KR20100078830A - THE EXTRACTS FROM VIGNA SP. COMPRISING α-GLUCOSIDASE INHIBITORY ACTIVITY - Google Patents

Abstract

PURPOSE: A Vigna sp. extract having activity of suppressing alpha-glucosidase is provided to ensure blood glucose reductin, anti-obesity, and anti-oxidation. CONSTITUTION: A method for manufacturing Vigna sp. extract having an effect of suppressing alpha-glucosidase comprises: a step of pulverizing seed of Vigna; a step of adding 1-6 low alcohol or solution thereof and stirring at 20-30 °C to extract; a step of centrifuging extraction liquid to collect supernatant; and a step of performing decompression or freeze drying to obtain light brown and dark brown.

Description

Extract of red bean cultivars with alpha-glucosidase inhibitory activity {The extracts from Vigna sp. comprising α-glucosidase inhibitory activity

The present invention relates to the use of the red bean myrtle seed extract having alpha-glucosidase inhibitory activity.

Vigna sp. Crops are wild or cultivated in the mountains and mountains of Korea. Red beans related crops include red beans ( Vigna angularis ), green beans ( Vigna radiata ), red beans ( Vigna umbellata ), red beans ( Vigna nakashimae) and Vigna angularis var.nipponensis. In particular, red beans ( Vigna angularis ) is known to be effective in kidney disease, stool, tumor, maternity disease since ancient times, it is also used as a folk remedy. Red bean saponins (DDMP-saponins) have antioxidant and free radical scavenging activity and hydrothermal extracts are known to stimulate tyrosinase activity and inhibit hair whitening. In addition, Vigna radiata has been used for antipyretic, detoxification, boils, swelling, and rheumatism, and related products for skin beauty using vitexin and isobitexin in mung beans have been developed and sold. It is becoming. Like this, it is a crop that genetically contains various bioactive substances of R. aeruginosa crops and has a great agricultural and industrial value by scientifically proving various biological activities.

However, red bean red bean , new red bean And Red bean It is not disclosed that the extract has a hypoglycemic effect by inhibiting the action of alpha-glucosidase. In addition, there has not been disclosed a method for producing an extract containing a substance that inhibits alpha-glucosidase activity from these plants or seeds.

The average life expectancy has been prolonged due to the changes in diet and lifestyle caused by the rapid economic development of modern society, while chronic degenerative diseases such as diabetes are increasing. In Korea, 4 million people, 8% of the population in 2004, were reported to have diabetes. In particular, dietary care is very important in the management of type 2 diabetic patients, and fundamentally to control hyperglycemia. If postprandial hyperglycemia is not controlled, it can promote vision disorders, neurological disorders, renal and cardiac dysfunction due to chronic diabetic complications of retinopathy. Therefore, people with diabetic disease should be absorbed slowly carbohydrates of the ingested food to prevent the rapid rise in blood sugar. Alpha-glucosidase inhibitors are drugs that have been developed and used to address this problem. Alpha-glucosidase is an enzyme that hydrolyzes digested oligosaccharides into monosaccharides in the small intestine. Therefore, the development of this enzyme inhibitor will relieve carbohydrate uptake by delaying carbohydrate absorption, which will be helpful in treating diabetes. Thus, many studies have been made on the inhibition of alpha-glucosidase, which is essential for carbohydrate metabolism, for use in regulating blood sugar drop. In particular, in Bayer, Germany, in 1977 acabos [acaebos, Schmidt, D.D. Et al., Naturwis senschaften, 1977, 64, 535] and the deoxynojirimycin derivative, miglitol, has been developed and marketed for the treatment of diabetes and obesity.

To date, many types of anti-diabetics have been developed and are under development, but no satisfactory drugs have been developed. Currently, oral hypoglycemic drugs such as sulfonyl urea and meglitinide may cause hypoglycemia, colds, and headaches when taken with other drugs, and biguanide is a gastrointestinal disorder. And renal toxicity. Therefore, efforts are underway to develop alpha-glucosidase inhibitors for the treatment of diabetes that have relatively few side effects and can control blood glucose regulation.

