JP2000072687A - Alpha-glucosidase inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an inhibitor easy to be taken in and having strong α- glucosidase inhibitory activity. SOLUTION: This α-glucosidase inhibitor includes Tohchi as an active ingredient. The Tohchi is generally sold as a food material for Chinese dishes and is obtained by fermenting steam boiled soybeans with Aspergillus. Daitoku temple natto (fermented soybeans produced in Kyoto city and colored black), Hama natto, temple natto and Ikkyu temple natto also belong to the Tohchi.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an .alpha.-glucosidase inhibitor comprising as an active ingredient a torch which can be used for pharmaceuticals, foods, health foods, foods for specified health use and the like.

[0002]

BACKGROUND OF THE INVENTION α- glucosidase inhibitors, inhibit α- glucosidase localized in microvilli of the small intestine, to suppress spikes and elevated insulin levels subsequent postprandial blood glucose, Diabate Medicine, 10 , 688 (1993). In humans and non-human animals, it shows an inhibitory effect on carbohydrates (especially, starch-derived oligosaccharides, sucrose, etc.). And it is useful for improvement of various diseases such as obesity and diabetes caused by hyperglycemia. In addition, (healthy) foods produced by adding an α-glucosidase inhibitor are suitable for a diet for patients with metabolic disorders, and are also suitable for healthy persons as a diet for preventing metabolic disorders.

[0003] As an α-glucosidase inhibitor derived from food, for example, Japanese Patent Application Laid-Open No. 9-65836 discloses
An enzyme hydrolyzate of an animal protein or a vegetable protein is disclosed, and Japanese Patent Application Laid-Open No. 5-17364 discloses a tea polyphenol.

[0004]

However, the α-glucosidase inhibitor described in Japanese Patent Application Laid-Open No. 9-65836 must be ingested in large quantities as a food in order to exhibit its activity. According to the technology disclosed in Japanese Patent Application Laid-Open No. Hei 5-17364, purification of polyphenol is complicated, and it is necessary to ingest a large amount of tea which is usually taken. In the present invention, α-glucosidase inhibitory activity is strong,
An object of the present invention is to provide an excellent α-glucosidase inhibitor which is easy to ingest.

[0005]

Means for Solving the Problems As a result of intensive studies, the present inventor has found that touchi (bean drum), which is also used as an ingredient in Chinese cuisine, has a strong α-glucosidase inhibitory activity. The present invention has been completed.

The torch of the present invention is generally commercially available as an ingredient for Chinese cuisine, and is usually made by fermenting steamed soybeans with koji, and is used to make natto from Daitokuji natto, Hama natto, Tera natto, and salt. Also, natto such as Ikkyuji natto is included in the present invention.

[0007] The torch is used in a usual form, and it is further dried to be powdered or pulverized in water to form a slurry, but shows glucosidase inhibitory activity.
Alternatively, an extract with an alcohol is preferable because a stronger α-glucosidase inhibitory activity is obtained, and an extract with a mixture of water and an alcohol is more preferable. Examples of the alcohol include methanol, ethanol, propanol, and butanol, but ethanol is preferred. As the extraction method, an extraction method such as heat extraction, continuous extraction, immersion, and supercritical extraction may be used. The mixing ratio (weight ratio) of water / alcohol is preferably from 1/100 to 100/1, and more preferably from 1/50 to 50/1.

More specifically, as an extraction method, a torch is crushed, and this powder is immersed in an equal weight mixture of water and alcohol (weight ratio of water / alcohol = 1/1) and heated.
By boiling for 0 to 30 minutes, the active ingredient can be extracted. In addition, the active ingredient can be extracted by immersing the torch in an equal weight mixture of water and alcohol and leaving it at room temperature for 5 hours to 7 days, or heating it at 40 to 60 ° C. for about 12 to 18 hours. .

The above-mentioned water and / or alcohol extract is
If water and / or alcohol is distilled off, a powder is obtained, which is preferably used as an α-glucosidase inhibitor. Further, in order to improve the effect of small ingestion, the extract is methanol, ethanol, propanol, butanol, chloroform, ethyl acetate, toluene, hexane,
The active ingredient is concentrated by a solvent fractionation operation using a polar organic solvent such as benzene, and then further subjected to alumina column chromatography, silica gel column chromatography, gel filtration chromatography, ion exchange chromatography, hydrophobic chromatography, and high performance liquid chromatography. Purification can also be carried out by appropriate separation and purification such as described above, or by combining one or several kinds.

[0010] The α-glucosidase inhibitor of the present invention has a blood glucose elevation inhibitory action, and is therefore suitable for liquid carriers such as water, ethanol, ethylene glycol and polyethylene glycol, and solid carriers such as starch, cellulose and polyamide powder. Diluted with a non-toxic carrier, and used as pharmaceuticals such as ampoules, granules, tablets, pills, capsules and syrups, as health foods, as a diet or prophylactic for patients with metabolic disorders, as a preventive for diabetes and obesity Can be. Furthermore, by taking the above-mentioned preparation containing the α-glucosidase inhibitor of the present invention before, during, after or between meals, an increase in blood glucose concentration due to eating can be suppressed.

The amount of human consumption at this time is preferably 0.1 to 50 g / day, particularly preferably 0.5 to 10 g / day, as a dry powder of torch. When using water and / or alcohol extracts or the above-mentioned isolated compounds, the intake may be 1/1000 to 1/10 of the dry powder.

