JP5794414B2 - Pharmaceutical composition - Google Patents

Description

The present invention relates to a pharmaceutical composition containing a compound having a specific structure, which is produced by fermentation of tea leaves using black koji and obtained by extraction and purification.
In particular, the present invention relates to a pharmaceutical composition that is effective in suppressing blood sugar level elevation, reducing triglycerides, suppressing blood pressure elevation, improving cholesterol metabolism, improving liver and kidney function, improving brain function, and preventing and treating Alzheimer's disease. Belongs to the pharmaceutical preparation technology.

Tea as a luxury product has been used habitually all over the world for a long time, but most of them are pouring hot water into tea leaves and drinking the exudate.
Since the exudate contains various components extracted from tea leaves, it has a unique aroma and taste and has a specific function.
For this reason, research on the components of tea leaves has been conducted extensively, and the following is also known regarding the components.

That is, the characteristic component of tea is caffeine and a tannin-based substance catechin, and caffeine has an excitatory effect on humans, has a bitter taste, and also has a diuretic effect.
This catechin is the most abundant tea ingredient and is a tea astringent ingredient.
Besides these, amino acids typified by theanine, vitamins typified by vitamin C, chlorophylls, inorganic components such as potassium and calcium, and fragrance components such as dimethyl sulfide, green leaf alcohol, and terpene alcohol are widely known. .

It is also known that these components change at the stage of tea production from tea leaves.
For example, Kama fried tea and roasted green tea are known to contain a large amount of nitrogen-containing compounds such as pyrazine and pyrrole, and fermented tea is known to contain a large amount of terpene alcohol with floral aroma (published by Heibonsha: World Encyclopedia reference).

As described above, since tea leaves contain a lot of various useful components, many attempts have been made since long ago to extract the components efficiently and to process the tea leaves to obtain many active ingredients.
In particular, in regard to fermented tea, for example, Japanese Patent Application Laid-Open No. 2002-370994 (Patent Document 1) discloses black blood sugar, that is, blood sugar obtained by hot water extraction from black tea aged by a post-fermentation method using black koji mold or the like. It has been reported that value-inhibiting substances have been obtained.

  Furthermore, JP-A-2005-341876 (Patent Document 2) reports that tea having a maltase inhibitory effect can be obtained by adding koji mold or koji to tea leaves and fermenting them.

On the other hand, the inventors of this application previously filed a patent application by finding that a medicinal composition capable of preventing deterioration of liver function and the like can be obtained from fermented tea leaves by extraction with acetic acid.
The contents are disclosed in Japanese Patent Application Laid-Open No. 2007-238584 (Patent Document 3).

Furthermore, following the previous research, the inventors pursued the characteristics of various ingredients of fermented tea (post-fermented tea) in more detail, and examined the preparation conditions of fermented tea, changes in the ingredients contained by the raw tea leaves, etc. did.
Furthermore, in order to make more effective use of widely used tea leaves, the study also examined changes in the active ingredients in tea leaves and their fermentation conditions, and filed a patent application for the results, the contents of which are described in JP2010-220489A. No. 4 (Patent Document 4).

JP 2002-370994 A (Claims) Japanese Patent Laying-Open No. 2005-341876 (Claims) JP 2007-238584 A (Claims) JP 2010-220489 A (Claims)

  The inventors identified the X component (presumed to be a gallic acid-derived compound) that was found to be produced by fermentation of tea leaves using black koji in the previous study, and the pharmacology was particularly effective. Pursuing more deeply about the characteristics, we studied to further utilize tea leaves, especially to make effective use of the found X component.

As a result, the inventors identified an X component produced in tea leaves fermented with black koji, and found that this X component also has pharmacological properties that could not be found in the previous stage. Completed the invention.

That is, the invention according to claim 1 of the present invention is
Characterized in that it contains a compound represented by the following formula 1, improving liver function, it is a pharmaceutical composition for use in improving cholesterol metabolism improvement or renal function.

The pharmaceutical composition according to the present invention contains a compound having a specific structure represented by the above formula 1, thereby suppressing an increase in blood glucose level, a reduction in neutral fat, an improvement in liver function, an increase in blood pressure, an improvement in cholesterol metabolism, or It is effective for improving renal function, improving brain function, preventing and treating Alzheimer's disease , especially improving liver function, improving cholesterol metabolism, or improving kidney function . The elucidation will allow the chemical synthesis of this compound and will allow a wide range of drug preparations in the pharmaceutical industry.

