JP4727790B2 - Anti-obesity agent - Google Patents

Description

【００１９】
表１に示したように、カシュウのエタノール抽出物は、１２５μｇ／ｍｌの反応液濃度でα―アミラーゼ活性を完全に阻害するとともに、その阻害度合は抽出液濃度に依存することが明らかとなった。
【００２０】
【実施例２】
（水抽出）
乾燥カシュウ５００ｇを粉砕し、この粉末に２５００ｍｌの精製水を加え、９５〜１００°Ｃまで昇温させて２０分間抽出した。得られた抽出液を濾過後、減圧濃縮し、さらに凍結乾燥の方法によって乾燥させて目的のα―アミラーゼ阻害剤の抽出物５３．６ｇを得た。
【００２１】
（５０％阻害活性濃度の算出）
実施例１、２で得られたカシュウ抽出物のα−アミラーゼ阻害活性率を検討し、５０%阻害活性濃度（ＩＣ５０：μｇ・乾燥重量／ｍｌ・反応液）を算出した。なお、小麦種子由来のα−アミラーゼ阻害物質（ＳＩＧＭＡ社製、Ｔｙｐｅ１）を指標阻害剤としてそのＩＣ５０値を算出し比較した。それぞれの結果を表２に示した。
【００２２】
【表２】

