon's blood
Draconis extract or application of the external skin-care composition in skin care containing dragon's blood extract.
In some embodiments of the invention, for example, in the external-use skin care product, based on the total of external-use skin care product
Poidometer, the dosage of the melanin inhibitor are 0.25wt%-1wt%.
In other embodiments of the invention, for example, in the external-use skin care product, based on external-use skin care product
Total weight, the dosage for stating external skin-care composition are 0.26wt-1.1wt%
In the present invention, above-mentioned external-use skin care product can be understood as inhibiting melanin as described in the first aspect of the invention
Agent or external skin-care composition as described in respect of the second aspect of the invention are applied to prepare skin care item, by itself and cosmetics or skin care item
In usual component combination after skin care obtained finished product or semi-finished product.According to certain embodiments of the present invention, described
External-use skin care product further includes moisturizer, oil components, antioxidant, thickener, ultraviolet absorbing agent, surfactant, water
Property one of ingredient and water or a variety of.
The dosage form of above-mentioned external-use skin care product is not particularly limited in the present invention, these skin cares can be with any
Form exists, for example, the external-use skin care product is by frost, lotion, facial mask and presence in the form of at least one of washing baths.
The present invention is not particularly limited the source of the dragon's blood extract, can be commercially available finished product Resina Draconis and mention
Object is taken, is also possible to directly extract the fresh dragon prepared before preparing external skin-care composition or external-use skin care product
Sanguis Draxonis extract, preferably directly extracted before preparing external skin-care composition or external-use skin care product prepare it is fresh
Dragon's blood extract, the extract can be understood as a kind of using Resina Draconis as the mixture of main active.
According to certain embodiments of the present invention, dragon's blood extract the preparation method is as follows:
500g Resina Draconis crude drug is taken, 80 meshes are smashed it through, adds alcohol reflux extraction 2 times of 8 times of volumes 60%, every time
It extracts 2 hours, dregs of a decoction merging filtrate is filtered to remove, 55 DEG C in 55 DEG C of recycling ethyl alcohol on Rotary Evaporators, vacuum oven
Under the conditions of it is dry, remove ethyl alcohol, obtain dragon's blood extract, yield 3.8%.
The method of dragon's blood extract pulverization process is as follows: using Lowtemperaturepulverizer, liquid feeding nitrogen crushes, and pulverizer revolution is
15000-22000r/min, grinding time 10-20min, partial size 200-800 mesh.
In some embodiments of the invention, it is prepared using above-mentioned composition of the present invention a kind of for inhibiting black
The face cream of pigment, as previously mentioned, the composition contains dragon's blood extract and bamboo leaf flavone.To above-mentioned group in the present invention
The adding manner for closing each constituent in object is not particularly limited, and each constituent both can be configured in advance composition and be used for
Preparation suppression black flour frost, when can also be added in the form of single dose, and be added with unit dose form, the amount of each constituent should be in each group
It is formed by ingredient in the range of the amount of composition.
For example, in some embodiments of the invention, being based on face cream total weight, containing in the face cream: 0.25wt%-
1wt%, preferably 0.5wt%-1wt% are more highly preferred to dragon's blood extract, the 4wt%-6wt% of 0.5wt%, preferably 6wt%'s
C12-C15Alkanol benzoic ether, 4wt%-7wt%, the preferably cyclopentasiloxane of 4wt%, 2.5wt%-4wt%, preferably
Cetanol, the 2wt%-3wt% of 2.5wt%, the preferably apricot kernel oil of 2wt%, 2wt%-4wt%, the preferably Solsperse 2000 of 2wt%
Polyethers -2,2wt%-4wt%, Solsperse 2000 polyethers -21, the 2wt%-3wt% of preferably 2wt%, the preferably butanediol of 2wt%,
0.1wt%-0.3wt%, preferably EDTA-2Na, 2wt%-4wt% of 0.1wt%, preferably 3wt%-4wt%, are more highly preferred to
Glycerol, the 0.2wt%-0.8wt% of 3wt%, the preferably xanthan gum of 0.2wt%, 0.35wt%-0.45wt%, preferably
Carbomer, the 1wt%-2wt% of 0.35wt%, the preferably niacinamide of 1wt%, 1wt%-2wt%, the preferably tocopherol of 1wt%
Palm ester, 0.4wt%-0.8wt%, the preferably preservative of 0.8wt%, water and < 1wt%, preferably 0.01wt%-
0.11wt%, further preferred 0.055wt%-0.11wt% are more highly preferred to the bamboo leaf flavone of 0.055wt%.
