WO2023124649A1 - 抗糖化的中药提取物 - Google Patents

抗糖化的中药提取物 Download PDF

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WO2023124649A1
WO2023124649A1 PCT/CN2022/133255 CN2022133255W WO2023124649A1 WO 2023124649 A1 WO2023124649 A1 WO 2023124649A1 CN 2022133255 W CN2022133255 W CN 2022133255W WO 2023124649 A1 WO2023124649 A1 WO 2023124649A1
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chinese medicine
medicine extract
extract
extraction
skin
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PCT/CN2022/133255
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English (en)
French (fr)
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冯春波
陈媛祺
贾海东
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上海家化联合股份有限公司
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/06—Fungi, e.g. yeasts
    • A61K36/07—Basidiomycota, e.g. Cryptococcus
    • A61K36/076—Poria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • 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)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • 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)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
    • A61K36/284—Atractylodes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • 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)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/65—Paeoniaceae (Peony family), e.g. Chinese peony
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • 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)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/71—Ranunculaceae (Buttercup family), e.g. larkspur, hepatica, hydrastis, columbine or goldenseal
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • 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)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/75—Rutaceae (Rue family)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • 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)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/87—Vitaceae or Ampelidaceae (Vine or Grape family), e.g. wine grapes, muscadine or peppervine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • 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)
    • A61K36/898—Orchidaceae (Orchid family)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
    • A61K8/0216—Solid or semisolid forms
    • A61K8/022—Powders; Compacted Powders
    • 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
    • A61K8/9728—Fungi, e.g. yeasts
    • 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]
    • A61K8/9789—Magnoliopsida [dicotyledons]
    • 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
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0014—Skin, i.e. galenical aspects of topical compositions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/18—Antioxidants, e.g. antiradicals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • 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
    • A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
    • A61K2800/59—Mixtures
    • A61K2800/592—Mixtures of compounds complementing their respective functions
    • A61K2800/5922—At least two compounds being classified in the same subclass of A61K8/18
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the invention relates to the fields of natural medicinal chemistry and cosmetics, in particular to a traditional Chinese medicine extract with the effects of anti-glycation, anti-jaundice, whitening and skin-brightening effects, a preparation method and application of the traditional Chinese medicine extract.
  • the hot spot of current research is the theory of non-enzyme glycosylation aging, which is also called the Maillard reaction aging theory.
  • This theory believes that reducing sugars (glucose, galactose, lactose, xylose, fructose) in the skin interact with proteins, nucleic acids,
  • the non-enzymatic reaction of lipids forms glycosylamine with a Schiff base structure, which further undergoes complex rearrangement reactions to become hydroxyl compounds, which combine with amino acids of collagen, and finally form glycosylamine Final products (Advanced Glycation End Products, AGEs).
  • Glycation end products are brown in color, irreversible, cross-linked, not afraid of enzyme damage, not easy to degrade, have special absorption spectrum and fluorescence characteristics, accumulate gradually, and can bind to many cell membrane specific receptors to exert biological effects.
  • AGEs is an important factor leading to collagen cross-linking to aging process. Glycosylation will cause protein cross-linking damage, which can transform the normal protein structure into the structure of aged protein, and also cause the hardening of structural proteins, and the skin is also damaged. It will become fragile and the skin elasticity will be greatly reduced. AGEs can also inhibit the proliferation of fibroblasts and reduce the production of collagen, which is an important factor affecting skin aging.
  • Ultraviolet rays use the characteristics of AGEs to promote oxidative stress, causing oxidative damage to the skin, accelerating photoaging, etc., and wrinkles form and continue Deepening, leading to various aging problems of the skin.
  • Chinese herbal medicine has the functions of regulating yin and yang balance, regulating qi, blood and body fluid, regulating viscera and delaying aging. Therefore, the social and economic benefits of Chinese herbal medicine used in cosmetics development have broad prospects.
  • the present invention unexpectedly finds that the traditional Chinese medicine extracts containing Poria cocos, Tribulus terrestris, Potassium officinalis, Radix Paeoniae Alba, Atractylodes macrocephala, Bletilla striata and Fresh Cortex Bletilla have excellent anti-glycation effect, and can be used as functional additives in cosmetics.
  • the present invention provides an extract of traditional Chinese medicine, prepared from the following raw materials: Poria cocos, Tribulus terrestris, Baishui, Paeoniae officinalis, Atractylodes macrocephala, Bletilla striata and Fresh Cortex Bletilla, wherein Poria cocos: Tribulus terrestris: Potassium striata: Peony: Atractylodes macrocephala: Bletilla striata: The weight ratio of white fresh skin is 1:1-4:4-10:8-10:1-4:1-4:1-4.
  • the Chinese medicine extract has an anti-glycation effect.
  • the Chinese medicine extract is prepared by solvent extraction.
  • the solvent extraction adopts a water extraction and alcohol precipitation method.
  • the solvent extraction uses an alcohol solvent for extraction.
  • the weight ratio of Poria cocos: Tribulus terrestris: Poria chinensis: Paeoniae officinalis: Atractylodes macrocephala: Bletilla striata: Fresh bark is 1:1-2:4-8:8-10:1-4:1-4:1 -2.
  • the present invention provides a composition comprising the Chinese medicine extract of the present invention and polyols.
  • the polyhydric alcohol is selected from: glycerin, diglycerin, butanediol, pentylene glycol, propylene glycol, ethylhexylglycerin, polyethylene glycol, sorbitol or combinations thereof.
  • the polyol is a mixture of butanediol and ethylhexylglycerin in a weight ratio of 1-9:1-9.
  • the present invention relates to a freeze-dried product of a traditional Chinese medicine extract, which is an amorphous powder.
  • the present invention relates to the application of the traditional Chinese medicine extract, the composition, or the freeze-dried product in skin anti-glycation.
  • the present invention relates to an external skin preparation, which comprises the traditional Chinese medicine extract, the composition, or the freeze-dried product.
  • Fig. 1 has shown the HPLC spectrum of the PGG of the traditional Chinese medicine extract of embodiment 4.
  • Figure 2 is the result of immunohistochemical detection.
  • Figure 3 shows the mixed sample of traditional Chinese medicine extracts and polyols after extraction, concentration and purification. From left to right are the pictures of Example 43, Example 121, and Example 126.
  • the purpose of the present invention is based on the compound of ancient Chinese medicine, while synergistic collocation improves the efficacy, the content of active ingredients is increased through the extraction process, the special smell and color of the compound of Chinese medicine are improved, the bioavailability of the compound of Chinese medicine is improved, and it is combined with modern cosmetics technology
  • a traditional Chinese medicine composite product with good anti-skin glycation effect is provided, which can be directly used on the skin, and can also be added to cosmetic formulas as functional additives, greatly improving the efficacy of cosmetic applications, and the cost is low.
  • the present invention combines a plurality of ancient prescriptions of traditional Chinese medicines to obtain a cosmetic prescription with a better combination theory of traditional Chinese medicine.
  • the present invention adopts Poria cocos, Tribulus terrestris, Poria chinensis, Radix peony, Atractylodes macrocephala, Bletilla striata and Fresh Cortex Bletilla to obtain the traditional Chinese medicine extract.
  • the invention utilizes the modern Chinese herbal medicine extraction method and technology to carry out pretreatment on the ancient Chinese medicine prescription.
  • the Chinese herbal medicine extract of the present invention includes pulverizing the raw material of the Chinese herbal medicine before solvent extraction.
  • a high-speed universal pulverizer FW100 type
  • FW100 type can be used to pulverize the mixture of Chinese herbal medicine raw materials.
  • the present invention also utilizes processes such as concentration and purification to improve and enhance the content of active substances in the Chinese medicine compound extract.
  • concentration is achieved by using a Chinese medicinal material extraction and concentration tank (TNH type).
  • the invention utilizes the cosmetic technology to further improve the bioavailability of the skin to the active ingredients of Chinese herbal medicines, and realize the effect of inhibiting AGEs, the terminal product of the glycosylation reaction of the skin.
  • Peony is the dry root of Paeonia lactiflora Pall, a plant of the ranunculaceae family. It contains paeoniflorin, Paeonol, Paeonin, benzoic acid, volatile oil, fatty oil, resin, tannin, sugar, Starch, mucus, protein, ⁇ -sitosterol and triterpenoids, etc.
  • White pomegranate is the dry tuberous root of the grape plant Ampelopsis japonica (Thunb.) Makino, and the root contains mucilage and starch.
  • Tribulus terrestris is the dry ripe fruit of Tribulus terrestris L.
  • the roots and leaves contain saponins, and the aglycones include Diosgenin, Gitogenin, Chlorogenin, and Ruscogenin.
  • Leaves still contain kaempferol (Kaempferol) and a variety of kaempferol glycosides.
  • Atractylodes macrocephala is the dry rhizome of Atractylodes macrocephala Koldz., a plant of the Compositae family, containing 1.4% volatile oil.
  • the main components are Atractylol, Atractylone, etc., and vitamin A.
  • Poria cocos is the dried sclerotia of Polyporaceae fungus Poria cocos (Schw.) Wolf. Tumulosic acid, 3 ⁇ -Hydroxylanostatrienoic acid [3 ⁇ -Hydroxylanosta-7,9(11),24-trien-21-oil acid]. In addition, it still contains gum, chitin, protein, fat, sterol, lecithin, glucose, adenine, histidine, choline, ⁇ -tuckahoe glycan decomposing enzyme, lipase, protease and so on.
  • Baixian is the dried root bark of Rutaceae plant Dictamnus dasycarpus Turcz., containing Dictamnine, Dictamnolactone, Obaculactone, Limonin, Sitosterol, Obacunonic acid , Trigonelline (Trigonelline), Choline (Choline), and Fraxinellone (Fraxinellone). It still contains campesterol, skimmianin, ⁇ -fagarin, and Dasycarpamin. The aerial part contains Psoralen and Xanthotoxin.
  • Bletilla striata is the dried tuber of the orchid plant Bletilla striafa (Thunb.) Reichb.f., and its fresh tuber contains 14.6% moisture, 30.48% starch and 1.5% glucose. It also contains volatile oil and mucus.
  • the root contains Bletilla mannan, which is a glucomannan composed of 4 parts of mannose and 1 part of glucose.
  • the formula is as simple as possible to form a simple formula for balancing and beautifying skin, so as to meet the technical requirements of later extraction and purification technology.
  • Poria cocos, Tribulus terrestris, Bai Po, Paeoniae Atractylodes, Atractylodes macrocephala, Bletilla striata, and Bletilla striata are combined for extraction, and the obtained compound Chinese medicine extracts complement each other to achieve anti-glycation, yellowing, whitening, and anti-glycation. Brightening effect.
  • the weight ratio of the raw materials in the Chinese medicine extract of the present invention is Poria cocos: Tribulus terrestris: Poria cocos: Paeoniae officinalis: Atractylodes macrocephala: Bletilla striata: Fresh bark is 1:1-4:4-10:8-10:1- 4:1-4:1-4.
  • the weight ratio of the raw materials Poria cocos: Tribulus terrestris: Poria cocos: Paeoniae officinalis: Atractylodes macrocephala: Bletilla striata: Fresh bark of Baiji in the Chinese medicine extract of the present invention is 1:4:10:10:1:4:4.
  • the weight ratio of the raw materials Poria cocos: Tribulus terrestris: Poria cocos: Paeoniae officinalis: Atractylodes macrocephala: Bletilla striata: Fresh bark of Baiji in the traditional Chinese medicine extract of the present invention is 1:2:8:10:4:1:2.
  • the weight ratio of the raw materials Poria cocos: Tribulus terrestris: Poria cocos: Paeoniae officinalis: Atractylodes macrocephala: Bletilla striata: Cortex Atractylodis in the Chinese medicine extract of the present invention is 1:1:10:10:4:1:1.
  • the weight ratio of the raw materials Poria cocos: Tribulus terrestris: Poria cocos: Paeoniae officinalis: Atractylodes macrocephala: Bletilla striata: Fresh bark of Baiji in the traditional Chinese medicine extract of the present invention is 1:1:4:10:4:1:1.
  • the weight ratio of the raw materials Poria cocos: Tribulus terrestris: Poria cocos: Paeoniae officinalis: Atractylodes macrocephala: Bletilla striata: Fresh bark of Baiji in the Chinese medicine extract of the present invention is 1:1:4:10:2:1:1.
  • the weight ratio of the raw materials Poria cocos: Tribulus terrestris: Poria cocos: Paeoniae officinalis: Atractylodes macrocephala: Bletilla striata: Fresh bark of Baiji in the Chinese medicine extract of the present invention is 1:2:4:8:4:1:2.
  • the traditional Chinese medicine extract of the invention can be added to cosmetics as functional additives to achieve the effects of anti-glycation, anti-jaundice, whitening and brightening skin tone.
  • the traditional Chinese medicine extract of the present invention is prepared by a solvent extraction method.
  • Common extraction solvents include, but are not limited to: water (such as deionized water), organic solvents (such as ethanol).
  • extraction methods including microwave extraction, supercritical carbon dioxide extraction, ultrasonic extraction, enzyme extraction, etc.
  • the traditional Chinese medicine extract of the present invention is prepared by water extraction and alcohol precipitation.
  • the traditional Chinese medicine extract of the present invention is extracted by boiling with deionized water, and then subjected to alcohol precipitation to obtain the filtrate to obtain the desired Chinese medicine extract.
  • ethanol is used in the alcohol precipitation process.
  • the extraction process of water extraction and alcohol precipitation lasts for 1-24 hours. In some embodiments, the extraction process of water extraction and alcohol precipitation lasts for 12-16 hours.
  • the traditional Chinese medicine extract of the present invention uses alcohol as the extraction solvent.
  • the traditional Chinese medicine extract of the present invention can be extracted with ethanol.
  • the extraction process is carried out under boiling conditions (eg, 80 ⁇ 10° C.).
  • 70% by volume ethanol can be used for extraction.
  • the alcohol extraction process lasts 2-4 hours.
  • the present invention also relates to the composition comprising the traditional Chinese medicine extract and polyalcohol.
  • polyol used in the present invention refers to water-soluble polyols with two or more hydroxyl groups in the molecule, including but not limited to: glycerin, diglycerin, butanediol, pentanediol, propylene glycol, ethylhexylglycerin, poly Ethylene glycol, sorbitol, etc.
  • Polyols are often used in cosmetics as humectants, absorbing moisture from the environment, replenishing it into the skin, and increasing the moisture content of the skin surface. Polyols may be used alone, or they may be used in combination. For example, 2 or more polyols may be used in combination.
  • the traditional Chinese medicine extract of the present invention is combined with a mixture of pentylene glycol and propylene glycol in a weight ratio of 9:1. In some embodiments, the traditional Chinese medicine extract of the present invention is combined with a mixture of polyethylene glycol and caprylyl glycol in a weight ratio of 5:5.
  • the invention also relates to the freeze-dried product of the Chinese medicine extract.
