WO2018045943A1 - 茯苓萃取物及其活性成分于保养肌肤及/或促进伤口愈合的用途 - Google Patents

茯苓萃取物及其活性成分于保养肌肤及/或促进伤口愈合的用途 Download PDF

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Publication number
WO2018045943A1
WO2018045943A1 PCT/CN2017/100526 CN2017100526W WO2018045943A1 WO 2018045943 A1 WO2018045943 A1 WO 2018045943A1 CN 2017100526 W CN2017100526 W CN 2017100526W WO 2018045943 A1 WO2018045943 A1 WO 2018045943A1
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Prior art keywords
acid
skin
extract
use according
preparation
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PCT/CN2017/100526
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English (en)
French (fr)
Inventor
林汉钦
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杏辉天力（杭州）药业有限公司
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Priority to JP2019533271A priority Critical patent/JP6798030B2/ja
Priority to KR1020197009564A priority patent/KR102295786B1/ko
Priority to MYPI2019001172A priority patent/MY195806A/en
Publication of WO2018045943A1 publication Critical patent/WO2018045943A1/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
    • A61K8/9728—Fungi, e.g. yeasts
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/194—Carboxylic acids, e.g. valproic acid having two or more carboxyl groups, e.g. succinic, maleic or phthalic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/575—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of three or more carbon atoms, e.g. cholane, cholestane, ergosterol, sitosterol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/726—Glycosaminoglycans, i.e. mucopolysaccharides
    • A61K31/728—Hyaluronic acid
    • 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
    • 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
    • A61K8/63—Steroids; Derivatives thereof
    • 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
    • A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
    • 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
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • 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
    • A61P17/16—Emollients or protectives, e.g. against radiation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • 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
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/318—Foods, ingredients or supplements having a functional effect on health having an effect on skin health and hair or coat
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2250/00—Food ingredients
    • A23V2250/20—Natural extracts
    • A23V2250/208—Fungi extracts
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • 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
    • A61K2800/52—Stabilizers
    • A61K2800/522—Antioxidants; Radical scavengers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/365—Lactones
    • A61K31/375—Ascorbic acid, i.e. vitamin C; Salts thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca

Definitions

  • the invention relates to the use of sputum extracts and/or active ingredients thereof, and in particular to the use of sputum extracts and/or active ingredients thereof to maintain skin and/or promote wound healing.
  • the cockroach extract and/or its active ingredient maintains the skin and/or promotes wound healing by enhancing the expression of collagen and/or hyaluronic acid.
  • the keratinocytes on the surface of the skin will accumulate due to the inability to fall off properly, which may cause the skin to lose its luster and elasticity, appear dry, rough, and even wrinkle and other skin aging phenomena. Therefore, if it can effectively increase the moisture content of the skin, it will reduce the occurrence of the above situation.
  • collagen and hyaluronic acid have the function of increasing the moisture content of the skin. Therefore, many skin care products and health products containing collagen and/or hyaluronic acid are claimed to provide moisture. Skin, firm skin, repair skin, moisturize, diminish wrinkles, delay skin aging, and prevent skin aging. In addition, since collagen is highly biocompatible, low in immunity, and can be decomposed and absorbed by the body, there are also health foods which are claimed to provide effects for promoting wound healing due to the addition of collagen.
  • the inventors of the present invention have found that the extract of sputum and/or its active ingredient can effectively increase the content of collagen, extracellular collagen, and/or extracellular hyaluronic acid in the cells, that is, sputum extract and/or its activity.
  • the ingredients are effective in enhancing the content of endogenous collagen and/or hyaluronic acid, so they can be used to maintain the skin and/or promote wound healing.
  • the cockroach extract comprises at least one of: dehydropachymic acid (DPA), pachymic acid (PA), dehydrotumulosic acid (DTA), and tolunic acid.
  • DPA dehydropachymic acid
  • PA pachymic acid
  • DTA dehydrotumulosic acid
  • tolunic acid tolunic acid
  • the cockroach extract is a hull extract in which the total content of ruthenium acid A and ruthenium acid B is not less than 40% by weight based on the total weight of the husk extract.
  • the preparation is provided in the form of a cosmetic, a skin care product or a food, and the skin is maintained by enhancing the expression of collagen and/or hyaluronic acid.
  • the cosmetics, skin care products and foods are used for at least one of the following: moisturizing the skin, firming the skin, repairing the skin, moisturizing, diminishing wrinkles, delaying the aging of the skin, and preventing skin aging.
  • the preparation is provided in the form of a medicament and promotes wound healing by enhancing the expression of collagen and/or hyaluronic acid.
  • Another object of the present invention is to provide a use of an active ingredient for the preparation of a preparation for the maintenance of skin and/or for promoting wound healing, wherein the active ingredient is selected from at least one of the following: dehydroabietic acid, Capric acid, dehydromomoic acid, turonic acid, porcine acid C, 3-epoxy-dehydroglutenic acid, dehydroabietic acid, tradulinic acid, dehydroporosic acid, porphyric acid, Fuxin acid A and ruthenium acid B.
  • the active ingredient, ruthenium acid A and/or ruthenium acid B is provided in the form of an extract, especially in the form of a fungal extract or a plant extract.
  • the above preparation is provided in the form of a cosmetic, a skin care product, a food or a medicament, wherein the medicament promotes wound healing by enhancing the performance of collagen and/or hyaluronic acid, the cosmetic, skin care and food enhancing collagen Protein and/or hyaluronic acid to maintain skin for at least one of the following: moisturizes skin, tightens skin, repairs skin, moisturizes, diminishes wrinkles, delays skin aging, and prevents skin aging.
  • compositions for maintaining skin and/or promoting wound healing which composition is a cosmetic, skin care product, food or medicament, and comprises an effective amount of the above active ingredient or quinone extract.
  • a refers to the amount of compound which can effectively enhance the expression of collagen and/or hyaluronic acid of the individual when administered to an individual;
  • the so-called "individual” means a mammal, and the mammal can be a human or a non-human animal;
  • /kg body weight refers to the dose required per kilogram of body weight.
  • Collagen is known to be an important glycoprotein in the connective tissue and extracellular matrix of the human body, accounting for more than 20% of the total protein of the human body. Collagen is entangled into a triple helix with three peptide chains as a basic structure. After being synthesized in skin fibroblasts, this triple helix is secreted outside the cell to form tight mature collagen fibers, which gives collagen a skin elasticity. Improve skin's water retention and promote wound healing.
  • Hyaluronic acid is abundant in the dermis layer and belongs to mucopolysaccharides. It has a high moisturizing function, which can keep the skin elastic and moist, reduce wrinkles and prevent skin aging.
  • the medicinal material refers to the dried sclerotium of the fungus fungus (Poria cocos (Schw.) Wolf).
  • the fungus is often parasitic on the roots of pine trees.
  • the outer skin is light brown or dark brown (skin), and the interior is pink or white (clam meat).
  • Traditional Chinese medicine records contain sedative, diuretic, nutrient supplementation, immunity enhancement and delay aging. Modern medical research has also confirmed that cockroaches have anti-inflammatory, anti-tumor, immune system regulation, blood sugar lowering, and nutrient absorption.
  • sputum extract can effectively promote the expression of collagen and / or hyaluronic acid (ie, enhance the content of endogenous collagen and / or hyaluronic acid), so it can be used to maintain the skin and promote wound healing.
  • the present invention provides an use of a sputum extract for the maintenance of skin and/or for promoting wound healing, including the use of a sputum extract to prepare a preparation for skin care and/or promoting wound healing, for an individual in need thereof A method of administering a sputum extract to care for the skin and/or promoting wound healing, and providing a composition comprising the sputum extract.
