JP2007320958A - Preventive and / or therapeutic agent for fatty liver - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a preventive and / or therapeutic agent for naturally occurring fatty liver.
The preventive and / or therapeutic agent for fatty liver according to the present invention is characterized by comprising, as an active ingredient, a high molecular weight polyphenol extracted from fermented tea such as oolong tea or black tea.
[Selection] Figure 2

Description

  The present invention relates to a prophylactic and / or therapeutic agent for naturally occurring fatty liver.

Fatty liver is a condition in which a large amount of lipid accumulates in hepatocytes due to excessive intake of alcohol, overnutrition, diabetes, etc., and it can cause chronic hepatitis, cirrhosis, liver cancer, etc. It is important. Therefore, research and development of drugs effective for the prevention and treatment of fatty liver are energetically performed. For example, Patent Document 1 discloses that an angiotensin converting enzyme inhibitor is effective for the prevention and treatment of fatty liver. Are listed.
JP 2003-63990 A

However, the search for a safe drug that has an excellent effect on fatty liver is significant.
Accordingly, an object of the present invention is to provide a preventive and / or therapeutic agent for naturally occurring fatty liver.

  As a result of intensive studies in view of the above points, the present inventors have found that a polymer polyphenol extracted from fermented tea is effective for the prevention and treatment of fatty liver.

The preventive and / or therapeutic agent for fatty liver of the present invention based on the above findings is characterized in that, as described in claim 1, high molecular polyphenol extracted from fermented tea is an active ingredient.
Further, the preventive and / or therapeutic agent according to claim 2 is the prophylactic and / or therapeutic agent according to claim 1, wherein the polymer polyphenol has a partial structure having a structure in which a procyanidin structure and B rings of catechins are bonded to each other. The number average molecular weight is 9000 to 18,000.
Further, the preventive and / or therapeutic agent according to claim 3 is the prophylactic and / or therapeutic agent according to claim 1 or 2, wherein the polymer polyphenol is extracted by extracting the water-eluting component in the fermented tea leaves with ethyl acetate. The non-eluting component of ethyl acetate that has not been extracted is extracted with butanol, and the extracted butanol-eluting component is obtained by fractional purification using water-containing acetone as a solvent.
Further, the preventive and / or therapeutic agent according to claim 4 is the same as the prophylactic and / or therapeutic agent according to claim 1 or 2, wherein the polymer polyphenol is extracted by extracting the water-eluting component in the fermented tea leaves with ethyl acetate. Extract the non-eluting components of ethyl acetate with butanol, acidify the non-eluting components of butanol that were not extracted, and then extract with butanol again, and fractionate and purify the extracted butanol-eluting components using aqueous acetone as a solvent. It is characterized by being obtained.
The preventive and / or therapeutic agent according to claim 5 is characterized in that in the prophylactic and / or therapeutic agent according to any one of claims 1 to 3, the fermented tea is oolong tea or black tea.

  According to the present invention, a prophylactic and / or therapeutic agent for naturally occurring fatty liver can be provided.

The preventive and / or therapeutic agent for fatty liver according to the present invention is characterized by comprising a high-molecular polyphenol extracted from fermented tea as an active ingredient. In the present invention, fermented tea means tea obtained by causing fermentation of tea leaves, and specific examples thereof include oolong tea that is semi-fermented tea and black tea that is fermented tea. In the present invention, the high molecular polyphenol is a compound in which a compound having a plurality of phenolic hydroxyl groups in the molecule, for example, catechins that are flavonoid polyphenols are highly polymerized (for example, the number average molecular weight is 5000 to 30000). means. Catechins are a group of compounds having a flavan-3-ol skeleton having a plurality of phenolic hydroxyl groups in the C ₆ -C ₃ -C ₆ skeleton, and are particularly well known to those skilled in the art that they are contained in large amounts in tea leaves. It is the fact that was done. The chemical structure of catechin is as shown below. There are various derivatives of catechin (for example, galcatechin in which hydrogen at the 5′-position of catechin is substituted with a hydroxyl group, and galcatechin gallate in which the hydroxyl group at the 3-position is ester-bonded with gallic acid) . In the present invention, catechins mean catechin and derivatives thereof, and specific examples thereof include catechin, catechin gallate, epicatechin, epicatechin gallate, gallocatechin, gallocatechin gallate, epigallocatechin gallate, etc. Is mentioned.