  • the lyophilized product can be an amorphous powder.
  • freeze-drying is performed using a vacuum freeze dryer FD-2 from Beijing Boyikang Experimental Instrument Co., Ltd.
  • the freeze-drying setting is -80 ⁇ -40°C
  • the vacuum degree is 1-30Pa
  • the time is 8 hours
  • the sublimation drying setting is -20 ⁇ +20°C
  • the vacuum degree is 1-30Pa
  • the time is 6 hours
  • the water content is controlled at Between 0 and 6%
  • the analytical drying setting is -10 to +20°C
  • the vacuum degree is 1 to 30Pa
  • the time is 3 hours
  • the water content is controlled between 0 and 6%.
  • the present invention also relates to an external skin preparation, which comprises the Chinese medicine extract of the present invention, the composition of the Chinese medicine extract and polyol, or the freeze-dried product of the Chinese medicine extract, and cosmetic, dermatological or pharmaceutical acceptable excipients.
  • the dosage of the Chinese medicine extract is 0.001-50% (w/w), preferably 1-20% (w/w), more preferably 2-10% (w/w), most preferably 3-8% (w/w).
  • freeze-dried product of the extract of the traditional Chinese medicine can also be used alone.
  • the skin external preparation composition is a general term for all ingredients that are usually used on the outside of the skin, and may be, for example, a cosmetic composition or a pharmaceutical composition.
  • the cosmetic composition can be basic cosmetics, facial makeup cosmetics, body cosmetics, hair care cosmetics, etc.
  • Such external preparations for skin can be formulated into any suitable product form.
  • product forms include, but are not limited to, aerosol sprays, creams, lotions, solids, liquids, dispersions, foams, gels, lotions, mousses, ointments, powders, patches, pomades, solutions , pump sprays, sticks, masks, and wipes.
  • Such external preparations for skin can be conveniently used in the preparation or as cosmetic, dermatological or pharmaceutical topical products by various methods well known in the art.
  • the skin external preparation composition of the present invention may include one or more of the following ingredients: antiallergic agents, antimicrobial agents, antioxidants, chelating agents, coloring agents, depigmenting agents, emollients, emulsifiers, exfoliating agents, active ingredients Film agent, fragrance, humectant, insect repellant, lubricant, pharmaceutically active agent, moisturizer, photostabilizer, preservative, skin care agent, skin penetration enhancer, sunscreen agent, stabilizer, surfactant, thickener agents, viscosity modifiers, vitamins or any combination thereof.
  • the cosmetically, dermatologically or pharmaceutically acceptable excipients that can be used in the skin external preparation composition of the present invention are water phase, oil phase, gel, wax-in-water emulsion, oil-in-water emulsion or water-in-oil in the form of emulsion.
  • the aqueous phase is a mixture of one or more water-soluble or dispersible components, which may be liquid, semi-solid or solid at room temperature (25°C).
  • the vehicle includes or can be in the form of a suspension, dispersion or solution in aqueous or aqueous-alcoholic vehicles, which may contain thickening or gelling agents. Those skilled in the art can select a suitable product form and components contained therein based on the knowledge acquired by those skilled in the art.
  • Said composition may comprise an aqueous phase which may contain water or a mixture of water and at least one hydrophilic organic solvent, such as an alcohol, especially containing 2 to 5 carbon atoms straight or branched lower monohydric alcohols such as ethanol or propanol; polyols such as propylene glycol, sorbitol, glycerin, panthenol or polyethylene glycol and mixtures thereof.
  • a hydrophilic organic solvent such as an alcohol, especially containing 2 to 5 carbon atoms straight or branched lower monohydric alcohols such as ethanol or propanol
  • polyols such as propylene glycol, sorbitol, glycerin, panthenol or polyethylene glycol and mixtures thereof.
  • composition of the invention when the composition of the invention is in the form of an emulsion, the composition may also optionally comprise a surfactant.
  • the composition may also comprise film-forming polymers such as polyurethanes, polyacrylic acid homo- or copolymers, polyesters, hydrocarbon-based resins and/or silicone resins.
  • film-forming polymers such as polyurethanes, polyacrylic acid homo- or copolymers, polyesters, hydrocarbon-based resins and/or silicone resins.
  • the polymer can be dissolved or dispersed in a cosmetically acceptable vehicle and optionally combined with a plasticizer.
  • composition of the invention may also comprise an oily phase comprising oil-soluble or oil-dispersible components which are liquid at room temperature (25° C.) and/or substances which are oily or waxy at room temperature, such as Waxes, semi-solids, gums and mixtures thereof.
  • oily phase may also contain organic solvents.
  • suitable oily substances include: hydrocarbon-based oils of animal origin, such as perhydrosqualene; hydrocarbon-based vegetable oils, such as liquid triglycerides of C4-10 fatty acids, such as heptanoic acid or caprylic triglycerides, or oils such as sunflower oil, corn oil, soybean oil, grapeseed oil, castor oil, avocado oil, caprylic/capric triglycerides, jojoba oil; straight from mineral or synthetic origin Chain or branched hydrocarbons, such as liquid paraffin and its derivatives, petrolatum; synthetic esters and ethers, especially esters of fatty alcohols, such as isopropyl myristate, 2-ethylhexyl palmitate, hard 2-Octyldodecyl fatty acid, isostearyl isostearate; hydroxylated esters such as isostearyl lactate, octyl hydroxystearate, octyl hydroxystea
  • composition of the present invention may further comprise any components commonly used in the field of cosmetics. These components include preservatives, aqueous phase thickeners (extract biopolymers, synthetic polymers) and fatty phase thickeners, fragrances, hydrophilic and lipophilic active agents and mixtures thereof.
  • compositions according to the invention may also comprise an additional particulate phase, which may be pigments and/or pearlescent agents and/or fillers used in cosmetic compositions.
  • Pigments may be present in the composition, suitable inorganic pigments include titanium oxide, zirconium oxide and cerium oxide as well as zinc oxide, iron oxide and iron blue; suitable organic pigments include barium, strontium, calcium and aluminum lakes and carbon black.
  • Pearlescent agents may be present in the composition, suitable pearlescent agents include mica coated with titanium oxide, iron oxide or natural pigments.
  • Fillers may be present in the composition, suitable fillers include talc, silicon dioxide, zinc stearate, mica, kaolin, nylon powder, polyethylene powder, Teflon, starch, boron nitride, copolymers Microspheres, such as silicone resin microspheres.
  • the oily phase of the compositions of the present invention may comprise one or more waxes, gums or mixtures thereof.
  • Waxes include hydrocarbon-based, fluoro, and/or silicone waxes, and may be derived from vegetable, mineral, animal, and/or synthetic sources. Suitable waxes include beeswax, carnauba wax, candelilla wax, paraffin wax, microcrystalline wax, ozokerite wax; synthetic waxes include polyethylene wax, silicone wax containing C16-45. Gums are typically polydimethylsiloxane or sodium carboxymethylcellulose or extracts, and semi-solid materials are typically hydrocarbon-based compounds such as lanolin and its derivatives.
  • compositions of the invention may be formulated in any suitable product form.
  • product forms include, but are not limited to, aerosol sprays, creams, lotions, solids, liquids, dispersions, foams, gels, lotions, mousses, ointments, powders, patches, pomades, solutions , pump sprays, sticks, masks, and wipes.
  • the compositions of the present invention may be conveniently used in the preparation or as cosmetic, dermatological or pharmaceutical topical products by various methods well known in the art.
  • the skin external preparation composition of the present invention may include one or more of the following ingredients: antiallergic agents, antimicrobial agents, antioxidants, chelating agents, coloring agents, depigmenting agents, emollients, emulsifiers, exfoliating agents, active ingredients Film agent, fragrance, humectant, insect repellant, lubricant, pharmaceutically active agent, moisturizer, photostabilizer, preservative, skin care agent, skin penetration enhancer, sunscreen agent, stabilizer, surfactant, thickener agents, viscosity modifiers, vitamins or any combination thereof.
  • the raw materials of medicinal materials used in the following examples were all purchased from “Anhui Bozhou Jingwan Traditional Chinese Medicine Decoction Pieces Factory", and the incoming material was in the form of "Chinese Herbal Medicine Decoction Pieces”; the biological reagents used were all purchased from Shanghai Jiahua Biotechnology Co., Ltd.
  • Embodiment 1 the preparation of Chinese medicine extract
  • Embodiment 2 the preparation of Chinese medicine extract
  • Embodiment 3 the preparation of Chinese medicine extract
  • Embodiment 4 the preparation of Chinese medicine extract
  • Embodiment 5 the preparation of Chinese medicine extract
  • Embodiment 6 the preparation of Chinese medicine extract
  • Embodiment 7 the preparation of Chinese medicine extract
  • Embodiment 8 the preparation of Chinese medicine extract
  • Embodiment 9 the preparation of Chinese medicine extract
  • Embodiment 10 the preparation of Chinese medicine extract
  • Embodiment 11 the preparation of Chinese medicine extract
  • Embodiment 12 the preparation of Chinese medicine extract
  • Embodiment 13 the preparation of Chinese medicine extract (comparative example)
  • Embodiment 14 the preparation of Chinese medicine extract (comparative example)
  • Embodiment 15 the preparation of Chinese medicine extract (comparative example)
  • Embodiment 16 the preparation of Chinese medicine extract (comparative example)
  • Embodiment 17 the preparation of Chinese medicine extract (comparative example)
  • Embodiment 18 the preparation of Chinese medicine extract (comparative example)
  • Embodiment 19 the preparation of Chinese medicine extract (comparative example)
  • Embodiment 20 the preparation of Chinese medicine extract (comparative example)
  • Embodiment 21 the preparation of Chinese medicine extract (comparative example)
  • Embodiment 22 the preparation of Chinese medicine extract (comparative example)
  • Embodiment 23 the preparation of Chinese medicine extract (comparative example)
  • Embodiment 24 the preparation of Chinese medicine extract (comparative example)
  • Embodiment 25 the preparation of Chinese medicine extract (comparative example)
  • Embodiment 26 the preparation of Chinese medicine extract (comparative example)
  • Embodiment 27 the preparation of Chinese medicine extract (comparative example)
  • Embodiment 28 Preparation of Chinese medicine extract
  • Embodiment 29 Preparation of Chinese medicine extract
  • Embodiment 30 the preparation of Chinese medicine extract
  • Embodiment 39 Preparation of Chinese medicine extract
  • Test example 1 detection of active substance content
  • the chemical structure of the active ingredient of the drug is the key to the percutaneous penetration of the drug.
  • Fat-soluble drugs easily penetrate the stratum corneum, skin barrier, and cell membrane, but diffuse slowly in the active epidermis due to the relatively high water content.
  • the water-soluble component drugs pass through the stratum corneum cell membrane and the skin barrier at a slower rate, but diffuse faster in the active epidermis.
  • attention should be paid to the molecular structure of the ingredients as much as possible to choose pharmaceutical ingredients with moderate oil/water distribution, or to use water-soluble and fat-soluble ingredients together, often to get better results.
  • 1,2,3,4,6-O-Galloyl Glucose is a polyphenolic compound composed of 5 gallic acids and 1 glucose through an ester bond, with a relative molecular weight of 940 and a good Hydrophilic and lipophilic, with strong physiological activities, such as anti-tumor, anti-oxidation, anti-leukemia, anti-allergic, hypoglycemic and other functions, it exists in a variety of Chinese medicinal materials.
  • the chromatographic conditions and system suitability test used C18 general-purpose Eclipse XDB-C18 (4.6x250mm, 5 ⁇ m) chromatographic column; acetonitrile-0.1% dilute phosphoric acid (25:75) was used as the mobile phase; after gradient elution, the detection wavelength was 230nm; The column temperature is 35 °C; the flow rate is 0.8 mL/min; the injection volume is 10 ⁇ L.
  • the number of theoretical plates should not be less than 200,000 based on the peak value of 1,2,3,4,6-O-galloylglucose (PGG).
  • Preparation of the reference substance solution Accurately weigh the PGG reference substance dried to constant weight, add methanol to make a mixed solution containing 0.2mg per 1mL, and shake well.
  • the Chinese medicine extracts of Examples 1 to 6 obtained a higher amount of PGG, especially in Example 2, which reached 0.7913mg/ml, which was 96% higher than that of Example 6. %.
  • the content of the active ingredient PGG is generally low, especially in Example 8 with the same proportion, compared with Example 2 For example, the content is about 13.7% lower; similarly, in the water extraction and alcohol precipitation process of Examples 1-6, even with the same ratio and compatibility, the overall PGG content is higher than that of Examples 7-12 alcohol extraction process.
  • Test Example 2 Comparison of anti-glycation efficacy (detection index AGEs)
  • the ExVivo tissue (provided by Guangdong Boxi Biotechnology Co., Ltd.) was obtained, and the subcutaneous fat tissue was removed by using a common tool, a roller dermatome, to thin the fat end of the isolated skin tissue block to achieve the purpose.
  • the skin tissue blocks after subcutaneous fat removal were stored at low temperature (-20°C) and frozen for future use.
  • Penicillin-streptomycin (10,000U/ml) mixed solution is also called double antibody, that is, a solution containing 10,000 units/ml penicillin and 10,000 ⁇ g/ml streptomycin, and the manufacturer is Thermo Fisher Scientific (China) Co., Ltd. , That is, prepare 100ml of PBS solution and add 1ml of double antibody solution, the purpose is to play a combined role in resisting Gram-positive bacteria and Gram-negative bacteria, and prevent skin tissue blocks from being contaminated by bacteria.
  • Phosphate buffered saline solution (PBS) is a kind of balanced salt solution, the manufacturer is Beijing Solaibao Technology Co., Ltd., P1020-500ml 1xPBS buffer solution (pH7.2-7.4).
  • Tissue blocks with a diameter of 4-6 mm were taken and frozen and thawed five times in liquid nitrogen and a 37°C water bath. Place the isolated skin tissue block in a 24-well plate, add 1ml of PBS containing double antibody, and incubate with CO 2 for 18 ⁇ 2h.
  • the saccharification reaction is carried out according to the following steps:
  • UVA irradiation uses a UVA ultraviolet phototherapy instrument, the manufacturer Shanghai SIGMA High-Tech Co., Ltd., according to the irradiation dose of 10J/cm 2 UVA, 72 hours each time;
  • the saccharification reaction working solution is methylglyoxal working solution, that is, according to the ratio of 1:13000, dilute to 0.5mM with PBS solution containing double antibody, methyl ethyl Dialdehyde manufacturer is Sigma, M0252 model
  • add the newly prepared saccharification reaction working solution namely the aminoguanidine sulfate of 3mM concentration as positive control, 3 weight % the extract that embodiment 1 ⁇ 120 obtains, post-treatment or post-treatment
  • UVA irradiation was carried out for 3 times. After the 4th time, the isolated skin was cleaned and then transferred to a new saccharification reaction working solution. Steps 3) and 4) were repeated for a total of 21 days.
