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Method for purifying caspofungin precursor pneumocandin B0 component

A purification method, the technology of caspofungin, which is applied in the field of purification and synthesis of caspofungin, can solve the problems of unsatisfactory purification effect, difficulty in large-scale production, and reduced product yield, and achieve low cost and high equipment efficiency. The effect of low requirements and high production feasibility

Active Publication Date: 2012-12-12
CHENGDU YATU BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Currently, pneumocandinB has been 0 Reports on component purification, but the existence or purification effect is not ideal, or the operation is extremely complicated, resulting in a decrease in product yield, making it difficult to achieve large-scale production

Method used

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  • Method for purifying caspofungin precursor pneumocandin B0 component

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] 5g PB 0 The crude components were dissolved in 500ml of 45% ethanol (V / V), which also contained 1‰ formic acid (V / V), and then the pH value was adjusted to 3.0 with formic acid, and 500ml of filtrate was obtained by filtration, and the filtrate was reversed HPLC analysis, the main peak (including PB 0 , PC 0 Components, the two show the same chromatographic peak in reversed-phase chromatography) The content of the component is 3.62g, normal phase HPLC analysis (PB 0 , PC 0 Components can be separated), PB 0 The content of the components was 2.43 g, and the above filtrate was applied to a reverse phase column.

[0030] C18 reversed-phase column material (Fuji Corporation, Japan) Specifications: Lot.No.HU00200, Pro. No.SMB100, Size: 20 / 45um.

[0031] Weigh 300g of C18 reversed-phase column material, load the column with 95% ethanol (V / V) solution, and then equilibrate the column with 45% ethanol, containing 1‰ formic acid ((V / V) solution. After loading, use 50% ethan...

Embodiment 2

[0037] 5g PB 0 The crude component was dissolved in 500ml of 45% ethanol (V / V) solution, then adjusted to pH 4.0 with formic acid, and filtered to obtain 500ml of filtrate, which was analyzed by reverse phase HPLC, the main peak (containing PB 0 , PC 0 Components, the two show the same chromatographic peak in reversed-phase chromatography) The content of the component is 3.62g, normal phase HPLC analysis (PB 0 , PC 0 Components can be separated), PB 0 The content of the components was 2.43 g, and the above filtrate was applied to a reverse phase column.

[0038] C18 reversed-phase column material (Fuji Corporation, Japan) Specifications: Lot.No.HU00200, Pro. No.SMB100, Size: 20 / 45um.

[0039]Weigh 300g of C18 reverse-phase column material, pack the column with 95% ethanol solution, and then equilibrate the column with 45% ethanol containing 1‰ formic acid ((V / V) solution. After loading the sample, use 50% ethanol (V / V) (Which contains 1‰ formic acid (V / V)) to wash the col...

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Abstract

The invention relates to a method for purifying a caspofungin precursor pneumocandin B0 component, and the method comprises the following steps of: carrying out reversed-phase column chromatography on a PB0 component crude product, collecting and carrying out reduced-pressure concentration to obtain a semi-pure product; carrying out normal phase column chromatography on the PB0 semi-pure product obtained in the step 1 to obtain high-purity PB0 component; and carrying out positive and negative phase liquid phase analysis, wherein the chromatographic purity is more than 95%. Compared with the prior art, the method has the beneficial effects that a normal phase column and a reversed-phase column are used for purifying the PB0 component, so that the purification steps are simple, the purification effect is good, the requirements for equipment are low, large-scale production is realized, and the production feasibility is high; and particularly, the PB0 component separated by the method is high in yield, but is lower in cost.

Description

technical field [0001] The present invention relates to the synthesis and purification of caspofungin, in particular to the purification method of caspofungin precursor. [0002] Background technique [0003] Caspofungin (caspofungin) is the first new type of echinocandin antifungal drug approved for marketing. It is a semi-synthetic cyclic lipid compound with a relative molecular weight of 1213.42. Its precursor compound pneumocandinB 0 (PB 0 ) is extracted from Glarea lozoyensis. The target of caspofungin is the glucan synthase in the fungal cell wall, which is different from the traditional antifungal drugs that act on ergosterol in the fungal plasma membrane. Compared with the current three types of systemic antifungal drugs: polyenes (amphotericin B), pyrroles (such as ketoconazole, fluconazole, itraconazole), and fluoropyrimidines, it shows a wider range of antifungal agents. Antibacterial spectrum: It is effective against most Candida and other antifungal drug-re...

Claims

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Application Information

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IPC IPC(8): C07K7/06C07K1/20C07K1/16
Inventor 朱辉张翠英邹敬源朱春燕龙燕黄运昌杨益陈茂林杨旭成
Owner CHENGDU YATU BIOLOGICAL TECH
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