Preparation method of high-purity fidaxomicin

Abstract

The invention discloses a method for separating and purifying fidaxomicin from fidaxomicin fermented mycelia. The method comprises the following steps of first filtering and extracting the fidaxomicin fermentation liquid, diluting the extracted liquor with water, then decoloring, adsorbing and analyzing the extracted liquor through macroporous resin, and concentrating, extracting and drying the analyzed liquor to obtain a fidaxomicin crude product; and dissolving the fidaxomicin crude product in a polarity organic solvent, injecting polymer microsphere columns to carry out the chromatographic separation to obtain the high-purity fidaxomicin fine powder. By adopting the method, the interference of fermented secondary metabolite can be effectively eliminated, and the quality and the yield of the fidaxomicin can be improved. The entire process is simple and easy to operate; the prepared fidaxomicin can be used for drugs; and the purity of the prepared fidaxomicin is more than 95 percent, and the whole-coarse total yield is more than 45 percent. The used solvent is a conventional solvent, small in toxicity and applicable to the industrial production.

Description

technical field [0001] The invention relates to a method for separating and purifying antibiotics, in particular to a method for separating and purifying fidaxomicin from fermented mycelium of fidaxomicin. Background technique [0002] Fidaxomicin is a new type of macrolide antibiotic with an 18-membered ring structure produced by the fermentation of cystic bacteria, with the molecular formula C 52 h 74 Cl 2 o 18 , the structural formula is as follows: [0003] [0004] Fidaxomicin structural formula [0005] Fidaxomycin is a narrow-spectrum antibacterial drug, mainly against Gram-positive aerobic and anaerobic bacteria, Clostridium difficile, and has good effect on methicillin-resistant Staphylococcus aureus, Clostridium perfringens and Enterococcus. Antibacterial effect, but very weak antibacterial activity against Gram-negative bacteria. MIC of Fidaxomycin against Clostridium difficile 50 and MIC 90 Both are significantly lower than vancomycin or metronidazole,...

AI Technical Summary

Problems solved by technology

This method has the following disadvantages: (1) After the fermentation broth is filtered, the resin and mycelium coexist, and the screening is difficult, the resin is difficult to reuse, and the environmental protection treatment is difficult; (2) The eluate is directly obtained without decolorization and impurity removal. , contains more impurities, and causes great damage to the downstream purification medium; (3) reversed-phase silica column chromatography uses MeOH / H 2 O / AcOH three-phase elution system, in which acetic acid will not only destroy the bonded phase of the column itself and reduce the utilization rate of the reversed-phase column, but also has a higher boiling point of 118°C. When recovering solvent, water and methanol can be recovered completely, but acetic acid can only recover a part, so that the proportion of the recovered mixed solvent changes and cannot be reused, which increases the production cost; Reaction occurs or remains in the crystallization solution, resulting in a decrease in the purity of fidaxomicin powder

The disadvantage of this method is that multiple chromatographic columns are repeatedly used, the preparation method is too complicated and cumbersome, and the yield will be low after repeated chromatographic column separations. In addition, thin-layer preparative chromatography is used for enrichment, and the batch capacity is too small. Not suitable for large-scale production

WO2011146621A2 discloses a method for preparing high-purity Fidaxomycin. In this method, the preparation liquid is used to prepare Fidaxomicin crude product, and Fidaxomicin fine powder with a purity greater than 93% can be obtained. Although the steps of this method are simple, but The required equipment is relatively expensive, and the amount of fine powder prepared is small, so it is not suitable for industrial production

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