  • the isolated skin tissue was transferred to a 1.5mL EP tube, fixed with 4% paraformaldehyde, embedded, sectioned, and immunohistochemically stained with AGEs.
  • IHC immunohistochemistry
  • PBS buffer was purchased from Thermo Fisher Scientific (China) Co., Ltd., model eBioscience IHC solution.
  • SP kit manufactured by Fuzhou Maixin Company, model E-EL-0067C
  • DAB staining select 3 fields of view for each slice under an inverted Olympus microscope calculate data.
  • Relative rate of improvement (%) (negative control-relative average value of embodiment)/negative control x 100%
  • Example 1 1.52 0.02 10.59
  • Example 2 1.59 0.04 6.47
  • Example 3 1.56 0.05 8.24
  • Example 4 1.56 0.05 8.24
  • Example 5 1.54 0.04 9.41
  • Example 6 1.57 0.05 7.65
  • Example 7 1.64 0.04 3.53
  • Example 8 1.62 0.06 4.71
  • Example 9 1.65 0.06 2.94
  • Example 10 1.61 0.05 5.29
  • Example 11 1.59 0.05 6.47
  • Example 12 1.60 0.04 5.88
  • Example 13 1.70 0.02 0.00
  • Example 14 1.72 0.01 -1.18
  • Example 15 1.74 0.01 -2.35
  • Example 16 1.69 0.01 0.59
  • Example 17 1.68 0.01 1.18
  • Example 18 1.66 0.01 2.35
  • Example 19 1.63 0.01 4.12
  • Example 20 1.65 0.03 2.94
  • Example 21 1.79 0.04 -5.29
  • Example 22 1.82 0.03 -7.06
  • Example 23 1.70 0.05 0.00
  • Example 24 1.68 0.07 1.18
  • Example 25 1.69 0.05 0.59
  • Example 26 1.69 0.06 0.59
  • Example 27 1.60 0.05 5.88
  • Example 28 1.57 0.07 7.65
  • Example 29 1.49 0.07 12.35
  • Example 30 1.57 0.04 7.65
  • Example 31 1.66 0.03 2.35
  • Example 32 1.72 0.03 -1.18
  • Example 33 1.69 0.04 0.59
  • Example 34 1.65 0.05 2.94
  • Example 35 1.63 0.05 4.12
  • Example 36 1.66 0.04 2.35
  • Example 37 1.68 0.04 1.18
  • Example 38 1.69 0.03 0.59
  • Example 39 1.72 0.02 -1.18
  • Example 40 1.68 0.02 1.18
  • Example 41 1.29 0.01
  • Example 46 1.39 0.03 18.24 Example 47 1.37 0.04 19.41 Example 48 1.41 0.03 17.06 Example 49 1.45 0.05 14.71
  • Example 50 1.58 0.07 7.06
  • Example 51 1.40 0.05 17.65
  • Example 52 1.57 0.06 7.65
  • Example 53 1.53 0.05 10.00
  • Example 54 1.52 0.07 10.59
  • Example 55 1.55 0.07 8.82
  • Example 56 1.51 0.04 11.18
  • Example 58 1.60 0.03 5.88
  • Example 59 1.61 0.04 5.29
  • Example 60 1.62 0.05 4.71
  • Example 61 1.66 0.05 2.35
  • Example 62 1.53 0.04 10.00
  • Example 63 1.65 0.04 2.94
  • Example 64 1.59 0.01 6.47
  • Example 65 1.54 0.01 9.41
  • Example 68 1.46 0.01 14.12
  • Example 69 1.47 0.01 13.53
  • Example 70 1.48 0.03 12.94
  • Example 75 1.55 0.05 8.82
  • Example 76 1.59 0.06 6.47
  • Example 77 1.63 0.05 4.12
  • Example 78 1.72 0.07 -1.18
  • Example 79 1.69 0.07 0.59
  • Example 80 1.51 0.04 11.18
  • Example 81 1.49 0.03 12.35
  • Example 82 1.56 0.03 8.24
  • Example 83 1.55 0.04 8.82
  • Example 84 1.43 0.05 15.88
  • Example 85 1.39 0.05 18.24
  • Example 86 1.48 0.04 12.94
  • Example 87 1.55 0.04 8.82
  • Example 88 1.48 0.01 12.94
  • Example 89 1.57 0.01 7.65
  • Example 90 1.64 0.01 3.53
  • Example 91 1.62 0.01 4.71
  • Example 92 1.65 0.01 2.94
  • Example 93 1.66 0.01 2.35
  • Example 94 1.69 0.03 0.59
  • Example 95 1.70 0.04 0.00
  • Example 96 1.72 0.03 -1.18
  • Example 97 1.70 0.05 0.00
  • Example 104 1.47 0.05 13.53
  • Example 105 1.57 0.07 7.65
  • Example 106 1.63 0.05 4.12
  • Example 107 1.62 0.06 4.71
  • Example 108 1.65 0.05 2.94
  • Example 109 1.69 0.07 0.59
  • Example 110 1.62 0.07 4.71
  • Example 111 1.47 0.04 13.53
  • Example 112 1.49 0.03 12.35
  • Example 113 1.53 0.03 10.00
  • Example 114 1.57 0.04 7.65
  • Example 115 1.54 0.05 9.41
  • Example 116 1.48 0.03 12.94
  • Example 117 1.43 0.04 15.88
  • Example 118 1.47 0.02 13.53
  • Example 119 1.40 0.03 17.65
  • Example 120 1.51 0.02 11.18
  • the first thing that needs to be studied for the core efficacy of traditional Chinese medicine cosmetics is the transdermal absorption and utilization of active ingredients.
  • the keratinocytes are arranged more closely, which reduces the effective penetration area of the skin and the space for substances to penetrate the skin, thereby reducing the degree and efficiency of drug transdermal absorption.
  • the first thing to do for functional cosmetics is to restore the water content of the skin to a normal level or in a moist state.
  • the water content of the stratum corneum is 10%.
  • the other layers of the epidermis are 10% to 47%.
  • the compact keratinocytes form a porous and plump state, increasing the effective area of the skin and the contact cross-section ,
  • the transdermal absorption rate of the drug is greatly improved, and the drug will play a better role in the skin moisturizing environment.
  • ceramide and cholesterol have a great shielding effect on cells, which affects the absorption and utilization of drugs; due to the large molecular weight of hyaluronic acid (HA), its molecular sieve effect is not conducive to the penetration and absorption of active ingredients of drugs, especially the formulation system When pH ⁇ 5, the network cross-linking of hyaluronic acid molecules will be strengthened, increasing the resistance of drug diffusion.
  • the glycation reaction in the skin occurs deep in the epidermis and dermis. If any functional active substance cannot pass through the skin barrier, it will not be able to reach the glycation site and exert its actual effect.
  • the extract of the present application can realize transdermal absorption, and help whitening substances penetrate and be utilized by skin cells to achieve the effect of anti-glycation.
  • Example 21 Example 22, and Example 39; but there are exceptions, that is, Example 29, the anti-glycation improvement rate reached 12.35%, which is a relatively high improvement performance.
  • the anti-glycation improvement rate of the extract obtained from single-flavored Chinese herbal medicine is not high no matter whether it is prepared by water extraction and alcohol precipitation or alcohol precipitation.
  • From Examples 41 to 46 are the post-treatment products of the extracts of Examples 1 to 6.
  • the production of the extracts of Examples 1 to 6 through the water extraction and alcohol precipitation process is greatly improved.
  • the improvement rate of the obtained extract in inhibiting AGEs, such as embodiment 43 is improved to 28.82%, embodiment 42 is improved to 25.29%, embodiment 41 is improved to 24.12%, embodiment 44 is improved to 19.41%, and the average improvement ratio exceeds 10%, indicating that the combination of the selected two alcohols can greatly improve the anti-glycation effect of the traditional Chinese medicine extract.
  • Examples 47-52 are the post-treatment products of the extracts of Examples 7-12, and also by compounding with specific polyols, it also greatly improves the production of the extracts of Examples 7-12 through the alcohol extraction process.
  • the improvement rate of the obtained extracts in inhibiting AGEs, such as Example 47 has also increased to 19.41%, Example 48 has increased to 17.06%, and Example 51 has increased to 17.65%, which is more than 10% higher than that of the extracts of Examples 7-12. %. It shows that under the alcohol extraction process, the anti-glycation effect of the active substance can be greatly improved by adding butanediol and ethylhexylglycerin.
  • the main functional substances in Chinese herbal medicine compound additives are flavonoids, polyphenols, polysaccharides, etc.
  • the distribution and content of the specific components mostly depend on the extraction method and means, and because most substances are not easy to preserve, improper storage is prone to oxidation, decomposition, volatilization, Deterioration and other problems lead to the loss of functional substances.
  • the advanced freeze-drying technology is used in the later stage of the preparation process, so that the active ingredients in the extract can be better retained, and the original flavor of the active ingredients can be retained to a greater extent, especially the heat-sensitive active ingredients. It operates in the absence of oxygen, therefore, some easily oxidized active ingredients (such as oils) can also be protected.
  • Examples 1-40 were processed by freeze-drying technology to obtain the freeze-dried products of Examples 81-120, which can avoid the loss of active ingredients caused by improper storage of active ingredients due to various possible reasons, thus affecting the anti-glycation The effect of the reaction.
  • the freeze-dried products that is, the inhibition of AGEs in Examples 81-120, have been tested to inhibit glycosylated AGEs. From the improvement rate of the results in Table 2, they are generally equivalent to the test results of the extract itself. Individual implementations The test results of the examples have been improved, such as Example 85 improves the improvement rate of AGEs to 18.24%, and Example 102 improves the improvement rate of AGEs to 15.88%, showing a certain degree of growth. It shows that the activity of the extract itself and the effect of inhibiting glycosylated AGEs can be better preserved during the entire storage process of the extract from the extract to the freeze-drying process.
  • Figure 2 shows the results of three parallel experiments on the experimental samples of Example 42 and Example 43.
  • the negative control showed a negative improvement of -70%. It can be seen from the figure that the degree and color of the glycosylation reaction are darker.
  • the positive control improved by 22.94%, but for the experimental samples Example 42 and Example 43, the color of the picture is obviously light, indicating that the effect of inhibiting AGEs in these two examples is better, reaching 25.29% respectively and 28.82%.
  • Embodiment 121 ⁇ 125 Post-processing of Chinese medicine extract (comparative example)
  • Embodiment 126 ⁇ 130 Post-processing of Chinese medicine extract (comparative example)
  • Embodiment 131 polyol combination ratio (comparative example)
  • Embodiment 132 polyol combination ratio (comparative example)
  • Embodiment 133 Polyol combination ratio (comparative example)
  • Test Example 3 Comparison of anti-glycation efficacy (detection index AGEs)
  • the ExVivo tissue (provided by Guangdong Boxi Biotechnology Co., Ltd.) was obtained, and the subcutaneous fat tissue was removed by using a common tool, a roller dermatome, to thin the fat end of the isolated skin tissue block to achieve the purpose.
  • the skin tissue blocks after subcutaneous fat removal were stored at low temperature (-20°C) and frozen for future use.
  • Penicillin-streptomycin (10,000U/ml) mixed solution is also called double antibody, that is, a solution containing 10,000 units/ml penicillin and 10,000 ⁇ g/ml streptomycin, and the manufacturer is Thermo Fisher Scientific (China) Co., Ltd. , That is, prepare 100ml of PBS solution and add 1ml of double antibody solution, the purpose is to play a combined role in resisting Gram-positive bacteria and Gram-negative bacteria, and prevent skin tissue blocks from being contaminated by bacteria.
  • Phosphate buffered saline solution (PBS) is a kind of balanced salt solution, the manufacturer is Beijing Solaibao Technology Co., Ltd., P1020-500ml 1xPBS buffer solution (pH7.2-7.4).
  • Tissue blocks with a diameter of 4-6 mm were taken and frozen and thawed five times in liquid nitrogen and a 37°C water bath. Place the isolated skin tissue block in a 24-well plate, add 1ml of PBS containing double antibody, and incubate with CO 2 for 18 ⁇ 2h.
  • the saccharification reaction is carried out according to the following steps:
  • UVA irradiation uses a UVA ultraviolet phototherapy instrument, the manufacturer Shanghai SIGMA High-Tech Co., Ltd., according to the irradiation dose of 10J/cm 2 UVA, 72 hours each time;
  • the saccharification reaction working solution is methylglyoxal working solution, that is, according to the ratio of 1:13000, dilute to 0.5mM with PBS solution containing double antibody, methyl ethyl
  • the dialdehyde manufacturer is Sigma, M0252 model
  • add the newly configured saccharification reaction working solution i.e. aminoguanidine sulfate of 3mM concentration as a positive control, 3% by weight of the extract or post-treatment obtained in Examples 121-133, repeat Steps 2) to 4);
  • UVA irradiation was carried out for 3 times. After the 4th time, the isolated skin was cleaned and then transferred to a new saccharification reaction working solution. Steps 3) and 4) were repeated for a total of 21 days.
  • the isolated skin tissue was transferred to a 1.5mL EP tube, fixed with 4% paraformaldehyde, embedded, sectioned, and immunohistochemically stained with AGEs.
  • IHC immunohistochemistry
  • PBS buffer was purchased from Thermo Fisher Scientific (China) Co., Ltd., model eBioscience IHC solution.
  • SP kit manufactured by Fuzhou Maixin Company, model E-EL-0067C
  • DAB staining select 3 fields of view for each slice under an inverted Olympus microscope calculate data.
  • Relative rate of improvement (%) (negative control-embodiment relative mean value)/negative control x 100%.
  • Example 121 1.36 0.05 20.00
  • Example 122 1.41 0.05 17.06
  • Example 123 1.38 0.04 18.82
  • Example 124 1.31 0.05 22.94
  • Example 125 1.39 0.04 18.24
  • Example 126 1.41 0.06 17.06
  • Example 128 1.41 0.01 17.06
  • Example 129 1.47 0.01 13.53
  • Example 130 1.40 0.01 17.65
  • Example 131 1.69 0.03 0.59
  • Example 132 1.72 0.02 -1.18
  • Example 133 1.67 0.04 1.76
  • Examples 121 to 125 are post-treatment products of pentylene glycol + propylene glycol
  • Examples 126 to 130 are polyethylene glycol
  • Table 4 The horizontal comparison between the post-treatment product of alcohol + caprylyl glycol and the extract obtained by the water extraction and alcohol precipitation preparation process on the inhibition of AGEs, the terminal product of glycosylation reaction, is shown in Table 4. It can be seen from the table that the products obtained from the post-treatment of Examples 1-5 by using the combination of butanediol+ethylhexylglycerin have the highest improvement rate on AGEs, the terminal product of the glycosylation reaction.
