JP2015017043A - Cholesterol absorption control agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a cholesterol absorption control agent containing a Ganoderma lucidum extract.SOLUTION: A Ganoderma lucidum extract shows an excellent cholesterol absorption control action, and more particularly, a preventing or improving action of arteriosclerosis, a promotion action of cholesterol elimination, and an uptake inhibitory action of cholesterol to lipid micelles. Therefore, it can utilize as medicaments, quasi drugs or foods having a safe and excellent cholesterol absorption control action.

Description

The present invention relates to a cholesterol absorption inhibitor characterized by containing a ganoderma extract.

Cholesterol is an important lipid as a constituent of animal cell membranes and as a precursor of bile acids, steroid hormones, vitamin D and the like. However, hypercholesterolemia in which the blood cholesterol level is high is considered to be the main cause of arteriosclerosis, and there is a risk of causing fatal diseases such as ischemic heart disease and cerebral infarction.

Examples of cholesterol in blood include bad cholesterol (low density lipoprotein, hereinafter referred to as LDL) and good cholesterol (high density lipoprotein, hereinafter referred to as HDL). When excess LDL is oxidized and accumulated, it adheres to the blood vessel wall and causes arteriosclerosis. On the other hand, HDL is considered to be an anti-arteriosclerotic factor because it functions to return excess cholesterol to the liver. That is, the ratio of LDL to HDL in the blood is important, and reducing the LDL / HDL ratio has attracted attention as a new index for preventing or improving arteriosclerosis. As a method of reducing the LDL / HDL ratio, reducing the ratio of LDL / HDL cholesterol concentration using defatted soybean has been proposed (Patent Document 1).

Moreover, as a conventional therapeutic agent for hypercholesterolemia, a method using a cholesterol synthesis inhibitor, an anion exchange resin preparation, or a cholesterol oxidation inhibitor is generally used. However, since side effects due to long-term continuous use of these drugs have also been reported, a method with a lower risk of side effects, for example, a method of suppressing absorption of dietary cholesterol and promoting excretion is desirable.

Conventional cholesterol absorption inhibitors include cholesterol absorption inhibitors containing bifidobacteria bacteria as an active ingredient (Patent Document 2), cholesterol absorption inhibition compositions by gel filtration fraction of whey ultrafiltration concentrate (Patent Document) 3) is known. Moreover, as a thing which accelerates | stimulates the excretion of cholesterol, the cholesterol discharge promoter (patent document 4) which uses the saponins obtained from tea as an active ingredient is known.

By the way, in order to absorb diet-derived cholesterol in the intestinal tract, a process in which cholesterol is taken into particles called lipid micelles formed by the action of bile is essential. Therefore, if cholesterol uptake into lipid micelles is suppressed, absorption of cholesterol in the intestinal tract can be suppressed. Conventionally, tea saponin compounds (Patent Document 5), specific lactic acid bacteria (Patent Document 6), royal jelly-derived proteins (Patent Document 7), and the like are known as those that suppress cholesterol uptake into lipid micelles.

Ganoderma is a basidiomycete belonging to the genus Ganodermaaceae and Ganoderma, and has been known as an immortal spirit since ancient times. And ganoderma (Akashiba), black ganoderma (black turf), purple ganoderma (purple turf), blue ganoderma (Aobashi), yellow ganoderma (yellow turf) and white ganoderma (white turf). ing.

As an effect of Ganoderma lucidum on cholesterol, an action of reducing blood cholesterol in beagle dogs (Non-patent Document 1) and rats (Non-patent Document 2) has been reported. However, Non-Patent Document 1 and Non-Patent Document 2 only describe that 1% Ganoderma was administered subcutaneously or intragastricly for 2 to 5 weeks. It is not described as to whether it is used or whether it is using Ganoderma itself or an extract extracted with some solvent, and the weight The dose per unit is not described, and detailed experimental methods and results are unknown. In addition, Non-Patent Documents 1 and 2 do not describe the LDL / HDL ratio lowering action, cholesterol excretion promoting action, cholesterol cholesterol uptake inhibiting action of cholesterol by Ganoderma extract, There is no description at all about the fact that cholesterol absorption suppression action is further enhanced by fractionating Ganoderma hot water extract with an organic solvent.

