CN108697687A

CN108697687A - Including 3-O- galloyls -3,3 ', the cognitive function improvement composition of 5,5 ', 7- penta hydroxy group flavane

Info

Publication number: CN108697687A
Application number: CN201780013673.2A
Authority: CN
Inventors: 洪勇德; 崔敏植; 曹始永; 金正基
Original assignee: Amorepacific Corp
Current assignee: Amorepacific Corp
Priority date: 2016-07-18
Filing date: 2017-07-17
Publication date: 2018-10-23
Also published as: JP2019521076A; JP7116682B2; KR102441381B1; KR20180009316A

Abstract

One aspect of the present invention is related to the low improvement composition of cognitive function as active ingredient comprising post-fermented tea ketone fraction, is on the other hand related to including the 3-O- galloyls -3,3' detached from post-fermented tea ketone fraction;,5,5';, 7- penta hydroxy group flavane (3-O-galloyl-3,3';,5,5';7-pentahydroxyflavan), the low improvement composition of cognitive function of its isomers, its pharmaceutically acceptable salt, its prodrug, its hydrate or its solvate as active ingredient, utilize an aspect of of the present present invention, the aggregation of beta-amyloid protein can be effectively inhibited, BDNF expression and DNMT1 expression can be adjusted, therefore the present invention can be applied to and the low relevant various fields of cognitive function on the one hand.

Description

Including 3-O- galloyls -3,3 ', the cognitive function of 5,5 ', 7- penta hydroxy group flavane, which improves, to be used Composition

Technical field

This specification is related to a kind of including 3-O- galloyls -3,3',5,5', 7- penta hydroxy group flavane (3-O-galloyl-3,3', 5,5', 7-pentahydroxyflavan) cognitive function improvement composition.

Background technology

With the improvement of living standards with the development of medical industries, using cancer as the treatment of the curative disease of hardly possible of representative at To be possible, increase to the service life of the mankind, but thus the aging of the caused mankind has caused chronic degenerative nervous system disease The increase of disease, this relative reduction quality of life instead.Nerve cell dysfunction and damage may be by the albumen of easy cohesion Matter and induce, most of the nervous system disease is characterized by this symptom.This nervous system disease includes such as alzheimer ' The diseases such as silent disease.

Alzheimer disease refer to the neural cell injury of brain and show cognitive ability decline, failure of memory, The diseases such as the irreversible loss of memory, disorientation, recognition capability decline, depression, amnesia and language competence impairment disorders The progressive neurodegenerative disease of shape.Alzheimer disease be after cranial vascular disease, cancer it is the third-largest with it is aging The relevant cause of death, mean disease duration is 10 years or more when morbidity, therefore the spirit of guardian, financial burden and society Increase can be born.The most representative reason of known Alzheimer disease can enumerate beta-amyloid protein (β-so far Amyloid, A β, also known as amyloid beta) improper aggregation (aggregation).That is, big intracerebral normal presence Beta-amyloid protein is abnormally assembled and forms amyloid plaques (A β plaque), and thus nerve cell is connected with nerve cell It damages and falls ill.

As elderly population increase, for the needs of the treatment and prevention of aging, degenerative neurological diseases and cerebral disorders Increase, thus the research for preventing, treating, alleviating and improving etc. of this aging or disease is persistently carried out, but existing object Matter effect is indefinite or there is the problems such as inducing side effect, it is therefore desirable to which exploitation solves the problems, such as this controlling from natural materials It is truth to treat agent.

Invention content

The present invention is related to the lowly improvement substance of the cognitive function from natural materials on the one hand, as a result, to provide packet The cognitive function of the fraction containing post-fermented tea is lowly for the purpose of improvement composition.

The present invention is on the other hand to provide comprising the 3-O- galloyls -3,3&apos detached from post-fermented tea fraction;,5,5',7- Penta hydroxy group flavane (3-O-galloyl-3,3',5,5', 7-pentahydroxy flavan) and cognitive function as active ingredient For the purpose of low improvement composition.

In order to solve the above problems, the present invention provides cognitive function lowly improvement composition on the one hand, contains 3- O- galloyls -3,3',5,5', 7- penta hydroxy group flavane (3-O-galloyl-3,3',5,5', 7-pentahydroxyflavan), its Isomers, its pharmaceutically acceptable salt, its prodrug, its hydrate, its solvate or include its post-fermented tea ketone point Object is evaporated as active ingredient.

In addition, the present invention provides on the one hand improves and treats the low method of cognitive function comprising by 3-O- galloyls- 3,3',5,5', 7- penta hydroxy group flavane (3-O-galloyl-3,3',5,5', 7-pentahydroxyflavan), its isomers, Its pharmaceutically acceptable salt, its prodrug, its hydrate, its solvate or include its post-fermented tea ketone fraction Effective quantity is administered to needing its individual.

In addition, the present invention provides 3-O- galloyls -3,3&apos on the other hand;,5,5', 7- penta hydroxy group flavane (3-O-galloyl- 3,3',5,5', 7-pentahydroxyflavan), its isomers, its pharmaceutically acceptable salt, its prodrug, its hydrate, Its solvate or lowly improve and the manufacture of therapeutic composition in cognitive function comprising its post-fermented tea ketone fraction In purposes.

Moreover, the present invention provides the 3-O- galloyl -3,3&apos low for improving and treating cognitive function on the other hand;,5, 5', 7- penta hydroxy group flavane (3-O-galloyl-3,3',5,5', 7-pentahydroxyflavan), its isomers, its pharmaceutically Acceptable salt, its prodrug, its hydrate, its solvate or the post-fermented tea ketone fraction comprising it.