Test result shows above-mentioned for inhibiting the face cream of melanin because that can inhibit junket with dragon's blood active component
Propylhomoserin activity, reduces melanin production, effectively prevents skin of face melanin deposition and color spot generates.Bamboo leaf flavone sheet
Body does not have direct repression for tyrosinase, compounds with dragon's blood extract by resisting to Resina Draconis in face cream
Oxidation come reduce melanin production and be conducive to keep dragon's blood extract stability, collaboration inhibit melanin production
It is raw.Dragon's blood extract and bamboo leaf flavone are after compounding, and the effect for inhibiting melanin to generate is more compared with exclusive use
Obviously.
In the above-mentioned face cream for inhibiting melanin, other melanin can also be added and inhibit ingredient, such as pawpaw mercapto
One or more of base enzyme, Bilberry fruit P.E and sodium ascorbyl phosphate, and it is based on face cream total weight, pawpaw sulfydryl
Dosage≤1wt% of enzyme, preferably 0.5wt%-1wt%, further preferably 0.5wt%；The dosage of Bilberry fruit P.E≤
1wt%, preferably 0.5wt%-1wt%, further preferably 0.5wt%；Dosage≤4wt% of sodium ascorbyl phosphate, it is excellent
It is selected as 2wt%-4wt%, further preferably 2wt%.
In the present invention, the content surplus of the water in the above-mentioned face cream for inhibiting melanin is based on face cream total weight,
The amount of water is added to 100%.
In the present invention, it is the ingredient being optionally added that " < 1wt% " or "≤1wt% ", which can be understood as the ingredient,.
Heretofore described term " optional " can be understood as certain ingredient and ingredient be added to be alternative, show that the ingredient can
It is added, can also be added without.
Heretofore described " water " word, deionized water, distilled water are referred in the case where being not particularly limited or illustrating
Or ultrapure water.
The present invention is not particularly limited the preparation method for inhibiting the face cream of melanin, can use cosmetics system
It is prepared by the conventional method in standby field, for example, in some embodiments of the invention, the preparation of the face cream for inhibiting melanin
Method the following steps are included:
(1) ingredient is weighed according to above-mentioned formula, Resina Draconis and apricot kernel oil is sufficiently mixed uniformly, cetanol, ring are added
Penta silane of oxygen, C12-C15Alkanol benzoic ether, tocopherol acetate, Solsperse 2000 polyethers -2, Solsperse 2000 polyethers -21, anti-corrosion
Agent is heated to 85 DEG C and sufficiently dissolves and be uniformly mixed, and mixture A is made.
(2) it takes butanediol, EDTA-2Na, glycerol, xanthan gum, carbomer that deionized water mixing is added and is preheated to 85 DEG C, with
Mixture A merges abundant emulsifying, and mixture B is made.
(3) when object B to be mixed is cooled to 50 DEG C or less, sodium ascorbyl phosphate, niacinamide, optional pawpaw mercapto is added
Base enzyme, optional Bilberry fruit P.E, deionized water and essence, after homogeneous mixes, vacuum outgas is then canned, i.e. completion Resina Draconis
The preparation of face cream.
Melanin inhibitor-dragon's blood extract provided by the present invention, with good tyrosinase effect and thoroughly
Skin absorbability, can be as tyrosinase inhibitor for inhibiting melanin production；It is provided by the present invention to be used to inhibit black
The external skin-care composition of element contains dragon's blood extract and optional bamboo leaf flavone, although bamboo leaf flavone
Itself does not have a direct repression for tyrosinase, but bamboo leaf flavone have to Resina Draconis it is preferable anti-oxidant
Effect, is conducive to the stabilization of Resina Draconis.It can produce preferable association after dragon's blood extract is compounded with bamboo leaf flavone
Same-action, made external skin-care composition are significantly greater than dragon's blood extract single dose to the inhibitory effect of melanin production.
Using skin care made of melanin inhibitor or topical composition of the invention, tyrosine activity can be inhibited, reduced black
Pigment generates, and effectively prevents skin of face melanin deposition and color spot generates.