【００２３】
表２に示されるように、特に、本発明のカシュウの含水エタノール抽出物を有効成分とするα−アミラーゼ阻害物質は、指標阻害剤の小麦種子由来物質と比較して６倍ほど高い阻害活性を示し、これまで報告されている天然物由来の阻害物質素材と比較しても十分に強い作用を有するものであることが明らかとなった。
【００２４】
【実施例３】
（製剤）
実施例１で得られたカシュウ抽出物を用い、以下の成分組成で調製した。
カシュウ抽出物 １０重量％
乳糖 ７８重量％
結晶セルロース ８重量％
カルボキシメチルセルロースカルシウム ２重量％
ステアリング酸マグネシウム ２重量％
上記の材料を均一に混合し、精製水で混練りした後、乾燥させて単発式打錠機にて錠剤を製造した。
【００２５】
【実施例４】
（顆粒剤）
実施例３（錠剤）で得た錠剤を粉砕、整粒、篩別して１０〜３０メッシュの顆粒剤を得た。
【００２６】
【発明の効果】
本発明によれば、カシュウからの溶媒抽出物を主成分とするα−アミラーゼ阻害物質が提供される。本発明によって得られるα−アミラーゼ阻害物質は、従来のα−アミラーゼ阻害剤と比較してα−アミラーゼ阻害が高く、しかも低濃度で有効であり、さらに植物天然物由来で安全性に優れている。また、本発明のα−アミラーゼ阻害物質は、食品中に含有することで,積極的な食餌療法により、肥満症の予防および改善のために使用することができる。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a saccharide-degrading enzyme inhibitor comprising, as an active ingredient, an extract from kashuu (scientific name: Polygonum multiflorum), and more specifically, suppresses digestion and absorption of a saccharide that serves as an energy source in the body. The present invention relates to an α-amylase inhibitor, an anti-obesity agent, and a food for treating and preventing obesity by inhibiting.
[0002]
[Prior art]
α-Amylase is known as one of digestive enzymes secreted from spleen and salivary glands of animals, and hydrolyzes α-1,4 bonds of polysaccharides such as starch, amylose, and amylopectin.
Further, there is a method of inhibiting the activity of α-amylase and suppressing digestion and absorption of nutrients from the intestinal tract by using an α-amylase inhibitor. This makes it possible to prevent and treat obesity. That is, the obesity-preventing agent and food produced by containing an α-amylase inhibitor are suitable for the treatment of overweight and obese patients, and can also be applied as disease prevention for healthy people.
[0003]
Since long ago, research and development of α-amylase inhibitors have been carried out. Among them, in particular, oligosaccharides produced by microorganisms and peptide amylase inhibitors have been reported and are commercially available as pharmaceuticals. Further, as an example, for plants derived from plants, bay (JP-A-4-27389), guava (JP-A-7-59539), Keihi (JP-A-9-27579), maou (JP-A-9-2963), Extracts from Buttonpi (Japanese Patent Laid-Open No. 9-2966), Oyster (Japanese Patent Laid-Open No. 10-158180), etc. are disclosed as anti-obesity materials.
[0004]
[Problems to be solved by the invention]
As described above, many α-amylase inhibitors have been developed so far. However, plant-derived materials have not been sufficiently put into practical use for pharmaceuticals and foods. Therefore, an object of the present invention is to provide a plant-derived naturally-occurring α-amylase inhibitor that can prevent and improve obesity by daily use, and is effective and safe.
[0005]
[Means for Solving the Problems]
The present inventor has conducted extensive research by screening novel α-amylase inhibitors that improve obesity symptoms from spices, herbs, herbal medicines, etc. The inventors have found an α-amylase inhibitor that is excellent in effectiveness and useful for the prevention and improvement of obesity, and completed the present invention.
[0006]
The present invention comprises the following items.
(Item 1) An anti-obesity agent containing a dry ethanol (50% ethanol) extract of dried oysters as an active ingredient.
[0007]
DETAILED DESCRIPTION OF THE INVENTION
Kashuu (Japanese name: Tsurudokudami) is a perennial vine that belongs to the family Tadeceae, and its origin is in China, but it has come to Kyoho year and grows in various parts of the country. The dried tuberous root is called “Noritori”, and Kampo has effects such as prosthesis, blood replacement, and tonic. Roots contain oxymethylanthraquinones such as emodin, chrysophanol, and fistion, and have been reported to lower blood cholesterol, lower blood pressure, antibacterial, and antioxidant. However, there is no report of an α-amylase activity inhibitor as an anti-obesity action.
[0008]
The collected cashew may be extracted in an undried state, but is usually extracted after drying. In addition, dried shushu generally marketed as a Chinese medicine can also be used. Their properties may be in the form of chips or powders. These α-amylase-inhibiting active ingredients can be obtained by immersing in water or a hydrophilic organic solvent. As an extraction method, heat extraction or continuous extraction may be performed. The obtained extract can be dried by freeze drying, spray drying, or the like.
[0009]
The α-amylase-inhibiting substance of the present invention is an extract containing the above-described extract from oysters as an active ingredient. When used as a drug, for example, diluted with a liquid carrier such as water, ethylene glycol, ethanol, polyethylene glycol, or a non-toxic carrier such as lactose, starch, cellulose, and the like, tablets, granules, Soft capsules, pills, syrups, etc.
[0010]
Since the α-amylase inhibitor of the present invention has an anti-obesity action, for example, it is added to liquid foods such as soft drinks and nutraceutical drinks, and further to solid foods such as crackers, snacks, cookies, and candy. It can be used as a dietary diet for patients or as a disease-preventing diet for healthy people. The addition amount at this time is particularly preferably 0.01 to 2% by weight. Furthermore, the preparation containing the α-amylase inhibitor of the present invention can suppress or delay the rapid absorption of saccharides after eating into the body by taking it before, during or after a meal. .
[0011]
The volume used is 2 to 200 mg / kg body weight, preferably 1 to 150 mg / kg body weight per day for the extract, and is usually taken 1 to 4 times a day. However, the exact dose may be selected from the above range in consideration of age, weight, symptoms and the like.
Hereinafter, a specific example will be described.
[0012]
[Example 1]
(Hydrous ethanol extraction)
1000 g of dry kash was pulverized, 8000 ml of 50% ethanol was added to the powder, and the mixture was stirred occasionally and left at room temperature, and extracted after 24 hours.
The obtained extract was filtered, concentrated under reduced pressure, and further dried by lyophilization to obtain 85.3 g of the target α-amylase inhibitory substance extract.
[0013]
The α-amylase inhibitory activity rate was determined by the following method.
1. α-Amylase [EC: 3.2.2.1.1]
As the α-amylase, α-amylase derived from human saliva (SIGMA, Type IX-A) was used.
[0014]
2. Measurement principle The sample solution is allowed to act on a solution of α-amylase in soluble starch for a certain period of time, and the amount of hydrolyzed starch in the reaction solution is measured.
[0015]
3. Enzymatic reaction (1) Sample solution (50 μl)
Dissolve test extract (extract from Kashiwa) in 50% ethanol.
(2) Substrate buffer (300 μl)
A 0.05M phosphate buffer (pH 7.0) containing 0.04% soluble starch was used.
(3) Enzyme solution (50 μl)
An α-amylase enzyme solution (1 unit, dissolved in 0.05 M phosphate buffer) (pH 7.0) was used.
(4) Color reagent (100 μl)
A 0.01N iodine solution was used.
(5) Diluent (1000 μl)
Purified water was used.
300 μl of substrate buffer was added to 50 μl of the sample solution, and the temperature was equilibrated in a thermostatic bath at 37 ° C. for 5 minutes. Thereafter, 50 μl of enzyme solution was added and reacted at 37 ° C. for 15 minutes. After completion of the reaction, 100 μl of a coloring reagent was added to cause color development, and further 1000 μl of purified water was added for dilution, and the amount of starch produced by hydrolysis was calculated from the absorbance at 660 nm. As a control, 50% ethanol was used instead of the sample solution. In addition, as each blank, what added the enzyme solution after adding the coloring test liquid was used.
[0016]
4). α-Amylase Inhibitory Activity Calculation Method Inhibitory Activity Rate (%) = [((A−B) − (C−D)) / (A−B)] × 100
A: Absorbance of control solution B: Absorbance of control solution blank C: Absorbance of sample solution D: Absorbance of blank of sample solution
The α-amylase inhibitory activity rate (%) in the water-containing ethanol extract obtained in Example 1 was measured, and the results are shown in Table 1. The test sample concentration was shown as the concentration relative to the reaction solution.
[0018]
[Table 1]

