JP6460402B2 - Pharmaceutical composition or food composition for promoting cholesterol excretion - Google Patents

Description

  The present invention relates to a gramineous plant extract that promotes the in vitro elimination of cholesterol in foods and can suppress the absorption of the cholesterol in the body.

  In recent years, with the increase in energy intake due to Westernization of foods, an increase in lifestyle-related diseases such as obesity, fatty liver, hyperlipidemia, cardiovascular disease due to chronic excessive energy has become a social problem. Blood cholesterol levels are used as one of the diagnostic criteria for lifestyle-related diseases. High cholesterol is known as a risk factor for lifestyle-related diseases, particularly hyperlipidemia and cardiovascular diseases. The cause of lipid abnormalities such as hypercholesterolemia is due to genetic factors, but is often caused by lifestyle habits such as eating habits. From the viewpoint of improving the dietary habits that cause it and simplicity, it is desirable if cholesterol can be lowered through daily diet.

  Conventionally, the cholesterol lowering effect by dietary fiber is known. This effect is attributed to the fact that cholesterol is lowered by promoting the synthesis of bile acids from cholesterol in the body by reducing the concentration of bile acids in the digestive tract by dietary fiber. However, since dietary fiber is bulky and has a poor taste, it is difficult to continuously take the necessary amount in daily meals.

  Regarding a material having a cholesterol lowering action, Patent Document 1 discloses that an insoluble component obtained from a proteolytic enzyme degradation product of gluten has an excellent ability to adsorb bile acids, promotes bile acid excretion and lowers cholesterol. It describes that it has an action, and also describes a cholesterol-lowering composition containing the insoluble component, and a medicine, food and drink, and feed containing the composition. Patent Document 2 discloses a liquid composition obtained by removing phytohaemagglutinin by heat treatment of an aqueous extract of wheat flour or wheat bran, and further removing a low molecular weight substance having a molecular weight of 10,000 or less, or evaporating to dryness. It is described that the powder obtained in this way has a neutral fat lowering action and a total cholesterol lowering action. The inventions described in Patent Documents 1 and 2 are basically considered to contain a relatively large amount of protein, and are hardly called low proteins.

JP 2012-140396 A Japanese Patent Laid-Open No. 01-312975

  The cholesterol lowering effect of conventional materials has not always been sufficiently satisfactory. No material has yet been provided that is excellent in the effect of suppressing the absorption of cholesterol in foods, is safe and has few side effects, and can be used as a medicine, food and drink, feed and the like.

  The present invention relates to a gramineous plant extract having a cholesterol excretion promoting ability that is safe and has few side effects.

  As a result of intensive studies to solve the above problems, the present inventors have found that an ethanol extract of wheat bran, which is a kind of gramineous plant, containing alkylresorcinol has an ability to promote cholesterol extracorporeal excretion (bile acid adsorption). It was found that cholesterol in the food is promoted to be discharged out of the body together with the stool by ingesting it together with the food.

This invention is made | formed based on the said knowledge, and is a Gramineae plant extract which has the ability to promote cholesterol excretion, extracted from a Gramineae plant using alcohol.
Moreover, this invention is the pharmaceutical or food-drinks containing the said Gramineae plant extract.

  The gramineous plant extract of the present invention has an ability to promote cholesterol excretion, and promotes the excretion of cholesterol in foods together with feces, thereby effectively suppressing the absorption of cholesterol in the body. In addition, the gramineous plant extract of the present invention is mainly composed of ingredients contained in gramineous plants such as wheat and rye that have abundant dietary experience. There are few concerns that cause harmful effects on the living body, and it is highly safe and has a low risk of side effects.

FIG. 1 is a graph showing the amount of feces per body weight of each mouse group in a cholesterol excretion promoting action confirmation test. FIG. 2 is a graph showing the amount of cholesterol in the stool of each group of mice in the cholesterol excretion promoting action confirmation test. FIG. 3 is a graph showing the amount of free bile acid in a sample in a bile acid adsorption capacity confirmation test, and the smaller the amount, the bile acid of the test subject (the grass plant extract of the example, phosphatidylcholine). It shows that adsorption capacity is high. FIG. 4 is a graph showing the amount of free cholesterol in a sample in a bile acid-cholesterol micelle formation inhibitory confirmation test, and the larger the amount, the more the test object (the grass plant extract of the example, phosphatidylcholine). It shows that the ability to inhibit micelle formation between bile acid and cholesterol is high.