Embodiment
Embodiment 1: fresh dragon's blood extract is prepared.
500g Resina Draconis crude drug is taken, 80 meshes are smashed it through, adds alcohol reflux extraction 2 times of 8 times of volumes 60%, every time
It extracts 2 hours, filters merging filtrate, done under conditions of 55 DEG C in 55 DEG C of recycling ethyl alcohol on Rotary Evaporators, vacuum oven
It is dry, ethyl alcohol is removed, dragon's blood extract, yield 3.8% are obtained.Using Lowtemperaturepulverizer, liquid feeding nitrogen is by dragon's blood extract powder
Broken, pulverizer revolution is 18000r/min, grinding time 15min, partial size 200-800 mesh.
Embodiment 2: measurement of the dragon's blood extract for inhibitory activity against tyrosinase
Experimental principle: tyrosinase can be catalyzed L-3,4 dihydroxyphenylalanine, and product is dopachrome, have maximum at 475nm wavelength
It absorbs, as the generation absorbance of dopachrome is gradually increased, reaction front and back absorbance difference and the ratio of time are tyrosine
The rate of enzymatic reaction, the activity that can embody tyrosinase are strong and weak.
Experimental method: 800 μ g/mL dragon's blood extract solution 1mL use the continuous multiple proportions of pH6.8 phosphate buffer solution
Dilution, acquisition concentration is each 1mL of 800,400,200,100,50,25 μ g/mL extract solution, molten with 0.2%L- DOPA respectively
After mixing, 32 DEG C are incubated for 5 minutes liquid 1mL；Lysyl oxidase solution 1mL is added, continues after being incubated for 10min, in 475nm wavelength
Place's measurement absorbance value.Blank control replaces dragon's blood extract with phosphate buffer.
Dragon's blood extract is shown in Fig. 1 for the inhibiting rate of tyrosinase vigor.
It will be seen from figure 1 that the dragon's blood extract of 200 μ g/mL of concentration can achieve the inhibiting rate of TYR enzyme
80%, and the dragon's blood extract that concentration is 800 μ g/mL can achieve 95% or more to the inhibiting rate of TYR enzyme.
Embodiment 3: dragon's blood extract and bamboo leaf flavone inhibit melanin production
Dragon's blood extract, bamboo leaf flavone and the two composition add appropriate DMSO to dissolve, and use cell culture
Base dilution obtains the medical fluid of 100 μ g/mL concentration.In different components, containing for Resina Draconis is 90%, 70%, 50%, 30%.
Cell culture: cell is inoculated in 1640 culture mediums containing 10% fetal calf serum, 37 DEG C, 5%CO after recovering2
It is cultivated in incubator, 0.25 pancreatin had digestive transfer culture.The B16 cell of logarithmic growth phase, after pancreatin digests at cell suspension after,
Add culture medium to dilute and be inoculated in 96 orifice plates, every empty 1000/100uL, culture is for 24 hours.Cell culture fluid is discarded, is added not of the same race
Normal incubation medium, every kind of 3 multiple holes are added in the pastille culture medium of class, blank control wells.The continuation of 96 orifice plates is incubated in the incubator
72h discards supernatant liquid, is cleaned cell 3 times using PBS.1mol/L sodium hydroxide solution, 80 DEG C of water-baths are added in each hole
1h measures each hole absorbance in the multi-functional continuous wavelength microplate reader of M1000 (Tecan company, Switzerland) at 490nm.Relative black
Plain production quantity=[(drug hole absorbance values-sodium hydroxide solution absorbance values)/(blank well absorbance values-
Sodium hydroxide solution absorbance values)].
Influence of 1 different pharmaceutical of table to melanin production
As it can be seen from table 1 dragon's blood extract and bamboo leaf flavone inhibit melanin to generate after combination
Effect becomes apparent compared with exclusive use.Bamboo leaf flavone does not have direct repression for tyrosinase, in the present invention
Melanin production is reduced by antioxidation with bamboo leaf flavone in composition and is conducive to that Resina Draconis is kept to extract
The stability of object, collaboration inhibit the generation of melanin.
