JP3102753B2 - Sugar absorption inhibitor containing saponin mixture derived from acacia shoots and acacia shoots as active ingredients - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention mainly inhibits the absorption of sugar from the stomach and intestine into the body, thereby preventing a rise in blood sugar concentration (blood sugar level) and reducing sugars such as obesity and diabetes. It is related to sugar absorption inhibitors that are effective in protecting the body from physiological disorders caused by overdose, and in particular, Aralia e, a deciduous shrub of Araliaceae.
about glucose absorption inhibitor as an active ingredient saponin mixture extracted terminal buds or shoots or these lata SEEM).

[0002]

BACKGROUND OF THE INVENTION Conventional sugar absorption inhibitors include glycosides represented by gymnema acid (Gymnema leaf component) derived from Gymnema sylvestre in the potato family and digidifin derived from kenponashi in the buckthorn family. Α-Glucosidase (glycolytic enzyme) inhibitors represented by acarbose are already known. However, since the former glycosides tend to exhibit sweetness suppressing activity, it is necessary to use glycosides in order to prevent the occurrence of physiological disorders due to the excessive intake of sugar. It is inherently inappropriate to apply, since the premise of overdose means that it is difficult to control the intake of sugar itself, and in the latter α-glucosidase inhibitor, oligosaccharides such as sucrose On the other hand, the activity of inhibiting absorption of saccharides is remarkable, but the activity of inhibiting absorption of monosaccharides such as glucose is completely absent, so that the range of use is limited and universal practical application is difficult.

[0003]

As described above, with respect to the conventional sugar absorption inhibitors, glycosides are inherently not promising in terms of the sweetness suppressing activity, and α-glucosidase inhibitors are singly used. There is a problem that it cannot be applied to sugars. Therefore, focusing on the problem of sweetness suppressing activity and the problem of being inapplicable to monosaccharides, there is no sweetness suppressing activity, and moreover,
In terms of applicability to monosaccharides, it is possible to provide a sugar absorption inhibitor that meets the clinical or daily demands of diversifying the uses of sugar absorption inhibitors, and to solve the above problems. It is an object of the present invention.

[0004]

In view of the above-mentioned problems of removing sweetness suppressing activity and responding to the demand for diversifying uses of the above-mentioned conventional sugar absorption suppressors, the inventors of the present invention have developed a method in Japan and China. diabetes treatment and medicinal plants cod eaves known to be robustly is valid (Aralia elata SEEM)
Focusing the search target, intensive, as a result of repeated searching and testing evaluation, the saponin mixture as glucose absorption inhibiting component contained in the Aralia elata buds or Aralia elata shoots, of each part of Aralia elata, are often used to date Site (bark,
(Root bark, leaf), and were found to be distinct from the saponin mixture contained therein, and by purifying the same, the sugar absorption inhibitor of the present invention was completed.

[0005]

According to the first aspect of the present invention, Aralia ela
ta SEEM) Top shoots and Aralia elata SEEM shoots
Reverse phase silica gel extract
Water and then methanol
And elute the methanol elution with normal phase silica gel color.
Chloroform-methanol
Elution with a mixed solvent of water and water, the following formula (1)

Embedded image Eratoside G represented by the following formula (2)

Embedded image And the following equation (3)

Embedded image Of saponin mixture containing elatoside I represented by
Eratside G, Eratside H and
Saponin mixture containing elatoside I as active ingredient
This is a method for producing a sugar absorption inhibitor. This saponin mixture has an Rf value of 0.2 to
0.4 Der is, a pale brown powder under dry conditions, water,
Aqueous methanol, aqueous ethanol, methanol, ethanol, dimethyl sulfoxide, soluble in Pyridine, ethyl acetate, chloroform, sparingly soluble der against acetone is, act as an active ingredient of glucose absorption inhibitor. The invention according to claim 2 is the sugar absorption suppression according to claim 1.
In the method for producing the agent, normal phase silica gel column chromatography
By chromatography, chloroform-methanol-water (6:
4: 1) Elatoside G, Eratside H and
For separating saponin mixture containing erythroside and eratoside I
It is.