The present invention provides the composition comprising post-fermented tea ketone fraction and the combination comprising specific compound on the one hand Object, so as to be widely used in and improve the relevant field of cognitive function and neurodegenerative disease management domain.

Description of the drawings

Fig. 1 be with high performance liquid chromatography to post-fermented tea extract analyzed as a result, indicating to be fractionated by acetone Situation and the case where be fractionated without acetone.

Fig. 2 indicates to utilize high capacity highly effective adverse current chromatogram (high-performance countercurrent Chromatography, HPCCC) chloroform of caffeine will be eliminated:Methanol 10:1 (v/v) fraction is divided into 10 sub- fractions.

Fig. 3 indicates the fraction 1 to 5 in the above-mentioned 10 sub- fractions using HPCCC fractionation.

Fig. 4 indicates the fraction 6 to 10 in the above-mentioned 10 sub- fractions using HPCCC fractionation.

Fig. 5 indicates the trans- -3-O- galloyls -3,3&apos for beta-amyloid aggregation;,5,5', the suppression of 7- penta hydroxy group flavane Effect processed.

Fig. 6 indicates trans- -3-O- galloyls -3,3',5,5', nuclear magnetic resonance (NMR) chart of 7- penta hydroxy group flavane.

Specific implementation mode

In the following, being described in detail to this specification.

In this specification " post-fermented tea " include using the other microorganisms or substance for being not present in enzyme in tealeaves into The tea of row fermentation.

In this specification, " ketone fraction " includes object obtained by being fractionated to predetermined substance or extract using ketone solvent Remaining substance after matter or fractionation, also comprising substance obtained by extracting these again with specific solvent.The type of above-mentioned ketone can To be any, but preferably acetone, any means that fractional method and extracting method can be known to the skilled person.

In this specification, " isomers " includes not only especially optical isomer (optical isomers) (for example, base Pure mirror image isomer (essentially pure enantiomers), substantially pure partial stereoisomer in sheet (essentially pure diastereomers) or their mixture), also include rotamer (conformation Isomers) (that is, isomers that more than one chemical bond only has its angle different), position isomer (position Isomers) (especially tautomer (tautomers)) or geometric isomer (geometric isomers) are (for example, suitable Formula-transisomer).

In this specification, " substantially pure (essentially pure) " refers to for example about mirror image isomer or part In the case that isomers carries out use, the particular compound that can be used as mirror image isomer or partial allosteric body and enumerate exists about 90% or more, preferably from about 95% or more, even more preferably about 97% or more or about 98% or more, more preferably about 99% with On, still more preferably it is about 99.5% or more (w/w).

In this specification, " pharmaceutically acceptable " refers to common medicine and pharmacology pharmaceutical dosage (medic inal When dosage) utilizing, by avoiding comparable toxic effect, can be used in animal, more specifically can be used in the mankind can It obtains the approval of government or the regulatory agency of its approval or has got the Green Light, or be acknowledged as enumerating or being recorded in pharmacopeia Situation in other general pharmacopeia.

" pharmaceutically acceptable salt " refers to pharmaceutically acceptable and has parent compound (parent in this specification Compound the salt according to an aspect of the present invention of preferred pharmacological activity).Above-mentioned salt can include (1) by such as hydrochloric acid, hydrogen The inorganic acids such as bromic acid, sulfuric acid, nitric acid, phosphoric acid are formed;Or by such as acetic acid, propionic acid, caproic acid, pentamethylene propionic acid, glycolic, acetone Acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3- (4- hydroxy benzenes first Acyl group) benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2- ethionic acids, 2- isethionic acids, benzene sulfonic acid, 4- chlorine Benzene sulfonic acid, 2- naphthalene sulfonic acids, 4- toluenesulfonic acids, camphorsulfonic acid, 4- Jia Jishuanhuans [2,2,2]Oct-2-ene -1- carboxylic acids, glucoheptose Acid, 3- phenylpropionic acids, trimethylace tonitric, butylacetic acid, lauryl sulfate, gluconic acid, glutamic acid, hydroxynaphthoic acid, water The acid-addition salts (acid addition salt) that the organic acids such as poplar acid, stearic acid, muconic acid are formed;Or (2) are present in mother The salt that acid proton in body compound is formed when substituted.

In this specification, " prodrug (prodrug) " is to instigate certain drug that chemical change occurs and adjust physical property, change The drug for learning property can pass through chemistry, physiology or enzyme in vivo although itself will not show physiological activity after administration Effect and become original drug to play drug effect.

In this specification, " hydrate (hydrate) refers to the compound for being combined with water, is comprising between water and compound There is no the extensive concept of the interior packet compound of chemical binding force.

In this specification, " solvate " refer to solute molecule or ion and the molecule or ion of solvent between generate High-orderCompound.

An aspect of of the present present invention provides a kind of cognitive function lowly improvement composition, and it includes by 1 table of following chemical formula 3-O- galloyls-the 3,3&apos shown;,5,5', 7- penta hydroxy group flavane (3-O-galloyl-3,3',5,5',7- Pentahydroxyflavan), its isomers, its pharmaceutically acceptable salt, its prodrug, its hydrate, its solvate or Including its post-fermented tea ketone fraction is as active ingredient.

[Chemical formula 1]

According to a concrete example, above-mentioned post-fermented tea ketone fraction can be the water-insoluble extract for post-fermented tea Ketone fraction.