  The gramineous plant extract of the present invention having the ability to promote cholesterol excretion is an alcoholic extract of gramineous plants. Grains such as wheat, rye, rye, barley, oats, corn, corn, barnyard millet, millet, millet, and the like can be cited as examples of Gramineae plants that are extraction sources. Species can be used alone or in combination of two or more. Among these cereals, wheat and rye are particularly preferable. The type of wheat is not particularly limited. Moreover, the gramineous plant which is an extraction source normally uses the fruit, and when the gramineous plant which is an extraction source is wheat, wheat grain is used. In the present invention, berries of gramineous plants or dried products thereof may be used as an extraction source while maintaining the shape of the grains, or strips obtained by cutting, pulverizing, etc. A pulverized product (powder) may be used as an extraction source.

  It is preferable that the Gramineae plant which is an extraction source contains an outer skin (fruit skin, seed coat) of fruit juice. Wheat bran is preferable from the standpoints of availability, price, and the like as wheat flour, rye whole flour, wheat bran, and rye hulls that are preferably used as the extraction source in the present invention.

  Examples of alcohols used for the extraction of gramineous plants include monovalent lower alcohols (preferably those having 1 to 4 carbon atoms) such as methanol, ethanol, n-propanol, isopropanol, n-butanol, and 1,3. -Alcohol which is liquid at room temperature (25 degreeC), such as polyhydric alcohols, such as butylene glycol, propylene glycol, and glycerol. As used herein, the alcohol includes water-containing alcohols containing aqueous components such as water, pure water, distilled water, tap water, acidic water, alkaline water, and neutral water in addition to alcohol. The alcohol content in the hydrous alcohol is usually 70% by volume or more, preferably 80% by volume or more, more preferably 90% by volume or more. Among these alcohols, ethanol (hydrous ethanol) is particularly preferable from the viewpoint of operability and environmental performance.

  The method for extracting grass plants with alcohol is not particularly limited. For example, a method of immersing, stirring, or refluxing fruit fruits such as wheat grains or a fragment or pulverized product thereof in alcohol, supercritical fluid extraction method, etc. Can be mentioned. For example, in the case of a method in which wheat bran is immersed, stirred or refluxed in ethanol (hydrous ethanol), the extraction temperature (ethanol liquid temperature) is preferably 2 to 80 ° C., the extraction time is 0.5 to 72 hours, and ethanol is used. The amount is preferably 50 to 2000 parts by mass with respect to 100 parts by mass of wheat bran.

  The grass plant extract of the present invention preferably contains alkylresorcinol as an active ingredient. According to the knowledge of the present inventors, the cholesterol excretion promoting ability of the grass plant extract of the present invention is largely due to the action of alkylresorcinol represented by the following general formula (I). The grass plant extract of the present invention preferably contains one or more alkylresorcinol represented by the following general formula (I) as an active ingredient. The content of alkylresorcinol represented by the following general formula (I) in the grass plant extract of the present invention is the grass plant extract of the present invention from the viewpoint of more surely exerting a cholesterol excretion promoting effect. Among them, the content is preferably 70% by mass or more, more preferably 80% by mass or more. The content of the alkylresorcinol represented by the following general formula (I) is 100% by mass, that is, the grass plant extract of the present invention may be composed of only the alkylresorcinol represented by the following general formula (I). good.

Regarding R ₁ in the general formula (I), examples of the saturated alkyl group having 15 to 25 carbon atoms include n-pentadecyl, n-heptadecyl, n-nonadecyl, n-henicosyl, n-tricosyl, n- Examples include linear ones such as pentacosyl and n-heptacosyl, and besides these, branched or cyclic ones may be used. Among these, a saturated alkyl group having 15 to 23 carbon atoms is preferable.