Further, an LDL receptor synthesis promoter (Patent Document 8) characterized by containing an extract of black ganoderma as a blood cholesterol lowering agent by ganoderma has been reported. However, the blood cholesterol lowering action described in Patent Document 8 is based on the promotion of LDL receptor synthesis in the liver. It suppresses cholesterol absorption, lowers the LDL / HDL ratio, promotes cholesterol excretion, and cholesterol micelles of cholesterol. No description has been made on the uptake-inhibiting action.
JP 2009-280517 A Japanese Patent No. 5116473 Japanese Patent No. 2935837 JP 2006-206472 A Japanese Patent No. 4828135 Japanese Patent No. 4662443 JP 2012-106988 A JP 2010-208965 A Small Animal Clinic 14 (3): 19-24, 1995 Small Animal Clinic 14 (4): 13-19, 1995

The object of the present invention relates to a cholesterol absorption inhibitor characterized by containing a ganoderma extract. More specifically, the present invention relates to a preventive or ameliorating agent for arteriosclerosis, a cholesterol excretion promoter, and an inhibitor of cholesterol uptake into lipid micelles, characterized by containing a ganoderma extract. The present invention also relates to a cholesterol absorption inhibitor characterized by containing an organic solvent-soluble fraction of Ganoderma hot water extract.

As a result of diligent research, the present inventors have found that Ganoderma extract has a cholesterol absorption inhibitory effect, an action to prevent or ameliorate arteriosclerosis, a cholesterol excretion promoting action, and an action to inhibit cholesterol uptake into lipid micelles. . Moreover, it discovered that an effect | action increased remarkably by changing an extraction solvent from water to lower alcohols, such as ethanol. Furthermore, the present inventors have found that the effect is remarkably enhanced by an organic solvent-soluble fraction obtained by extracting a ganoderma hot water extract with an organic solvent, and completed the present invention.

The ganoderma used in the present invention is a mushroom belonging to the genus Bamboo fungus, Ganodermaaceae, Ganoderma, Ganoderma (red turf), Black ganoderma (black turf), Purple ganoderma (purple turf), Blue ganoderma (Aoshiba), Yellow Reishi (Koshiba) and White Reishi (Shiroshiba) are known. In the present invention, all mushrooms belonging to the family Amanitaceae can be used, preferably ganoderma, black ganoderma, purple ganoderma, blue ganoderma, yellow ganoderma, white ganoderma, and more preferably ganoderma. It is desirable to use (Akashiba, Ganoderma lucidum).

Ganoderma used in the present invention can be used regardless of fruiting bodies, mycelium, natural products, cultivated products, and cultured products, and those widely distributed in the Chinese and Japanese markets can be used. . Moreover, the state as it is, a crushed material, a dried material, etc. can be selected suitably as needed, and it can attach to extraction operation.

Examples of the extraction solvent for obtaining the ganoderma extract used in the present invention include water, lower alcohols (methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, etc.), liquid polyhydric alcohols ( 1,3-butylene glycol, propylene glycol, glycerin, etc.), ketones (acetone, methyl ethyl ketone, etc.), acetonitrile, esters (ethyl acetate, butyl acetate, etc.), hydrocarbons (hexane, heptane, petroleum ether, etc.), ether (Ethyl ether, tetrahydrofuran, propyl ether, etc.). These solvents may be used alone or in combination of two or more. Preferably, water, ethanol, and a mixture thereof are desirable.