According to another concrete example, above-mentioned 3-O- galloyls -3,3',5,5', 7- penta hydroxy groups flavane can be by following chemical formula 2 Trans- -3-O- galloyls-the 3,3&apos indicated;,5,5', 7- penta hydroxy group flavane (Trans-3-O-galloyl-3,3',5,5',7- pentahydroxyflavan)。

[Chemical formula 2]

According to other concrete examples, above-mentioned ketone fraction can be the alcohol extracting thing of ketone fraction.

According to other concrete examples, above-mentioned ketone can include acetone, carvol (carvon), pulegone (pulegone), Isolongitolanone (isolongifolanone), 2-HEPTANONE, 2 pentanone, 3- cyclohexanone, 3- heptanone, 4- heptanone, methyln-hexyl ketone, 3- are pungent Ketone, methyl n-heptyl ketone, 3- nonanones, methyln nonyl ketone, methyl n-undecyl ketone, methyl isopropyl Ketone, ethyl isoamyl ketone, butylideneacetone, Methyl heptenone, dimethyl-octa ketenes, geranyl acetone, farnesyl acetone, 2,3- pentanediones, 2,3- cyclohexanediones, 3,4- Acetyl butyryl, 2,3- heptadione, amyl cyclopentanone, amyl cyclopentenone, 2- cyclopenta rings pentanone, hexyl cyclopentanone, 2- n-heptyls Cyclopentanone, cis-jasmone, dihydro jasmone, methyl cyclopentenyl ketone2- tert-butyl cyclohexanones, Para-tertiary butyl cyclohexanone, 2- sec-butyl cyclohexanones, apione, cryptone, orivone, 1- (3,5,6- trimethyls -3- Cyclohexene -1- bases) ethyl ketone, nerone, 4- cyclohexyl -4-methyl-2 pentanone, oxydone, epoxy IsophoroneMethyl naphthyl ketone, Alpha-Methyl anisal acetone, Anisylacetone, to methoxybenzene Benzylacetone, BENZYLIDENE ACETONE, acetanisole, melilotal, propiophenone, acetophenone, α-imperial court's ketone (Dynascone), iritone (lritone), ionone (ionone), pseudoionone (Pseudoionone), irone, Methyl iritone, 2,4- di-t-butyls cyclohexanone, allyl ionone, 2- acetyl group -3,3- dimethyl norbornane, Verbena officinalis Ketenes, fenchone (fenchon), cyclopentadecanone, ring hexadecene ketone etc. can include that all can usually make as in the field The mixture of the ketone of solvent and they can be preferably acetone.

According to another concrete example, above-mentioned alcohol can be the field usually workable individually alcohol or mixture, can be preferred For ethyl alcohol.

It includes 3-O- galloyls -3,3&apos that the present invention provides on the other hand;,5,5', 7- penta hydroxy group flavane (3-O-galloyl- 3,3',5,5', 7-pentahydroxyflavan), its isomers, its pharmaceutically acceptable salt, its prodrug, its hydrate, Or cognitive function low improvement composition of its solvate as active ingredient.

According to a concrete example, above-mentioned 3-O- galloyls -3,3',5,5', 7- penta hydroxy groups flavane can be the anti-of above-mentioned chemical formula 2 Formula -3-O- galloyls -3,3',5,5', 7- penta hydroxy group flavane (Trans-3-O-galloyl-3,3',5,5',7-pentahydro xyflavan)。

According to another concrete example, 3-O- galloyls -3,3&apos in above-mentioned composition;,5,5', 7- penta hydroxy groups flavane or trans- -3- O- galloyls -3,3',5,5', 7- penta hydroxy groups flavane, its isomers, its pharmaceutically acceptable salt, its prodrug, its hydrate or Its solvate on the basis of above-mentioned composition total weight can be 0.000001 weight % or more, 0.000005 weight % with Upper, 0.00001 weight % or more, 0.00005 weight % or more, 0.0001 weight % or more, 0.0005 weight % or more, 0.001 weight % or more, 0.005 weight % or more, 0.01 weight % or more, 0.05 weight % or more, 0.1 weight % or more, 0.3 weight % or more, 0.5 weight % or more, 1 weight % or more, 1.5 weight % or more, 1.8 weight % or more, 2 weight % with Upper, 2.2 weight % or more, 2.5 weight % or more, 2.8 weight % or more, 3 weight % or more, 3.2 weight % or more, 3.5 weights Measure % or more, 3.8 weight % or more, 4 weight % or more, 4.2 weight % or more, 4.5 weight % or more or 4.8 weight % with On or 5 weight % or less, 4.8 weight % or less, 4.5 weight % or less, 4.2 weight % or less, 3.8 weight % or less, 3.5 weight % or less, 3.2 weight % or less, 3 weight % or less, 2.8 weight % or less, 2.5 weight % or less, 2.2 weight % Below, 2 weight % or less, 1.8 weight % or less, 1.5 weight % or less, 1.2 weight % or less, 1 weight % or less, 0.5 weight Measure % or less, 0.3 weight % or less, 0.1 weight % or less, 0.05 weight % or less, 0.01 weight % or less, 0.005 weight Measure % or less, 0.001 weight % or less, 0.0005 weight % or less, 0.0001 weight % or less, 0.00005 weight % or less, 0.00001 weight % or less or 0.000005 weight % or less.Preferably 0.000001 to 5 weight %.