[0006]

Example (1) Extraction of saponin mixture A mixture of agaric shoot buds (open buds) and agaric shoots (open buds) (3K from Nagano Prefecture)
g) was extracted by heating with 80% aqueous methanol (101) for 5 hours, and the extract was collected by filtration. 80% aqueous methanol (101) was further added to the residue, and the same methanol extraction operation was repeated three times. The extracts were combined, the solvent was distilled off under reduced pressure, and an 80% aqueous methanol extract (160 g) was obtained as a residue. 80% aqueous methanol extract was subjected to reverse phase silica gel column chromatography (C
₁₈ Chromatorex, made by Fuji Silysia Chemical LTD,
1.2 kg) and eluted with water and then methanol. The solvent was distilled off from the eluted portion under reduced pressure with methanol to obtain an extract eluted with methanol (16.0 g). Next, the methanol eluate extract was subjected to normal phase silica gel column chromatography (Silicagel 60, manufactured by Merck, 300 g),
The mixture was eluted with a mixed solvent of chloroform-methanol-water. The solvent was distilled off from the chloroform-methanol-water (6: 4: 1) eluate under reduced pressure to obtain a saponin mixture (9.9 g). When the obtained saponin mixture was developed with chloroform-methanol-water (6: 4: 1) for thin-layer chromatography (Pre-coated Silicagel 0.25 mm manufactured by Merck), the Rf value was 0.2 to 0.4. A spot was observed at position 4. Purification of the saponin mixture obtained here yielded a light brown powdery substance in a dry state. This substance contains water, aqueous methanol, aqueous ethanol, methanol, ethanol, dimetal sulfoxide,
It was found to be soluble in viridine and poorly soluble in ethyl acetate, chloroform and acetone.

(2) Isolation of Elatoside in Saponin Mixture The saponin mixture (1.3 g) was subjected to high performance liquid chromatography [HPLC; equipment: LC-10AS type liquid chromatograph, with C-R6A chromatopack, Shimadzu Manufactured by the factory;
Column: YMC-Pack ODS-A (250 × 20
mmI. D. ) YMC; elution solvent: methanol-1%
Acetic acid aqueous solution (4: 1 V / V); Detector: Reflective index
detector RID-6A, manufactured by Shimadzu Corporation] to obtain a saponin (7 mg) having a chemical structure represented by the following chemical structural formula (1) and referred to as elatoside G. ) And the following chemical structural formula (2)
Having a chemical structure represented by the formula:
ide H) (4 mg) and a saponin (7 mg) having the chemical structure represented by the following chemical structural formula (3) and designated as elatoside I (elatoside I) are isolated and identified. did.

Embedded image

Embedded image

Embedded image

(3) Dosage form The saponin mixture described in the above (1) is ingested in the form of an oral dosage form or a parenteral dosage form for use in treatment, prevention and health. Oral dosage forms include powders, tablets, emulsions, capsules, teas, granules, and liquids (including tinctures, fluid extracts, alcoholic beverages, suspensions, limonades) in the usual manner. Can be The dosage in the case of an oral dosage form varies depending on the purpose of treatment, prevention, health and the like, but is 10 mg to 100 m / day for an adult.
g, preferably 50 mg to 150 mg of the saponin mixture may be administered in two or three divided doses. In addition, foods for specified health use, JSD foods, special nutritional foods, dietary supplements,
It can also be added as a food additive to health foods and the like. Examples of parenteral dosage forms include ointments, plasters, liquids (including spirits, tinctures, and lotions), poultices (happs, pastas), coatings, sprays, dusting agents, A liniment (rubbing agent), a cream, an emulsion, and a bath are exemplified as ordinary methods. As for the dosage in the case of a parenteral dosage form, it is sufficient to mix the saponin mixture at a concentration of 0.01 to 5%, preferably 0.1 to 1%.

(4) Evaluation of sugar absorption inhibitory function (blood sugar level increase inhibitory effect) of saponin mixture Male wistar rats (body weight 1) fasted for 20 to 24 hours
25 to 155 g), orally administered a test drug obtained by dissolving the saponin mixture according to the above (1) in water (5 ml / kg), and after 30 minutes, glucose (0.5 g / kg) was orally administered. Was administered. Thereafter, under anesthesia restraint at the time of blood collection, blood collected from the jugular vein about 0.5 ml each time was cooled with ice water, and serum was separated from the blood by centrifugation,
The glucose concentration in the separated serum was measured by the glucose oxidase method according to a conventional method, and the measurement results as shown in Table 1 were obtained.

