JP2000239170A - Diabetes meritus-treating agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject new treating agent having less limitation in its use, and having a suitable symptom-moderating effect by containing sesaminol triglucoside. SOLUTION: This treating agent contains sesaminol triglucoside of the formula sesaminol 2'-O-β-D-glucopyranosyl(1→2)-O-[β-D-glucopyranosyl(1→6)]-β-D- glucopyranoside}. The compound of the formula exists in seeds of sesame in the natural world, and is obtained e.g. by defatting and crushing the sesame seeds, extract-treating the crushed material with 80% ethanol-water, fractionating the extract by using a silica gel column, and further subjecting the fractionated material to a liquid chromatography using an ODS column for fractionally collecting. As to the method for administration, it can be administered orally, and the daily dosage is usually several hundred to 20,000 mg as the sesminol glucoside for an adult.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a therapeutic agent for diabetes. At present, the number of diabetics is steadily increasing, and the Ministry of Health and Welfare has found that including the reserve army, it will reach 10% of the Japanese population. Diabetes is a disease that causes symptoms such as hyperglycemia due to lack of insulin action, but it induces serious complications that are life-threatening, and therefore has a great social interest in its treatment. The present invention relates to such a therapeutic agent for diabetes, and particularly to a therapeutic agent for diabetes containing sesameol triglucoside derived from sesame seed as an active ingredient.

[0002]

2. Description of the Related Art Conventionally, insulin is generally used as a therapeutic agent for diabetes. However, insulin has a problem in that its administration method, dosage and timing of administration are greatly restricted, and in some cases, hypoglycemia is caused and there is even a risk of death. For this reason,
There is a demand for a therapeutic agent having a blood sugar lowering effect that has less restrictions on its use and does not lower blood sugar more than necessary.

[0003]

The problem to be solved by the present invention is to provide a novel therapeutic agent for diabetes, which is less restricted in its use and has an appropriate symptom-relieving action.

[0004]

Means for Solving the Problems Thus, the present inventors have
As a result of researching to solve the above-mentioned problems, it has been found that those containing a water-soluble sesaminol triglucoside having a specific structure contained in sesame seed as an active ingredient are correct and suitable.

[0005] That is, the present invention relates to a therapeutic agent for diabetes comprising sesaminol triglucoside represented by the following formula 1 as an active ingredient.

[0006]

(Equation 1)

In the present invention, sesaminol triglucoside represented by the formula 1 is sesaminol 2′-O-β-D
-Glucopyranosyl (1 → 2) -O- [β-D-glucopyranosyl (1 → 6) -β-D-glucopyranoside, which is naturally present in sesame seeds. The present invention does not particularly limit the method for separating the sesaminol triglucoside represented by the formula 1 from sesame seeds. Examples of the separation method include a method disclosed by Katsuzaki et al. {Human Chemistry (phytochemistry) Vol. 773-776,
1994｝ is applicable. That is, the sesame seeds were defatted and crushed, extracted with 80% ethanol water, and the extract was fractionated using a silica gel column, and the fraction was further subjected to liquid chromatography using an ODS column. The sesaminol triglucoside represented by the formula 1 is fractionated as a highly purified product.

[0008] The sesaminol triglucoside represented by the formula 1 has an appropriate symptom-relieving action in the treatment of diabetes, as will be described later in detail. Therefore, as the active ingredient of the therapeutic agent for diabetes according to the present invention, sesaminol triglucoside itself isolated from sesame seeds can be used, and any of those containing such sesaminol triglucoside as described below. Separated products and defatted products can also be used.

First, the separated product will be described. Separation-treated product used as an active ingredient of the therapeutic agent for diabetes according to the present invention is obtained by defatting sesame seeds and separating the defatted product from water, an aqueous extract, or an extract obtained by performing an extraction treatment using a mixed solvent thereof. Is obtained. Such separated products include 1) those obtained by removing the solvent from the above-mentioned extract, and 2) treatment of the above-mentioned extract with an adsorbent, thereby adsorbing solvent-soluble impurities, and thus removing impurities. A product obtained by removing the solvent from an extract in which sesaminol triglucoside is dissolved after adsorption and removal, 3)
The extract obtained by treating the above-mentioned extract with an adsorbent, adsorbing the adsorbable component containing sesaminol triglucoside thereon, and then desorbing and eluting the adsorbable component from the adsorbent is included.

