JP3977889B2 - Drugs containing buckwheat husk extract as an active ingredient - Google Patents

Description

[0001]
[Industrial application fields]
The present invention relates to an extract obtained from buckwheat husk, a component having a molecular weight of 10,000 or less obtained by fractionating and purifying this, and a lipid peroxide inhibitor comprising a polyphenol fraction obtained by further fractionation thereof as an active ingredient, The present invention relates to a lipid peroxide inhibitor, a cholesterol-lowering agent, a neutral fat-lowering agent, and a hyperlipidemia-improving agent.
[0002]
[Prior art]
Soba is one of the most popular foods used in Japan for a long time. In addition, as familiar as New Year's soba and moving soba, Japanese people are highly health-conscious with respect to buckwheat. In fact, rutin is known as a component that strengthens capillaries and is effective for hypertension. In animal experiments, etc., effects are recognized for hypertension, arteriosclerosis, diabetes and the like (Journal of Women's Nutrition University, Vol. 22, 25-30 (1991), etc.). Buckwheat has been cultivated as a salvage crop since ancient times, and the endosperm portion obtained from seeds has been floured and edible as buckwheat flour. The seed coat (buckwheat husk) removed at this time is used as a filling material for pillows, and most of the seed coat is currently disposed of as incinerators. Moreover, almost no effective use is performed about the component contained in a buckwheat husk. It is known that buckwheat contains flavonoids such as rutin as well as high-quality protein, vitamin B group, dietary fiber. As for buckwheat husks, there are not many reports on ingredients, but it is considered that polyphenols are contained in addition to the same ingredients as buckwheat.
[0003]
Polyphenolic compounds are known to exist in the plant kingdom as a secondary metabolite of plants in a wide variety and abundantly, but these compounds have long been known for their diverse biological activities. In the field of phytochemistry and the like, and in recent years in the field of health food. In particular, tea polyphenols have been actively researched, and antibacterial, antiviral, antimutagenic, antioxidant, antihypertensive, antihypertensive, hypocholesterolemic, anticholesterolic, in tea polyphenols, especially catechins. It is known to have a very wide range of physiological activities such as caries, antiallergy, improvement of intestinal flora, and deodorization.
[0004]
Today, the number of patients with brain diseases such as adult diseases, cancer, allergies, and dementia continues to increase. In particular, it is pointed out that the number of patients with these diseases, especially dementia and Alzheimer syndrome, will increase in the future. ing. In relation to these, it has been pointed out that active oxygen species generated in vivo are involved. In addition, due to changes in dietary habits, a wide range of age groups from children to adults has recently been pointed out that excessive intake of fat has been pointed out, so that triglycerides and cholesterol levels in the blood exceed the normal range. Cholesterolemia is increasing. These symptoms have been pointed out to be related to adult diseases, and are also the cause of organ damage such as brain, heart and liver, hypertension and diabetes.
[0005]
In recent years, there has been an increasing interest in useful substances in foods, and health foods that have no side effects and can be consumed with peace of mind are becoming a boom. Particularly for elderly people, infants, children, etc., there is a desire for a method of treatment and prevention with health foods that have no side effects.
[0006]
For buckwheat husks, extract with different polar solvents (petroleum ether, ethyl acetate, ethanol), evaluate the antioxidation by measuring the lipid peroxidation rate when the extract is added to the lipid, and extract the ethanol extract The highest activity is observed, and a high negative correlation is observed between the total polyphenol content in the ethanol extract and the lipid peroxidation rate, and the lipid peroxidation rate of the buckwheat extract is 10 times that of α-tocopherol. Although there is a report that it is 1/20 to 1/20 (Journal of Japanese Society for Food Chemical Engineering, 42,649 (1995)), no examples of active use as pharmaceuticals or health foods have been found so far.
[0007]
[Problems to be solved by the invention]
Thus, it is necessary to find an active ingredient from a food or something equivalent to food in order to treat and prevent an aging society and a disease associated therewith that are expected in the future. Under such circumstances, an object of the present invention is to effectively utilize bioactive components in buckwheat husks that have not been used as food wastes, food additives, etc. until now.
