WO2023103594A1 - 一种具有促进抗氧化酶表达功效的组合物及其应用 - Google Patents

一种具有促进抗氧化酶表达功效的组合物及其应用 Download PDF

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WO2023103594A1
WO2023103594A1 PCT/CN2022/125930 CN2022125930W WO2023103594A1 WO 2023103594 A1 WO2023103594 A1 WO 2023103594A1 CN 2022125930 W CN2022125930 W CN 2022125930W WO 2023103594 A1 WO2023103594 A1 WO 2023103594A1
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oil
expression
acid
promoting
composition
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English (en)
French (fr)
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叶睿
胡璠
胡晓芸
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懿奈(上海)生物科技有限公司
德肤科(上海)生物科技有限公司
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Publication of WO2023103594A1 publication Critical patent/WO2023103594A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—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
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9794—Liliopsida [monocotyledons]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/49—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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/08—Anti-ageing preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
    • A61K2800/59—Mixtures
    • A61K2800/592—Mixtures of compounds complementing their respective functions
    • 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

Definitions

  • the present application relates to the technical field of cosmetics, in particular to a composition with the effect of promoting the expression of antioxidant enzymes and its application.
  • Free radicals are very unstable, have strong oxidative properties, and have a strong ability to absorb electrons. They need to capture electrons from cell molecules in body tissues to pair themselves, and when cell molecules release an electron, they also become free radicals. , but also to snatch the electrons in the cell membrane or nucleus molecules, so that new free radicals are constantly generated. Free radicals can cause protein denaturation and cross-linking, causing many enzymes and hormones in the body to lose their biological activity, reducing the vitality of the body's immunity, nerve reflexes, and exercise capabilities. At the same time, it can also destroy the structure of nucleic acids and cause metabolic disorders in the entire body, etc. , and eventually cause disease in the body.
  • the purpose of this application is to provide a composition with the effect of promoting the expression of antioxidant enzymes and its application.
  • the first aspect of the present application provides a composition with the effect of promoting the expression of antioxidant enzymes, comprising turmeric root extract and dimethylmethoxychromanol, wherein the The mass ratio of the turmeric root extract to the dimethylmethoxychromanol is (0.5-3):1.
  • the ratio of the mass parts of the turmeric root extract to the dimethylmethoxychromanol is (1-2): 1.
  • the ratio of parts by mass of the turmeric root extract to the dimethylmethoxychromanol is 1:1 .
  • the second aspect of the present application provides the application of the composition having the effect of promoting the expression of antioxidant enzymes described in the first aspect of the present application in cosmetics and/or skin care products.
  • the cosmetic and/or skin care product is applied to the outer surface of the skin.
  • the cosmetic and/or skin care products are selected from one or more of eye cream, face cream, essence, lotion, and mask essence.
  • the composition having the effect of promoting the expression of antioxidant enzymes is dispersed in the matrix material as an additive, and the matrix material is selected from oily raw materials, powdery raw materials, and colloidal raw materials. one or several.
  • the cosmetics and/or skin care products also include: surfactants, flavors and fragrances, pigments, moisturizers, preservatives, antioxidants, ultraviolet absorbers, chelating agents, astringents One or more of additives, penetration aids, and nutritional additives.
  • the present application mixes turmeric root extract and dimethylmethoxychromanol according to a specific mass ratio, and achieves a synergistic effect in promoting the expression of antioxidant enzymes effect. Therefore, applying the composition provided by the present application with the effect of promoting the expression of antioxidant enzymes to cosmetics and/or skin care products can exert obvious anti-oxidation and anti-aging effects on the skin at a relatively low dosage.
  • Fig. 1 is the comparison chart of the impact of blank control group, model group, single administration group 1, single administration group 2, and experimental group 1 on the Hacat cell SOD activity induced by UVB;
  • Fig. 2 is the comparison chart of the impact of blank control group, model group, single administration group 1, single administration group 2, and experimental group 2 on the Hacat cell SOD activity induced by UVB;
  • Fig. 3 is the comparison chart of the impact of blank control group, model group, single administration group 1, single administration group 2, and experimental group 3 on the Hacat cell SOD activity induced by UVB;
  • Fig. 4 is the WB graph of the Hacat cell NQO1 protein expression induced by UVB by blank control group, model group, single administration group 1, single administration group 2, and experimental group 1;
  • Figure 5 is a comparison chart of the influence of blank control group, model group, single administration group 1, single administration group 2, and experimental group 1 on the expression of NQO1 protein in Hacat cells induced by UVB;
  • Fig. 6 is the WB graph of the Hacat cell NQO1 protein expression induced by UVB by blank control group, model group, single administration group 1, single administration group 2, and experimental group 2;
  • Figure 7 is a comparison chart of the influence of blank control group, model group, single administration group 1, single administration group 2, and experimental group 2 on the expression of NQO1 protein in Hacat cells induced by UVB;
  • Fig. 8 is a WB graph of UVB-induced Hacat cell NQO1 protein expression of blank control group, model group, single administration group 1, single administration group 2, and experimental group 3;
  • Figure 9 is a comparison chart of the effects of blank control group, model group, single administration group 1, single administration group 2, and experimental group 3 on the expression of NQO1 protein in Hacat cells induced by UVB.
  • An embodiment of the present application provides a composition with the effect of promoting the expression of antioxidant enzymes, comprising turmeric root extract and dimethylmethoxychromanol, wherein the turmeric root extract and the The ratio of parts by mass of dimethylmethoxychromanol is (0.5-3):1.
  • composition with the effect of promoting the expression of antioxidant enzymes comprising turmeric root extract and dimethylmethoxychromanol, wherein the turmeric root extract and the The ratio of parts by mass of dimethylmethoxychromanol is (1-2):1.
  • composition with the effect of promoting the expression of antioxidant enzymes comprising turmeric root extract and dimethylmethoxychromanol, wherein the turmeric root extract and the The ratio of parts by mass of dimethylmethoxychromanol is 1:1.
  • composition with the effect of promoting the expression of antioxidant enzymes provided by the embodiments of the present application can be applied to cosmetics and/or skin care products, and the cosmetics and/or skin care products are applied to the outer surface of the skin.
  • the cosmetic and/or skin care products using the composition having the effect of promoting the expression of antioxidant enzymes are selected from one or more of eye cream, face cream, essence, lotion, and mask essence.
  • the composition with the effect of promoting the expression of antioxidant enzymes is dispersed in the matrix material as an additive, and the matrix material is selected from oily raw materials, powdery raw materials, colloidal raw materials, oily raw materials, and solvent raw materials one or more of them.
  • the oleaginous material can be any one or several combinations of the following base materials: soybean oil, olive oil, almond oil, castor oil, peanut oil, cottonseed oil, tea seed oil, jojoba oil, avocado oil oil, coconut oil, palm oil, cocoa butter, rice bran oil, evening primrose oil, wheat germ oil, corn germ oil, palm wax, wood wax, mink oil, turtle oil, snake oil, egg butter, tallow, horse tallow, Pig fat, deer fat, lecithin, lanolin, beeswax, spermaceti, shellac wax, paraffin wax, petrolatum, microcrystalline wax, ozokerite, squalane, lanolin wax, lanolin alcohol, lanolin alcohol esters, Acetylated lanolin, acetylated lanolin alcohol, lanolin acid, lanolin esters, polyoxyethylene lanolin, polyoxyethylene lanolin ether, polyoxypropylene lanolin ether, hydrogenated lanolin, alkoxy
  • the Cn fatty acids such as: lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, erucic acid, capric acid.