In the above 1) to 3), the water-soluble solvent for obtaining the extract is preferably ethanol, and the mixed solvent is preferably a water-ethanol mixed solvent.
Among these separated products 1) to 3), 3) is preferred, and among them, those obtained through the following first, second and third steps are more preferred. 1st step: Sesame seeds are defatted, and 50%
A step of extracting a hot water-soluble component using hot water at a temperature of at least 100 ° C. to obtain an extract in which the hot water-soluble component is dissolved. Second step: the extract obtained in the first step and a vinyl aromatic hydrocarbon-based polymer The polymer-based adsorbent made by contacting with the polymer adsorbent made of coalescing to adsorb the adsorptive component contained in the hot water-soluble component in the extract to the polymer adsorbent. Step of obtaining an agent Third step: 40 to 8 from the polymer adsorbent obtained in the second step
A step of desorbing and eluting sesaminol triglucoside contained in the adsorptive component adsorbed on the polymer-based adsorbent using 0% by volume of ethanol water to obtain ethanol water in which the sesaminol triglucoside is dissolved

The separated product is obtained by removing the solvent from the aqueous ethanol solution containing sesaminol triglucoside obtained in the third step by an appropriate method.

In the first step, the temperature of the hot water used for extraction is set to 50 ° C. or higher, preferably 60 ° C. or higher. The ratio of hot water to degreased product is 10
It is preferable to set the hot water to 700 to 1000 parts by weight per 0 parts by weight. The extract obtained by the extraction treatment has the hot water-soluble component contained in the defatted product dissolved therein.

In the second step, the extract obtained in the first step is brought into contact with a polymer adsorbent made of a vinyl aromatic hydrocarbon-based polymer to remove the adsorptive components contained in the extract to the high degree. This is a step of adsorbing on a molecular adsorbent. As the polymer-based adsorbent made of a vinyl aromatic hydrocarbon-based polymer used here, a polymer-based adsorbent made of a styrene-divinylbenzene copolymer is preferable, and among them, a BET surface area of 80 to 1000 is preferable.
m ² / g and a porosity (helium porosity, the same applies hereinafter) of 0.1 to 1.5 ml / g are preferred. The proportion of the polymer adsorbent used is preferably 1/10 to 1/30 times that of the extract.

[0014] In the second step, it is preferable to add the polymer-based adsorbent to the extract and stir at a temperature of 30 to 70 ° C. Thus, by bringing the extract into contact with the polymer-based adsorbent, the adsorptive component contained in the hot water-soluble component in the extract is adsorbed by the polymer-based adsorbent. The system adsorbent is removed by a known solid-liquid separation method.

In the third step, sesaminol contained in the adsorptive component adsorbed on the polymer adsorbent from the polymer adsorbent obtained in the second step using 40 to 80% by volume of ethanol water is used. This is a step of desorbing and eluting triglucoside.
The ethanol water used for desorption and elution is preferably 45 to 75% by volume of ethanol water, and the temperature for desorbing and eluting sesaminol triglucoside is preferably 15 to 50 ° C. Is preferably 2 to 4 times the amount of the polymer-based adsorbent. As a method for desorbing and eluting sesaminol triglucoside from a polymer adsorbent, 1) a method in which ethanol water is passed through a column filled with a polymer adsorbent having adsorbable components adsorbed thereto under normal pressure or pressure, 2) A method in which a high molecular weight adsorbent to which an adsorbent component is adsorbed is added to ethanol water and stirred. When sesaminol triglucoside is desorbed and eluted by the method 2), the polymer adsorbent after desorption and elution is separated by a known method such as decantation, centrifugation, or filtration.

When sesaminol triglucoside is desorbed and eluted from the polymer-based adsorbent, ethanol water in which sesaminol triglucoside is dissolved is obtained. The ethanol and water are removed from the ethanol water to obtain a separated product. This separated product contains 20 to 30% by weight of sesaminol triglucoside on a dry matter basis.