Embodiment 4: dragon's blood extract Transdermal absorption
The preparation of Resina Draconis sample solution: dragon's blood extract 0.1g is dissolved in the ethanol solution of 10ml 30%, sufficiently
It is spare after dissolution.
The preparation of receiving liquid: drawing 15ml dehydrated alcohol and be placed in 50ml volumetric flask, and 0.9% physiological saline progress is added
It is spare as receiving liquid to prepare the normal saline solution for containing 30% ethyl alcohol for constant volume.
Mouse is taken off neck to put to death, takes a small amount of depilatory cream to be applied to abdomen and carries out depilation processing, skin physiological after clip depilation
Salt water is cleaned, and fascia is removed, and using Franz device, (acceptance pool 5.0ml receives pool area 1.13cm2), fixed device is connecing
By the physiological saline for (32 ± 0.5) DEG C that pre-temperature is added in room, clean mouse skin is fixed on the two of diffusion cell with clip
Between a half pond, stratum corneum side is subsequently placed in temperature (32 ± 0.5) DEG C water bath with thermostatic control, electromagnetic agitation speed to supply pool
(2000r/min).With Drug Percutaneous Absorption, receiving liquid side color is gradually deepened, and is in pale red.Respectively at 30,60,
90min takes 2ml receiving liquid and supplements 2ml with the receiving liquid of pre-temperature (32 ± 0.5) DEG C, keeps receiving liquid constancy of volume.
The HPLC of transdermal receiving liquid is detected
Waters HPLC PDA detector, chromatographic column KromasilC18 (250 × 4.6 ㎜), Detection wavelength: 275nm, column
Temperature: 40 DEG C of sampling volumes: 10 μ L, mobile phase: acetonitrile-water gradient, gradient are as follows:
Table 2
Dragon's blood extract and various time points receiving liquid high-efficient liquid phase chromatogram are as follows: dragon's blood extract efficient liquid phase
Chromatogram is as shown in Figure 2；30min Transdermal absorption ingredient high-efficient liquid phase chromatogram is as shown in Figure 3；60min Transdermal absorption ingredient is high
Effect liquid phase chromatogram figure is as shown in Figure 4；90min Transdermal absorption ingredient high-efficient liquid phase chromatogram is as shown in Figure 5.
From Fig. 2-Fig. 5 can be seen that liquid chromatography can detect in extract ingredient can Transdermal absorption, with when
Between elapse, each component content of Transdermal absorption is stepped up.
Embodiment 5: for inhibiting the preparation of the face cream of melanin
Based on face cream total weight, the weight composition of the face cream for inhibiting melanin is as follows: C12-C15Alkanol benzene
Formic acid esters 4%-6%, cyclopentasiloxane 4%-7%, cetanol 2.5%-4%, apricot kernel oil 2%-3%, Solsperse 2000 polyethers -2
2%-4%, -21 2%-4% of Solsperse 2000 polyethers, butanediol 2%-3%, EDTA-2Na 0.1%-0.3, glycerol 2%-4%,
Xanthan gum 0.2%-0.8%, carbomer 0.35%-0.45%, tocopherol acetate 1%-2%, hyaluronic acid 1%-2%, dragon
Sanguis Draxonis extract 0.25%-1%, bamboo leaf flavone 0.01%-0.11%, preservative 0.4%-0.8%, deionized water is extremely
100%.
Above-mentioned formula materials are weighed, Resina Draconis 1.00g, apricot kernel oil 4.00g are mixed, cetanol 5.00g, penta silicon of ring is added
Oxygen alkane 8.00g, C12-C15It is alkanol benzoic ether 12.00g, tocopherol palm ester 2.00g, -2 4.00g of Solsperse 2000 polyethers, hard
- 21 4.00g of resin acid alcohol polyethers, preservative (methyl hydroxybenzoate, ethyl hydroxy benzoate, Nipasol) 1.60g are as a phase；Weigh butanediol
4.00g, EDTA-2Na0.20g, glycerol 6.00g, xanthan gum 0.40g, carbomer 0.70g make after deionized water 135g mixing is added
For B phase；Niacinamide 2.00g, bamboo leaf flavone 0.11g and deionized water 10.00g, as c phase.A phase and b heat phase arrive
After 85 DEG C of dissolutions, after being cooled to 30 DEG C, c phase homogeneous is added in homogeneous, and vacuum outgas is canned to get the product for arriving Resina Draconis face cream.