Accordingly, the present inventors, while searching for an alpha-glucosidase inhibitor in the main grains of red bean seedlings in order to minimize the side effects of existing therapeutic agents, the extract of red bean seedlings inhibits alpha-glucosidase enzyme activity. The invention was first discovered and the present invention was completed by developing these extraction methods in a simple process.

Accordingly, it is an object of the present invention to provide an extract of rhododendron myomas having alpha-glucosidase inhibitory activity.

In addition, another object of the present invention is to provide a method for centrifugation and vacuum concentration after grinding with a lower alcohol by grinding adzuki bean seedling seed.

In addition, another object of the present invention is to provide a composition for preventing and treating diabetes mellitus, diabetic disease or obesity as the active ingredient of the rhododendron myoma.

In addition, another object of the present invention is to provide a health food for preventing and improving diabetes, diabetic disease or obesity using the adzuki bean jongjong extract as an active ingredient.

The present invention

1) pulverizing seeds of adzuki bean crops, extracting by stirring at 20 to 30 ℃ with a lower alcohol having 1 to 6 carbon atoms or an aqueous solution thereof having 10 to 20 times the weight of the seeds;

2) recovering only the supernatant by centrifuging the extract; And

3) concentrating the liquid under reduced pressure and lyophilization to prepare a light brown or brown powder

It characterized in that the preparation method and the adzuki bean jongjong extract prepared from the adzuki bean jongjong extract having an alpha-glucosidase inhibitory effect consisting of.

In addition, the pharmaceutical composition and health food containing red bean seedling extract as an active ingredient is another feature.

Hereinafter, the present invention will be described in detail.

In the present invention, Vigna sp. Extract exhibits a strong inhibitory effect of alpha-glucosidase, which has a hypoglycemic and obesity inhibitory effect, and has an antioxidant effect, thus preventing and treating diabetes mellitus, diabetic disease or obesity. The composition relates to medicines and health functional food ingredients.

Method for preparing the red bean root seedling extract according to the present invention is as follows.

First, the seeds of adzuki bean crops are crushed and extracted by stirring at 20 to 30 ° C. with a lower alcohol having 1 to 6 carbon atoms or an aqueous solution thereof having 10 to 20 times the weight of the seeds. The red bean seed crops include red beans ( Vigna angularis ), green beans ( Vigna radiata ), red beans ( Vigna umbellata ), red beans ( Vigna nakashimae) and Red bean ( Vigna angularis var.nipponensis ) is any one selected from the group consisting of.

Seeds of 100 adzuki bean seed crops were finely ground and used as analytical materials, and the yield was changed according to the extraction solvent conditions.

The extract is centrifuged to recover only the supernatant, concentrated under reduced pressure and lyophilized to give a light brown or brown powder.

The present invention is economical and convenient to provide an extract having excellent alpha-glucosidase inhibitory activity by only a simple process of pulverizing and extracting the adzuki bean cultivars seeds by shaking and extracting them with an alcohol or an aqueous solution thereof and depressurizing or lyophilizing the supernatant centrifuged. It is a way.

The rapid alpha-glucosidase inhibitory activity assay using adzuki bean cultivars extract was performed by ELISA method, and the concentration of alpha-glucosidase by concentration after stopping centrifugation and concentration of the sample for accurate activity assessment Inhibitory activity was evaluated. The substrate used for the evaluation of alpha-glucosidase inhibitory activity was para-nitrophenyl a-D-glucopyranoside, which was spectroscopy after reaction in 0.1 M phosphate buffer (pH 7.0). Analyzed by photometer (400 nm). Enzyme inhibitory activity was first compared to background experiments by enzymatic activity of 100% (based experiment) in the experiment with the same volume of solvent instead of the extract.

As described above, it was identified for the first time that the obtained adzuki bean ethanol extract has a strong inhibitory effect on alpha-glucosidase which is an essential enzyme in carbohydrate digestion and absorption in the human body.

Therefore, the present invention includes a diabetes mellitus, diabetic disease or obesity prevention and treatment composition comprising the extract of the rhododendron myoma.

The diabetic disease is a degenerative complication such as diabetic retinopathy, diabetic neuropathy, kidney disease, atherosclerosis, myocardial disorders, skin disorders, diabetic foot syndrome, and horse blood vessel disease as diabetes progresses chronically. Included.