  • the ruthenium extract used may be a crude extract or extract provided by an operation comprising the steps of: (a) extracting the ruthenium raw material with a first polar solvent to obtain a crude extract; b) drying the crude extract to obtain a crude extract powder; (c) extracting the crude extract powder with a second polar solvent to obtain a cerium extract.
  • the first polar solvent is the same as or different from the second polar solvent, and may be selected from the group consisting of water, ethanol, and a combination thereof.
  • an aqueous ethanol solution having the same or different ethanol concentration is used as the first polar solvent and the second polar solvent.
  • the niobium material may be a suede portion and/or a clam portion, and the ratio of the amount of the first polar solvent to the niobium material may be adjusted as needed.
  • the amount of the first polar solvent to be used is not particularly limited as long as the raw materials can be uniformly dispersed.
  • the volume ratio of the first polar solvent to the ruthenium raw material may be employed in step (a) from about 8:1 to about 16:1.
  • the extracting step (a) is carried out by using the suede as a raw material, and the aqueous ethanol solution is the first polar solvent, and the volume ratio of the aqueous ethanol: raw material is about 8:1.
  • the suede portion and the meat portion are used as raw materials, and the aqueous ethanol solution is the first polar solvent, and the volume ratio of the aqueous ethanol solution: raw material is about 8:1. Extraction of step (a).
  • a suitable extraction time can be selected depending on the first polar solvent employed. For example, using an aqueous solution of ethanol as the first polar solvent and a volume ratio of aqueous ethanol: hydrazine of 8:1, the extraction is usually carried out for at least 1 hour, preferably at least 2 hours, more preferably at least 3 hours. Further, other operations such as boiling, cooling, filtration, concentration under reduced pressure, resin column chromatography, and the like may be additionally performed to carry out the step (a). Alternatively, it may be necessary to pre-soak the crucible in the first polar solvent for a period of time prior to performing step (a). For example, using an aqueous solution of ethanol as the first polar solvent, pre-soaking may be performed for, for example, about 12 hours.
  • the ratio of the amount of the second polar solvent to the crude extract powder obtained in the step (b) may be adjusted as needed.
  • the amount of the second polar solvent to be used is not particularly limited as long as the coarse extract powder can be uniformly dispersed.
  • the volume ratio of the second polar solvent to the crude extract powder may be employed in step (c) from about 8:1 to about 16:1.
  • the extraction of the step (c) is carried out by using an aqueous solution of ethanol as the second polar solvent and a volume ratio of the aqueous ethanol solution: the crude extract powder of about 8:1.
  • the hydrazine extract used in accordance with the present invention may also be a dry product which may be provided by drying the extract obtained in step (c).
  • the hydrazine may be repeatedly extracted with the same or different first polar solvent before the step (b), and the extract obtained by the multiple extraction may be combined to provide
  • the crude extract of step (b) is carried out; the cycles of step (b), step (c), and other operations as desired may also be repeated.
  • hydroquinone acid, citric acid, dehydromomoic acid, turonic acid, porcine acid C, 3-epoxy-dehydromethane can be isolated and purified from the hydrazine extract.
  • the molting extract provided according to the present invention contains dehydromomoic acid, porcine cyanate C, 3-epoxy-dehydroglutenic acid, dehydroabietic acid, ceric acid, dehydroporosporium Acid, layer of porphyric acid, ruthenium acid A, and ruthenium acid B and other ingredients.
  • the inventors of the present invention further studied and found that dehydroabietic acid, citric acid, dehydromomoic acid, turonic acid, porcine acid C, 3-epoxy-dehydroglutenic acid, dehydroabietic acid, tyrosinic acid, Dehydroporosporin, streptococcus, ruthenium A and ruthenium B can effectively promote the expression of collagen and/or hyaluronic acid (ie, enhance endogenous collagen and/or hyaluronic acid). It can be used to maintain skin and promote wound healing.
  • the present invention is also directed to an use of an active ingredient for the maintenance of skin and/or for promoting wound healing, wherein the active ingredient is selected from at least one of the following: dehydroabietic acid, citric acid, dehydrohydro acid, Tolunic acid, porcine acid C, 3-epoxy-dehydroglutenic acid, dehydroabietic acid, tyloscinic acid, dehydroporosporin acid, porphyrin acid, ruthenium acid A and ruthenium acid B .
  • the active ingredient is selected from at least one of the following: dehydroabietic acid, citric acid, dehydrohydro acid, Tolunic acid, porcine acid C, 3-epoxy-dehydroglutenic acid, dehydroabietic acid, tyloscinic acid, dehydroporosporin acid, porphyrin acid, ruthenium acid A and ruthenium acid B .
  • the foregoing applications include the use of the active ingredient to prepare a preparation for the maintenance of the skin and/or to promote wound healing, the administration of the active ingredient to an individual in need thereof to maintain the skin and/or to promote wound healing, and the provision of a method A composition comprising the active ingredient.
  • the active ingredient employed in accordance with the invention is provided in the form of an extract; more preferably, the active ingredient is provided in the form of a fungal extract or a plant extract.
  • the formulation or composition can be provided in the form of a cosmetic, skin care, food, or pharmaceutical.
  • the cosmetic or skin care product may be in any convenient form, and is not particularly limited, and is in a correspondingly suitable dosage form depending on the intended use.
  • the cosmetic or skin care product may be an emulsion, a cream, a gel (such as a hydrogel), a paste (such as a dispersion cream, an ointment), a spray, or a spray for direct external use. Or a solution (eg, lotion, suspension) or the like.
  • the preparation of the invention or The composition is prepared in the form of a food product that can be swallowed or consumed, such as a health food, a beauty drink, and the like.
  • the formulations or compositions of the invention may also be provided in the form of a subcutaneous injection.
  • the preparation or composition according to the present invention when the preparation or composition according to the present invention is provided in the form of a medicament, the medicament may be in any convenient form, and is not particularly limited, and is in a correspondingly suitable dosage form depending on the intended use.
  • the agent can be administered orally or non-orally (eg, transdermally, subcutaneously, or intravenously) to an individual in need thereof to maintain the skin and/or promote wounds. Heal.
  • a suitable carrier may be selected to provide the agent, wherein the carrier includes an excipient, a diluent, an adjuvant, a stabilizer, an absorption delaying agent, a disintegrating agent, a solubilizing agent, Emulsifiers, antioxidants, binders, binders, tackifiers, dispersants, suspending agents, lubricants, and/or moisture absorbents, and the like.
  • the pharmaceutical agent may contain any active ingredient which does not adversely affect the active ingredient (i.e., strontium extract, dehydroabietic acid, citric acid, dehydrochloric acid, toluic acid, At least one of porcine citrate C, 3-epoxy-dehydroglutenic acid, dehydroabioric acid, ceric acid, dehydroporosic acid, porphyric acid, ruthenium acid A, and ruthenium acid B a pharmaceutically acceptable carrier of the desired benefit, which carrier can, for example, be selected from the group consisting of water, saline, dextrose, glycerol, ethanol or the like, cellulose, starch, sugar bentonite (sugar bentonite), and combinations of the foregoing.
  • active ingredient i.e., strontium extract, dehydroabietic acid, citric acid, dehydrochloric acid, toluic acid, At least one of porcine citrate C, 3-epoxy-dehydroglutenic acid, de
  • the agent may be provided in a dosage form suitable for oral administration by any convenient method, for example, a tablet (for example, a sugar-coated tablet), a pill, a capsule, a granule, a powder, a flow extract, a solution, a syrup, a suspension. Agents, tinctures, etc.
