JP2019521076A - Composition for Improving cognitive function comprising 3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflavone - Google Patents

Abstract

In one aspect, the present invention relates to a composition for improving cognitive function lowering comprising a post-fermented tea ketone fraction as an active ingredient. In another aspect, the present invention relates to 3-O-galloyl- separated from a post-fermented tea ketone fraction. 3,3 ′, 5,5 ′, 7-pentahydroxyflavone (3-O-galloyl-3,3 ′, 5,5 ′, 7-pentahydroxyflavan), its isomers, pharmaceutically acceptable salts thereof, With regard to the cognitive function improvement composition containing the prodrug, the hydrate, or the solvate as an active ingredient, it is possible to effectively suppress the aggregation of beta amyloid using one aspect of the present invention, Since BDNF expression and DNMT1 expression can be regulated, the present invention can be utilized in various fields related to cognitive decline in one aspect. [Selection] Figure 5

Description

  This specification is a cognition including 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavone (3-O-garyl-3,3 ', 5,5', 7-pentahydroxyflavan) The present invention relates to a composition for improving function.

  With the improvement of living standards and the development of the medical industry, it has become possible to treat cancer and other intractable diseases, and the human life has been prolonged, but the aging of the population accompanying it has resulted in an increase in late-onset degenerative neurological diseases. This is rather a relative decline in the quality of life. Neuronal dysfunction and damage are said to be induced by aggregation proteins, and many neurological diseases are characterized by such conditions. Such neurological diseases include diseases such as Alzheimer's disease.

  Alzheimer's disease is a progressive nerve that shows symptoms such as cognitive decline, memory decline, irreversible memory loss, loss of orientation, loss of cognitive ability, depression, amnesia and language impairment while damage to brain neurons. It is a degenerative disease. Alzheimer's disease is the third leading cause of death associated with aging following cerebrovascular disease, cancer, and when it occurs, the average duration of disease is more than 10 years, so the social and mental burden of the parents Burden is increasing. The most representative cause of Alzheimer's disease known to date includes abnormal aggregation of beta-amyloid (β-amyloid, also referred to as Aβ, amyloid beta). That is, beta-amyloid, which is normally present in the brain, abnormally aggregates to form amyloid plaques (Aβ plaque), thereby causing damage to neural cells and neural cell connections and causing disease.

  With the growth of the elderly population, the need for treatment and prevention of aging, degenerative neurological diseases and brain diseases is increasing, along with research on prevention, treatment, alleviation and improvement of such aging and diseases. It has been continued. However, the existing substances have problems such as unknown effects and induction of side effects, and it is necessary to develop a natural product-derived therapeutic agent that solves such problems.

U.S. Patent No. 9061023 U.S. Patent Publication No. 2002-0151506

LIM, H. J. et al., “Green Tea Catechin Leads to Global Improvement along Alzheimer's Disease-Related Phenotypes in NSE / hAPP-C105 Tg Mice”, Journal of Nutritional Biochemistry, 2013, vol. 24, no. 7, pages 1302-1313 CHOI, Y. T. et al., “The Green Tea Polyphenol (-)-epigallocatechin Gallate Attenuates beta-amyloid-induced Neurotoxicity in Cultured Hippocampal Neurons”, Life Sciences, 2001, vol. 70, pages 603-614 MPOFU, S. et al., “Interactive Efficacies of Elephantorrhiza Elephantina and Pentanisia Prunelloides Extracts and Isolated Compounds against Gastrointestinal Bacteria”, South African Journal of Botany, 2014, vol. 94, pages 224-230

  In one aspect, the present invention relates to a substance for improving cognitive function deterioration derived from a natural product, and an object thereof is to provide a composition for improving cognitive function deterioration including a post-fermented tea fraction.

  In another aspect, the present invention relates to 3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflavone (3-O-galloyl-3,3 'isolated from post-fermented tea fractions. It is an object of the present invention to provide a composition for improving cognitive function deterioration, which comprises (5, 5, 5 ', 7-pentahydroxyflavan) as an active ingredient.

  MEANS FOR SOLVING THE PROBLEMS In order to solve the above problems, the present invention provides, in one aspect, 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavone (3-O-garyl-3,3 ', 5 , 5 ', 7-pentahydroxyflavan), an isomer thereof, a pharmaceutically acceptable salt thereof, a prodrug thereof, a hydrate thereof, a solvate thereof, or a post-fermented tea ketone fraction containing it as an active ingredient Provided as a composition for improving cognitive dysfunction.

  The present invention also provides, in one aspect, 3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflavone (3-O-garyl-3,3 ', 5,5', 7- An effective amount of pentahydroxyflavan), an isomer thereof, a pharmaceutically acceptable salt thereof, a prodrug thereof, a hydrate thereof, a solvate thereof, or a post-fermented tea ketone fraction containing the same, which is required The present invention provides a method for improving and treating cognitive decline, which comprises administering to a subject.