The α-glucosidase inhibitor of the present invention can be added to foods, for example, coffee, fruit juice,
Soft drink, beer, milk, miso soup, soup, tea,
Liquid (fluid) foods such as tea, nutrients, syrup, margarine, jam, rice, bread, potato products, rice cake,
It can also be added to staple foods, side dishes, confectionery and seasonings such as candy, chocolate, sprinkle, ham, sausage, candy and other solid foods. The addition amount at this time is preferably 0.001 to 85% by weight, particularly preferably 0.01 to 60% by weight, as a dry powder of torch, based on the food. When using water and / or alcohol extract, 1/1000 of the dry powder
The addition amount may be 1/10 (weight ratio). In addition, in the case of foods, the intake amount is preferably the same as the above-mentioned pharmaceuticals and health foods, and furthermore, a sweetener, a preservative, a dispersant, a coloring agent, an antioxidant and the like within a range that does not inhibit the effects of the present invention. Can also be used in combination. Further, it may be used in combination with other known α-glucosidase inhibitors such as varienamine and aminocyclitol which are α-glucosidase inhibitors.

[0013]

The present invention will be specifically described below. still,
In the following description, "%" means% by weight. Example 1 1 kg of dried touch was immersed in 10 l of a 50% aqueous methanol solution for 1 day, and the obtained immersion liquid was concentrated by a rotary evaporator, dissolved in water, and filtered.
The obtained filtrate was concentrated under reduced pressure to obtain an extract (E-1).
The extract was evaporated to dryness on a rotary evaporator and 193
g of solid was obtained. The solid was put into a separatory funnel, and a mixed solution of ethyl acetate / water (volume ratio = 1/2) was mixed with one part of the solid.
The aqueous layer (E-2) and the ethyl acetate layer (E-3) were fractionated by adding 10 to 10 times the weight, and the α-glucosidase inhibitory activity was measured. Each fraction (E-1, E-
2. The α-glucosidase inhibitory activity of E-3) was measured as follows.

(1) Preparation of di (tri) sugar hydrolase (α-glucosidase) from rat small intestine The frozen small rat intestine (jejunum) was thawed, and the mucous membrane was extruded using forceps. . A 0.1 M potassium phosphate buffer (pH 7.0) containing 5 times the weight of 5 mM ethylenediaminetetraacetic acid was added to the mucous membrane, and homogenized while cooling. Thereafter, centrifugation (4 ° C., 21000
× g, 60 minutes), and a 0.1 M potassium phosphate buffer (pH 7.0) containing 1% Triton X-100 was added to the obtained precipitate so as to have a 5-fold weight, and solubilization treatment (4 ℃,
60 minutes). This is ultracentrifuged (4 ° C, 1100
00 × g, 90 minutes), and the supernatant is dialyzed against 0.01 M potassium phosphate buffer (pH 7.0) (4 ° C., 24 hours)
And used as an enzyme solution.

(2) Enzyme (α-glucosidase) activity
Measurement Enzyme activity was measured using a commercially available kit.
Source was used. Standard reaction solution composition is 60 mM substrate solution
(Sucrose in 0.1 M potassium phosphate buffer pH
0.7 ml, dissolved in 6.3), test substance solution
(The water and organic solvent of each fraction were completely removed.
And then dissolved in a 50% aqueous solution of dimethyl sulfoxide) 0.2
ml, and the above enzyme solution was 0.1 ml (1.0 ml in total).
This was reacted at 37 ° C. for 15 minutes, and then buffered with 2 M Tris-HCl buffer.
Stop the reaction using 1.5 ml of the solution (pH 7.0)
The test solution was used. The concentration of the test substance in the reaction solution was 20 mg.
/ Ml. Next, one hole was placed on a 96-well microplate.
Recoloring reagent [Glucose B Test Wako (Wako Pure Chemical Industries, Ltd.)
50 μl of test solution in 200 μl (ethyl acetate, etc.
Removed) and incubate at 37 ° C for 30 minutes
And then a microplate reader (BIO RAD)
Absorbance at 490 nm with Model 550)
Was. 0.1 M potassium phosphate buffer instead of substrate solution
The absorbance when (pH 6.3) was added was taken as a blank value,
The value obtained by subtracting this value is A_490sAnd Instead of test liquid
Absorption when adding 50% by weight aqueous solution of dimethyl sulfoxide
Control the brightness (A _490c), And α-
Glucosidase inhibitory activity was determined. The measurement is performed twice,
The average was taken as the measured value.

[0016]

Α-glucosidase inhibitory activity (%) = {(A _490c
−A _490s ) / A _490c } × 100 The α-glucosidase inhibitory activity of each fraction (E-1, E-2, E-3) is shown in Table 1.

[0017]

[Table 1] *: Each fraction was concentrated to dryness to determine the weight,
The value was based on the weight of the dried raw material.

[0018]

Industrial Applicability The glucosidase inhibitor of the present invention contains torch as an active ingredient and is easy to ingest and exhibits a strong inhibitory activity.

Claims (2)

[Claims] 1. An α-glucosidase inhibitor comprising touchi as an active ingredient. 2. The α-glucosidase inhibitor according to claim 1, wherein a water and / or alcohol extract of the torch is used as the torch.