  • the Cn fatty alcohols are such as: lauryl alcohol, cetyl alcohol, stearyl alcohol, isostearyl alcohol, oleyl alcohol, coconut oil alcohol.
  • the Cn fatty acid esters, Cn fatty alcohol esters such as: isopropyl myristate, myristyl myristate, myristyl lactate, cetyl lactate, isopropyl palmitate, palm tree isopropyl Capryl, Butyl Stearate, Isooctyl Stearate, Monoglyceryl Stearate, Polyethylene Glycol Stearate, Decyl Oleate, Glyceryl Caprylate, Glyceryl Caprate, Triolein ester.
  • the silty raw material can be any one or several combinations of the following matrix raw materials: mica, magnesium carbonate, titanium dioxide, zinc white powder, calcium hydroxide, calcium carbonate, kaolin, calcium stearate, zinc stearate , Magnesium Stearate, Talc, Silicon Dioxide, Aluminum Hydroxide, Calcium Pyrophosphate, Calcium Bicarbonate, Bentonite Clay.
  • the oily raw material can be any one or more of vegetable oily raw materials, animal oily raw materials, mineral oily raw materials, synthetic oily raw materials, and semi-synthetic oily raw materials.
  • vegetable oily raw materials animal oily raw materials, mineral oily raw materials, synthetic oily raw materials, and semi-synthetic oily raw materials.
  • the solvent raw material can be any one or several combinations of the following matrix raw materials: water, ethanol, propanol, isopropanol, isobutanol, ethylene glycol, 1,2-propylene glycol, cyclohexane , dichlorodifluoromethane, tetrafluorodichloroethane, ethylene glycol monoethyl ether, diethylene glycol monoethyl ether, acetone, methyl isobutyl ketone, ethyl acetate, butyl acetate, amyl acetate, phthalic acid Dibutyl ester.
  • matrix raw materials water, ethanol, propanol, isopropanol, isobutanol, ethylene glycol, 1,2-propylene glycol, cyclohexane , dichlorodifluoromethane, tetrafluorodichloroethane, ethylene glycol monoethyl ether, diethylene glycol
  • the cosmetics and/or skin care products using the composition with the effect of promoting the expression of antioxidant enzymes also include: surfactants, flavors and fragrances, pigments, moisturizers, preservatives, antioxidants , one or more of UV absorbers, chelating agents, astringents, penetration aids, pH regulators, and nutritional additives.
  • the surfactant can be: 1) anionic surfactant, such as: C12-14 fatty alcohol ammonium sulfate, C12-14 fatty alcohol polyoxyethylene ether carboxylate, lauryl sulfate diethanolamine salt, C12 -14 Fatty Alcohol Polyoxyethylene Ether Ammonium Sulfate, Triethanolamine Lauryl Sulfate, Alcohol Ether Phthalate Monoester Sodium Salt, Acylated Peptide, Sodium Lauric Acid Monoethanolamide Sulfate, n-Octyl Polyoxypropylene Ether Amber Acid monoester sulfonate, cetyl alcohol succinic acid monoester sulfonate, sulfosuccinic acid monoester disodium salt, succinic acid monostearyl disodium salt, alcohol ether sulfosuccinic acid monoester disodium salt, Fatty alcohol polyoxyethylene ether (sodium) sulfosuccinic acid monoester ammonium salt,
  • the essence and spices can be selected from: 1) natural spices, such as absinthe oil, eucalyptus oil, star anise oil, lime oil, white clove extract, clove oil, white orchid oil, white orchid extract , white orchid leaf oil, cypress oil, peppermint oil, atractylodes stearin, sweet rhinoceros extract, orange leaf oil, clove basil oil, big flower jasmine extract, wintergreen oil, angelica oil, angelica extract, lichenic acid, sandalwood oil , Toi Toi Flower Oil, Toi Toi Leaf Oil, Hyacinth Extract, Liquidambar Extract, Cinnamomum Oil, Sassafras Oil, Nardine Oil, Osmanthus Extract, Tangerine Oil, Carrot Seed Oil, Castoreum, Ambrette oil, yellow orchid oil, patchouli oil, artemisia oil, gourami extract, acacia extract, ginger oil, orange oil, peppermint oil, chrysanthemum extract, cumin oil, amber, am
  • the pigment can be one or more selected from the following components: zirconium dioxide, lead acetate, silver nitrate, chlorophyll, copper chlorophyllin, ferric ferrocyanide, guanine, fast red, food red I , Vat Red I, Carmine, Edible Bayberry Red, Edible Cherry Red, Food Red 17, Amaranth, Fast Green, Food Yellow 3, Edible Lemon Yellow, Edible Sudan Yellow, Edible Indigo, Food Blue 2, Fluorescent Pink, ⁇ -Carotene, Henna, Gardenia Red Pigment, Gardenia Green Pigment, Gardenia Blue Pigment, Gardenia Yellow Pigment, Titanium Mica Pearlescent Pigment, Iron Oxide Red, Iron Oxide Yellow, Iron Oxide Black, Chrome Oxide Green, Oxychlorine Bismuth, basic peach red, vermilion R, quinoline yellow, shikonin, lac red pigment, ultramarine blue, acid red 87, acid green 25, acid fluorescent yellow, acid orange 7, golden light red 53, Lisolbao Red
  • the preservative can be one or several combinations selected from the following components: sorbic acid and its salts, propionate, parabens, benzyl alcohol, benzoic acid and its salts, 5- Chloro-2-methyl-4-isothiazolin-3-one, -2-methyl-4-isothiazolin-3-one, carson, imidazolidinyl urea, dehydroacetic acid and its salts, sodium sulfite, Sodium bisulfite, sodium metabisulfite, 2,2-bromo-2-nitro-1,3-propanediol (Bropol), isoascorbic acid and its salts, kojic acid and its esters, azelaic acid, usnic acid , Resorcinol.
  • the antioxidant can be one or several combinations selected from the following components: SOD, butyl hydroxyanisole, gallate, ascorbic acid and its salts, erythorbic acid and its salts, sitosterol, reed Ding, tocopherol.
  • the moisturizing agent can be one or several combinations selected from the following components: 1,2-butanediol, sorbitol, xylitol, glycerin, lactate, polyethylene glycol, DL- Pyrrolidone carboxylate, D-glucose, kojic acid and its esters, uric acid, orotic acid, pectinic acid, laminin, collagen, trehalose, cholesteryl 12-hydroxystearate.
  • the ultraviolet absorber can be one or several combinations selected from the following components: pyridoxine hydrochloride, kojic acid and esters thereof, rutin, aloin, caffeine, hesperetin, 2-Ethylhexyl salicylate, 2-ethylhexyl p-dimethylaminobenzoate, 2-ethylhexyl p-methoxycinnamate, phenyl salicylate, anthranilate , 2-Hydroxy-4-methoxybenzophenone, 2-hydroxy-4-methoxybenzophenone-5-sulfonic acid, 2-hydroxy-4-octyloxybenzophenone.
  • the chelating agent may be one or several combinations selected from the following components: EDTA disodium salt, bipyridinethione zinc, and sodium thiosulfate.
  • the astringent can be one or several combinations selected from the following components: zinc phenol p-sulfonate, aluminum hydroxychloride, aluminum sulfate, potassium aluminum sulfate, zinc sulfate, aluminum chloride, zinc chloride , Calamine.
  • the penetration aid may be one or several combinations selected from the following components: dimethyl sulfoxide, 1-n-dodecylazepan-2-one, and 2-aminoethanol.
  • the pH regulator may be one or several combinations selected from the following components: tartaric acid, lactic acid, boric acid, and succinic acid.