In the case where the sesameol triglucoside represented by the formula 1 is separated from the sesame seed, and in the case where the above-mentioned separated product is obtained from the sesame seed, the sesame seed that has not been molted or that has been molted is used as the sesame seed. Although sesame seeds can be used, it is preferable to use dehulled sesame seeds in order to facilitate subsequent degreasing treatment and further extraction treatment, and it is more preferable to use crushed sesame seeds in addition to dehulling.
It is preferable to use unroasted sesame seeds. Such sesame seed degreasing treatment includes a method of pressing and squeezing oil and a method of extracting with a solvent. When degreased by pressing and squeezing, the defatted product is obtained as a pressed oil residue, and when degreased by solvent extraction, the defatted product is obtained as an extraction residue. As a solvent for the degreasing treatment by solvent extraction, an aliphatic hydrocarbon such as hexane or an aliphatic lower alcohol such as ethanol can be used.

Finally, a defatted product used as an active ingredient of the therapeutic agent for diabetes according to the present invention will be described. This defatted product is a defatted product obtained by subjecting sesame seeds to a degreasing treatment by solvent extraction. In this case as well, sesame seeds that have not been molted or hulls that have been molted can be used as sesame seeds.However, in order to facilitate subsequent degreasing treatment, it is preferable to use molted sesame seeds. In addition, it is more preferable to use a crushed one. It is preferable to use unroasted sesame seeds. Specifically, for example, after adding and stirring an aliphatic hydrocarbon such as hexane in an amount of 5 to 10 times as an extraction solvent to sesame seeds, a solid content is taken out from the extraction system, and the extraction solvent is distilled off from the taken out solid content. do it,
A defatted product is obtained as an extraction residue.

Among the above defatted products, the defatted product used as an active ingredient of the antidiabetic agent according to the present invention includes sesaminol triglucoside in terms of dry matter of 0.3%.
Those containing oil at a ratio of not less than 10% by weight and not more than 10% by weight are preferred, and those containing oil at a ratio of not more than 3% by weight are particularly preferred. This is because as a result, the intake of lipids that is inconvenient for diabetics is suppressed. Such a defatted product can be obtained through the following first, second and third steps. First step: After adding an aliphatic hydrocarbon as an extraction solvent to the dehulled sesame seeds and stirring, a solution component is removed from the extraction system to obtain a solid content containing the extraction solvent at a ratio of 10% by weight or less. Step 2: Step of adding water to the solid content to obtain a hydrated substance Third step: Step of removing the extraction solvent from the hydrated substance to obtain a defatted product The first step, the second step, and the third step as described above. The defatted product obtained through the process is known per se, and is disclosed in JP-A-9-987.
No. 50 publication.

The therapeutic agent for diabetes according to the present invention, as described above, comprises 1) sesaminol triglucoside as an active ingredient, 2) a separated product as an active ingredient, or 3) defatting. A substance is used as an active ingredient. These can be administered orally, and the dose is usually several times a day as sesaminol triglucoside per adult.
The range is from 00 to 2000 mg. The therapeutic agent for diabetes according to the present invention containing these as active ingredients has an effect of appropriately lowering blood sugar level and urinary sugar for adults exhibiting diabetic symptoms.

[0021]

DESCRIPTION OF THE PREFERRED EMBODIMENTS Preferred embodiments of the present invention include the following 1) to 3). 1) Sesame seeds are crushed, hexane is added to the crushed material, and an extraction treatment is performed. A solid content is taken out from the extraction system, and the taken out solid content is dried to obtain a defatted product. 80% by weight of ethanol water is added to the defatted product for extraction treatment, the extract is taken out of the extraction system, and the taken out extract is dried to obtain an extract. 80% by weight of ethanol water was added to this extract, and the solution was adsorbed on a polystyrene gel column.
This is concentrated. The concentrate was subjected to liquid chromatography using an ODS column, and methanol / water = 1/1
Using (V / V) as a mobile phase, a fraction flowing out at a retention time of 15 to 17 minutes is collected, and the collected solution is dried to obtain a substantially pure form of sesaminol triglucoside (S-1). . A therapeutic agent for diabetes containing the sesaminol triglucoside (S-1) as an active ingredient.