Since the compound in this invention can be used in a powder state or as acetic acid or water or an ethanol solution, it is effective and effective when used as a pharmaceutical agent in order to exhibit the above effects. It can be used in the future and is applied to health foods and has excellent effects.

It is a figure which shows the result of the HPLC analysis of the 4.5% acetic acid extract of fermented tea. It is an ultraviolet absorption spectrum of each peak of FIG. It is a fractional drawing by Bio-Gel column of 4.5% acetic acid extract of fermented tea. It is a fractional drawing by the Bio-Gel column of the ethyl acetate extract from fermented tea acetic acid extract. It is a HPLC chart of 3 fractions of FIG. FIG. 4 is a diagram of adsorption chromatography using a Sepdex LH column (2 × 27 cm) in fraction 5 of FIG. 4 is an HPLC chart of the fraction containing the X component of fraction 5. It is a HPLC chart of X component refine | purified by HPLC. It is an ultraviolet absorption spectrum of X component refine | purified by HPLC. It is a crystal photograph of X component. It is a figure which shows transition of the blood glucose level at any time in vivo using STZ mouse. It is a figure which shows transition of the alkaline phosphatase in vivo using STZ mouse | mouth. It is a figure which shows transition of the triglyceride in vivo using STZ mouse. It is a figure which shows transition of urea nitrogen in vivo using STZ mouse. It is a figure which shows transition of potassium in vivo using STZ mouse. It is a figure which shows transition of the crawl in vivo using a STZ mouse | mouth. It is a figure which shows transition of the blood glucose level at any time by the in vivo using a KK-Ay mouse | mouth. It is a figure which shows transition of the blood pressure at any time in vivo using a SHR mouse | mouth. It is a figure which shows transition of ALT by the in vivo using a SHR mouse | mouth. It is a figure which shows transition of HDL-cho / T-cho in vivo using a SHR mouse. It is a figure which shows transition of the crawl in vivo using a SHR mouse | mouth. It is a figure which shows the change of the average body weight in vivo using a ZDF rat. It is a figure which shows the body weight change rate with respect to the total intake in vivo using a ZDF rat.

  The specific compound represented by Formula 1 (hereinafter referred to as “specific compound”) used in the present invention is a fermented tea (hereinafter referred to as “fermentation”) obtained by fermentation of tea leaves using a conventionally known black rice cake. "Black tea"), which is produced by extraction and separation from fermented black tea, but since the structure has been elucidated, it can also be prepared by chemical synthesis.

As a tea leaf as a raw material used when obtaining this specific compound from fermented black tea,
Gyokuro, most tea, two flush, three flush, Fall flush the like, Ru is selected particularly bis crop.
As conditions for fermentation of these tea leaves, in general, conditions for preparing fermented tea using black koji, such as raw black koji, temperature during fermentation (usually 30 to 45 ° C.), moisture during fermentation (usually 25 to 35%), etc. Accordingly, the desired gallic acid-derived compound can be obtained.
In addition, the extraction of a specific compound from fermented black tea is also performed by a normal extraction method using dilute acetic acid, water, or hot water as an extraction medium.

  In the fermentation method itself of tea leaves in fermented black tea that produces the specific compound, there is no exceptional new condition, but in order to produce the specific compound and maintain the content after production, the following means are used. It is desirable to adopt.

As the fermentation condition for maintaining the state in which the fermented black tea contains the specific compound, the most important condition is the fermentation period.
Producing fermented black tea is said to take months if it is long.
Also in Patent Document 2, it is preferably about 3 weeks (paragraph 0030), but as a period for satisfying the above conditions, a preferable fermentation period for this invention is 3 to 10 days.
Moreover, aerobic fermentation is employ | adopted for fermentation.

  The specific compound thus obtained can be orally administered to a patient as a drug dissolved or suspended in a diluent mixed with a pharmaceutically acceptable carrier.

The pharmaceutical composition of this invention comprises suitable additives,
For example, lactose, sucrose, mannitol, corn starch, synthetic or natural gum, excipients such as crystalline cellulose, starch, cellulose derivatives, binders such as gum arabic, gelatin, polyvinylpyrrolidone, carboxymethyl cellulose cellulose, Disintegrants such as sodium carboxymethyl cellulose, starch, corn starch, sodium alginate, lubricants such as talc, magnesium stearate, sodium stearate, fillers or dilutions such as calcium carbonate, sodium carbonate, calcium phosphate, sodium phosphate A tablet, powder (powder), pill, granule or the like can be prepared by appropriately mixing with an agent.
Moreover, it can also be set as a capsule using a hard or soft gelatin capsule.