Examples of a method for further fractionating the Ganoderma hot water extract of the present invention to obtain an organic solvent soluble fraction of the Ganoderma hot water extract include, for example, lower alcohols (methanol, ethanol, 1-propanol, 2-propanol). , 1-butanol, 2-butanol, etc.), liquid polyhydric alcohols (1,3-butylene glycol, propylene glycol, glycerin, etc.), ketones (acetone, methyl ethyl ketone, etc.), acetonitrile, esters (ethyl acetate, butyl acetate, etc.) ), Hydrocarbons (hexane, heptane, petroleum ether, etc.), ethers (ethyl ether, tetrahydrofuran, propyl ether, etc.). These solvents may be used alone or in combination of two or more. Butanol and ethyl acetate are preferable.

The intake amount of the organic solvent soluble fraction of the ganoderma extract used in the present invention and the ganoderma hot water extract varies depending on the administration form, purpose of use, age, body weight, etc., but it is 1 to 20000 mg in terms of dry matter. / Day, preferably 10 to 10000 mg / day, more preferably 50 to 5000 mg / day. An amount smaller than the above dosage range may be sufficient, or it may be necessary to administer beyond the range. In addition, the method for adding medicinal ingredients in the formulation may be added in advance or during production, and may be appropriately selected in consideration of workability.

The cholesterol absorption inhibitor of the present invention can be used for any of foods, quasi-drugs or pharmaceuticals. Examples of the dosage form include powders, granules, tablets, sugar-coated tablets, capsules for oral use, Syrups, pills, suspensions, solutions, emulsions, etc. It can be an injection for parenteral use. It can also be a suppository.

As for the cholesterol absorption inhibitor of the present invention, the organic solvent soluble fraction of Ganoderma extract or Ganoderma hot water extract may be used as it is, as long as the effect of the extract is not impaired, as necessary. Excipients, stabilizers, preservatives, binders, disintegrants, hydrocarbons, fatty acids, alcohols, esters, pH adjusters, preservatives, fragrances used in ordinary pharmaceuticals, quasi drugs or foods Etc. can also be blended.

The cholesterol absorption inhibitor characterized by containing the ganoderma extract of the present invention has an action of suppressing cholesterol absorption, an effect of preventing or improving arteriosclerosis, an action of promoting cholesterol excretion, and an action of suppressing the incorporation of cholesterol into lipid micelles. Have. The cholesterol absorption inhibitor of the present invention is excellent in safety because it is extracted from a natural material with experience of eating.

Production Example 1 Ganoderma Hot Water Extract 400 g of purified water was added to 20 g of dried ganoderma (Ganoderma lucidum), extracted at 95-100 ° C. for 2 hours, filtered, and the filtrate was concentrated and frozen. After drying, 1.4 g of Ganoderma hot water extract was obtained.

Production Example 2 Ganoderma Hot Water Extract
400 g of purified water is added to 20 g of the dried product of black ganoderma and extracted at 95-100 ° C. for 2 hours, followed by filtration. The filtrate is concentrated and freeze-dried to obtain 1.1 g of ganoderma hot water extract. It was.

Production Example 3 Ganoderma Ethanol Extract 900 g of ethanol was added to 100 g of dried ganoderma (Ganoderma lucidum), extracted at room temperature for 7 days, filtered, and the filtrate was concentrated to dryness. 1.6 g of extract was obtained.

Production Example 4 Ganoderma Ethanol Extract
900 g of ethanol was added to 100 g of the dried product of black ganoderma and extracted at room temperature for 7 days, followed by filtration. The filtrate was concentrated to dryness to obtain 1.4 g of ganoderma ethanol extract.

Production Example 5 Butanol-soluble fraction of Ganoderma hot water extract 600 g of butanol was added to 6 g of Ganoderma hot water extract described in Production Example 1, extracted at room temperature for 7 days, filtered, and the filtrate was concentrated. Then, it was freeze-dried to obtain 0.6 g of a butanol-soluble fraction of the Ganoderma hot water extract (hereinafter referred to as a ganoderma hot water-butanol-soluble fraction).

Production Example 6 Ethyl acetate soluble fraction of Ganoderma hot water extract To 6 g of Ganoderma hot water extract described in Production Example 1, 600 mL of ethyl acetate was added, extracted at room temperature for 7 days, filtered, and the filtrate was filtered. Concentrated and freeze-dried to obtain 0.5 g of an ethyl acetate soluble fraction of the Ganoderma hot water extract (hereinafter referred to as a ganoderma hot water-ethyl acetate soluble fraction).