Polymer polyphenols extracted from fermented tea are thought to be produced by highly polymerizing catechins in the production process of fermented tea (fermentation, ripening, etc.). As an acquisition method thereof, for example, fermented tea leaves Extraction of water-eluted components (components eluted by extraction with water, boiling water, hot water, steam, etc.) with ethyl acetate, extracted non-extracted ethyl acetate with butanol, and extracted with butanol A method of fractionating and purifying components using water-containing acetone as a solvent, and extracting water-eluted components in fermented tea leaves with ethyl acetate, extracting non-extracted ethyl acetate with butanol, and extracting non-butanol with non-extracted butanol Examples include a method in which butanol is extracted again after acidifying the components, and the extracted butanol-eluting components are fractionated and purified using aqueous acetone as a solvent. . In addition, fractional purification using water-containing acetone as a solvent can be performed, for example, by gel filtration chromatography using Toyopearl HW-40F manufactured by Tosoh Corporation.
In addition, high-molecular polyphenols extracted from fermented tea can be obtained by converting fermented tea leaf extracts obtained by extraction operations using water, organic solvents such as methanol, ethanol, and acetone into synthetic adsorbents (for example, aromatic synthetic adsorbents). Adsorbed on a certain Mitsubishi Chemical Corporation Diaion HP20, etc., and after washing the adsorbent with a 30% methanol solution, etc., the adsorbed component was eluted with a 70% methanol solution, etc., and the ethyl acetate eluting component was removed from the eluted component. Subsequently, the ethyl acetate non-eluting component can also be obtained by a method of fractional purification by gel filtration chromatography using hydrous acetone as a solvent.

  Specific examples of the high molecular weight polyphenol extracted from fermented tea include a procyanidin structure and a structure in which B rings of catechins are bonded to each other in a partial structure, and the number average molecular weight is 9000 to 18000. Can be mentioned. The procyanidin structure is a structure in which the C ring of catechins and the A ring of other catechins are bonded. Polymer polyphenols extracted from fermented tea include a procyanidin structure and a structure in which the B rings of catechins are bonded to each other, as well as a structure in which the B ring of catechins and the A ring of other catechins are bonded. May be. Specific examples of the partial structure of the polymer polyphenol extracted from fermented tea include the following. The partial structures shown below are merely examples, and the polymer polyphenol extracted from fermented tea is considered to have a wide variety of partial structures including partial structures including benzotropolone, benzoquinone, naphthoquinone, and the like. It is done.

  The preventive and / or therapeutic agent for fatty liver according to the present invention is prepared by formulating a high molecular polyphenol extracted from fermented tea into an oral preparation such as a granule, a tablet or a capsule by a method known per se, or a non-injection such as an injection. When formulated into oral preparations and administered to mammals including humans, it exhibits excellent effects in the prevention and treatment of fatty liver. The dose can be appropriately determined depending on the age, sex, weight, physical condition, degree of fatty liver, etc. of the application target. Moreover, the preventive and / or therapeutic agent for fatty liver of the present invention is effective enough to bring high-molecular polyphenols extracted from fermented tea to various forms of food (including supplements) for the prevention and treatment of fatty liver. You may eat as a functional food by adding quantity.

  EXAMPLES Hereinafter, although an Example demonstrates this invention in detail, this invention is not limited at all by the following description.