  • the nutritional additive can be one or several combinations selected from the following components: chitin, chitosan, silk peptide, silk fibroin, lecithin, epidermal growth factor, trehalose, lactose, pearl Powder, collagen, collagen hydrolyzate, hyaluronic acid and its salts, egg membrane protein, phytosterol, lysozyme chloride, sodium chondroitin sulfate, bee milk, propolis, honey, glycyrrhizic acid, glycyrrhetinic acid, gamma-flax Acid, kojic acid and its esters, gibberellic acid, glycine, L-aspartic acid and its salts, L-cystine and its salts, DL-alanine, L-serine, DL-serine, L-methionine , DL-methionine, L-lysine hydrochloride, L-threonine, inos, in
  • the instruments, kits, cells and reagents used in the experimental examples can be purchased through open commercial channels, and are commonly used instruments, kits, cells and reagents in this field, and the operation steps of the instruments and kits are guided by the instructions conduct.
  • the SOD detection kit was used to detect the effect of cosmetic raw materials on the expression level of antioxidant enzymes, and the effect of cosmetic raw materials on the expression level of NQO1 protein was detected by Western Blot.
  • Superoxide dismutase Superoxide Dismutase, SOD
  • SOD can catalyze the dismutation of superoxide anion to generate hydrogen peroxide (H 2 O 2 ) and oxygen (O 2 ), which is an important antioxidant enzyme in organisms.
  • Quinone oxidoreductase 1 (NQO1) is a homodimeric flavinase that prevents oxidative damage to DNA by environmental stressors.
  • NQO1 plays an important role in protecting endogenous antioxidants by maintaining the reduced forms of ubiquinone and ⁇ -tocopheryl quinone.
  • Raw material 1 name: turmeric root extract; trade name: Tego Turmerone; properties: oily; purchased from Evonik Specialty Chemicals (Shanghai) Co., Ltd.
  • Raw material 2 name: dimethylmethoxychromanol; trade name: LIPOCHROMAN; properties: light brown to brown powder; purchased from Lubrizol Management (Shanghai) Co., Ltd.
  • Cell species Hacat human immortalized keratinocytes. Source: Cell Bank of Wuhan University, Type Culture Collection Committee, Chinese Academy of Sciences. Cell type: adherent cells.
  • test substance and the control substance are prepared into a mother solution with DMSO or ddH 2 O, and then diluted to an appropriate concentration with serum-free medium or PBS to form the following experimental groups:
  • Dosing composition (mass part ratio) Dosing concentration Blank control group ⁇ ⁇ model group ⁇ ⁇ Single administration group 1 lipochrome 20 ⁇ g/mL
  • Detection steps Take cells in the exponential growth phase to make a single cell suspension, inoculate on a 6-well plate at a density of 1.5 ⁇ 10 5 cells/mL, and incubate with a medium containing 10% fetal bovine serum at 37°C for 24 hours. The cells adhered to the wall, and the final volume of culture was 2 mL. Then the supernatant was discarded, and serum-free medium or freshly prepared serum-free medium containing different concentrations of test substances were added to incubate, and the final volume of culture was 2 mL.
  • the cells were irradiated with UVB (50J/cm 2 , 5min) using a UV lamp, and after culturing at 37°C for 16h, the SOD activity was detected using Beyond’s total SOD activity detection kit.
  • UVB 50J/cm 2 , 5min
  • Total Superoxide Dismutase Assay Kit with WST-8 (Total Superoxide Dismutase Assay Kit with WST-8) is a colorimetric reaction based on WST-8, which is used to detect cells, tissues or other samples by colorimetry
  • SOD is a kit for superoxide dismutase activity. The specific operation steps are carried out according to the instructions of the kit.
  • Fig. 1 is the comparison chart of the impact of blank control group, model group, single administration group 1, single administration group 2, and experimental group 1 on the Hacat cell SOD activity induced by UVB;
  • Fig. 2 is the comparison chart of the impact of blank control group, model group, single administration group 1, single administration group 2, and experimental group 2 on the Hacat cell SOD activity induced by UVB;
  • Figure 3 is a comparison chart of the effects of the blank control group, the model group, the single administration group 1, the single administration group 2, and the experimental group 3 on the SOD activity of Hacat cells induced by UVB.
  • Detection steps Take cells in the exponential growth phase to make a single cell suspension, inoculate on a 6-well plate at a density of 1.5 ⁇ 10 5 cells/mL, and incubate with a medium containing 10% fetal bovine serum at 37°C for 24 hours. The cells adhered to the wall, and the final volume of culture was 2 mL. Then the supernatant was discarded, and serum-free medium or freshly prepared serum-free medium containing different concentrations of test substances were added to incubate, and the final volume of culture was 2 mL.
  • the cells were irradiated with UVB (50J/cm 2 , 5min) using a UV lamp, and after culturing at 37°C for 16h, the protein content of NQO1 was detected by Western Blot.
  • UVB 50J/cm 2 , 5min
  • Extraction method of total protein add lysate into 6-well plate, lyse on ice for 30min, scrape off cells with cell scraper and transfer to centrifuge tube. Centrifuge at 4°C and 12000rpm for 5 minutes, take the supernatant and put it in a new centrifuge tube, add 5 ⁇ loading buffer according to the volume of the supernatant and boil in boiling water for 10 minutes to denature the protein, measure the protein concentration after cooling, and place in- Store at 80°C.
  • Fig. 4 is the NQO1 protein expression WB figure of blank control group, model group, single administration group 1, single administration group 2, experimental group 1 after administration of Hacat cells induced by UVB;
  • Figure 5 is a comparison chart of the influence of blank control group, model group, single administration group 1, single administration group 2, and experimental group 1 on the expression of NQO1 protein in Hacat cells induced by UVB;
  • Fig. 6 is the NQO1 protein expression WB figure of blank control group, model group, single administration group 1, single administration group 2, experimental group 2 after administration of Hacat cells induced by UVB;
  • Figure 7 is a comparison chart of the influence of blank control group, model group, single administration group 1, single administration group 2, and experimental group 2 on the expression of NQO1 protein in Hacat cells induced by UVB;
  • Fig. 8 is the NQO1 protein expression WB picture of blank control group, model group, single administration group 1, single administration group 2, experimental group 3 after administration of Hacat cells induced by UVB;
  • Figure 9 is a comparison chart of the effects of blank control group, model group, single administration group 1, single administration group 2, and experimental group 3 on the expression of NQO1 protein in Hacat cells induced by UVB.
  • n 3, Mean ⁇ SEM.
  • # means p ⁇ 0.05 compared with the control group
  • ## means p ⁇ 0.01 compared with the control group
  • * means p ⁇ 0.05 compared with the model group.