  In another aspect, the present invention provides 3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflavone (3-O-garyl-3,3 ', 5,5', 7). -Pentahydroxyflavan), its isomer, its pharmaceutically acceptable salt, its prodrug, its hydrate, its solvate, or post-fermented tea ketone fraction containing it for improving cognitive function deterioration and treatment It provides an application for use in the manufacture of a composition.

  Furthermore, the present invention relates, in another aspect, to 3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflavone (3-O-gallyl-3, 3) for improvement and treatment of cognitive decline. 3 ', 5,5', 7-pentahydroxyflavan), its isomer, its pharmaceutically acceptable salt, its prodrug, its hydrate, its solvate, or post-fermented tea ketone fraction containing it Provide the goods.

  The present invention provides, in one aspect, a composition comprising a post-fermented tea ketone fraction and a composition comprising a specific compound, which can be widely utilized in cognitive function improvement related fields and neuroregressive disease management fields.

It is the result of analyzing a post-fermented tea extract by high performance liquid chromatography, and shows the case of passing an acetone fraction and the case of not passing an acetone fraction. The caffeine-removed chloroform: methanol 10: 1 (v / v) fraction was separated into 10 lower fractions using high-performance countercurrent chromatography (HPCCC) It shows that. It is shown for the fractions 1 to 5 of the 10 lower fractions fractionated using HPCCC. Of the 10 lower fractions fractionated using HPCCC, it is shown for fractions 6 to 10. It is a thing showing the inhibitory effect of Trans-3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavone on beta-amyloid aggregation. 1 shows a nuclear magnetic resonance (NMR) graph of Trans-3-O-galloyl-3,3 ′, 5,5 ′, 7-pentahydroxyflavone.

  Hereinafter, the present specification will be described in detail.

  As used herein, "post-fermented tea" includes those fermented by additional microorganisms or substances that are not enzymes present in the tea leaves.

  In the present specification, the "ketone fraction" is a fraction of a specific substance or an extract fractionated or fractionated using a ketone solvent, and a fraction thereof which is extracted again with a particular solvent. including. The type of the ketone is not limited, but acetone is preferred, and the fractionation method and extraction method may be any method known to those skilled in the art.

  As used herein, “isomers” are especially optical isomers (eg essentially pure enantiomers, essentially pure diastereomers or these Mixtures of) as well as conformational isomers (ie, isomers that differ only in their angle of one or more chemical bonds), positional isomers (especially tautomers) Or geometric isomers (eg, cis-trans isomers).

  In the present specification, the term "essentially pure" means, for example, a specific compound which can be mentioned as an enantiomer or a diastereomer when used in connection with an enantiomer or a diastereomer, for example. Is about 90% or more, preferably about 95% or more, more preferably about 97% or more, or about 98% or more, still more preferably about 99% or more, most preferably about 99.5% or more w / w) means to exist.

  As used herein, "pharmaceutically acceptable" is intended for use in animals, more particularly humans, by avoiding considerable toxicity when employed in conventional pharmaceutical doses. Can be approved by the government or equivalent regulatory body, or be recognized as being approved, or listed in the pharmacopoeia, or otherwise described in the general pharmacopoeia. Do.

  As used herein, "pharmaceutically acceptable salt" refers to a salt according to one aspect of the present invention which is pharmaceutically acceptable and which possesses the preferred pharmacological activity of the parent compound. The salt is formed of an inorganic acid such as (1) hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid or the like; or acetic acid, propionic acid, hexanoic acid, cyclopentene propionic acid, glycolic acid, pyruvic acid, Lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3- (4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo [2,2 , 2] -oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, tri Acid addition salt formed by an organic acid such as tyl acetate, tert-butyl acetate, lauryl sulfate, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid; or (2) parent compound It may include salts formed when the acidic protons present are replaced.

  As used herein, the term "prodrug" refers to a drug that has been chemically altered to physically or chemically modify a certain drug, and itself does not exhibit physiological activity, but administration It can later be turned into an original drug and exert its medicinal effects by the action of a chemical, physiological or enzyme in the body.

  In the present specification, "hydrate" means a compound to which water is bound, and is a broad concept including an encapsulated compound having no chemical bond between water and the compound. .

  As used herein, "solvate" refers to a higher-order compound generated between molecules or ions of a solute and molecules or ions of a solvent.

  The present invention relates, in one aspect, to 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavone (3-O-garyl-3,3 ', 5, represented by the following chemical formula 1 5 ', 7-pentahydroxyflavan), an isomer thereof, a pharmaceutically acceptable salt thereof, a prodrug thereof, a hydrate thereof, a solvate thereof, or a post-fermented tea ketone fraction containing it as an active ingredient Provided is a composition for improving cognitive dysfunction.

  According to one embodiment, the post-fermented tea ketone fraction may be a ketone fraction to a water-insoluble extract of post-fermented tea.

  According to another embodiment, the 3-O-galoyl-3,3 ′, 5,5 ′, 7-pentahydroxyflavone is a compound represented by Formula 2 below: Trans-3-O-galoyl-3,3 ', 5,5', 7-Pentahydroxyflavone (Trans-3-O-garyl-3,3 ', 5,5', 7-pentahydroxyflavan).

