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Membrane extraction separation technique of epothilone

A technology of epothilone and membrane extraction, applied in the direction of organic chemistry, etc., can solve the problems of inefficient separation of epothilone a, b, small extraction amount, etc., to reduce product cost, operation difficulty, and use amount , the effect of improving product quality

Inactive Publication Date: 2014-09-10
虞龙
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Biological fermentation mainly produces epothilone a and epothilone b, and its filtration is generally through resin analysis, but the expensive equipment, the extraction amount is small, and epothilone a and b cannot be separated efficiently, which has become a One of the factors restricting the industrial production of epothilones a and b

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Filter 100ml of the fermentation broth with epothilone a and b content of 550mg / L detected by HPLC through an organic membrane, and the specification of the organic membrane is 5000 molecular weight cut-off, to obtain a total of 103ml of filtrate and lotion, and then pass the filtrate and lotion through ceramic Membrane filtration, the specification of the ceramic membrane is 1000 molecular weight cut-off, and the filtrate and lotion are 76ml in total, and the lotion is ordinary distilled water, and the collected filtrate and lotion are eluted through a SephadexG-15 gel chromatographic column, and the eluent is: acetone It is prepared with petroleum ether at a volume ratio of 1:7. HPLC detects the eluent, collects the fractions whose total chromatographic purity of epothilone a and b reaches more than 80%, obtains 58ml of eluent, and then elutes Concentrate the solution through a nanofiltration membrane with a molecular weight cut-off of 500 to obtain 48ml of a concentra...

Embodiment 2

[0027] Filter 100ml of the fermentation broth with epothilone a and b content of 550mg / L detected by HPLC through an organic membrane, and the specification of the organic membrane is 5500 molecular weight cut-off, to obtain a total of 98ml of filtrate and lotion, and then pass the filtrate and lotion through ceramic Membrane filtration, the specification of the ceramic membrane is 1000 molecular weight cut-off, and the filtrate and lotion are obtained in total 75ml, and the lotion is ordinary distilled water, and the collected filtrate and lotion are eluted through a SephadexG-15 gel chromatography column, and the eluent is: acetone It is prepared with petroleum ether at a volume ratio of 1:7. HPLC detects the eluent, collects the fractions whose total chromatographic purity of epothilone a and b reaches more than 80%, obtains 52ml of eluent, and then elutes Concentrate the solution through a nanofiltration membrane with a molecular weight cut-off of 500 to obtain 43ml of a co...

Embodiment 3

[0030] Filter 100ml of the fermentation broth with epothilone a and b content of 550mg / L detected by HPLC through an organic membrane with a molecular weight cut-off of 6000 to obtain a total of 99ml of filtrate and lotion, and then pass the filtrate and lotion through ceramic Membrane filtration, the specification of the ceramic membrane is 1000 molecular weight cut-off, and the filtrate and lotion are 72ml in total, and the lotion is ordinary distilled water, and the collected filtrate and lotion are eluted through a SephadexG-15 gel chromatography column, and the eluent is: acetone It is prepared with petroleum ether at a volume ratio of 1:7. HPLC detects the eluent, collects the fractions whose total chromatographic purity of epothilone a and b reaches more than 80%, obtains 53ml of eluent, and then elutes Concentrate the solution through a nanofiltration membrane with a molecular weight cut-off of 500 to obtain 48ml of a concentrated solution. The concentrated solution is ...

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Abstract

The invention relates to a membrane extraction separation technique of epothilone, which comprises the following steps: filtering an epothilone a and b fermentation liquid through an organic membrane to obtain an organic membrane filtrate, and filtering the organic membrane filtrate through a ceramic membrane to obtain a ceramic membrane filtrate; eluting with a gel chromatographic column, sectionally collecting the eluates subjected to HPLC (high performance liquid chromatography) detection, concentrating the eluates through a nanofiltration membrane, eluting the concentrated solution by normal-phase silica gel chromatography, sectionally collecting the eluates subjected to HPLC detection, and crystallizing the collected eluates at low temperature to obtain the epothilone a and b crystal. The molecular weight cutoff of the organic membrane is 5000-6000. The molecular weight cutoff of the ceramic membrane is 1000-1500. The molecular weight cutoff of the nanofiltration membrane is 500-800. The crystallization temperature is -30 to -10 DEG C. The normal-phase chromatography is utilized to separate the epothilone a and b in the concentrated solution, thereby reducing the consumption of the organic solvent, simplifying the extraction process and lowering the cost.

Description

technical field [0001] The invention belongs to a method for filtering and extracting antitumor compounds, in particular to a membrane extraction and separation process for epothilone. Background technique [0002] Epothilone (epothilone) is a class of macrolide compounds with strong anti-tumor activity and can kill cancer cells at a lower concentration. The biosynthesis is mainly through the fermentation of myxobacteria to produce epothilone , biological fermentation mainly produces epothilone a and epothilone b, after research in related human cell lines, it was found that the anticancer effect of epothilone a is weaker than that of paclitaxel, while the anticancer effect of epothilone b is stronger than that of paclitaxel For paclitaxel, the yield is mainly measured by the titer of epothilone b at present. [0003] Biological fermentation mainly produces epothilone a and epothilone b, and its filtration is generally through resin analysis, but the expensive equipment, th...

Claims

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Application Information

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IPC IPC(8): C07D493/04
CPCC07D493/04
Inventor 虞龙李玉燕姚驰亚李市场楼干兵
Owner 虞龙
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