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Abstract

本申请涉及化妆品技术领域，公开了一种具有促进抗氧化酶表达功效的组合物及其应用。本申请提供的具有促进抗氧化酶表达功效的组合物，包含姜黄根提取物和二甲基甲氧基苯并二氢吡喃醇，其中，所述姜黄根提取物和所述二甲基甲氧基苯并二氢吡喃醇的质量份比值为(0.5～3)：1。本申请采用姜黄根提取物和二甲基甲氧基苯并二氢吡喃醇按照质量份比值进行混合，在促进抗氧化酶表达功效方面实现了协同增效作用。将本申请提供的具有促进抗氧化酶表达功效的组合物应用于化妆品和/或护肤品中，可在较低施用量下发挥明显的皮肤抗氧化、抗衰老作用。

Description

一种具有促进抗氧化酶表达功效的组合物及其应用
相关申请的交叉引用
本申请基于申请号为“202111509991.2”、申请日为2021年12月10日的中国专利申请提出，并要求该中国专利申请的优先权，该中国专利申请的全部内容在此以引入方式并入本申请。
技术领域
本申请涉及化妆品技术领域，特别涉及一种具有促进抗氧化酶表达功效的组合物及其应用。
背景技术
在机体代谢过程中，氧的一个电子被夺去后，不具有成对电子的氧原子就成为了自由基。自由基很不稳定，具有强氧化性，吸收的电子能力极强，它需要在身体组织的细胞分子中再夺取电子来使自己配对，而当细胞分子推陈出新一个电子后，它也变成自由基，又要去抢夺细胞膜或细胞核分子中的电子，这样不断产生新的自由基。自由基能引起蛋白质变性和交联，使体内的许多酶及激素失去生物活性，机体的免疫能力、神经反射能力、运动能力等系统活力降低，同时还能破坏核酸结构和导致整个机体代谢失常等，最终使机体发生病变。
从空气中吸入人体的氧气在酶的作用下，有相当一部分氧转化成超氧阴离子，用于消灭病毒等外来入侵者，起到保护机体的作用，而过剩的超氧阴离子则成为促使人体细胞衰老的自由基。人体自身可以产生内源性抗氧化物质催化分解超氧阴离子自由基，使人体达到安全平衡状态。但随着年龄的 增长，人体产生的内源性抗氧化水平会逐渐降低，此时过剩的超氧阴离子自由基就会攻击细胞膜上的不饱和磷脂，使细胞过早地老化，产生色素沉积，形成老年斑，导致人体衰老。
发明内容
本申请的目的在于提供一种具有促进抗氧化酶表达功效的组合物及其应用。
为解决上述技术问题，本申请的第一方面提供一种具有促进抗氧化酶表达功效的组合物，包含姜黄根提取物和二甲基甲氧基苯并二氢吡喃醇，其中，所述姜黄根提取物和所述二甲基甲氧基苯并二氢吡喃醇的质量份比值为(0.5～3)：1。
优选地，本申请提供的具有促进抗氧化酶表达功效的组合物中，姜黄根提取物和所述二甲基甲氧基苯并二氢吡喃醇的质量份比值为(1～2)：1。
更优选地，本申请提供的具有促进抗氧化酶表达功效的组合物中，所述姜黄根提取物和所述二甲基甲氧基苯并二氢吡喃醇的质量份比值为1：1。
本申请的第二方面提供了本申请第一方面所述的具有促进抗氧化酶表达功效的组合物在化妆品和/或护肤品中的应用。
本申请所提供的应用中，所述化妆品和/或护肤品施用于皮肤外表面。
优选地，本申请所提供的应用中，所述化妆品和/或护肤品选自眼霜、面霜、精华、乳液、面膜精华液中的一种或几种。
优选地，本申请所提供的应用中，所述具有促进抗氧化酶表达功效的组合物作为添加剂分散于基质原料中，所述基质原料选自油质原料、粉质原料、胶质原料中的一种或几种。
优选地，本申请所提供的应用中，所述化妆品和/或护肤品还包括：表 面活性剂、香精与香料、色素、保湿剂、防腐剂、抗氧剂、紫外线吸收剂、螯合剂、收敛剂、助渗剂、营养性添加剂中的一种或几种。
相对于现有技术而言，本申请采用姜黄根提取物和二甲基甲氧基苯并二氢吡喃醇按照特定质量份比值进行混合，在促进抗氧化酶表达功效方面实现了协同增效作用。因而，将本申请提供的具有促进抗氧化酶表达功效的组合物应用于化妆品和/或护肤品中，可在较低施用量的情况下发挥明显的皮肤抗氧化、抗衰老作用。
附图说明
图1为空白对照组、模型组、单独给药组1、单独给药组2、实验组1对UVB诱导的Hacat细胞SOD活力的影响对比图；
图2为空白对照组、模型组、单独给药组1、单独给药组2、实验组2对UVB诱导的Hacat细胞SOD活力的影响对比图；
图3为空白对照组、模型组、单独给药组1、单独给药组2、实验组3对UVB诱导的Hacat细胞SOD活力的影响对比图；
图4为空白对照组、模型组、单独给药组1、单独给药组2、实验组1对UVB诱导的Hacat细胞NQO1蛋白表达WB图；
图5为空白对照组、模型组、单独给药组1、单独给药组2、实验组1对UVB诱导的Hacat细胞NQO1蛋白表达影响对比图；
图6为空白对照组、模型组、单独给药组1、单独给药组2、实验组2对UVB诱导的Hacat细胞NQO1蛋白表达WB图；
图7为空白对照组、模型组、单独给药组1、单独给药组2、实验组2对UVB诱导的Hacat细胞NQO1蛋白表达影响对比图；
图8为空白对照组、模型组、单独给药组1、单独给药组2、实验组3 对UVB诱导的Hacat细胞NQO1蛋白表达WB图；
图9为空白对照组、模型组、单独给药组1、单独给药组2、实验组3对UVB诱导的Hacat细胞NQO1蛋白表达影响对比图。
具体实施方式
为使本申请的目的、技术方案和优点更加清楚，下面将对本申请的各实施方式进行详细的阐述。然而，本领域的普通技术人员可以理解，在本申请各实施方式中，为了使读者更好地理解本申请而提出了许多技术细节。但是，即使没有这些技术细节和基于以下各实施方式的种种变化和修改，也可以实现本申请所要求保护的技术方案。
本申请的实施方式提供一种具有促进抗氧化酶表达功效的组合物，包含姜黄根提取物和二甲基甲氧基苯并二氢吡喃醇，其中，所述姜黄根提取物和所述二甲基甲氧基苯并二氢吡喃醇的质量份比值为(0.5～3)：1。
在本申请部分实施方式提供一种具有促进抗氧化酶表达功效的组合物，包含姜黄根提取物和二甲基甲氧基苯并二氢吡喃醇，其中，所述姜黄根提取物和所述二甲基甲氧基苯并二氢吡喃醇的质量份比值为(1～2)：1。
