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Method of extracting ergosterol peroxide from armillaria luteo-virens and application of ergosterol peroxide

A technology of ergosterol peroxide and Armillaria yellow-green, applied in the directions of steroids, organic chemistry, drug combination, etc., can solve the problems of cumbersome extraction steps, low repeatability, small preparation amount and the like, and achieves the preparation amount Large, repeatable, short time period effects

Inactive Publication Date: 2016-05-18
正源堂(天津)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the extraction steps of this compound are cumbersome, the reproducibility is not high, and the yield is low, and the preparation amount is small, which can only meet the needs of laboratory research.
However, the research on this compound is mostly used in biochemistry, and there are few reports on its anticancer pharmacological activity.

Method used

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  • Method of extracting ergosterol peroxide from armillaria luteo-virens and application of ergosterol peroxide
  • Method of extracting ergosterol peroxide from armillaria luteo-virens and application of ergosterol peroxide
  • Method of extracting ergosterol peroxide from armillaria luteo-virens and application of ergosterol peroxide

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] The present embodiment extracts the method for ergosterol peroxide from Armillaria chrysanthemum, comprises the following steps:

[0046] (1) Armillaria chrysanthemum fruiting body is used as raw material, dried in the shade at room temperature, dried and dehydrated at 70°C, and then ultrafinely pulverized at -20°C for 1 min to obtain Armillaria chrysanthemum ultrafine powder.

[0047] (2) Take 3kg of the above-mentioned Armillaria chrysanthemum superfine powder, soak it in ethyl acetate three times, soak for the first time: add 15L ethyl acetate to 3kg Armillaria chrysanthemum superfine powder, soak for 48h at 25°C, after soaking, 20 Centrifuge at 4000rpm for 30min, and collect the supernatant and residue respectively. The second soaking in ethyl acetate: Add 15L of ethyl acetate to the first residue, soak at 25°C for 36h, after soaking, centrifuge at 20°C and 4000rpm for 30min, and collect the supernatant and residue respectively. The third soaking: add 15L ethyl ace...

Embodiment 2

[0055] The present embodiment extracts the method for ergosterol peroxide from Armillaria chrysanthemum, comprises the following steps:

[0056] (1) Armillaria chrysanthemum fruiting body is used as raw material, dried in the shade at room temperature, dried and dehydrated at 70°C, and then ultrafinely pulverized at -20°C for 1 min to obtain Armillaria chrysanthemum ultrafine powder.

[0057] (2) Take 5kg of the above-mentioned Armillaria chrysanthemum superfine powder, soak it through ethyl acetate three times, soak for the first time: add 25L ethyl acetate to 5kg Armillaria chrysanthemum superfine powder, soak for 48h at 25°C, after soaking, 20 Centrifuge at 4000rpm for 30min at ℃, collect the supernatant and residue respectively, concentrate the supernatant by rotary evaporation and transfer to a constant temperature blast drying oven at 60℃ to dry to constant weight. The second soaking in ethyl acetate: add 25L of ethyl acetate to the first residue, and soak at 25°C for 36...

Embodiment 3

[0064] Example 3: Confirmation of ergosterol peroxide

[0065] 1. Purity analysis by HPLC:

[0066] The object component that embodiment 1 obtains ( image 3 5-1-2 peak) was analyzed by HPLC. It is a single compound with a purity of more than 98% detected by high-performance liquid phase, and the HPLC spectrum is shown in Figure 7 .

[0067] 2. Mass spectrometry analysis:

[0068] For the purpose component ( image 3 Middle 5-1-2 peak) for mass spectrometry analysis: Hanmeng Biotechnology (Tianjin) Co., Ltd., mass spectrogram see Figure 8 . EI-MSm / z: 467(M+K), 429(M+H) + .

[0069] 3. NMR analysis:

[0070] For the purpose component ( image 3 Middle 5-1-2 peak) carry out nuclear magnetic resonance analysis: by carbon spectrum, hydrogen spectrum, COZY correlation spectrum (such as Figure 9-12 ) evidence analysis obtained the following results:

[0071] 1H-NMR (CDCl3, 400MHz): δ3.95(1H, m, H-3), 6.26(1H, d, J=8.5Hz, H-6), 6.52(1H, d, J=8.5Hz, H -7),0.83(3H,s,H-18)...

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Abstract

The invention provides a method of extracting ergosterol peroxide from armillaria luteo-virens. The method comprises the steps that armillaria luteo-virens sporocarps are used as a raw material, ground into powder and then subjected to ethyl acetate extraction; supernatant is taken for one-dimensional liquid phase separation, a Silica normal-phase silica gel chromatographic column and binary organic phases of A-phase normal hexane and B-phase ethyl alcohol are adopted, and the B-phase concentration is subjected to gradient elution from 0% to 75%; a one-dimensional component is obtained for two-dimensional liquid phase separation, and the elution mode is changed to the mode that the B-phase concentration is subjected to isocratic elution to 4%-8%; a two-dimensional component is obtained for three-dimensional liquid phase separation, the elution mode is changed to the mode that the B-phase concentration is subjected to isocratic elution to 2%-3%, and the target product is obtained. According to the method, efficient liquid phase chromatography is adopted for separation and purification, the separation method is stable, repeatability is high, the prepared amount is large, operation is simple, and time is saved. Experiments find that the compound has an inhibitory effect on Hep-g2 and A549, thereby having the potential of being developed into an anticancer drug and providing a theoretical basis for new drug research and development.

Description

technical field [0001] The invention relates to the technical fields of medicinal chemistry and biomedicine, in particular to a method for extracting ergosterol peroxide from Armillaria chrysanthemum, and the application of the ergosterol peroxide in the preparation of anti-liver cancer drugs and anti-lung cancer drugs . Background technique [0002] Armillaria luteo-rivens, also known as chanterelles, golden mushrooms, and yellow rings, belongs to Basidiomycotina, Phytomycetes, Agaricaceae, White Mushrooms, and Armillaria. The fruiting body of Armillaria chrysogenum is medium and large, the cap is flat hemispherical to flat, with a diameter of 5-12cm, the edge of the cap is involuted, and the epidermis is cracked to form ciliated posterior phosphorus sheets arranged in a near ring. The gills are similar to the cap color, slightly dense, curved, and unequal in length. The stipe is cylindrical, the stem is enlarged, 2-10cm long, 2-2.5cm in diameter, white or yellowish, soli...

Claims

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Application Information

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IPC IPC(8): C07J71/00A61P35/00
CPCC07J71/0005
Inventor 张耀洲王欢欢杜思邈盖其静
Owner 正源堂(天津)生物科技有限公司
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