  • pentylene glycol + propylene glycol, and polyethylene glycol + caprylyl glycol improved the anti-glycation effect of Chinese herbal extracts to a certain extent, indicating that different combinations of polyols also play an important role in the anti-glycation effect of the active substances of Chinese herbal medicine compounds. enhancement. While there is a certain correlation with the above-mentioned higher content of PGG, the addition of different types of polyols also causes differences in the utilization rate of active ingredients of Chinese herbal medicines by cells.
  • the skin external preparation is preferably a cosmetic composition such as lotion, essence, cream and the like.
  • the weight percentage of the compound extract in the skin external preparation is 0.001%-50% (w/w).
  • the preferred weight percentage is 1%-20% (w/w).
  • a more preferred weight percentage is 2%-10% (w/w).
  • the most preferred weight percentage is 3%-8% (w/w).
  • Examples 82-121 in the present invention can also be used alone as low-temperature freeze-dried products.
  • the above-mentioned cosmetics prepared by using the composition of the present invention have the same or better use effect as that of the commercially available products.
  • the present invention illustrates the detailed methods of the present invention through the above examples, but the present invention is not limited to the above detailed methods, that is, it does not mean that the present invention must rely on the above detailed methods to be implemented.
  • Those skilled in the art should understand that any improvement of the present invention, the equivalent replacement of each raw material of the product of the present invention, the addition of auxiliary components, the selection of specific methods, etc., all fall within the scope of protection and disclosure of the present invention.

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Abstract

一种用于皮肤抗糖化的中药提取物，通过以下原料制备得到：茯苓、蒺藜、白蔹、芍药、白术、白芨和白鲜皮，其中茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮的重量比为1:1-4:4-10:8-10:1-4:1-4:1-4，采用水提醇沉或醇溶剂提取的方法得到。所述中药提取物与多元醇的组合物以及所述中药提取物的冻干产品。所述提取物、组合物或冻干产品在制备皮肤抗糖化产品中的用途，和包含所述提取物、组合物或冻干产品的皮肤外用剂。

Description

抗糖化的中药提取物 技术领域
本发明涉及天然药物化学领域和化妆品领域，具体涉及一种具有抗糖化、祛黄、美白、提亮肤色功效的中药提取物，该中药提取物的制备方法以及应用。
背景技术
中医药是中华民族的瑰宝，数千年来为中华民族的繁荣昌盛做出了不可磨灭的贡献，并越来越受到世界人民的关注和重视。但由于传统中医药理论与现代科学理论脱节以及中药研究手段相对落后，使中药资源的开发和利用受到极大的影响和限制。中药的开发目前主要局限于从现有中药中寻找有效成分，并发展各种提取的手段富集和优化有效成分。而以现有中药资源为基础提升中药的开发潜力、应用效率和新的应用场景，目前尚未引起人们的广泛重视，特别是将传统中药的优势与现代化妆品组方技术相结合，实现中药的现代化是促进中药发展的重要手段。
现代中草药化妆品以其秉承中医智慧，处处体现整体观念，采用的符合中草药组分不仅加强了中草药化妆品的功能性，还使得其具有一专多能的特性，并且具有天然属性安全性高，毒副作用小、易于利用吸收等特点具有广阔的开发应用前景。随着社会的进步和科技的发展，人们对肌肤的问题的认识不断加深，如衰老的机制、美白的机制、敏感的机制等，人们对抗衰老，永葆青春容颜的愿望也日趋强烈。
当今研究的热点非酶糖基化衰老学说，该学说又称为美拉德反应衰老学说，该学说认为肌肤内还原性糖(葡萄糖、半乳糖、乳糖、木糖、果糖)与蛋白质、核酸、脂质发生的非酶反应形成席夫碱(Schiff)结构的糖基化氨(glycosylamine)，进一步发生复杂的重排反应变成羟基类化合物，与胶原蛋白的氨基酸相结合，最后形成糖基化终产物(Advanced Glycation End Products， AGEs)。糖基化终产物(AGEs)呈棕褐色、不可逆、交联、不怕酶破坏、不易降解、有特别的吸收光谱及荧光特性、逐渐累积，可与许多细胞膜特异性受体结合发挥生物学效应。AGEs是导致胶原蛋白交联至衰老过程中的重要影响因素，糖基化会造成蛋白质的交联损伤，能使正常的蛋白结构转变成老年蛋白的结构，还会造成结构蛋白的硬化，皮肤也会因此变得脆弱，皮肤弹性大幅下降。AGEs也会抑制成纤维细胞的增殖，减少胶原蛋白的生成，是皮肤衰老的重要影响因素，紫外线又利用AGEs的特性促进氧化应激反应，造成肌肤氧化损伤，加速光老化等，皱纹形成并不断加深，导致皮肤各种衰老问题。
中医理论体系与分子生物学虽然属于两种不同的思想体系，但研究生命活动的物质基础是一致的，是人类关于生命与健康在不同程度上的认识和探索的产物。比如中医理论认为衰老机理是“气血不和，百病乃变化而生”的观点，即气虚血瘀是衰老的根本原因。现在医学认为气虚血瘀表现为微循环不畅、血液流动性不佳、肌肤内部的代谢不佳造成的。因此，从分子水平研究中医理论及中医药，并将中医理论及现代中医药的其他新成果应用到现代生物学技术中去，对推动中西医结合、促进医药在美容护肤方面的发展具有重要的意义。从现代医学研究来看，中草药中含有丰富的蛋白质、脂质、氨基酸、多糖类、果胶、维生素、微量元素、有机酸、生物碱、皂苷等营养物质，具有促进细胞增殖代谢，改善皮肤微循环，增强机体抗氧化能力，抗击紫外线损伤，抑制羰基化反应，保湿和皮肤屏障修护等护肤功效。中医的观点则对中草药调理阴阳平衡，调理气血津液，调理脏腑和延缓衰老等功能，故中草药应用于化妆品开发的社会和经济效益前景广阔。
本发明意外地发现，包含茯苓、蒺藜、白蔹、芍药、白术、白芨、白鲜皮的中药提取物具有优异的抗糖化作用，能够作为功效添加剂应用于化妆品中。
发明内容
一方面，本发明提供了一种中药提取物，通过以下原料制备得到：茯苓、蒺藜、白蔹、芍药、白术、白芨和白鲜皮，其中茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮的重量比为1:1-4:4-10:8-10:1-4:1-4:1-4。
在优选的实施方式中，中药提取物具有抗糖化作用。
在优选的实施方式中，中药提取物通过溶剂提取制备得到。在优选的实施方式中，所述溶剂提取采用水提醇沉方法。在优选的实施方式中，所述溶剂提取采用醇溶剂进行提取。在优选的实施方式中，茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮的重量比为1:1-2:4-8:8-10:1-4:1-4:1-2。
另一方面，本发明提供了一种组合物，所述组合物包含本发明的中药提取物以及多元醇。在优选的实施方式中，多元醇选自：甘油、双甘油、丁二醇、戊二醇、丙二醇、乙基己基甘油、聚乙二醇、山梨糖醇或它们的组合。在优选的实施方式中，多元醇是重量比为1-9:1-9的丁二醇和乙基己基甘油的混合物。
又一方面，本发明涉及中药提取物的冻干产品，所述冻干产品是无定形粉末。
又一方面，本发明涉及所述中药提取物、所述组合物、或所述冻干产品在皮肤抗糖化中的应用。
又一方面，本发明涉及一种皮肤外用剂，所述皮肤外用剂包含所述中药提取物、所述组合物、或所述冻干产品。
附图说明
下面结合附图进一步说明本发明。
图1显示了实施例4中药提取物的PGG的HPLC图谱。
图2是免疫组化检测结果。采用倒置Olympus显微镜，型号CKX41，放大倍数为40x。
图3显示了经萃取、浓缩、纯化处理后的中药提取物与多元醇混合样品。从左到右分别为实施例43，实施例121，实施例126的图片。
具体实施方式
本发明目的基于中药古方复合物，在协同搭配提升功效的同时，经过萃取工艺提高活性成分含量，改善中药复合物的特殊气味和颜色，改善中药复合物的生物利用度，并与现代化妆品学技术相结合，提供一种具有较 好的抗肌肤糖化作用的中药复合产物，可直接用于皮肤，也可作为功效添加剂加入到化妆品配方中使用，大幅提升化妆品应用的功效，且成本较低。
本发明根据多部古书籍和现代中草药学的研究成果，对多个中药古方进行复合得到较优复合中医配伍理论的美容方剂。本发明采用茯苓、蒺藜、白蔹、芍药、白术、白芨和白鲜皮得到中药提取物。
本发明利用现代中草药萃取方法和工艺对中药古方进行前处理。在优选的实施方式中，本发明中药提取物在溶剂提取之前包括将中草药原料进行粉碎。例如，可以采用高速万能粉碎机(FW100型)粉碎中草药原料的混合物。
本发明还利用浓缩、纯化等工艺，改善和提升中药复合物提取物中活性物含量。在一些实施方式中，采用中药材提取浓缩罐(TNH型)实现浓缩。
本发明利用化妆品技术，进一步改善肌肤对于中草药活性成分的生物利用度，实现抑制肌肤糖基化反应终端产物AGEs的效果。
除非另有定义，否则本文使用的所有科学技术术语的含义与本发明所属领域的普通技术人员通常所理解的含义相同。另外，将本文提及的所有出版物、专利申请、专利及其他参考文献以引用方式并入本文中。除非另有指明，否则百分比是指重量百分比。
中草药药材
芍药为毛茛科植物芍药Paeonia lactiflora Pall的干燥根，含有根含芍药甙(Paeoniflorin)、牡丹酚(Paeonol)、芍药花甙(Paeonin)，苯甲酸、挥发油、脂肪油、树脂、鞣质、糖、淀粉、粘液质、蛋白质、β-谷甾醇和三萜类等。
白蔹为葡萄科植物白蔹Ampelopsis japonica(Thunb.)Makino的干燥块根，根块含粘质和淀粉。
蒺藜为蒺藜科植物蒺藜Tribulus terrestris L.的干燥成熟果实。根和叶含皂甙，甙元有薯蓣皂甙元(Diosgenin)、芰脱皂甙元(Gitogenin)、绿莲皂甙元(Chlorogenin)、罗斯考皂甙元(Ruscogenin)。叶尚含山柰酚(Kaempferol) 和多种山柰酚甙。
白术为菊科植物白术Atractylodes macrocephala Koldz.的干燥根茎，含挥发油1.4％，主要成分为苍术醇(Atractylol)、苍术酮(Atractylon)等，并含有维生素A。
茯苓为多孔菌科真菌茯苓Poria cocos(Schw.)Wolf的干燥菌核，含β-茯苓聚糖(β-Pachyman)约占干重93％和三萜类化合物乙酰茯苓酸(Pachymic acid)、茯苓酸(Tumulosic acid)、3β-羟基羊毛甾三烯酸[3β-Hydroxylanosta-7,9(11),24-trien-21-oil acid]。此外，尚含树胶、甲壳质、蛋白质、脂肪、甾醇、卵磷脂、葡萄糖、腺嘌呤、组氨酸、胆碱、β-茯苓聚糖分解酶、脂肪酶、蛋白酶等。
白鲜为芸香科植物白鲜Dictamnus dasycarpus Turcz.的干燥根皮，含白鲜碱(Dictamnine)、白鲜内酯(Dictamnolactone,Obaculactone,Limonin)、谷甾醇(Sitosterol)、黄柏酮酸(Obacunonic acid)、胡芦巴碱(Trigonelline)、胆碱(Choline)、梣皮酮(Fraxinellone)。尚含菜油甾醇(Campesterol)、茵芋碱(Skimmianin)、γ-崖椒碱(γ-Fagarin)、白鲜明碱(Dasycarpamin)。地上部分含有补骨脂素(Psoralen)和花椒毒素(Xanthotoxin)。
白芨为兰科植物白及Bletilla striafa(Thunb.)Reichb.f.的干燥块茎，其新鲜块茎含水分14.6％、淀粉30.48％，葡萄糖1.5％。又含挥发油、粘液质。根含白及甘露聚糖(Bletilla mannan)，是由4份甘露糖和1份葡萄糖组成的葡配甘露聚糖。
遵循中药美容古方配伍的组方思路，仍然以阴阳共养之物为底，即芍药(Paeonia lactiflora Pall)、白蔹(Ampelopsis japonica(Thunb Vlakino)为君药；臣药则分别配伍温煦之蒺藜(Tribulus terrestris L)、白术(Atractylodes macrocephala Koidz)中草药；另加佐药茯苓(Poria cocos(Schw.)Wolf)、白鲜(Dictamnus dasycarpus Turcz)；使药为白芨(Bletilla striata Thunb Reichb.f.)。参考2015版《中国药典》一部，与中国植物志等内容进行鉴定。组方尽量精简，组成平衡美肌之简方，以符合后期提取、纯化技术等工艺需求。
中药提取物
本申请首次将茯苓、蒺藜、白蔹、芍药、白术、白芨和白鲜皮组合进行提取物，得到的复方中药提取物中各味药之间相辅相成，以达到抗糖化、祛黄、美白、提亮肤色的功效。