Further, the pharmaceutical composition of the present invention is dissolved in a generally used inert diluent such as purified water, and if necessary, this solution is infiltrated, emulsifier, dispersion aid, surfactant, sweetener. Liquid preparations such as syrups and elixirs can also be obtained by appropriately adding ingredients, flavors, fragrances and the like.
These preparations can be made by a conventionally known method.

When the pharmaceutical composition of the present invention is administered as a medicine to humans, the dosage is determined by the dosage of the specific compound as the active ingredient.
The dose depends on the dosage form, the age of the patient, etc., but is in the range of 1 mg to 1,000 mg per day, and is preferably 10 mg to 500 mg per day for an adult weighing 50 kg.

Hereinafter, based on an Example, the pharmaceutical composition of this invention is demonstrated in detail.
<Preparation of fermented black tea>
80 kg of raw tea leaves (raw tea) are put into a drum-type automatic koji making apparatus, and 80 kg of water is added with stirring by rotating the drum to swell the raw tea leaves, and then separately from the raw tea leaves (powdered tea) and black rice cake The prepared seed meal was added so as to be 10 ⁷ / g of Aspergillus niger (23 kg as the seed meal amount), and fermented at a set temperature of 35 ° C. for 6 days.
The temperature rise during fermentation was suppressed using an air cooling fan, and efforts were made to maintain the set temperature. The fermented tea leaves were sterilized by steaming at a temperature of 100 ° C. and dried at a temperature of 55 ° C. to obtain fermented black tea.

<Extraction and purification of specific compounds>
Extraction and purification of a specific compound (hereinafter referred to as “component X”) was carried out according to the following procedure.
First, 200 g of fermented black tea was used, extracted with 4.5% acetic acid as the extraction medium, and 72.6 g of extract (yield: 36.3%: hereinafter, black tea extraction) by concentration under reduced pressure and lyophilization. I got a thing).
FIG. 1 shows the results of HPLC analysis of this black tea extract, and FIG. 2 shows the ultraviolet absorption spectrum of each peak.
Peaks a to c in FIG. 1 are EGC, Caffeine, and EC, respectively, from the elution position and absorption spectrum, and peak d is the X component.

The extract was fractionated using a Bio-GelP-10 column (2 × 28 cm) with a 1% aqueous acetic acid solution as a developing solution, and the results are shown in FIG.
As a result of analyzing each fraction by HPLC, it was found that the X component was contained in fraction 5 together with the brown catechin polymer.

When fraction 5 containing the X component obtained by the above fractionation was extracted with ethyl acetate, it was confirmed that the X component was extracted into the ethyl acetate layer and the catechin polymer remained in the aqueous layer.
After concentrating the X component-containing ethyl acetate, the solution diluted with deionized water was applied to a Bio-GelP-10 column (3 × 30 cm) and developed with a 1% aqueous acetic acid solution. Shown in
As a result of performing the HPLC analysis about the fractions 4-6, it turned out that all contain X component, as shown in FIG.

The supernatant obtained by adding methanol to lyophilized fraction 5, stirring and centrifuging was applied to a Sephadex LH column (2 × 27 cm) equilibrated with methanol in advance and developed with methanol.
The elution pattern is shown in FIG. 6, and the HPLC pattern of the fraction having absorption at 300 nm is shown in FIG.

When this fraction was purified by HPLC of TFA-MeCN system, X component was obtained as purified crystals as shown in FIG.
The ultraviolet absorption spectrum was as shown in FIG.
Further, this fraction was dried under reduced pressure, deionized water was added, and the mixture was warmed to dissolve and then cooled. As a result, a crystalline gallic acid-derived compound as shown in FIG. 10 was obtained.
This compound is described in J. Org. Nat. Med. Epigallocatechin gallate disclosed in 15 Feb 2011; EGCG and gallocatechin gallate; the same as the compound biosynthesized from GCG, and had the structure represented by Formula 1.

