WO2024251002A1 - 红景天苷的衍生物及制备方法和应用 - Google Patents

红景天苷的衍生物及制备方法和应用 Download PDF

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WO2024251002A1
WO2024251002A1 PCT/CN2024/095989 CN2024095989W WO2024251002A1 WO 2024251002 A1 WO2024251002 A1 WO 2024251002A1 CN 2024095989 W CN2024095989 W CN 2024095989W WO 2024251002 A1 WO2024251002 A1 WO 2024251002A1
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salidroside
hydroxy
preparation
reaction
solution
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French (fr)
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方诩
刘大明
殷文成
张爽
广濑芳彦
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Henrui Qingdao Biotech Co Ltd
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Henrui Qingdao Biotech Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
    • C07H15/18—Acyclic radicals, substituted by carbocyclic rings
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • 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
    • A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
    • 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
    • A61K8/60—Sugars; Derivatives thereof
    • A61K8/602—Glycosides, e.g. rutin
    • 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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
    • A61P25/16—Anti-Parkinson drugs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/06—Free radical scavengers or antioxidants
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/44—Preparation of O-glycosides, e.g. glucosides
    • 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
    • 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 a salidroside derivative, in particular to a hydroxy-alpha-salidroside compound.
  • Rhodiola rosea is a precious Chinese medicine.
  • the 2020 edition of the Chinese Pharmacopoeia includes the dried roots and rhizomes of Rhodiola rosea of the Crassulaceae family. It is produced in China and other places in China and grows in areas with an altitude of about 4050 to 5400 meters.
  • Drugs such as Rhodiola rosea tablets and Xinnaoxin capsules prepared from Rhodiola rosea have the effects of promoting blood circulation and removing blood stasis, dredge meridians and relieve pain.
  • the functions of health foods such as Rhodiola rosea capsules are to enhance immunity and relieve physical fatigue.
  • Salidroside is the effective active ingredient of the traditional Chinese medicine Rhodiola rosea.
  • Salidroside has physiological activities such as protecting cardiovascular and cerebrovascular vessels, anti-fatigue, anti-depression, anti-aging, anti-hypoxia, anti-radiation, anti-tumor, immune regulation, whitening and freckle removal.
  • Salidroside in its natural state plant-derived Salidroside
  • ⁇ -Salidroside glycosidic bond
  • ⁇ -type glycosidic bond
  • Chinese patent 202211359878.5 discloses an ⁇ -salidroside and its preparation method and application.
  • the ⁇ -salidroside has a chemical name: 2-(4-hydroxyphenyl)ethyl- ⁇ -D-glucoside, a molecular weight of 300.23, and a chemical formula as follows. It has excellent free radical scavenging activity and can be used as the main raw material for cosmetics, anti-fatigue health products and other products.
  • the applicant studied salidroside and its derivatives and obtained hydroxy- ⁇ -salidroside.
  • the present invention also provides a preparation method and application of hydroxy- ⁇ -salidroside.
  • the present invention provides a hydroxy- ⁇ -salidroside compound, chemical name: 2-(3,4-dihydroxyphenyl)ethyl- ⁇ -D-glucoside, chemical formula shown in Formula I, which has an ⁇ -glucoside bond on the alcoholic hydroxyl group of hydroxytyrosol.
  • the molecular weight of the hydroxy- ⁇ -salidroside of the present invention is 316.108 after LC-MS analysis.
  • the present invention provides a method for preparing the hydroxy- ⁇ -salidroside compound.
  • the preparation method comprises dissolving ⁇ -salidroside in a buffer solution, adding hydroxylase, and performing a hydroxylation reaction in a reaction system to prepare the salidroside.
  • the hydroxylase can be selected from commercially available hydroxylase preparations.
  • the commercially available hydroxylase enzyme preparations are derived from Merck & Co., Ltd., Shanghai McLean Biochemical Technology Co., Ltd. and Shanghai Yuanye Biotechnology Co., Ltd., including but not limited to monophenol monooxygenase (Monophenol monooxygenase), polyphenol oxidase (Polyphenol Oxidase), tyrosinase (Tyrosinase) and the like.
  • the reaction system contains ammonium salt.
  • the ammonium salt preferably includes but is not limited to ammonium chloride, ammonium acetate, or ammonium sulfate.
  • the present invention provides the use of the hydroxy- ⁇ -salidroside compound in the preparation of a product having a free radical scavenging function; or the use of the hydroxy- ⁇ -salidroside compound in cosmetics having a whitening function.
  • the free radical is a 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH) free radical.
  • DPPH 1,1-diphenyl-2-trinitrophenylhydrazine
  • the product is a cosmetic having the function of scavenging free radicals.
  • the product with free radical scavenging function is a functional food.
  • the present invention further provides a composition containing hydroxy- ⁇ -salidroside.
  • the composition has the function of scavenging free radicals or whitening skin.
  • the free radical is a DPPH free radical.
  • the cosmetic composition containing hydroxy- ⁇ -salidroside is selected from aqueous solutions, oils, emulsions, gel products, paste products, or others, and can be produced according to existing cosmetic technical specifications.
  • the functional food composition containing hydroxy- ⁇ -salidroside has a dosage form selected from capsules, tablets, pastes, aqueous solutions, or others, and can be produced according to existing functional food technical specifications.
  • the drugs for improving learning and memory disorders include but are not limited to drugs for treating Alzheimer's disease.
  • the drugs for improving learning and memory disorders include but are not limited to drugs for treating Parkinson's disease.
  • a method for whitening skin comprising applying an effective amount of hydroxy- ⁇ -salidroside or a composition comprising an effective amount of hydroxy- ⁇ -salidroside to the skin.
  • a method for improving learning and memory disorders comprising administering to a subject an effective amount of hydroxy- ⁇ -salidroside or a composition comprising an effective amount of hydroxy- ⁇ -salidroside.
  • the "subject" of the present invention refers to any organism that needs treatment, prevention or diagnosis, preferably a mammal, and more preferably a human.
