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Medicinal application of timosaponin BIII

A technology of timothy saponin and Anemarrhena chinensis extract is applied in the application field of preparing medicine for treating diabetes, can solve problems such as inability to meet industrial application, and achieve the effects of good anti-diabetic activity and simple preparation method

Inactive Publication Date: 2012-11-07
SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the HPLC separation method is only suitable for laboratories and cannot be satisfied with industrial applications.

Method used

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  • Medicinal application of timosaponin BIII
  • Medicinal application of timosaponin BIII
  • Medicinal application of timosaponin BIII

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1: Extraction and separation method of timosaponin BIII.

[0036] Take 5.0 kg of Anemarrhena medicinal material, degrease by gasoline percolation, and dry. Reflux extraction with 25 liters of 85% ethanol three times for 2 hours each time, filter, and combine the filtrates. Recover the solvent under reduced pressure, concentrate until it has no alcohol smell, and obtain 1 kg of mother extract.

[0037]Suspend the extract with 6 liters of water, and extract four times with 4 liters of n-butanol saturated with water. The n-butanol layer was recovered and dried to obtain 135 grams of extract. The sample was mixed with methanol for silica gel column (2700 g, 160-200 mesh) chromatography. First elute with chloroform until the color is light; then elute with chloroform-n-butanol (1:1). TLC detection, the developing solvent is n-butanol-ethyl acetate-water (4:1:5) upper layer. Using 10% phosphomolybdic acid ethanol at 110oC as the developer, the fractions contai...

Embodiment 2

[0042] Embodiment 2: Extraction and separation method of timosaponin BIII.

[0043] Take 5.0 kg of Anemarrhena medicinal material, degrease by petroleum ether percolation, and dry. Reflux extraction with 40 liters of methanol three times for 2 hours each time, filter, and combine the filtrates. The solvent was recovered under reduced pressure, concentrated until there was no alcohol smell, and 1.2 kg of the mother extract was obtained.

[0044] Suspend the extract with 10 liters of water, and extract four times with 10 liters of n-pentanol saturated with water. The n-pentanol layer was recovered and dried to obtain 150 grams of extract. The sample was mixed with methanol for silica gel column (2700 g, 160-200 mesh) chromatography. First eluted with chloroform until the color became light; then eluted with chloroform-n-butanol (5:1). TLC detection, the developing solvent is n-butanol-ethyl acetate-water (4:1:5) upper layer. Using 10% ethanol phosphomolybdic acid as the dev...

Embodiment 3

[0046] Embodiment 3: In vitro inhibition of α-glucosidase activity test

[0047] a) Reagents and instruments:

[0048] α-glucosidase (EC 232-604-7), purchased from Sigma Company, USA,

[0049] pNPG (4 nitrophenyl-α-D-glucopyranoside) (EC 223-189-3), purchased from Sigma

[0050] Anhydrous sodium carbonate, phosphate, etc., are analytically pure.

[0051] Microplate reader: BIO-TECK Instruments, produced in the United States.

[0052] b) Reagent preparation

[0053] Phosphate buffer (67mM, PH6.8): Weigh an appropriate amount of K 2 HPO 4 ·3H 2 O solution, adjusted to pH 6.8 with phosphoric acid, stored at 4°C until use.

[0054] Enzyme solution: Weigh an appropriate amount of α-glucosidase freeze-dried powder and dilute it with 67mM, pH 6.8 phosphate buffer solution at a concentration of 0.5mg / ml, aliquot 0.5ml into one tube, and freeze at -20°C.

[0055] Substrate PNPG: Prepare 29mM α-PNPG with phosphate buffer (67mM, pH 6.8), aliquot and store at -20°C.

[0056] Reac...

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Abstract

The invention relates to medicinal application of timosaponin BIII, in particular to application of timosaponin BIII serving as an effective ingredient extracted and separated from Chinese medicinal anemarrhena rhizome in preparing a medicament for treating diabetes. Meanwhile, in vivo and in vitro pharmacological experiments prove that the compound has remarkable inhibiting activity on alpha-glucosidase, can effectively reduce blood sugar of animals, and can be applied in preparing an alpha-glucosidase inhibitor and preparing the medicament for treating the diabetes. The timosaponin BIII compound shows good diabetes activity resistance in vivo, develops the application field thereof, and may become an effective medicament for treating the diabetes in the future.

Description

[0001] The patent application of the present invention is a further divisional application following the application date of May 16, 2007, the application number is 2007100407304, and the invention title is "Preparation Method and Application of Timosaponin BIII". technical field [0002] The invention relates to the use of plant extracts, in particular to the application of timosaponin BIII, an active ingredient extracted and separated from the traditional Chinese medicine Zhimu, in the preparation of medicines for treating diabetes. Background technique [0003] Diabetes is a common clinical disease caused by the absolute or relative lack of insulin in the human body. The concentration of glucose in the blood increases, and then a large amount of sugar is excreted from the urine, resulting in polydipsia, polyuria, polyphagia, weight loss, dizziness, and fatigue. and other symptoms. Further development then causes various serious acute and chronic complications throughout t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/7048A61P3/10
Inventor 黄成钢马春辉唐意红李志雄赵维民范明松朱海燕孙兆林施松
Owner SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI
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