Method for producing 7-xylose-10-deacetyl paclitaxel and special culture medium thereof

A technology for deacetylpaclitaxel and culture medium, which is applied in the field of producing 7-xylose-10-deacetylpaclitaxel, and achieves the effects of short growth cycle, environmental resource protection and low cost

Active Publication Date: 2014-07-23
ZHANGJIAGANG IND TECH RES INST CO LTD DALIAN INST OF CHEM PHYSICS CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, it should be an effective method to obtain 7-xylose-10-deacetylpaclitaxel by plant cell culture, but unfortunately there is no report in this regard before

Method used

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  • Method for producing 7-xylose-10-deacetyl paclitaxel and special culture medium thereof
  • Method for producing 7-xylose-10-deacetyl paclitaxel and special culture medium thereof
  • Method for producing 7-xylose-10-deacetyl paclitaxel and special culture medium thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Obtaining 7-xylose-10-deacetylpaclitaxel by culturing the tender stems of Taxus chinensis

[0024] Basic medium: NH 4 NO 3 1300mg, KNO 3 2000mg, KH 2 PO 4 360mg, CaCl 2 2H 2 O 460mg, MgSO 4 ·7H 2 O 380mg, FeSO 4 ·7H 2 O 29 mg, Na 2 -EDTA 38mg, MnSO 4 4H 2 O 2.5 mg, H 3 BO 3 0.7mg, KI 0.9mg, Na 2 MoO 4 2H 2 O 0.03mg, CuSO 4 ·5H 2 O 0.003mg, CoCl 2 ·6H 2 O 0.003mg, Zn.Na-EDTA 17mg, thiamine hydrochloride 0.6mg, nicotinic acid 0.6mg, pyridoxine hydrochloride 0.6mg, glycine 3mg, dilute to 1L with water, pH5.8.

[0025] Callus induction medium: 7 mg of 2,4-D, 50 g of sucrose, and 8 g / L of agar are added to the basic medium.

[0026] Subculture medium: 5 mg of 2,4-D, 50 g of sucrose, and 8 g / L of agar were added to the basic medium.

[0027] The sterilization conditions of the above-mentioned culture medium are all: sterilization at 121° C. for 20 minutes.

[0028] The method of the present invention is used to cultivate Taxus chinensis, ...

Embodiment 2

[0037] Example 2: Obtaining 7-xylose-10-deacetylpaclitaxel by culturing the needles of Taxus yunnanensis

[0038] Basic medium: NH 4NO 3 1200mg, KNO 3 1900mg, KH 2 PO 4 340mg, CaCl 2 2H 2 O 440mg, MgSO 4 ·7H 2 O 370mg, FeSO 4 ·7H 2 O 27.8 mg, Na 2 -EDTA 37.3mg, MnSO 4 4H 2 O 2.23 mg, H 3 BO 3 0.63mg, KI 0.83mg, Na 2 MoO 4 2H 2 O 0.025mg, CuSO 4 ·5H 2 O 0.0025 mg, CoCl 2 ·6H 2 O 0.0025mg, Zn.Na-EDTA 15mg, thiamine hydrochloride 0.5mg, nicotinic acid 0.5mg, pyridoxine hydrochloride 0.5mg, glycine 2mg, dilute to 1L with water, pH5.8.

[0039] Callus induction medium: add 2,4-D 6mg, sucrose 40g, and agar 6g / L on the basis of basic medium.

[0040] Subculture medium: 4 mg of 2,4-D, 40 g of sucrose, and 6 g / L of agar were added to the basic medium.

[0041] The sterilization conditions of the above-mentioned culture medium are all: sterilization at 121° C. for 20 minutes.

[0042] Now use the method of the present invention to cultivate Taxus yunnanensis,...

Embodiment 3

[0051] Example 3: Obtaining 7-xylose-10-deacetylpaclitaxel by culturing the male flowers of Taxus sinensis

[0052] Basic medium: NH 4 NO 3 1100mg, KNO 3 1800mg, KH 2 PO 4 320mg, CaCl 2 2H 2 O 420mg, MgSO 4 ·7H 2 O 350mg, FeSO 4 ·7H 2 O 26 mg, Na 2 - EDTA 36mg, MnSO 4 4H 2 O 2.0 mg, H 3 BO 3 0.5mg, KI 0.7mg, Na 2 MoO 4 2H 2 O 0.02mg, CuSO 4 ·5H 2 O 0.002mg, CoCl 2 ·6H 2 O 0.002mg, Zn.Na-EDTA 12mg, thiamine hydrochloride 0.4mg, nicotinic acid 0.4mg, pyridoxine hydrochloride 0.4mg, glycine 1mg, dilute to 1L with water, pH5.6.

[0053] Callus induction medium: add 2,4-D 6mg, sucrose 20g, and agar 5g / L on the basis of basic medium.

[0054] Subculture medium: 1 mg of 2,4-D, 20 g of sucrose, and 5 g / L of agar were added to the basic medium.

[0055] The sterilization conditions of the above-mentioned culture medium are all: sterilization at 121° C. for 20 minutes.

[0056] Now use the method of the present invention to cultivate Taxus sinensis, and extra...

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Abstract

The invention provides a method for producing 7-xylose-10-deacetyl paclitaxel and a special culture medium thereof. By adopting the culture method and the special medium, a callus for producing the 7-xylose-10-deacetyl paclitaxel can be obtained, and the content of the 7-xylose-10-deacetyl in the callus is 0.005-0.009 percent. For the method, the 7-xylose-10-deacetyl in a yew plant is firstly obtained through a cell engineering method. The produced 7-xylose-10-deacetyl paclitaxel can be used as a raw material to be converted into paclitaxel through a simple process. The method solves the problem of raw materials on the production of paclitaxel, is easy to operate and low in cost, has a short cycle, and can protect yew plant resources.

Description

technical field [0001] The invention belongs to the field of plant cell engineering, and specifically relates to a method for producing 7-xylose-10-deacetylpaclitaxel and a special culture medium thereof. Background technique [0002] Paclitaxel (1), as a first-line anticancer drug, is currently widely used in the treatment of various solid cancers such as ovarian cancer, breast cancer, and lung cancer. At present, the way of large-scale production of paclitaxel is mainly to directly extract or extract its semi-synthetic precursor, 10-deacetylbaccatin III (2), from Taxus plants. [0003] [0004] Paclitaxel was synthesized by synthetic means. Recently, using 7-xylose-10-deacetylpaclitaxel (3) as raw material, the process of synthesizing paclitaxel has been successively developed (US5412116, US6028206, US5856532, US6437154, ZL200610046296.6 and ZL200610047743.X). 7-xylose-10-deacetylpaclitaxel and paclitaxel belong to tricyclic diterpenoids, which are widely found in the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/04
Inventor 杨凌侯琨栾宏伟
Owner ZHANGJIAGANG IND TECH RES INST CO LTD DALIAN INST OF CHEM PHYSICS CHINESE ACADEMY OF SCI
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