  • Figure 1 TLC spectrum of the enzyme-catalyzed reaction solution of Example 1; in the figure, from left to right are lane 1 and lane 2; lane 1 is ⁇ -salidroside, and lane 2 is hydroxy- ⁇ -salidroside.
  • reagents or raw materials used in the present invention can be purchased through conventional channels. Unless otherwise specified, the reagents or raw materials used in the present invention are used in a conventional manner in the art or in accordance with the product instructions. In addition, any method and material similar to or equivalent to the described content can be applied to the method of the present invention. The preferred implementation methods and materials described in the text are for demonstration purposes only.
  • the hydroxy- ⁇ -salidroside of the present invention has a chemical name: 2-(3,4-dihydroxyphenyl)ethyl- ⁇ -D-glucoside, and its structural formula is shown in Formula I.
  • the preparation method of hydroxy- ⁇ -salidroside of the present invention uses ⁇ -salidroside as a substrate and realizes hydroxylation under the action of hydroxylase.
  • the hydroxylase may be a commercially available hydroxylase preparation from Merck & Co., Ltd., Shanghai MacLean Biochemical Technology Co., Ltd. and Shanghai Yuanye Biotechnology Co., Ltd., including catechol oxidase but not limited to monophenol monooxygenase, polyphenol oxidase, tyrosinase and the like.
  • the method of using the hydroxylase enzyme preparation includes adding it to the reaction system in the form of powder or liquid, or fixing the enzyme preparation on a resin to make an immobilized enzyme preparation and adding it to the reaction system.
  • the immobilized enzyme preparation can be reused.
  • the amount of hydroxylase and the reaction conditions have a great influence on the production efficiency, so it is crucial to select appropriate reaction conditions such as the amount of enzyme and reaction time.
  • the reaction can be carried out in the presence of a solvent, and the solvent used in the reaction can be any solvent as long as it does not affect the type and concentration range of the reaction, and specifically can include common reagents such as DMSO, 2-propanol, ethanol, etc. These solvents can be used alone or in combination of two or more.
  • the enzyme used in the present invention can be inactivated by heating or changing the pH value, thereby stopping the enzyme catalyzed reaction.
  • the mixture containing the hydroxy- ⁇ -salidroside compound, the hydrolyzate of the mixture, the enzyme-inactivated product of the mixture, the purified product of the mixture, and the dry powder product thereof can be used in foods, beverages and cosmetics containing the hydroxy- ⁇ -salidroside compound, and can also be used as an ingredient of food, special medical food, health products or medicines, and used in the production of food and beverages, cosmetics, special medical food, health products or medicines.
  • HPLC analysis conditions High performance liquid chromatograph: Hitachi HPLC 5440 chromatograph, chromatographic column: Kromasil 100-5C18 (250*4.6mm), detector: photodiode array (DAD 280nm), detection wavelength: UV 280nm, injection volume: 10 ⁇ L, flow rate: 1mL/min, column temperature: 30°C, mobile phase: acetonitrile: 0.1% formic acid aqueous solution (v/v) isocratic elution.
  • DAD 280nm photodiode array
  • LC-MS analysis conditions chromatographic column: Kromasil 100-5C18 (250*4.6mm), detector: photodiode array (DAD 280nm), detection wavelength: 280nm, injection volume: 10 ⁇ L, flow rate: 1mL/min, column temperature: 30°C, mobile phase with detection time of 5-15min; H-ESI mode, molecular weight scanning range 50 ⁇ 800.
  • the inventors of the present invention carried out acetylation treatment on the hydroxy- ⁇ -salidroside compound of the present invention according to the method described in the document, and then carried out carbon spectrum and hydrogen spectrum analysis, but could not obtain the carbon spectrum and hydrogen spectrum data of the acetylated product and its supplementary document. Therefore, the hydroxy- ⁇ -salidroside compound obtained by the present invention is different from the compound obtained in the document.
  • the feed amount was simultaneously increased by 10 times, the reaction was stirred at 35° C., and the reaction was carried out for 24 hours to obtain an enzyme catalytic reaction solution.
  • FIG. 1 The TLC analysis spectrum of the reaction solution of Example 1 is shown in Figure 1, which shows that new substances are produced in the enzyme-catalyzed reaction solution.
  • Figure 1 shows that new substances are produced in the enzyme-catalyzed reaction solution.
  • lane 1 is ⁇ -salidroside
  • lane 2 is the new substance produced by the enzyme-catalyzed reaction.
  • Example 1 According to the above chromatographic conditions, the purified reaction solution of Example 1 was subjected to HPLC analysis. The HPLC spectrum is shown in FIG2 .
  • the LC-MS spectrum is shown in FIG3 .
  • a semi-preparative HPLC was used (BRIX 2850, Beijing Chengda Instrument Co., Ltd.).
  • the chromatographic column was COSMOSIL 5C18-MS-II (250*20mm).
  • the mobile phase was acetonitrile: 0.1% formic acid aqueous solution (v/v) with an isocratic elution ratio of 1:9.
  • a photodiode array detector was used with a detection wavelength of UV 280nm.
  • the injection volume was 100 ⁇ L and the flow rate was 18mL/min.
  • the H-NMR spectrum of the product in Example 1 has two single peaks (s, 1H: OH) at 8.64ppm and 8.69ppm, which is the "characteristic of bisphenol hydroxyl".
  • ⁇ -Salidroside is a single peak at 9.10ppm, which is the "characteristic of monophenol hydroxyl”
  • the C-NMR spectrum of the product in Example 1 has 14 characteristic peaks, while ⁇ -Salidroside has only 12 characteristic peaks, which are obviously different.
  • the HPLC spectrum, LC-MS spectrum, H-NMR spectrum, and C-NMR spectrum of the product of the enzyme-catalyzed reaction of Example 2-4 are the same as those of the reaction solution of Example 1. Therefore, the substance obtained by the enzyme-catalyzed reaction of Example 2-4 is also hydroxy- ⁇ -salidroside.