  According to another embodiment, the ketone fraction may be an alcohol extract of the ketone fraction.

  According to another embodiment, the ketone is acetone, carbon (carvon), pulegone, isolongifolnone, 2-heptanone, 2-pentanone, 3-hexanone, 3-heptanone, 4-heptanone, 2 -Octanone, 3-octanone, 2-nonanone, 3-nonanone, 2-undecanone, 2-tridecanone, methyl isopropyl ketone, ethyl isoamyl ketone, butylidene acetone, methyl heptenone, dimethyl octenone, geranyl acetone, farnesyl acetone, 2, 3 -Pentadione, 2,3-hexadione, 3,4-hexadione, 2,3-heptadione, amyl cyclopentanone, amyl cyclopentenone, 2-cyclopentyl cyclopentanone, hexyl cyclope Tanone, 2-n-heptylcyclopentanone, cis-jasmone, dihydrojasmone, methylcorrion, 2-tert-butylcyclohexanone, p-tert-butylcyclohexanone, 2-sec-butylcyclohexanone, celery ketone, krypton, p-tert-pentyl Cyclohexanone, methylcyclocitron, nerone, 4-cyclohexyl-4-methyl-2-pentanone, oxide ketone, emoxyflone, methylnaphthyl ketone, α-methylanisulacetone, anicylacetone, p-methoxyphenylacetone, benzylideneacetone, p -Methoxyacetophenone, p-methylacetophenone, propiophenone, acetophenone, α-dynascon (Dynascone), iritone (lritone), Nonionone, Pseudoionone (Pseudoionone), Methylionone, Methyliritone, 2,4-Di-tert-Butylcyclohexanone, Allylionone, 2-Acetyl-3,3-di-methyl norbornane, Verbenone, Fenchon, Cyclo It may contain pentadecanone, cyclohexadecenone and the like, and may contain any of ketones and their mixtures as solvents which can be generally used in the art, preferably acetone.

  According to another embodiment, the alcohol may be a single or a mixture of alcohols generally used in the art, preferably ethanol.

  In another aspect, the present invention provides 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavone (3-O-garyl-3,3 ', 5,5', 7-pentahydroxyflavan And its isomer, its pharmaceutically acceptable salt, its prodrug, its hydrate, or its solvate as an active ingredient.

  According to one embodiment, the 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavone is a compound represented by Formula 2: Trans-3-O-galloyl-3,3 ' , 5,5 ', 7-Pentahydroxyflavone (Trans-3-O-garyl-3,3', 5,5 ', 7-pentahydroxyflavan).

  According to another embodiment, 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavone or Trans-3-O-galloyl-3,5,5 in said composition The 5 ', 7-pentahydroxyflavone, an isomer thereof, a pharmaceutically acceptable salt thereof, a prodrug thereof, a hydrate thereof, or a solvate thereof is 0.000001 based on the total mass of the composition. Mass% or more, 0.000005 mass% or more, 0.00001 mass% or more, 0.00005 mass% or more, 0.0001 mass% or more, 0.0005 mass% or more, 0.001 mass% or more, 0.005 mass % Or more, 0.01% by mass or more, 0.05% by mass or more, 0.1% by mass or more, 0.3% by mass or more, 0.5% by mass or more, 1% by mass or more, 1.5% by mass or more, 1.8 mass% or more, 2 mass% or more, 2.2 mass Or more, 2.5 mass% or more, 2.8 mass% or more, 3 mass% or more, 3.2 mass% or more, 3.5 mass% or more, 3.8 mass% or more, 4 mass% or more, 4.2 3 mass% or more, 4.5 mass% or more, 4.8 mass% or more, 5 mass% or less, 4.8 mass% or less, 4.5 mass% or less, 4.2 mass% or less, 3. 8% by mass or less, 3.5% by mass or less, 3.2% by mass or less, 3% by mass or less, 2.8% by mass or less, 2.5% by mass or less, 2.2% by mass or less, 2% by mass or less, 1.8 mass% or less, 1.5 mass% or less, 1.2 mass% or less, 1 mass% or less, 0.5 mass% or less, 0.3 mass% or less, 0.1 mass% or less, 0.05 % By mass or less, 0.01% by mass or less, 0.005% by mass or less, 0.001% by mass or less, 0.0005% by mass or less, 0.0001% by mass or less, 0.00005 quality % Or less, 0.00001% by weight or less, or 0.000005 may be less by mass%. Preferably, it may be 0.000001 to 5% by mass or less.

  According to another embodiment, the content of the post-fermented tea ketone fraction in the composition is 0.01% by mass or more, 0.1% by mass or more, 1% by mass, based on the total mass of the composition. More than, 5 mass% or more, 10 mass% or more, 15 mass% or more, 20 mass% or more, 25 mass% or more, 30 mass% or more, 35 mass% or more, 40 mass% or more, 45 mass% or more, 50 mass% or more Or more, 55 mass% or more, 60 mass% or more, 65 mass% or more, 70 mass% or more, 75 mass% or more, 80 mass% or more, 85 mass% or more, 90 mass% or more, or 95 mass% or more 100% by mass or less, 95% by mass or less, 90% by mass or less, 85% by mass or less, 80% by mass or less, 75% by mass or less, 70% by mass or less, 65% by mass or less, 60% by mass or less, 55% by mass or less 50% by mass or less 45% by mass or less 40% by mass or less 5 mass% or less, 30 mass% or less, 25 mass% or less, 20 mass% or less, 15 mass% or less, 10 mass% or less, 5 mass% or less, 1 mass% or less, 0.1 mass% or less, or 0. It may be 05% by mass or less. Preferably, it may be 0.01% by mass to 100% by mass.