  • an injection or a drip dosage form suitable for subcutaneous or intravenous administration one or more e.g. isotonic solutions, a salt buffer (such as a phosphate buffer or a citrate buffer), a solubilizing agent, or the like may be contained in the medicament.
  • the medicament is provided in a dosage form such as an injection, a dry powder injection, a suspension injection, or a dry powder suspension injection.
  • the medicament is prepared as a pre-injection solid
  • the pre-injection solid is provided in a dosage form, or emulsifiable, which is soluble in the other solution or suspension, and prior to injection, prior to administration to an individual in need thereof.
  • the solid is dissolved in other solutions or suspensions or emulsified to provide the desired injection.
  • a suitable amount of an additive may be additionally included in the preparation or composition provided according to the present invention, for example, a flavoring agent, a toner, or a flavoring agent which enhances the mouthfeel and visual sensation of the preparation or composition when administered.
  • a coloring agent or the like, and a buffering agent, a preservative, a preservative, an antibacterial agent, an antifungal agent and the like which can improve the stability and storage property of the preparation or composition.
  • the preparation or composition may additionally contain one or more other active ingredients (for example, collagen, hyaluronic acid, elastin, mandelic acid, arbutin, etc.), or may contain the one or more other active ingredients.
  • the care product or drug is used in combination to further enhance the efficacy of the formulation or composition or to increase the flexibility and formulation of the formulation or composition formulation. In principle, as long as the other ingredients or additives added do not adversely affect the desired efficacy of the formulations or compositions of the invention.
  • formulations or compositions provided for use in accordance with the present invention may be administered at different frequencies, such as once a day, multiple times a day, or several times a day, depending on the individual's needs, age, weight, condition, and purpose of administration. Different.
  • the invention also provides a method for the maintenance of skin and/or to promote wound healing comprising administering to an individual in need thereof an effective amount of the active ingredient (ie, sputum extract, dehydroabietic acid, citric acid, Dehydromomoic acid, toluic acid, porcine acid C, 3-epoxy-dehydroglutenic acid, dehydroabietic acid, oxytetracycline, dehydroporosic acid, porphyric acid, ruthenium acid At least one of A and ruthenium acid B).
  • the active ingredient ie, sputum extract, dehydroabietic acid, citric acid, Dehydromomoic acid, toluic acid, porcine acid C, 3-epoxy-dehydroglutenic acid, dehydroabietic acid, oxytetracycline, dehydroporosic acid, porphyric acid, ruthenium acid At least one of A and ruthenium acid B).
  • A-1 Take the medicinal herbs (source origin is Yunnan), peel off the outer skin after washing (hereinafter referred to as “skin”), and the rest is the meat department (hereinafter referred to as “the meat department”).
  • A-2 Take the mink obtained from A-1, soak it in 75% ethanol solution at a volume ratio of 1:8 (medicine: ethanol solution) at room temperature for 12 hours, then boil and extract. (It lasted 3 hours). The aforementioned extraction steps were repeated for a total of three times. The resulting extract was combined three times and filtered to remove insolubles to obtain a crude extract. Next, the crude extract described above was concentrated under reduced pressure to remove the solvent, and then dried by a spray dryer to obtain a crude extract powder.
  • A-3 taking the crude extract powder obtained in A-2, mixing it with 95% ethanol in a volume ratio of 1:8 (crude extract powder: aqueous ethanol solution), and extracting (for 3 hours), and then, Separation was carried out using a column packed with silica gel as a stationary phase to obtain a suede extract.
  • the cockroach extract contained dehydroabietic acid, citric acid, dehydromomoic acid, turmeric acid, porcine acid C, 3-epoxy-dehydromethane acid, dehydroabietic acid, oleic acid. , dehydrogenated layer acid, layer of porphyric acid, ruthenium acid A and ruthenium B.
  • the cerium extract obtained in A-5 was uniformly dissolved in methanol at a volume ratio of 1:500 (extract: methanol). After filtering and removing insoluble matter, a preparative high-performance liquid chromatograph (mixed with methanol and water as mobile phase) for dehydroquinone at 243 nm and 210 nm respectively Citrate, citric acid, dehydromomoic acid, turonic acid, porcine acid C, 3-epoxy-dehydroglutenic acid, dehydroabietic acid, tylosic acid, dehydroporosporin, layer pore The phytic acid, ruthenium acid A and ruthenium acid B were separated and collected.
  • B-2 Dehydroabietic acid, citric acid, dehydrohydrogen acid, and toluic acid obtained by B-1 were detected by liquid chromatography/ultraviolet/mass spectrometry at 243 nm and 210 nm respectively.
  • Choline citrate C 3-epoxy-dehydroglutenic acid, dehydrochaetidic acid, ceric acid, dehydroporosporin acid, porphyrin acid, ruthenium acid A and ruthenium acid B, the results show that The purity of the ingredients is greater than 98%.
  • Example 1 Effect of lanthanum extract on the ability of cells to express collagen and hyaluronic acid
  • the normal human skin primary fibroblasts provided in [Preparation Example C] were taken, divided into four groups and cultured in the following medium for 48 hours:
  • Group I fibroblast growth medium
  • Group II a fibroblast growth medium containing 0.005 ⁇ g/ml [Preparation Example A-3] provided by the molting extract;
  • Group III a fibroblast growth medium containing 0.05 ⁇ g/ml of the ecdysis extract provided in [Preparation Example A-3];
  • Group IV Fibroblast growth medium containing 0.5 ⁇ g/ml of the ecdysis extract provided in [Preparation Example A-3].
  • each group of cell culture liquid was collected and concentrated, and then analyzed by Western blotting method and enzyme immunoassay (ELISA) to determine the intracellular collagen content of each group and the cell culture medium (ie, extracellular The content of collagen and the content of hyaluronic acid in the cell culture solution.
  • ELISA enzyme immunoassay
  • the relative content (in percentage) of collagen and hyaluronic acid of the other groups was calculated by the control group (i.e., the cells cultured in the group I culture medium), and the results are shown in Table 2.
  • Group II Group III Group IV Intracellular collagen (%) 152 182 200 Collagen in cell culture medium (%) 116 156 135 Hyaluronic acid (%) in cell culture medium 131 121 106
  • the intracellular collagen content of the cells treated with the sputum extract of the present invention i.e., the cells cultured in the culture medium of Group II, III, or IV
  • the control group in the cell culture medium. That is, the extracellular collagen content and the hyaluronic acid content in the cell culture solution are significantly improved.
  • the foregoing results show that the cockroach extract of the present invention can effectively enhance the ability of the fibroblast to express collagen and/or hyaluronic acid (ie, increase the content of endogenous collagen and/or hyaluronic acid), so it can be used for skin care. And / or promote wound healing.