在本申请部分实施方式提供一种具有促进抗氧化酶表达功效的组合物，包含姜黄根提取物和二甲基甲氧基苯并二氢吡喃醇，其中，所述姜黄根提取物和所述二甲基甲氧基苯并二氢吡喃醇的质量份比值为1：1。
本申请的实施方式所提供的具有促进抗氧化酶表达功效的组合物可应用于化妆品和/或护肤品中，所述化妆品和/或护肤品施用于皮肤外表面。
在本申请部分实施方式中，应用所述具有促进抗氧化酶表达功效的组合物的化妆品和/或护肤品选自眼霜、面霜、精华、乳液、面膜精华液中的一种或几种。
本申请部分实施方式中，所述具有促进抗氧化酶表达功效的组合物作为添加剂分散于基质原料中，所述基质原料选自油质原料、粉质原料、胶质原料、油脂原料、溶剂原料中的一种或几种。
其中，所述油质原料可以是下述基质原料中的任意一种或几种组合：大豆油、橄榄油、杏仁油、蓖麻油、花生油、棉籽油、茶籽油、霍霍巴油、鳄梨油、椰子油、棕榈油、可可脂、米糠油、月见草油、小麦胚芽油、玉米胚芽油、棕榈蜡、木蜡、水貂油、海龟油、蛇油、蛋黄油、牛脂、马脂、猪脂、鹿脂、卵磷脂、羊毛脂、蜂蜡、鲸蜡、虫胶蜡、石蜡、凡士林、微晶蜡、地蜡、角鲨烷、羊毛脂蜡、羊毛脂醇、羊毛脂醇酯化物、乙酰化羊毛脂、乙酰化羊毛脂醇、羊毛脂酸、羊毛脂酸酯化物、聚氧乙烯羊毛脂、聚氧乙烯羊毛醇醚、聚氧丙烯羊毛醇醚、氢化羊毛脂、烷氧基化氢化羊毛脂、硅油(如：二甲基硅油、八甲基硅油、甲基苯基聚硅氧烷)、聚硅氧烷-聚氧烷基嵌段共聚物、甲基含氢硅油、Cn脂肪酸及其酯、Cn脂肪醇及其酯；其中，n≥10，并优选为10-24，更优选为12-20，如14、16、17、18。
所述Cn脂肪酸如：月桂酸、肉豆蔻酸、棕榈酸、硬脂酸、油酸、亚油酸、芥酸、癸酸。所述Cn脂肪醇如：月桂醇、鲸蜡醇、硬脂醇、异硬脂醇、油醇、椰油醇。所述Cn脂肪酸酯、Cn脂肪醇酯如：肉豆蔻酸异丙酯、肉豆蔻酸肉豆蔻酯、十四醇乳酸酯、十六醇乳酸酯、棕榈酸异丙酯、棕榈树异辛酯、硬脂酸丁酯、硬脂酸异辛酯、硬脂酸单甘油脂、硬脂酸聚乙二醇酯、油酸癸酯、辛酸甘油酯、癸酸甘油酯、甘油三油酸酯。
所述粉质原料可以是下述基质原料中的任意一种或几种组合：云母、碳酸镁、钛白粉、锌白粉、氢氧化钙、碳酸钙、高岭土、硬脂酸钙、硬脂酸锌、硬脂酸镁、滑石粉、二氧化硅、氢氧化铝、焦磷酸钙、碳酸氢钙、膨润土。
其中，所述油脂原料可以是植物油质原料、动物油质原料、矿物油质 原料、合成油质原料、半合成油质原料中的任意一种或几种。如：淀粉、环糊精、黄原胶、角叉胶、瓜尔豆胶、阿拉伯胶、黄芪胶、琼脂、虫胶、海藻酸钠、明胶、甲基纤维素、乙基纤维素、羧甲基纤维素钠、羟乙基纤维素、羟丙基纤维素、聚纤维素醚季铵盐、聚乙烯醇、聚乙烯吡咯烷酮、聚丙烯酸钠、聚氧乙烯、乙烯基吡咯烷酮/乙酸乙烯酯共聚物、卡波树脂、硅酸铝镁、C12-15醇苯甲酸酯。
其中，所述溶剂原料可以是下述基质原料中的任意一种或几种组合：水、乙醇、丙醇、异丙醇、异丁醇、乙二醇、1,2-丙二醇、环己烷、二氯二氟甲烷、四氟二氯乙烷、乙二醇单乙醚、二甘醇单乙醚、丙酮、甲基异丁基甲酮、醋酸乙酯、醋酸丁酯、乙酸戊酯、邻苯二甲酸二丁酯。
在本申请部分实施方式中，应用所述具有促进抗氧化酶表达功效的组合物的化妆品和/或护肤品还包括：表面活性剂、香精与香料、色素、保湿剂、防腐剂、抗氧剂、紫外线吸收剂、螯合剂、收敛剂、助渗剂、pH值调节剂、营养性添加剂中的一种或几种。
其中，所述表面活性剂可以是：1)阴离子表面活性剂，如：C12-14脂肪醇硫酸铵、C12-14脂肪醇聚氧乙烯醚羧酸盐、十二烷基硫酸二乙醇胺盐、C12-14脂肪醇聚氧乙烯醚硫酸铵、十二烷基硫酸三乙醇胺、醇醚邻苯二甲酸单酯钠盐、酰化肽、月桂酸单乙醇酰胺硫酸钠、正辛醇聚氧丙烯醚琥珀酸单酯磺酸盐、十六醇琥珀酸单酯磺酸盐、磺基琥珀酸单酯二钠盐、琥珀酸单十八酯二钠盐、醇醚磺基琥珀酸单酯二钠盐、脂肪醇聚氧乙烯醚(钠)磺基琥珀酸单酯铵盐、油酰胺磺基琥珀酸单酯二钠盐、N-酰基谷氨酸钾盐、吡咯烷酮羧酸钠、聚丙烯酸钠、脂肪醇聚氧乙烯醚磷酸酯、脂肪醇聚氧乙烯醚磷酸单酯及其盐、脂肪醇聚氧乙烯醚磷酸单酯乙醇胺盐、烷基酚聚氧乙烯醚磷酸单酯及其盐、壬基酚醚磷酸单酯乙醇胺盐、烷基磷酸酯盐、邻苯二甲酸单月桂醇酯盐、十一碳烯酸锌、阴离子型氨基酸表面活性剂、N-C12-18酰基谷氨酸钠、N-月桂酰L-丙氨酸钠、N-硬脂酰基谷氨酸单钠盐、N-椰子酰基谷 氨酸单钠盐、N-混合脂肪酰基谷氨酸单钠盐、长直链烷基芳醚磺酸钠TH；2)非离子表面活性剂，如：油醇聚氧乙烯醚(如乳化剂VO系列)、烷基酚聚氧乙烯醚(如乳化剂OPE系列)、硬脂酸聚氧乙烯酯、乙二醇单硬脂酸酯、二硬脂酸乙二醇酯、聚乙二醇单硬脂酸酯、聚乙二醇双硬脂酸酯、聚乙二醇双月桂酸酯、双硬脂酸甘油酯、三硬脂酸甘油酯、蔗糖硬脂酸酯、聚氧乙烯甘油醚单硬脂酸酯、N-月桂酰基谷氨酸双烷醇酯、苯甲酸脂肪醇酯、山梨醇单脂肪酸酯(司盘，Span，如司盘-20、司盘60、司盘65、司盘-80、司盘-83、司盘-85)、聚氧乙烯醚山梨醇单脂肪酸酯(吐温，Tween，如吐温-20、吐温-40、吐温-60、吐温-61、吐温-80)、二乙醇胺、三乙醇胺、脂肪酸单乙醇酰胺(如椰子油酸单乙醇酰胺、月桂酸单乙醇酰胺、棕榈酸单乙醇酰胺)、椰子油酸二乙醇酰胺、月桂酸二乙醇酰胺、甲基葡萄糖苷硬脂酸酯、聚氧乙烯甲基葡萄糖苷硬脂酸酯、乙二醇葡萄糖苷硬脂酸酯、丙二醇基葡萄糖苷硬脂酸酯、C8-16烷基葡萄糖苷、甘油葡萄糖苷硬脂酸酯、琥珀酸月桂醇醚单酯、月桂醇甘油酯聚氧乙烯醚、硬脂酸甘露醇酐酯、聚氧乙烯貂油、貂油酸甲酯、貂油酸异丙酯；3)阳离子表面活性剂，如氯化C12-18烷基三烷基铵(如氯化十六烷基二甲基烷基铵、氯化十八烷基三甲基铵、氯化十六烷基三甲基铵)、氯化油酰胺丙基二羟丙基二甲基铵、氯化二羟丙基二甲基十二烷基铵、氯化二甲基二烯丙基铵/丙烯酰胺共聚物、氯化油酰胺丙基-2,3-二羟丙基二甲基铵、乙基硫酸油酰胺丙基二甲基乙基铵、氯化双C12-18基二甲基铵、聚季铵-11调理剂、M-505聚季铵-7调理剂、貂油酰胺丙基胺-壳聚糖、貂油酰胺丙基胺-水解蛋白、阳离子蛋白肽、瓜尔胶-羟丙基三甲基氯化铵盐、聚纤维素醚季铵盐、单油酸三乙醇胺酯、DNP系列；4)两性离子表面活性剂，如C12-18烷基二羟乙基甜菜碱、N-C12-18烷基-N-(2-羟乙基)-N-(2-甲酰胺基乙基)铵乙酸盐、C12-18烷氧基羟丙基甜菜碱、氧化椰油酰胺基丙基胺、椰油羟乙基磺酸钠、氧化十八烷基二甲基胺、椰油酰二乙醇胺氧化胺、N-烷基-β-氨基丙酰二乙醇胺、氧化月桂酰基丙胺、N-C12-18酰基谷氨酸、羟乙基 癸酸咪唑啉甜菜碱、羟乙基肉豆蔻酸咪唑啉甜菜碱、羟乙基棕榈酸咪唑啉甜菜碱。