In addition, the extract of the rhododendron rhododendron is an active ingredient formulation, for example, tablets, troches, lozenges, water-soluble or oily suspensions, powders, powders or granules, emulsions, hard or soft. It is formulated as a capsule, suspension, emulsion, syrup or elixirs. Binders such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose or gelatin for formulation into formulations such as tablets and capsules; Excipients such as dicalcium phosphate; Disintegrants such as corn starch or sweet potato starch; Lubricants such as magnesium stearate, calcium stearate, sodium stearyl fumarate or polyethylene glycol wax. In the case of the capsule formulation, in addition to the substances mentioned above, it contains a liquid carrier such as fatty oil. In addition, the active ingredient can be administered parenterally, parenteral administration is by subcutaneous injection, intravenous injection, intramuscular injection or intrathoracic injection injection method. To formulate into a parenteral formulation, the composition is prepared in solution by mixing in water with stabilizers or buffers and formulated into unit dosage forms of ampoules or vials.

The effective amount of the active ingredient may be varied depending on the age, physical condition, weight, etc. of the patient, but in general, it is preferable to administer within the range of 10 to 100 mg / kg (weight) / day. In addition, within a daily effective dosage range, it can be injected once or divided into several times a day.

On the other hand, red bean seedling extract of the present invention can be applied to health food.

In addition, the health food is a food prepared by adding the active ingredient to food materials such as beverages, teas, spices, gums, confectionery, capsules, powdered, suspensions, etc. Unlike the general medicine, it means that it is brought, but it is possible to take it as an anti-obesity food or an obesity treatment supplement because it has no merit such as side effects that may occur when taking the medicine for a long time.

The food composition of the present invention includes the red bean seedling extract as an active ingredient, but may be used in addition to other foods or food additives. The blending amount of the active ingredient with other additives may be appropriately determined within a conventional range depending on the purpose of use (prevention or treatment).

Red bean root jongjong extract may be included 0.01 to 100% by weight of the total food composition. However, the content in the case of long-term intake for the purpose of weight control and prevention of obesity may be below the above range, and since there is no problem in terms of safety and side effects, the active ingredient may be used in an amount above the above range.

There is no particular limitation on the kind of food to which the composition of the present invention is applied. Examples of the food to which the substance can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, drinks, Alcoholic beverages and vitamin complexes, and the like and include all of the health foods in the conventional sense.

The health beverage of the present invention may contain various flavors or natural carbohydrates and the like as additional ingredients, as in general beverages. The natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin and sugar alcohols such as xylitol, sorbitol and erythritol. As a sweetener, natural sweeteners, such as tautin and a stevia extract, or synthetic sweeteners, such as saccharin and aspartame, can be used. The proportion of the natural carbohydrate is generally from 0.01 to 0.04 g, preferably from 0.02 to 0.03 g per 100 ml of the composition of the present invention.

In addition to the above ingredients, the health food of the present invention may contain various nutrients, vitamins, electrolytes, flavors, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin, Alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the health food of the present invention may contain fruit flesh for the production of natural fruit juice, fruit juice beverage and vegetable beverage. These components can be used independently or in combination. The proportion of such additives is not critical but is generally selected in the range of 0.01 to 0.1 parts by weight with respect to 100 parts by weight of the composition of the present invention.

The preparation method of the red bean root seedling extract according to the present invention is an economical extraction method because the amount of the organic solvent is reduced and no separate 2-3-step purification process is required.

In addition, the red bean seedling extract according to the present invention inhibits the absorption of glucose in the blood by inhibiting alpha-glucosidase, inhibits the rise of blood sugar after eating, and inhibits less than that is a very useful invention in the biological and health food industry.

Therefore, the present invention has a high value for use as a medicine and functional food material as a composition for the prevention and treatment of diabetes mellitus, diabetic disease or obesity through a simple method of preparing zucchini extract having excellent alpha-glucosidase inhibitory activity, Is expected to increase value added in the future.

Hereinafter, the present invention will be described in detail by way of examples, but the scope of the present invention is not limited thereto.