Moreover, regarding R ₁ in the general formula (I), examples of the unsaturated alkyl group having 15 to 25 carbon atoms include those corresponding to the saturated alkyl group having 15 to 25 carbon atoms. There is no restriction | limiting in particular in the number and position of the unsaturated bond contained in an unsaturated alkyl group.

In the general formula (I), R ₂ is preferably a hydrogen atom, and R ₁ is preferably bonded to R ₂ at the para position.

Specific examples of the alkyl resorcinol that can be contained in the grass plant extract of the present invention include the following.
1,3-dihydroxy-5-n-pentadecylbenzene (C15: 0)
1,3-dihydroxy-5-n-heptadecylbenzene (C17: 0)
1,3-dihydroxy-5-n-nonadecylbenzene (C19: 0)
1,3-dihydroxy-5-n-henicosylbenzene (C21: 0)
1,3-dihydroxy-5-n-tricosylbenzene (C23: 0)
1,3-dihydroxy-5-n-pentacosylbenzene (C25: 0)

As a preferable example of the gramineous plant extract of the present invention, those containing the following 6 types of alkylresorcinol can be mentioned.
1) Alkyl resorcinol (hereinafter, also referred to as AR15) in which R ₁ in the general formula (I) is a saturated or unsaturated alkyl group having 15 carbon atoms.
2) Alkyl resorcinol (hereinafter, also referred to as AR17) in which R ₁ in the general formula (I) is a saturated or unsaturated alkyl group having 17 carbon atoms.
3) Alkyl resorcinol (hereinafter, also referred to as AR19) in which R ₁ in the general formula (I) is a saturated or unsaturated alkyl group having 19 carbon atoms.
4) Alkyl resorcinol (hereinafter, also referred to as AR21) in which R ₁ in the general formula (I) is a saturated or unsaturated alkyl group having 21 carbon atoms.
5) Alkyl resorcinol (hereinafter also referred to as AR23) in which R ₁ in the general formula (I) is a saturated or unsaturated alkyl group having 23 carbon atoms.
6) Alkyl resorcinol (hereinafter also referred to as AR25) in which R ₁ in the general formula (I) is a saturated or unsaturated alkyl group having 25 carbon atoms.

Particularly preferred as AR15 is one in which R ₁ is a saturated alkyl group having 15 carbon atoms and R ₂ is a hydrogen atom. Specifically, 1,3-dihydroxy-5-n-pentadecylbenzene (C15 : 0).
Particularly preferred as AR17 is one in which R ₁ is a saturated alkyl group having 17 carbon atoms and R ₂ is a hydrogen atom. Specifically, 1,3-dihydroxy-5-n-heptadecylbenzene (C17 : 0).
Particularly preferred as AR19 is one in which R ₁ is a saturated alkyl group having 19 carbon atoms and R ₂ is a hydrogen atom. Specifically, 1,3-dihydroxy-5-n-nonadecylbenzene (C19 : 0).
Particularly preferred as AR21 is one in which R ₁ is a saturated alkyl group having 21 carbon atoms and R ₂ is a hydrogen atom. Specifically, 1,3-dihydroxy-5-n-henecosylbenzene ( C21: 0).
Particularly preferred as AR23 is one in which R ₁ is a saturated alkyl group having 23 carbon atoms and R ₂ is a hydrogen atom. Specifically, 1,3-dihydroxy-5-n-tricosylbenzene (C23 : 0).
Particularly preferred as AR25 is one in which R ₁ is a saturated alkyl group having 25 carbon atoms and R ₂ is a hydrogen atom. Specifically, 1,3-dihydroxy-5-n-pentacosylbenzene (C25 : 0).