According to other concrete examples, in above-mentioned composition, the content of post-fermented tea ketone fraction is relative to above-mentioned composition Total weight can be 0.01 weight % or more, 0.1 weight % or more, 1 weight % or more, 5 weight % or more, 10 weight % with Upper, 15 weight % or more, 20 weight % or more, 25 weight % or more, 30 weight % or more, 35 weight % or more, 40 weight % Above, 45 weight % or more, 50 weight % or more, 55 weight % or more, 60 weight % or more, 65 weight % or more, 70 weights % or more, 75 weight % or more, 80 weight % or more, 85 weight % or more, 90 weight % or more or 95 weight % or more are measured, Or 100 below weight %, 95 weight % or less, 90 weight % or less, 85 weight % or less, 80 weight % or less, 75 weights Measure % or less, 70 weight % or less, 65 weight % or less, 60 weight % or less, 55 weight % or less, 50 weight % or less, 45 Weight % or less, 40 weight % or less, 35 weight % or less, 30 weight % or less, 25 weight % or less, 20 weight % or less, 15 weight % or less, 10 weight % or less, 5 weight % or less, 1 weight % or less, 0.1 weight % or less or 0.05 weight % Below.Can be preferably 0.01 weight % to 100 weight %.

According to other concrete examples, on the basis of fraction total weight, in above-mentioned fraction, 3-O- galloyls -3,3',5,5', 7- penta hydroxy groups flavane or trans- -3-O- galloyls -3,3',5,5', 7- penta hydroxy groups flavane, its isomers, its is pharmaceutically acceptable Salt, its prodrug, its hydrate or its solvate can be 0.0001 weight % or more, 0.0005 weight % or more, 0.001 Weight % or more, 0.005 weight % or more, 0.01 weight % or more, 0.05 weight % or more, 0.1 weight % or more, 0.5 weight Measure % or more, 1 weight % or more, 1.5 weight % or more, 2 weight % or more, 2.5 weight % or more, 3 weight % or more, 3.5 Weight % or more, 4 weight % or more or 4.5 weight % or more or 5 weight % or less, 4.5 weight % or less, 4 weight % Below, 3.5 weight % or less, 3 weight % or less, 2.5 weight % or less, 2 weight % or less, 1.5 weight % or less, 1 weight Measure % or less, 0.5 weight % or less, 0.1 weight % or less, 0.05 weight % or less, 0.01 weight % or less, 0.005 weight Measure % or less, 0.001 weight % or less or 0.0005 weight % or less.0.0001 weight % to 5 weights can be preferably comprised Measure %.

According to other concrete examples, 3-O- galloyls -3,3&apos of the administration based on above-mentioned composition;,5,5', 7- penta hydroxy group flavane Or trans- -3-O- galloyls -3,3',5,5', 7- penta hydroxy groups flavane, its isomers, its pharmaceutically acceptable salt, its prodrug, its The dosage of hydrate or its solvate can be 0.0001g/kg/ days or more, 0.001g/kg/ days or more, 0.01g/kg/ Its above, 0.1g/kg/ days or more, 1g/kg/ days or more, 2g/kg/ days or more, 3g/kg/ days or more, 4g/kg/ days or more, 5g/ Kg/ days or more, 6g/kg/ days or more, 7g/kg/ days or more, 8g/kg/ days or more, 9g/kg/ days or more, 10g/kg/ days or more, 11g/kg/ days or more, 12g/kg/ days or more, 13g/kg/ days or more or 14g/kg/ days or more or 15g/kg/ days or less, 14g/kg/ days or less, 13g/kg/ days or less, 12g/kg/ days or less, 11g/kg/ days or less, 10g/kg/ days or less, 9g/kg/ Its following, 8g/kg/ days or less, 7g/kg/ days or less, 6g/kg/ days or less, 5g/kg/ days or less, 4g/kg/ days or less, 3g/ Kg/ days or less, 2g/kg/ days or less, 1g/kg/ days or less, 0.1g/kg/ days or less, 0.01g/kg/ days or less, 0.001g/kg/ It is following or 0.0005g/kg/ days or less.It can be preferably 0.0001g/kg/ days to 15mg/kg/ days.

According to other concrete examples, above-mentioned cognitive function lowly can be by being selected from the poly- of beta-amyloid protein (β-amyloid) Collect (aggregation), beta amyloid patch (plaque) is formed, brain-derived neurotrophic factor (brain-derived Neurotrophic factor, BDNF) expression is reduced and DNMT1 (DNA (cytimidine -5)-transmethylase 1) is expressed in increasing Any above cause.

According to another concrete example, above-mentioned cognitive function can include lowly selected from failure of memory, cognitive decline, identification energy One or more of power decline, depression and amnesia.

According to another concrete example, above-mentioned improvement can pass through β-shallow lake of aggregation inhibition, aggregation selected from beta-amyloid protein Decomposition, BDNF expression enhancing and the DNMT1 of powder sample albumen express one or more of reduction and realize.

An embodiment according to the present invention utilizes above-mentioned trans- -3-O- galloyls -3,3',5,5', the implementation of 7- penta hydroxy group flavane Beta-amyloid aggregation evaluate (beta-Amyloid aggregation assay) as a result, as comparison other substance What epicatechin 3-O- gallic acid esters (Epicatechin 3-O-gallate, ECG, 1 μ g/ml) were handled with unused compound Control group (positive control) is compared, and shows 17% aggregation inhibition, trans- -3-O- galloyls -3,3',5, 5', 7- penta hydroxy groups flavane compared with the control group, shows 64% aggregation inhibition.In particular, trans- galloyl-3-3-O-, 3',5,5', the beta-amyloid aggregation inhibition of 7- penta hydroxy group flavane is shown to be made in result chart (with reference to Fig. 5) For (phenol red, 55% aggregation inhibition) phenol red known to excellent agglutination inhibitor and morin (morin, 78% aggregation inhibition) between degree excellent aggregation inhibition.Moreover, in trans- -3-O- galloyls -3,3', 5,5', in 7- penta hydroxy group flavane processing groups, the reaction time shows the state of aggregation (aggregation) reduction after sixty minutes, It follows that trans- -3-O- galloyls -3,3',5,5', decomposition of the 7- penta hydroxy groups flavane also to the beta-amyloid protein of aggregation (disassemble) effectively.Therefore, it can confirm trans- -3-O- galloyls -3,3&apos using the present invention;,5,5', 7- penta hydroxy group flavane It can improve low with the relevant cognitive function of beta-amyloid protein.Caused by beta-amyloid aggregation furthermore, it is possible to confirm Disease (including the neurodegenerative diseases such as Alzheimer disease) prevention and treatment in can usefully use trans- -3-O- Galloyl -3,3',5,5', 7- penta hydroxy group flavane.

