Lightening compositions of multimachine reason restraint of tyrosinase activity and products thereof
Technical field
The present invention relates to a kind of lightening compositions, be specifically related to a kind of difference inhibition mechanism that generates rate-limiting enzyme-tryrosinase for melanin, the lightening compositions of Combinatorial Optimization comprising competitiveness, noncompetitive and linear hybrid competitive inhibition mechanism.
Background technology
The Asia women especially pays attention to pale U.S. of skin.For this demand, a large amount of cosmetics companies and researcher drop into manpower and materials and carry out the research and development of skin-whitening effective ingredient.Existing many skin-whitening agents come out at present, as early stage pure chemistry thing hydroquinone, the fruit acid in mid-term and tissue extract placenta cream and recent natural plant extracts such as arbutin, Vc derivant and Radix Glycyrrhizae extract etc.Consumers in general's demand has been satisfied in the appearance of these skin-whitening agents to a certain extent, has stimulated the development in skin-lightening cosmetic market.But we also should recognize clear-headedly, and the whitening effect of pure chemistry thing is better, but side effect is larger; Though the pure natural plant extract side effect is little, whitening effect is little or uncertain.
Present most skin-whitening agents is mainly for working in melanogenic rate-limiting enzyme-tryrosinase, existing research is known, and tyrosine → DOPA acid → DOPA quinone → dopachrome (or dihydroxy indole acid) → melanin oligomer → melanin is followed in the generation of dermal melanin.Can shift to epithelial cells by the melanocyte dendritic processes after melanin granule generates and arrive epidermis cell, thereby cause skin darkening.For the research of skin-whitening or spot-removing function, mainly for the inhibitory action of tryrosinase, as patent CN101080215A, CN101357114A, CN102614094A etc., have an effect as target spot for tyrosinase activity.
Although above research can suppress tyrosinase activity, but effect fails to reach expection in actual product, mainly there are several deficiencies: 1) fail further to study for the mechanism of restraint of tyrosinase activity, not yet study whitening agent or combination with which kind of mechanism restraint of tyrosinase activity; 2) known even if some whitening agent or composition suppress mechanism to tryrosinase, but not yet carry out the combination of working substance for various inhibition mechanism such as competitive inhibition, noncompetitive inhibition and linear noncompetitive inhibition mechanism; 3) fail to suppress mechanism and carry out the screening of whitening working substance or combination for difference, and make corresponding inhibition of enzyme activity test.Existing whitening agent or the combination that suppresses for tryrosinase, the often only competition inhibition effect of single consideration to tryrosinase, this action target spot is single, therefore has in actual use effect deficiency, required whitening agent dosage greatly and the deficiencies such as releasing stimulus.
Summary of the invention
The present invention is directed to the deficiency that existing tryrosinase suppresses mechanism research and technical scheme, comprehensively tyrosinase activity is suppressed different mechanism, select competitive tryrosinase inhibition, noncompetitive inhibition and the competitive natural tyrosinase inhibitor of linear hybrid, come screening design whitening working substance to make up; The effect of comprehensive three kinds of suppressor modes, thus improve whitening working substance and combination press down the enzyme effect, the biological effect that improves whitening also reduces the possible zest of using.
Technical scheme of the present invention is achieved in that
A kind of lightening compositions of multimachine reason restraint of tyrosinase activity, it contains the active matter of three kinds of restraint of tyrosinase active mechanisms: the active matter, the active matter of non-competitive inhibition and the active matter of linear hybrid competitive inhibition that play the effect of competition inhibition, wherein, the active matter that plays competition inhibition effect is at least a in ligustrazine, Morin, Radix Scutellariae extract; The active matter that plays non-competitive inhibition is at least a in hesperidin, Folium Ginkgo extract, Radix Rhodiolae extract; Playing the linear active matter that mixes competitive inhibition is Aloe cortex section's extract or paeonol.
Competition inhibition effect characteristics are: a competitive inhibitor is the analog of zymolyte often; The binding site of b inhibitor and enzyme is identical with the binding site of substrate and enzyme; The c inhibitor concentration is larger, and inhibitory action is larger; But increasing concentration of substrate can make the inhibition degree reduce; The d kinetic parameter: the Km value increases, and the Vm value is constant.