2) A defatted product is obtained in the same manner as in 1) above. Hot water at 100 ° C is added to this defatted product for extraction,
Extract the extract from the extraction system. A dry granular polymer adsorbent made of a styrene-divinylbenzene copolymer (BET surface area: 300 m ² / g, porosity: 0.
(70 ml / g), and after stirring, the solution is removed to obtain a wet polymer adsorbent. A glass column is filled with the wet polymer adsorbent, and 60% by volume ethanol water is passed through the glass column to obtain an eluate. The eluate was dried, and the separated product containing 27.0% by weight of sesaminol triglucoside (S-
Obtain 2). An antidiabetic agent comprising the separated product (S-2) as an active ingredient.

4) Roll-pressing the molted sesame seeds,
Hexane is added thereto as an extraction solvent to perform an extraction treatment, and a solid content is extracted from the extraction system. The solids taken out are sprayed with water to make them hydrated. The extraction solvent was distilled off from the hydrated product under reduced pressure to reduce sesaminol triglucoside to 0.43%.
A defatted product (S-3) containing at a weight percentage is obtained. A therapeutic agent for diabetes containing the defatted product (S-3) as an active ingredient.

Hereinafter, the structure and effects of the present invention will be described in detail with reference to examples, but the present invention is not limited to these examples. In the following examples,
Unless indicated otherwise, parts means parts by weight and% means% by weight.

[0025]

Example 1 Test Category 1 Preparation of Sesaminol Triglucoside (S-1) Chinese sesame seeds (250 g) were ground and placed in a flask. One liter of hexane was added, the mixture was stirred at room temperature for 5 hours, and then filtered. The same operation was further repeated twice on the solid content after filtration, and the solid content was dried by ventilation at room temperature to obtain 115 g of a defatted product. 115 g of the defatted product was placed in a flask, 736 g of ethanol and 184 g of water were added, and the mixture was stirred at room temperature for 15 hours, followed by filtration to obtain 355 g of a filtrate. The filtrate was concentrated under reduced pressure at a temperature of 40 ° C. or less to 80 g, and further lyophilized to obtain 2.7 g of an extract. Dissolve 0.5 g of this extract in 80% ethanol aqueous solution,
Polystyrene gel (trade name Amberlite XAD-2,
(Rohm and Haas Co., Ltd.) from the top of the glass column and adsorbed to polystyrene gel. As the mobile phase, starting with 75 ml of water, 75 ml of a 20% aqueous methanol solution, 75 ml of a 40% aqueous methanol solution, 6 ml
A 0% aqueous methanol solution (75 ml), an 80% aqueous methanol solution (75 ml) and methanol (75 ml) were successively flowed, and the eluate eluted with the 60% aqueous methanol solution was collected and concentrated. This concentrate was further fractionated using liquid chromatography under the following conditions.

Stationary phase: Develosil ODS-5 (trade name, manufactured by Nomura Chemical Co., Ltd.) Column diameter: 10 mm Column length: 250 mm Mobile phase: methanol / water = 1/1 (v / v) Mobile phase flow rate: 2.5 ml / Min Detection: UV 280 nm The fraction which elutes at a retention time of 15 to 17 minutes is collected, concentrated under reduced pressure at a temperature of 40 ° C. or less, and further freeze-dried to give sesaminol triglucoside in a substantially pure form. (S-1) 19.0 mg was obtained. This indicates that this is a sesaminol triglucoside represented by the formula 1, that is, sesaminol 2′-O-β-D-glucopyranosyl (1 → 2) -O- [β-D-glucopyranosyl (1 →
6)]-β-D-glucopyranoside was determined by the following various analysis results.

Analysis result Mass spectrum [M−H] ⁻ ; m / z 855 UV absorption spectrum λmax (log ε); 290 nm (3.82), 23
6 nm (3.80) Specific rotation [α] _D ; -9.75