Embodiment 6: for inhibiting the preparation of the face cream of melanin
Based on face cream total weight, the weight composition of the face cream for inhibiting melanin is as follows: C12-C15Alkanol benzene
Formic acid esters 4%-6%, cyclopentasiloxane 4%-7%, cetanol 2.5%-4%, apricot kernel oil 2%-3%, Solsperse 2000 polyethers -2
2%-4%, -21 2%-4% of Solsperse 2000 polyethers, butanediol 2%-3%, EDTA-2Na 0.1%-0.3, glycerol 2%-4%,
Xanthan gum 0.4%-0.8%, carbomer 0.35%-0.45%, tocopherol acetate 1%-2%, sodium ascorbyl phosphate 2%-
4%, hyaluronic acid 1%-2%, pawpaw sulfydryl enzyme 0.5%-1%, Bilberry fruit P.E 0.5%-1%, dragon's blood extract
0.25%-1%, bamboo leaf flavone 0.01%-0.11%, preservative 0.4%-0.8%, deionized water to 100%.
Above-mentioned formula materials are weighed, Resina Draconis 1.00g, apricot kernel oil 4.00g are mixed, cetanol 5.00g, penta silicon of ring is added
Oxygen alkane 8.00g, C12-C15Alkanol benzoic ether 12.00g, tocopherol palm ester 2.00g, Solsperse 2000 polyethers -24.00g, tristearin
Acid alcohol polyethers -214.00g, preservative (methyl hydroxybenzoate, ethyl hydroxy benzoate, Nipasol) 1.60g are as a phase；Weigh butanediol
4.00g, EDTA-2Na0.20g, glycerol 6.00g, xanthan gum 0.40g, carbomer 0.70g make after deionized water 129g mixing is added
For B phase；Weigh sodium ascorbyl phosphate 4.00g, niacinamide 2.00g, pawpaw sulfydryl enzyme 1.00g, Bilberry fruit P.E 1.00g, bamboo
Leaf flavone extract 0.11g and deionized water 10.00g, as c phase.After a phase and b heat phase to 85 DEG C of dissolutions, homogeneous is cooling
To after 30 DEG C, c phase homogeneous is added, vacuum outgas is canned to get the product for arriving Resina Draconis face cream.
The present inventor has carried out a large amount of detection to the face cream for being used to inhibit melanin in above-described embodiment 5 and 6 and has tested,
The result shows that being used to inhibit the face cream of melanin that can inhibit tyrosine activity in above-described embodiment 5 and 6, black is reduced
Element generates, and effectively prevents skin of face melanin deposition and color spot generates.
From above-described embodiment as can be seen that melanin inhibitor-dragon's blood extract provided by the present invention has well
Tyrosinase effect and percutaneous absorbability, can be as tyrosinase inhibitor for inhibiting melanin production；With regard to the present invention
For the provided external skin-care composition for inhibiting melanin, although bamboo leaf flavone itself is for tyrosinase
Without direct repression, but bamboo leaf flavone has preferable antioxidation to Resina Draconis, is conducive to dragon's blood
The stabilization exhausted.It can produce preferable synergistic effect after dragon's blood extract is compounded with bamboo leaf flavone, it is made
External skin-care composition is significantly greater than dragon's blood extract single dose to the inhibitory effect of melanin production；Using black of the invention
For inhibiting the face cream of melanin made of plain inhibitor or topical composition, tyrosine activity can be inhibited, reduce melanin
It generates, effectively prevents skin of face melanin deposition and color spot generates.
It should be noted that embodiment described above for explaining only the invention, is not constituted to of the invention any
Limitation.By referring to exemplary embodiments, invention has been described, it should be appreciated that word used in it is descriptive
With explanatory vocabulary, rather than limited vocabulary.The present invention can be made within the scope of the claims by regulation
Modification, and the present invention is revised in without departing substantially from scope and spirit of the present invention.Although the present invention described in it relates to
And specific method, material and embodiment, it is not intended that the present invention is limited to particular case disclosed in it, on the contrary, this hair
It is bright to can be extended to other all methods and applications with the same function.