  According to another embodiment, said fraction is 3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflavone or Trans-3-O-galloyl-3,3'5 , 5 ', 7-pentahydroxyflavone, an isomer thereof, a pharmaceutically acceptable salt thereof, a prodrug thereof, a hydrate thereof, or a solvate thereof, is present in an amount of 0. 0 based on the total mass of the fractions. 0001 mass% or more, 0.0005 mass% or more, 0.001 mass% or more, 0.005 mass% or more, 0.01 mass% or more, 0.05 mass% or more, 0.1 mass% or more, 0.5 Mass% or more, 1 mass% or more, 1.5 mass% or more, 2 mass% or more, 2.5 mass% or more, 3 mass% or more, 3.5 mass% or more, 4 mass% or more, or 4.5 mass % Or more, 5% by mass or less, 4.5% by mass or less, 4% by mass or less, 3.5% by mass or less, Mass% or less, 2.5% by mass or less, 2% by mass or less, 1.5% by mass or less, 1% by mass or less, 0.5% by mass or less, 0.1% by mass or less, 0.05% by mass or less, 0 .01 mass% or less, 0.005 mass% or less, 0.001 mass% or less, or 0.0005 mass% or less. Preferably, 0.0001 mass% to 5 mass% may be contained.

  According to another embodiment, 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavone or Trans-3-O-galloyl-3,3'5 by administration of the composition The dose of 5,5,7-pentahydroxyflavone, an isomer thereof, a pharmaceutically acceptable salt thereof, a prodrug thereof, a hydrate thereof, or a solvate thereof is 0.0001 g / kg / day or more , 0.001 g / kg / day or more, 0.01 g / kg / day or more, 0.1 g / kg / day or more, 1 g / kg / day or more, 2 g / kg / day or more, 3 g / kg / day or more, 4 g / Kg / day or more, 5 g / kg / day or more, 6 g / kg / day or more, 7 g / kg / day or more, 8 g / kg / day or more, 9 g / kg / day or more, 10 g / kg / day or more, 11 g / day kg / day or more, 12 g / kg / day or more, 13 g / kg / day or more, or 14 g / kg / day or more, 15 g / kg / day or less, 14 g / kg / day or less, 13 g / kg / day or less, 12 g / kg / day or less, 11 g / kg / day or less, 10 g / kg / day 9 g / kg / day or less, 8 g / kg / day or less, 7 g / kg / day or less, 6 g / kg / day or less, 5 g / kg / day or less, 4 g / kg / day or less, 3 g / kg / day or less 2 g / kg / day or less, 1 g / kg / day or less, 0.1 g / kg / day or less, 0.01 g / kg / day or less, 0.001 g / kg / day or less, or 0.0005 g / kg / day It may be the following. Preferably, it may be 0.0001 g / kg / day to 15 mg / kg / day.

  According to another embodiment, the cognitive decline includes aggregation of beta-amyloid (beta-amyloid), formation of beta-amyloid plaque, brain-derived neurotrophic factor (BDNF). And at least one selected from the group consisting of decreased expression of DNMT1 (DNA (cytosine-5) -methyltransferase 1) and increased expression thereof.

  According to another embodiment, the cognitive decline may include one or more selected from the group consisting of memory decline, cognitive decline, cognitive decline, depression, and amnesia.

  According to another embodiment, the improvement is achieved by one or more selected from the group consisting of suppression of aggregation of beta-amyloid, degradation of aggregated beta-amyloid, enhancement of BDNF expression, and reduction of expression of DNMT1. You may

  A beta-Amyloid aggregation assay was performed using the Trans-3-O-galloyl-3,3 ′, 5,5 ′, 7-pentahydroxyflavone according to an embodiment of the present invention. As a result, the comparative substance epicatechin 3-O-gallate (Epicatechin 3-O-gallate, ECG, 1 μg / ml) has a 17% aggregation inhibitory effect compared to the control group (positive control) not treated with the compound. Trans-3-O-galoyl-3,3 ′, 5,5 ′, 7-pentahydroxyflavone exhibited a 64% aggregation inhibitory effect as compared to the control group. In particular, the beta-amyloid of Trans-3-O-galoyl-3,3 ′, 5,5 ′, 7-pentahydroxyflavone is a phenol known as an excellent aggregation inhibitor on the result graph (see FIG. 5). The better aggregation inhibitory effect was shown as it was located between red (phenol red, 55% aggregation inhibitory effect) and morin (morin, 78% aggregation inhibitory effect). Furthermore, in the Trans-3-O-galloyl-3,3 ′, 5,5 ′, 7-pentahydroxyflavone-treated group, the aggregation is reduced after a reaction time of 60 minutes, so Trans-3 It can be seen that -O-galoyl-3,3 ', 5,5', 7-pentahydroxyflavone is also effective in disassembling aggregated beta-amyloid. Therefore, it was confirmed that the trans-3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavone of the present invention can be used to improve the cognitive decline related to beta-amyloid. In addition, Trans-3-O-Galloyl-3,3 ′, 5,5 ′, 7-pentahydroxyflavone is used for the prevention and treatment of diseases caused by beta-amyloid aggregation (including neurodegenerative diseases such as Alzheimer's disease). It confirmed that it could be used effectively.