[0008]
[Means for solving the problems]
The present inventors have intensively studied in view of the above-mentioned object, and have studied an efficient fractionation method of bioactive fractions contained in buckwheat husks and a method for producing bioactive polyphenols, and perform operations such as extraction and separation. It was confirmed by animal experiments that the extract obtained by the above has a physiological activity that has not been known so far, and the present invention has been completed.
[0009]
That is, the present invention relates to 1) a lipid peroxide inhibitor, a cholesterol-lowering agent, a neutral fat-lowering agent or an active ingredient containing an extract obtained by subjecting buckwheat husk to water or an organic solvent alone or a mixture thereof. Hyperlipidemia improving agent,
2) The lipid peroxide inhibitor, cholesterol lowering agent, neutral fat lowering agent or high as described in 1 above, wherein the component obtained by fractionating and purifying an extract obtained by extraction treatment of buckwheat husk is an active ingredient. Lipemia improving agent,
3) The lipid peroxide inhibitor, cholesterol-lowering agent, neutral fat-lowering agent or hyperlipidemia according to 1 above, wherein the polyphenol fraction obtained by fractionating and purifying the extract obtained by extracting the buckwheat husk is an active ingredient Improver,
4) The lipid peroxide inhibitor according to any one of 1 to 3 above,
5) The cholesterol-lowering agent according to any one of 1 to 3 above,
6) The neutral fat lowering agent according to any one of 1 to 3 above, and 7) the hyperlipidemia improving agent according to any one of 1 to 3 above.
[0010]
Hereinafter, the present invention will be described in detail.
The buckwheat husk extract in the present invention, the active fraction of low molecular weight (molecular weight of 10,000 or less) in the extract, and the polyphenol fraction obtained by fractionating and purifying the extract are water or organic An extract obtained by extraction with a solvent alone or a mixture thereof, and filtration, concentration, or the like, or an extract obtained by subjecting the extract to adsorption, separation, ultrafiltration, or the like.
[0011]
In the extraction, water, alcohol, a hydrophilic organic solvent, or the like is used. A mixture of these may be used. A preferred extraction solvent is water or a mixed solvent of water and alcohol.
Extraction is performed by room temperature extraction, heating extraction, or pressure extraction (autoclave treatment) or the like. Generally, it is performed at room temperature to 60 ° C. After the extraction treatment, the solid content and the liquid are separated by centrifugation or the like, and further subjected to a treatment such as filtration as necessary, followed by concentration by vacuum concentration or the like. Furthermore, it can also be pulverized by vacuum drying, freeze drying or the like. Appropriate excipients may be added during powdering.
[0012]
The active fraction can be obtained by membrane treatment (ultrafiltration, reverse osmosis, etc.) and fractionation by various chromatographies. Furthermore, the polyphenol fraction is purified by treating the extract with a membrane (ultrafiltration, reverse osmosis, etc.) and an adsorbent. As the adsorbent, styrene-divinylbenzene adsorbent, methacrylic acid adsorbent, hydrophilic vinyl polymer, modified dextran gel, polyacrylamide gel, reverse phase silica gel, ion exchange resin and the like are used. When these adsorbents are used, polyphenol is contained in a fraction adsorbed thereto (hereinafter referred to as an adsorbed fraction). By eluting the adsorbed fraction with hydrous alcohol, alcohol, acetone or the like, a purified polyphenol fraction can be obtained.
[0013]
In general, since low molecular weight components are more easily absorbed in the living body, fractionation was performed by ultrafiltration or the like, and high activity was observed in fractions having a molecular weight of 10,000 or less. Furthermore, an active extract can be obtained by performing an enzyme treatment or the like during extraction.
[0014]
The above-described processed products and fractions of buckwheat husks produced according to the present invention are administered to mice to cause in vivo substances caused by reactive oxygen species in the brain, heart, blood and other organs. Diseases resulting from oxidative degeneration are suppressed. Moreover, especially the effect | action which suppresses the production | generation of a lipid peroxide among these is shown. Moreover, it was confirmed that administration to mice reduces serum cholesterol and neutral fat, which were increased by ingestion of a high fat diet (a diet high in olive oil and cholesterol).
[0015]
Therefore, the buckwheat husk extract and fraction according to the present invention are useful as lipid peroxide inhibitors, cholesterol lowering agents, neutral fat lowering agents or hyperlipidemia improving agents.