其中，所述香精与香料可以是选自：1)天然香料，如苦艾油、桉叶油、八角茴香油、白柠檬油、白丁香浸膏、丁香油、白兰花油、白兰花浸膏、白兰叶油、柏木油、薄荷油、苍术硬脂、草木犀浸膏、橙叶油、丁香罗勒油、大花茉莉浸膏、冬青油、当归油、当归浸膏、地衣酸、地檀香油、玳玳花油、玳玳叶油、风信子浸膏、枫香浸膏、芳樟油、黄樟油、甘松油、桂花浸膏、红橘油、胡萝卜籽油、海狸香、黄葵油、黄兰油、藿香油、黄蒿油、九里香浸膏、金合欢浸膏、姜油、橘子油、椒样薄荷油、菊花浸膏、枯茗油、龙涎香、留兰香油、灵猫香、腊梅油、铃兰浸膏、木香根油、米兰花油、玫瑰香叶油、玫瑰油、茉莉浸膏、牡蛎油、墨红浸膏、墨红油、柠檬桉叶油、柠檬油、七里香浸膏、楠叶油、芹菜籽油、肉豆蔻油、肉桂油、山苍子油、水貂香膏酊、水仙浸膏、松针油、树苔浸膏、素馨浸膏、麝香、麝鼠香膏、甜橙油、檀香油、晚香玉油、晚香玉浸膏、万寿菊精油、小茴香油、血桧木油、香茅油、香叶油、康乃馨油、紫苏油、香紫苏油、香附油、香柠檬薄荷油、岩兰草油、香薷油、香薷草浸膏、悬钩子蔷薇花油、悬钩子蔷薇花浸膏、薰衣草油、月桂叶油、依兰依兰油、芫荽籽油、鱼香草油、岩蔷薇浸膏、岩桂油、柚皮油、鸢尾油、烟草花油、香橼叶油、栀子花浸膏、紫穗槐油、紫罗兰叶浸膏、愈创木酚、4-甲基愈创木酚、4-乙基愈创木酚、天然香兰素、樟脑；2)合成香精，如柠檬烯、长叶烯、乙位石竹烯、异长叶烯、异长叶烯酮、异长叶烷酮、溴代苏合香烯、二苯甲烷、二苯醚、间甲基二苯醚、丁香酚、异丁香酚、乙位萘甲醚、乙位萘乙醚、对甲基茴香醚、异丁香酚苄醚、玫瑰醚、檀香醚、水仙醚(或茉莉醚)、降龙涎香醚、甲基柏木醚、环氧柏木烷、埃林塔尔、埃林塔尔N、3-己烯醇、癸醇、月桂醇、苯甲醇、苯乙醇、对甲氧基苄醇、肉桂醇、二甲基苄基甲醇、香叶醇、橙花醇、芳樟醇、玫瑰醇、松油醇、香茅醇、消旋薄荷脑、薄荷脑、冰片、柏木醇、柏木烷酮、甲基柏 木酮、3,4-二氧亚甲基苯甲醇、α-甲基-3,4-二氢亚甲基苯丙醛、2-叔丁基-4-甲基环己醇、香紫苏醇、檀香醇、3-甲基-5-(2’,2’,3’-三甲基环戊烯-1’-基)戊-2-醇、苯甲醛、苯乙酮、苯乙酸、对甲基苯乙酮二苯甲酮、月桂醛、肉桂醛、肉桂酸、α-戊基-β-苯基丙烯醛、α-苯基丙醛、铃兰醛、新铃兰醛、兔耳草醛、3,4-二氧亚甲基苯甲醛、3,7-二甲基-6-辛烯醛、3,7-二甲基-7-羟基辛醛、柠檬醛、乙位环高柠檬醛、艾薇醛、柑青醛、4-羟基-3-甲氧基苯甲醛、α-二氢突厥酮、β-突厥烯酮、紫罗兰酮、6-甲基紫罗兰酮、橙花酮、香芹酮、胡椒基丙酮、覆盆子酮、药草酮、顺式茉莉酮、庚醛乙二醇缩醛、苯乙醛二甲缩醛、辛醛二甲缩醛、α-戊基桂醛二甲缩醛、柠檬醛二甲缩醛、柠檬醛二乙缩醛、茴香醛二甲缩醛、蘑菇醛、叶青素、风信子素、苹果酯、吉普缩酮、甲酸香叶酯、甲酸香茅酯、阿弗曼酯、乙酸叶醇酯、乙酸苄酯、乙酸苯乙酯、乙酸肉桂酯、乙酸松油酯、乙酸龙脑酯、乙酸芳樟酯、乙酸香叶酯、乙酸对叔丁基环己酯、乙酸邻叔丁基环己酯、乙酸香根酯、乙酸琥珀酯、乙酸阿弗曼酯、乙酸十氢萘酯、乙酸柏木酯、丙酸香叶酯、乙酸三氯甲基苯甲酯、丁酸苄酯、丁酸叶醇酯、丁酸香叶酯、2-甲基戊酸苯乙酯、异丁酸对甲酚酯、2-壬烯酸甲酯、苯甲酸乙酯、苯甲酸甲酯、苯甲酸叶醇酯、苯甲酸苄酯、苯乙酸乙酯、苯乙酸对甲酚酯、肉桂酸乙酯、水杨酸丁酯、水杨酸异丁酯、水杨酸异戊酯、水杨酸叶醇酯、水杨酸己酯、水杨酸苄酯、水杨酸苯乙酯、水杨酸环己酯、茉莉素、茉莉酯、2-戊基环戊酮醋酸甲酯、橙花素、γ-壬内酯、γ-十一内酯、香豆素、吲哚、3-甲基吲哚、3,7-二甲基-2,6-辛二烯腈、对甲氧基苯甲腈、二甲苯麝香、酮麝香、2,6-二硝基-3-甲氧基-4-叔丁基甲苯、1,1-二甲基-4-乙酰基-6-叔丁基茚满、六甲基三环异色满麝香、1,1,2,3,3,6-六甲基-5-乙酰基茚满、麝香-T、麝香-M、麝香-L、麝香-F、11-氧杂环十六内酯、吐纳麝香；3)香精，玫瑰香精、茉莉香精、檀香香精、白兰香精。
其中，所述色素可以是选自如下组分中的一种或几种：二氧化锆、醋酸铅、硝酸银、叶绿素、叶绿素铜、亚铁氰化铁、鸟嘌呤、坚牢红、食品红 I、还原红I、胭脂红、食用杨梅红、食用樱桃红、食品红17、苋菜红、坚牢绿、食品黄3、食用柠檬黄、食用苏丹黄、食用靛蓝、食品蓝2、荧光桃红、β-胡萝卜素、指甲花、栀子红色素、栀子绿色素、栀子蓝色素、栀子黄色素、钛云母珠光颜料、氧化铁红、氧化铁黄、氧化铁黑、氧化铬绿、氧氯化铋、碱性桃红、银朱R、喹啉黄、紫草宁、紫胶红色素、群青、酸性红87、酸性绿25、酸性荧光黄、酸性橙7、金光红53、立索尔宝红BK、立索尔宝红2R、永固橙。
其中，所述防腐剂可以是选自如下组分中的一种或几种组合：山梨酸及其盐、丙酸盐、对羟基苯甲酸酯、苯甲醇、苯甲酸及其盐、5-氯-2-甲基-4-异噻唑啉-3-酮、-2-甲基-4-异噻唑啉-3-酮、卡松、咪唑烷基脲、脱氢醋酸及其盐、亚硫酸钠、亚硫酸氢钠、焦亚硫酸钠、2,2-溴-2-硝基-1,3-丙二醇(布罗波尔)、异抗坏血酸及其盐、曲酸及其酯、壬二酸、松萝酸、间苯二酚。
其中，所述抗氧剂可以是选自如下组分中的一种或几种组合：SOD、丁基羟基茴香醚、没食子酸酯、抗坏血酸及其盐、异抗坏血酸及其盐、谷甾醇、芦丁、生育酚。
其中，所述保湿剂可以是选自如下组分中的一种或几种组合：1,2-丁二醇、山梨醇、木糖醇、甘油、乳酸盐、聚乙二醇、DL-吡咯烷酮羧酸盐、D-葡萄糖、曲酸及其酯、尿酸、乳清酸、果胶酸、昆布氨酸、胶原蛋白、海藻糖、12-羟基硬脂酸胆甾醇酯。