¹ H-NMR δ chemical shift (attributed hydrogen) 2.88 (H-1), 4.65 (H-2), 3.76
(H-4a), 4.16 (H-4e), 2.88 (H-
5), 5.16 (H-6), 3.98 (H-8a),
4.16 (H-8e), 6.91 (H-3 '), 6.7
5 (H-6 ′), 6.75 (H-2 ″), 6.77 (H
-5 "), 6.75 (H-6"), 5.87 (-OC
H ₂ —O—), 5.91 (—O—CH ₂ —O—), 5.1
9 (G1), 3.84 (G2), 3.81 (G3),
3.50 (G4), 3.72 (G5), 3.80 (G6
a), 4.09 (G6b), 4.85 (G1 ′),
29 (G2 ') 3.48 (G3'), 3.40 (G
4 '), 3.30 (G5'), 3.58 (G6a '),
3.65 (G6b '), 4.39 (G1 "), 3.21
(G2 "), 3.39 (G3"), 3.29 (G
4 "), 3.17 (G5"), 3.60 (G6a "),
3.62 (G6b ")

¹³ C-NMR δ chemical shift (attributed carbon) 55.4 (C-1), 83.0 (C-2), 73.5
(C-4), 55.4 (C-5), 87.4 (C-
6), 74.5 (C-8), 125.4 (C-1 ′),
150.0 (C-2 '), 100.4 (C-3'), 1
49.4 (C-4 ') 144.8 (C-5'), 10
7.5 (C-6 '), 136.3 (C-1 "), 10
8.8 (C-2 "), 149.6 (C-3"), 14
9.0 (C-4 "), 110.5 (C-5"), 12
2.0 (C-6 "), 103.4 (-O-CH 2 -O
-), 103.4 (-O-CH 2 -O -), 101.1
(G1), 82.4 (G2), 77.7 (G3), 7
1.4 (G4), 77.4 (G5), 70.5 (G
6), 103.9 (G1 ′), 76.0 (G2 ′), 7
7.7 (G3 '), 71.4 (G4'), 78.0 (G
5 '), 63.0 (G6'), 104.7 (G1 "),
75.3 (G2 "), 77.9 (G3"), 71.8
(G4 "), 78.1 (G5"), 62.4 (G6 ")

Test Category 2 Preparation of Separated Product (S-2) Sesame seeds were pressed and oiled, and 900 parts of hot water at 100 ° C. was added to 100 parts of defatted material obtained as the pressed oil residue.
After stirring for minutes, the mixture was filtered under reduced pressure to obtain 700 parts of an extract. A BET surface area of 300 m ² / g was added to 700 parts of this extract.
After adding 35 parts of a polymer-based adsorbent made of a styrene-divinylbenzene copolymer having a porosity of 0.70 ml / g, stirring the mixture at 60 ° C. for 3 hours, and then decanting the polymer-based adsorbent in a wet state. 40 parts of the agent were obtained. A glass column was filled with 40 parts of the wet polymeric adsorbent, and 100 parts of 60% by volume ethanol water was passed through the column at 20 ° C. to obtain 100 parts of an eluate. The eluate was concentrated and freeze-dried to reduce sesaminol triglucoside to 27.0%.
(S-2) was obtained. The content of sesaminol triglucoside in the separated product (S-2) was determined by dissolving the separated product in 60% by volume ethanol water and using a high performance liquid chromatography method. As a standard sample at this time, sesaminol triglucoside obtained in Test Category 1 was used.

Test Category 3-Preparation of defatted product (S-3) 1 kg of commercially available dehulled sesame seeds was crushed by a roll pressing machine, taken into a container, and 1 liter of hexane was added.
After stirring at 0 ° C. for 1 hour, the contents were separated into a solid content and a solution content by a suction filter at room temperature. The extraction solvent content of the solid was 1.5%. 80 g of water was sprayed on 200 g of this solid content, and stirred to obtain a water-containing substance. This hydrate 2
80 g was transferred to a vacuum dryer, and 80 ° C. under a reduced pressure of 1 KPa.
Then, the extraction solvent was distilled off over 5 hours to obtain a defatted product (S-3) containing 0.43% of sesaminol triglucoside and 1.5% of oil. The defatted product (S-3) did not substantially contain hexane used as an extraction solvent. The content of sesaminol triglucoside, oil and hexane in the defatted product (S-3) is described in JP-A-9-9875.
It was determined according to the method described in Japanese Patent Publication No.

Test Category 4 (Evaluation 1) Evaluation of Serum Sugar Concentration Breeding of Experimental Animals Wistar rats (4 weeks old, 200 g body weight) were divided into 5 groups A to E, each group consisting of 8 rats. . 4 of B, C, D, E
Streptozotocin was administered to rats in the group at 6 / kg body weight.
It was administered intraperitoneally at a rate of 0 mg to cause diabetes. Thereafter, each group was fed with a feeding diet having a composition as shown in Table 1 at a rate of 10 g / day, and reared for 4 weeks.