  • Example 2 Effect of active ingredients of sputum extract on the ability of cells to express collagen and hyaluronic acid
  • Group 1 Fibroblast growth medium
  • Groups 2-1 to 2-3 a fibroblast growth medium containing the citric acid (PA) provided in [Preparation Example B] at concentrations of 0.01, 0.1, and 1 ⁇ mol, respectively;
  • PA citric acid
  • Groups 3-1 to 3-3 a fibroblast growth medium containing dehydroabietic acid (DPA) provided in [Preparation Example B] at concentrations of 0.01, 0.1, and 1 ⁇ mol, respectively;
  • DPA dehydroabietic acid
  • Groups 4-1 to 4-3 a fibroblast growth medium containing the toluic acid (TA) provided in [Preparation Example B] at concentrations of 0.01, 0.1, and 1 ⁇ mol, respectively;
  • Groups 5-1 to 5-3 Fibroblast growth medium containing dehydrochloric acid (DTA) supplied in [Preparation Example B] at concentrations of 0.01, 0.1, and 1 micromolar, respectively ;
  • DTA dehydrochloric acid
  • Groups 6-1 to 6-3 a fibroblast growth medium containing the porcine cyanic acid C (PAC) provided in [Preparation Example B] at concentrations of 0.01, 0.1, and 1 micromolar, respectively;
  • PAC porcine cyanic acid C
  • Groups 7-1 to 7-3 a fibroblast growth medium containing 3-table-dehydroglutenic acid (EDTA) provided in [Preparation Example B] at concentrations of 0.01, 0.1, and 1 micromolar concentration;
  • EDTA 3-table-dehydroglutenic acid
  • Groups 8-1 to 8-3 Fibroblast growth medium containing dehydroabietic acid (DTTA) provided in [Preparation Example B] at concentrations of 0.01, 0.1, and 1 micromolar, respectively ;
  • DTTA dehydroabietic acid
  • Groups 9-1 to 9-3 a fibroblast growth medium containing the thrombic acid (TTA) provided in [Preparation Example B] at concentrations of 0.01, 0.1, and 1 ⁇ mol, respectively;
  • TTA thrombic acid
  • Groups 10-1 to 10-3 a fibroblast growth medium containing the ruthenium acid A (PAA) provided in [Preparation Example B] at concentrations of 0.01, 0.1, and 1 micromolar, respectively;
  • PAA ruthenium acid A
  • Groups 11-1 to 11-3 a fibroblast growth medium containing dehydroporosporin (DEA) provided in [Preparation Example B] at concentrations of 0.01, 0.1, and 1 micromolar, respectively.
  • DEA dehydroporosporin
  • Groups 12-1 to 12-3 a fibroblast growth medium containing the ruthenium acid B (PAB) provided in [Preparation Example B] at concentrations of 0.01, 0.1, and 1 ⁇ mol, respectively;
  • PAB ruthenium acid B
  • Groups 13-1 to 13-3 A fibroblast growth medium containing the layer of porcine acid (EA) supplied in [Preparation Example B] at concentrations of 0.01, 0.1, and 1 micromolar, respectively.
  • EA porcine acid
  • each group of cell culture liquids was collected and concentrated, and then analyzed by Western blotting method and enzyme immunoassay (ELISA) to determine the intracellular collagen content of each group.
  • ELISA enzyme immunoassay
  • Example (2-1) the concentrated group of cell culture solutions provided in Example (2-1) were subjected to Western blotting and enzyme immunoassay (ELISA). For analysis, the collagen content in each group of culture fluid (ie, extracellular) was determined. Finally, the relative content (in percentage) of collagen of the other groups was calculated by the control group (i.e., the cells cultured in the first group culture medium) was 100%, and the results are shown in Table 4.