Comparative Example 1 Butanol-insoluble fraction of Ganoderma hot water extract The residue of the filtration step of Production Example 5 was dissolved in purified water, concentrated and freeze-dried, 5.0 g of a soluble fraction (hereinafter referred to as Ganoderma hydrothermal water-butanol insoluble fraction) was obtained.

Examples will be given to describe the present invention in detail, but the present invention is not limited thereto. In the examples,% indicates% by weight.

Soft capsule <Prescription>
Ingredient Amount (%)
1. Reishi hot water extract (Production Example 1) 30.0
2. Rice germ oil 60.0
3. Beeswah 7.0
4). Vitamin E 3.0
<Manufacturing method>
Ingredients 1 to 4 are mixed, and 250 mg is filled into a film composed of gelatin, glycerin, and caramel pigment, and after drying, a soft capsule is obtained.
<Usage>
Ingest 4 tablets per day.

Hard capsule <Prescription>
Ingredient Amount (%)
1. Ganoderma ethanol extract (Production Example 3) 45.0
2. Cornstarch 52.0
3. Sucrose fatty acid ester 3.0
<Manufacturing method>
Ingredients 1 to 3 are mixed and 250 mg is filled into a No. 2 hard capsule to obtain a capsule.
<Usage>
Ingest 4 tablets per day.

Beverage <Prescription>
Ingredient Amount (%)
1. Reishi hot water extract (Production Example 1) 1.0
2. Citric acid 6.0
3. Sucrose 10.0
4). Fragrance 1.0
5. Water 82.0
<Manufacturing method>
Components 1 to 4 are stirred and dissolved in a part of the component 5 water. The remaining water of component 5 is then added and mixed.
<Usage>
Take 50 mL per day.

Granule <Prescription>
Ingredient Amount (%)
1. Reishi hot water-butanol soluble fraction (Production Example 5) 15.0
2. Lactose 80.0
3. Cellulose 5.0
<Manufacturing method>
Ingredients 1 to 3 are granulated by a wet method and dried to obtain granules.
<Usage>
Take 1g per day.

Tablet <Prescription>
Ingredient Amount (%)
1. Reishi hot water-ethyl acetate soluble fraction (Production Example 6) 11.0
2. Lactose 60.0
3. Reduced maltose starch syrup 25.0
4). Sucrose fatty acid ester 4.0
<Manufacturing method>
Ingredients 1 to 4 are mixed and tableted to obtain 0.5 g of a tablet.
<Usage>
Take 2 capsules per day.

Cookies <Prescription> Blending amount (%)
1. Reishi ethanol extract (Production Example 3) 3.0
2. Rape oil 25.0
3. Honey 22.0
4). Soft flour 35.0
5. Whole grain 15.0
<Manufacturing method>
Ingredients 1-5 are mixed, molded, and baked at 170 degrees for 20 minutes.
<Usage>
Take 2 (20 g) daily.

Experimental Example 1- (1) Cholesterol Absorption Suppressing Effect in Rats with High Cholesterol Loading Male Wistar rats aged 3 weeks were divided into 5 groups so that their body weights were equal. A high cholesterol diet (hereinafter referred to as HC diet) was prepared by mixing powdered diet with 1% cholesterol and 0.25% sodium cholate, and the control group was fed with HC diet. For sample administration, HC feed was prepared by mixing 4% of Ganoderma hot water extract, Ganoderma ethanol extract, Ganoderma hot water-butanol soluble fraction, Ganoderma hot water-butanol insoluble fraction. It was done by taking it freely. Four days after the start of breeding, blood was collected from rats fasted overnight, and the total cholesterol level (hereinafter referred to as TC) was measured using plasma obtained by centrifugation, using a commercially available cholesterol measurement kit. The measurement results are shown as mean ± standard error, and Tukey's multiple comparison test was used for the significant difference test.