Furthermore, peroxidic substances generated in vivo are one cause of degenerative changes that lead to aging, and suppressing the production of lipid peroxides, that is, exhibiting an antioxidant effect in vivo, It will prevent damage to furniture and aging. In particular, along with the aging society, cerebral dysfunctions such as dementia are increasing in recent years due to aging of the brain.
[0016]
In addition, these synergistic actions are expected to improve and prevent brain diseases such as stroke, cerebral dementia, Alzheimer's disease, cerebral infarction, cerebrovascular disorder, memory disorder and the like. In addition, in the blood system, improvement and prevention of adult diseases such as hypercholesterolemia, hyperlipidemia, arteriosclerosis, hypertension, ischemic diseases, and various organ disorders are expected. The
[0017]
In addition, the extract obtained from the buckwheat husk according to the present invention, the component having a molecular weight of 10,000 or less obtained by fractionating and purifying the same, and the polyphenol fraction obtained by further fractionating the extract are not toxic, As a result, it was confirmed that it can be used safely enough.
[0018]
In order to use an extract obtained from the buckwheat husk according to the present invention, a component having a molecular weight of 10,000 or less obtained by fractionating and purifying the same, and a polyphenol fraction obtained by further fractionating the same for the above purpose, Administration is systemic or local, in oral or parenteral form, with oral administration being preferred.
The dosage varies depending on age, body weight, symptoms, therapeutic effect, administration method, treatment time, etc., but is usually in the range of 100 mg to 600 mg per adult per dose.
When the compound of the present invention is administered, it is used as a solid composition for oral administration, a liquid composition and other compositions, an injection for parenteral administration, and the like.
Solid compositions for oral administration include tablets, pills, capsules, powders, granules, syrups and the like.
[0019]
【Example】
Next, although the manufacture example and test example of an extraction component are given and this invention is demonstrated further more concretely, this invention is not limited within these ranges.
Production example: Production method of buckwheat husk extract and production of fractions thereof After adding 4 liters of water and 4 liters of ethanol to 1 kg of buckwheat husk after obtaining buckwheat flour from Hokkaido soba seeds, extraction at 50 ° C. for 4 hours Was done. The obtained extract was filtered, concentrated under reduced pressure and dried to obtain 60.5 kg of buckwheat husk extract.
Among these, 10 kg of extract was dissolved in 500 ml of water, and fractionated by adding water appropriately with an ultrafiltration membrane (Advantech Ultra Filter Q100, molecular weight cut off 10,000), and after removing the fraction having a molecular weight of 10,000 or more, The obtained fraction was concentrated under reduced pressure to obtain 3.5 g of a fraction.
Further, 10 g of this extract was dissolved in 20 ml of water and fractionated with Sephadex LH-20 (manufactured by Formacia, modified dextran gel) to obtain 3.8 g of a polyphenol fraction adsorbed thereto. The obtained extract and fraction were used for the following tests.
[0020]
Test Example 1: Antioxidation of the above extract and fractions in the brain and other organs ddY male mice 8 weeks old, group of 5 mice, control, oat shell extract, fraction with a molecular weight of 10,000 or less And divided into 4 groups of polyphenol fractions. For the control, a normal diet was administered, and for the other diets, 1% each was mixed with the diet for 1 week. Mice were dissected 8 days after the start of administration, and blood, brain and heart were removed and used as samples. About each sample, the amount of lipid peroxide and the amount of oxidative modification substance were measured.
[0021]
The amount of lipid peroxide was measured by a method for determining the amount of lipid peroxide that causes each sample to react with 2-thiobarbituric acid (hereinafter abbreviated as TBA).