  • ELISA enzyme immunoassay
  • Example (2-1) the concentrated group of cell culture solutions provided in Example (2-1) were subjected to Western blotting and enzyme immunoassay (ELISA). An analysis was performed to determine the hyaluronic acid content in each group of culture fluids (ie, extracellular). Finally, the relative content of hyaluronic acid (expressed as a percentage) of the other groups was calculated as the result of the control group (i.e., the cells cultured in the first group culture medium), and the results are shown in Table 5.
  • ELISA enzyme immunoassay
  • the extract of the present invention and/or its active ingredient can indeed enhance the ability of cells to express collagen and hyaluronic acid (ie, the ability to enhance the content of endogenous collagen and hyaluronic acid) It can be used to maintain the skin (including moisturizing the skin, firming the skin, repairing the skin, moisturizing, diminishing wrinkles, delaying skin aging, and preventing skin aging) and/or promoting wound healing.

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Abstract

一种使用茯苓萃取物、去氢茯苓酸(DPA)、茯苓酸(PA)、去氢土莫酸(DTA)、土莫酸(TA)、猪苓酸C(PAC)、3-表-去氢土莫酸(EDTA)、去氢栓菌酸(DTTA)、栓菌酸(TTA)、去氢层孔菌酸(DEA)、层孔菌酸(EA)、茯苓新酸A(PAA)及/或茯苓新酸B(PAB)于制备一制剂的用途，其中该制剂用于保养肌肤及/或促进伤口愈合。

Description

茯苓萃取物及其活性成分于保养肌肤及/或促进伤口愈合的用途 技术领域
本发明关于茯苓萃取物及/或其活性成分的应用，尤其是关于使用茯苓萃取物及/或其活性成分以保养肌肤及/或促进伤口愈合的应用。其中，该茯苓萃取物及/或其活性成分通过提升胶原蛋白及/或透明质酸的表现以保养肌肤及/或促进伤口愈合。
背景技术
当肌肤的水分含量降低时，会使得肌肤表面的角质细胞因为无法正常脱落而堆积，进而使肌肤失去光泽与弹性，显得干燥、粗糙，甚至出现皱纹等肌肤老化现象。因此，若可有效提升肌肤的水分含量，将可减少前述情况的发生。
研究证实，胶原蛋白(collagen)与透明质酸(hyaluronic acid)具有提升肌肤水分含量的功能，故市面上许多添加有胶原蛋白及/或透明质酸的保养品、健康食品，都宣称可提供滋润肌肤、紧实肌肤、修复肌肤、保湿、淡化皱纹、延缓肌肤老化、及预防肌肤老化的效果。此外，胶原蛋白因具有高生物相容性、低免疫性，且可以被身体分解吸收，故也有健康食品因添加有胶原蛋白而宣称可提供促进伤口愈合的效果。
然而，近年来在世界各地爆发口蹄疫、禽流感、狂牛症等人畜共通传染病，此使得动物性胶原蛋白及透明质酸的安全性受到质疑。此外，不论是以口服、涂抹或是注射的方式以使用外源性胶原蛋白或透明质酸，都有吸收率不佳、或是可能引起免疫反应的问题。此所以，业界对于外源性胶原蛋白与透明质酸的使用，一直以来都存在相当大的争议。因此，业界目前仍积极开 发可有效提升内源性胶原蛋白及/或透明质酸表现量的制剂或方法，希望通过内源性胶原蛋白及/或透明质酸的提升以达到保养肌肤及/或促进伤口愈合的效果，且避免因补充外源性胶原蛋白及/或透明质酸而引起的安全性及有效性问题。
本案发明人研究发现，茯苓萃取物及/或其活性成分可有效提升细胞内胶原蛋白、细胞外胶原蛋白、及/或细胞外透明质酸的含量，此即，茯苓萃取物及/或其活性成分可有效提升内源性胶原蛋白及/或透明质酸的含量，故可用于保养肌肤及/或促进伤口愈合。
发明内容
因此，本发明的一目的，在于提供一种使用茯苓萃取物于制备一制剂的用途，其中该制剂用于保养肌肤及/或促进伤口愈合。较佳地，该茯苓萃取物包含以下的至少一者：去氢茯苓酸(dehydropachymic acid，DPA)、茯苓酸(pachymic acid，PA)、去氢土莫酸(dehydrotumulosic acid，DTA)、土莫酸(tumulosic acid，TA)、猪苓酸C(polyporenic acid C，PAC)、3-表-去氢土莫酸(3-epi-dehydrotumulosic acid，EDTA)、去氢栓菌酸(dehydrotrametenolic acid，DTTA)、栓菌酸(trametenolic acid，TTA)、去氢层孔菌酸(dehydroeburicoic acid，DEA)、层孔菌酸(eburicoic acid，EA)、茯苓新酸A(poricoic acid A，PAA)及茯苓新酸B(poricoic acid B，PAB)。更佳地，该茯苓萃取物为茯苓皮部萃取物，其中以茯苓皮部萃取物的总重量计，茯苓新酸A及茯苓新酸B的总含量不小于40重量％。
其中，该制剂以一化妆品、保养品或食品的形式提供，且通过提升胶原蛋白及/或透明质酸的表现以保养肌肤。
其中，该化妆品、保养品及食品用于以下的至少一者：滋润肌肤、紧实肌肤、修复肌肤、保湿、淡化皱纹、延缓肌肤老化、及预防肌肤老化。
其中，该制剂以一药剂的形式提供，且通过提升胶原蛋白及/或透明质酸的表现以促进伤口愈合。
本发明的另一目的，在于提供一种使用一活性成分于制备一用于保养肌肤及/或促进伤口愈合的制剂的用途，其中该活性成分选自以下的至少一者：去氢茯苓酸、茯苓酸、去氢土莫酸、土莫酸、猪苓酸C、3-表-去氢土莫酸、去氢栓菌酸、栓菌酸、去氢层孔菌酸、层孔菌酸、茯苓新酸A及茯苓新酸B。较佳地，该活性成分茯苓新酸A及/或茯苓新酸B。更佳地，该活性成分以萃取物的形式提供，尤其是以真菌萃取物或植物萃取物的形式提供。
较佳地，上述制剂以一化妆品、保养品、食品或药剂的形式提供，其中，该药剂通过提升胶原蛋白及/或透明质酸的表现以促进伤口愈合，该化妆品、保养品及食品提升胶原蛋白及/或透明质酸的表现以保养肌肤，用于以下的至少一者：滋润肌肤、紧实肌肤、修复肌肤、保湿、淡化皱纹、延缓肌肤老化、及预防肌肤老化。
本发明的又一目的，在于提供一种保养肌肤及/或促进伤口愈合的方法，其包含对一有需要的个体投予一有效量的上述活性成分或茯苓萃取物。