Claims (11)
1. dragon's blood extract is preparing the application in the product for inhibiting melanin production, wherein the dragon's blood extract
The preparation method comprises the following steps: take 500g Resina Draconis crude drug, smash it through 80 meshes, the alcohol reflux of 8 times of volumes 60% added to extract 2 times,
It extracts 2 hours every time, filters merging filtrate, 55 DEG C of the condition in 55 DEG C of recycling ethyl alcohol on Rotary Evaporators, vacuum oven
Lower drying removes ethyl alcohol, obtains dragon's blood extract.
2. a kind of for inhibiting the external skin-care composition of melanin, which is characterized in that in the composition, Resina Draconis extracts
Total weight of the content of object based on composition is 90wt%-95wt%, and the composition also contains bamboo leaf flavone,
Wherein the dragon's blood extract the preparation method comprises the following steps: take 500g Resina Draconis crude drug, smash it through 80 meshes, add 8 times
The alcohol reflux of volume 60% extracts 2 times, extracts 2 hours every time, merging filtrate is filtered, in 55 DEG C of recycling second on Rotary Evaporators
Alcohol, it is dry under conditions of 55 DEG C in vacuum oven, ethyl alcohol is removed, dragon's blood extract is obtained.
3. it is a kind of for inhibiting the external-use skin care product of melanin, it is combined containing external-use skin care as described in claim 2
Object.
4. external-use skin care product according to claim 3, which is characterized in that in the external-use skin care product, based on outer
With the total weight of skin care, the dosage of the dragon's blood extract is 0.25wt%-1wt%.
5. external-use skin care product according to claim 3, which is characterized in that in the external-use skin care product, based on outer
With the total weight of skin care, the dosage of the external skin-care composition is 0.26wt-1.1wt%.
6. the external-use skin care product according to any one of claim 3-5, which is characterized in that the external-use skin care product
Exist by face cream, lotion, facial mask and in the form of at least one of washing baths.
7. a kind of for inhibiting the face cream of melanin, based on face cream total weight, the face cream includes:
Dragon's blood extract 0.25wt%-1wt%；
Bamboo leaf flavone < 1wt%；
C12-C15Alkanol benzoic ether 4wt%-6 wt%；
Cyclopentasiloxane 4wt%-7wt%；
Cetanol 2.5wt%-4wt%；
Apricot kernel oil 2wt%-3wt%；
- 2 2wt%-4wt% of Solsperse 2000 polyethers；
- 21 2wt%-4wt% of Solsperse 2000 polyethers；
Butanediol 2wt%-3wt%；
EDTA-2Na 0.1wt%-0.3wt%；
Glycerol 2wt%-4wt%；
Xanthan gum 0.2wt%-0.8wt%；
Carbomer 0.35wt%-0.45wt%；
Niacinamide 1wt%-2wt%；
Tocopherol palm ester 1wt%-2wt%；
Preservative 0.4wt%-0.8wt%；And
Water,
Wherein the dragon's blood extract the preparation method comprises the following steps: take 500g Resina Draconis crude drug, smash it through 80 meshes, add 8 times
The alcohol reflux of volume 60% extracts 2 times, extracts 2 hours every time, merging filtrate is filtered, in 55 DEG C of recycling second on Rotary Evaporators
Alcohol, it is dry under conditions of 55 DEG C in vacuum oven, ethyl alcohol is removed, dragon's blood extract is obtained.
8. face cream according to claim 7, which is characterized in that be based on face cream total weight, the bamboo leaf flavone
Dosage be 0.01wt%-0.11wt%.
9. face cream according to claim 7 or 8, which is characterized in that the face cream also includes pawpaw sulfydryl enzyme, cowberry extraction
One or more of object and sodium ascorbyl phosphate.
10. face cream according to claim 9, which is characterized in that be based on face cream total weight, the use of the pawpaw sulfydryl enzyme
Amount≤1wt%；Dosage≤1wt% of the Bilberry fruit P.E；Dosage≤4wt% of the sodium ascorbyl phosphate.
11. face cream according to claim 10, which is characterized in that it is based on face cream total weight, the pawpaw sulfydryl enzyme
Dosage is 0.5wt%-1wt%；The dosage of the Bilberry fruit P.E is 0.5wt%-1wt%；The dosage of the sodium ascorbyl phosphate
For 2wt%-4wt%.
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