在一些实施方式中，本发明的中药提取物中原料茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮的重量比为1:1-4:4-10:8-10:1-4:1-4:1-4。在一个具体的实施方式中，本发明的中药提取物中原料茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮的重量比为1:4:10:10:1:4:4。在一个具体的实施方式中，本发明的中药提取物中原料茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮的重量比为1:2:8:10:4:1:2。在一个具体的实施方式中，本发明的中药提取物中原料茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮的重量比为1:1:10:10:4:1:1。在一个具体的实施方式中，本发明的中药提取物中原料茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮的重量比为1:1:4:10:4:1:1。在一个具体的实施方式中，本发明的中药提取物中原料茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮的重量比为1:1:4:10:2:1:1。在一个具体的实施方式中，本发明的中药提取物中原料茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮的重量比为1:2:4:8:4:1:2。
本发明的中药提取物可以作为功效添加剂加入到化妆品中使用，以达到抗糖化、祛黄、美白、提亮肤色的功效。
提取方法
本发明的中药提取物通过溶剂提取法制备。常见提取溶剂包括但不限于：水(例如去离子水)、有机溶剂(例如乙醇)。还有其他的提取方式，包括微波提取、超临界二氧化碳提取、超声波提取、酶提取法等。
在一些实施方式中，本发明的中药提取物采用水提醇沉进行制备。例如，本发明的中药提取物采用去离子水煮沸提取，然后进行醇沉，获取滤液，得到所需的中药提取物。在一个具体的实施方式中，醇沉过程采用乙醇。在一些实施方式，水提醇沉的提取过程持续1-24小时。在一些实施方式，水提醇沉的提取过程持续12-16小时。
在一些实施方式中，本发明的中药提取物采用醇作为提取溶剂。例如，本发明的中药提取物可以采用乙醇提取。在优选的实施方式中，提取过程在煮沸条件下(例如80±10℃)进行。在优选的实施方式中，可以采用70体积％的乙醇进行提取。在一些实施方式中，醇提取过程持续2-4小时。
中药提取物与多元醇的组合物
本发明还涉及包含的中药提取物和多元醇的组合物。
本发明所用术语“多元醇”是指分子中具有2个及以上羟基的水溶性多元醇，包括但不限于：甘油、双甘油、丁二醇、戊二醇、丙二醇、乙基己基甘油、聚乙二醇、山梨糖醇等。多元醇在化妆品中常用作保湿剂，从环境中吸收水分，补充到肌肤中，增加皮肤表面的含水量。多元醇可以单独使用，或者它们可以组合使用。例如，可以将2种或更多种多元醇组合使用。
在一些实施方式中，将本发明的中药提取物与重量比为9:1的戊二醇和丙二醇的混合物进行组合。在一些实施方式中，将本发明的中药提取物与重量比为5:5的聚乙二醇和辛甘醇的混合物进行组合。
从软化细胞膜层和角质层角度看，丁二醇，戊二醇，乙基己基甘油是优选的。因此，在优选的实施方式中，将中药提取物与重量比为9:1的丁二醇和乙基己基甘油的混合物进行组合。
冻干产品
本发明还涉及中药提取物的冻干产品。在一些实施方式中，冻干产品可以的无定形粉末。
在一些实施方式中，采用北京博医康实验仪器有限公司的真空冷冻干燥机FD-2型进行冻干。
在一些实施方式中，冷冻干燥设置-80～-40℃，真空度1～30Pa，时间8小时，升华干燥设置-20～+20℃，真空度1～30Pa，时间6小时，含水量控制在0～6％之间，解析干燥设置-10～+20℃，真空度1～30Pa，时间3小时，含水量控制在0～6％之间。
皮肤外用剂
本发明还涉及一种皮肤外用剂，该皮肤外用剂包含本发明的中药提取物、中药提取物与多元醇的组合物、或中药提取物的冻干产品，以及化妆品、皮肤病学或药学上可接受的赋形剂。
在一些优选的实施方式中，以所述皮肤外用剂的总重量计，中药提取物的用量为0.001-50％(w/w)，优选为1-20％(w/w)，更优选为2-10％(w/w)，最优选为3-8％(w/w)。
或者，中药提取物的冻干产品也可以单独使用。
所述皮肤外用剂组合物是通常用于皮肤外部的所有成分的统称概念，例如可以是化妆料组合物或药学组合物。所述化妆料组合物中可以是基础化妆料、面部妆容化妆料、身体用化妆料、头发护理用化妆料等，对其剂型无特殊限制，根据不同目的可合理选择。
这种皮肤外用剂可以配制成任何合适的产品形式。这类产品形式包括，但不限于气溶胶型喷雾剂、霜剂、乳液、固体、液体、分散体、泡沫、凝胶、化妆水、摩丝、软膏、粉剂、贴剂、润发油、溶液、手按泵型喷雾剂、棒状物、面膜和湿纸巾。可以这种皮肤外用剂通过本领域众所周知的各种方法便利地用于制备或作为化妆品、皮肤病学或药物局部施用产品。
本发明的皮肤外用剂组合物可以包括一种或多种下列成分：抗过敏剂、抗微生物剂、抗氧化剂、螯合剂、着色剂去色素剂、润肤剂、乳化剂、表皮脱落剂、成膜剂、香料、保湿剂、昆虫驱避剂、润滑剂、药物活性剂、增湿剂、耐光剂、防腐剂、护肤剂、皮肤渗透增强剂、防晒剂、稳定剂、表面活性剂、增稠剂、粘度调节剂、维生素或其任意组合。
可以用于本发明皮肤外用剂组合物的化妆品、皮肤病学或药学上可接受的赋形剂为水相、油相、凝胶、水包蜡型乳液、水包油型乳液或油包水型乳液的形式。水相为一种或多种水溶性或分散性组分的混合物，其在室温(25℃)下可以为液体、半固体或固体。赋形剂包括或可以为在水或水-醇赋形剂中的混悬液、分散液或溶液的形式，其可以含有增稠剂或凝胶剂。本领域技术人员可以基于本领域技术人员掌握的知识选择合适的产品形式，其中包含的组分。
所述的组合物可以包括水相，该水相可以含有水或水与至少一种亲水性有机溶剂的混合物，所述的亲水性有机溶剂诸如醇，尤其是含有2-5个碳原子的直链或支链低级一元醇，如乙醇或丙醇；多元醇，如丙二醇、山梨醇、甘油、泛醇或聚乙二醇及其混合物。
当发明的组合物为乳液形式时，该组合物还可以任选包含表面活性剂。
所述的组合物还可以包含成膜聚合物，如聚氨基甲酸酯、聚丙烯酸均聚物或共聚物、聚酯、基于烃的树脂和/或硅氧烷树脂。可以将聚合物溶于或分散于化妆品可接受的赋形剂中并且任选与增塑剂合并。
本发明的组合物还可以包含油相，所述的油相含有在室温(25℃)下为液体的油溶性或油分散性组分和/或在室温下为油状或蜡状的物质，如蜡、半固体、树胶及其混合物。该油相还可以含有有机溶剂。
通常在室温下为液体，合适的油性物质包括：来源于动物的基于烃的油，如全氢化角鲨烯；基于烃的植物油，如液体的C4-10脂肪酸的甘油三酯类，例如庚酸或辛酸甘油三酯类，或油，例如向日葵油、玉米油、大豆油、葡萄籽油、蓖麻油、鳄梨油、辛酸/癸酸甘油三酯类、霍霍巴油；矿物或合成来源的直链或支链烃类，例如液体石蜡及其衍生物、凡士林；合成酯类和醚类，特别是脂肪醇的酯类，例如肉豆蔻酸异丙酯、棕榈酸2-乙基己酯、硬脂酸2-辛基十二烷基酯、异硬脂酸异硬脂醇酯；羟基化酯类，例如乳酸异硬脂醇酯、羟基硬脂酸辛酯、羟基硬脂酸辛酯、羟基硬脂酸辛基十二烷基酯、脂肪醇的庚酸酯类、辛酸酯类和癸酸脂类；多元醇酯类，例如丙二醇二辛酸酯、新戊二醇二庚酸酯、二甘醇二异壬酸酯和季戊四醇酯类；含有C12-26的脂肪醇类，例如辛基十二烷醇、2-丁基辛醇、2-己基癸醇、2-十一烷基十五烷醇、油醇；基于部分烃的氟油和/或氟硅油，硅油，在室温下为液体或半固体的挥发性或非挥发性的直链或环状聚甲基硅氧烷，例如环状聚二甲基硅氧烷和聚二甲基硅氧烷，其任选包含苯基，例如苯基三甲基硅氧烷、硅氧烷及其混合物。
本发明的组合物可以进一步包含常用于化妆品领域中的任何组分。这些组分包括防腐剂、水相增稠剂(提取物生物聚合物、合成聚合物)和脂肪相增稠剂、芳香剂、亲水性和亲脂性活性剂及其混合物。
本发明的组合物还可以包含另外的颗粒相，所述的颗粒相可以为化妆品组合物中使用的颜料和/或珠光剂和/或填充剂。
颜料可以存在于组合物中，合适的无机颜料包括氧化钛、氧化锆和氧化铈以及氧化锌、氧化铁和铁蓝；合适的有机颜料包括钡、锶、钙和铝色淀和碳黑。
珠光剂可以存在于组合物中，合适的珠光剂包括涂覆了氧化钛、氧化铁或天然颜料的云母。
填充剂可以存在于组合物中，合适的填充剂包括滑石粉、二氧化硅、硬脂酸锌、云母、高岭土、尼龙粉末、聚乙烯粉末、特氟龙、淀粉、一氮化硼、共聚物微球，例如硅氧烷树脂微珠。
本发明组合物的油相可以包含一种或多种蜡、树胶或其混合物。蜡包括基于烃的蜡、氟蜡和/或硅氧烷蜡，并且可以来源于植物、矿物、动物和/或合成来源。合适的蜡包括蜂蜡、巴西棕榈蜡、小烛树蜡、石蜡、微晶蜡、地蜡；合成蜡包括聚乙烯蜡、含有C16-45的硅氧烷蜡。树胶一般为聚二甲基硅氧烷或羧甲基纤维素钠或提取物类，并且半固体物质一般为基于烃的化合物，如羊毛脂及其衍生物。
可以将本发明的组合物配制成任何合适的产品形式。这类产品形式包括，但不限于气溶胶型喷雾剂、霜剂、乳液、固体、液体、分散体、泡沫、凝胶、化妆水、摩丝、软膏、粉剂、贴剂、润发油、溶液、手按泵型喷雾剂、棒状物、面膜和湿纸巾。可以将本发明的组合物通过本领域众所周知的各种方法便利地用于制备或作为化妆品、皮肤病学或药物局部施用产品。
本发明的皮肤外用剂组合物可以包括一种或多种下列成分：抗过敏剂、抗微生物剂、抗氧化剂、螯合剂、着色剂去色素剂、润肤剂、乳化剂、表皮脱落剂、成膜剂、香料、保湿剂、昆虫驱避剂、润滑剂、药物活性剂、增湿剂、耐光剂、防腐剂、护肤剂、皮肤渗透增强剂、防晒剂、稳定剂、表面活性剂、增稠剂、粘度调节剂、维生素或其任意组合。
下面结合具体的实施例进一步阐述本发明。但是，应该明白，这些实施例仅用于说明本发明而不构成对本发明范围的限制。下列实施例中未注 明具体条件的试验方法，通常按照常规条件，或按照制造厂商所建议的条件。除非另有说明，所有的百分比和份数按重量计。
以下实施例中采用的药材原材料均购自“安徽亳州京皖中药饮片厂”，来料形态为“中草药饮片”；采用的生物试剂均购自上海家化生物科技有限公司。
以下实施例中采用的实验仪器如下：
1.固定搅拌器(IKA RW20)
2.恒温水浴锅上海一恒科学仪器有限公司HWS 28型
3.称量天平METTLER TOLEDO PL602-S
4.电热恒温CO2培养箱(Thermo,150i)
5.冰箱
6.Waters 2690HPLC，Waters 2487双波长吸收检测器
7.高速万能粉碎机(FW100型)
8.中药材提取浓缩罐(TNH型)
9.紫外可见分光光度仪(赛默飞)
10.真空冷冻干燥机，型号FD-2型，北京博医康实验仪器有限公司
11.超净工作台(苏州安泰，SW-CJ-IF)
12.酶标仪(BioTek,Epoch)
13.倒置显微镜(Olympus,CKX41)
14.正置显微镜(Olympus,BX53)
实施例1：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:4:10:10:1:4:4，混合后经高速万能粉碎机粉碎得粗粉1kg，加12kg去离子水于提取浓缩罐中煮沸，保持微沸1小时，过滤，滤渣复提一次，再过滤，合并两次滤液，浓缩至1.5kg，冷却，边搅拌边加入体积浓度70％的乙醇，静置12小时，过滤，滤液减压浓缩至1kg，即得中药提取物备用。
实施例2：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:2:8:10:4:1:2，混合后经高速万能粉碎机粉碎得粗粉1kg，加12kg去离子水于提取浓缩罐中煮沸，保持微沸1小时，过滤，滤渣复提一次，再过滤，合并两次滤液，浓缩至1.5kg，冷却，边搅拌边加入乙醇至乙醇体积浓度70％，静置12小时，过滤，滤液减压浓缩至1kg，即得中药提取物备用。
实施例3：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:1:10:10:4:1:1，混合后经高速万能粉碎机粉碎得粗粉1kg，加12kg去离子水于提取浓缩罐中煮沸，保持微沸1小时，过滤，滤渣复提一次，再过滤，合并两次滤液，浓缩至1.5kg，冷却，边搅拌边加入乙醇至乙醇体积浓度70％，静置12小时，过滤，滤液减压浓缩至1kg，即得中药提取物备用。
实施例4：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:1:4:10:4:1:1，混合后经高速万能粉碎机粉碎得粗粉1kg，加12kg去离子水于提取浓缩罐中煮沸，保持微沸1小时，过滤，滤渣复提一次，再过滤，合并两次滤液，浓缩至1.5kg，冷却，边搅拌边加入乙醇至乙醇体积浓度70％，静置12小时，过滤，滤液减压浓缩至1kg，即得中药提取物备用。
实施例5：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:1:4:10:2:1:1，混合后经高速万能粉碎机粉碎得粗粉1kg，加12kg去离子水于提取浓缩罐中煮沸，保持微沸1小时，过滤，滤渣复提一次，再过滤，合并两次滤液，浓缩至1.5kg，冷却，边搅拌边加入乙醇至乙醇体积浓度70％，静置12小时，过滤，滤液减压浓缩至1kg，即得中药提取物备用。
实施例6：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:2:4:8:4:1:2，混合后经高速万能粉碎机粉碎得粗粉1kg，加12kg去离子水于提取浓缩罐中煮沸，保持微沸1小时，过滤，滤渣复提一次，再过滤，合并两次滤液，浓缩至1.5kg，冷却，边搅拌边加入乙醇至乙醇体积浓度70％，静置12小时，过滤，滤液减压浓缩至1kg，即得中药提取物备用。
实施例7：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:4:10:10:1:4:4，混合后经高速万能粉碎机粉碎得粗粉1kg，加10kg乙醇溶液(70％体积浓度)于提取浓缩罐中煮沸，保持温度80℃持续2小时，过滤，滤渣复提一次，再过滤，合并两次滤液，滤液减压浓缩至1kg，即得中药提取物备用。
实施例8：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:2:8:10:4:1:2，混合后经高速万能粉碎机粉碎得粗粉1kg，加10kg乙醇溶液(70％体积浓度)于提取浓缩罐中煮沸，保持温度80℃持续2小时，过滤，滤渣复提一次，再过滤，合并两次滤液，滤液减压浓缩至1kg，即得中药提取物备用。
实施例9：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:1:10:10:4:1:1，混合后经高速万能粉碎机粉碎得粗粉1kg，加10kg乙醇溶液(70％体积浓度)于提取浓缩罐中煮沸，保持温度80℃持续2小时，过滤，滤渣复提一次，再过滤，合并两次滤液，滤液减压浓缩至1kg，即得中药提取物备用。
实施例10：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:1:4:10:4:1:1，混合后经高速万能粉碎机粉碎得粗粉1kg，加10kg乙醇溶液(70％体积浓度)于提取浓缩罐中煮沸，保持温度80℃持续2小时，过滤，滤渣复提一次，再过滤，合并两次滤液，滤液减压浓缩至1kg，即得中药提取物备用。