In the gramineous plant extract of the present invention, the contents of AR15, AR17, AR19, AR21, AR23, and AR25 are each preferably within the following ranges from the viewpoint of further improving the ability to promote cholesterol excretion.
The content of AR15 is preferably 0.1 to 10.0% by mass, more preferably 0.1 to 5.0% by mass, and particularly preferably 0.5 to 1.% by mass in the grass plant extract of the present invention. 5% by mass.
The content of AR17 is preferably 1.0 to 20.0% by mass, more preferably 5.0 to 15.0% by mass, and particularly preferably 8.0 to 12% in the grass plant extract of the present invention. 0% by mass.
The content of AR19 is preferably 25.0 to 40.0% by mass, more preferably 27.5 to 37.5% by mass, and particularly preferably 30.0 to 35.% in the grass plant extract of the present invention. 0% by mass.
The content of AR21 is preferably 40.0 to 55.0% by mass, more preferably 42.5 to 52.5% by mass, and particularly preferably 45.0 to 50.% in the grass plant extract of the present invention. 0% by mass.
The content of AR23 is preferably 1.0 to 15.0% by mass, more preferably 2.5 to 12.5% by mass, and particularly preferably 5.0 to 10% by mass in the grass plant extract of the present invention. 0% by mass.
The content of AR25 is preferably 0 to 5.0% by mass, more preferably 0 to 2.0% by mass, and particularly preferably 0 to 1.5% by mass in the grass plant extract of the present invention.

The grass plant extract of the present invention may contain one or more alkylresorcinols other than AR15, AR17, AR19, AR21, AR23, and AR25. Examples of the other alkylresorcinol include alkylresorcinol (hereinafter, also referred to as AR27) in which R ₁ in the general formula (I) is a saturated or unsaturated alkyl group having 27 carbon atoms. Particularly preferred as AR27 is one in which R ₁ is a saturated alkyl group having 27 carbon atoms and R ₂ is a hydrogen atom. Specifically, 1,3-dihydroxy-5-n-heptacosylbenzene ( C27: 0).

  The grass plant extract of the present invention preferably has a high content of alkylresorcinol having the ability to promote cholesterol excretion. From this viewpoint, the extract extracted from the grass plant using alcohol is distributed chromatography. It is preferable to purify by, for example, or concentrate and evaporate to dryness by a conventional method. Examples of partition chromatography used to purify the alcoholic extract of Gramineae include normal phase chromatography using a non-aqueous solvent as the mobile phase, such as open column method, medium pressure column method, high performance liquid chromatography, etc. These known methods can be appropriately selected.

  As a mobile phase in partition chromatography of an alcoholic extract of a grass family, monovalent lower alcohols (preferably having 1 to 4 carbon atoms) such as methanol, ethanol, n-propanol, isopropanol, n-butanol, And alcohols that are liquid at room temperature (25 ° C.) such as polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, and glycerin; ethers such as diethyl ether and propyl ether; esters such as butyl acetate and ethyl acetate; Examples include ketones such as ethyl methyl ketone; hexane; methylene chloride; acetonitrile; and chloroform. One of these solvents can be used alone, or two or more can be used in combination. When a plurality of solvents are combined to form a mobile phase, an isocratic mode in which the mixing ratio of the plurality of solvents is kept constant during partition chromatography (purification of the gramineous alcohol extract), or A gradient mode in which the mixing ratio is changed may be used. Any carrier can be used as the carrier in the partition chromatography as long as it can carry and release the target active ingredient, and generally includes silica gel, polyacrylamide gel, dextran gel and the like. . The detection wavelength in the partition chromatography of the alcoholic extract of the gramineous plant should just be 170-320 nm, Preferably it is 190-280 nm.

  The Gramineae plant extract of the present invention does not contain a protein or has a relatively low content even if it contains a protein, and has a low protein content. In particular, when the Gramineae plant that is the source of extraction is even wheat bran, the Gramineae plant extract extracted therefrom has a low protein content with a very low protein content. The content of the protein in the gramineous plant extract of the present invention depends on the kind of gramineous plant as the extraction source, but is preferably 5% by mass or less, more preferably in the gramineous plant extract of the present invention. 2% by mass or less. Since the gramineous plant extract of the present invention is low in protein, it is also suitable for patients with kidney diseases and the like who need to limit the intake of protein.