[Table 1] Suppressive action on blood glucose level in glucose-loaded rats (saponin mixture) Treatment group Body weight 1kg No. of dose per blood glucose (mg / dl) (Mg / kg) 0.5hr 1.0hr 2.0hr Normal group 5 68.4 ± 3.4 95.6 ± 4.3 91.4 ± 3.2 Control group 8 146.0 ± 5.3 130.3 ± 6.8 102.4 ± 1.5 Glucose load 77.6 ± 5.3 34.7 ± 6.8 11.0 ± 1.5 Seed buds, 200 5 95.5 ± 6.5 117.3 ± 9.1 111.7 ± 8.9 Shoot saponin mixture In Table 1, the blood glucose level in the “normal group” column is the blood glucose level of the rat group to which glucose was not administered orally, and the blood glucose level in the “control group” column is that of glucose administered orally. It is a blood glucose level of a rat group. And the "glucose load"
The blood glucose level in the column indicates an increase in the blood glucose level in the “control group” column with respect to the blood glucose level in the “normal group” column. On the other hand, in Table 1, the blood glucose level in the column of “mixture of acrophora shoots and shoot saponins” is obtained by orally administering the saponin mixture according to the above (1) extracted from a mixture of acrophora shoots and shoots of shoots. It shows the increase in blood glucose level when glucose was orally administered under the same conditions as in the case of the “control group” column. Therefore, the blood sugar level increase inhibitory effect is evaluated based on the decrease in the blood sugar level in the column of “mixture of shoot aerial shoot and shoot saponin” in comparison with the blood sugar level in the column of “control group”. The elatoside G. according to the above (2) isolated from the saponin mixture according to the above (1).
H. Table 2 shows the results of evaluation of the blood sugar level increase inhibitory effect of each of I by the same method as in Table 1 above.

[Table 2] Inhibitory effect of blood glucose level increase in glucose-loaded rats (Eratocide GHI) Treatment group Body weight 1kg No. of dose per blood glucose (mg / dl) (Mg / kg) 0.5hr 1.0hr 2.0hr Normal group 5 68.4 ± 3.4 95.6 ± 4.3 91.4 ± 3.2 Control group 8 146.0 ± 5.3 130.3 ± 6.8 102.4 ± 1.5 Glucose load 77.6 ± 5.3 34.7 ± 6.8 11.0 ± 1.5 Seed buds, 200 5 95.5 ± 6.5 117.3 ± 9.1 111.7 ± 8.9 Shoot saponin mixture elatoside G 100 4 97.8 ± 10.1 114.0 ± 8.6 111.0 ± 9.3 elatoside H 100 3 89.3 ± 4.3 129.7 ± 7.9 124.0 ± 21.1 elatoside I 100 5 86.2 ± 6.7 118.2 ± 5.8 113.8 ± 6.9

[0010]

According to the present invention, there is provided a Aralia elata buds or cod <br/> eaves shoots or we extracted saponins mixture of Aralia elata, bark is known for use as medicinal plants, roots A saponin mixture having a Rf value of 0.2 to 0.4 determined by thin-layer chromatography, which is obtained by purifying a saponin mixture different from the saponin mixture extracted from a skin or the like, as an active ingredient, is contained in the sugar absorption inhibitor. With this configuration, the saponin mixture suppresses the absorption of saccharides in the stomach and intestine of an organism and suppresses an increase in blood sugar concentration (blood sugar level). In addition, since the saponin mixture lacks sweetness suppressing activity, when added to foods and drinks, there is no reduction in sweetness, and not only oligosaccharides such as sucrose, but also monosaccharides such as glucose. Since exhibits glucose absorption inhibiting action also, excellent effect that diversification of applications can be achieved are obtained. The sugar absorption inhibitor of the present invention, in view of the above effects,
It has great industrial value as a food additive to food and drink for obese and diabetic patients.

Continued on the front page (58) Fields surveyed (Int. Cl. ⁷ , DB name) A61K 35/78 A61K 31/70 C07H 15/256 CA (STN) REGISTRY (STN)

Claims (2)

(57) [Claims] 1. An extract from a mixture of agar buds (Aralia elata SEEM) apical buds and a cotyledon (Aralia elata SEEM) shoot eluted with methanol is subjected to reversed-phase silica gel column chromatography and eluted with water and then methanol. The methanol eluted portion was subjected to normal phase silica gel column chromatography and eluted with a mixed solvent of chloroform / methanol / water to obtain the following formula (1). Eratoside G represented by the following formula (2): And the following formula (3): A method for producing a sugar absorption inhibitor comprising a saponin mixture containing eratoside G, eratoside H and eratoside I as an active ingredient, comprising a step of separating a saponin mixture containing eratoside I represented by the formula: 2. The sugar absorption according to claim 1, wherein the saponin mixture containing eratoside G, eratoside H and eratoside I is separated from the eluate of chloroform-methanol-water (6: 4: 1) by normal phase silica gel column chromatography. Method for producing inhibitors.