In addition, trans- -3-O- galloyls -3,3&apos of the present invention;,5,5', 7- penta hydroxy group flavane enhances BDNF in nerve cell Expression and the expression for reducing DNMT1.That is, found out the present application can be usefully applied to BDNF express reduce or DNMT1 expression enhances in the prevention and treatment of the neurodegenerative disease of low, Alzheimer disease of relevant cognitive function etc..

In another aspect of this invention, above-mentioned composition can be the drug for preventing or treating neurodegenerative disease Composition.

According to a concrete example, above-mentioned neurodegenerative disease can be by the aggregation of beta-amyloid protein (β-amyloid) (aggregation), during the patch (plaque) of beta-amyloid protein is formed, BDNF expression is reduced and DNMT1 expression increases Cause more than any.

According to other concrete examples, above-mentioned neurodegenerative disease can include Alzheimer disease, may be accompanied with include The low symptom of above-mentioned cognitive function.

Aforementioned pharmaceutical compositions according to an aspect of the present invention can by oral, non-oral, rectum, it is local, transdermal, Intravenously, intramuscular, intraperitoneal, subcutaneous etc. are administered.For oral medication dosage form can be tablet, pill, it is soft and Hard capsules, granule, powder, fine granule, liquor, opacifiers or micropill preparation, but not limited to this.For non-oral administration Dosage form can be solution, suspending agent, emulsion agent, gel, injection, point drops, suppository, patch or spray, but not It is defined in this.Above-mentioned dosage form can be easily manufactured by the usual way in the field, can also be included surfactant, be assigned Shape agent, hydrating agents, emulsification promoter, suspending agent, the salt for adjusting osmotic pressure or buffer, colorant, spice, stabilization Agent, preservative, preservative agent or other common auxiliary agents.

The dosage or dosage of aforementioned pharmaceutical compositions according to an aspect of the present invention can be according to pairs being administered The judgement of age, gender, weight, pathological state and its severity, administration route or prescriber of elephant and it is different.Based on this In the level of those skilled in the art, 1 day administration dosage of active ingredient for example can be for the determination of the dosage of kind factor 0.001 μ g/kg/ days to 5000mg/kg/ days, more specifically can be 0.0001mg/kg/ days to 15mg/kg/ days, but and unlimited Due to this.

According to another concrete example, above-mentioned composition can be food compositions.

The dosage form of above-mentioned food compositions is not particularly limited, but can for example be made such as tablet, granule, pill, powder The liquid agents such as last agent, potus;Caramel;Gel;Stick;The dosage forms such as tea bag.Those skilled in the art can according to dosage form or use mesh , without difficulty to ingredient usually used in the field in the food compositions of each dosage form other than active ingredient It is suitably selected and is coordinated, in the case where being applicable in simultaneously with other raw materials, it may occur however that synergy.

Above-mentioned composition can by simply taking in, drinking, drug administration by injection, spray delivery or squeeze administration etc. a variety of methods It is administered.

In food compositions according to an aspect of the present invention, the dosage of above-mentioned active ingredient is determined in this field skill In the level of art personnel, 1 day administration dosage for example can be 0.0001mg/kg/ days to 15mg/kg/ days, but be not limited to This, can be different according to many factors such as the age of the object of desired administration, health status, complication.

Food compositions according to an aspect of the present invention for example can be chewing gum, caramelization product, candy, ice cream class, The various foodstuffs such as Biscuits, the beverage products such as cold drink, mineral water, alcoholic beverage, including vitamin or minerals etc. Healthy food class.

In addition to the foregoing, food compositions according to an aspect of the present invention can include various nutritional agents, vitamin, The fragrance such as minerals (electrolyte), synthetic perfume and natural perfume material, colorant and thickener (cheese, chocolate etc.), pectic acid And its salt, alginic acid and its salt, organic acid, protective colloid tackifier, pH adjusting agent, stabilizer, preservative, glycerine, alcohol, use In the carbonating agent etc. of soda.In addition to this, food compositions according to an aspect of the present invention can also include and be used for Manufacture the pulp of fruit juice, fruit drink and vegetable beverage.This ingredient can be used alone or in combination.Although this addition The ratio of agent is not critically important, but under normal circumstances, the compositions according to an aspect of the present invention of every 100 parts by weight 0 to Include in the range of about 50 parts by weight.

The composition and effect of this specification are more specifically illustrated in the following, enumerating embodiment, experimental example and dosage form example. But these examples only with help understand this specification illustration purpose and be provided, the scope of this specification and Range is not limited to following examples.