  • Example 6-8 Based on Example 3, an ammonium salt, such as ammonium chloride, ammonium acetate, or ammonium sulfate, is added to the reaction system to carry out an enzymatic reaction.
  • an ammonium salt such as ammonium chloride, ammonium acetate, or ammonium sulfate.
  • DPPH free radical scavenging experiment Free radicals directly or indirectly play a strong oxidative role and are widely involved in the physiological and pathological processes of the body. When there are excessive free radicals in the body, they can damage the body through oxidation. Salvianolic acid compounds are donors of phenolic hydroxyl groups and have a structural basis for antioxidant activity. This experiment uses the DPPH free radical scavenging reaction to study the scavenging efficiency of hydroxy- ⁇ -salidroside, ⁇ -salidroside, and ⁇ -salidroside on DPPH free radicals.
  • the DPPH free radical scavenging rate was calculated according to the following formula. The results are shown in Table 4.
  • Enzyme reaction tube C Take 1 mL of sodium hydrogen phosphate-citrate buffer, add 0.5 mL of tyrosinase, mix well, incubate in a 37°C water bath for 10 min, then add 2 mL of L-DOPA, control the reaction time to 5 min, and immediately measure the absorbance at 475 nm.
  • Solvent background C 0 Take 1.5 mL of sodium hydrogen phosphate-citrate buffer, incubate in a 37°C water bath for 10 min, add 2 mL of L-DOPA, control the reaction time to 5 min, and immediately measure the absorbance at 475 nm.
  • Hydroxy- ⁇ -salidroside sample solution The target substance of Example 4 was prepared according to the method described in Example 5, and prepared into a 10 mg/mL solution.
  • ⁇ -Salidroside sample solution The ⁇ -Salidroside is the raw material for preparing hydroxy- ⁇ -Salidroside in the examples, and is prepared into a 10 mg/mL solution.
  • the drugs were administered according to the following schedule starting from the second day after the 7th week of modeling and continued for 30 days.
  • AD model group 1 Each rat was intragastrically administered with hydroxy- ⁇ -salidroside sample solution once a day, with a dose of 5 mg per kg body weight.
  • AD model group 3 Each rat was intragastrically administered with ⁇ -salidroside sample solution once a day, with a dose of 5 mg per kg body weight.
  • AD model group 2 and blank control group each rat was intragastrically administered with the sample solution once a day, with the same volume of distilled water as that of the AD model group 1.
  • the constant temperature swimming pool used in the experiment was 120 cm in diameter and 50 cm in height, with a water depth of 30 cm and a water temperature maintained at (22 ⁇ 1)°C.
  • Two objects were randomly hung on the wall of the pool as close visual cues.
  • the built-in platform was 7 cm in diameter and was submerged 2 cm under water. There were several small holes on it to provide a surface for rats to stand easily.
  • the environment information was kept fixed throughout the experiment. Milk was added to the pool to make the platform invisible. The maze was divided into 4 quadrants, and the platform was placed in the center of one of the quadrants as the only way for the animals to escape the water environment in the experiment.
  • the platform was placed in a quadrant, and the positioning navigation test was performed, and the escape latency was recorded.
  • the test was repeated for 3 consecutive days, 3 times a day, and the entry point was changed each time.
  • the order of the entry points for all rats was kept consistent every day (clockwise).
  • the rats were gently placed in the water toward the pool wall. 120s was set as the longest escape latency, and the recording stopped automatically after 120s. If the rat found the platform within 120s, its actual escape latency was recorded; if the platform was not found within 120s, the experimenter led it to the platform and stayed for 10s, and the escape latency was recorded as 120s.
  • the Morris water maze test is an experiment that forces experimental animals (rats and mice) to swim and learn to find a platform hidden in the water. It is mainly used to test the learning and memory ability of experimental animals for spatial position and direction (spatial positioning).
  • Escape latency refers to the time it takes for an animal to swim from the starting position in the water maze to find the hidden platform. As the number of experiments increases, the time it takes for the animal to find the platform should gradually decrease, indicating that they have made progress in learning and remembering the location of the platform. A longer escape latency may indicate that the animal's spatial learning and memory abilities are impaired.
  • the AD model group 1 was the hydroxy- ⁇ -salidroside treatment group, and its escape latency was shorter than that of the AD model group 3 ( ⁇ -salidroside treatment group) and the AD model group 2 (blank model group), indicating that the AD model rats in the AD3 group needed a shorter time to find the underwater hidden platform, and that hydroxy- ⁇ -salidroside had a certain effect on improving the spatial learning and memory ability of the AD model rats.