The results for total cholesterol are shown in Table 1. Cholesterol hot water extract, Ganoderma ethanol extract, and Ganoderma hot water-butanol soluble fraction were confirmed to have cholesterol absorption inhibitory action. On the other hand, it was not observed in the Ganoderma hydrothermal water-butanol insoluble fraction. When the Ganoderma hot water extract and the Ganoderma ethanol extract were compared, a statistically significant difference was observed, and the Ganoderma ethanol extract showed a remarkably strong cholesterol absorption inhibitory effect. In addition, when the Ganoderma hydrothermal extract was compared with the Ganoderma hydrothermal water-butanol soluble fraction, a statistically significant difference was observed, and the Ganoderma hydrothermal water-butanol soluble fraction was significantly stronger in suppressing cholesterol absorption. The effect was recognized.

Experimental Example 1- (2) LDL / HDL ratio lowering action in rats with high cholesterol loading In the rats described in Experimental Example 1- (1), HDL and triglyceride (hereinafter referred to as “HDL”) were obtained using plasma obtained 4 days after the start of breeding. The (TG) value was measured using a commercially available measurement kit. LDL calculated | required the value using the calculation formula (LDL = TC-HDL-0.2 * TG). The LDL / HDL ratio was calculated from the obtained LDL and HDL values. The results are shown as mean values ± standard error, and Tukey's multiple comparison test was used for the significance test.

The results of HDL, LDL value and LDL / HDL ratio are shown in Table 2. Reducing effect of LDL / HDL ratio was observed in Ganoderma hot water extract, Ganoderma ethanol extract, and Ganoderma hot water-butanol soluble fraction. On the other hand, it was not observed in the Ganoderma hydrothermal water-butanol insoluble fraction. When the Ganoderma hydrothermal extract and the Ganoderma hydrothermal water-butanol soluble fraction were compared, a statistically significant difference was observed, indicating that the Ganoderma hydrothermal water-butanol soluble fraction had a significantly strong LDL / HDL ratio. A lowering effect was observed.

Experimental Example 1- (3) Cholesterol excretion promoting action in rats loaded with high cholesterol In the rat described in Experimental Example 1- (1), feces were collected for 24 hours from the start of breeding to 2 to 3 days later and dried. Lipids were extracted from the obtained stool with chloroform / methanol (2: 1), and cholesterol in the stool was quantified by gas chromatography (GC-MS). The results are shown as mean values ± standard error, and Tukey's multiple comparison test was used for the significance test.

The results of the cholesterol excretion promoting action are shown in Table 3. Cholesterol excretion promoting action was observed in the Ganodermae ethanol extract and Ganoderma hot water-butanol soluble fraction. On the other hand, it was not observed in the Ganoderma hydrothermal water-butanol insoluble fraction. When the Ganoderma hot water extract and the Ganoderma ethanol extract were compared, a statistically significant difference was observed, and the Ganoderma ethanol extract showed a remarkably strong cholesterol excretion promoting effect. In addition, when the Ganoderma hydrothermal extract was compared with the Ganoderma hydrothermal water-butanol soluble fraction, a statistically significant difference was observed, and the Ganoderma hydrothermal water-butanol soluble fraction was significantly enhanced in promoting cholesterol excretion. The effect was recognized.

Experimental Example 2 Inhibition of Cholesterol Uptake into Lipid Micelle As final concentrations of 0.5 mM cholesterol, 0.6 mM phosphatidylcholine, 0.5 mM monoolein, and 1 mM oleic acid in chloroform / methanol (2: 1) solutions, respectively After mixing in a test tube, nitrogen gas was blown to distill off the solvent. 132 mM NaCl and 6.6 mM sodium taurocholate were dissolved in 15 mM phosphate buffer (pH 7.4) and added to the test tube. After sonication, the mixture was incubated at 37 ° C. for 24 hours to prepare a lipid micelle solution. Dispense this lipid micelle solution into another test tube, and use Ganoderma hot water extract, Ganoderma ethanol extract, Ganoderma hot water-butanol soluble fraction, Ganoderma hot water-butanol insoluble as samples. Fractions were added at 10 mg / mL and sonicated, and then incubated at 37 ° C. for 1 hour. After completion of the reaction, ultracentrifugation was performed to precipitate cholesterol that was not incorporated into the lipid micelles. The supernatant after centrifugation was collected and the amount of cholesterol was quantified by GC-MS. The percentage of cholesterol in the supernatant due to the addition of the sample when the amount of cholesterol in the control to which no reishi extract was added was taken as 100 was determined. The results are shown as mean values ± standard error, and Tukey's multiple comparison test was used for the significance test.