Example 1 Preparation of Red Bean Seedling Extract

Seeds of red beans, mung beans, red beans, fresh red beans, and red bean red beans of the red bean seedling crops were ground to a fine uniform particle size with a high-speed vibratory grinder (HEICO, LT1-100, Japan), and then weighed 15 times per sample (kg). The mixture was extracted with stirring at room temperature with a corresponding 80% ethanol solvent. The extracted material was centrifuged at 3000 rpm to take only the supernatant, and then concentrated under reduced pressure at 40 ° C. or lower, and freeze-dried once more to give a light brown to brown powder [red bean extract 120g (yield 1.2%), mung bean extract 60g (yield 0.6). %), Red bean extract 80g (yield 0.8%), wild red bean extract 50g (yield 0.5%) and zodiac red bean extract 80g (yield 0.8%)] were obtained [Fig. 2].

The extraction solvent used in the present invention includes all methods including ethanol, methanol and its aqueous solution, and when the ethanol and methanol concentration is 50% or less, the extraction efficiency is significantly reduced to 50 to 100%.

In the present invention has the advantage that can be saved economically and timely as there is no need for an additional process of the second to third purification process of the existing extract.

Figure 1 shows the shape and characteristics of the zucchini seeds.

The average weight of the red bean seeds is 2.1g, and they have various species such as gray, brown and black.

Example 2: Determination of alpha-glucosidase inhibitory activity

In order to measure the alpha-glucosidase inhibitory activity of the seed extracts of the red beans, green beans, red beans, wild red beans, and red bean red beans obtained in Example 1, the enzyme alpha-glucosidase and para-nitrophenyl α- _D − as substrates were measured. Glucopyranoside ( para -nitrophenyl α- _D -glucopyranoside) was purchased from Sigma, and 0.1M phosphate buffer (pH 7.0) was used directly. 20 µl of rhododendron extract was added to 20 µl of 0.07 units / mL alpha-glucosidase, followed by 2910 µl of 0.1M phosphate buffer, followed by reaction at 37 ° C. for 20 minutes, followed by para-nitrophenyl α- _D- . 50 μl of glucopyranoside was added and the absorbance was measured at 400 nm. At this time, the final concentration of the substrate used was 0.4mM. The inhibition rate was calculated from the difference in absorbance with the reaction solution into which ethanol was added instead of the extract, and the results are shown in FIGS. 3A and 3B. Curcumin (curcumin) and zodiac extracts of the control also measured the inhibition rate in the same manner as above and the results are shown in Figure 3a. In addition, Equation 1 applied to the activity evaluation of the enzyme is as follows.

[Equation 1]

Alpha-glucosidase activity (%) = 100 [1 / (1 + ([extract concentration] / IC ₅₀ ))]

The IC ₅₀ indicates an extract concentration that inhibits alpha-glucosidase activity by 50%. As shown in FIG. 3a, the extracts of Zoldol red beans showed very strong inhibitory activity.

A strong inhibitory effect of inhibiting 50% of alpha-glucosidase activity at a concentration of 0.007 mg / mL of zodiac red beans extract was confirmed [FIG. 4]. The activity level of zucchini red bean extract was confirmed to be 2 times stronger than the inhibitory activity of curcumin (0.014 mg / mL) showing alpha-glucosidase activity among natural substances.

Example 3: Confirmation of Antioxidant Activity

Antioxidant activity was measured by DPPH radical scavenging method. That is, 1.0 g of red bean seedlings were extracted with 20 ml 80% ethanol, 50 μL of the extract was mixed with a final concentration of 100 μM DPPH and reacted for 30 minutes, and then measured by absorbance at 517 nm to evaluate its activity. As shown in Figure 5, Zamdol red bean extract of the red bean seed crops showed very strong antioxidant activity.

Example 4 Toxicity Test

Toxicity experiments were carried out on the extract of zodiac red beans, the active ingredient of the present invention as follows.

Zoldol red bean extract was dissolved in 1% sodium carboxymethly cellulose, diluted with water and orally administered to mice (10 per group) to 0, 4, 20, 100, 500 mg / kg, respectively, and then 7 Observed daily. No rats died due to oral administration of zodiac extract, and no rats died for 7 days.

Formulation Example 1 Preparation of Tablet

A tablet containing 15 mg of red bean extract as an active ingredient is prepared by the following method.