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Abstract

本发明涉及一种红景天苷的衍生物，具体涉及一种羟基-α-红景天苷化合物，所述羟基-α-红景天苷化合物，在清除DPPH自由基、美白方面以及在改善学习记忆障碍方面优于α-红景天苷，可作为新的功能性成分添加到化妆品、功能食品或药品组合物中。本发明所述羟基-α-红景天苷的制备方法，以α-红景天苷为底物，在羟化酶的作用下，合成羟基-α-红景天苷。

Description

红景天苷的衍生物及制备方法和应用
相关申请的交叉引用
本发明要求于2023年6月8日提交的标题为“一种红景天苷的衍生物及制备方法和应用”的中国专利申请CN202310670529.3的优先权，并且要求于2024年2月24日提交的标题为“红景天苷的衍生物及制备方法和应用”的中国专利申请CN202410159803.5的优先权。上述申请的全部内容通过引用并入本文，用于所有目的。
技术领域
本发明涉及一种红景天苷的衍生物，具体涉及一种羟基-α-红景天苷化合物。
背景技术
公开该背景技术部分的信息仅仅旨在增加对本发明的总体背景的理解，而不必然被视为承认或以任何形式暗示该信息构成已经成为本领域一般技术人员所公知的现有技术。
红景天是一味珍贵中药，2020版《中国药典》收录为景天科大花红景天的干燥根和根茎，产于中国西藏等地，生长于海拔4050米至5400米左右的地区。由红景天制备的红景天片、心脑欣胶囊等药品具有活血化瘀，通脉止痛的功效，红景天胶囊等保健食品的功能为增强免疫力、缓解体力疲劳。红景天苷是中药红景天的有效活性成分，据报道，红景天苷具有保护心脑血管、抗疲劳、抗抑郁、抗衰老、抗缺氧、抗辐射、抗肿瘤、免疫调节、美白祛斑等生理活性。天然状态下的红景天苷(植物来源红景天苷)为β-红景天苷，其糖苷键为β型。
中国专利202211359878.5，公开了一种α-红景天苷及其制备方法与应用，所述α-红景天苷，化学名称为：2-(4-羟基苯基)乙基-α-D-葡萄糖苷，分子量为300.23，化学式如下，其具有优异的清除自由基活性，可作为化妆品、抗疲劳保健品等产品的主要原料。
发明内容
发明目的：处于对产品迭代的需求，申请人对红景天苷及其衍生物进行了研究，获得了羟基-α-红景天苷。另外，本发明还提供了羟基-α-红景天苷的制备方法及应用。
现有技术没有关于羟基-α-红景天苷的报道。
技术方案：
第一方面，本发明提供了一种羟基-α-红景天苷化合物，化学名称：2-(3,4-二羟基苯基)乙基-α-D-葡萄糖苷，化学式见式I，其在羟基酪醇的醇羟基上具有α-葡萄糖苷键。
本发明所述羟基-α-红景天苷经LC-MS分析，其分子量为316.108。
本发明所述羟基-α-红景天苷的H-NMR谱：1HNMR(600MHz,DMSO):δ2.64-2.72(m,2H),3.02-3.08(m,1H),3.17-3.21(m,1H),3.31-3.37(m,4H),3.37-3.46(m,2H),3.46-3.51(m,1H),3.57-3.63(m,1H),3.67-3.73(m,1H),4.44(t,1H),4.57(d,1H),4.68(d,J＝3.6Hz,1H:β-H),4.73(d,1H),4.83(d,1H),8.64(s,1H:OH),8.69(s,1H:OH)。
所述羟基-α-红景天苷的C-NMR谱：13C NMR(151MHz,DMSO):δ35.5,61.4,68.9,70.8,72.5,73.3,73.8,99.1,115.9,116.7,120.0,129.9,144.0,145.5。
第二方面，本发明提供了所述羟基-α-红景天苷化合物的制备方法。
所述制备方法，包括将α-红景天苷溶解于缓冲溶液中，再加入羟化酶，在反应体系中进行羟化反应制备得到。
优选的，所述羟化酶可以选择市售商品化羟化酶(Hydroxylase)制剂。
进一步优选的，所述市售商品化羟化酶酶制剂来源于默克有限公司、上海麦克林生化科技股份有限公司和上海源叶生物科技有限公司，包括但不限于单酚单加氧酶(Monophenol monooxygenase)、多酚氧化酶(Polyphenol Oxidase)、酪氨酸酶(Tyrosinase)等。
优选的，所述反应体系中还可以加入不影响酶促反应的表面活性剂，如聚山梨酯(Tween-20)或失水山梨醇脂肪酸酯(Span)。
本发明中，所述羟基-α-红景天苷的制备也可以在有机溶剂中进行，反应中使用的有机溶剂只要是不影响反应的种类和浓度范围即可，具体包括DMSO、2-丙醇、乙醇中的一种溶剂或两种以上的混合溶剂。
优选的，所述反应体系中，含有铵盐。
所述铵盐，优选的，包括但不限于氯化铵、乙酸铵、或硫酸铵。
铵盐能够提高底物α-红景天苷的转化率。
第三方面，本发明提供了所述羟基-α-红景天苷化合物在制备具有清除自由基功能的产品中的应用；或具有美白功能的化妆品方面的应用。
优选的，所述自由基为1,1-二苯基-2-三硝基苯肼(DPPH)自由基。
所述应用，优选的，所述产品为具有清除自由基功能的化妆品。
优选的，所述清除自由基功能的产品为功能食品。
本发明进一步提供了一种组合物，含有羟基-α-红景天苷。
优选的，所述组合物具有清除自由基的功能，或美白肌肤的功能。
优选的，所述自由基为DPPH自由基。
所述含有羟基-α-红景天苷的化妆品组合物，其品类选自水剂类、油剂类、乳剂类、凝胶类产品、膏状产品，或其他，可以按现有化妆品技术规范生产。
含有羟基-α-红景天苷的功能食品组合物，其剂型选自胶囊剂、片剂、膏剂、水剂，或其他，可以按现有功能食品技术规范生产。
第四方面，本发明还提供了所述羟基-α-红景天苷化合物在制备改善学习记忆障碍药品中的应用。
