Screening method and application of alpha-glucosidase inhibitor

A technology of glucosidase and screening method, which is applied in the application field of traditional Chinese medicine extracts, can solve the problems of poor reproducibility, large consumption of reagents, and no activity, etc., and achieve the effect of simple, reasonable and easy-to-operate screening method

Inactive Publication Date: 2011-02-16
DALIAN POLYTECHNIC UNIVERSITY
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Problems solved by technology

At present, the in vitro screening methods for α-glucosidase inhibitors mainly include p-nitrophenyl-α-D-glucopyranoside (PNPG) method, glucose oxidase method and immobilized enzyme method, but most of them use spectrophotometry As a detecti

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  • Screening method and application of alpha-glucosidase inhibitor
  • Screening method and application of alpha-glucosidase inhibitor
  • Screening method and application of alpha-glucosidase inhibitor

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[0021] The present invention is described in detail through the following specific implementations, but is not limited to the following examples.

[0022] 1. Establishment of an in vitro screening model for α-glucosidase inhibitors

[0023] (1) Determination of α-glucosidase inhibitory activity: the experiment was carried out in a 1.5 mL centrifuge tube, and the total reaction volume was 160 μL: 10 μL 0.067 mol·L -1 , pH 6.8 phosphate buffer solution, 30 μL concentration 0.2 U mL -1 α-Glucosidase, shake and mix, incubate at 37 °C for 20 min, add 40 μL of 8.0 mM PNPG to start the reaction, shake and mix, after 30 min of reaction at 37 °C, add 80 μL of 0.2 M Na 2 CO 3 To terminate the reaction, add ultrapure water to dilute to 500 μL, shake and mix well, filter through a 0.45 μm membrane, and detect by HPLC, and calculate the activity of α-glucosidase by the concentration change of the product PNP. The principle of α-glucosidase hydrolysis of substrate PNPG:

[0024]

[...

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Abstract

The invention belongs to the field of traditional Chinese medicines. The screening method of an alpha-glucosidase inhibitor comprises the following steps of: (1) establishing in vitro screening models; establishing in vitro screening models of alpha-glucosidase and a substrate PNPG (Para Nitrobenzene-Beta-D-Galactose Pyranoside); (2) establishing a component library: selecting traditional Chinesemedicines, systematically extracting each traditional Chinese medicine with various (more than two kinds) solvents according to a similar compatible principle according to the polarities from low to high, wherein multiple extracting parts can be obtained for each traditional Chinese medicine by adopting the method; and (3) screening: blending all the extracting parts of the traditional Chinese medicines into a solution with a certain concentration, adding the solution to a reaction model, and calculating the inhibition rate. The screening method is simple, reasonable and easy to operate and can be used for finally screening out traditional Chinese medicines and traditional Chinese medicine components having obvious inhibition effects on the alpha-glucosidase. In addition, the screened traditional Chinese medicines and the traditional Chinese medicine components can be used for developing products aiming to inhibit the alpha-glucosidase and particularly used for developing drugs for reducing the blood sugar and has industrialization significance.

Description

technical field [0001] The invention belongs to the field of traditional Chinese medicines, relates to a screening method for active components of traditional Chinese medicines inhibiting α-glucosidase, and also relates to the application of traditional Chinese medicine extracts. Background technique [0002] With the improvement of people's living standards and the change of dietary structure, the number of diabetic patients increases rapidly and shows a trend of younger age. Diabetes mellitus (DM) has become one of the major chronic diseases that seriously threaten human health. The two important indicators of diabetes detection are fasting blood sugar level and postprandial blood sugar level. Epidemiological studies have shown that the first symptom in the onset of diabetes is often postprandial hyperglycemia, and postprandial hyperglycemia is more likely to cause than fasting blood sugar. Various complications are also an important factor in the development of type Ⅱ dia...

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Application Information

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IPC IPC(8): A61K36/486A61K36/71A61K36/13A61K36/076A61K36/8969A61K36/70A61K36/8945G01N30/02A61P3/10A61K36/739A61K36/63A61K36/605A61K36/61A61K36/536A61K36/37A61K36/484A61K36/8964A61K36/54A61K36/21A61K36/815A61K35/64A61K36/708A61K36/28A61K36/185A61K36/8984A61K36/39
Inventor 朱靖博朱文佳付绍平许芹永
Owner DALIAN POLYTECHNIC UNIVERSITY
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