250 g of active ingredient was mixed with 175.9 g of lactose, 180 g of potato starch, and 32 g of colloidal silicic acid. 10% gelatin solution was added to the mixture, which was then ground and passed through a 14 mesh sieve. It was dried and the mixture obtained by adding 160 g of potato starch, 50 g of talc and 5 g of magnesium stearate was made into a tablet.

Formulation Example 2 Preparation of Capsule

500 mg of the active ingredient (mudstone red bean extract) was filled into hard gelatin capsules to prepare a capsule.

Formulation Example 3: Beverage Preparation

After dissolving 500 mg of the active ingredient (mudstone red bean extract) in an appropriate amount of water, vitamin C as an auxiliary component, citric acid, sodium citrate, oligosaccharides as an auxiliary component, and an appropriate amount of sodium benzoate as a preservative are added, followed by water. To 100 ml to prepare a beverage composition. At this time, taurine, myo-inositol, folic acid, pantothenic acid, etc. may be added alone or together.

Diabetes is a disease in which glucose in the blood is excreted in the urine. It is a chronic degenerative disease that cannot be cured fundamentally. 4 million people, 8% of Korea's population, have diabetes. These diabetics are afraid of rapid blood sugar rise after carbohydrate intake and take commercial oral hypoglycemic agents.

The present invention relates to the use of a method for preparing and extracting a zodiac red bean extract having a strong inhibitory effect on alpha-glucosidase. It has the advantage of reducing the economic and time effort consumed in the 2nd ~ 3rd purification process. It is possible to control the blood sugar immediately after meals by showing the strong inhibitory effect of alpha-glucosidase with only the extract of Zoldol adzuki bean, and the extract of the present invention has an anti-oxidant effect at the same time. And high functional value as a functional food material, zomdol red beans are expected to increase the added value in the future.

Figure 1 is a photograph showing the diversity of the shape of the seed and the seedlings of the rye crops.

2 is a graph illustrating a method of extracting the alpha-glucosidase inhibitory active ingredient from the seed of red bean red beans.

Figure 3a shows the degree of inhibition of alpha-glucosidase for adzuki bean crop crop extracts, Figure 3b is a graph showing the change in absorbance according to the concentration of red bean extract.

Figure 4 is a graph showing the alpha-glucosidase inhibitory effect of zodiac red bean extract.

Figure 5 is a graph showing the DPPH radical scavenging activity of the red bean seedling extract.

Claims (8)

1) pulverizing seeds of adzuki bean crops, extracting by stirring at 20 to 30 ℃ with a lower alcohol having 1 to 6 carbon atoms or an aqueous solution thereof having 10 to 20 times the weight of the seeds; 2) recovering only the supernatant by centrifuging the extract; And 3) concentrating the liquid under reduced pressure and lyophilization to prepare a light brown or brown powder Method of producing adzuki bean jongjong extract with an alpha-glucosidase inhibitory effect, characterized in that consisting of. According to claim 1, wherein the red bean seedling crops include Vigna angularis , Mung bean ( Vigna radiata ), Red bean ( Vigna umbellata ), Red beans ( Vigna nakashimae) and A method for producing red bean seedling extract, characterized in that any one selected from Vigna angularis var.nipponensis. According to claim 1, wherein the adzuki bean jongjong extract is an adzuki bean jongjong extract manufacturing method, characterized in that the extract. Red bean root seedling extract having an alpha-glucosidase inhibitory effect, which is prepared by the method of any one of claims 1 to 3. Diabetic, diabetic disease or obesity prevention and treatment composition, characterized in that the extract as an active ingredient. According to claim 5, wherein the red beans are related to red beans ( Vigna angularis ), green beans ( Vigna radiata ), red beans ( Vigna umbellata ), red beans ( Vigna nakashimae) and Composition comprising any one selected from Vigna angularis var.nipponensis. Health food for the prevention and improvement of diabetes mellitus, diabetic disease or obesity, characterized in that the extract of red bean root jongjong as an active ingredient. According to claim 7, wherein the red beans are related to red beans ( Vigna) angularis), mung bean (Vigna radiata), yepat (Vigna umbellata , Vigna nakashimae ) and Health food, characterized in that any one selected from Vigna angularis var.nipponensis.

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