优选的，所述改善学习记忆障碍药品包括但不限于治疗阿尔兹海默症的药品。
优选的，所述改善学习记忆障碍药品包括但不限于治疗帕金森症的药品。
学习记忆障碍是许多疾病可能出现的症状群。引起学习记忆障碍的原因多种多样，如阿尔茨海默病和帕金森病等在内的神经退行性病变、应激、衰老、脑缺血性损伤、脑外伤、高血压、糖尿病、更年期综合症和抑郁症等。其中阿尔茨海默病是最常见的一种学习记忆障碍性疾病，临床上的药物只能减轻症状，不能延缓或终止其进程，并且在不同患者身上产生的疗效不一致，有较多的不良反应。另外，目前人们的工作和生活节奏加快，竞争压力增加，长期处于慢性应激状态中，导致机体协调失衡、功能紊乱，尤其是影响大脑认知功能，导致学习记忆能力下降等。因此，现在迫切需要改善学习记忆障碍的药物。研究发现本发明所述羟基-α-红景天苷具有改善模型鼠的学习记忆水平，表现在可显著提高大鼠对新位置物体的相对辨别指数，缩短水迷宫实验中的寻台潜伏期，提高穿台次数，降低避暗实验中的错误次数和暗室时间，表明羟基-α-红景天苷能够较好地改善学习记忆障碍。
第五方面，提供了一种清除自由基的方法，包括向目标区域或受试者施用有效量的羟基-α-红景天苷或包含有效量的羟基-α-红景天苷的组合物。所述自由基为DPPH自由基。
第六方面，提供了一种美白皮肤的方法，包括向皮肤施用有效量的羟基-α-红景天苷或包含有效量的羟基-α-红景天苷的组合物。
第七方面，提供了一种改善学习记忆障碍的方法，包括向受试者施用有效量的羟基-α-红景天苷或包含有效量的羟基-α-红景天苷的组合物。
本发明所述“受试者”指任何需要治疗、预防或诊断的生物体，优选为哺乳动物，更优选为人类。
本发明所述“有效量”指足以实现所需治疗、预防或诊断效果的化合物的量。该量可以根据受试者的年龄、体重、健康状况、性别以及具体的治疗、预防或诊断目的进行调整。有效量可通过常规药理学测试方法、体外试验、动物模型试验或临床试验测得。通常，所述有效量在约0.01mg/kg至1000mg/kg体重范围内，更优选在约0.1mg/kg至100mg/kg体重范围内。
有益效果：
本发明提供了一种红景天苷衍生物，即羟基-α-红景天苷，与α-红景天苷相比，在清除DPPH自由基方面，具有更高的活性，可作为新的功能性成分添加入化妆品中或保健品中；所述羟基-α-红景天苷还具有改善模型鼠的学习记忆水平的优异性能，可作为制备阿尔兹海默症药物、或帕金森症药物的开发应用。
附图说明
构成本发明的一部分的说明书附图用来提供对本发明的进一步理解，本发明的示意性实施例及其说明用于解释本发明，并不构成对本发明的不当限定。以下，结合附图来详细说明本发明的实施方案，其中：
图1：实施例1酶催化反应液的TLC谱图；图中，从左至右分别为泳道1、泳道2；泳道1为α-红景天苷，泳道2为羟基-α-红景天苷。
图2：实施例1酶催化反应液的HPLC谱图。
图3：实施例1酶催化反应液HPLC谱图中，rt＝6.97min物质的LC-MS谱图。
图4：实施例1酶催化反应液HPLC谱图中，rt＝6.97min物质的H-NMR谱图。
图5：实施例1酶催化反应液HPLC谱图中，rt＝6.97min物质的C-NMR谱图。
具体实施方式
下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件或按照制造厂商所建议的条件。
除非另行定义，文中所使用的所有专业与科学用语与本领域熟练人员所熟悉的意义相同。本发明所使用的试剂或原料均可通过常规途径购买获得，如无特殊说明，本发明所使用的试剂或原料均按照本领域常规方式使用或者按照产品说明书使用。此外，任何与所记载内容相似或均等的方法及材料皆可应用于本发明方法中。文中所述的较佳实施方法与材料仅作示范之用。
本发明所述羟基-α-红景天苷，化学名称为：2-(3,4-二羟基苯基)乙基-α-D-葡萄糖苷，结构式见式I。
本发明所述羟基-α-红景天苷的制备方法，以α-红景天苷为底物，在羟化酶的作用下实现羟基化。
所述羟化酶，可以选择市售商品化羟化酶酶制剂，来源于默克有限公司、上海麦克林生化科技股份有限公司和上海源叶生物科技有限公司，包括儿茶酚氧化酶但不限于单酚单加氧酶、多酚氧化酶、酪氨酸酶等。
羟化酶酶制剂的使用方法包括以粉末或液体的形式加入反应体系中，也可将酶制剂固定在树脂上制成固定化酶制剂加入到反应体系中，固定化酶制剂可以重复使用。
本发明所述羟基-α-红景天苷的制备方法，是将羟化酶加入α-红景天苷磷酸缓冲液中，进行羟化酶反应。酶反应溶液中可加入不影响酶促反应的表面活性剂，例如Tween-20和Span等试剂。
在使用上述羟化酶制备羟基-α-红景天苷化合物时，羟化酶的用量和反应条件对生产效率有很大的影响，因此选择合适的酶量和反应时间等反应条件至关重要。
反应可以在溶剂存在的情况下进行，反应中使用的溶剂只要是不影响反应的种类和浓度范围即可，具体可包括DMSO、2-丙醇、乙醇等常用试剂。这些溶剂可以单独使用，也可以同时使用两种或两种以上。
通过加热或改变pH值等方法可以导致本发明使用的酶失活，从而停止酶催化反应。
上述含羟基-α-红景天苷化合物的混合物、混合物的水解产物、混合物的酶灭活产物、混合物的纯化产物，其干燥的粉末产品可用于含羟基-α-红景天苷化合物的食品、饮料和化妆品中，也可作为食品、特医食品、保健品或药品的成分，用于食品和饮料、化妆品、特医食品、保健品或药品的生产。
TLC分析条件：采用薄层色谱法对酶反应溶液进行快速分析，具体分析条件如下：使用Merck公司生产的TLC板(Merck Kieselgel 60F254)，以氯仿:甲醇:乙酸＝5:1:1的溶液为展开溶剂。