The evaluation result of the function regarding the pharmacological properties of the obtained X component is as follows.
<Evaluation of antitoxic effect (in vivo experiment using STZ-induced rats)>
In order to evaluate the antitoxic effect of the X component in an in vivo experiment, 7-week-old Wistar rats were obtained from KBT Oriental Co., Ltd., and were grouped into 5 groups as shown in Table 2 below after 1 week of preliminary breeding. .
After preliminary breeding, streptozotocin (STZ) was intraperitoneally administered to groups 2 to 5 to produce STZ-induced type 1 diabetes model rats.
This Among 3-5 group was 3 weeks free feeding of solid shaped feed MF containing X component (black tea extract) (manufactured by Oriental Yeast) until sacrifice (12 weeks old).
The 1,2 group were given ad libitum only solid shaped feed MF.
The administration samples of each group and their intake amounts are as shown in Table 2 below.
In addition, about the compounding quantity of X component in this feed, it set so that it might become 1 time, 4 times, and 7 times the intake (66 mg / day) in a human clinical test.
The rats are housed in a conventional breeding room (lighting time from 7:00 to 19:00) at a room temperature of 24 ± 3 ° C. and a relative humidity of 55 ± 15% throughout the preliminary breeding period and experimental period, and the food intake and body weight are measured every week. In addition, the blood glucose level was measured before and after the sample administration, and at the first and second weeks after the sample administration.
Blood was collected from the heart at 12 weeks of age after sample administration for 3 weeks, and erythrocyte deformability measurement and blood biochemical tests (35 items such as cholesterol and blood glucose) were performed.

Blood was collected from the tail vein before STZ / sample administration (8 weeks old), 1 week after sample administration, and 2 weeks after sample administration.
Whole blood was measured with a simple blood glucose meter.
Statistical processing was carried out between the two groups of each age group, 3, 4, and 5 groups, and the results are shown in FIG.
Marks in the figure indicate that there is a significant difference (*; p <0.05, **; p <0.01).

As shown in the transition of blood glucose level at any time in FIG. 11, the blood glucose level of the diabetic control group (group 2) increases significantly as the age of the week increases as compared to the normal group (group 1).
In contrast, groups 3 and 5 to which the X component was administered showed significantly lower values than group 2 at the first week of STZ administration.
Also in Group 4, the increase in blood glucose level almost reached its peak at the second week of sample administration, and was significantly lower than that in Group 2.
This suggests that the X component suppresses an increase in blood glucose level, although the time of suppression differs depending on the sample concentration.

<Blood chemistry test>
Three weeks after administration of STZ (12 weeks of age), laparotomy was performed under isoflurane (Dainippon Pharmaceutical) anesthesia, and blood was collected from the heart.
Fasting was started 18 hours before slaughter.
CBC measurement (Sysmex) was performed on the collected blood, and biochemical examination was performed on the obtained plasma (2000 rpm, 10 minutes, temperature 25 ° C.).
The results are shown in FIG. 12 (alkaline phosphatase), FIG. 13 (neutral fat), FIG. 14 (urea nitrogen), FIG. 15 (potassium), and FIG.
Statistical processing is carried out between the two groups of each age group, 3, 4, and 5 groups, and the marks in the figure indicate that there is a significant difference / trend (#; p <0.08 *; p <0.05). **; p <0.01).

As is clear from the above figure, in the blood biochemical test, there was a significant difference or tendency between the 2 groups and the X component administration group for 5 items of alkaline phosphatase, neutral fat, urea nitrogen, potassium and chlor. .
First, alkaline phosphatase, which is an index of liver function, is significantly higher in group 2 than in group 1, and the function of liver function is reduced by STZ administration.
On the other hand, the 4 groups administered with the X component showed a tendency to be lower than the 2 groups.
There was no significant difference / trend in the other 3 and 5 groups, but both showed lower values than the 2 groups.
This suggested that the X component tends to suppress or improve the decrease in liver function.
About neutral fat, the 4th and 5th group showed the significantly low value compared with the 2nd group.
This suggests that the X component suppresses the increase in neutral fat caused by STZ administration.
Regarding urea nitrogen, which is an index of renal function, the group 3 showed a tendency toward a lower value compared to the group 2.
Moreover, although there was no significant difference and a tendency also in the 4th and 5th groups, it became a value lower than the 2nd group, and the urea nitrogen value became low as the sample concentration became high.
This suggested that the X component may improve renal function in a concentration-dependent manner.
It was suggested that potassium and chlor, which are indicators of renal function other than urea nitrogen, improve or tend to improve renal function in a concentration-dependent manner.
Concentration-dependent results were seen for all three indicators of renal function.