The non-competitive inhibition characteristics are: a inhibitor can be combined with the different parts of enzyme with substrate simultaneously; B must have substrate to exist, and inhibitor could produce inhibitory action to enzyme; C kinetic parameter Km value is constant, and Vm reduces.
As preferably, it forms and percentage by weight is: ligustrazine 20~40%; Morin 10~30%; Hesperidin 20~40%; Paeonol 30~40%.
As preferably, it forms and percentage by weight is: Morin 20~30%; Radix Scutellariae extract 10~30%; Radix Rhodiolae extract 20~40%; Aloe cortex section extract 20~30%.
As preferably, it forms and percentage by weight is: Morin 20~30%; Hesperidin 10~20%; Folium Ginkgo extract 20~40%; Aloe cortex section extract 20~30%.
A kind of cold cream, its formula and mass parts are as follows:
A component: join 12.0 parts of sugared amino stearic acid esters; 10.0 parts of spermol lactates; 5.0 parts, white liquid paraffin; 2.5 parts of the Polyethylene Glycol of cetostearyl alcohol; 1.0 parts of ceryl alcohols;
B component: 5.0 parts of propylene glycol; Spermol lactate 60 is 3.0 parts; 55.5 parts of distilled water;
C component: 0.5 part of Kazon; 0.5 part, essence
D component: 5.0 parts of lightening compositions.
A kind of preparation method of cold cream comprises the following steps: the A component is mixed, be heated to 80 ℃; Again the B component is mixed, be heated to 80 ℃; In stirring, the A component is added in the B component, adds lightening compositions and essence when temperature is down to 45 ℃, continue to be cooled to 25~28 ℃, packing.
A kind of cream pack, its formula and mass parts are as follows: 15.0 parts of polyvinyl acetate emulsions; 10.0 parts of polyvinyl alcohol; 3.0 parts, olive oil; 5.0 parts of glycerol; 8.0 parts of zinc oxide; 7.0 parts of Kaolin; 5.0 parts of ethanol; 40.0 parts of Purified Waters; 6.0 parts of lightening compositions; 0.5 part of spice; Kazon 0.5.
A kind of preparation method of cream pack comprises the following steps: with polyvinyl alcohol then and Kaolin with 3.0 parts of ethanol moistenings,, zinc oxide, Purified Water mixes, and is heated to 70 ℃ of stirrings, standing and be cooled to 45 ℃ after, with spice, antiseptic, olive oil and remaining ethanol mix, then and glycerol, lightening compositions, polyvinyl acetate emulsion add in the above-mentioned solution for preparing together, stir into pasty state and get final product.
A kind of clean skin cleansing milk, its formula and mass parts are as follows: Polyquaternium-10 is 1.0 parts; Cetearyl alcohol is 2.0 parts; Isopropyl palmitate is 6.0 parts; The cosmetic grade white oil is 0.5 part; Glycerol is 5.0 parts; Polysorbate-60 are 0.5 part; Glyceryl stearate is 0.5 part; Sodium lauryl sulfate is 0.3 part; Deionized water; 78.6 part; 5.0 parts of lightening compositions; 0.1 part of Kazon; 0.5 part, essence.
A kind of preparation method of clean skin cleansing milk is characterized in that comprising the following steps: with cetearyl alcohol, and isopropyl palmitate, the cosmetic grade white oil, glyceryl stearate, glycerol, polysorbate-60, sodium lauryl sulfate after deionized water mixes, adds Polyquaternium-10, heat to 85 ℃ of mixing and emulsifyings, be cooled to 50 ℃, add lightening compositions, Kazon, essence, stirring namely gets product.
A kind of fair complexion cream, its formula and mass parts are as follows:
A component: 6.0 parts, Cera Flava; 5.0 parts of hexadecanol; 8.0 parts of hydrogenated lanolins; 37.5 parts of squalanes; 4.0 parts of fatty acid glycerides;
B component: 2.0 parts of hydrophilic glyceryl monostearates; Polysorbate-20 are 2.0 parts; 0.25 part of cysteine; 5.0 parts of propylene glycol; 0.4 part of Kazon; 0.1 part of propyl gallate; 23.75 parts of distilled water;
C component: 1.0 parts, essence;
D component: 5.0 parts of lightening compositions.
A kind of manufacture method of fair complexion cream comprises the following steps: the A component is mixed, be heated to 75 ℃, under agitation, the A component is added the B component, lightening compositions and the essence, packing of adding during to 45 ℃ to be cooled.
