JP2000229873A - Alpha-glucosidase inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject inhibitor having strong α-glucosidase inhibitory activity, being readily taken by making the inhibitor include a plant belonging to the family Rosaceae as an active ingredient. SOLUTION: This inhibitor contains a plant belonging to the family Rosaceae as an active ingredient. For example, wild strawberry, rose, Chrysosplenium grayanum, Hydrangea paniculata, Rubus phoenicolasius, etc., may be cited as the plant belonging to the family Rosaceae. Though the plant belonging to the family Rosaceae is used in an ordinary state, further dried and powdered or ground in water into a slurried state and the slurried plant may show glucosidase inhibitory activity, an extract obtained by using water and/or an alcohol preferably provides stronger α-glucosidase inhibitory activity. An extract obtained by using a mixed solution of water and an alcohol is especially preferable. Ethanol is preferable as the alcohol. The weight ratio of water/the alcohol is preferably 1/100 to 100/1, more preferably 1/50 to 50/1.

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 containing as an active ingredient a plant belonging to the family Rosaceae, which can be used in medicines, foods, health foods, foods for specified health use, and the like. is there.

[0002]

2. Description of the Related Art An α-glucosidase inhibitor inhibits α-glucosidase localized in microvilli of the small intestine, and suppresses a rapid increase in blood glucose level after meal and a subsequent increase in insulin level. , 688 (1993). In humans and non-human animals, it inhibits the metabolism of carbohydrates (especially starch-derived oligosaccharides and sucrose). 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. An object of the present invention is to provide an excellent α-glucosidase inhibitor which has a strong α-glucosidase inhibitory activity and is easy to ingest.

[0005]

As a result of intensive studies, the present inventors have found that a plant belonging to the family Rosaceae has a strong α-glucosidase inhibitory activity, and have completed the present invention.

[0006]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described below in detail.

[0007] The plants belonging to the Rosaceae of the present invention include wild strawberry, rose, cat flea, squirrel,
Ebigarai strawberry, Ezoyamazakura, Chishimanekome, Hirohanokawarasaiko, Watageka matsuka, Triassium terrestris, and the like.

[0008] The plants belonging to the family Rosaceae can be used in a usual form. Further, they can be dried and pulverized, or crushed in water to be slurried to show glucosidase inhibitory activity. Is preferable since a stronger α-glucosidase inhibitory activity is obtained, and an extract in a mixed solution of water and alcohol is particularly 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 extraction, or supercritical extraction may be used. When water and alcohol are used in combination, 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.

Specifically, as an extraction method, a dried plant belonging to the family Rosaceae is pulverized (dried), and this powder is mixed with an equal weight of a mixture of water and alcohol (weight ratio of water / alcohol = 1/1). ), Heated and boiled for 10 to 30 minutes to extract the active ingredient. Also, the plant is mixed with an equal weight of a mixture of water and alcohol (water / water).
(Weight ratio of alcohol = 1/1), at room temperature for 5 hours
The active ingredient can be extracted by leaving it to stand for 7 days or heating it at 40 to 60 ° C. for about 12 to 18 hours.

The above 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 an organic solvent such as benzene, and further subjected to alumina column chromatography, silica gel column chromatography, gel filtration chromatography, ion exchange chromatography,
Purification can also be carried out by appropriate separation and purification such as hydrophobic chromatography and high performance liquid chromatography by combining one or more kinds.

Since the α-glucosidase inhibitor of the present invention has a blood glucose elevation inhibitory effect, it can be used 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.

At this time, the amount of human intake is 0.1 to 50 g / (dry plant) as the dry powder.
Days are preferred, especially 0.5 to 10 g / day. When a water and / or alcohol extract or the extract described above is used, 1/1000 to 1/1 of the dry powder
It may be 0 intake.

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 solid foods such as candy, chocolate, sprinkle, ham, sausage, and candy. At this time, the added amount is 0.001 to 85% by weight based on the above-mentioned food as dry powder of the plant (belonging to Rosaceae)
Is preferred, and particularly preferably 0.01 to 60% by weight. When water and alcohol extracts are used, 1 /
The addition amount may be 1000 to 1/10. In addition, in the case of food, the intake amount is preferably the same as that of the above-mentioned medicines 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.

[0014]

The present invention will be specifically described below. still,
In the following description, "%" means% by weight. Example 1 1 kg of wild strawberry was immersed in 1 liter 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. The extract was dried using a rotary evaporator to obtain 380 g of a solid. The solid was placed in a separatory funnel, and hexane was added at 30 times the weight of the solid to fractionate into a hexane layer (E-1) and an aqueous layer. The layer (E-2) was extracted. Each fraction (E
-1, E-2) α-glucosidase inhibitory activity was measured as follows.

(1) Preparation of Enzyme Solution The small intestine (jejunum) of the frozen rat was thawed, and the mucous membrane was collected by extruding it with 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)
Then, an enzyme (α-glucosidase) solution was obtained.

(2) Measurement of α-glucosidase activity A commercially available kit was used for α-glucosidase activity, and sucrose was used as a substrate. The standard reaction solution composition is 60
0.7 ml of a mM substrate solution (sucrose dissolved in 0.1 M potassium phosphate buffer, pH 6.3) and a test substance solution (E obtained from 0.1 g of wild strawberry)
-1 and E-2 fractions were concentrated to dryness and then dissolved in a 50% aqueous dimethyl sulfoxide solution so as to have a volume of 1 ml each) 0.2 ml, and the above enzyme solution 0.1 ml (1.0 m in total)
l). This was reacted at 37 ° C. for 15 minutes, and the reaction was stopped with 1.5 ml of 2M Tris-HCl buffer (pH 7.0) to obtain a test solution. Next, 50 μl of a test solution was added to 200 μl of a coloring reagent [Glucose B Test Wako (manufactured by Wako Pure Chemical Industries)] per well in a 96-well microplate.
After incubation at 30 ° C. for 30 minutes, the plate was incubated for 4 minutes with a microplate reader (BIO RAD, MODEL550).
The absorbance at 90 nm was measured. 0.1 instead of substrate solution
The absorbance when the M potassium phosphate buffer (pH 6.3) was added was used as a blank value, and the value obtained by subtracting this value was used as A _{490 s}
And The absorbance when a 50% by weight aqueous solution of dimethyl sulfoxide was added instead of the test solution was taken as a control value ( _A490c ), and the α-glucosidase inhibitory activity was determined by the following formula. The measurement was performed twice, and the average value was used as the measured value.

[0017]

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

[0018]

[Table 1]

[0019]

Industrial Applicability The glucosidase inhibitor of the present invention contains a plant belonging to the family Rosaceae as an active ingredient, so that it is easy to ingest and exhibits a strong inhibitory activity.

Claims (3)

[Claims] An α-glucosidase inhibitor comprising a plant belonging to the family Rosaceae as an active ingredient. 2. The α-glucosidase inhibitor according to claim 1, wherein an extract obtained by extracting a plant belonging to the family Rosaceae with water and / or alcohol is used as an active ingredient. 3. The α-glucosidase inhibitor according to claim 1, wherein the plant belonging to the family Rosaceae is wild strawberry.