本发明的再一目的，在于提供一种保养肌肤及/或促进伤口愈合的组合物，该组合物为一化妆品、保养品、食品或药剂，且包含一有效量的上述活性成分或茯苓萃取物。
本发明的详细技术内容及部分具体实施例，将描述于以下内容中，以供本领域技术人员据以明了本发明的特征。
具体实施方式
以下将描述根据本发明的部分具体实施例；但在不背离本发明精神下，本发明还可以多种不同形式的实施例来实践，不应将本发明保护范围解释为限于说明书所陈述的。此外，除非文中有另外说明，于本说明书中(尤其是在前述权利要求书中)所使用的“一”、“该”及类似用语应理解为包含单数及复数形式；所谓“有效量”，是指投予至个体时，可有效提升该个体的胶原蛋白及/或透明质酸的表现的化合物用量；所谓“个体”是指哺乳动物，哺乳动物可为人类或非人动物；单位“毫克/公斤体重”是指每公斤体重个体所须的剂量。
如上述说明，肌肤中的水分含量降低为造成肌肤老化的一个重要原因。已知胶原蛋白是人体的结缔组织与细胞外间质中一种重要的含醣蛋白质，占人体总蛋白的20％以上。胶原蛋白以三条胜肽链缠绕成三股螺旋作为基础结构，此三股螺旋结构在皮肤纤维母细胞内合成后，会被分泌到细胞外，形成紧密的成熟胶原纤维，此结构赋予胶原蛋白保持肌肤弹性、提高肌肤保水度及促进伤口愈合的功效。透明质酸则大量存在于真皮层中，属于粘多醣类，具有高度的保湿功能，可使肌肤保持弹性、湿润，并可减少皱纹、避免肌肤老化。
茯苓药材是指拟层孔菌科真菌(Poria cocos(Schw.)Wolf)的干燥菌核。茯苓真菌常寄生在松树根上，外皮呈淡棕色或黑褐色(茯苓皮部)，内部则呈粉红色或白色(茯苓肉部)。传统中医典籍记载茯苓具有镇静、利尿、营养素补充、免疫力增强及延缓老化等用途。现代医学研究也证实茯苓具有抗发炎、抗肿瘤、调节免疫系统、降血糖、促进营养吸收等功效。本案发明 人研究发现，茯苓萃取物可有效促进细胞表现胶原蛋白及/或透明质酸(即，提升内源性胶原蛋白及/或透明质酸的含量)，故可用于保养肌肤及促进伤口愈合。
因此，本发明提供一种使用茯苓萃取物于保养肌肤及/或促进伤口愈合的应用，包括使用茯苓萃取物制备一用于保养肌肤及/或促进伤口愈合的制剂的用途、对有需要的个体投予茯苓萃取物以保养肌肤及/或促进伤口愈合的方法、以及提供一包含茯苓萃取物的组合物。
根据本发明，所采用的茯苓萃取物可以是一通过包含如下步骤的操作所提供的粗萃物或萃取物：(a)以一第一极性溶剂萃取茯苓原料，获得一粗萃物；(b)干燥该粗萃物，获得一粗萃物粉末；(c)以一第二极性溶剂萃取该粗萃物粉末，获得茯苓萃取物。其中，该第一极性溶剂与该第二极性溶剂相同或不同，且可分别选自水、乙醇、及前述的组合。于本发明的部分实施例中，采用乙醇浓度相同或不同的乙醇水溶液作为第一极性溶剂与第二极性溶剂。
于步骤(a)中，该茯苓原料可以是茯苓皮部及/或茯苓肉部，且可视需要调整第一极性溶剂与茯苓原料的用量比率。一般而言，第一极性溶剂的用量并无特殊限制，只要可使原料均匀分散即可。举例言之，可于步骤(a)采用第一极性溶剂与茯苓原料的体积比为约8∶1至约16∶1的用量。于本发明一具体实施例中，以茯苓皮部作为原料，且乙醇水溶液为第一极性溶剂，并以乙醇水溶液∶原料的体积比为约8∶1的用量以进行步骤(a)的萃取。于本发明另一具体实施例中，以茯苓皮部及肉部作为原料，且乙醇水溶液为第一极性溶剂，并以乙醇水溶液∶原料的体积比为约8∶1的用量以进行步 骤(a)的萃取。
于步骤(a)中，可视所采用的第一极性溶剂来选用合宜的萃取时间。以采用乙醇水溶液作为第一极性溶剂且乙醇水溶液∶茯苓的体积比为8∶1为例，通常萃取历时至少1小时，较佳至少2小时，更佳至少3小时。此外，可视需要辅以例如煎煮、冷却、过滤、减压浓缩、树脂管柱层析等其它操作以进行步骤(a)。另外，可视需要于进行步骤(a)之前，先将茯苓预浸泡于第一极性溶剂中一段时间。以采用乙醇水溶液为第一极性溶剂为例，可先进行预浸泡历时例如约12小时。
于步骤(c)中，可视需要调整第二极性溶剂与由步骤(b)所获得的粗萃物粉末的用量比。一般而言，第二极性溶剂的用量并无特殊限制，只要可使粗萃物粉末均匀分散即可。举例言之，可于步骤(c)采用第二极性溶剂与茯苓粗萃物粉末的体积比为约8∶1至约16∶1的用量。于本发明一具体实施例中，以乙醇水溶液为第二极性溶剂，并以乙醇水溶液∶茯苓粗萃物粉末的体积比为约8∶1的用量以进行步骤(c)的萃取。
根据本发明所采用的茯苓萃取物，也可以是一干燥物，此干燥物可通过干燥步骤(c)所得的萃取液而提供。为尽可能达到最大的萃取效益，视需要地，可于进行步骤(b)之前，以相同或不同的第一极性溶剂对茯苓重复进行萃取，并合并该多次萃取所得的萃取液以提供进行步骤(b)的粗萃物；也可重复进行步骤(b)、步骤(c)、以及前述视需要的其它操作的循环。
如后附实施例所示，根据本发明，可由茯苓萃取物中纯化分离出去氢茯苓酸、茯苓酸、去氢土莫酸、土莫酸、猪苓酸C、3-表-去氢土莫酸、去氢栓菌酸、栓菌酸、去氢层孔菌酸、层孔菌酸、茯苓新酸A及茯苓新酸B等成分。 其中，根据本发明所提供的茯苓皮部萃取物含有去氢土莫酸、猪苓酸C、3-表-去氢土莫酸、去氢栓菌酸、栓菌酸、去氢层孔菌酸、层孔菌酸、茯苓新酸A、以及茯苓新酸B等成分。
本案发明人进一步研究发现，去氢茯苓酸、茯苓酸、去氢土莫酸、土莫酸、猪苓酸C、3-表-去氢土莫酸、去氢栓菌酸、栓菌酸、去氢层孔菌酸、层孔菌酸、茯苓新酸A及茯苓新酸B都可有效促进细胞表现胶原蛋白及/或透明质酸(即，提升内源性胶原蛋白及/或透明质酸的含量)，故可用于保养肌肤及促进伤口愈合。因此，本发明也关于一种使用一活性成分于保养肌肤及/或促进伤口愈合的应用，其中该活性成分选自以下的至少一者：去氢茯苓酸、茯苓酸、去氢土莫酸、土莫酸、猪苓酸C、3-表-去氢土莫酸、去氢栓菌酸、栓菌酸、去氢层孔菌酸、层孔菌酸、茯苓新酸A及茯苓新酸B。前述应用包括使用该活性成分以制备一用于保养肌肤及/或促进伤口愈合的制剂的用途、对有需要的个体投予该活性成分以保养肌肤及/或促进伤口愈合的方法、以及提供一包含该活性成分的组合物。较佳地，根据本发明所采用的活性成分以萃取物的形式提供；更佳地，该活性成分以真菌萃取物或植物萃取物的形式提供。
于根据本发明的应用，该制剂或组合物可以一化妆品、保养品、食品、或药剂的形式提供。当以一化妆品或保养品的形式提供该制剂或组合物时，该化妆品或保养品可呈任何合宜的形式，并无特殊的限制，视所欲的用途而呈对应的合宜剂型。举例言之，但不以此为限，该化妆品或保养品可呈供直接外用的乳液、乳霜、凝胶(例如水凝胶)、膏状物(例如分散膏、软膏)、喷雾剂、或溶液(例如洗液、悬浮液)等形式。或者，可将本发明的制剂或 组合物制备成可供吞食或饮用的食品形式，例如健康食品、美容饮品等。此外，也可将本发明制剂或组合物以皮下注射的针剂形式提供。
类似地，当以一药剂的形式提供该根据本发明的制剂或组合物时，该药剂可呈任何合宜的形式，并无特殊限制，视所欲的用途而呈对应的合宜剂型。举例言之，但不以此为限，该药剂可以口服或非经口服(例如：经皮、皮下、或静脉内)的投药方式施用至有需要的个体上，以保养肌肤及/或促进伤口愈合。其中，视使用形式及用途而定，可选用合宜的载剂以提供该药剂，其中，该载剂包括赋形剂、稀释剂、辅助剂、安定剂、吸收延迟剂、崩散剂、增溶剂、乳化剂、抗氧化剂、粘合剂、结合剂、增粘剂、分散剂、悬浮化剂、润滑剂、及/或吸湿剂等。
