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Preparation method of high-purity paclitaxel

A paclitaxel and high-purity technology, which is applied in the field of cell culture of medicinal plants, can solve the problems of low content, low extraction purity, and difficult extraction, and achieve the effects of stabilizing cell lines, prolonging the time of browning, and increasing growth speed

Inactive Publication Date: 2019-01-18
覃家日
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the prior art, the source of paclitaxel is obtained from the bark of Taxus chinensis, the content is very low, the extraction is difficult, and the extraction purity is low, which cannot meet people's growing demand for high-purity paclitaxel at all.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] A preparation method of high-purity paclitaxel, which comprises the following steps:

[0025] 1) Select the buds on the tender stems of Taxus Mandia as culture materials, wash them with detergent and tap water, rinse them under running water for 10 minutes, place them in 75% alcohol for 10 seconds, and then wash them with 0.1 % mercuric chloride, washed with sterile water after disinfection, cut off the discolored part of the explant, and then carried out induction culture, and selected fresh embryogenic callus with vigorous growth, loose texture, stable and uniform state for three consecutive subcultures to inoculate Into the liquid suspension medium, shaking culture, subculture once every 7 days, after 2 months of subculture, establish a stable growth yew cambium stem cell culture system;

[0026] 2) Add the yew endophytic fungus extract to the suspension culture medium, mix evenly to obtain a mixed culture solution, place the mixed culture solution in a bioreactor, a...

Embodiment 2

[0032] A preparation method of high-purity paclitaxel, which comprises the following steps:

[0033]1) Select the buds on the tender stems of Taxus chinensis as culture materials, wash them with detergent and tap water, rinse them under running water for 10 minutes, place them in 75% alcohol for 10 seconds, and then pass them through 0.1% liters of water. After mercury disinfection, rinse with sterile water, cut off the discolored part of the explant, and then conduct induction culture, select fresh embryogenic callus that has been subcultured for 3 times and has vigorous growth, loose texture, stable and uniform state, and inoculated into the liquid suspension. In the culture medium, shake culture, subculture once every 7 days, and establish a stably growing taxus cambium stem cell culture system after 2 months of subculture; wherein, a germination bed is set in the culture dish, and the germination bed is from the top The first layer to the bottom is filter paper and the sec...

Embodiment 3

[0040] A preparation method of high-purity paclitaxel, which comprises the following steps:

[0041] 1) Select the buds on the tender stems of Taxus chinensis as culture materials, wash them with detergent and tap water, rinse them under running water for 10 minutes, place them in 75% alcohol for 10 seconds, and then pass them through 0.1% liters of water. After mercury disinfection, rinse with sterile water, cut off the discolored part of the explant, and then conduct induction culture, select fresh embryogenic callus that has been subcultured for 3 times and has vigorous growth, loose texture, stable and uniform state, and inoculated into the liquid suspension. In the culture medium, shake culture, subculture once every 7 days, and establish a stably growing taxus cambium stem cell culture system after 2 months of subculture; wherein, a germination bed is set in the culture dish, and the germination bed is from the top The first layer to the bottom is filter paper and the se...

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Abstract

The invention provides a preparation method of high-purity paclitaxel. The preparation method comprises the following steps: (1) establishing a stable taxus chinensis forming layer stem cell culture system; (2) adding taxus chinensis endophytic fungi extract liquid into a suspension culture medium, inoculating taxus chinensis cells, performing the ventilation culture for 6 to 8 days under an artificial lighting condition, and adding jasmonic acid methyl ester and a clinacanthus nutans extract into the liquid suspension culture medium to continue the culture; (3) collecting the taxus chinensiscells, and performing the coarse extraction of paclitaxel; (4) purifying a crude extract; and (5) enabling a concentrated solution II to pass through a C18 high-performance liquid phase chromatographic purification column, eluting by using methanol, collecting and combining paclitaxel elution parts, decompression evaporating the solvent methanol, crystallizing and releasing the paclitaxel, centrifugally collecting crystals, collecting and combining the paclitaxel elution part, decompression evaporating the solvent methanol to crystallize and precipitate the paclitaxel, centrifugally collectingthe crystals, and vacuum freeze drying the crystals, thus obtaining the high-purity paclitaxel concentrated solution II. The preparation method has the characteristics of high induction success rate,high paclitaxel extraction purity and the like.

Description

technical field [0001] The invention relates to the technical field of cell culture of medicinal plants. More specifically, the present invention relates to a preparation method of high-purity paclitaxel. Background technique [0002] Taxus is a class of trees and shrubs belonging to the genus Taxus in the family Taxaceae, and there are 11 species in the world. Yew has a wide distribution range, from the subarctic regions of Eurasia and North America to the subtropical and even tropical regions of Central America and Southeast Asia. Studies have shown that paclitaxel in yew has special effects on the treatment of ovarian cancer, lung cancer, colon cancer, metastatic breast cancer, melanoma, leukemia and other cancers, making paclitaxel one of the most popular anticancer drugs at present. However, in the prior art, the source of paclitaxel is obtained from the bark of Taxus chinensis, the content is very low, the extraction is difficult, and the extraction purity is low, wh...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D305/14
CPCC07D305/14
Inventor 覃家日
Owner 覃家日
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