  In addition, Trans-3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflavone of the present invention enhanced BDNF expression and decreased DNMT1 expression in neuronal cells. That is, the present inventors have found out that the present invention can be effectively used for the prevention and treatment of cognitive decline related to BDNF expression reduction or DNMT1 expression increase, and neurodegenerative diseases such as Alzheimer's disease.

  In still another aspect of the present invention, the composition may be a pharmaceutical composition for preventing or treating neurodegenerative diseases.

  According to one embodiment, the neurodegenerative disease is selected from the group consisting of aggregation of beta-amyloid (β-amyloid), formation of plaques of beta-amyloid, decreased expression of BDNF, and increased expression of DNMT1. It may be caused by any one or more of the above.

  According to another embodiment, the neurodegenerative disease may include Alzheimer's disease and may be accompanied by symptoms including the cognitive decline.

  The pharmaceutical composition according to one aspect of the present invention may be administered orally, parenterally, rectally, topically, percutaneously, intravenously, intramuscularly, intraperitoneally, subcutaneously and the like. The dosage form for oral administration may be, but is not limited to, tablets, pills, soft and hard capsules, granules, powders, fine granules, solutions, emulsions or pellets . Dosage forms for parenteral administration may be, but are not limited to, solutions, suspensions, emulsions, gels, drops, suppositories, patches or sprays. The dosage form may be easily manufactured by a method usual in the field, and may be a surfactant, an excipient, a wettable, an emulsion promoter, a suspension, a salt or buffer for regulating the osmotic pressure. Colorants, spices, stabilizers, preservatives, preservatives or other customary auxiliaries may further be included.

  The applied amount or dose of the pharmaceutical composition according to one aspect of the present invention may vary depending on the age, sex, weight, pathological condition and severity of the subject to be administered, the route of administration or the judgment of the prescriber. The determination of the applied dose based on such factors is within the level of those skilled in the art, and the daily dose of the active ingredient is, for example, 0.001 μg / kg / day to 5000 mg / kg / day, more specifically , 0.0001 mg / kg / day to 15 mg / kg / day, but is not limited thereto.

  According to another embodiment, the composition may be a food composition.

  Although the dosage form of the food composition is not particularly limited, for example, it is formulated into tablets, granules, pills, powders, solutions such as drinks, caramel, gel, bars, tea bags, etc. Good. The food composition of each dosage form may be properly selected and formulated by the person skilled in the art according to the dosage form or purpose of use, in addition to the active ingredient, according to the dosage form or purpose of use. When applied, synergy can be achieved.

  The composition may be administered in various ways, such as simple ingestion, drinking, injection, spray or squeeze administration.

  In the food composition according to one aspect of the present invention, the determination of the dose of the active ingredient is within the level of those skilled in the art, and the daily dose thereof is, for example, 0.0001 mg / kg / day to 15 mg / kg / day The day may be, but is not limited to, it may vary depending on various factors such as the age, health condition, and complications of the subject it is desired to administer.

  The food composition according to one aspect of the present invention is, for example, various foods such as chewing gum, caramel products, candies, frozen desserts, confectionery and the like, beverage products such as soft drinks, mineral water, alcoholic beverages, vitamins and minerals And health functional food products.

  In addition to the above, the food composition according to one aspect of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring agents and enhancers (cheese, chocolate, etc. Pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used for carbonated beverages, etc. You may In addition, the food composition according to one aspect of the present invention may include natural fruit juices and fruit juice beverages, and pulp for the production of vegetable beverages. These components may be used independently or in combination. The proportion of these additives is not particularly important, but is generally comprised in the range of 0 to about 50 parts by weight per 100 parts by weight of the composition according to one aspect of the present invention.

  Hereinafter, the configuration and effects of the present specification will be more specifically described by way of examples, experimental examples, and dosage form examples. These examples are merely illustrated for the purpose of assisting understanding of the present specification, and the scope and range of the present specification are not limited by the following examples.

<Example 1> Preparation of post-fermented tea sample Water was added to green tea made of green tea (Camellia sinensis var. Yabukita) leaves to adjust the water content to 40% by mass. This was inoculated with 5 × 10 ⁶ cfu / g of Bacillus subtilis and fermented at 50 ° C. for 3 days and then fermented at 80 ° C. for 4 days. The fermented tea was dried to a moisture content of 4% by mass to 6% by mass with hot air at 70 ° C to 80 ° C, and then aged at 10 ° C for 100 days.