Example 1: Action of high-molecular polyphenol extracted from oolong tea (part 1) on fatty liver (A) Preparation of high-molecular polyphenol extracted from oolong tea 30 g of oolong tea leaf was added to 1000 mL of boiling water and boiled for about 1 minute. Allowed to stand for 10 minutes. Thereafter, the oolong tea leaves were removed by filtration to obtain a filtrate. The above operation was performed 4 times in total to obtain an aqueous solution containing a water-eluting component from 120 g of oolong tea leaves.
Next, extraction with ethyl acetate was performed to remove low molecular weight polyphenols from the water-eluting components. Specifically, 200 mL of water-saturated ethyl acetate was added to 500 mL of an aqueous solution, stirred and allowed to stand, and then the operation of separating the ethyl acetate phase was repeated 10 times to obtain an aqueous phase containing an ethyl acetate non-eluting component. .
Next, the aqueous phase containing non-eluting components of ethyl acetate was extracted with butanol. Specifically, after removing the remaining ethyl acetate by concentrating the aqueous phase under reduced pressure, 200 mL of water-saturated n-butanol was added per 500 mL, stirred, allowed to stand, and then fractionating the butanol phase. Repeated 10 times. By collecting the separated butanol phase, an extract containing a butanol-eluting component was obtained, and this was concentrated under reduced pressure to remove the remaining butanol, thereby obtaining an aqueous solution containing a butanol-eluting component. This aqueous solution was freeze-dried, and a neutral fraction extracted from oolong tea butanol was obtained at a yield of 4.5 g per 120 g of oolong tea leaves.

Next, the oolong tea butanol-extracted neutral fraction obtained by the above method was further finely fractionated by gel filtration chromatography. Toyopearl HW-40F (manufactured by Tosoh Corporation) was used as the stationary phase, and water-containing acetone was used as the mobile phase.
First, Toyopearl HW-40F was packed in a column having a diameter of 2.4 cm and a length of 35 cm. In addition, 600 mL each of a 20% acetone solution and a 50% acetone solution were prepared as mobile phases.
Next, 0.3 g of a neutral fraction extracted from oolong tea butanol was dissolved in 3 mL of 20% acetone solution, and the resulting solution was applied to a column. The components adsorbed on the stationary phase using a 20% acetone solution and a 50% acetone solution are sequentially eluted at a rate of 0.3 g / min with a linear concentration gradient of 20 to 50%, and an eluate is obtained using a fraction collector. Was dispensed into test tubes in 5-g increments.
Next, the absorbance at 350 nm of the eluate fractionated into each test tube was measured to prepare an elution curve. The neutral fraction extracted from oolong tea butanol was further finely fractionated based on the prepared elution curve, and after collecting the eluate belonging to the same fraction, acetone was removed by concentration under reduced pressure, freeze-dried, and 15 Fraction samples were obtained.
FIG. 1 shows an elution curve (elution pattern) of a neutral fraction extracted from oolong tea butanol (horizontal axis: test tube number from which the eluate was collected (in order of collection), vertical axis: absorbance at 350 nm). 15 fractions obtained by subfractionation based on the elution curve are also shown in FIG.

(B) Measurement of the average molecular weight of the fraction (15) The average molecular weight of the fraction (15) was measured by a size exclusion chromatography method (SEC: size exclusion chromatography). An LC-10A system (manufactured by Shimadzu Corporation) was used as a high-speed chromatograph, and TSK-GELα-3000 (column dimensions: diameter 7.8 mm × length 30 cm, manufactured by Tosoh Corporation) was used as a column. The column temperature was 40 ° C. Dimethylformamide containing 10 mM lithium chloride was used as the developing solvent. The flow rate was set at 0.6 mL / min. The detector used was a UV detector included in the LC-10A system. The detection wavelength was set at 275 nm.
First, TSK standard polystyrene (manufactured by Tosoh Corporation) is applied to the column as a molecular weight standard compound, and the elution time is plotted on the horizontal axis and the UV detection value is plotted on the vertical axis to create an elution curve, and calibration is performed based on this. A curve was created.
Next, a fraction (15) dissolved in dimethylformamide was applied to the column, the elution time was plotted on the horizontal axis, the UV detection value on the vertical axis, and the average molecular weight was calculated based on the calibration curve. The average molecular weight was 1.52 × 10 ⁴ and the weight average molecular weight was 2.10 × 10 ⁴ .

