JP2000229869A - α-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 Chinese herb as an active ingredient. SOLUTION: This inhibitor contains a Chinese herb as an active ingredient. For example, Magnolia bark, Artemisiae capillaris flos, Citrus Unshiu Peel, Evodia rutaecarpa, peppermint, Platycodon grandiflorum, Lycium chinense, Schizandra chinensis, etc., may be cited as the Chinese herb. Though the Chinese herb is used in an ordinary state, further dried and powdered or ground in water into a slurried state and the slurried Chinese herb 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. Methanol, ethanol, propanol, butanol, etc., may be cited as the alcohol and ethanol is preferable. The weight ratio of water/the alcohol is preferably 1/100 to 100/1.

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 a Chinese herb as an active ingredient, which can be used in medicines, foods, health foods, foods for specified health use and the like.

[0002]

2. Description of the Related Art An inhibitor of α-glucosidase inhibits α-glucosidase localized in microvilli of the small intestine and suppresses a rapid increase in blood glucose after meal and a subsequent increase in insulin. , 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. 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]

The present inventors have conducted intensive studies and as a result, Chinese herbs which are also used as pharmaceuticals, foods, health foods and foods for specified health use have a strong α-glucosidase inhibitory activity. Thus, the present invention has been completed.

[0006]

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

The Chinese herbs of the present invention include, for example, magnolia, inchenko, chenhi, goshuyu, sheik, kikyo, kyohime, omisho.

[0008] Chinese herbs can be used in the usual form, and further exhibit glucosidase inhibitory activity even when dried and pulverized or crushed in water to form a slurry, but extracted with water and / or alcohol. Things are stronger α-
Glucosidase inhibitory activity is preferred, and an extract in a mixture of water and alcohol is particularly preferred. Examples of the alcohol include methanol, ethanol, propanol, and butanol, but ethanol is preferred. Extraction methods include heating extraction, continuous extraction, immersion extraction,
An extraction method such as 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.

As an extraction method, specifically, Chinese herbs are crushed (dried), and this powder is immersed in an equal weight mixture of water and alcohol (weight ratio of water / alcohol = 1/1). By heating and boiling for 10-30 minutes,
The active ingredient can be extracted. Further, the food is immersed in an equal weight mixture of water and alcohol (weight ratio of water / alcohol = 1/1) and left at room temperature for 5 to 7 days, or heated at 40 to 60 ° C for about 12 to 18 hours. By doing so, the active ingredient can be extracted.

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 human intake is preferably 0.1 to 50 g / day as dry powder of Chinese herb.
Particularly, 0.5 to 10 g / day is preferable. When using a water and / or alcohol extract or the extract described above, 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 solid foods such as candy, chocolate, sprinkle, ham, sausage, and candy. The amount added at this time is preferably 0.001 to 85% by weight, and particularly preferably 0.01 to 60% by weight, as a dry powder of Chinese herbs, based on the food. When using water and / or alcohol extract, 1/1000 of the dry powder
The addition amount may be up 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, other known α-glucosidase inhibitors such as valienamin and aminocyclitol,
It may be used in combination with a glucosidase inhibitor.

[0014]

The present invention will be specifically described below. still,
In the following description, "%" means% by weight. Example 1 1 kg of magnolia 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 a solid. The solid was placed in a separatory funnel, and a mixed solution of ethyl acetate / water (volume ratio = 1/2) was added 1 to 10 times the weight of the solid, and the aqueous layer (E-
1) The ethyl acetate layer (E-2) was fractionated, and α
-The glucosidase inhibitory activity was measured. The α-glucosidase inhibitory activity of each fraction (E-1, E-2) 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 with tweezers. 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., 2100
0 × 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 in a 5-fold weight solubilization treatment. 4
C., 60 minutes). This is ultracentrifuged (4 ° C., 11
0000 × g, 90 minutes), and the supernatant was dialyzed (4 ° C., 24 hours) against 0.01 M potassium phosphate buffer (pH 7.0) to obtain an enzyme (α-glucosidase) solution.

(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 mM substrate solution (sucrose dissolved in 0.1 M potassium phosphate buffer, pH 6.3), test substance solution [E-1 and E-2 fractions obtained from 0.1 g of magnolia Was concentrated to dryness, and dissolved in a 50% aqueous dimethyl sulfoxide solution so as to be 1 ml each.] 0.2 ml, and 0.1 ml of the above enzyme solution (1.0 ml in total). This is 3
Reaction at 7 ° C. for 15 minutes, 2M Tris-HCl buffer (pH
7.0) The reaction was stopped using 1.5 ml to prepare a test solution. Next, a coloring reagent [glucose B test Wako (manufactured by Wako Pure Chemical Industries, Ltd.)] 200 μm per well was placed in a 96-well microplate.
After adding 50 μl of the test solution to the mixture and incubating at 37 ° C. for 30 minutes, a microplate reader (BIO RA) was added.
The absorbance at 490 nm was measured with Model D (manufactured by Company D). The absorbance when 0.1 M potassium phosphate buffer (pH 6.3) was added instead of the substrate solution was defined as a blank value, and the value obtained by subtracting this value was defined as _A490s . 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 ₄₉₀ s) / A _490c ] × 100

Example 2 Measurement was carried out in the same manner as in Example 1 except that Koboku was changed to Inchinko.

Example 3 The measurement was carried out in the same manner as in Example 1 except that the magnolia was changed to Chen.

Example 4 Measurement was carried out in the same manner as in Example 1 except that Koboku was changed to Goshuyu.

Example 5 The measurement was performed in the same manner as in Example 1 except that the thick wood was changed to a light load.

Example 6 The same procedure was followed as in Example 1 except that Koboku was changed to Bellflower.

Example 7 The measurement was carried out in the same manner as in Example 1 except that Kobushi was changed to Kiko.

Example 8 The measurement was carried out in the same manner as in Example 1 except that Koboku was changed to Gomiko.

Example 1 (Park), Example 2 (Inchenko),
Fractions of Example 3 (Chen skin), Example 4 (Gusyuyu), Example 5 (Light load), Example 6 (Kikyo), Example 7 (Gikko), Example 8 (Gomiko etc.) Table 1 shows the α-glucosidase inhibitory activity of 1, E-2).

[0026]

[Table 1]

[0027]

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

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A23L 1/307 A23L 1/307 A61K 31/00 603 A61K 31/00 603N C12N 9/99 C12N 9/99 F Terms (reference) 4B018 LB01 LB02 LB05 LB06 LB07 LB08 LB09 LB10 MD66 ME03 MF01 4C088 AB29 AB30 AB38 AB48 AB62 AB65 AC02 BA09 BA10 CA05 CA06 MA52 NA14 ZA70 ZC20 ZC35

Claims (2)

[Claims] 1. An α-glucosidase inhibitor comprising a Chinese herb as an active ingredient. 2. The α-glucosidase inhibitor according to claim 1, wherein an extract obtained by extracting Chinese herbs with water and / or alcohol is used as an active ingredient.