HPLC分析条件：高效液相色谱仪：日立HPLC 5440色谱仪，色谱柱：Kromasil 100-5C18(250*4.6mm)，检测器：光电二极管阵列(DAD 280nm)，检测波长：UV 280nm，进样量：10μL，流速：1mL/min，柱温：30℃，流动相：乙腈：0.1％甲酸水溶液(v/v)等度洗脱。
LC-MS分析条件：色谱柱：Kromasil 100-5C18(250*4.6mm)，检测器：光电二极管阵列(DAD 280nm)，检测波长：280nm，进样量：10μL，流速：1mL/min，柱温：30℃，流动相同上检测时间为5-15min的流动相；H-ESI模式，分子量扫描范围50～800。
文献Enzymatic routes for the production of mono-and di-glucosylated derivatives of hydroxytyrosol”,Antonio Trincone et al.《Bioresource Technology》》第115卷是一篇关于“酶法生产羟基酪醇的单葡糖基化和二葡糖基化衍生物”的文章，通过将来自Aplysia.fasciata的内脏物质匀浆加入到1.5mL在50mM乙酸钾缓冲液中的10mM受体溶液中(pH 5.5，含有150mM麦芽糖)，获得了(3,4-二羟基苯基)乙基-α-D-吡喃葡糖苷化合物，并通过对反应所得产品进行乙酰化所得核磁数据推断获得了目标化合物。但该文献缺少质谱鉴定数据，没有公开具体化合物的分子量。也没有作为α-葡萄糖苷关键氢原子的化学位移数据，没有公开具体化合物的化学结构式，不能确认所述化合物的立体构型。
本发明的发明人根据该文献描述的方法，对本发明所述羟基-α-红景天苷化合物进行乙酰化处理，然后进行碳谱和氢谱进行分析，不能获得所述乙酰化产品及其补充文献所述碳谱和氢谱数据，因此本发明所得羟基-α-红景天苷化合物与文献所得化合物不同。
实施例1.酪氨酸酶羟基化实验
取α-红景天苷(分子量300.23)500mg，加入50mL 0.2M磷酸盐缓冲液(并调整pH至10)，加入500mg抗坏血酸，再加入3mg酪氨酸酶(T828256-100KU，麦克林)，加水补足体积至100mL，25℃搅拌反应，反应24小时，得酶催化反应溶液。
实施例2.多酚氧化酶羟基化实验
取α-红景天苷(分子量300.23)100mg，加入50mL 0.2M磷酸盐缓冲液(并调整pH至10)，加入100mg抗坏血酸，再加入3mg酪氨酸酶(S10171-100KU，上海源叶生物科技有限公司)，加水补足体积至100mL，15℃搅拌反应，反应50小时，得酶催化反应溶液。
实施例3.单酚单加氧酶羟基化实验
取α-红景天苷(分子量300.23)500mg，加入50mL 0.2M磷酸盐缓冲液(pH9)，加入500mg抗坏血酸，再加入1.5g单酚单加氧酶(T3824-50KU，Merck)，加水补足体积至100mL，35℃搅拌反应，反应24小时，得酶催化反应溶液。
实施例4.单酚单加氧酶羟基化实验
依实施例3所述方法，投料量同步放大10倍，35℃搅拌反应，反应24小时，得酶催化反应溶液。
实施例5.羟基-α-红景天苷的纯化及结构分析
对实施例1-4反应液进行TLC、HPLC、LC-MS、NMR分析。
(1)TLC分析
实施例1反应液TLC分析图谱见图1，显示酶催化反应液中有新物质产生。图中，从左至右分别为为泳道1、泳道2；泳道1为α-红景天苷，泳道2为酶催化反应产生的新物质。
(2)HPLC分析
依照上述色谱条件，对实施例1纯化后反应液进行HPLC分析，HPLC图谱详见图2。
图2所示HPLC图谱显示，实施例1纯化后反应液主要有两个峰，出峰时间分别是rt＝6.97min和rt＝9.98min，其中rt＝9.98min处的峰是α-红景天苷，rt＝6.97min的峰与α-红景天苷出峰时间有明显的差异。
(3)LC-MS分析
对实施例1反应液HPLC图谱中rt＝6.97min的物质进行LC-MS分析，得到分子离子峰[M-1]的测量值为315.108，LC-MS图谱见图3。
根据光电二极管阵列检测器显示的紫外光谱图和LC-MS图谱，初步分析HPLC图谱中保留时间rt＝6.97min的新峰物质的分子量为316.108，可能是α-红景天苷羟基化形成的糖苷，可能是羟基-α-红景天苷。
(4)分离纯化
采用HPLC法分离纯化实施例1-4得到的酶催化反应液，并收集rt＝6.97min的物质，其中，HPLC法的设备和条件包括：
半制备型高效液相色谱仪(北京橙达仪器有限公司BRIX 2850)，色谱柱为COSMOSIL 5C18-MS-II(250*20mm)，流动相为乙腈:0.1％甲酸水溶液(v/v)以1:9的比例等度洗脱，使用光电二极管阵列检测器，检测波长为UV 280nm，进样量为100μL，流速为18mL/min。
将收集得到的分别来自实施例1-4的rt＝6.97min的物质，分别进行浓缩冻干，得到冻干固体，作为实施例1-4的目标物质，用于后续实验。
(5)H-NMR和C-NMR谱图分析
取上述冻干样品进行H-NMR、C-NMR分析。
实施例1反应液HPLC图谱中rt＝6.97min的物质的H-NMR谱图见图4，C-NMR谱图见图5。
1H-NMR(600MHz,DMSO):δ2.64-2.72(m,2H),3.02-3.08(m,1H),3.17-3.21(m,1H),3.31-3.37(m,4H),3.37-3.46(m,2H),3.46-3.51(m,1H),3.57-3.63(m,1H),3.67-3.73(m,1H),4.44(t,1H),4.57(d,1H),4.68(d,J＝3.6Hz,1H:β-H),4.73(d,1H),4.83(d,1H),8.64(s,1H:OH),8.69(s,1H:OH)。