Table 4 shows the results of the action of suppressing the incorporation of cholesterol into lipid micelles. It means that the lower the proportion of cholesterol in the supernatant, the lower the cholesterol uptake into lipid micelles. Compared to the control (no sample added), the action of suppressing the uptake of cholesterol into lipid micelles was observed in the Ganoderma hot water extract, Ganoderma ethanol extract, and Ganoderma hot water-butanol soluble fraction. On the other hand, it was not observed in the Ganoderma hydrothermal water-butanol insoluble fraction. When the Ganoderma hydrothermal extract and the Ganoderma ethanol extract were compared, a statistically significant difference was observed, and the Ganoderma ethanol extract showed a remarkably strong cholesterol-inhibiting action on lipid micelles. In addition, when the Ganoderma hydrothermal extract was compared with the Ganoderma hydrothermal water-butanol soluble fraction, a statistically significant difference was observed, and the cholesterol lipid was significantly stronger in the Ganoderma hydrothermal water-butanol soluble fraction. Inhibition of uptake into micelles was observed.

The same test was conducted on the fraction dissolved in Ganoderma hydrothermal water-ethyl acetate (Production Example 6). As in the case of Ganoderma extract, cholesterol absorption inhibitory activity, LDL / HDL ratio lowering activity, cholesterol excretion Significant effects were observed on the promoting action and the action of suppressing the uptake of cholesterol into lipid micelles.

Using the drink of Example 3, 23 healthy men and women (40 to 60 years old) were subjected to a one month ingestion test, and 50 mL per day was ingested at an arbitrary time. Blood was collected before drinking and one month later, and a blood test was performed. As a result, a blood cholesterol level lowering effect and an LDL / HDL ratio lowering effect were observed. The blood test items indicating liver function and the like are within the normal range for all subjects, and it was confirmed that they have high safety.

From the above results, it was shown that the ganoderma extract has a cholesterol absorption inhibitory effect, a preventive or ameliorating effect of arteriosclerosis, a cholesterol excretion promoting action, and a cholesterol uptake inhibitory action. In addition, the effect is remarkably enhanced by changing the extraction solvent from water to a lower alcohol such as ethanol, and the effect is remarkably enhanced by the organic solvent-soluble fraction obtained by extracting Ganoderma hot water extract with an organic solvent. It has been shown.

The cholesterol absorption inhibitor of the present invention showed an excellent cholesterol absorption inhibitory action. Therefore, it is useful for use as a pharmaceutical, quasi-drug or food having a safe and excellent cholesterol absorption inhibitory effect.

Claims (8)

A cholesterol absorption inhibitor characterized by containing a ganoderma extract. A preventive or ameliorating agent for arteriosclerosis, comprising a reishi extract. Cholesterol excretion promoter characterized by containing reishi extract. An inhibitor of cholesterol uptake into lipid micelles, which contains a ganoderma extract. A cholesterol absorption inhibitor characterized by containing an organic solvent-soluble fraction of Ganoderma hydrothermal extract. A preventive or ameliorating agent for arteriosclerosis, comprising an organic solvent-soluble fraction of Ganoderma hot water extract. A cholesterol excretion enhancer comprising an organic solvent-soluble fraction of Ganoderma hydrothermal extract. An inhibitor of uptake of cholesterol into lipid micelles, comprising an organic solvent-soluble fraction of Ganoderma hydrothermal extract.