其中，所述紫外线吸收剂可以是选自如下组分中的一种或几种组合：吡哆醇盐酸盐、曲酸及其酯、芦丁、芦荟苷、咖啡氨酸、橙皮素、2-乙基己基水杨酸酯、2-乙基己基对二甲基氨基苯酸酯、2-乙基己基对甲氧基肉桂酸酯、水杨酸苯酯、基邻氨基苯甲酸酯、2-羟基-4-甲氧基二苯甲酮、2-羟基-4-甲氧基二苯甲酮-5-磺酸、2-羟基-4-辛氧基二苯甲酮。
其中，所述螯合剂可以是选自如下组分中的一种或几种组合：EDTA 二钠盐、双吡啶硫酮锌、硫代硫酸钠。
其中，所述收敛剂可以是选自如下组分中的一种或几种组合：苯酚对磺酸锌、羟基氯化铝、硫酸铝、硫酸铝钾、硫酸锌、氯化铝、氯化锌、卡拉明。
其中，所述助渗剂可以是选自如下组分中的一种或几种组合：二甲基亚砜、1-正十二烷基氮杂环庚-2-酮、2-氨基乙醇。
其中，所述pH值调节剂可以是选自如下组分中的一种或几种组合：酒石酸、乳酸、硼酸、琥珀酸。
其中，所述营养性添加剂可以是选自如下组分中的一种或几种组合：甲壳素、壳聚糖、丝肽、丝素、卵磷脂、表皮细胞生长因子、海藻糖、乳糖、珍珠粉、胶原蛋白、胶原水解物、透明质酸及其盐、蛋膜素、植物甾醇、氯化溶菌酶、硫酸软骨素钠、蜂乳、蜂胶、蜂蜜、甘草酸、甘草次酸、γ-亚麻酸、曲酸及其酯、赤霉酸、甘氨酸、L-天冬氨酸及其盐、L-胱氨酸及其盐、DL-丙氨酸、L-丝氨酸、DL-丝氨酸、L-蛋氨酸、DL-蛋氨酸、L赖氨酸盐酸盐、L苏氨酸、肌醇、麦芽醇、泛酸盐、D-泛醇、视黄醇醋酸酯、尿囊素、吡哆醇盐酸盐、可的松、烟酰胺、维生素、雌激素、苏木素、尿酸、乳清酸、芦荟苷、壬二酸、松香酸、松萝酸、昆布氨酸、胸腺素、茜草素、柠檬烯、黄芩黄素、脱氧胆酸、愈疮木萸、橙皮素、鞣酸、二硫化硒、茶皂苷、柠檬酸三乙酯、丁基羟基甲苯、植酸、聚氯化二甲基二烯丙基铵。
实验例
实验例中用到的仪器、试剂盒、细胞和试剂均可通过公开的商业渠道购买，为本领域常用的仪器、试剂盒、细胞和试剂，仪器和试剂盒的使用操作步骤按照其说明书的指导进行。
实验目的：通过建立UVB诱导Hacat细胞的氧化损伤模型，使用SOD检测试剂盒检测化妆品原料对抗氧化酶表达水平的影响，Western Blot检测 化妆品原料对NQO1蛋白表达水平的影响。超氧化物歧化酶(Superoxide Dismutase,SOD)能催化超氧化物阴离子发生歧化作用，生成过氧化氢(H 2O 2)和氧气(O 2)，是生物体内一种重要的抗氧化酶。醌氧化还原酶1(NQO1)是一种同源二聚体黄素酶，NQO1能够阻止环境胁迫剂对DNA造成氧化损伤。此外，NQO1通过维持泛醌和α-生育酚醌的还原形式，在保护内源性抗氧化剂中起重要作用。
实验原料
原料1：名称：姜黄根提取物；商品名：Tego Turmerone；性状：油状；购自赢创特种化学(上海)有限公司。
原料2：名称：二甲基甲氧基苯并二氢吡喃醇；商品名：LIPOCHROMAN；性状：浅棕色至棕色粉末；购自路博润管理(上海)有限公司。
主要仪器与试剂
表1.实验所用主要仪器
Figure PCTCN2022125930-appb-000001
Figure PCTCN2022125930-appb-000002
表2.实验所用主要试剂
Figure PCTCN2022125930-appb-000003
细胞培养
细胞品种：Hacat人永生化角质细胞。来源：中国科学院典型培养物保藏委员会武汉大学细胞库。细胞类型：贴壁细胞。
培养条件：5％CO 2，37℃恒温培养箱。培养基：89％MEM+10％FBS+1％双抗。
受试物和对照品的配制方法：受试物和对照品用DMSO或者ddH 2O配制成母液，再用无血清培养基或者PBS稀释至合适浓度，形成如下实验分组：
表3实验分组
组别 给药组成(质量份比) 给药浓度
空白对照组 \ \
模型组 \ \
单独给药组1 lipochroman 20μg/mL
单独给药组2 姜黄根提取物 20μg/mL
实验组1 姜黄根提取物：lipochroman＝1:1 20μg/mL
实验组2 姜黄根提取物：lipochroman＝2:1 20μg/mL
实验组3 姜黄根提取物：lipochroman＝1:5 20μg/mL
试验方法
(一)抗氧化酶检测
1、检测步骤：取指数生长期的细胞制成单细胞悬液，以1.5×10 5cells/mL的密度接种于6孔板上，用含10％胎牛血清的培养基37℃培养24h使细胞贴壁，培养终体积为2mL。而后吸弃上清，加入无血清培养基或新鲜制备的含不同浓度受试物的无血清培养基孵育，培养终体积为2mL。培养12h/16h/24h/32h/48h/72h后，使用紫外灯对细胞进行UVB照射(50J/cm 2，5min)，37℃培养16h后使用碧云天的总SOD活性检测试剂盒检测SOD活性。
碧云天的总SOD活性检测试剂盒(WST-8法)(Total Superoxide Dismutase Assay Kit with WST-8)是一种基于WST-8的显色反应，通过比色来检测细胞、组织或其它样品中SOD即超氧化物歧化酶活性的试剂盒。具体操作步骤参照试剂盒的说明书进行。
2、数据录入和统计分析：采用Excel录入和Prism(Version7)软件包统计处理，以均数±标准误表示。各组间做单因素方差分析,认为p<0.05有统计学差异。
3、实验结果
图1为空白对照组、模型组、单独给药组1、单独给药组2、实验组1对UVB诱导的Hacat细胞SOD活力的影响对比图；
图2为空白对照组、模型组、单独给药组1、单独给药组2、实验组2 对UVB诱导的Hacat细胞SOD活力的影响对比图；
图3为空白对照组、模型组、单独给药组1、单独给药组2、实验组3对UVB诱导的Hacat细胞SOD活力的影响对比图。
图1中：n＝3，Mean±SEM。“###”表示与对照组相比p＜0.001，“*”表示与模型组相比p＜0.05，“**”表示与模型组相比p＜0.01，“***”表示与模型组相比p＜0.001。