  The aged tea sample was ground for 15 seconds and passed through a stainless steel sieve of 1 mm mesh size. Next, 50 mg of crushed is put into a 1.5 ml Eppendorf tube, 1 ml of deionized water is added and stirred at constant temperature for 30 minutes in a 60 ° C. constant temperature water bath, then at 13,000 rpm at 25 ° C. Centrifuge for 15 minutes. Only the portion insoluble in water was separated from the dried fermented green tea extract.

<Example 2> Acquisition of fraction and separation of compound The above-mentioned post-fermented tea sample of 150 g was fractionated with acetone to remove as much as possible the catechin derivative and caffeine, and the soluble matter enriched in flavonoid was obtained. A mixture of chloroform: methanol was used as a solvent to elute sequentially 10: 1, 5: 1, 1: 1 (v / v) to 40 g of the acetone solubles, using a silica gel column chromatography as a primary Each fraction was obtained.

  Here, as a result of analyzing 12 g of chloroform: methanol 10: 1 (v / v) fraction by high performance liquid chromatography (HPLC, Waters Alliance 2695 system, Waters, USA), acetone is used as described above. If not fractionated, caffeine was still present as the main compound, and it was confirmed that caffeine was removed after fractionation using acetone as described above (FIG. 1).

  Using high-performance countercurrent chromatography (HPCCC, Dynamic Extractions Ltd, UK), high-performance countercurrent chromatography (HPCCC) with 8.9 g of chloroform: methanol 10: 1 (v / v) fractions from which caffeine has been removed I fractionated. The solvent used at this time is n-hexane-TBME (Methyl tert-butyl ether) -BuOH-MeCN-Water (0.25: 3: 1: 1: 5, v / v), and the flow rate is 25 ml / min. Met. Using the above conditions, a total of 10 lower fractions were separated (Figure 2) and analyzed by HPLC to confirm that the majority of the compounds contained in each fraction were flavonoid-based ( Figure 3, Figure 4).

From the above fraction, Trans-3-O-galloyl-3,3 ′, 5,5 ′, 7-pentahydroxyflavone (Trans-3-O-gallyl-3,3 ′, 5,5 ′, 7-pentahydroxyflavan , C ₂₂ H ₁₈ O ₁₀ , molecular weight 442.0900), and the structure of the separated compound is as follows.

^{In the case of 1} H nuclear magnetic resonance (NMR), methanol-d3 is used as a solvent (solvent), the instrument is Bruker Advance DPX-500 (BRUKER, USA), Trans-3-O-Garoyl The NMR data of −3,3 ′, 5,5 ′, 7-pentahydroxyflavone are as follows, and a specific graph is as shown in FIG.

¹ H NMR d (500 Hz) 7.01 (s, 2 H), 6.89 (s, 1 H), 6.76 (s, 2 H), 6.04 (s, 1 H), 5.96 (s, 1 H) ), 5.36 (m, 1 H), 5.13 (m, 1 H), 2.83 (m, 1 H), 2.73 (m, 1 H)

<Experimental example 1> Confirmation of the beta-amyloid aggregation effect The beta-amyloid aggregation inhibitory effect of said Trans-3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavone is fluorometrically determined (Thioflavin T assay) Confirmed.

  Specifically, it was used at a concentration of 5 μM of beta-amyloid (Aβ 1-42, American Peptide Co, USA) and stored at -80 ° C before use. Morin (Morin, 100 μM), phenol red (Phenol Red, 100 μM), epicatechin 3-O-gallate (Epicatechin 3-O-gallate, ECG, 1 μg / ml) in DMSO, Trans-3-O-galoyl-3, Each of 3 ', 5,5', 7-pentahydroxyflavone (1 μg / ml) was diluted to adjust to the above concentration. In order to specify the degree of Aβ 1-42 aggregation inhibition, 5 μl of each compound prepared at the above concentration is added to 92.5 μl of 0.01 M sodium phosphate buffer solution, and 2.5 μl of 5 μM Aβ 1-42 is added, After reaction at 37 ° C. for 26 hours, 2,000 μl of 5 μM Thioflavin T (Thioflavin T) was added, and measurement was performed using a fluorescence spectrophotometer (RF-5300 PC, SHIMADZU CORPORATION, Japan).

  The results are also shown in FIG. That is, ECG and Trans-3-O-galoyl-3,3 ′, 5,5 ′, 7-pentahydroxyflavone each aggregate only beta amyloid without treating the compound as a positive control. Showed a 17% and 64% aggregation inhibitory effect compared to the In particular, the inhibitory effect (64%) of Trans-3-O-galoyl-3,3 ′, 5,5 ′, 7-pentahydroxyflavone is phenol red (55%) used as a positive inhibitor. The better aggregation inhibitory effect was shown as it was located between () and Morin (78%). In the Trans-3-O-galoyl-3,3 ′, 5,5 ′, 7-pentahydroxyflavone-treated group, from the appearance that aggregation is reduced after 60 minutes of reaction time, Trans-3-O-galoyl-3,3 It was confirmed that ', 5,5', 7-pentahydroxyflavone is also effective in disassembling aggregated beta-amyloid.