  The grass plant extract of the present invention can be used as a pharmaceutical product, food or drink, feed, etc. for humans or animals, or for producing them, taking advantage of its ability to promote cholesterol excretion. The form of the grass plant extract of the present invention is not particularly limited. For example, a liquid material (liquid or semi-liquid) containing a grass plant alcohol extract (alkyl resorcinol represented by the general formula (I)), Examples include a dry solid obtained by evaporating and drying a liquid medium (alcohol or hydrous alcohol) in the liquid or a powder thereof, an alcohol solution obtained by dissolving the dry solid or the powder in an alcohol such as ethanol, and the like. It is done. The present invention also provides a pharmaceutical, a food and drink, and a feed containing the grass plant extract of the present invention.

  The pharmaceutical product of the present invention, that is, the cholesterol excretion promoter, can be a serum cholesterol lowering agent or a therapeutic agent for hypercholesterolemia containing the grass plant extract of the present invention as an active ingredient. Moreover, the pharmaceutical of this invention can be administered by arbitrary dosage forms including oral and parenteral administration. Dosage forms for oral administration include solid preparations such as tablets, coated tablets, granules, powders, capsules, or liquid preparations such as elixirol, syrups and suspensions. Examples of the dosage form for parenteral administration include injection, infusion, transdermal, transmucosal, nasal, enteral, inhalation, suppository, bolus and the like.

  The food / beverage product or feed of the present invention contains the grass plant extract of the present invention as an active ingredient, and is intended to have an effect of promoting the in vitro elimination of cholesterol (inhibition effect of absorption in the body), and the function indicating that effect. It may be a food or drink, a food or drink for the sick, a food or drink for specified health use, a livestock feed, a pet food or the like. The form of the food or drink or feed of the present invention is not particularly limited, and examples of the form of the beverage include tea drinks, coffee drinks, milk drinks, fruit drinks, carbonated drinks, alcoholic drinks, soft drinks, and the like. Alternatively, the form of the feed can be solid, semi-solid or liquid, and includes tablet form, pill form, capsule form, liquid form, syrup form, powder form, granule form and the like. Specific food forms include breads, noodles, jelly-like foods and various snacks, baked goods, cakes, chocolate, gum, candy, tablets, capsules, soups, dairy products, frozen foods, instant foods, Examples include supplements, other processed foods, seasonings, and materials thereof.

The pharmaceutical, food and drink, or feed of the present invention may contain other ingredients other than the grass plant extract of the present invention (alkylresorcinol represented by the general formula (I)), for example, other effective, as necessary. It may contain components and / or commonly used additives or carriers.
In the case of pharmaceuticals, examples of the other ingredients include other medicinal ingredients, cosmetic ingredients, nutrition ingredients, and excipients, disintegrants, binders, lubricants, surfactants, water-soluble polymers, and diluents. , Osmotic pressure adjusting agent, pH adjusting agent, emulsifier, preservative, buffer, stabilizer, antioxidant, thickener, ultraviolet absorber, activity enhancer, colorant, sweetener, flavoring agent, flavoring agent, A sour agent etc. are mentioned, These 1 type can be used individually or in combination of 2 or more types.
In the case of food and drink or feed, the other ingredients include, for example, other food and drink materials, nutritional ingredients, cosmetic ingredients, other functional materials and solvents, oils, softeners, emulsifiers, preservatives, stabilizers, oxidation Examples thereof include additives such as an inhibitor, a colorant, a humectant, a thickener, a brightener, a sweetener, and a fragrance, and these can be used alone or in combination of two or more.

  The intake of the grass plant extract of the present invention (alkylresorcinol represented by the general formula (I)) in the pharmaceutical, food and drink of the present invention varies depending on their dosage form and form, In any case of food and drink and feed, it is preferably ingested so as to be 0.01 to 10 g per day.

