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Compound having alpha-glucosidase inhibitory activity in lotus leaves and application

A technology for glucosidase and inhibitory activity, applied in the field of preparation of drugs for the treatment of diabetes, can solve problems such as difficulty in finding α-glucosidase inhibitors, achieve the goal of reducing blindness, shortening screening time, and improving screening efficiency Effect

Inactive Publication Date: 2012-12-19
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing screening methods for α-glucosidase inhibitors mainly use in vitro enzyme activity inhibition experiments, which can only evaluate the activity of pure compounds, and are difficult to find α-glucosidase inhibitors in complex mixtures such as traditional Chinese medicine

Method used

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  • Compound having alpha-glucosidase inhibitory activity in lotus leaves and application
  • Compound having alpha-glucosidase inhibitory activity in lotus leaves and application
  • Compound having alpha-glucosidase inhibitory activity in lotus leaves and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Preparation of magnetic beads bound to α-glucosidase

[0026] 1. Reagent and solution preparation:

[0027] α-glucosidase (Saccharomyces cerevisiae, simga company); carboxyl magnetic beads Dynabeads® MyOne Carboxylic Acid, specification: 10 mg / ml (invitrogen company); 1-ethyl-3-(3-dimethylaminopropyl) carbon Diimine hydrochloride (EDC, sigma company); 2-(N-morpholine) ethanesulfonic acid (MES, sigma company); N-hydroxysuccinimide (NHS, sigma company).

[0028] Preparation of 25 mmol / L MES solution Weigh an appropriate amount of MES, add water to dissolve, add 1 mol / L sodium hydroxide solution to adjust the pH to 6, and prepare a MES solution with a concentration of 25 mmol / L.

[0029] Preparation of 50 mg / ml EDC solution Weigh an appropriate amount of EDC and quickly dissolve it in cold 25mmol / L MES solution to prepare an EDC solution with a concentration of 50mg / ml. Prepared fresh just before use.

[0030] Preparation of 50 mg / ml NHS solution Weigh an appr...

Embodiment 2

[0040] Example 2 Screening of Compounds with α-Glucosidase Inhibitory Activity in Lotus Leaf Extract

[0041] 1. Preparation of drugs, reagents and solutions:

[0042] Rutin standard (batch number: 100080-200707), National Institute for the Control of Pharmaceutical and Biological Products; Narciscoside standard (batch number: 111209), quercetin standard (batch number: 090426) Shanghai Ronghe Pharmaceutical Technology Development Co., Ltd.; Kaempferol -3-O-β-D-glucuronide standard substance (batch number: P0141), Shanghai Chunyou Biotechnology Co., Ltd.; p-nitrophenyl-α-D-galactopyranoside, sigma company.

[0043] Lotus leaf extract: take lotus leaf and add 70% ethanol to reflux extraction twice, combine the extracts, recover ethanol under reduced pressure, load the concentrated solution on D101 macroporous resin, elute with water, 60% ethanol, and 95% ethanol respectively, discard Remove the eluent from water, combine the 60% ethanol and 95% ethanol eluent, recover the solve...

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Abstract

The invention provides a compound having an alpha-glucosidase inhibitory activity in lotus leaves. Nine compounds having the alpha-glucosidase inhibitory activity are screened from the lotus leaves through magnetic bead separation and liquid chromatogram-mass spectrometry analysis, and the obtained compounds are confirmed to have the alpha-glucosidase inhibitory activity through a verification of the alpha-glucosidase inhibitory activity. Compared with tradition methods, separated compounds are not needed to prepare during a primary screening, thereby substantially improving screening efficiency and shortening screening time. Due to changing a traditional activity-screening mode which separates firstly and screens secondly to a novel activity-screening mode which screens firstly and separates secondly, blindness of preparing and separating compounds is greatly reduced. The method can be popularized and applied to quick screening of compounds having the alpha-glucosidase inhibitory activity from natural medicines or herbal mixtures. The obtained compounds can be used for preparing anti-diabetic drugs.

Description

technical field [0001] The invention belongs to the field of drug screening methods, and relates to a compound with α-glucosidase inhibitory activity in lotus leaves, in particular to the qualitative screening of α-glucosidase from lotus leaves by using magnetic bead separation and liquid chromatography-mass spectrometry. A compound with glucosidase inhibitory activity, and its application in the preparation of medicines for treating diabetes. Background technique [0002] Diabetes is a chronic metabolic disorder, and its main intervention methods are diet control, reduction of ingested carbohydrates, and drug treatment. The carbohydrates ingested by the human body are hydrolyzed by enzymes in the epithelial cells of the small intestine (such as amylase and α-glucosidase) to generate glucose before being absorbed into the blood circulation. Acarbose and other α-glucosidase inhibitors can inhibit α-glucosidase and reduce the conversion of carbohydrates into glucose, which is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H17/07C07H1/08C07D311/30C07D311/40C12Q1/34A61P3/10
Inventor 程翼宇王毅
Owner ZHEJIANG UNIV
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