<Experimental Example 2> Confirmation of the Expression Amount of BDNF (Brain-Derived Neurotrophic Factor) and DNMT 1 (DNA (Cytosine-5) -Methyltransferase 1) in Cells SH-SY5Y (Neuroblastoma, Korea) Cell line bank) cell lines are seeded at 2 × 10 ⁶ per well in 6-well plates (well plate, FALCON), cultured for 24 hours in a 37 ° C., 5% CO ₂ incubator, GCG 10 μM, EGCG 10 μM, 10 μg / ml of existing green tea extract (GTE), 10 μg / ml of Trans-3-O-galoyl-3,3 ′, 5,5 ′, 7-pentahydroxyflavone, 5 as a positive control group -Aza- 'Deoxycytidine (5-Aza, Sigma-aldrich) was treated with 1μM 24 hours further incubation. Then, the medium was completely removed, and RNA was extracted using an RNA extraction kit (RNeasy mini kit, Quiagen). After quantifying the extracted mRNA using a UV detector (TECAN), 1 μg of mRNA was synthesized to a complementary DNA using a kit (SuperScript VILO cDNA Synthesis Kit, Thermofisher scientific). About 1 μg of complementary DNA was taken, and a real-time quantitative chain polymerization reaction was carried out using Taqman probe (Life technology) and Quantitect Probe PCR Kit (Quiagen). Thereby, the expression levels of BDNF and DNMT1 were confirmed. At this time, GAPDH which is a housekeeping gene was used as the correction reference mRNA.

  The expression levels of BDNF and DNMT1 are shown in Table 1 and Table 2, respectively.

<Dose formulation example 1> Soft capsule 150 mg of post-fermented tea acetone fraction according to Example 2 of the present invention, 440 mg of lactose, 430 mg of corn starch and 2 mg of magnesium stearate were mixed to prepare a soft capsule filling liquid. Then, separately from the filling solution, a soft capsule sheet was produced at a ratio of 66 parts by mass of gelatin, 24 parts by mass of glycerin and 10 parts by mass of sorbitol solution, and the filling solution was filled to produce a soft capsule.

  In addition, according to the example of the present invention, 3 mg of Trans-3-O-galoyl-3,3 ′, 5,5 ′, 7-pentahydroxyflavone, 440 mg of lactose, 450 mg of corn starch and 2 mg of magnesium stearate are mixed and soft. Produce a capsule filling solution. Then, a soft capsule sheet was manufactured as described above, and the filling liquid was filled to manufacture a soft capsule.

<Formulation Example 2> Tablet A post-fermented tea acetone fraction 150 mg, vitamin E 15 mg, vitamin C 15 mg, galactooligosaccharide 250 mg, lactose 60 mg and maltose 140 mg according to Example 2 of the present invention are mixed and fluidized bed dryer After granulation using, 8 mg of sugar ester was added. The composition was compressed into tablets by a conventional method.

  In addition, Trans-3-O-galoyl-3,3 ′, 5,5 ′, 7-pentahydroxyflavone 3 mg, vitamin E 15 mg, vitamin C 15 mg, galactooligosaccharide 259 mg, lactose 60 mg and maltose 140 mg were mixed according to the example. After granulation using a fluid bed dryer, 8 mg of sugar ester was added. The composition was compressed into tablets by a conventional method.

<Dose form example 3> Drink agent 80 mg of post-fermented tea acetone fractions according to Example 2 of the present invention, 9 mg of vitamin E, 9 mg of vitamin C, 10 g of glucose, 0.6 g of citric acid and 25 g of liquid oligosaccharide To this, 400 ml of purified water was added and the mixture was filled in a bottle. After filling the bottle, it was sterilized at 30 ° C. for 4 to 5 seconds to produce a drink.

  In addition, Trans-3-O-galoyl-3,3 ′, 5,5 ′, 7-pentahydroxyflavone 0.1 mg, vitamin E 9 mg, vitamin C 9 mg, glucose 10 g, citric acid 0.6 g, and the examples. Twenty-five grams of liquid oligosaccharides were mixed, 400 ml of purified water was added thereto, and each bottle was filled with 200 ml. After filling the bottle, it was sterilized at 30 ° C. for 4 to 5 seconds to produce a drink.

<Dose form example 4> Granules 150 mg of post-fermented tea acetone fraction according to Example 2 of the present invention, 9 mg of vitamin E, 9 mg of vitamin C, 250 mg of anhydrous crystalline glucose and 550 mg of starch are mixed and a fluid bed granulator is used The mixture was granulated into granules and filled into packets to produce granules.

  Moreover, Trans-3-O-galoyl-3,3 ′, 5,5 ′, 7-pentahydroxyflavone 3 mg, vitamin E 9 mg, vitamin C 9 mg, anhydrous crystalline glucose 250 mg and starch 550 mg according to the example are mixed and flowed After granulation into granules using a layer granulator, the package was filled to produce granules.