<Evaluation of antidiabetic effect (in vivo experiment with KK-Ay mouse)>
In order to evaluate the anti-metabolic syndrome effect of the X component in an in vivo experiment, KK-Ay / Taj mouse ♂ 3 weeks old (CLEA Japan, Inc.) was obtained from Kudo, Inc. As shown in FIG.
Sterile tap water was given until the first week of pre-breeding, and sterile tap water with 0.05% ascorbic acid was given for the second week.
The sample concentration was calculated from the daily intake of the X component (black tea extract) administration experiment (administration by ingestion) conducted using the same model last year.
In addition, about the compounding quantity of X component in this sample, it set based on the previous clinical trial result.
Also, all administration is free water consumption were given ad libitum solid shaped feed MF (manufactured by Oriental Yeast) to feed.
The mice are bred in a conventional breeding room (lighting time from 7:00 to 19:00) at a room temperature of 24 ± 3 ° C. and a relative humidity of 55 ± 15% throughout the preliminary breeding period and experimental period, and the amount of food intake and body weight is measured every week. did.
Furthermore, the blood glucose level at the time of sacrifice at 4 weeks, 5 weeks, 7 weeks after sample administration was measured.
After sample administration for 7 weeks (at the age of 12 weeks), blood was collected from the heart and subjected to blood biochemical tests (34 items such as cholesterol and blood glucose).

<Changes in blood glucose level as needed>
Blood samples were collected from the tail vein at the time of sacrifice at 4th, 5th and 7th weeks after sample administration.
Fasted for 5 to 6 hours before blood collection to obtain fasting blood glucose.
The result of measuring whole blood with a simple blood glucose meter is shown in FIG.
Marks in the figure indicate that there is a significant difference (*; p <0.05 **; p <0.01, ***; p <0.001).

Normally, KK-Ay mice develop hyperglycemia from 7 to 8 weeks of age.
There is no significant difference between groups 1 and 2 after 4 weeks of sample administration (at 9 weeks of age), but 2 groups are significantly lower after 5 weeks of sample administration (at 10 weeks of age). It was.
This suggested that the X component markedly suppresses the increase in blood glucose level in diabetes model mice.
In addition, since the effect in a model that genetically develops type 2 diabetes has been proved this time, prevention of diabetes in humans (blue boys) who have a predisposition to type 2 diabetes due to the X component is expected.

<Evaluation of antihypertensive effect (in vivo experiment using SHR rat)>
In order to evaluate the antihypertensive effect of the X component (black tea extract) in an in vivo experiment, a 7-week-old spontaneously hypertensive rat (SHR rat) was obtained from KBT Oriental Co., Ltd.
After preliminary breeding for 1 week, the groups were divided as shown in Table 4 so that the average blood pressure was equivalent.
Sample administration was started at 9 weeks of age after preliminary breeding.
One group solid form feed MF (manufactured by Oriental Yeast), and the two groups was 4 weeks free feeding the solid shaped feed MF containing X component.
The amount of intake in group 2 is as shown in Table 3.
In addition, about the compounding quantity of X component (black tea extract) in this feed, it set so that it might become 7 times the intake (66 mg / day) in a human clinical trial based on the result of a prior clinical trial.
The rats are housed in a conventional breeding room (lighting time from 7:00 to 19:00) at a room temperature of 24 ± 3 ° C. and a relative humidity of 55 ± 15% throughout the preliminary breeding period and the experimental period. It was measured.
In addition, the blood pressure was measured before the sample administration and at the first week, the second week, the third week, and the fourth week (before sacrifice) after the sample administration.
Blood was collected from the abdominal aorta at the age of 13 weeks after sample administration for 4 weeks, and blood biochemical tests (34 items such as cholesterol and blood glucose) were performed.

<Changes in blood pressure as needed>
The blood pressure was measured for a total of five systolic blood pressures (SBP) before the sample administration, and after the sample administration, 1 week, 2 weeks, 3 weeks, and 4 weeks (before sacrifice).
The measurement was performed using a non-invasive blood pressure measuring device (Softlon Co., Ltd.).
The measurement was performed at least three times, and the average value of the three times was taken as the blood pressure of the individual, and the results are shown in FIG.
Marks in the figure indicate that there is a significant difference / trend (#; p <0.08 *; p <0.05, **; p <0.01).

As shown in the figure, in the sample non-administered group (Group 1), the blood pressure increases as the age increases.
On the other hand, in the sample administration group (group 2), although it increases, the onset speed of hypertension is slower than that in group 1.
In the 2nd week of sample administration, the 2 group tended to be lower than the 1 group, and the 3rd and 4th weeks were significantly lower.
From this, it was shown that X component (black tea extract) suppresses a blood pressure rise notably.