以适于口服投药的药剂形式为例，该药剂中可含有任何不会不利影响所含有活性成分(即，茯苓萃取物、去氢茯苓酸、茯苓酸、去氢土莫酸、土莫酸、猪苓酸C、3-表-去氢土莫酸、去氢栓菌酸、栓菌酸、去氢层孔菌酸、层孔菌酸、茯苓新酸A及茯苓新酸B的至少一者)的所欲效益的医药上可接受的载剂，该载剂可以例如选自以下群组：水、食盐水、葡萄糖(dextrose)、甘油、乙醇或其类似物、纤维素、淀粉、糖膨润土(sugar bentonite)、及前述的组合。可利用任何合宜的方法，以适于口服投药的剂型提供该药剂，例如：锭剂(例如糖衣锭)、丸剂、胶囊剂、颗粒剂、散剂、流浸膏剂、溶液剂、糖浆剂、悬液剂、酊剂等。
至于适于皮下或静脉内的针剂或点滴剂型，则可于该药剂中含有一或多种例如等张溶液、盐类缓冲液(如磷酸盐缓冲液或柠檬酸盐缓冲液)、增溶剂、乳化剂、5％糖溶液、以及其他载剂等成分，以静脉输注液、乳剂静脉输 注液、干粉注射剂、悬液注射剂、或干粉悬液注射剂等剂型提供该药剂。或者，将该药剂制备成一注射前固体，以可溶于其他溶液或悬浮液中的剂型、或可乳化的剂型提供该注射前固体，并于投予至有需要的个体之前，将该注射前固体溶于其他溶液或悬浮液中或将其乳化，以提供所欲的注射剂。
视需要地，可于根据本发明所提供的制剂或组合物中另外含有合宜用量的添加剂，例如可提高该制剂或组合物于服用时的口适感及视觉感受的调味剂、调色剂、着色剂等，以及可改善该制剂或组合物的稳定性及储存性的缓冲剂、保存剂、防腐剂、抗菌剂、抗真菌剂等。此外，该制剂或组合物可视需要另外含有一或多种其他活性成分(例如，胶原蛋白、透明质酸、弹性蛋白、杏仁酸、熊果素等)，或者与含有该一或多种其他活性成分的保养品或药物并用，以进一步加强该制剂或组合物的功效或增加该制剂或组合物配方的运用灵活性与调配度。原则上，只要所添加的其它成分或添加剂不会对本发明制剂或组合物的所欲功效没有不利的影响即可。
于根据本发明的应用所提供的制剂或组合物可以一日一次、一日多次、或数日一次等不同频率施用，视投予个体的需求、年龄、体重、健康况状、及施用目的而异。
本发明也提供一种用于保养肌肤及/或促进伤口愈合的方法，其包含对一有需要的个体投予一有效量的活性成分(即，茯苓萃取物、去氢茯苓酸、茯苓酸、去氢土莫酸、土莫酸、猪苓酸C、3-表-去氢土莫酸、去氢栓菌酸、栓菌酸、去氢层孔菌酸、层孔菌酸、茯苓新酸A及茯苓新酸B的至少一者)。于根据本发明的保养肌肤及/或促进伤口愈合的方法中，有关该活性成分的实施例、投予途径、投予形式、适用剂量、以及相关应用，均如上述的 说明。
现以下列实施例进一步例示说明本发明。其中这些实施例仅提供作为说明，而非用以限制本发明的保护范围。本发明的保护范围如权利要求书所示。
实施例
[制备实施例]
A.茯苓萃取物的制备
A-1.取茯苓药材(来源产地为云南)，清洗后剥取其外皮(下称“茯苓皮部”)，其余即为肉部(下称“茯苓肉部”)。
A-2.取A-1所获得的茯苓皮部，于室温下，以1∶8(茯苓药材∶乙醇水溶液)的体积比浸泡于75％乙醇水溶液中，历时12小时，然后煮沸并进行萃取(历时3小时)。重复前述萃取步骤，共三次。合并三次萃取所得的萃取液并过滤以去除不溶物，以获得一粗萃取物。接着，对前述的粗萃取物进行减压浓缩以去除溶剂，再以喷雾干燥机进行干燥，以获得一粗萃物粉末。
A-3.取A-2所获得的粗萃物粉末，以1∶8(粗萃物粉末∶乙醇水溶液)的体积比与95％乙醇混合，并进行萃取(历时3小时)，接着，再利用以硅胶为固定相的管柱进行分离，获得一茯苓皮部萃取物。
A-4.以液相层析/紫外光/质谱仪，分别于243纳米及210纳米波长下检测A-3所获得的茯苓皮部萃取物的成分，并以高效液相色谱法定量该萃取物中各成分的含量，分析结果示于表1。
表1
Figure PCTCN2017100526-appb-000002
A-5.合并A-1所获得的茯苓皮部及茯苓肉部，重复上述A-2及A-3的萃取步骤，以获得一茯苓萃取物。接着，以A-4的分析方法检测所获得的茯苓萃取物的成分。结果显示，该茯苓萃取物含有去氢茯苓酸、茯苓酸、去氢土莫酸、土莫酸、猪苓酸C、3-表-去氢土莫酸、去氢栓菌酸、栓菌酸、去氢层孔菌酸、层孔菌酸、茯苓新酸A及茯苓新酸B等成分。
B.茯苓萃取物的活性成分的制备
B-1.以1∶500(萃取物∶甲醇)的体积比，将A-5所获得的茯苓萃取物均匀溶解于甲醇中。过滤去除不溶物后，以制备级高效能液相层析仪(以甲醇与水混合作为移动相)，分别于243纳米及210纳米波长下，针对去氢茯 苓酸、茯苓酸、去氢土莫酸、土莫酸、猪苓酸C、3-表-去氢土莫酸、去氢栓菌酸、栓菌酸、去氢层孔菌酸、层孔菌酸、茯苓新酸A及茯苓新酸B进行分离及收集，收集后再以减压浓缩机去除甲醇，分别得到去氢茯苓酸、茯苓酸、去氢土莫酸、土莫酸、猪苓酸C、3-表-去氢土莫酸、去氢栓菌酸、栓菌酸、去氢层孔菌酸、层孔菌酸、茯苓新酸A及茯苓新酸B。
B-2.以液相层析/紫外光/质谱仪，分别于243纳米及210纳米波长下检测B-1所获得的去氢茯苓酸、茯苓酸、去氢土莫酸、土莫酸、猪苓酸C、3-表-去氢土莫酸、去氢栓菌酸、栓菌酸、去氢层孔菌酸、层孔菌酸、茯苓新酸A及茯苓新酸B，结果显示这些成分的纯度都大于98％。
C.细胞培养
将正常人类皮肤初代纤维母细胞(normal primary human skin fibroblastic cell，购自ATCC)培养在纤维母细胞生长培养基(Fibroblast Growth Medium)培养基中，以供后续实验使用。
实施例1：茯苓萃取物对细胞表现胶原蛋白及透明质酸的能力的影响
取[制备实施例C]所提供的正常人类皮肤初代纤维母细胞，将其分成四组并分别以如下培养基进行培养，历时48小时：
1.第I组：纤维母细胞生长培养基；
2.第II组：含有0.005微克/毫升[制备实施例A-3]所提供的茯苓皮部萃取物的纤维母细胞生长培养基；
3.第III组：含有0.05微克/毫升[制备实施例A-3]所提供的茯苓皮部萃取物的纤维母细胞生长培养基；
4.第IV组：含有0.5微克/毫升[制备实施例A-3]所提供的茯苓皮部萃取物的纤维母细胞生长培养基。
其后，收集各组细胞培养液并加以浓缩，再以西方墨点法及酵素免疫分析法(ELISA)进行分析，测定各组的细胞内胶原蛋白的含量、细胞培养液中(即，细胞外)胶原蛋白的含量、以及细胞培养液中透明质酸的含量。最后，以控制组(即，经第I组培养液培养的细胞)的结果为100％，计算其他各组的胶原蛋白及透明质酸的相对含量(以百分比表示)，结果示于表2。
表2
  第II组 第III组 第IV组
细胞内胶原蛋白(％) 152 182 200
细胞培养液中胶原蛋白(％) 116 156 135
细胞培养液中透明质酸(％) 131 121 106
由表2可知，相较于控制组，经本发明茯苓萃取物处理的细胞(即，以第II、III、或IV组培养液培养的细胞)的细胞内胶原蛋白含量、细胞培养液中(即，细胞外)胶原蛋白含量、以及细胞培养液中的透明质酸含量都显著提升。前述结果显示，本发明茯苓萃取物可有效提升纤维母细胞表现胶原蛋白及/或透明质酸的能力(即，提升内源性胶原蛋白及/或透明质酸的含量)，故可用于保养皮肤及/或促进伤口愈合。
实施例2：茯苓萃取物的活性成分对细胞表现胶原蛋白及透明质酸的能力的影响
(2-1)细胞内的胶原蛋白表现
取[制备实施例C]所提供的正常人类皮肤初代纤维母细胞，将其分成三 十七组，分别以如下培养基进行培养，历时48小时：
1.第1组：纤维母细胞生长培养基；
2.第2-1至2-3组：含有[制备实施例B]所提供的茯苓酸(PA)的纤维母细胞生长培养基，浓度分别为0.01、0.1、及1微摩尔浓度；
3.第3-1至3-3组：含有[制备实施例B]所提供的去氢茯苓酸(DPA)的纤维母细胞生长培养基，浓度分别为0.01、0.1、及1微摩尔浓度；
4.第4-1至4-3组：含有[制备实施例B]所提供的土莫酸(TA)的纤维母细胞生长培养基，浓度分别为0.01、0.1、及1微摩尔浓度；
5.第5-1至5-3组：含有[制备实施例B]所提供的去氢土莫酸(DTA)的纤维母细胞生长培养基，浓度分别为0.01、0.1、及1微摩尔浓度；