13C-NMR(151MHz,DMSO):δ35.5,61.4,68.9,70.8,72.5,73.3,73.8,99.1,115.9,116.7,120.0,129.9,144.0,145.5。
依据实施例1产品的H-NMR谱和C-NMR谱，结合中国专利申请202211359878.5所公开的α-红景天苷的H-NMR谱和C-NMR谱，实施例1产品的H-NMR谱在8.64ppm和8.69ppm处，存在两个单峰(s,1H:OH)，这是“双酚羟基的特征”。α-红景天苷在9.10ppm处是单峰，这是“单酚羟基的特征”，且实施例1产品的C-NMR谱出现14个特征峰，而α-红景天苷只存在12个特征峰，二者明显不同。
综上，从TLC图谱和HPLC图谱可见，酶催化反应产生了新物质；从LC-MS分析，新物质分子量为316.108；再结合H-NMR和C-NMR图谱，证实实施例1酶催化反应所得物质为羟基-α-红景天苷。
实施例2-4酶催化反应产物的HPLC图谱、LC-MS图谱、H-NMR、C-NMR图谱同实施例1反应液。因此，实施例2-4酶催化反应所得物质也为羟基-α-红景天苷。
HPLC法测定实施例1-4反应液中的羟基-α-红景天苷浓度，结果记录于表1。
表1
实施例6-8.在实施例3的基础上，向反应体系中添加铵盐，如氯化铵、或乙酸铵、或硫酸铵，进行酶促反应。具体如下：
分别取α-红景天苷(分子量300.23)500mg，加入50mL0.2M磷酸盐缓冲液(pH9)，加入500mg抗坏血酸，分别添加10mL的0.2M铵盐(具体铵盐品种见表2)，再加入1.5g单酚单加氧酶(T3824-50KU，Merck)，加水补足体积至100mL，35℃搅拌反应，反应24小时，得酶催化反应溶液，HPLC法检测反应液中羟基-α-红景天苷，结果记录于表2。
表2

表2数据说明，氯化铵、乙酸铵均能够提高酶促反应中底物α-红景天苷的转化率。
实施例9.自由基清除实验，所用物料与试剂如见表3。
表3.物料与试剂
DPPH自由基的清除实验：自由基直接或间接地发挥强氧化作用，广泛地参与机体的生理与病理过程。机体自由基过量时，能通过氧化作用损伤机体。丹酚酸类化合物是酚羟基的供体，具有抗氧化活性的结构基础。本实验采用DPPH自由基清除反应，研究羟基-α-红景天苷、α-红景天苷、β-红景天苷对DPPH自由基的清除效率。
溶液制备：
(1)羟基-α-红景天苷溶液制备：按照实施例5的方法分别制备实施例1-3的目标物质，并按常规方法分别制备为500mg/L水溶液。
(2)α-红景天苷溶液制备：按常规方法制备为500mg/L水溶液。
(3)β-红景天苷溶液制备：按常规方法制备为500mg/L水溶液。
(4)DPPH乙醇溶液制备：按常规方法制备为0.04mg/mL的无水乙醇溶液。
2.DPPH自由基清除实验：
(1)分别取羟基-α-红景天苷、α-红景天苷、β-红景天苷水溶液各0.9mL，分别加入DPPH无水乙醇溶液0.9mL，混合均匀，置于暗处静置，分别于0.5h、1h、2h、3h、4h、5h取样，于517nm波长处测定吸光度，计算三个平行的平均值，记为Ai。
(2)分别取羟基-α-红景天苷、α-红景天苷、β-红景天苷水溶液各0.9mL，分别加入无水乙醇0.9mL，混合均匀，置于暗处静置，分别于0.5h、1h、2h、3h、4h、5h取样，于517nm波长处测定吸光度，计算三个平行的平均值，记为Aj。
(3)取蒸馏水0.9mL，加入DPPH乙醇溶液0.9mL，混合均匀，于517nm波长处测定吸光度，计算三个平行的平均值，记为A4。
(4)0.9mL无水乙醇，加蒸馏水0.9mL，混合均匀，作为空白。
依据如下公式计算DPPH自由基清除率，结果见表4。
表4.DPPH自由基清除率
表4数据说明：羟基α-红景天苷清除自由基的能力0.5小时即达最高值，为α-红景天苷和β-红景天苷的4倍以上。
实施例10.酪氨酸酶活性抑制实验
物料与试剂如见表5。
表5.物料与试剂
仪器：752N紫外可见分光光度计，上海佑科仪器仪表有限公司。
溶液配制：
(1)α-红景天苷溶液配制：按常规方法配制成1mg/mL的水溶液。
(2)羟基-α-红景天苷溶液制备：按照实施例5的方法分别制备实施例1-3的目标物质，并按常规方法分别配制成1mg/mL的水溶液。
(3)磷酸氢二钠-柠檬酸缓冲液配制：
1)配制0.2mol/L的磷酸氢二钠和0.1mol/L的柠檬酸水溶液。
2)取磷酸氢二钠水溶液154.5mL，柠檬酸水溶液45.5mL，混匀，调整pH为6.8(pH计测量，可用0.2mol/L的磷酸氢二钠或0.1mol/L的柠檬酸调整pH值)。
(4)酪氨酸酶溶液配制：用上述磷酸氢二钠-柠檬酸缓冲液制备100u/mL的酪氨酸酶溶液。
(5)左旋多巴溶液配制：用上述磷酸氢二钠-柠檬酸缓冲液制备1mg/mL左旋多巴溶液。
实验方案：
(1)分别取羟基-α-红景天苷溶液、α-红景天苷溶液各1mL，分别加入酪氨酸酶0.5mL，混匀，37℃水浴10min，再分别加入左旋多巴2mL，控制反应时间5min，立刻分别于475nm处测定吸光度T。
(2)样品本底T0：分别取羟基-α-红景天苷溶液、α-红景天苷各1mL，分别加入磷酸氢二钠-柠檬酸缓冲液0.5mL，混匀，37℃水浴10min，再加入左旋多巴2mL，控制反应时间5min，立刻于475nm处测定吸光度。
(3)酶反应管C：取磷酸氢二钠-柠檬酸缓冲液1mL，加入酪氨酸酶0.5mL，混匀，37℃水浴10min，再加入左旋多巴2mL，控制反应时间5min，立刻于475nm处测定吸光度。
(4)溶剂本底C0：取磷酸氢二钠-柠檬酸缓冲液1.5mL，37℃水浴10min，加入左旋多巴2mL，控制反应时间5min，立刻于475nm处测定吸光度。