根据图1，采用实验组1给药(按质量份计，姜黄根提取物：lipochroman＝1:1)，与单独给药组1(lipochroman单独给药)、单独给药组2(姜黄根提取物单独给药)相比，实验组1可显著增加Hacat细胞的SOD，说明实验组在促抗氧化酶表达方面具有协同增效作用。
根据图2，采用实验组2给药(按质量份计，姜黄根提取物：lipochroman＝2:1)，与单独给药组1(lipochroman单独给药)、单独给药组2(姜黄根提取物单独给药)相比，实验组2也可显著增加Hacat细胞的SOD活性。
根据图3，采用实验组3给药(按质量份计，姜黄根提取物：lipochroman＝1:5)，与单独给药组1(lipochroman单独给药)、单独给药组2(姜黄根提取物单独给药)相比，实验组3在增加Hacat细胞的SOD活性方面无明显效果。
(二)NQO1蛋白表达水平检测
1、检测步骤：取指数生长期的细胞制成单细胞悬液，以1.5×10 5cells/mL的密度接种于6孔板上，用含10％胎牛血清的培养基37℃培养24h使细胞贴壁，培养终体积为2mL。而后吸弃上清，加入无血清培养基或新鲜制备的含不同浓度受试物的无血清培养基孵育，培养终体积为2mL。培养12h/16h/24h/32h/48h/72h后，使用紫外灯对细胞进行UVB照射(50J/cm 2，5min)，37℃培养16h后使用Western Blot检测NQO1蛋白含量。
提取总蛋白方法：6孔板内加入裂解液，冰上裂解30min，用细胞刮 刮下细胞并转至离心管中。4℃，12000rpm离心5min，取上清液置于新的离心管中，根据上清液体积加入5×loading buffer后置于沸水中煮10min，进行蛋白变性，冷却后测定蛋白浓度，置于-80℃保存。
BCA法测定蛋白浓度：操作步骤按试剂盒说明书。(BCA试剂盒品牌：Biosharp)
Western blot(NQO1抗体，CST)：蛋白上样进行SDS-PAGE凝胶电泳，电泳后将蛋白条带转印至NC膜上，5％脱脂乳封闭后4℃孵育一抗过夜，洗膜后孵育二抗1h，最终使用ECL底物发光液显影条带。
2、数据录入和统计分析：采用Excel录入和Prism(Version7)软件包统计处理，以均数±标准误表示。各组间做单因素方差分析,认为p<0.05有统计学差异。
3、实验结果
图4为空白对照组、模型组、单独给药组1、单独给药组2、实验组1对UVB诱导的Hacat细胞给药后的NQO1蛋白表达WB图；
图5为空白对照组、模型组、单独给药组1、单独给药组2、实验组1对UVB诱导的Hacat细胞NQO1蛋白表达影响对比图；
图6为空白对照组、模型组、单独给药组1、单独给药组2、实验组2对UVB诱导的Hacat细胞给药后的NQO1蛋白表达WB图；
图7为空白对照组、模型组、单独给药组1、单独给药组2、实验组2对UVB诱导的Hacat细胞NQO1蛋白表达影响对比图；
图8为空白对照组、模型组、单独给药组1、单独给药组2、实验组3对UVB诱导的Hacat细胞给药后的NQO1蛋白表达WB图；
图9为空白对照组、模型组、单独给药组1、单独给药组2、实验组3对UVB诱导的Hacat细胞NQO1蛋白表达影响对比图。
其中，n＝3，Mean±SEM。“#”表示与对照组相比p＜0.05，“##”表示与对照组相比p＜0.01，“*”表示与模型组相比p＜0.05。
根据图4和图5，采用实验组1给药(按质量份计，姜黄根提取物：lipochroman＝1:1)，与单独给药组1(lipochroman单独给药)、单独给药组2(姜黄根提取物单独给药)相比，实验组1可显著提高Hacat细胞的NQO1蛋白表达水平。
根据图6和图7，采用实验组2给药(按质量份计，姜黄根提取物：lipochroman＝1:5)，与单独给药组1(lipochroman单独给药)、单独给药组2(姜黄根提取物单独给药)相比，实验组2在提高Hacat细胞的NQO1蛋白表达水平方面无明显效果。
根据图8和图9，采用实验组3给药(按质量份计：Et-VC：VC-IP：VE：NAC＝2:3:0.5:0.01)，与单独给药组1(lipochroman单独给药)、单独给药组2(姜黄根提取物单独给药)相比，实验组3在提高Hacat细胞的NQO1蛋白表达水平方面无明显效果。
本领域的普通技术人员可以理解，上述各实施方式是实现本申请的具体实施例，而在实际应用中，可以在形式上和细节上对其作各种改变，而不偏离本申请的精神和范围。

Claims (8)

  1. 一种具有促进抗氧化酶表达功效的组合物，其中，包含姜黄根提取物和二甲基甲氧基苯并二氢吡喃醇，其中，所述姜黄根提取物和所述二甲基甲氧基苯并二氢吡喃醇的质量份比值为(0.5～3)：1。
  2. 根据权利要求1所述的具有促进抗氧化酶表达功效的组合物，其特征在于，所述姜黄根提取物和所述二甲基甲氧基苯并二氢吡喃醇的质量份比值为(1～2)：1。
  3. 根据权利要求1所述的具有促进抗氧化酶表达功效的组合物，其中，所述姜黄根提取物和所述二甲基甲氧基苯并二氢吡喃醇的质量份比值为1：1。
  4. 权利要求1至3中任一项所述的具有促进抗氧化酶表达功效的组合物在化妆品和/或护肤品中的应用。
  5. 根据权利要求4所述的应用，其中，所述化妆品和/或护肤品施用于皮肤外表面。
  6. 根据权利要求4所述的应用，其中，所述化妆品和/或护肤品选自眼霜、面霜、精华、乳液、面膜精华液中的一种或几种。
  7. 根据权利要求4所述的应用，其中，所述具有促进抗氧化酶表达功效的组合物作为添加剂分散于基质原料中，所述基质原料选自油质原料、粉质原料、胶质原料中的一种或几种。
  8. 根据权利要求4所述的应用，其中，所述化妆品和/或护肤品还包括：表面活性剂、香精与香料、色素、保湿剂、防腐剂、抗氧剂、紫外线吸收剂、螯合剂、收敛剂、助渗剂、营养性添加剂中的一种或几种。
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