The beneficial effect that the present invention compared with prior art has is: the difference for tyrosinase inhibitor suppresses machine-processed carry out screening and the combination of whitening working substance, use simultaneously three kinds to suppress the machine-processed novel composite inhibiting that draws, thereby that improves whitening working substance and combination presses down the enzyme effect, and the biological effect that improves whitening also reduces the possible zest of using; Broad covered area, the suitability are strong and good whitening and speckle dispelling effect arranged; And it can be designed to the products such as essence, day cream late frost, facial film, cleansing milk in cosmetic product.
The specific embodiment
Embodiment 1
Composition and the percentage by weight of the lightening compositions of the multimachine reason restraint of tyrosinase activity of the present embodiment are as follows:
Active matter or extract |
Proportioning (%) |
To the tryrosinase mechanism of action |
Ligustrazine |
20~40 |
The effect of competition inhibition |
Morin |
10~30 |
The effect of competition inhibition |
Hesperidin |
10~30 |
The noncompetitive inhibition effect |
Paeonol |
30~40 |
The effect of linear hybrid competition inhibition |
Embodiment 2
Composition and the percentage by weight of the lightening compositions of the multimachine reason restraint of tyrosinase activity of the present embodiment are as follows:
Active matter or extract |
Proportioning (%) |
To the tryrosinase mechanism of action |
Morin |
20~30 |
The effect of competition inhibition |
Radix Scutellariae extract |
10~30 |
The effect of competition inhibition |
Radix Rhodiolae extracting solution |
20~40 |
The noncompetitive inhibition effect |
Aloe cortex section extract |
20~30 |
The effect of linear hybrid competition inhibition |
Embodiment 3
Composition and the percentage by weight of the lightening compositions of the multimachine reason restraint of tyrosinase activity of the present embodiment are as follows:
Active matter or extract |
Proportioning (%) |
To the tryrosinase mechanism of action |
Morin |
20~30 |
The effect of competition inhibition |
Hesperidin |
10~20 |
The noncompetitive inhibition effect |
Folium Ginkgo extract |
20~40 |
The noncompetitive inhibition effect |
Aloe cortex section extract |
20~30 |
The effect of linear hybrid competition inhibition |
Embodiment 4
A kind of cold cream, its formula and mass parts are as follows:
A component: join 12.0 parts of sugared amino stearic acid esters; 10.0 parts of spermol lactates; 5.0 parts, white liquid paraffin; 2.5 parts of the Polyethylene Glycol of cetostearyl alcohol; 1.0 parts of ceryl alcohols;
B component: 5.0 parts of propylene glycol; Spermol lactate 60 is 3.0 parts; 55.5 parts of distilled water;
C component: 0.5 part of Kazon; 0.5 part, essence
5.0 parts of the lightening compositions of D component: embodiment 1.
[method for making] mixes the A component, is heated to 80 ℃; Again the B component is mixed, be heated to 80 ℃; In stirring, the A component is added in the B component, adds lightening compositions and the essence of embodiment 1 when temperature is down to 45 ℃, continue to be cooled to 25~28 ℃, packing.
[characteristic] this product pH value is 6.0, and near the acid-base value of application on human skin, therefore suitable people use, and can produce skin well to moisten and whitening effect.
Embodiment 5
A kind of cream pack, its formula and mass parts are as follows: 15.0 parts of polyvinyl acetate emulsions; 10.0 parts of polyvinyl alcohol; 3.0 parts, olive oil; 5.0 parts of glycerol; 8.0 parts of zinc oxide; 7.0 parts of Kaolin; 5.0 parts of ethanol; 40.0 parts of Purified Waters; 6.0 parts of the lightening compositions of embodiment 1; 0.5 part of spice; Kazon 0.5.
[method for making] with polyvinyl alcohol with 3.0 parts of ethanol moistenings, then and Kaolin, zinc oxide, Purified Water mixes, be heated to 70 ℃ of stirrings, standing and be cooled to 45 ℃ after, with spice, antiseptic, olive oil and remaining ethanol mix, again and glycerol, the lightening compositions of embodiment 1, polyvinyl acetate emulsion add in the above-mentioned solution for preparing together, stir into pasty state and get final product.
[characteristic] this cream pack has fine moisturizing function to skin of face after often using, and the effectively generation of check melanin, reaches the good result of whitening and speckle dispelling.