<Blood biochemistry test>
Four weeks after administration of the sample (age 13 weeks), the abdomen was opened under isoflurane (Dainippon Pharmaceutical) anesthesia, and blood was collected from the abdominal aorta.
Fasting was started 18 hours before slaughter.
FIGS. 19 to 21 show the results of CBC measurement (Sysmex) on the collected blood, and biochemical tests performed on the obtained plasma (2,000 rpm, 10 minutes, temperature 25 ° C.).
Marks in the figure indicate that there is a significant difference / trend (#; p <0.08 *; p <0.05, **; p <0.01).

In the blood biochemistry test, significant differences or trends were observed for ALT, HDL-cho / T-cho, and crawl.
First, ALT, which is an index of liver function, showed a tendency for 2 groups to be lower than 1 group.
This suggested that the X component (black tea extract) may suppress or improve the decrease in liver function.
Next, in the item of HDL-cho / T-cho, 2 groups showed significantly higher values than 1 group.
Therefore, it was suggested that the black tea extract suppresses or improves the decrease in cholesterol metabolism.
Regarding chlor (Cl), which is an index of renal function, the two groups showed significantly high values.
Thereby, it can be said that X component has a function fall inhibitory effect or an improvement effect also in a renal function.

<Evaluation of anti-aging effect (in vivo experiment with ZDF rat)>
The anti-aging effect of the X component (black tea extract) In order to evaluate in vivo experiments, the obesity type 2 diabetes model rats ZDF 9 weeks of age, the solid shaped feed MF containing 0.65% Black tea extract (Oriental yeast) was fed freely for 4 weeks.
The control group was given ad libitum only solid shaped feed MF.
In addition, the average intake amount of the fermented tea extract from this feed corresponds to about 10 times the intake amount (66 mg / day) in the human clinical trial.
The rats are housed in a conventional breeding room (lighting time from 7:00 to 19:00) at a room temperature of 24 ± 3 ° C. and a relative humidity of 55 ± 15% throughout the preliminary breeding period and the experimental period. It was measured.

The average body weight after 12 weeks of breeding is shown in FIG. 22, and the rate of weight change relative to the total intake is shown in FIG.
As is clear from these figures, the sample containing the X component (black tea extract) is less likely to gain weight even if the same calorie equivalent is consumed compared to the control sample not containing it. Even if it has characteristics, it strongly suggests that the X component (black tea extract) suppresses the weight gain of rats and is effective in preventing metabolic syndrome.

<Acute oral toxicity test (animal safety test)>
For the X component, an acute oral toxicity test using rats was requested from the Japan Food Analysis Center.
The test group was administered with a sample at a dose of 150 mg / kg, and the control group was orally administered with water for injection as a solvent control to male and female rats once and observed for 14 days.
As a result, no abnormalities or deaths were observed during the observation period.
From this, it was considered that the LD50 value by single oral administration in the rat of the sample was 150 mg / kg or more in both sexes.

<Reversion mutation test>
For the black tea extract containing the X component, a reverse mutation test was requested from the Japan Food Analysis Center.
The test method uses Escherichia coli WP2uvrA and four Salmonella typhimurium TA strains in accordance with the attached “Genotoxicity Test Guidelines” of “Guidelines on Genotoxicity Tests of Drugs” (November 1, 1999, Pharmaceutical Trial No. 1604). Was done.
The sample dose was 313-5,000 μg / plate.
As a result, an increase in the number of revertant colonies was not observed.
From the above, it was concluded that the reverse mutagenicity of the specimen under the test conditions was negative.

The compound of the present invention specified by the above formula 1 suppresses an increase in blood glucose level, reduces neutral fat, improves liver function, suppresses an increase in blood pressure, improves cholesterol metabolism or kidney function, improves brain function, prevents Alzheimer's disease It is effective for treatment and not only enables further utilization of tea leaves, but also enables chemical synthesis of this compound by elucidating the structure.

  Since the pharmaceutical composition containing the specific compound according to the present invention has excellent pharmacological properties, the specific compound can be supplied as a non-toxic solvent solution of powder or water, acetic acid or ethanol. It is highly likely to be widely used in the tea manufacturing industry and the pharmaceutical manufacturing industry where this specific compound can be supplied.

Claims (1)

Characterized in that it contains a compound represented by the following formula 1, improving liver function, pharmaceutical compositions for use in improving cholesterol metabolism improvement or renal function.