(C) Structural analysis of fraction (15) Structural analysis of fraction (15) was performed using a pyrolysis-gas chromatograph-mass spectrum (Py-GC-MS) analyzer. According to the Py-GC-MS analyzer, after pyrolyzing a sample with a pyrolyzer (Py), the obtained pyrolyzed product is introduced into a gas chromatograph (GC) and separated, and further separated substances Can be obtained with a mass spectrum apparatus (MS) to obtain knowledge about the thermal properties and chemical structure of the sample.
First, the fraction (15) was pyrolyzed using a Curie point pyrolyzer JHP-5 (manufactured by Nippon Analytical Industrial Co., Ltd.) as the pyrolyzer (Py). First, the temperature in the furnace and the gas chromatograph introduction part was set to 250 ° C. Next, 0.1 to 0.2 mg of the fraction (15) is wrapped with ferromagnetic-pyrofoil (thickness 50 μm), 5 μL of 10% tetramethylammonium hydroxide methanol solution is added, and placed in a furnace at 315 ° C. Was subjected to pyrolysis by treating for 4 seconds, and then the pyrolysis product was led to a gas chromatograph. The methanol solution of 10% tetramethylammonium hydroxide is used for the purpose of obtaining volatility and thermal stability at the stage of mass spectrometry by methylating the chemical structure contained in fraction (15). It was.
A gas chromatograph mass spectrometer JMS-600M (manufactured by JEOL Ltd.) was used for gas chromatograph-mass spectrum (GC-MS). In addition, TSS-2000 (manufactured by Nippon Analysis Co., Ltd.) was used as a data processing device. A capillary column HP-1MS (column size: diameter 0.25 mm × length 30 m, coated liquid layer thickness: 0.25 μm, manufactured by Agilent Technologies) was used as the column for gas chromatography. The pyrolysis product of fraction (15) and the carrier gas were introduced into the column to separate the substances present in the pyrolysis product and to acquire data on the retention time. Moreover, mass spectrometry was performed about each isolate | separated substance, and the knowledge regarding chemical structure etc. was acquired. The temperature in the column was first maintained at 50 ° C. for 1 minute, then linearly increased to 300 ° C. at 5 ° C./minute, and then maintained at 300 ° C. for 14 minutes. Helium was used as the carrier gas. The flow rate was set at 1 mL / min. Mass spectrometry was performed under the conditions of an ion source temperature of 250 ° C. and an ionization voltage of 70 eV.
As a result of comparing the data obtained by gas chromatograph-mass spectrum and the data obtained by conducting the same experiment on the synthetic standard substance, the following 10 chemical formulas were obtained from the pyrolysis product of fraction (15). Of the compound was detected.

In addition, the retention time (tR: retention time) and molecular weight of said compound are as follows.
Compound 1 ... tR: 23.0 minutes, molecular weight: 168
Compound 2 ... tR: 22.1 min, molecular weight: 166
Compound 3 tR: 30.3 minutes, molecular weight: 196
Compound 4 ... tR: 30.5 minutes, molecular weight: 226
Compound 5 ... tR: 31.1 minutes, molecular weight: 254
Compound 6 ... tR: 32.4 minutes, molecular weight: 254
Compound 7 ... tR: 32.9 minutes, molecular weight: 254
Compound 8 ... tR: 35.2 minutes, molecular weight: 284
Compound 9 ... tR: 36.9 minutes, molecular weight: 312
Compound 10 ·· tR: 46.5 minutes, molecular weight: 450

  From the above results, it was found that the fraction (15) is a substance having a chemical structure that gives the 10 kinds of pyrolysis products. From the results of mass spectrometry, this substance is located between the 2′-position, 5′-position and 6′-position of the catechins and any of the other catechins, for example, the 8th and 6′-positions of the catechins. It was suggested that there are polymerization sites between the 6-position and the 6'-position, between the 6'-position and the 6'-position. In addition, this substance contains a gallic acid residue that is considered to be ester-bonded to the hydroxyl group at the 3-position of epicatechin or epigallocatechin from the result of tannase degradation of fraction (15) performed separately. It was shown that. From the result of hydrochloric acid-butanol decomposition, it was speculated that this substance contained a procyanidin structure polymerized between the 4-position and 8-position or between the 4-position and 6-position of catechins in the partial structure. Comprehensively judging the above results, the fraction (15) is, for example, a polymer polyphenol that contains at least a procyanidin structure and a structure in which B rings of catechins are bonded to each other in the partial structure shown below. It was found that there are various types of partial structures contained in this polymer polyphenol, and the above results do not deny the existence of partial structures containing benzotropolone, benzoquinone, naphthoquinone, etc. Absent).