The present invention includes the following <1> to <3> modes.
<1> A food or beverage or feed additive having an ability to promote in vitro elimination of cholesterol, which is used by adding to a food or beverage or feed, including a grass plant extract extracted from a grass plant using alcohol.
<2> A method for promoting in vitro elimination of cholesterol by administering to a living body (mammal) a grass plant extract extracted from a grass plant using alcohol.
<3> By discharging a gramineous plant extract extracted from a gramineous plant with alcohol into food and ingesting it into a living body (mammal), the cholesterol in the food is discharged out of the body together with feces. How to promote.

  EXAMPLES Hereinafter, although an Example and a test example are given and this invention is demonstrated further in detail, this invention is not restrict | limited by these Examples and a test example.

〔Example〕
By the following <Extraction and purification method>, a Gramineae plant extract (wheat extract) containing an alkylresorcinol represented by the general formula (I) was obtained. The composition of this grass plant extract is as follows.
-1,3-dihydroxy-5-n-pentadecylbenzene (C15: 0) 1.2 mass%.
-1,3-dihydroxy-5-n-heptadecylbenzene (C17: 0) 10.9 mass%.
-1,3-dihydroxy-5-n-nonadecylbenzene (C19: 0) 33.9 mass%.
-1,3-dihydroxy-5-n-henicosylbenzene (C21: 0) 46.4 mass%.
-7.5 mass% of 1, 3- dihydroxy-5-n-tricosyl benzene (C23: 0).
-1,3-dihydroxy-5-n-pentacosylbenzene (C25: 0) 0.1 mass%.

<Extraction purification method>
Ethanol in an amount of 5 times by mass was added to wheat bran, and the mixture was extracted by stirring for 16 hours under conditions of 600 rpm and room temperature. The extract was filtered to remove unnecessary substances and an ethanol extract was collected, and then ethanol was distilled off to obtain an ethanol extract of wheat bran. The ethanol extract was then purified by medium pressure chromatography. Medium pressure chromatography conditions are as follows. A peak component appearing 31 to 36 minutes after the start of elution was collected and the solvent was distilled off to obtain a grass plant extract containing the alkylresorcinol represented by the general formula (I).
(Medium pressure chromatography conditions)
Column: silica gel (injection column 3 L, high flash column 5 L, 60 mm, 40 μm, manufactured by Yamazen Corporation)
・ Mobile phase: Hexane / ethyl acetate mixed solvent (volume ratio) = 9/10 at 90/10, 15 minutes at 80/20, 16 minutes at 60/40 ・ Detection wavelength: 254 nm

In addition, the purification of the wheat bran ethanol extract in the <Extraction and purification method> can be performed by HPLC instead of medium pressure chromatography. In that case, methanol is added to the ethanol extract to prepare a methanol additive solution having a concentration of the ethanol extract of 200 μg / ml, and the methanol additive solution is passed through a filter having a pore size of 0.45 μm. , HPLC sample. HPLC conditions are as follows.
(HPLC conditions)
Column: silica gel (ODS-80A, 5 μm, 4.6 × 250 mm, manufactured by GL Sciences Inc.)
Guard column: ODS-80A, 5 μm, 4.6 × 50 mm,
-Column temperature: 30 ° C
-Mobile phase: 100% methanol
・ Detection wavelength: 215 nm

[Test Example 1]
About the gramineous plant extract of an Example, the influence on the cholesterol of a high fat high sucrose diet load mouse | mouth was investigated by the following test. The results are shown in FIGS. 1 and 2, ND is a group of mice fed a normal diet, HFHSD is a group of mice fed a high-fat high sucrose diet, and HFHSDAR is added with the grass plant extract of the example. A group of mice fed a high fat, high sucrose diet.