实施例11：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:1:4:10:2:1:1，混合后经高速万能粉碎机粉碎得粗粉1kg，加10kg乙醇溶液(70％体积浓度)于提取浓缩罐中煮沸，保持温度80℃持续2小时，过滤，滤渣复提一次，再过滤，合并两次滤液，滤液减压浓缩至1kg，即得中药提取物备用。
实施例12：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:2:4:8:4:1:2，混合后经高速万能粉碎机粉碎得粗粉1kg，加10kg乙醇溶液(70％体积浓度)于提取浓缩罐中煮沸，保持温度80℃持续2小时，过滤，滤渣复提一次，再过滤，合并两次滤液，滤液减压浓缩至1kg，即得中药提取物备用。
实施例13：中药提取物的制备(对比例)
称取茯苓经高速万能粉碎机粉碎得粗粉1kg，加12kg去离子水于提取浓缩罐中煮沸，保持微沸1小时，过滤，滤渣复提一次，再过滤，合并两次滤液，浓缩至1.5kg，冷却，边搅拌边加入乙醇至乙醇体积浓度70％，静置12小时，过滤，滤液减压浓缩至1kg，即得中药提取物备用。
实施例14：中药提取物的制备(对比例)
称取蒺藜经高速万能粉碎机粉碎得粗粉1kg，加12kg去离子水于提取 浓缩罐中煮沸，保持微沸1小时，过滤，滤渣复提一次，再过滤，合并两次滤液，浓缩至1.5kg，冷却，边搅拌边加入乙醇至乙醇体积浓度70％，静置12小时，过滤，滤液减压浓缩至1kg，即得中药提取物备用。
实施例15：中药提取物的制备(对比例)
称取白蔹经高速万能粉碎机粉碎得粗粉1kg，加12kg去离子水于提取浓缩罐中煮沸，保持微沸1小时，过滤，滤渣复提一次，再过滤，合并两次滤液，浓缩至1.5kg，冷却，边搅拌边加入乙醇至乙醇体积浓度70％，静置12小时，过滤，滤液减压浓缩至1kg，即得中药提取物备用。
实施例16：中药提取物的制备(对比例)
称取芍药经高速万能粉碎机粉碎得粗粉1kg，加12kg去离子水于提取浓缩罐中煮沸，保持微沸1小时，过滤，滤渣复提一次，再过滤，合并两次滤液，浓缩至1.5kg，冷却，边搅拌边加入乙醇至乙醇体积浓度70％，静置12小时，过滤，滤液减压浓缩至1kg，即得中药提取物备用。
实施例17：中药提取物的制备(对比例)
称取白术经高速万能粉碎机粉碎得粗粉1kg，加12kg去离子水于提取浓缩罐中煮沸，保持微沸1小时，过滤，滤渣复提一次，再过滤，合并两次滤液，浓缩至1.5kg，冷却，边搅拌边加入乙醇至乙醇体积浓度70％，静置12小时，过滤，滤液减压浓缩至1kg，即得中药提取物备用。
实施例18：中药提取物的制备(对比例)
称取白芨经高速万能粉碎机粉碎得粗粉1kg，加12kg去离子水于提取浓缩罐中煮沸，保持微沸1小时，过滤，滤渣复提一次，再过滤，合并两次滤液，浓缩至1.5kg，冷却，边搅拌边加入乙醇至乙醇体积浓度70％，静置12小时，过滤，滤液减压浓缩至1kg，即得中药提取物备用。
实施例19：中药提取物的制备(对比例)
称取白鲜皮经高速万能粉碎机粉碎得粗粉1kg，加12kg去离子水于提取浓缩罐中煮沸，保持微沸1小时，过滤，滤渣复提一次，再过滤，合并两次滤液，浓缩至1.5kg，冷却，边搅拌边加入乙醇至乙醇体积浓度70％，静置12小时，过滤，滤液减压浓缩至1kg，即得中药提取物备用。
实施例20：中药提取物的制备(对比例)
称取茯苓经高速万能粉碎机粉碎得粗粉1kg，加10kg乙醇溶液(70％体积浓度)于提取浓缩罐中煮沸，保持温度80℃持续2小时，过滤，滤渣复提一次，再过滤，合并两次滤液，滤液减压浓缩至1kg，即得中药提取物备用。
实施例21：中药提取物的制备(对比例)
称取蒺藜经高速万能粉碎机粉碎得粗粉1kg，加10kg乙醇溶液(70％体积浓度)于提取浓缩罐中煮沸，保持温度80℃持续2小时，过滤，滤渣复提一次，再过滤，合并两次滤液，滤液减压浓缩至1kg，即得中药提取物备用。
实施例22：中药提取物的制备(对比例)
称取白蔹经高速万能粉碎机粉碎得粗粉1kg，加10kg乙醇溶液(70％体积浓度)于提取浓缩罐中煮沸，保持温度80℃持续2小时，过滤，滤渣复提一次，再过滤，合并两次滤液，滤液减压浓缩至1kg，即得中药提取物备用。
实施例23：中药提取物的制备(对比例)
称取芍药经高速万能粉碎机粉碎得粗粉1kg，加10kg乙醇溶液(70％体积浓度)于提取浓缩罐中煮沸，保持温度80℃持续2小时，过滤，滤渣复提一次，再过滤，合并两次滤液，滤液减压浓缩至1kg，即得中药提取物备用。
实施例24：中药提取物的制备(对比例)
称取白术经高速万能粉碎机粉碎得粗粉1kg，加10kg乙醇溶液(70％体积浓度)于提取浓缩罐中煮沸，保持温度80℃持续2小时，过滤，滤渣复提一次，再过滤，合并两次滤液，滤液减压浓缩至1kg，即得中药提取物备用。
实施例25：中药提取物的制备(对比例)
称取白芨经高速万能粉碎机粉碎得粗粉1kg，加10kg乙醇溶液(70％体积浓度)于提取浓缩罐中煮沸，保持温度80℃持续2小时，过滤，滤渣复提一次，再过滤，合并两次滤液，滤液减压浓缩至1kg，即得中药提取物备用。
实施例26：中药提取物的制备(对比例)
称取白鲜皮经高速万能粉碎机粉碎得粗粉1kg，加10kg乙醇溶液(70％体积浓度)于提取浓缩罐中煮沸，保持温度80℃持续2小时，过滤，滤渣复提一次，再过滤，合并两次滤液，滤液减压浓缩至1kg，即得中药提取物备用。
实施例27：中药提取物的制备(对比例)
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:0:10:10:1:4:4，混合后经高速万能粉碎机粉碎得粗粉1kg，加12kg去离子水于提取浓缩罐中煮沸，保持微沸1小时，过滤，滤渣复提一次，再过滤，合并两次滤液，浓缩至1.5kg，冷却，边搅拌边加入乙醇至乙醇体积浓度70％，静置12小时，过滤，滤液减压浓缩至1kg，即得中药提取物备用。
实施例28：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:2:0:10:4:1:2，混合后经高速万能粉碎机粉碎得粗粉1kg，加12kg去离子 水于提取浓缩罐中煮沸，保持微沸1小时，过滤，滤渣复提一次，再过滤，合并两次滤液，浓缩至1.5kg，冷却，边搅拌边加入乙醇至乙醇体积浓度70％，静置12小时，过滤，滤液减压浓缩至1kg，即得中药提取物备用。
实施例29：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:1:10:0:4:1:1，混合后经高速万能粉碎机粉碎得粗粉1kg，加12kg去离子水于提取浓缩罐中煮沸，保持微沸1小时，过滤，滤渣复提一次，再过滤，合并两次滤液，浓缩至1.5kg，冷却，边搅拌边加入乙醇至乙醇体积浓度70％，静置12小时，过滤，滤液减压浓缩至1kg，即得中药提取物备用。
实施例30：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:1:4:10:0:1:1，混合后经高速万能粉碎机粉碎得粗粉1kg，加12kg去离子水于提取浓缩罐中煮沸，保持微沸1小时，过滤，滤渣复提一次，再过滤，合并两次滤液，浓缩至1.5kg，冷却，边搅拌边加入乙醇至乙醇体积浓度70％，静置12小时，过滤，滤液减压浓缩至1kg，即得中药提取物备用。
实施例31：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:1:4:10:2:0:1，混合后经高速万能粉碎机粉碎得粗粉1kg，加12kg去离子水于提取浓缩罐中煮沸，保持微沸1小时，过滤，滤渣复提一次，再过滤，合并两次滤液，浓缩至1.5kg，冷却，边搅拌边加入乙醇至乙醇体积浓度70％，静置12小时，过滤，滤液减压浓缩至1kg，即得中药提取物备用。
实施例32：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:2:4:8:4:1:0，混合后经高速万能粉碎机粉碎得粗粉1kg，加12kg去离子水于提取浓缩罐中煮沸，保持微沸1小时，过滤，滤渣复提一次，再过滤， 合并两次滤液，浓缩至1.5kg，冷却，边搅拌边加入乙醇至乙醇体积浓度70％，静置12小时，过滤，滤液减压浓缩至1kg，即得中药提取物备用。
实施例33：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝0:2:4:8:4:1:0，混合后经高速万能粉碎机粉碎得粗粉1kg，加12kg去离子水于提取浓缩罐中煮沸，保持微沸1小时，过滤，滤渣复提一次，再过滤，合并两次滤液，浓缩至1.5kg，冷却，边搅拌边加入乙醇至乙醇体积浓度70％，静置12小时，过滤，滤液减压浓缩至1kg，即得中药提取物备用。
实施例34：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:0:10:10:1:4:4，混合后经高速万能粉碎机粉碎得粗粉1kg，加10kg乙醇溶液(70％体积浓度)于提取浓缩罐中煮沸，保持温度80℃持续2小时，过滤，滤渣复提一次，再过滤，合并两次滤液，滤液减压浓缩至1kg，即得中药提取物备用。
实施例35：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:2:0:10:4:1:2，混合后经高速万能粉碎机粉碎得粗粉1kg，加10kg乙醇溶液(70％体积浓度)于提取浓缩罐中煮沸，保持温度80℃持续2小时，过滤，滤渣复提一次，再过滤，合并两次滤液，滤液减压浓缩至1kg，即得中药提取物备用。
实施例36：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:1:10:0:4:1:1，混合后经高速万能粉碎机粉碎得粗粉1kg，加10kg乙醇溶液(70％体积浓度)于提取浓缩罐中煮沸，保持温度80℃持续2小时，过滤，滤渣复提一次，再过滤，合并两次滤液，滤液减压浓缩至1kg，即得中 药提取物备用。
实施例37：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:0:4:10:0:1:1，混合后经高速万能粉碎机粉碎得粗粉1kg，加10kg乙醇溶液(70％体积浓度)于提取浓缩罐中煮沸，保持温度80℃持续2小时，过滤，滤渣复提一次，再过滤，合并两次滤液，滤液减压浓缩至1kg，即得中药提取物备用。
实施例38：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:0:4:10:2:0:1，混合后经高速万能粉碎机粉碎得粗粉1kg，加10kg乙醇溶液(70％体积浓度)于提取浓缩罐中煮沸，保持温度80℃持续2小时，过滤，滤渣复提一次，再过滤，合并两次滤液，滤液减压浓缩至1kg，即得中药提取物备用。
实施例39：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝1:2:0:8:4:1:0，混合后经高速万能粉碎机粉碎得粗粉1kg，加10kg乙醇溶液(70％体积浓度)于提取浓缩罐中煮沸，保持温度80℃持续2小时，过滤，滤渣复提一次，再过滤，合并两次滤液，滤液减压浓缩至1kg，即得中药提取物备用。
实施例40：中药提取物的制备
按重量比例称取茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮＝0:2:0:8:4:1:0，混合后经高速万能粉碎机粉碎得粗粉1kg，加10kg乙醇溶液(70％体积浓度)于提取浓缩罐中煮沸，保持温度80℃持续2小时，过滤，滤渣复提一次，再过滤，合并两次滤液，滤液减压浓缩至1kg，即得中药提取物备用。
测试例1：活性物含量检测
取实施例1-40中药提取物分别检测其中活性物含量。
药物活性成分的化学结构是药物经皮渗透的关键。脂溶性的药物易于透过角质层和皮肤屏障、细胞膜，但在活性表皮中由于水分含量相对较高从而扩散较慢。而水溶性成分药物通过角质层细胞膜及皮肤屏障的速度较慢，但在活性表皮中扩散却较快。一般在选用功能性药物活性成分中，注重成分的分子结构，尽可能选择用油/水分配适中的药物成分，或者水溶性和脂溶性成分配合使用，往往能得到较好的效果。
1,2,3,4,6-O-没食子酰葡萄糖(PGG)是一种多酚类化合物，由5个没食子酸与1个葡萄糖通过酯键结合而成，相对分子量940，具有较好的亲水和亲油性，具有较强的生理活性，如抗多种肿瘤、抗氧化、抗白血病、抗过敏、降血糖等功能，存在于多种中药材中。
高效液相色谱法测定：
色谱条件与***适用性试验采用C18通用型Eclipse XDB-C18(4.6x250mm,5μm)色谱柱；以乙腈-0.1％稀磷酸(25:75)为流动相；梯度洗脱后，检测波长为230nm；柱温为35℃；流速为0.8mL/min；进样量为10μL。理论板数按照1,2,3,4,6-O-没食子酰葡萄糖(PGG)峰值计算应不低于200000。
对照品溶液的制备，精密称取干燥至恒重的PGG对照品，加入甲醇制成每1mL含0.2mg的混合溶液，摇匀即得。
待测样品制备，取实施例1-40的中药提取物各1mL，精密称定，置5mL量瓶中，加甲醇稀释至刻度，摇匀即得。
测定，分别精密吸取对照品溶液和待测样品溶液各10μL，注入高效液相色谱仪，记录HPLC图谱，得到图1。
测定实施例1-40含量结果如下表1所示。
表1：实施例1-40中药提取物中PGG含量
Figure PCTCN2022133255-appb-000001
Figure PCTCN2022133255-appb-000002
结果显示，经过不同的提取工艺，实施例1～6中药提取物获得了较高的PGG获得量，尤其以实施例2为最，达到了0.7913mg/ml，相比较实施例6高出了96％。对于实施例7～12而言，醇提的工艺即便是同样的中草药的配伍比例，在活性成分PGG的含量上，整体偏低，尤其是同样配比的实施例8，相较于实施例2来说，含量约低13.7％；同样地，在实施例1～6的水提醇沉工艺中，即便是同样的比例和配伍，整体的PGG含量高于实施例7～12醇提工艺。
对比不同配比的提取物，尤其是删减7味中草药材中的1味或多味的情况下，水提醇沉或者醇提工艺制备获得的PGG含量普遍低于7味药材共 同存在的情况，如实施例14，甚至出现了未检出PGG的情况。将实施例2与实施例13～40中获得的PGG含量进行比较，基本在2-100倍之间；总体而言，通过不同比例配伍获得的7种不同的中药材，不同的配伍和比例之间有较大的活性物含量的差异，而且对于不同的提取工艺，也存在着活性物含量的较大差异。
实施例41～80：中药提取物的后处理
分别将实施例1～40中药提取物1kg，分散在10kg丁二醇：乙基己基甘油混合液体(重量比9:1)中，搅拌至均匀混合，备用。
实施例81～120：中药提取物的后处理
分别将实施例1～40提取物1kg，冷冻干燥设置-80～-40℃，真空度1～30Pa，时间8小时，升华干燥设置-20～+20℃，真空度1～30Pa，时间6小时，含水量控制在0～6％之间，解析干燥设置-10～+20℃，真空度1～30Pa，时间3小时，含水量控制在0～6％之间，得淡黄色至黄棕色疏松多孔、海绵状无定型粉末，备用。