(D) Action of high molecular weight polyphenol of fraction (15) on liver fat (experimental method)
A male 6-week-old type II diabetes model mouse (BSK.Cg-m + / + Lepr <db> / Jcl, Claire, Japan) was treated with a high molecular weight polyphenol dissolved in physiological saline at a dose of 0.09 mg / animal. Each day, 0.1 mL was injected intraperitoneally. After 10 weeks from the start of administration, the mice were treated with Nembutal, and then whole blood was collected. The liver was cut into an appropriate size, fixed with a 3.7% formalin solution, dehydrated with a 70% ethanol solution, dehydrated while changing the butanol concentration, and then embedded in paraffin. The paraffin-embedded liver was cut into a thickness of 8 μm with a microtome and placed on a slide glass to obtain a liver tissue section. After staining liver tissue sections with hematoxylin for staining nuclei and eosin for staining cytoplasm, images were obtained using an optical microscope. The obtained image is shown in FIG. Further, FIG. 3 shows an image of a liver tissue section 10 weeks after the start of administration of a control mouse in which physiological saline was injected intraperitoneally at 0.1 mL / animal.

(Experimental result)
As is clear from FIG. 2 and FIG. 3, the fat contained in the liver tissue of the mouse administered with the high molecular weight polyphenol of fraction (15) is compared with the fat contained in the liver tissue of the control mouse administered with physiological saline. There was much less (a fine outline represents fat. A large outline is a blood vessel). Moreover, the degree of weight gain of the mice administered with the high molecular weight polyphenol of fraction (15) was not significantly different from the degree of weight gain of the control mice, and progressed smoothly. From the above results, it was found that the polymer polyphenol of the fraction (15) is an excellent therapeutic agent for fatty liver caused by type II diabetes.

Example 2: Effect of macromolecular polyphenol extracted from oolong tea (part 2) on fatty liver The aqueous phase remaining after obtaining the butanol-eluting component in Example 1 (A) was adjusted to about pH with hydrochloric acid. Then, 200 mL of water-saturated n-butanol was added to 500 mL, and the mixture was stirred and allowed to stand, and then the butanol phase was separated five times. By collecting the separated butanol phase, an extract containing a butanol-eluting component was obtained, and this was concentrated under reduced pressure to remove the remaining butanol, thereby obtaining an aqueous solution containing a butanol-eluting component. This aqueous solution was freeze-dried to obtain an acidic fraction extracted from oolong tea butanol with a yield of 3.2 g per 120 g of oolong tea leaves. This oolong tea butanol-extracted acidic fraction was finely fractionated in the same manner as in Example 1 (A) to obtain 15 fraction samples. FIG. 4 shows an elution curve (elution pattern) of an acidic fraction extracted from oolong tea butanol (horizontal axis: test tube number from which the eluate was collected (in order of collection), vertical axis: absorbance at 350 nm). 15 fractions obtained by subfractionation based on the elution curve are also shown in FIG. When the average molecular weight of the high molecular weight polyphenol of the fraction (14) was measured by the size exclusion chromatography method in the same manner as in Example 1 (B), the number average molecular weight was 1.73 × 10 ⁴ and the weight average molecular weight was 2. .44 × 10 ⁴ . When the action of this fraction (14) on the fatty liver of the polymer polyphenol was evaluated in the same manner as in Example 1 (D), the same action as that of the polymer polyphenol in fraction (15) of Example 1 was observed. It was.