6.第6-1至6-3组：含有[制备实施例B]所提供的猪苓酸C(PAC)的纤维母细胞生长培养基，浓度分别为0.01、0.1、及1微摩尔浓度；
7.第7-1至7-3组：含有[制备实施例B]所提供的3-表-去氢土莫酸(EDTA)的纤维母细胞生长培养基，浓度分别为0.01、0.1、及1微摩尔浓度；
8.第8-1至8-3组：含有[制备实施例B]所提供的去氢栓菌酸(DTTA)的纤维母细胞生长培养基，浓度分别为0.01、0.1、及1微摩尔浓度；
9.第9-1至9-3组：含有[制备实施例B]所提供的栓菌酸(TTA)的纤维母细胞生长培养基，浓度分别为0.01、0.1、及1微摩尔浓度；
10.第10-1至10-3组：含有[制备实施例B]所提供的茯苓新酸A(PAA)的纤维母细胞生长培养基，浓度分别为0.01、0.1、及1微摩尔浓度；
11.第11-1至11-3组：含有[制备实施例B]所提供的去氢层孔菌酸(DEA)的纤维母细胞生长培养基，浓度分别为0.01、0.1、及1微摩尔浓度；
12.第12-1至12-3组：含有[制备实施例B]所提供的茯苓新酸B(PAB)的纤维母细胞生长培养基，浓度分别为0.01、0.1、及1微摩尔浓度；
13.第13-1至13-3组：含有[制备实施例B]所提供的层孔菌酸(EA)的纤维母细胞生长培养基，浓度分别为0.01、0.1、及1微摩尔浓度。
其后，收集各组细胞培养液并加以浓缩，再以西方墨点法及酵素免疫分析法(ELISA)进行分析，测定各组的细胞内胶原蛋白的含量。最后，以控制组(即，经第1组培养液培养的细胞)的结果为100％，计算其他各组的胶原蛋白的相对含量(以百分比表示)，结果示于表3。
表3
Figure PCTCN2017100526-appb-000003
Figure PCTCN2017100526-appb-000004
由表3可知，相较于控制组，经茯苓酸、去氢茯苓酸、土莫酸、去氢土莫酸、猪苓酸C、去氢栓菌酸、或茯苓新酸A处理的细胞，其细胞内的胶原蛋白的含量都显著提升。前述结果显示，茯苓酸、去氢茯苓酸、土莫酸、去氢土莫酸、猪苓酸C、去氢栓菌酸、及茯苓新酸A可有效提升纤维母细胞表现胶原蛋白的能力(即，提升内源性胶原蛋白及/或透明质酸的含量)，故可用于保养皮肤及/或促进伤口愈合。
(2-2)细胞外的胶原蛋白表现
为了解本发明活性成分对细胞外的胶原蛋白表现的影响，以西方墨点法及酵素免疫分析法(ELISA)，对实施例(2-1)所提供的经浓缩的各组细胞培养液进行分析，测定各组培养液中(即，细胞外)的胶原蛋白含量。最后，以控制组(即，经第1组培养液培养的细胞)的结果为100％，计算其他各组的胶原蛋白的相对含量(以百分比表示)，结果示于表4。
表4
Figure PCTCN2017100526-appb-000005
Figure PCTCN2017100526-appb-000006
由表4可知，相较于控制组，经去氢茯苓酸、茯苓酸、去氢土莫酸、土莫酸、猪苓酸C、3-表-去氢土莫酸、去氢栓菌酸、栓菌酸、去氢层孔菌酸、层孔菌酸、茯苓新酸A、及茯苓新酸B处理的细胞，其细胞外的胶原蛋白的含量都显著提升。前述结果显示，本发明活性成分可有效提升纤维母细胞表现及分泌胶原蛋白的能力，故可用于保养皮肤及/或促进伤口愈合。
(2-3)细胞外的透明质酸表现
为了解本发明活性成分对细胞外的透明质酸表现的影响，以西方墨点法及酵素免疫分析法(ELISA)，对实施例(2-1)所提供的经浓缩的各组细胞培养液进行分析，测定各组培养液中(即，细胞外)的透明质酸含量。最后，以控制组(即，经第1组培养液培养的细胞)的结果为100％，计算其他各组的透明质酸的相对含量(以百分比表示)，结果示于表5。
表5
Figure PCTCN2017100526-appb-000007
由表5可知，相较于控制组，经去氢茯苓酸、茯苓酸、去氢土莫酸、土莫酸、猪苓酸C、3-表-去氢土莫酸、栓菌酸、去氢层孔菌酸、层孔菌酸、茯苓新酸A或茯苓新酸B处理的细胞，其细胞外的透明质酸的含量都显著提升，其中，经茯苓酸、去氢茯苓酸、土莫酸、去氢土莫酸、及3-表-去氢土莫酸处理的细胞的提升效果特别显著。前述结果显示，本发明活性成分可有效提升纤维母细胞表现透明质酸的能力，故可用于保养皮肤。
如上述实施例所示，本发明茯苓萃取物及/或其活性成分确实可提升细胞表现胶原蛋白及透明质酸的能力(即，具有提升内源性胶原蛋白及透明质酸的含量的能力)，可用于保养皮肤(包括滋润肌肤、紧实肌肤、修复肌肤、保湿、淡化皱纹、延缓肌肤老化、及预防肌肤老化)及/或促进伤口愈合。

Claims (14)

  1. 一种使用茯苓萃取物于制备一制剂的用途，其特征在于：该制剂用于保养肌肤及/或促进伤口愈合。
  2. 如权利要求1所述的用途，其特征在于：该茯苓萃取物包含以下的至少一者：去氢茯苓酸(dehydropachymic acid，DPA)、茯苓酸(pachymic acid，PA)、去氢土莫酸(dehydrotumulosic acid，DTA)、土莫酸(tumulosic acid，TA)、猪苓酸C(polyporenic acid C，PAC)、3-表-去氢土莫酸(3-epi-dehydrotumulosic acid，EDTA)、去氢栓菌酸(dehydrotrametenolic acid，DTTA)、栓菌酸(trametenolic acid，TTA)、去氢层孔菌酸(dehydroeburicoic acid，DEA)、层孔菌酸(eburicoic acid，EA)、茯苓新酸A(poricoic acid A，PAA)及茯苓新酸B(poricoic acid B，PAB)。
  3. 如权利要求1所述的用途，其特征在于：该茯苓萃取物为茯苓皮部萃取物。
  4. 如权利要求3所述的用途，其特征在于：以茯苓皮部萃取物的总重量计，茯苓新酸A及茯苓新酸B的总含量不小于40重量％。
  5. 如权利要求1至4中任一项所述的用途，其特征在于：该制剂以一化妆品、保养品或食品的形式提供，且通过提升胶原蛋白及/或透明质酸的表现以保养肌肤。
  6. 如权利要求5所述的用途，其特征在于：该化妆品、保养品及食品用于以下的至少一者：滋润肌肤、紧实肌肤、修复肌肤、保湿、淡化皱纹、延缓肌肤老化、及预防肌肤老化。
  7. 如权利要求1至4中任一项所述的用途，其特征在于：该制剂以一药剂的形式提供，且通过提升胶原蛋白及/或透明质酸的表现以促进伤口愈 合。
  8. 一种使用一活性成分于制备一用于保养肌肤及/或促进伤口愈合的制剂的用途，其特征在于：该活性成分选自以下的至少一者：去氢茯苓酸、茯苓酸、去氢土莫酸、土莫酸、猪苓酸C、3-表-去氢土莫酸、去氢栓菌酸、栓菌酸、去氢层孔菌酸、层孔菌酸、茯苓新酸A及茯苓新酸B。
  9. 如权利要求8所述的用途，其特征在于：该活性成分为茯苓新酸A及/或茯苓新酸B。
  10. 如权利要求8所述的用途，其特征在于：该活性成分以萃取物的形式提供。
  11. 如权利要求10所述的用途，其特征在于：该活性成分以真菌萃取物或植物萃取物的形式提供。
  12. 如权利要求8至11中任一项所述的用途，其特征在于：该制剂以一化妆品、保养品或食品的形式提供，且通过提升胶原蛋白及/或透明质酸的表现以保养肌肤。
  13. 如权利要求12所述的用途，其特征在于：该化妆品、保养品及食品用于以下的至少一者：滋润肌肤、紧实肌肤、修复肌肤、保湿、淡化皱纹、延缓肌肤老化、及预防肌肤老化。
  14. 如权利要求8至11中任一项所述的用途，其特征在于：该制剂以一药剂的形式提供，且通过提升胶原蛋白及/或透明质酸的表现以促进伤口愈合。
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