测试例2：抗糖化功效(检测指标AGEs)对比
获取ExVivo组织(由广东博溪生物科技有限公司提供)，去除皮下脂肪组织是采用常用工具辊轴取皮刀，将离体皮肤组织块脂肪端削薄以达到目的。去除皮下脂肪后的皮肤组织块低温(-20℃)贮藏冻存备用。
在含有双抗的PBS溶液解冻。青霉素-链霉素(10,000U/ml)混合液又称为双抗，即含有10000单位/ml青霉素和10000μg/ml链霉素的溶液，生产商为赛默飞世尔科技(中国)有限公司，即配制100ml的PBS溶液加入双抗溶液1ml，目的是起到联合抗革兰氏阳性菌和革兰氏阴性菌的作用，防止皮肤组织块受到细菌污染。磷酸盐缓冲溶液(PBS)是一种平衡盐溶液，生产商为北京索莱宝科技有限公司，P1020-500ml 1xPBS缓冲液(pH7.2-7.4)。
取直径4～6mm的组织块，在液氮和37℃水浴锅中反复冻融5次。将 离体皮肤组织块置于24孔板中，加1ml含有双抗的PBS，CO 2孵育18±2h。
糖化反应按照以下步骤进行：
1)取新的24孔板，加入0.5mM浓度甲基乙二醛，1ml/孔；
2)按照10J/cm 2UVA辐照剂量进行辐照。UVA辐照采用的是UVA紫外线光疗仪，生产商上海SIGMA高技术有限公司，按照辐照剂量10J/cm 2UVA进行辐照，每次72小时；
3)辐照结束后，将孔板转移至45℃烘箱进行糖化反应72h；
4)72h之后，吸取并倒弃糖化反应工作液(糖化反应工作液为甲基乙二醛工作液，即按照1:13000的比例，用含双抗的PBS溶液稀释至0.5mM，甲基乙二醛生产商为Sigma,M0252型号)，加入新配制的糖化反应工作液，即3mM浓度的氨基胍硫酸盐作为阳性对照，3重量％实施例1～120获得的提取物、后处理物或后处理物的水溶液，重复步骤2)至4)；
5)UVA辐照进行3次，第4次后，清洗离体皮肤后转移至新的糖化反应工作液，重复步骤3)和4)，共计持续21天。
6)糖化反应结束后，将离体皮肤组织转移至1.5mL EP管中，用4％多聚甲醛固定，进行包埋，切片，AGEs免疫组化染色。
对于免疫组化(IHC)过程，主要分为：
—采用4％多聚甲醛对离体皮肤组织块进行固化24小时，多聚甲醛生产商为国药集团，AR级分析纯。
—将皮肤组织块进行梯度酒精脱水，75％酒精1h，85％酒精1h，95％酒精1h，95％酒精1h，无水乙醇I 1h，无水乙醇II 1h，二甲苯I 1h，二甲苯II 1h。以上试剂均为分析纯，使用前未进一步处理。
—采用蜡包埋机对脱水处理后的皮肤组织进行包埋，于-20℃进行冷却，待蜡凝固后取出在切片机上进行切片；包埋机生产商为徕卡HistoCore Arcadia包埋机，切片机为徕卡CM1520型切片机。
—将新鲜的切片置于玻片上，滴加少量PBS缓冲液以防止干化及蛋白交联。PBS缓冲液购自赛默飞世尔科技(中国)有限公司，型号eBioscience IHC溶液。
—采用SP试剂盒(福州迈新公司生产，型号E-EL-0067C)说明书进 行免疫组化染色检测皮肤组织中的AGE(DAB染色)，在倒置Olympus显微镜下对每张切片选择3个视野观察计算数据。
AGEs数据＝相对IOD值/Area平均值
使用GraphPad Prism作图，结果表示为Mean±SD。各组间比较采用t-test统计分析。
相对改善率(％)＝(阴性对照—实施例相对平均值)/阴性对照x 100％
表2：实施例1-120抗糖化功效(检测指标AGEs)结果
样品名称 相对平均值 SD 相对改善率/％
空白对照 1.00 0.10 -
阴性对照 1.70 0.03 -
阳性对照 1.31 0.03 22.94
实施例1 1.52 0.02 10.59
实施例2 1.59 0.04 6.47
实施例3 1.56 0.05 8.24
实施例4 1.56 0.05 8.24
实施例5 1.54 0.04 9.41
实施例6 1.57 0.05 7.65
实施例7 1.64 0.04 3.53
实施例8 1.62 0.06 4.71
实施例9 1.65 0.06 2.94
实施例10 1.61 0.05 5.29
实施例11 1.59 0.05 6.47
实施例12 1.60 0.04 5.88
实施例13 1.70 0.02 0.00
实施例14 1.72 0.01 -1.18
实施例15 1.74 0.01 -2.35
实施例16 1.69 0.01 0.59
实施例17 1.68 0.01 1.18
实施例18 1.66 0.01 2.35
实施例19 1.63 0.01 4.12
实施例20 1.65 0.03 2.94
实施例21 1.79 0.04 -5.29
实施例22 1.82 0.03 -7.06
实施例23 1.70 0.05 0.00
实施例24 1.68 0.07 1.18
实施例25 1.69 0.05 0.59
实施例26 1.69 0.06 0.59
实施例27 1.60 0.05 5.88
实施例28 1.57 0.07 7.65
实施例29 1.49 0.07 12.35
实施例30 1.57 0.04 7.65
实施例31 1.66 0.03 2.35
实施例32 1.72 0.03 -1.18
实施例33 1.69 0.04 0.59
实施例34 1.65 0.05 2.94
实施例35 1.63 0.05 4.12
实施例36 1.66 0.04 2.35
实施例37 1.68 0.04 1.18
实施例38 1.69 0.03 0.59
实施例39 1.72 0.02 -1.18
实施例40 1.68 0.02 1.18
实施例41 1.29 0.01 24.12
实施例42 1.27 0.01 25.29
实施例43 1.21 0.01 28.82
实施例44 1.37 0.01 19.41
实施例45 1.30 0.01 23.53
实施例46 1.39 0.03 18.24
实施例47 1.37 0.04 19.41
实施例48 1.41 0.03 17.06
实施例49 1.45 0.05 14.71
实施例50 1.58 0.07 7.06
实施例51 1.40 0.05 17.65
实施例52 1.57 0.06 7.65
实施例53 1.53 0.05 10.00
实施例54 1.52 0.07 10.59
实施例55 1.55 0.07 8.82
实施例56 1.51 0.04 11.18
实施例57 1.53 0.03 10.00
实施例58 1.60 0.03 5.88
实施例59 1.61 0.04 5.29
实施例60 1.62 0.05 4.71
实施例61 1.66 0.05 2.35
实施例62 1.53 0.04 10.00
实施例63 1.65 0.04 2.94
实施例64 1.59 0.01 6.47
实施例65 1.54 0.01 9.41
实施例66 1.52 0.01 10.59
实施例67 1.42 0.01 16.47
实施例68 1.46 0.01 14.12
实施例69 1.47 0.01 13.53
实施例70 1.48 0.03 12.94
实施例71 1.54 0.04 9.41
实施例72 1.57 0.03 7.65
实施例73 1.52 0.05 10.59
实施例74 1.59 0.07 6.47
实施例75 1.55 0.05 8.82
实施例76 1.59 0.06 6.47
实施例77 1.63 0.05 4.12
实施例78 1.72 0.07 -1.18
实施例79 1.69 0.07 0.59
实施例80 1.51 0.04 11.18
实施例81 1.49 0.03 12.35
实施例82 1.56 0.03 8.24
实施例83 1.55 0.04 8.82
实施例84 1.43 0.05 15.88
实施例85 1.39 0.05 18.24
实施例86 1.48 0.04 12.94
实施例87 1.55 0.04 8.82
实施例88 1.48 0.01 12.94
实施例89 1.57 0.01 7.65
实施例90 1.64 0.01 3.53
实施例91 1.62 0.01 4.71
实施例92 1.65 0.01 2.94
实施例93 1.66 0.01 2.35
实施例94 1.69 0.03 0.59
实施例95 1.70 0.04 0.00
实施例96 1.72 0.03 -1.18
实施例97 1.70 0.05 0.00
实施例98 1.72 0.07 -1.18
实施例99 1.58 0.05 7.06
实施例100 1.63 0.06 4.12
实施例101 1.45 0.03 14.71
实施例102 1.43 0.04 15.88
实施例103 1.56 0.03 8.24
实施例104 1.47 0.05 13.53
实施例105 1.57 0.07 7.65
实施例106 1.63 0.05 4.12
实施例107 1.62 0.06 4.71
实施例108 1.65 0.05 2.94
实施例109 1.69 0.07 0.59
实施例110 1.62 0.07 4.71
实施例111 1.47 0.04 13.53
实施例112 1.49 0.03 12.35
实施例113 1.53 0.03 10.00
实施例114 1.57 0.04 7.65
实施例115 1.54 0.05 9.41
实施例116 1.48 0.03 12.94
实施例117 1.43 0.04 15.88
实施例118 1.47 0.02 13.53
实施例119 1.40 0.03 17.65
实施例120 1.51 0.02 11.18
中药化妆品的核心功效的发挥首先需要研究的就是活性成分的透皮吸收和利用的问题。当皮肤角质层中含水量低于10％后，角朊细胞排列状态更加紧密，减少了皮肤有效透入面积和物质透皮的空间，从而降低了药物的透皮吸收程度和效率。在这种皮肤环境下，即使加大药物用量，效果并无太大改善，所以功能性化妆品，首先要做的是把皮肤的含水量恢复到正常水平或处于水润状态，角质层含水10％或以上，表皮中其他各层10％～47％，当表皮中含水量达到10％～50％时，使得紧密的角朊细胞形成多孔和饱满状态，增加了皮肤的有效面积和接触的横截面，药物的透皮吸收速度大大提高，在皮肤保湿的环境下药物会发挥更好的作用。给皮肤补充水分及锁住水分的有效方法有很多，比如在配方中添加天然保湿剂(NMF)、透明质酸、卵磷脂、神经酰胺和胆固醇、多元醇等。但神经酰胺和胆固醇 对细胞的屏蔽作用较大，从而影响药物的吸收与利用；透明质酸(HA)由于分子量较大，其分子筛效应不利于药物活性成分的渗透和吸收利用，特别是配方体系pH≤5时，会增强透明质酸分子的网状交联，增加药物扩散的阻力。肌肤中糖化反应的发生在表皮层的深处及真皮层，任何功效活性物质如果无法通过皮肤屏障是无法到达糖化部位而起到实际的功效发挥。本申请的提取物能够实现透皮吸收，帮助美白物质渗透被肌肤细胞利用而达到抗糖化的效果。
对比了相同浓度下的提取物及其不同后处理方式和工艺产物对抑制AGEs的效果差异。
从实施例1～40的结果来看(见表2)，提取物绝大部分具有抗糖化的功效，从相对改善率维度来看，7种中草药组合经过水提醇沉提取工艺获得的提取物整体改善在6.5％以上，相比较醇提工艺来说，后者平均的改善率在3～7％之间，说明该制备工艺获得的提取物整体功效略弱，这与相同实施例提取物中PGG的含量也是有正相关性的；水提醇沉工艺获得的实施例1～6中，以实施例1改善率达到10.59％为最高，而其次为实施例5的改善率为9.41％。
对于后继删减1味中草药或者多味中草药的实施例，即便采用相同的水提醇沉或者醇提的制备工艺，其抗糖化的改善率也是大幅度地降低，甚至出现了负值，如实施例21，实施例22，实施例39；但也有例外的情况，即实施例29，抗糖化改善率达到了12.35％，为较高的改善性能。但是单味中草药的提取液，不论是水提醇沉还是醇沉制备工艺，获得的提取物的抗糖化改善率均不高。
从实施例41～46为实施例1～6提取物的后处理产物，通过与特定的多元醇类组合复配，较大幅度地提升了实施例1～6提取物通过水提醇沉工艺制得的提取物在抑制AGEs方面的改善率，比如实施例43提升到了28.82％，实施例42提升到了25.29％，实施例41提升到了24.12％，实施例44提升到了19.41％，平均的提升比例超过了10％，说明采用选定的两种醇类物质进行复配组合对后处理能较大程度提高了中药提取物的抗糖化功效。
对比两种多元醇同比例下的自身的抗糖化能力，基本来说没有任何的抗糖化功效，说明多元醇组合对中草药复合物的活性物在抗糖化功效的发挥具有较大的促进作用。
与此同时，与实施例1～6提取物后处理产物与前述较高含量的PGG有一定的相关性，多元醇的加入可以极大地提升中草药活性成分被细胞的利用率是可能的工作机制。
从实施例47～52为实施例7～12提取物的后处理工艺产物，同样通过与特定的多元醇组合复配，也较大幅度地提升了实施例7～12提取物通过醇提工艺制得的提取物在抑制AGEs方面的改善率，比如实施例47也提升到了19.41％，实施例48提升到了17.06％，实施例51提升到了17.65％，较实施例7～12提取物提升超过了10％。说明在醇提工艺下，通过加入丁二醇及乙基己基甘油可以较大幅度地提升活性物的抗糖化功效。
从实施例53～80，对比考察了删减1味或者多味中草药材的组合物之间，水提醇沉工艺制备提取物采用丁二醇和乙基己基甘油复配后处理获得的产物在抑制AGEs方面的效果，从提取物本身的较低的抗糖化抑制效果来看，通过多元醇处理后抑制的效果虽然后提升，但是整体提升的比例不大。说明只有7味中草药按照特定的配伍比例组合后，才能发挥较好的抑制糖化反应的效果，另外在7味中草药按照特定的配伍比例组合后，采用丁二醇和乙基己基甘油才能获得更好的抑制糖基化反应的效率提升，因为中药配伍理论始载于一千九百多年前的本草鼻祖——《神农本草经》：“有单行者，有相须者，有相使者，有相畏者，有相恶者，有相反者，有相杀者，凡此七情，合和时视之”。简言之，本发明中的中药组合出现了类似的协同作用。
中草药复方添加剂里面的主要功效物质为黄酮、多酚、多糖等，具体的组分的分布和含量多数依赖于萃取方法和手段，而且因为多数物质不易保存，储存不当易发生氧化、分解、挥发、变质等问题而导致功效物质的损失。制备工艺后期采用了先进的冻干技术，使得提取物中的活性成分能够较好的保留，可以在更大程度上保留活性成分的原汁原味，特别是热敏性的活性成分，另外，因在真空无氧下操作，因此，一些易氧化的活性成 分(如油脂类)也可以得到保护。在真空和低温下操作，微生物的生长和酶作用受到抑制，常温下能长久储存，而且不需添加任何防腐剂，减少微生物的侵蚀,延长产品的保质期。通过冻干技术处理实施例1～40获得了实施例81～120的冻干后处理产物，最大化地避免活性成分由于各种可能的原因储存不当而导致的活性物质的损失，从而影响抗击糖化反应的效果。
对冻干处理产物，即实施例81～120的抑制AGEs进行了抑制糖基化AGEs的实验，从表2的结果改善率来看，总体来说与提取物本身的测试结果相当，个别的实施例的测试结果有升高，如实施例85提升对AGEs的改善率到18.24％，实施例102提升对AGEs的改善率到15.88％，呈现了一定程度的增长。说明从提取物到经冻干工艺处理后，在整个提取物的储存过程中能够较好地保留提取物本身的活性和抑制糖基化AGEs的效果。