Example 3: Effect of high molecular polyphenol extracted from black tea (part 1) on fatty liver Add 25 g of black tea leaf to 500 mL of boiling water, boil gently for 10 minutes, and immediately filter and remove black tea leaf with Buchner funnel The filtrate was obtained. The above operation was performed 4 times in total to obtain an aqueous solution containing a water-eluting component from 100 g of black tea leaves. The subsequent operation was carried out in the same manner as in Example 1 (A), and a black fraction extracted with black tea butanol was obtained at a yield of 1.5 g per 100 g of black tea leaves. This black butanol-extracted neutral fraction was finely fractionated in the same manner as in Example 1 (A) to obtain 16 fraction samples. FIG. 5 shows the elution curve (elution pattern) of the neutral fraction extracted from black tea butanol (horizontal axis: test tube number from which the eluate was collected (in order of collection), vertical axis: absorbance at 350 nm). In addition, 16 fractions obtained by subfractionation based on the elution curve are shown in FIG. When the average molecular weight of the high molecular weight polyphenol of the fraction (15) was measured by the size exclusion chromatography method in the same manner as in Example 1 (B), the number average molecular weight was 1.36 × 10 ⁴ and the weight average molecular weight was 1. It was 89 × 10 ⁴ . When the effect of this fraction (15) on the fatty liver of the polymer polyphenol was evaluated in the same manner as in Example 1 (D), the effect similar to that of the polymer polyphenol in fraction (15) of Example 1 was observed. It was.

Example 4: Effect of high molecular polyphenol extracted from black tea (part 2) on fatty liver In the same manner as in Example 2, an acidic fraction extracted from black tea butanol was obtained in a yield of 1.9 g per 100 g of black tea leaves. The black tea butanol-extracted acidic fraction was finely fractionated in the same manner as in Example 1 (A) to obtain 11 fraction samples. FIG. 6 shows an elution curve (elution pattern) of the acidic fraction extracted from black tea butanol (horizontal axis: test tube number from which the eluate was collected (in order of collection), vertical axis: absorbance at 350 nm). In addition, 11 fractions obtained by subfractionation based on the elution curve are shown in FIG. When the average molecular weight of the high molecular weight polyphenol of the fraction (11) was measured by the size exclusion chromatography method in the same manner as in Example 1 (B), the number average molecular weight was 9.43 × 10 ³ and the weight average molecular weight was 1. 48 × 10 ⁴ . When the action of this fraction (11) on the fatty liver of the polymer polyphenol was evaluated in the same manner as in Example 1 (D), the same action as that of the polymer polyphenol in fraction (15) of Example 1 was observed. It was.

Example 5: Effect of Oral Administration of Fraction (15) of Example 1 on Fatty Liver Male 6-week-old Type II Diabetes Model Mouse (BSK.Cg-m + / + Lepr <db> / Jcl, Nippon Claire) was allowed to freely drink water containing a high-molecular polyphenol of fraction (15) of Example 1 at a concentration of 0.02% for 10 weeks. Also, water containing epigallocatechin gallate at a concentration of 0.02% and water containing nothing were allowed to freely drink for 10 weeks respectively. After 10 weeks, the effect of each mouse on fatty liver was evaluated in the same manner as in Example 1 (D). The results are shown in FIG. In the figure, (a) is a mouse administered with water containing nothing, (b) is a mouse administered with water containing the polymer polyphenol of fraction (15) of Example 1, and (c) is epigallocatechin gallate. The result of the mouse | mouth which administered water containing is shown. As is clear from FIG. 7, the fat contained in the liver tissue of the mouse administered with water containing epigallocatechin gallate was almost the same as the fat contained in the fat tissue of the mouse administered with water containing nothing. However, the fat contained in the liver tissue of the mouse administered with water containing the high molecular weight polyphenol of fraction (15) of Example 1 was compared with the fat contained in the fat tissue of the mouse administered with water containing nothing. There was much less (a fine white outline represents fat. A large white outline is a blood vessel).
In addition, after 10 weeks, the mice administered with water containing the polymer polyphenol of fraction (15) of Example 1, the mice administered with water containing epigallocatechin gallate, and the mice administered with water containing nothing. Various items were measured for each (n = 5-6). The results are as follows. The mice administered with water containing the high molecular weight polyphenol of fraction (15) of Example 1 were significantly suppressed in weight gain and blood glucose level compared to mice administered with water containing nothing, Such a phenomenon was not observed in mice administered with water containing epigallocatechin gallate. Although the mice administered with water containing the high molecular weight polyphenol of fraction (15) of Example 1 had significantly higher food intake compared to mice administered with water containing nothing, visceral fat Although the amount was significantly lower, such a phenomenon was not seen in mice administered water containing epigallocatechin gallate. Mice administered with water containing the polymeric polyphenol of fraction (15) of Example 1 have significantly lower plasma total cholesterol levels and free fatty acid levels than mice administered with water containing nothing. The level of triglycerides was also low, although there was no significant difference, but this phenomenon was not observed in mice administered with water containing epigallocatechin gallate.
From the above results, the high-molecular polyphenol of fraction (15) of Example 1 itself or itself is degraded in the intestine, and then taken into the body through the intestinal tract, causing liver dysfunction and the like. Not found to improve fatty liver.

