Method for extracting cordycepin component from Cordyceps militaris and application of cordycepin component

A technology of cordycepin and Cordyceps militaris, which is applied in the field of application of cordycepin components in the preparation of anti-tumor drugs, can solve the problems of complex extraction and purification methods of cordycepin, achieve objective experimental results, simple experimental operations, and small human errors Effect

Active Publication Date: 2014-05-21
贵州贵安精准医学股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In recent years, the research of Cordyceps militaris has become a hot spot, especially the research of cordycepin in Cordyceps militaris, but the extraction and purification methods of cordycepin are relatively complicated

Method used

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  • Method for extracting cordycepin component from Cordyceps militaris and application of cordycepin component
  • Method for extracting cordycepin component from Cordyceps militaris and application of cordycepin component
  • Method for extracting cordycepin component from Cordyceps militaris and application of cordycepin component

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Extraction of Cordycepin militaris components

[0034] Extracting the cordycepin component from Cordyceps militaris specifically includes the following steps:

[0035] (1) After the fruiting body of Cordyceps militaris was dried in the shade at room temperature, it was dried and dehydrated at 70°C, and then pulverized in an ultrafine pulverizer at -20°C for 1 min to obtain ultrafine powder;

[0036] (2) Take 3Kg of the superfine powder of the above-mentioned Cordyceps militaris fruiting bodies, and boil them three times: the first boil: add 3kg of Cordyceps militaris powder to 24L ultrapure water, boil for 70min, after boiling, centrifuge at 25°C and 6000rpm for 30min, respectively Collect the supernatant and residue, transfer the supernatant to a constant temperature blast drying oven at 60°C to dry and solid; the second boiling: add 12L ultrapure water to the residue from the first boiling, boil for 70 minutes, press the The first water boiling method is u...

Embodiment 2

[0040] Embodiment 2: the test of inhibiting the growth of human lung cancer A549 cells

[0041] Cell culture:

[0042] at 5%C0 2 , 37°C, under saturated humidity, culture human lung cancer A549 cells in F-12K (10% FBS + 1% PS) medium, and select the cells growing in the logarithmic phase as experimental cells. After counting the cells, dilute it with medium to obtain a certain concentration of cell suspension.

[0043] Cell growth monitoring:

[0044] Place the cell real-time monitor in 5% CO 2 , 37°C saturated humidity incubator. Take an 8-well plate, add 150 μL of F-12K medium to each well, and put it into the real-time cell monitor for baseline. After the baseline, take out the eight-well plate and add 345 μL of diluted A549 cell suspension to each well to reach the number of cells in each well. About 2×10 4 Let stand for 3 minutes, and observe whether the cells are uniform under an inverted microscope. Add 5 μL of the diluted drug (the cordycepin component extracted...

Embodiment 3

[0049] Embodiment 3: the test of suppressing the growth of human liver cancer Hep-G2 cells

[0050] Cell culture:

[0051] at 5%C0 2 , 37°C, under saturated humidity, cultured human lung cancer Hep-G2 cells in DMEM (10% FBS, 1% PS) medium, and selected cells with good growth status as experimental cells. After counting the cells, dilute it with medium to obtain a certain concentration of cell suspension.

[0052] Cell growth monitoring:

[0053] Place the cell real-time monitor in 5% CO 2 , 37°C saturated humidity incubator. Take an 8-well plate, add 150 μL of MEM medium to each well, put it into a 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 the diluted drug (the cordycepin component extracted in Example ...

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Abstract

The invention provides a method for extracting a cordycepin component from Cordyceps militaris. The method comprises the following steps: ultrafinely pulverizing Cordyceps militaris fruiting body to obtain ultrafine powder; extracting the ultrafine powder with ultrapure water, collecting supernatant, drying, precipitating with ethanol, centrifuging, collecting supernatant, performing rotary evaporation, and drying to obtain crude extract; dissolving the crude extract with 15% methanol-water solution to 100 mg/mL, separating with preparative liquid phase DAC reversed phase chromatographic column with reversed phase separation filler of C18, performing gradient elution with 5-15% methanol-water solution, performing isocratic elution with 15% methanol-water solution, setting detection wavelength of 260 nm, performing in vitro antitumor activity screening of components, and selecting the component with the highest activity, i.e. target cordycepin component. The method provided by the invention is simple and controllable, and has good repeatability; the content of cordycepin in the obtained component is high, the batch quality is stability, and the cordycepin component can be used for preparing antitumor drugs.

Description

technical field [0001] The invention relates to the technical field of medical biological preparation, in particular to a method for extracting cordycepin components from Cordyceps militaris, and the application of the extracted cordycepin components in the preparation of antitumor drugs. Background technique [0002] Cordycepin is a secondary metabolite produced by Cordyceps militaris, and in vitro experiments have been proven to be nucleoside analogs with a wide range of biological activities and pharmacological effects. In 1951, Cunningham et al. in Canada isolated a substance from the original slurry of Cordyceps militaris, named Cordycepin (Cordycepin), which is a nucleotide composed of adenosine and deoxypentose with a carbon branch, so it is also Known as 3'-deoxyadenosine, it is the first deoxynucleoside antibiotic isolated from fungal organisms, a nucleic acid derivative of nitrogen-containing glycosides, and belongs to the purine alkaloids. [0003] As early as th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K36/068A61P35/00
Inventor 张耀洲张晓倩崔今松
Owner 贵州贵安精准医学股份有限公司
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