Specifically, 1 ml of physiological saline, 0.05 ml of blood or 100 μl of a homogenized sample was placed in a test tube and centrifuged at 3000 rpm for 10 minutes. To 0.5 ml of this supernatant, 4.0 ml of N / 12 sulfuric acid was added and stirred. To this was added 0.5 ml of 10% aqueous phosphotungstic acid solution, and the mixture was left standing for 5 minutes. This was centrifuged at 3000 rpm for 10 minutes, and 2.0 ml of N / 12 sulfuric acid was again added to the resulting precipitate and stirred. Then, 0.3 ml of 10% phosphotungstic acid aqueous solution was added and stirred, and then this was centrifuged at 3000 rpm for 10 minutes. A precipitate was obtained. After adding 4.0 ml of water to this precipitate and suspending it, 1.0 ml of TBA reagent (2-thiobarbituric acid 3.35 mg / ml, acetic acid 8.8 mol / l) was added and heated on a boiling water bath for 60 minutes. Cooled with. Add 5.0 ml of n-butanol to the obtained reaction product, mix well, extract the reaction product into a butanol layer, centrifuge at 3000 rpm for 10 minutes, and excite wavelength 515 nm, fluorescence wavelength for the upper butanol layer Fluorescence was measured at 553 nm. The measurement result about each sample of buckwheat husk extract, a fraction with a molecular weight cut-off of 10,000 or less, a polyphenol fraction, and a control is shown in FIGS. The vertical axis in the figure is the fluorescence intensity, which is shown on an arbitrary scale so that the relative intensity to the control can be understood. FIG. 1 shows the fluorescence intensity of the reaction product of lipid peroxide and TBA reagent in the blood, FIG. 2 shows the fluorescence intensity of the reaction product of lipid peroxide and TBA reagent in the brain, and FIG. 3 shows the lipid peroxide in the heart. The fluorescence intensity of the reaction product of the TBA reagent is shown.
[0022]
As apparent from FIGS. 1 to 3, compared with the control group, the oat husk extract administration group, the fractional molecular weight 10,000 or less administration group, and the polyphenol fraction administration group all have lipid peroxides in blood, brain, and heart. Controlled generation. In particular, it was remarkably observed in blood and brain.
The amount of the oxidatively modified substance was measured using the Bligh-Dyer method. The Bligh-Dyer method is a method in which a fat-soluble fluorescent substance produced by oxidative modification with active oxygen species in a living body is extracted and its fluorescence intensity is directly measured. That is, after homogenizing each sample with a phosphate buffer, extracting the fluorescent substance and removing the interfering substance, the obtained oxidatively modified substance is obtained using a fluorescence method.
[0023]
For brain and heart samples, 4 times the weight of phosphate buffer (pH 7.4) was added. Further, a blood sample was prepared by adding 2450 μl of physiological saline to 50 μl of blood, mixing and centrifuging at 3000 rpm for 10 minutes, and adding 1 ml of distilled water to the resulting precipitate. To 200 μl of the organ homogenization solution and the whole blood sample, 4 ml of a chloroform-methanol mixed solution (1: 2) was added and centrifuged at 3000 rpm for 10 minutes. Collect 3.5 ml of this supernatant, add 3 ml of chloroform and 6 ml of distilled water, mix and centrifuge at 3000 rpm for 10 minutes, collect the lower layer, add 3 ml of distilled water, mix and centrifuge at 3000 rpm for 10 minutes, After irradiating with ultraviolet rays for 1 minute, the lower layer was measured for fluorescence at an excitation wavelength of 357 nm and a fluorescence wavelength of 428 nm. The results are shown in FIGS. The vertical axis | shaft in a figure is the fluorescence intensity shown by the arbitrary scale so that the relative intensity | strength with control could be understood similarly to FIGS. 4 shows the fluorescence intensity of the oxidatively denatured substance in blood, FIG. 5 shows the fluorescence intensity of the oxidatively denatured substance in the brain, and FIG. 6 shows the fluorescence intensity of the oxidatively denatured substance in the heart.
[0024]
As apparent from FIGS. 4 to 6, compared to the control group, the oat husk extract administration group, the fractional molecular weight 10,000 or less administration group, and the polyphenol fraction administration group are blood, brain and heart in any fraction. The production of oxidatively denatured substances was suppressed. In particular, it was remarkably observed in blood and brain.
[0025]
Test Example 2: Serum cholesterol lowering action and triglyceride reducing action ddY male mice 8 lap ages of 5 mice per group, control, buckwheat shell extract, fraction with a molecular weight of 10,000 or less, and polyphenol fraction Divided into 4 groups. First, in all groups, olive oil and cholesterol mixed in a diet (high fat diet) were administered for 2 weeks, then the control diet was fed with a normal diet, and the others were 1% each. What was mixed with the normal diet at a ratio was administered for 1 week. On day 8 from the start of sample administration, mice were dissected, blood was collected, and serum cholesterol levels (total cholesterol) and triglycerides (triglycerides) were measured. The measurement results (concentration: mg / ml) for each sample of buckwheat husk extract, fraction with a molecular weight of 10,000 or less, polyphenol fraction, and control are shown in FIG. 7 and FIG. FIG. 7 shows the amount of cholesterol in the blood, and FIG. 8 shows the amount of triglyceride in the blood.
