Separation and purification method of impurity A or impurity B of amphotericin B

An amphotericin, separation and purification technology, applied in chemical instruments and methods, organic chemistry, preparation of sugar derivatives, etc., can solve the problems of poor stability of amphotericin B impurities, and achieve difficult separation and preparation, high separation efficiency, Fast separation effect

Pending Publication Date: 2022-04-12
浙江天台药业股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, there are few investigation and research data on the amphotericin B impurity, the stability of the amphotericin B impurity is relatively poor, and the separation has certain difficulties. If a general separation and preparation method applicable to the amphotericin B impurity can be developed, the amphotericin The pure product of B impurity can perfect the impurity profile of amphotericin B, which is of great significance for the research of amphotericin B impurity

Method used

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  • Separation and purification method of impurity A or impurity B of amphotericin B
  • Separation and purification method of impurity A or impurity B of amphotericin B
  • Separation and purification method of impurity A or impurity B of amphotericin B

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1: Separation and purification of amphotericin B impurity A

[0031] (1) Weigh 50 mg of amphotericin B, add 5 mL of NMP, 35 mL of acetonitrile and 0.1 mL of concentrated hydrochloric acid. Stir ultrasonically until it is completely dissolved, then add 5mL of 10g / L ammonium acetate solution and mix well before injecting.

[0032] (2) Dynamic axial compression preparation chromatographic conditions: mobile phase A is 0.02M citric acid aqueous solution (ammonia water adjusts pH to 4.7), B phase is acetonitrile; mobile phase A: mobile phase B=67:33; detection wavelength is 383nm And 303nm; the flow rate is 50mL / min; the injection volume is 200mL, the 50DAC dynamic axial compression column is an ODS column, and the particle size is 10um.

[0033] Run the elution method according to the above-mentioned chromatographic conditions, collect the target impurity (RT=166.47min) fraction, and measure the purity of each fraction by analytical liquid chromatography, and th...

Embodiment 2

[0036] Embodiment 2: Separation and purification of amphotericin B impurity B

[0037] (1) Weigh 50 mg of amphotericin B, add 5 mL of NMP, 35 mL of methanol and 0.1 mL of concentrated hydrochloric acid. Stir ultrasonically until it is completely dissolved, and then destroy it in the dark for 30 minutes, then add 5mL of 10g / L ammonium acetate solution, mix well and then inject the sample.

[0038] (2) Dynamic axial compression preparation chromatographic conditions: mobile phase A is 0.02M citric acid aqueous solution (ammonia water adjusts pH to 4.7), B phase is acetonitrile; mobile phase A: mobile phase B=65:35; detection wavelength is 383nm ; The flow rate is 50mL / min; the injection volume is 200mL, and the 50DAC dynamic axial compression column is an ODS column with a particle size of 10um.

[0039] Operate the elution method according to the above-mentioned preparative liquid chromatography conditions, collect target impurity (RT=42.75min) fractions, and measure the purit...

Embodiment 3

[0042] Embodiment 3: the pretreatment of amphotericin B impurity A and impurity B

[0043] (1) Weigh 2000 mg of amphotericin B, add 150 mL of NMP, 700 mL of acetonitrile and 0.1 mL of concentrated hydrochloric acid. Stir ultrasonically until completely dissolved, then add 150mL of 10g / L ammonium acetate solution and mix evenly before sample injection, prepare chromatographic conditions according to (2) dynamic axial compression in Example 1, (3) desalting and (4) 30 ℃ decompression distillation and other steps, it is found that increasing the sample processing capacity, and the storage time is relatively long, it will produce other impurities, and the content of the target impurity A and impurity B is less, which is more unfavorable for separation, so in the preferred embodiment ratio is processed.

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Abstract

The invention provides a method for separating and purifying an impurity A and an impurity B of amphotericin B. According to the method, a chromatogram is prepared mainly through dynamic axial compression, and pure products of the impurity A and the impurity B of the amphotericin B are obtained. The method specifically comprises the following steps: accelerating destruction under different conditions to obtain impurity A and B solutions of amphotericin B, eluting by taking 0.02 M citric acid aqueous solution / acetonitrile as a mobile phase, collecting target impurity fractions, and determining the purity of the target impurity fractions by using analytical high performance liquid chromatography, wherein the purity can reach more than 90%. And changing a mobile phase, carrying out re-elution and desalination by using water / acetonitrile, collecting a target fraction, and carrying out vacuum concentration to obtain a target product. The related impurities of the amphotericin B have the characteristics of unstable sample, high separation and preparation difficulty and the like, and the separation and purification method of the impurity A and the impurity B of the amphotericin B has the advantages of high separation efficiency, high separation speed, simplicity and convenience in operation and the like.

Description

technical field [0001] The invention belongs to the field of medicine separation and purification, and in particular relates to a method for separating and preparing amphotericin B impurity A and impurity B. Background technique [0002] The number of patients with fungal diseases has increased with the use of advanced medical treatments such as organ transplantation, immunosuppressive drugs, radiotherapy, and anticancer chemotherapy, as well as human immunodeficiency virus (HIV) infection. Amphotericin B (Amphotericin B) is a polyene antifungal drug. Its main mechanism of action is to selectively combine with ergosterol in the cell membrane to increase the permeability of the membrane, so that important substances are leaked out and toxic substances are infiltrated. Reduce the vitality of fungi and achieve antifungal effects. Amphotericin B plays an important role in clinical treatment because of its broad antibacterial spectrum and strong antifungal effect. [0003] ...

Claims

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Application Information

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IPC IPC(8): C07H17/08C07H1/06
Inventor郭一平党万利杨晓章张一乐覃俐
Owner浙江天台药业股份有限公司