Formulation Example 1: Tablet 5 g of lyophilized powder of high molecular weight polyphenol of fraction (15) of Example 1, 78 g of lactose and 17 g of magnesium stearate were uniformly mixed to obtain tablets according to a conventional method.

Formulation Example 2: Biscuit Soft flour 31 g, whole egg 16 g, butter 16 g, sugar 24 g, water 10 g, baking powder 1 g, fraction 3 of Example 3 (15) high-molecular polyphenol lyophilized powder 2 g, 100 g in total. Biscuits were made according to the law.

  The present invention has industrial applicability in that a prophylactic and / or therapeutic agent for naturally occurring fatty liver can be provided.

It is a graph which shows 15 fractions obtained by subfractionating based on the elution curve of the oolong tea butanol extraction neutral fraction in Example 1, and an elution curve. It is an optical microscope photograph which shows the effect | action of the high molecular polyphenol of the fraction (15) with respect to the fatty liver of a type II diabetes model mouse. It is an optical micrograph which shows the effect | action of the physiological saline similarly. It is a graph which shows 15 fractions obtained by subfractionating based on the elution curve of the oolong tea butanol extraction acidic fraction in Example 2, and an elution curve. It is a graph which shows 16 fractions obtained by subfractionating based on the elution curve of the black butanol extraction neutral fraction in Example 3, and an elution curve. It is a graph which shows eleven fractions obtained by subfractionating based on the elution curve of the black tea butanol extraction acidic fraction in Example 4, and an elution curve. 2 is an optical micrograph showing the effect of oral administration of the polymer polyphenol of fraction (15) of Example 1 on fatty liver of type II diabetes model mice in Example 5. FIG.

Claims (5)

  A preventive and / or therapeutic agent for fatty liver, characterized by comprising a high-molecular polyphenol extracted from fermented tea as an active ingredient.   2. The prevention according to claim 1, wherein the high-molecular polyphenol contains at least a procyanidin structure and a structure in which B rings of catechins are bonded to each other, and has a number average molecular weight of 9000 to 18000. / Or therapeutic agent.   High molecular weight polyphenol extracts water-eluting components in fermented tea leaves with ethyl acetate, non-extracted ethyl acetate non-eluting components are extracted with butanol, and the extracted butanol-eluting components are fractionated and purified using water-containing acetone as a solvent. The preventive and / or therapeutic agent according to claim 1 or 2, wherein the prophylactic and / or therapeutic agent is obtained.   High molecular weight polyphenols extract the water-eluted components in the fermented tea leaves with ethyl acetate, extract the non-extracted ethyl acetate non-eluted components with butanol, acidify the non-extracted butanol non-eluted components and extract again with butanol, The preventive and / or therapeutic agent according to claim 1 or 2, wherein the extracted butanol-eluting component is obtained by fractional purification using water-containing acetone as a solvent.   The preventive and / or therapeutic agent according to any one of claims 1 to 3, wherein the fermented tea is oolong tea or black tea.