依据以下公式计算酪氨酸酶活性的抑制率，结果记录于表6。
表6
表6数据说明：实施例1、实施例3所得羟基-α-红景天苷溶液的酪氨酸酶活性抑制率为α-红景天苷的46倍或以上。
实施例11.羟基-α-红景天苷对Wistar大鼠学习记忆影响实验研究
实验动物：Wistar雄性青年大鼠，体重290-315g，56只。随机分为4组，每组14只，随机分为空白对照组、AD模型1组、AD模型2组和AD模型3组。
羟基-α-红景天苷样品溶液：按实施例5中所述方法制备实施例4的目标物质，并制备为10mg/mL溶液。
α-红景天苷样品溶液：所述α-红景天苷为实施例中制备羟基-α-红景天苷的原料，配制为10mg/mL溶液。
实验方法
造模：Wistar雄性大鼠AD模型的建立
AD模型1组-3组造模方法:每日分别给Wistar雄性大鼠腹腔注射D-半乳糖120mg/kg和亚硝酸钠80mg/kg，连续7周完成造模；空白对照组腹腔注射等体积生理盐水。
造模第7周结束后第2天开始按如下方案给药，连续给药30天。
AD模型1组：各大鼠每天灌胃羟基-α-红景天苷样品溶液1次，剂量按每kg体重给药5mg计算。
AD模型3组：各大鼠每天灌胃α-红景天苷样品溶液1次，剂量按每kg体重给药5mg计算。
AD模型2组、空白对照组：各大鼠每天灌胃样品溶液一次，剂量为与AD模型1组相同体积的蒸馏水。
1.水迷宫实验
第31天开始，进行Morris水迷宫试验，试验期间继续按如上方案给药。
试验用恒温游泳池直径为120cm、高50cm，池中水深30cm，水温维持在(22±1)℃，池壁上随意挂两个物体作为近距离视觉暗示。内置平台直径7cm，平台没于水下2cm，上面有几个小孔以提供一个大鼠容易站稳的表面。
整个实验周期内保持环境信息固定不变。水池中加入牛奶，使平台不可见。迷宫被均分为4个象限，将平台置于其中一个象限中央，作为实验中动物逃离水环境的唯一途径。
Morris水迷宫试验的第1天将大鼠放入水池中(不放入平台)自由游泳2min，使其熟悉迷宫内的环境。
Morris水迷宫试验第2天起，将平台置于某一象限，进行定位航行试验，记录逃逸潜伏期，连续试验3天，每天3次，每次变换入水点位置，且每天所有大鼠入水点的顺序保持一致(顺时针)。试验期间，应注意保持平台位置固定不变，选择未放平台的其他3个象限作为大鼠的入水点，将大鼠向池壁轻放入水中。设定120s为最长逃避潜伏期，120s后自动停止记录。如果大鼠在120s内找到平台，记录其实际逃避潜伏期；如果在120s未找到平台，由实验者将其引上平台并停留10s，逃避潜伏期记为120s。
记录各组大鼠的水迷宫逃避潜伏期，每组每日逃避潜伏期平均值记录于表7。
表7大鼠水迷宫逃避潜伏期测定结果
Morris水迷宫试验是一种强迫实验动物(大鼠、小鼠)游泳，学习寻找隐藏在水中平台的一种实验，主要用于测试实验动物对空间位置觉和方向觉(空间定位)的学习记忆能力。
逃逸潜伏期(Escape Latency)：逃逸潜伏期是指动物从水迷宫中的起始位置游动到找到隐藏平台所需的时间。随着实验次数的增加，动物找到平台的时间应逐渐缩短，这表明它们在学习和记忆平台位置方面取得了进步。较长的逃逸潜伏期可能表示动物的空间学习和记忆能力受到损害。
表7数据说明：
(1)在定位航行实验的第2-4天，AD1-3组模型大鼠的逃逸潜伏期较空白对照组大鼠更长(P＜0.05)，说明AD1-3组模型大鼠找到水下隐藏平台需要更长的时间。
(2)AD模型1组为羟基-α-红景天苷用药组，其逃逸潜伏期较AD模型3组(α-红景天苷用药组)、AD模型2组更短(空白模型组)，说明AD3组模型大鼠找到水下隐藏平台需要更短的时间，以及羟基-α-红景天苷对AD模型大鼠有一定的提高空间学习记忆能力。
应注意的是，以上实例仅用于说明本发明的技术方案而非对其进行限制。尽管参照所给出的实例对本发明进行了详细说明，但是本领域的普通技术人员可根据需要对本发明的技术方案进行修改或者等同替换，而不脱离本发明技术方案的精神和范围。

Claims (10)

  1. 一种羟基-α-红景天苷，化学名称为：2-(3,4-二羟基苯基)乙基-α-D-葡萄糖苷，其结构式如式I，
  2. 权利要求书1所述羟基-α-红景天苷在制备具有清除自由基功能的产品中的应用；
    优选的，所述自由基为DPPH自由基，所述产品为化妆品或功能食品。
  3. 权利要求书1所述羟基-α-红景天苷在制备具有美白功能的产品中的应用。
  4. 权利要求1所述羟基-α-红景天苷在制备改善学习记忆障碍的药品中的应用；
    优选的，所述改善学习记忆障碍的药品包括但不限于治疗阿尔兹海默症的药品；
    优选的，所述改善学习记忆障碍的药品包括但不限于治疗帕金森症的药品。
  5. 一种组合物，其特征在于，含有权利要求1所述的羟基-α-红景天苷。
  6. 如权利要求5所述的组合物在清除自由基方面的应用。
  7. 如权利要求5所述的组合物在制备美白化妆品方面的应用。
  8. 权利要求5所述的组合物在制备改善学习记忆障碍的药品中的应用；
    优选的，所述改善学习记忆障碍的药品包括但不限于治疗阿尔兹海默症的药品；
    优选的，所述改善学习记忆障碍的药品包括但不限于治疗帕金森症的药品。
  9. 权利要求1所述羟基-α-红景天苷的制备方法，其特征在于，在含有α-红景天苷的溶液中，加入羟化酶进行反应；
    优选的，所述羟化酶选自单酚单加氧酶、多酚氧化酶或酪氨酸酶。
  10. 如权利要求9所述的制备方法，其特征在于，所述反应中还加入铵盐；
    优选的，所述铵盐为氯化铵、乙酸铵或硫酸铵。
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