<Cholesterol excretion promoting action confirmation test>
As a normal diet (ND), AIN93M (using milk casein) (made by Oriental Yeast Co., Ltd.) and F2HFHSD (made by Oriental Yeast Industry Co., Ltd.) as a high fat high sucrose diet (HFHSD) are prepared. Prepared by adding 0.5% by mass of the gramineous plant extract of Example (HFHSDAR) was prepared. C57BL / 6JJmsSlc strain mice (4-week-old male, Japan SLC Co., Ltd.) were bred for 2 weeks under a light-dark cycle of 12 hours light and 12 hours dark (lights on at 0:00, lights off at 12:00). Later, these mice were divided into ND feeding group (2 gauges each with 6 gauges), HFHSD feeding group (2 gauges each with 6 gauges), and HFHSDAR feeding group (2 gauges with 6 gauges each). And were allowed to eat freely for 10 weeks. After the free feeding period, all feces for one day in each gauge were collected, and the collected feces were dried at 40 ° C. for 16 hours with a dryer to obtain dried feces. 0.2 ml of 50 μmol / L NaCl was prepared, and 0.1 g of dried feces was added and suspended therein, 2 ml of ethanol was added to the suspension, and the dried feces were pulverized with a homogenizer. The pulverization of the dried stool was performed by vigorously shaking the suspension while heating the suspension containing the dried stool to 80 ° C. After pulverizing the dried feces, the suspension was centrifuged at 5800 × g for 15 minutes, and the supernatant was collected and evaporated, and 0.2 ml of isopropanol containing 20% by mass of Triton X-100 was added. A solution of the residue was obtained, and the amount of cholesterol in the solution was measured using Kit (Wako Pure Chemical Industries).

FIG. 1 is a graph showing the amount of stool (average value) per body weight of each group of mice in a cholesterol excretion promoting action confirmation test. In FIG. 1, ND (ordinary diet group) has an average value of 0.00771. The standard error is 0.00037, the HFHSD (high fat high sucrose diet group) has an average value of 0.00848, the standard error is 0.00035, and HFHSDAR (the gramineous plant extract of Example is added) The high fat high sucrose diet group) had an average value of 0.00848 and a standard error of 0.00039.
FIG. 2 is a graph showing the amount of stool cholesterol (average value) in each mouse group in the same test. In FIG. 2, ND has an average value of 12.937 and a standard error of 0.504, and HFHSD is The average value was 30.520, the standard error was 1.181, and the HFHSDAR had the average value of 42.067 and the standard error of 2.715.

As is clear from FIG. 1, there was no substantial difference in the amount of feces per body weight among the ND feeding group, the HFHSD feeding group, and the HFHSDAR feeding group. In the HFHSDAR feeding group, the amount of bile acids in stool was not increased.
Based on this, when looking at FIG. 2, the amount of cholesterol in the stool of the HFHSD feeding group is significantly increased compared to that of the ND feeding group, and therefore, continuous intake of HFHSD is a factor in the absorption of cholesterol in the body. Clearly it can be the cause. This is presumably because F2HFHSD used as HFHSD contains beef tallow and lard which are not included in AIN93M used as ND.
On the other hand, as is clear from FIG. 2, the amount of stool cholesterol in the HFHSDAR fed group was significantly increased compared to that in the HFHSD fed group. It is clear that the plant family extract has an action of promoting cholesterol excretion.

[Test Example 2]
About the Gramineae plant extract of an Example, bile acid adsorption ability was evaluated by the following test. The result is shown in FIG. In FIG. 3, ** (two asterisks) indicates that the significant difference between those without asterisks (addition amounts of Gramineae plant extracts 0 mg, 0.1 mg, 1 mg) is significant level p < It is recognized as 0.01.

<Bile acid adsorption capacity confirmation test>
Five types of mixed solutions containing PBS (pH 7.4) and bile acid (taurocholic acid) and the grass plant extract or phosphatidylcholine of Examples were prepared. The prepared mixed solutions are common at a PBS concentration of 0.1 M and a bile acid concentration of 2 mM, but the addition amount of the Gramineae plant extract is different, and the addition amount is 0 mg, 0.1 mg, 1 mg, There are five types of 10 mg and 20 mg. Phosphatidylcholine was added to a mixed solution containing 0 mg of Gramineae extract so that its concentration was 0.6 mM.
Each mixed solution was sonicated, incubated at 37 ° C. for 2 hours, and then centrifuged at room temperature (25 ° C.) at a rotational speed of 13500 rpm for 15 minutes. The centrifuged supernatant was collected, and a syringe filter (0 .22 μm) to separate the filtrate and the residue, and the concentration of bile acid in the filtrate (the concentration of free bile acids not bound to other components) was measured. Four samples were prepared for each liquid mixture, and the bile acid concentration was measured for each sample (n = 4).