<Embodiment 1>The manufacture of post-fermented tea sample

Water is added in the green tea made of green tea (Camellia sinensis var.Yabukita) leaf and by moisture Content is adjusted to 40 weight %.It is inoculated with 5 × 10 herein⁶The bacillus subtilis (Bacillus subtillis) of cfu/g, 50 DEG C fermentation 3 days after, fermented at 80 DEG C 4 days.It is 4 weights to be dried the tea of fermentation to moisture with 70 DEG C to 80 DEG C of hot winds After measuring % to 6 weight %, cured at 10 DEG C 100 days.

The tea sample of above-mentioned curing is crushed 15 seconds, is sieved with the stainless steel of mesh size 1mm.Then, by powder Broken 50mg is added in the microcentrifugal tube (Eppendorf tube) of 1.5ml, adds the deionized water of 1ml, in 60 DEG C of perseverances After being stirred 30 minutes with constant speed in warm water tank, centrifugation in 15 minutes has been carried out with 13000rpm at 25 DEG C.From drying Fermentation green-tea extract in only detached part not soluble in water.

<Embodiment 2>Fraction acquisition is detached with compound

The above-mentioned post-fermented tea sample of 150g is fractionated with acetone and is eliminated as much as catechin-derived object and coffee Cause obtains the soluble matter of flavonoids concentration.For the above-mentioned acetone soluble matter of 40g, using silica gel column chromatography, by chloroform:Methanol Mixture as solvent, with 10:1,5:1,1:The sequence of 1 (v/v), which is dissolved out, has obtained each fraction.

Herein, with high performance liquid chromatography (high performance liquid chromatography, HPLC, Waters Alliance 2695system, Waters, USA) to chloroform:Methanol 10:The knot that 1 (v/v) fraction 12g is analyzed Fruit is still caffeine and exists as main compound in the case where not being fractionated as described above with acetone, institute as above It states and confirms that caffeine has removed (Fig. 1) using after acetone fractionation.

Utilize high capacity highly effective adverse current chromatogram (high-performance countercurrent Chromatography, HPCCC, Dynamic Extractions Ltd, UK) it is fractionated the chloroform for eliminating caffeine of 8.9g: Methanol 10:1 (v/v) fraction.At this moment the solvent used is n-hexane-TBME (methyl tertiary butyl ether(MTBE), Methyl tert- Butyl ether)-BuOH-MeCN- water (0.25:3:1:1:5, v/v), flow velocity 25ml/min.Using above-mentioned condition, separate Totally 10 sub- fractions (Fig. 2), are analyzed, it is thus identified that the compound contained by each fraction is largely flavonoids with HPLC (flavonoid) system (Fig. 3, Fig. 4).

Trans- -3-O- galloyls -3,3&apos is isolated from above-mentioned fraction;,5,5', 7- penta hydroxy group flavane (Trans-3-O- galloyl-3,3',5,5', 7-pentahydroxyflavan, C₂₂H₁₈O₁₀, molecular weight 442.0900), the compound of separation Structure is as follows.

[Chemical formula 2]

¹In the case of H nuclear magnetic resonance (nuclear magnetic resonance, NMR), as solvent (solvent) methanol-d3 (methanol-d3) has been used, equipment has used Bruker Advance DPX-500, and (BRUKER is public Department, USA), trans- -3-O- galloyls -3,3',5,5', the NMR data of 7- penta hydroxy group flavane is as follows, specific chart such as Fig. 6.

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

<Experimental example 1>Beta-amyloid aggregation effect confirms

It confirmed above-mentioned trans- -3-O- galloyls -3,3&apos with fluorescence analysis (ThioflavinT assay);,5,5', 7- five The beta-amyloid aggregation of hydroxyl flavane hinders effect.

Specifically, beta-amyloid protein (A β 1-42, American Peptide Co, USA) is used with 5 μM of concentration, It is taken care of at -80 DEG C using preceding.Diluted respectively in DMSO morin (Morin, 100 μM), it is phenol red (Phenol Red, 100 μM), epicatechin 3-O- gallic acid esters (Epicatechin 3-O-gallate, ECG, 1 μ g/ml), trans- galloyl-3-3-O-, 3',5,5', 7- penta hydroxy groups flavane (1 μ g/ml) is adjusted to above-mentioned concentration.In order to determine that A β 1-42 assemble inhibition level, 92.5 It is separately added into each compound of 5 μ l prepared with above-mentioned concentration in the 0.01M buffer solution of sodium phosphate of μ l, the 5 of 2.5 μ l are added μM A β 1-42 after, after being reacted 26 hours at 37 DEG C, 5 μM of thioflavin Ts (ThioflavinT) of 2000 μ l are added, with fluorescence point Light analyzer (RF-5300PC, SHIMADZU CORPORATION, Japan) is measured.

The results are shown in Figure 5 for it.That is, ECG and trans- galloyl-3-3-O-, 3 ', 5,5 ', 7- penta hydroxy group flavane is relative to sun Property control group (positive control only assemble the group of beta-amyloid protein without compound processing) show respectively 17%, 64% aggregation inhibition.Especially trans- galloyl-3-3-O-, 3 ', 5, the inhibition of 5 ', 7- penta hydroxy group flavane (64%) phenol red (55%) and morin (78%) as positive inhibitor (positive inhibitor) are illustrated on Between degree excellent aggregation inhibition.In trans- galloyl-3-3-O-, 3 ', 5,5 ', 7- penta hydroxy group flavane processing groups In, assemble reduced state after sixty minutes by the reaction time and can be seen that trans- galloyl-3-3-O-, 3 ', 5,5 ', 7- penta hydroxy groups are yellow Alkane is also effective to the decomposition (disassemble) of the beta-amyloid protein of aggregation.