图2显示了实施例42和实施例43的实验样品平行三个相同实验的结果。与空白对照相比较，阴性对照呈现出-70％的负向改善，从图中看出糖基化反应程度及颜色呈现了较黯沉的颜色。阳性对照相较于阴性对照改善22.94％，但是对于实验样品实施例42，实施例43，图片颜色呈现明显的浅色，说明该两例实施例的抑制AGEs的效果较好，分别达到了25.29％和28.82％。
综上所述，按照中医智慧思想配伍的实施例1～5获得的提取物经过特定多元醇组合配比后得到的实施例41～45具有更好的抑制肌肤糖基化反应终端产物AGEs的效果，在此基础上，对比不同的多元醇组合物及其不同的比例情况下，再次进行抑制糖基化反应终端产物AGEs的对比。
实施例121～125：中药提取物的后处理(对比例)
分别将实施例1～5提取物1kg，分散在10kg戊二醇：丙二醇混合液体(重量比9:1)中，搅拌至均匀混合，备用。
实施例126～130：中药提取物的后处理(对比例)
分别将实施例1～5中药提取物1kg分散在10kg聚乙二醇：辛甘醇(重量比5:5)中，搅拌至均匀混合，备用。
实施例131：多元醇组合配比(对比例)
按照重量比9:1配制丁二醇：乙基己基甘油混合液体，搅拌均匀混合，备用。
实施例132：多元醇组合配比(对比例)
按照重量比9:1配制戊二醇：丙二醇混合液体，搅拌均匀混合，备用。
实施例133：多元醇组合配比(对比例)
按照重量比5:5配制聚乙二醇：辛甘醇混合液体，搅拌均匀混合，备用。
测试例3：抗糖化功效(检测指标AGEs)对比
获取ExVivo组织(由广东博溪生物科技有限公司提供)，去除皮下脂肪组织是采用常用工具辊轴取皮刀，将离体皮肤组织块脂肪端削薄以达到目的。去除皮下脂肪后的皮肤组织块低温(-20℃)贮藏冻存备用。
在含有双抗的PBS溶液解冻。青霉素-链霉素(10,000U/ml)混合液又称为双抗，即含有10000单位/ml青霉素和10000μg/ml链霉素的溶液，生产商为赛默飞世尔科技(中国)有限公司，即配制100ml的PBS溶液加入双抗溶液1ml，目的是起到联合抗革兰氏阳性菌和革兰氏阴性菌的作用，防止皮肤组织块受到细菌污染。磷酸盐缓冲溶液(PBS)是一种平衡盐溶液，生产商为北京索莱宝科技有限公司，P1020-500ml 1xPBS缓冲液(pH7.2-7.4)。
取直径4～6mm的组织块，在液氮和37℃水浴锅中反复冻融5次。将离体皮肤组织块置于24孔板中，加1ml含有双抗的PBS，CO 2孵育18±2h。
糖化反应按照以下步骤进行：
1)取新的24孔板，加入0.5mM浓度甲基乙二醛，1ml/孔；
2)按照10J/cm 2UVA辐照剂量进行辐照。UVA辐照采用的是UVA紫外线光疗仪，生产商上海SIGMA高技术有限公司，按照辐照剂量10J/cm 2 UVA进行辐照，每次72小时；
3)辐照结束后，将孔板转移至45℃烘箱进行糖化反应72h；
4)72h之后，吸取并倒弃糖化反应工作液(糖化反应工作液为甲基乙二醛工作液，即按照1:13000的比例，用含双抗的PBS溶液稀释至0.5mM，甲基乙二醛生产商为Sigma,M0252型号)，加入新配置的糖化反应工作液，即3mM浓度的氨基胍硫酸盐作为阳性对照，3重量％实施例121～133获得的提取物或者后处理物，重复步骤2)至4)；
5)UVA辐照进行3次，第4次后，清洗离体皮肤后转移至新的糖化反应工作液，重复步骤3)和4)，共计持续21天。
6)糖化反应结束后，将离体皮肤组织转移至1.5mL EP管中，用4％多聚甲醛固定，进行包埋，切片，AGEs免疫组化染色。
对于免疫组化(IHC)过程，主要分为：
—采用4％多聚甲醛对离体皮肤组织块进行固化24小时，多聚甲醛生产商为国药集团，AR级分析纯。
—将皮肤组织块进行梯度酒精脱水，75％酒精1h，85％酒精1h，95％酒精1h，95％酒精1h，无水乙醇I 1h，无水乙醇II 1h，二甲苯I 1h，二甲苯II 1h。以上试剂均为分析纯，使用前未进一步处理。
—采用蜡包埋机对脱水处理后的皮肤组织进行包埋，于-20℃进行冷却，待蜡凝固后取出在切片机上进行切片；包埋机生产商为徕卡HistoCore Arcadia包埋机，切片机为徕卡CM1520型切片机。
—将新鲜的切片置于玻片上，滴加少量PBS缓冲液以防止干化及蛋白交联。PBS缓冲液购自赛默飞世尔科技(中国)有限公司，型号eBioscience IHC溶液。
—采用SP试剂盒(福州迈新公司生产，型号E-EL-0067C)说明书进行免疫组化染色检测皮肤组织中的AGE(DAB染色)，在倒置Olympus显微镜下对每张切片选择3个视野观察计算数据。
AGEs数据＝相对IOD值/Area平均值
使用GraphPad Prism作图，结果表示为Mean±SD。各组间比较采用t-test统计分析。
相对改善率(％)＝(阴性对照—实施例相对平均值)/阴性对照x 100％。
表3：实施例121～133抗糖化功效(检测指标AGEs)结果
样品名称 相对平均值 SD 相对改善率/％
空白对照 1.00 0.10 -
阴性对照 1.70 0.03 -
阳性对照 1.31 0.03 22.94
实施例121 1.36 0.05 20.00
实施例122 1.41 0.05 17.06
实施例123 1.38 0.04 18.82
实施例124 1.31 0.05 22.94
实施例125 1.39 0.04 18.24
实施例126 1.41 0.06 17.06
实施例127 1.40 0.06 17.65
实施例128 1.41 0.01 17.06
实施例129 1.47 0.01 13.53
实施例130 1.40 0.01 17.65
实施例131 1.69 0.03 0.59
实施例132 1.72 0.02 -1.18
实施例133 1.67 0.04 1.76
表4：实施例1～5提取物及不同的后处理产物抑制AGEs的改善率比较
Figure PCTCN2022133255-appb-000003
Figure PCTCN2022133255-appb-000004
对比相同浓度下的实施例1～5中药提取物经过不同处理产物对抑制AGEs的效果差异，其中实施例121～125为戊二醇+丙二醇的后处理产物，实施例126～130为聚乙二醇+辛甘醇的后处理产物，与水提醇沉制备工艺获得的提取物在抑制糖基化反应终端产物AGEs上面的横向对比见表4。从表中可以看出，采用丁二醇+乙基己基甘油的组合对实施例1～5的后处理获得的产物对糖基化反应终端产物AGEs的改善率最高。
采用戊二醇+丙二醇，及聚乙二醇+辛甘醇组合一定程度提高了中药提取物的抗糖化功效，说明不同的多元醇组合也对中草药复合物的活性物在抗糖化功效的发挥具有促进作用。与前述较高含量的PGG有一定的相关性的同时，不同种类多元醇的加入也引起了中草药活性成分被细胞的利用率的差异。
皮肤外用剂优选为化妆品组合物，例如化妆水、精华液、乳霜等。所述复合提取物在皮肤外用剂中的重量百分比为0.001％-50％(w/w)。优选的重量百分比为1％-20％(w/w)。更优选的重量百分比为2％-10％(w/w)。最优选的重量百分比为3％-8％(w/w)。
以下是中药提取物在皮肤外用剂中的具体应用的实施例，及其这些剂型的配方和制备方法。以下各表中“-”表示无添加。
应用例1：面霜的制备
Figure PCTCN2022133255-appb-000005
Figure PCTCN2022133255-appb-000006
应用例2：乳液的制备
Figure PCTCN2022133255-appb-000007
Figure PCTCN2022133255-appb-000008
应用例3：啫喱的制备
Figure PCTCN2022133255-appb-000009
应用例4：化妆水的制备
Figure PCTCN2022133255-appb-000010
Figure PCTCN2022133255-appb-000011
应用例5：精华液的制备
Figure PCTCN2022133255-appb-000012
应用例6：面膜的制备
Figure PCTCN2022133255-appb-000013
应用例7：眼霜的制备
Figure PCTCN2022133255-appb-000014
Figure PCTCN2022133255-appb-000015
应用例8：沐浴露的制备
Figure PCTCN2022133255-appb-000016
应用例9：洗面奶的制备
Figure PCTCN2022133255-appb-000017
本发明中的实施例82～121也可以作为低温冷冻干燥产品单独使用。
利用本发明的组合物制得的上述化妆品取得与市售的产品相当或更好的使用效果。
本发明通过上述实施例来说明本发明的详细方法，但本发明并不局限于上述详细方法，即不意味着本发明必须依赖上述详细方法才能实施。所属技术领域的技术人员应该明了，对本发明的任何改进，对本发明产品各原料的等效替换及辅助成分的添加、具体方式的选择等，均落在本发明的保护范围和公开范围之内。

Claims (12)

  1. 一种中药提取物，通过以下原料制备得到：茯苓、蒺藜、白蔹、芍药、白术、白芨和白鲜皮，其中茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮的重量比为1:1-4:4-10:8-10:1-4:1-4:1-4。
  2. 如权利要求1所述的中药提取物，所述提取物具有抗糖化作用。
  3. 对如权利要求1或2所述的中药提取物，所述提取物通过溶剂提取制备得到。
  4. 如权利要求3所述的中药提取物，其中，所述溶剂提取采用水提醇沉方法。
  5. 如权利要求3所述的中药提取物，其中，所述溶剂提取采用醇溶剂进行提取。
  6. 如权利要求4-5中任一项所述的中药提取物，其中，茯苓：蒺藜：白蔹：芍药：白术：白芨：白鲜皮的重量比为1:1-2:4-8:8-10:1-4:1-4:1-2。
  7. 一种组合物，所述组合物包含如权利要求4-5所述的中药提取物以及多元醇。
  8. 如权利要求7所述的组合物，其中，所述多元醇选自：甘油、双甘油、丁二醇、戊二醇、丙二醇、乙基己基甘油、聚乙二醇、山梨糖醇或它们的组合。
  9. 如权利要求7所述的组合物，其中，所述多元醇是重量比为1-9:1-9的丁二醇和乙基己基甘油的混合物。
  10. 如权利要求1-6中任一项所述的中药提取物的冻干产品，所述冻干产品是无定形粉末。
  11. 如权利要求1-6中任一项所述的中药提取物、如权利要求7-9中任一项所述的组合物、或如权利要求10所述的冻干产品在皮肤抗糖化中的应用。
  12. 一种皮肤外用剂，所述皮肤外用剂包含如权利要求1-6中任一项所述的中药提取物、如权利要求7-9中任一项所述的组合物、或如权利要求10所述的冻干产品。
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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106309911A (zh) * 2016-08-19 2017-01-11 上海家化联合股份有限公司 抗衰老中药组合物及其制备方法
CN110448505A (zh) * 2019-09-02 2019-11-15 重庆市药物种植研究所 一种可撕拉中药粉末面膜的制备方法及其产品
CN110664677A (zh) * 2019-09-27 2020-01-10 王英 一种具有抗糖化与抗皮肤衰老的组合物制备方法
CN113332216A (zh) * 2021-03-26 2021-09-03 广州市焱炎生物科技有限公司 一种面膜套组及其制备方法和使用方法
CN114392323A (zh) * 2021-12-30 2022-04-26 上海家化联合股份有限公司 抗糖化的中药提取物

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111214618A (zh) * 2020-03-06 2020-06-02 于忠 一种祛斑中药组合物及其应用
CN112641856B (zh) * 2020-12-29 2022-03-25 上海家化联合股份有限公司 抗衰老的中药组合物
CN113116750A (zh) * 2021-04-29 2021-07-16 广州睿森生物科技有限公司 一种多元醇组合物及其在化妆品中的应用

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106309911A (zh) * 2016-08-19 2017-01-11 上海家化联合股份有限公司 抗衰老中药组合物及其制备方法
CN110448505A (zh) * 2019-09-02 2019-11-15 重庆市药物种植研究所 一种可撕拉中药粉末面膜的制备方法及其产品
CN110664677A (zh) * 2019-09-27 2020-01-10 王英 一种具有抗糖化与抗皮肤衰老的组合物制备方法
CN113332216A (zh) * 2021-03-26 2021-09-03 广州市焱炎生物科技有限公司 一种面膜套组及其制备方法和使用方法
CN114392323A (zh) * 2021-12-30 2022-04-26 上海家化联合股份有限公司 抗糖化的中药提取物

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
SHANGHAI JAHWA UNITED CO., LTD.: "Herborist Advanced Brightening & Revitalizing Mask", DOMESTIC GENERAL COSMETIC RECORD INFORMATION, XP009547584, Retrieved from the Internet <URL:https://hzpba.nmpa.gov.cn/gccx/gxxcGC.html?prodId=1037803620550049792&gb=G> *
SHANGHAI JAHWA UNITED CO., LTD.: "Herborist Advanced Whitening Revitalizing Toner", DOMESTIC GENERAL COSMETIC RECORD INFORMATION, XP009547586, Retrieved from the Internet <URL:https://hzpba.nmpa.gov.cn/gccx/chakanHis.html?prodId=20171225230054968f2&gb=G> *
SHIXIA GUAN, CAO LIPING, LI HAIGANG, LI ZHONGGUI, XU SHUQIN, LI DUJUN, ZHOU YUBIN, YUAN ZHONGWEN: "Preparation and Quality Control of Freckle Removing Ointment", HERALD OF MEDICINE, vol. 30, no. 7, 1 July 2011 (2011-07-01), pages 941 - 943, XP093075079 *

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