Preparation method for 3-surface ester bufotalin

Inactive Publication Date: 2012-07-04
CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, at present, the preparation of 3-epibufagenin usually uses chemical methods (GEORGER. PETTIT, Y.A.. J. Org. Chem 37 (25): 4040-4044.), the process is cumbersome, produces a large number of by-products, and the preparation process Most of them use benzene and other organic solvents that are highly toxic to humans, which limits the production and application of 3-epibufagenin

Method used

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  • Preparation method for 3-surface ester bufotalin
  • Preparation method for 3-surface ester bufotalin
  • Preparation method for 3-surface ester bufotalin

Examples

Experimental program
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Effect test

Embodiment 1

[0039] Embodiment 1: GAM nutrient solution preparation

[0040] Tryptone 10g, Peptone 10g, Soytone 3g, Beef Extract 2.2gL, Glucose 3g, Potassium Dihydrogen Phosphate 2.5g, Sodium Chloride 5g, Starch 5g, L-cysteine ​​0.5g, Sodium Thioglycolate 0.3 g. Dissolve 5 g of yeast extract and 1.2 g of bovine liver extract powder in 800 mL of distilled water, adjust the pH value to 7.1-7.2, adjust the volume to 1 L, and sterilize at 120°C for 20 minutes.

Embodiment 2

[0041] Embodiment 2: normal rat feeding

[0042]Select 10 Wistar rats with a body weight of 220-250g, half male and half male, under the conditions of temperature 18-26°C, humidity 40-70%, and light 12-14h / day, commercialized in granular or pellet form Stir-fed rodent diet and tap water.

Embodiment 3

[0043] Example 3: Preparation and purification of 3-epibufagenin

[0044] Take 0.2 g of fresh rat feces and add it to 10 mL of GAM culture solution, and culture it anaerobically at 37° C. for 24 h. Then 1.6 mg of bufagenin was added, and anaerobic culture was carried out at 37° C. for 43 hours to obtain a culture solution. The culture solution was extracted with 100 times the volume of ethyl acetate, the extract was filtered through a microporous membrane, and the 3-epiester bufogenin standard was used as a control, and analyzed by high performance liquid chromatography and mass spectrometry.

[0045] The detection conditions of high performance liquid chromatography are Waters 2695 chromatograph; Zorbax Eclipse XDB-C18 chromatographic column (4.6mm×150mm, Agilent); mobile phase A is acetonitrile, B is an aqueous solution containing 0.3% acetic acid; flow rate: 0.3mL / min; Column temperature: 35°C; Gradient: 0-25min A is 20%-80%, 25-35min A keeps 80% unchanged. The result is ...

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Abstract

The invention relates to the field of microorganism transformation and discloses a preparation method for 3-surface ester bufotalin, which comprises the steps of: anaerobically culturing suspension containing rat intestinal flora in GAM culture liquid at the temperature of 37 DEG C for 22 to 25 hours; adding ester bufotalin in the culture liquid to anaerobically culture for 40 to 80 hours; taking the culture liquid to be extracted with a non-polar solvent; and collecting the extraction liquid to obtain 3-surface ester bufotalin. 0.1 to 0.3mg ester bufotalin is into every l-milliliter culture liquid. The preparation method utilizes rat intestinal flora to transform the ester bufotalin to prepare the 3-surface ester bufotalin, thereby being simple in method, small in byproducts generated during preparation processes, high in ester bufotalin transformation rate, high in purity of the prepared 3-surface ester bufotalin and capable of being applied to large preparation of the 3-surface ester bufotalin.

Description

technical field [0001] The invention relates to the field of microbial transformation, in particular to a preparation method of 3-epibufagenin. Background technique [0002] Toad venom is a traditional Chinese medicinal material, which is processed from the secretions of the post-auricular glands and skin glands of amphibians and reptiles of the Bufo family. Toad Su is sweet, pungent, warm in nature, poisonous, contains steroids, alkaloids and other biologically active substances, and has multiple functions such as detoxification, analgesia, resuscitation, and antitumor. [0003] Toad venom is a traditional medicinal material in my country, and its sources of medicine are relatively rich. Resibufogenin (resibufogenin) is the main active ingredient in the traditional Chinese medicine Chansu. It is a steroidal compound with an α-pyrone ring at the C-17 position. Its molecular formula is C 2 4H 32 o 4 , the molecular weight is 384.51, and the structural formula is as follow...

Claims

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Application Information

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IPC IPC(8): C12P17/16
Inventor 刘志强韩天娇宋凤瑞郑重邢俊鹏刘淑莹
Owner CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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