<Experimental example 2>Intracellular BDNF (brain-derived neurotrophic factor, brain-derived neurotrophic Factor) confirm with the expression quantity of DNMT1 (DNA (cytimidine -5)-transmethylase 1)

By SH-SY5Y (neuroblastoma, Korea Cell strain bank) cell strains 6 orifice plates (well plate, FALCON with every hole 2X10 on)⁶It is inoculated with (seeding), in 37 degree, 5%CO₂After being cultivated 24 hours in incubator, with 10 μM GCG, 10 μM of EGCG, existing green-tea extract (green tea extract, GTE) 10 μ g/ml, trans- galloyl-3-3-O-, 10 μ g/ml of 3 ', 5,5 ', 7- penta hydroxy group flavane, 5- -2 ' deoxycytidines of azepine (5-Aza-2 ' as positive controls Deoxycytidine) 1 μM of (5-Aza, Sigma-aldrich) is handled, and has further been cultivated 24 hours.Then, removal is complete Portion's culture medium is extracted RNA using RNA extracts kits (RNeasy mini kit, Quiagen companies).Utilize ultraviolet detection After device (TECAN companies) is quantitative to the mRNA of extraction, the mRNA of 1 μ g is utilized into kit (SuperScript VILO cDNA Synthesis Kit, Thermofisher scientific companies) synthesize complementary DNA.Take out the complementary DNA of about 1 μ g, profit It is carried out with Taqman probes (Life technology companies) and Quantitect probe PCRs kit (Quiagen companies) real When quantitative polyase chain reaction.The expression quantity of BDNF and DNMT1 are thereby confirmed that.At this moment, it as calibration standard mRNA, is utilized GAPDH as house-keeping gene (housekeeping gene).

The expression quantity of BDNF and DNMT1 is shown in Tables 1 and 2.

[Table 1]

The relative expression quantity of BDNF

It distinguishes	%
		Control group (no processing group)	100
Trans- galloyl-3-3-O-, 3 ', 5,5 ', 7- penta hydroxy groups flavane, 10 μ g/ml	123
		1 μM of -2 ' deoxycytidine of 5- azepines	149

[Table 2]

The relative expression quantity of DNMT1

It distinguishes	%
		Control group (no processing group)	100
Trans- galloyl-3-3-O-, 3 ', 5,5 ', 7- penta hydroxy groups flavane, 10 μ g/ml	81
		1 μM of -2 ' deoxycytidine of 5- azepines	65

<Dosage form example 1>Soft capsule

To according to embodiments of the present invention 2 post-fermented tea acetone fraction 150mg, lactose 440mg, cornstarch 430mg It is mixed with magnesium stearate 2mg and has manufactured soft capsule filling liquid.In addition, above-mentioned steps are different from, with 66 weight of gelatin Part, the ratios of 10 parts by weight of 24 parts by weight of glycerine and Sorbitol solution manufacture soft balsam capsule skin, above-mentioned filling liquid is filled, to manufacture Soft capsule.

In addition, to trans- galloyl-3-3-O- according to the ... of the embodiment of the present invention, 3 ', 5,5 ', 7- penta hydroxy group flavane 3mg, lactose 440mg, cornstarch 450mg and magnesium stearate 2mg are mixed to manufacture soft capsule filling liquid.Moreover, passing through above-mentioned side Formula manufactures soft capsule skin, above-mentioned filling liquid is filled, to manufacture soft capsule.

<Dosage form example 2>Tablet

To according to embodiments of the present invention 2 post-fermented tea acetone fraction 150mg, vitamin E 15mg, vitamin C 15mg, galactooligosaccharicomposition 250mg, lactose 60mg and maltose 140mg are mixed, and are granulated using fluid bed dryer Afterwards, it is added to sugar ester (sugar ester) 8mg.The composition is subjected to film-making with usual way and has manufactured tablet.

In addition, to trans- galloyl-3-3-O- of embodiment, 3 ', 5,5 ', 7- penta hydroxy group flavane 3mg, vitamin E 15mg, Vitamin C 15mg, galactooligosaccharicomposition 259mg, lactose 60mg and maltose 140mg are mixed, using fluid bed dryer into After row granulating, it is added to sugar ester (sugar ester) 8mg.The composition is subjected to film-making with conventional method and has manufactured piece Agent.

<Dosage form example 3>Potus

To according to embodiments of the present invention 2 post-fermented tea acetone fraction 80mg, vitamin E 9mg, vitamin C 9mg, After glucose 10g, citric acid 0.6g and liquid oligosaccharides 25g are mixed, the pure water that 400ml is added is filled.It is filled into After bottle, sterilizes 4~5 seconds at 30 DEG C and manufactured potus.

In addition, to trans- galloyl-3-3-O- of embodiment, 3 ', 5,5 ', 7- penta hydroxy group flavane 0.1mg, vitamin E 9mg, After vitamin C 9mg, glucose 10g, citric acid 0.6g and liquid oligosaccharides 25g are mixed, the pure water of 400ml is added, with The mode of every bottle of 200ml is filled.It is sterilized 4~5 seconds at 30 DEG C after being filled into bottle and has manufactured potus.

<Dosage form example 4>Granule

To according to embodiments of the present invention 2 post-fermented tea acetone fraction 150mg, vitamin E 9mg, vitamin C 9mg, Anhydrous crystal glucose 250mg and starch 550mg are mixed, and after being shaped to particle using fluid bed granulate machine, are filled into bag In and manufactured granule.