  As is clear from FIG. 3, the system in which the amount of the grass plant extract of Example is 10 mg or more binds to other components as compared with the system in which the amount is 1 mg or less and the system in which phosphatidylcholine is added. The concentration of free bile acids not significantly decreased. This is because the presence of a certain amount or more of the Gramineae extract of the example with respect to bile acid promotes the formation of micelles by the combination of both, and the micelles precipitate in the mixed solution and are contained in the residue. , To be removed from the filtrate. From this result, it was suggested that the Gramineae plant extract of the Example has the ability to bind to bile acids (bile acid adsorption ability). In the Gramineae extract of the examples, it is considered that alkylresorcinol binds to bile acids.

[Test Example 3]
About the Gramineae plant extract of an Example, bile acid-cholesterol micelle formation inhibitory ability was evaluated by the following test. The result is shown in FIG. In addition, in FIG. 4, * (one asterisk) is a significant level p <0 significant difference between what is not attached with an asterisk (addition amount of Gramineae plant extract 0 mg, 0.1 mg, 1 mg). .05 (represented by two asterisks) indicates that a significant difference from that without the asterisk was recognized at the significance level p <0.01.

<Bile acid-cholesterol micelle formation inhibitory ability confirmation test>
Six types of mixed solutions containing PBS (pH 7.4) and bile acid (tauryl cholic acid) and the grass plant extract or phosphatidylcholine of Examples were prepared. The prepared mixed solutions are common at a PBS concentration of 15 mM and a bile acid concentration of 6.6 mM, but the addition amount of the gramineous plant extract is different. The addition amount is 0 mg, 0.1 mg, 1 mg, There are 6 types of 4 mg, 8 mg and 10 mg. Phosphatidylcholine was added to a mixed solution containing 0 mg of Gramineae extract so that its concentration was 2.4 mM.
Each mixed solution was sonicated, incubated at 37 ° C. for 16 hours, and then centrifuged at room temperature (25 ° C.) at a rotational speed of 13500 rpm for 15 minutes. The centrifuged supernatant was collected, and a syringe filter (0 .22 μm) to separate the filtrate and the residue, and the concentration of cholesterol in the filtrate (the concentration of free cholesterol not bound to other components) was measured. Four samples were prepared for each liquid mixture, and the cholesterol concentration was measured for each sample (n = 4).

  As is clear from FIG. 4, the system in which the amount of the grass plant extract of Example is 4 mg or more binds to other components as compared with the system in which the amount is 1 mg or less and the system in which phosphatidylcholine is added. Not free cholesterol concentration increased significantly. This indicates that when a certain amount or more of the Gramineae plant extract of Example is present with respect to bile acids, cholesterol that is not taken into micelles due to the combination of bile acids and cholesterol increases (bile acid-cholesterol). Micelles are precipitated in the mixture and are included in the residue and are removed from the filtrate). That is, when a certain amount or more of the Gramineae plant extract of the Example is present with respect to bile acids, the formation of micelles by the combination of bile acids and Gramineae plant extracts is promoted, while bile acids and cholesterol and It is considered that the formation of micelles due to the binding of is inhibited, and as a result, the in vitro elimination of cholesterol is promoted. In the gramineous plant extracts of Examples, it is considered that alkylresorcinol inhibits the formation of bile acid-cholesterol micelles.

Claims (4)

A pharmaceutical composition or food composition for promoting cholesterol excretion comprising an alcoholic extract of wheat bran as an active ingredient. The pharmaceutical composition or food composition for promoting cholesterol excretion according to claim 1, comprising alkylresorcinol. The pharmaceutical composition or food composition for promoting cholesterol excretion according to claim 1 or 2, comprising an alkylresorcinol represented by the following general formula (I) .
The content of protein in the alcohol extract is 5% by mass or less, The pharmaceutical composition or food composition for promoting cholesterol excretion according to any one of claims 1 to 3 .