<Formulation Example 5> Injection According to the composition shown in Table 3 below, an injection was manufactured by a usual method.

  Also, instead of the post-fermented tea acetone fraction, 3 mg of Trans-3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflavone of Example was used to prepare an injection.

<Dose Formulation Example 6> Health Food Health food was manufactured according to the conventional method according to the composition shown in Table 4 below.

  Also, instead of the post-fermented tea acetone fraction, a health food was manufactured using 3 mg of Trans-3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflavone of Example.

  Although the composition ratio of the vitamin and mineral mixture is a mixture composition taking as an example a component that is relatively suitable for health food, the composition ratio may be arbitrarily modified and implemented, as in the method for producing a general health food. After mixing the ingredients described above, they may be used for producing health food compositions in a conventional manner.

  <Formulation Example 7> Healthy Beverage

  As shown in Table 5 above, the remaining amount of purified water was added to a total volume of 900 ml, and the above components were mixed according to a conventional method for producing a health drink, and then stirred and heated at 85 ° C for about 1 hour. After that, the prepared solution was filtered, put into a sterilized 2 liter container, sealed and sterilized, and then stored under refrigeration to produce a health drink.

  Also, instead of the post-fermented tea acetone fraction, 0.1 mg of Trans-3-O-galoyl-3,3 ′, 5,5 ′, 7-pentahydroxyflavone of Example is used to produce a health drink. did.

  While specific parts of the specification have been described above in detail, such specific descriptions are merely preferred embodiments for those of ordinary skill in the art, and this is the scope of the present description. It is obvious that is not limited. Accordingly, the substantial scope of the present specification can be said to be defined by the claims and their equivalents.

Claims (15)

3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflavone (3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavan) represented by the following chemical formula 1, A composition for improving cognitive function decline, comprising the isomer, a pharmaceutically acceptable salt thereof, a prodrug thereof, a hydrate thereof, a solvate thereof, or a post-fermented tea ketone fraction containing it as an active ingredient object.

  The composition according to claim 1, wherein the post-fermented tea ketone fraction is a ketone fraction with respect to a water-insoluble extract of post-fermented tea.   The composition according to claim 1, wherein the ketone is acetone. The 3-O-galoyl-3,3 ′, 5,5 ′, 7-pentahydroxyflavone is a compound represented by Formula 2 below: Trans-3-O-galloyl-3,3 ′, 5,5 ′, 7 The composition according to claim 1, which is -pentahydroxyflavone (Trans-3-O-garyl-3,3 ', 5,5', 7-pentahydroxyflavan).

  3-O-galoyl-3,3 ′, 5,5 ′, 7-pentahydroxyflavone in the composition, an isomer thereof, a pharmaceutically acceptable salt thereof, a prodrug thereof, a hydrate thereof or The composition according to claim 1, wherein the content of the solvate is 0.000001% by mass to 5% by mass with respect to the total mass of the composition.   The composition according to claim 1, wherein the content of the post-fermented tea ketone fraction in the composition is 0.01% by mass to 100% by mass with respect to the total mass of the composition.   Said fraction is said 3-O-galloyl-3,3 ', 5,5', 7-pentahydroxyflavone, its isomer, its pharmaceutically acceptable salt, its prodrug, its hydrate The composition according to claim 1, wherein or the solvate is contained in an amount of 0.0001% to 5% by mass based on the total mass of the fraction.   3-O-galoyl-3,3 ', 5,5', 7-pentahydroxyflavone (3-O-garyl-3,5,5 ', 7-pentahydroxyflavan) by administration of the composition, The dose of an isomer, a pharmaceutically acceptable salt thereof, a prodrug thereof, a hydrate thereof, or a solvate thereof is 0.0001 mg / kg / day to 15 mg / kg / day. The composition as described in.   The cognitive decline includes aggregation of beta-amyloid, decreased expression of brain-derived neurotrophic factor (BDNF), and expression of DNMT1 (DNA-5 (cytosine-5) -methyltransferase 1). The composition according to any one of claims 1 to 8, which is due to one or more selected from the group consisting of an increase.   The composition according to any one of claims 1 to 8, wherein the cognitive decline includes one or more selected from the group consisting of memory decline, cognitive decline, cognitive decline, depression, and amnesia.   The improvement is achieved by one or more selected from the group consisting of suppression of aggregation of beta-amyloid, degradation of aggregated beta-amyloid, enhancement of BDNF expression, and reduction of DNMT1 expression. Composition as described.   The composition according to any one of claims 1 to 8, wherein the composition is a pharmaceutical composition for preventing or treating neurodegenerative diseases.   The pharmaceutical composition according to claim 1, wherein the neurodegenerative disease is caused by one or more selected from the group consisting of aggregation of beta-amyloid (β-amyloid), decreased BDNF expression, and increased DNMT1 expression. 12. The composition according to 12.   The composition according to claim 12, which is a pharmaceutical composition, wherein the neurodegenerative disease is Alzheimer's disease.   The composition according to any one of claims 1 to 8, wherein the composition is a food composition.

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