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A method for separating and purifying paclitaxel from yew branches and leaves or bark

A technology for the separation and purification of yew branches and leaves, which is applied in the field of natural medicine extraction, can solve the problems of complex operation, difficult large-scale production, and low efficiency, and achieve the effects of good repeatability, efficient separation, and easy operation

Inactive Publication Date: 2016-04-20
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because these taxane compounds are very similar to paclitaxel in terms of chemical structure and polarity, the separation of paclitaxel is difficult and the separation and purification process is complicated.
[0003] At present, the separation and purification methods of paclitaxel mainly include: (1) column chromatography, but the operation is relatively complicated, the efficiency is not high, and different fillers are required to be used in combination; (2) preparative high-performance liquid chromatography, which requires high equipment and reagents , the cost is high, and it is difficult to produce on a large scale; (3) high-speed countercurrent chromatography requires a large number of experiments to select a suitable solvent system; (4) supercritical fluid extraction, the equipment cost is high, and the purity of the extracted compound is low

Method used

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  • A method for separating and purifying paclitaxel from yew branches and leaves or bark
  • A method for separating and purifying paclitaxel from yew branches and leaves or bark
  • A method for separating and purifying paclitaxel from yew branches and leaves or bark

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] 1. Treatment of yew branches and leaves

[0064] (1) Take yew branches and leaves and pass through a 24-mesh sieve. Take 500g of powder and add methanol at a ratio of 1:10 (g / mL), soak for 5 hours, ultrasonically extract for 1 hour, and filter to obtain a filtrate. The residue is 1:10 (g / mL). / mL) by adding methanol to repeat the extraction for 1 h, repeat once, and combine the filtrates.

[0065] (2) The filtrate was concentrated under reduced pressure below 60°C to obtain extract (48.75g). The obtained extract was ultrasonically dispersed in water, extracted 3 times with petroleum ether (1:1, v / v); the water layer was obtained after standing and separated, and the water layer was extracted 3 times with ethyl acetate (1:1, v / v ), the ethyl acetate layer was concentrated to dryness under reduced pressure to obtain ethyl acetate extract (3.36g).

[0066] 2. Basic alumina column chromatography

[0067] (1) Take the ethyl acetate extract obtained in step 1, add an appro...

Embodiment 2

[0084] 1. Treatment of yew bark

[0085] (1) Crush the bark of yew bark and pass it through a 24-mesh sieve. Take 500g of powder and add methanol at a ratio of 1:10 (g / mL), soak for 5 hours, reflux for 1 hour, filter to obtain the filtrate, and the residue at a ratio of 1:10 ( g / mL) was added to methanol to repeat the extraction for 1 h, repeated once, and the filtrates were combined.

[0086] (2) The filtrate was concentrated under reduced pressure below 60°C to obtain extract (40.62g). The obtained extract was ultrasonically dispersed in water, extracted 3 times with petroleum ether (1:1, v / v); the water layer was obtained after standing and separated, and the water layer was extracted 3 times with ethyl acetate (1:1, v / v ), the ethyl acetate layer was concentrated to dryness under reduced pressure to obtain ethyl acetate extract (4.26g).

[0087] 2. Basic alumina column chromatography

[0088] (1) Take the ethyl acetate extract obtained in step 1, add an appropriate amou...

Embodiment 3

[0105] 1. Treatment of yew branches and leaves

[0106] (1) Crush the branches and leaves of yew and pass through a 24-mesh sieve, take 500g of powder and add methanol at a ratio of 1:5 (g / mL), soak for 5 hours, reflux for 1 hour, filter to obtain the filtrate, and the residue at a ratio of 1:5 (g / mL) / mL) was added into methanol to repeat the extraction for 1 h, repeated twice, and the filtrates were combined.

[0107] (2) The filtrate was concentrated under reduced pressure below 60°C to obtain extract (45.85g). The obtained extract was ultrasonically dispersed in water, extracted twice with petroleum ether (1:0.5, v / v); the water layer was obtained after standing and separated, and the water layer was extracted twice with ethyl acetate (1:0.5, v / v ), the ethyl acetate layer was concentrated to dryness under reduced pressure to obtain ethyl acetate extract (3.18g).

[0108] 2. Basic alumina column chromatography

[0109] (1) Take the ethyl acetate extract obtained in step...

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Abstract

The invention discloses a method for separating and purifying paclitaxel from yew branches and leaves or bark. The method firstly soaks the branches, leaves or bark of the yew with methanol, ultrasonically extracts or reflux extracts, and concentrates under reduced pressure to obtain the extract; the extract is sequentially extracted with petroleum ether and ethyl acetate to obtain the ethyl acetate extract; Ethyl acetate extract was subjected to basic alumina column chromatography to obtain paclitaxel-rich components, and then treated with methanol-water as a system to obtain crude paclitaxel, and finally used rapid preparative chromatography as a separation method to obtain Methanol-water was used as the elution system, and paclitaxel was finally refined (purity>98.5%). The method of the invention has the advantages of simple operation, low cost, good repeatability, high efficiency and high purity of the obtained product, and can be used for large-scale production of paclitaxel.

Description

technical field [0001] The invention belongs to the technical field of natural medicine extraction. More specifically, it relates to a method for separating and purifying paclitaxel from branches, leaves or bark of Taxus chinensis. Background technique [0002] Southern yew ( Taxuschinensis var. Mairei), also known as beautiful yew, is a plant of the gymnospermae (Gymnospermae), Taxaceae (Taxaceae), and yew genus (Taxus). As one of the most widely distributed yew plants, it is mainly distributed in the Yangtze River Basin, Nanling Mountains and other places. It is the main tree species for the production of anticancer drug Taxol in my country. Paclitaxel is a unique secondary metabolite of Taxus genus. It is one of the most anticancer active natural compounds, and it is also the most popular new anticancer drug in the international market. The mechanism of action of paclitaxel is to promote tubulin polymerization, inhibit depolymerization, maintain tubulin stability, and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D305/14
CPCC07D305/14
Inventor 徐新军梁智坤陶俊妃
Owner SUN YAT SEN UNIV
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