In addition, to trans- galloyl-3-3-O- of embodiment, 3 ', 5,5 ', 7- penta hydroxy group flavane 3mg, vitamin E 9mg, dimension Raw element C 9mg, anhydrous crystal glucose 250mg and starch 550mg are mixed, and particle is shaped to using fluid bed granulate machine Afterwards, it is filled into bag and has manufactured granule.

<Dosage form example 5>Injection

Composition recorded in following Table 3 has manufactured injection with conventional method.

[Table 3]

Gradation composition	Content
		According to the post-fermented tea acetone fraction of embodiment 2	100mg
Injection sterile purified water	In right amount
		PH adjusting agent	In right amount

In addition, trans- galloyl-3-3-O- of the embodiment using 3mg, 3 ', 5,5 ', 7- penta hydroxy group flavane replace it is above-mentioned after Fermented tea acetone fraction and manufactured injection.

<Dosage form example 6>Healthy food

According to the composition described in following table 4, healthy food has been manufactured with conventional method.

[Table 4]

In addition, trans- galloyl-3-3-O- of the embodiment using 3mg, 3 ', 5,5 ', 7- penta hydroxy group flavane replace it is above-mentioned after Fermented tea acetone fraction, to manufacture healthy food.

About the ratio of components of said vitamin and mineral mixture, to be relatively suitble to the ingredient of healthy food as an example And composition is mixed, but its match ratio can be implemented with random variation, be mixed according to conventional healthy food manufacturing method above-mentioned After ingredient, it can be used in the manufacture of health food composition according to the conventional method.

<Dosage form example 7>Healthy beverage

[Table 5]

As shown in Table 5 above, the pure water for adding surplus makes total volume become 900ml, according to conventional healthy beverage After manufacturing method mixes mentioned component, after 85 DEG C of agitating and heatings about 1 hour, manufactured solution is filtered, is taken to 2 liters of sterilizing In container, after sealing sterilizing, refrigeration keeping, to manufacture healthy beverage.

In addition, trans- galloyl-3-3-O- of the embodiment using 0.1mg, 3 ', 5,5 ', 7- penta hydroxy group flavane replaces above-mentioned Post-fermented tea acetone fraction, to manufacture healthy beverage.

The specific part of this specification is illustrated in detail above, to those skilled in the art, this The specific technology of kind is preferred embodiment, and it is obvious that the range of this specification, which will not be defined in this,.Therefore, this explanation The substantial range of book is based on appended claims and its equivalent is defined.

Claims

1. a kind of low improvement composition of cognitive function, contains the 3-O- galloyls -3,3&apos indicated by following chemical formula 1;,5, 5', 7- penta hydroxy groups flavane, its isomers, its pharmaceutically acceptable salt, its prodrug, its hydrate, its solvate or comprising Its post-fermented tea ketone fraction as active ingredient,

Chemical formula 1

2. composition according to claim 1, wherein the post-fermented tea ketone fraction is for the non-aqueous of post-fermented tea The ketone fraction of soluble extract.

3. composition according to claim 1, wherein the ketone is acetone.

4. composition according to claim 1, wherein the 3-O- galloyls -3,3',5,5', 7- penta hydroxy group flavane be by Trans- -3-O- galloyls-the 3,3&apos that following chemical formula 2 indicates;,5,5', 7- penta hydroxy group flavane,

Chemical formula 2

5. composition according to claim 1, wherein in the composition, 3-O- galloyls -3,3',5,5', 7- pentahydroxy-s Base flavane, its isomers, its pharmaceutically acceptable salt, its prodrug, its hydrate or its solvate content relative to institute It is 0.000001 weight % to 5 weight % to state composition total weight.

6. composition according to claim 1, wherein in the composition, the content phase of post-fermented tea ketone fraction It is 0.01 weight % to 100 weight % for the composition total weight.

7. composition according to claim 1, wherein in the fraction, the 3-O- galloyls -3,3',5,5',7- Penta hydroxy group flavane, its isomers, its pharmaceutically acceptable salt, its prodrug, its hydrate or its solvate are total with fraction Include 0.0001 weight % to 5 weight % on the basis of weight.

8. composition according to claim 1, wherein 3-O- galloyls -3,3&apos of the administration based on the composition;,5, 5', 7- penta hydroxy groups flavane, its isomers, its pharmaceutically acceptable salt, its prodrug, its hydrate or its solvate are given Dose is 0.0001mg/kg/ days to 15mg/kg/ days.

9. composition according to any one of claim 1 to 8, wherein the cognitive function is lowly by being selected from β-shallow lake The aggregation of powder sample albumen, BDNF expression reduces and DNMT1, that is, DNA (cytimidine -5)-transmethylase Caused by 1 expression increases.

10. composition according to any one of claim 1 to 8, wherein the cognitive function is lowly comprising selected from memory One or more of power decline, cognitive decline, recognition capability decline, depression and amnesia.

11. composition according to any one of claim 1 to 8, wherein the improvement is by being selected from beta-amyloid protein Aggregation inhibit, aggregation beta-amyloid protein decomposition, BDNF expression enhancing and DNMT1 expression reduce one or more of and It carries out.

12. composition according to any one of claim 1 to 8, wherein the composition is for preventing or treating god Pharmaceutical composition through degenerative disease.

13. composition according to claim 12 is pharmaceutical composition, wherein the neurodegenerative disease is by selecting It is expressed caused by one or more of reduction and DNMT1 expression increases from the aggregation of beta-amyloid protein, BDNF.

14. composition according to claim 12 is pharmaceutical composition, wherein the neurodegenerative disease is A Er Ci Haimo diseases.

15. composition according to any one of claim 1 to 8, wherein the composition is food compositions.