Method for extracting antineoplastic active component from Armillaria luteo-virens and application thereof

A technology with anti-tumor activity and Armillaria chrysanthemum, applied in anti-tumor drugs, medical preparations containing active ingredients, plant/algae/fungus/moss ingredients, etc., can solve the lack of quality control standards and lack of effective substances Basic and pharmacodynamic research, difficulty in ensuring the effectiveness of clinical medication, safety and sustainability, etc., to achieve obvious anti-tumor effects, high practicability, and stable properties

Inactive Publication Date: 2016-03-30
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

Since the ingredients of Armillaria armillaria preparations are generally mixtures, there is a lack of necessary material basis and efficacy research, so it lacks reasonable quality control standards, and it is difficult to ensure the effectiveness, safety, and sustainability of clinical medication.

Method used

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  • Method for extracting antineoplastic active component from Armillaria luteo-virens and application thereof
  • Method for extracting antineoplastic active component from Armillaria luteo-virens and application thereof
  • Method for extracting antineoplastic active component from Armillaria luteo-virens and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The method for extracting antitumor active components from Armillaria chrysanthemum in this embodiment comprises the following steps:

[0038] (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.

[0039] (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 ethy...

Embodiment 2

[0048] The method for extracting antitumor active components from Armillaria chrysanthemum in this embodiment comprises the following steps:

[0049] (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.

[0050] (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 f...

Embodiment 3

[0058] Embodiment 3: the experiment of suppressing the growth of human liver tumor Hep-G2 cells

[0059] Cell culture:

[0060] at 5% CO 2 , 37° C., and saturated humidity, human lung tumor Hep-G2 cells were cultured with MEM (10% FBS, 1% PS) medium, and cells in good growth state were selected as experimental cells. After counting the cells, dilute it with medium to obtain a certain concentration of cell suspension.

[0061] Cell growth monitoring:

[0062] Place the cell real-time monitor in 5% CO 2 , 37 ℃ saturated humidity incubator. Take an eight-well plate, add 150 μL of MEM medium to each well, put it into the real-time cell monitor for baseline, take out the eight-well plate after the baseline, and add 345 μL of diluted Hep-G2 cell suspension to each well to reach the number of cells in each well. About 4×10 4 Let stand for 3 minutes, and observe whether the cells are uniform under an inverted microscope. Add 5 μL of diluted medicine (the anti-tumor active compo...

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Abstract

The invention provides a method for extracting an antineoplastic active component from Armillaria luteo-virens. The method comprises the following steps: grinding Armillaria luteo-virens fruiting body into a powder and extracting with ethyl acetate; fetching a supernatant and carrying out one-dimension liquid phase separation: adopting a Silica normal-phase silica gel column and binary organic phases including A phase n-hexane and B phase ethanol, and carrying out gradient elution with concentration of the B phase being 0-75%; collecting 10-14 min eluant and carrying out two-dimension liquid phase separation by the elution mode of isocratic elution with concentration of the B phase being 4-8%; and collecting 6-8 min eluant and carrying out three-dimensional liquid phase separation by the elution mode of isocratic elution with concentration of the B phase being 2-3%, and collecting 7.5-12 min eluant and drying by evaporation so as to obtain a target product. According to the method, separation and purification are carried out by high performance liquid chromatography. Repeatability is high, preparation quantity is large, and operation is simple and time-saving. It is found through experiments that the compound has an inhibitory effect on Hep-g2. Thus, the compound has an anti-cancer drug developing potential and provides theoretical foundation for research and development of new drugs.

Description

technical field [0001] The invention relates to the technical fields of medicinal chemistry and biomedicine, in particular to a method for extracting anti-tumor active components from Armillaria chrysanthemum and the application of the anti-tumor active components in the preparation of anti-liver 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, solid inside. T...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K36/07A61P35/00
CPCA61K36/07A61K2236/33A61K2236/333A61K2236/55
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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