[0019]
As shown in Table 1, it was clarified that the Kashou ethanol extract completely inhibited α-amylase activity at a reaction solution concentration of 125 μg / ml, and the degree of inhibition was dependent on the extract concentration. .
[0020]
[Example 2]
(Water extraction)
500 g of dried ash was pulverized, 2500 ml of purified water was added to this powder, the temperature was raised to 95-100 ° C., and extraction was performed for 20 minutes. The obtained extract was filtered, concentrated under reduced pressure, and further dried by a freeze-drying method to obtain 53.6 g of a target α-amylase inhibitor extract.
[0021]
(Calculation of 50% inhibitory activity concentration)
The α-amylase inhibitory activity rate of the katsu extract obtained in Examples 1 and 2 was examined, and the 50% inhibitory activity concentration (IC50: μg · dry weight / ml · reaction solution) was calculated. The IC50 value was calculated and compared using an α-amylase inhibitor derived from wheat seed (SIGMA, Type 1) as an index inhibitor. The respective results are shown in Table 2.
[0022]
[Table 2]

[0023]
As shown in Table 2, in particular, the α-amylase inhibitor containing the water-containing ethanol extract of cashew of the present invention as an active ingredient has an inhibitory activity that is about 6 times higher than that of the wheat seed-derived substance as an indicator inhibitor. It has been shown that it has a sufficiently strong effect even when compared with the natural substance-derived inhibitor material reported so far.
[0024]
[Example 3]
(Formulation)
Using the kashuu extract obtained in Example 1, it was prepared with the following component composition.
10% by weight of Kashuu extract
Lactose 78% by weight
8% by weight of crystalline cellulose
Carboxymethylcellulose calcium 2% by weight
Steering magnesium 2% by weight
The above materials were uniformly mixed, kneaded with purified water, dried, and tablets were produced with a single-type tablet press.
[0025]
[Example 4]
(Granule)
The tablets obtained in Example 3 (tablets) were pulverized, sized and sieved to obtain 10-30 mesh granules.
[0026]
【The invention's effect】
ADVANTAGE OF THE INVENTION According to this invention, the alpha-amylase inhibitor which has the solvent extract from a cashew as a main component is provided. The α-amylase inhibitor obtained by the present invention has higher α-amylase inhibition than conventional α-amylase inhibitors, is effective at low concentrations, and is excellent in safety derived from plant natural products. . Moreover, the α-amylase inhibitor of the present invention can be used for the prevention and improvement of obesity by vigorous dietary therapy by containing it in food.

Claims (1)

An anti-obesity agent comprising, as an active ingredient, a hydrous ethanol (50% ethanol) extract of dried oysters.