[0033]

[Table 1]

In Table 1, a normal diet: a diet for rats consisting of 25% casein, 3.5% mineral mixture, 1% vitamin mixture, 5% corn, 4% cellulose and 61.5% corn starch.

Evaluation of sugar concentration in serum After breeding for 4 weeks, blood was collected from experimental animals, and serum was separated according to a standard method. The sugar concentration in the obtained serum was measured using a sugar concentration measurement kit (trade name: Glucose-Test Wako, manufactured by Wako Pure Chemical Industries, Ltd .; the same applies hereinafter). Table 2 shows the measurement results.
It was shown to.

[0036]

[Table 2]

In Table 2, P: risk factor for group B

Test Category 5 (Evaluation 2) ・ Evaluation of urine sugar concentration ・ ·
Urine was collected 4 weeks after the start of breeding. The sugar concentration of the collected urine was measured with a commercially available kit for measuring sugar concentration. Table 3 shows the measurement results.

[0039]

[Table 3]

In Table 3, P: risk factor for group B

[0041]

As is clear from the above, the present invention described above has an effect of having an appropriate symptom-relieving action in the treatment of diabetes.

 ──────────────────────────────────────────────────続 き Continuing on the front page (72) Koji Uchida Inventor Koji Uchida 3-252 Shinonokaze 3-chome, Midori-ku, Nagoya-shi, Aichi 1-32 Waterfall Water House 1-372 (72) Inventor Michitaka Naito 47, Takikawacho, Showa-ku, Nagoya-shi, Aichi Prefecture Address No. 5 Mitsui Norin Yachigo Heights 817 (72) Inventor Akashi Kiang 349 Miyatocho, Showa-ku, Nagoya-shi, Aichi 1 Friendship C room F-term (reference) 4C057 BB04 DD01 KK02 4C086 AA01 AA02 EA04 GA17 MA01 MA04 MA52 NA14 ZC35 4C088 AB12 AC04 BA09 BA13 BA23 BA32 CA05 CA13 MA52 NA14 ZC35 ZC54

Claims (7)

[Claims] 1. An antidiabetic agent comprising sesaminol triglucoside represented by the following formula 1 as an active ingredient. (Equation 1) 2. A separated product obtained by subjecting sesame seeds to a defatting treatment and subjecting the defatted product to a separation treatment from an extract obtained by performing an extraction treatment using water, a water-soluble solvent or a mixed solvent thereof is used as an active ingredient. Diabetes treatment. 3. The therapeutic agent for diabetes according to claim 2, wherein the separated product is obtained through the following first step, second step and third step. 1st step: Sesame seeds are defatted, and 50%
A step of extracting a hot water-soluble component using hot water at a temperature of at least 100 ° C. to obtain an extract in which the hot water-soluble component is dissolved. Second step: the extract obtained in the first step and a vinyl aromatic hydrocarbon-based polymer The polymer-based adsorbent made by contacting with the polymer adsorbent made of coalescing to adsorb the adsorptive component contained in the hot water-soluble component in the extract to the polymer adsorbent. Step of obtaining an agent Third step: 40 to 8 from the polymer adsorbent obtained in the second step
A step of desorbing and eluting sesaminol triglucoside contained in the adsorptive component adsorbed on the polymer-based adsorbent using 0% by volume of ethanol water to obtain ethanol water in which the sesaminol triglucoside is dissolved 4. An antidiabetic agent comprising, as an active ingredient, a defatted product obtained by defatting sesame seed by solvent extraction. 5. The defatted product is obtained by defatting dehulled sesame seeds, and contains sesaminol triglucoside in an amount of 0.3% by weight or more and an oil content of 10% by weight or less on a dry matter basis. Item 4. The therapeutic agent for diabetes according to Item 4. 6. The therapeutic agent for diabetes according to claim 5, wherein the defatted product contains an oil content of 3% by weight or less. 7. The method according to claim 1, which is an oral administration preparation.
7. The therapeutic agent for diabetes according to 4, 5 or 6.