Embodiment 6:
A kind of clean skin cleansing milk, its formula and mass parts are as follows:
Polyquaternium-10 is 1.0 parts; Cetearyl alcohol is 2.0 parts; Isopropyl palmitate is 6.0 parts; The cosmetic grade white oil is 0.5 part; Glycerol is 5.0 parts; Polysorbate-60 are 0.5 part; Glyceryl stearate is 0.5 part; Sodium lauryl sulfate is 0.3 part; Deionized water; 78.6 part; 5.0 parts of the lightening compositions of embodiment 2; 0.1 part of Kazon; 0.5 part, essence.
A kind of preparation method of clean skin cleansing milk is characterized in that comprising the following steps: with cetearyl alcohol, and isopropyl palmitate, the cosmetic grade white oil, glyceryl stearate, glycerol, polysorbate-60, sodium lauryl sulfate after deionized water mixes, adds Polyquaternium-10, heat to 85 ℃ of mixing and emulsifyings, be cooled to 50 ℃, add lightening compositions, Kazon, the essence of embodiment 2, stirring namely gets product.
Embodiment 7
A kind of fair complexion cream, its formula and mass parts are as follows:
A component: 6.0 parts, Cera Flava; 5.0 parts of hexadecanol; 8.0 parts of hydrogenated lanolins; 37.5 parts of squalanes; 4.0 parts of fatty acid glycerides;
B component: 2.0 parts of hydrophilic glyceryl monostearates; Polysorbate-20 are 2.0 parts; 0.25 part of cysteine; 5.0 parts of propylene glycol; 0.4 part of Kazon; 0.1 part of propyl gallate; 23.75 parts of distilled water;
C component: 1.0 parts, essence;
5.0 parts of the lightening compositions of D component: embodiment 3.
[method for making] mixes the A component, is heated to 75 ℃, under agitation, the A component added the B component, lightening compositions and the essence that adds embodiment 3 during to 45 ℃ to be cooled, packing.
[characteristic] this frost has good moisture-keeping functions, and the effectively generation of check melanin in skin.
Above-mentioned composition is carried out the tryrosinase inhibition test of acellular system and cell system:
1, the inhibition test of lightening compositions to tryrosinase:
In the test system of 0.25mol/L phosphate buffer (pH6.8), respectively take the L-DOPA of L-Tyr, the 1.0mmol/L of 2.5mmol/L as substrate, the lightening compositions and the enzyme liquid that add respectively variable concentrations, the end reaction system is 1.0mL, enzyme concentration is 100U/mL, with the positive contrast of arbutin, deionized water is as the experiment blank; Measure optical density value D (λ) 475nm of 475nm during reaction 25min.Relative surplus vigor with enzyme is mapped to sample concentration, tries to achieve IC
50Value.
2, lightening compositions suppresses tryrosinase vitality test in the mouse melanin cell:
The B16 melanocyte is inoculated in 96 orifice plates, and every hole 100uL puts and adds medicine after in incubator, 24h is educated in cultivation.Take the positive contrast of arbutin, sterilization deionized water as blank.Abandoning supernatant after drug effect 3d, PBS with pH6.8 rinses twice, every hole adds 90uL and contains ultrasonication in the PBS ice bath that volume fraction is 1% Tritonx-100, every hole adds the L-Dopa of 10uL 10mmoL/L, hatch 60min for 37 ℃, in 475nm place colorimetric, will survey each hole absorbance after the blank well zeroing.
Suppression ratio=(1-medicine group mean light absorbency value ÷ matched group mean light absorbency value) * 100%.
3, finally suppress result:
According to suppressing result as can be known, no matter lightening compositions suppresses vigor at the tryrosinase of acellular system or cell line, and all the arbutin than single component is good.Especially show on the tryrosinase inhibition test to the B16 melanocyte, because having than single arbutin, the contained composition of said composition better sees through the cell performance, therefore the tryrosinase suppression efficiency of B16 melanocyte also had better performance, indicating that its effect to the human body whitening and speckle dispelling is better than the interpolation of one-component arbutin in various cosmetics add.
The above is only preferred embodiment of the present invention, and is in order to limit the present invention, within the spirit and principles in the present invention not all, any modification of doing, is equal to replacement, improvement etc., within all should being included in protection scope of the present invention.