[0026]
As apparent from FIG. 7 and FIG. 8, the effect of lowering the amount of increased cholesterol is observed in the oat shell extract extract administration group, the fraction molecular weight fraction of 10,000 or less and the polyphenol fraction administration group.
[0027]
【The invention's effect】
The present invention is a lipid peroxide inhibitor, a cholesterol-lowering agent, comprising as an active ingredient an extract obtained by subjecting buckwheat husks that have been conventionally discarded and not used as a food or food additive, and an appropriately fractionated fraction, The present invention provides a neutral fat lowering agent and a hyperlipidemia improving agent.
The raw material of buckwheat husk can be supplied stably, and together with the development of a new application of utilizing unused resources, the present invention contributes significantly to the industry.
According to the present invention, it is possible to produce a large amount of an extract exhibiting effects such as excellent oxidative denaturation inhibition and lipid peroxidation inhibition from buckwheat husk. The extract according to the present invention can be safely used as a food, health food or food additive.
[Brief description of the drawings]
FIG. 1 is a graph comparing the fluorescence intensities of reaction products of lipid peroxide and TBA reagent in blood for mice administered with buckwheat husk extract, a fraction with a molecular weight of 10,000 or less, and a polyphenol fraction. is there.
FIG. 2 is a graph comparing the fluorescence intensities of reaction products of lipid peroxide and TBA reagent in the brain for mice administered with buckwheat husk extract, a fraction with a molecular weight of 10,000 or less, and a polyphenol fraction. is there.
FIG. 3 is a graph comparing the fluorescence intensities of reaction products of lipid peroxide and TBA reagent in the heart for mice administered with buckwheat husk extract, fractions with a molecular weight of 10,000 or less, and polyphenol fractions. is there.
FIG. 4 is a graph comparing the fluorescence intensity of oxidatively modified substances in blood for mice administered with a buckwheat husk extract, a fraction with a molecular weight cut-off of 10,000 or less, and a polyphenol fraction.
FIG. 5 is a graph comparing the fluorescence intensity of oxidatively modified substances in the brain of mice administered with a buckwheat husk extract, a fraction with a fractional molecular weight of 10,000 or less, and a polyphenol fraction.
FIG. 6 is a graph comparing the fluorescence intensity of oxidatively modified substances in the heart of mice administered with a buckwheat husk extract, a fraction with a fractional molecular weight of 10,000 or less, and a polyphenol fraction.
FIG. 7 is a graph comparing blood cholesterol levels (mg / ml) for mice administered with buckwheat husk extract, a fraction with a molecular weight cut-off of 10,000 or less, and a polyphenol fraction.
FIG. 8 is a graph comparing the amount of triglyceride in blood (mg / ml) for mice administered with a buckwheat husk extract, a fraction with a fractional molecular weight of 10,000 or less, and a polyphenol fraction.

Claims (6)

  A cholesterol-lowering agent, neutral fat-lowering agent, or hyperlipidemia-improving agent comprising, as an active ingredient, an extract obtained by extracting oat shells with water or an organic solvent alone or a mixture thereof.   The cholesterol-lowering agent, neutral fat-lowering agent, or hyperlipidemia-improving agent according to claim 1, comprising as an active ingredient a component having a molecular weight of 10,000 or less obtained by fractionating and purifying an extract obtained by extracting the buckwheat husk.   The cholesterol-lowering agent, neutral fat-lowering agent, or hyperlipidemia-improving agent according to claim 1, comprising a polyphenol fraction obtained by fractionating and purifying an extract obtained by extraction treatment of buckwheat husk as an active ingredient.   The cholesterol-lowering agent according to any one of claims 1 to 3.   The neutral fat lowering agent according to any one of claims 1 to 3.   The hyperlipidemia improving agent according to any one of claims 1 to 3.

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