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Purification method for ascomycin

A technology of ascomycin and purification method, applied in the field of biopharmaceuticals, can solve the problems of rare extraction and purification of ascomycin, occupation, etc., and achieve the effects of easy industrial production, quality assurance, and simple preparation process

Inactive Publication Date: 2018-08-17
HISUN PHARMACEUTICAL (HANGZHOU) CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, the research on ascomycin in my country is still in its infancy, and most of the research work is concentrated on the strain selection and fermentation of ascomycin, the synthesis and development of ascomycin derivatives, antibacterial activity and clinical application, etc. There are few reports on the extraction and purification of ascomycin, and its market is basically occupied by foreign companies such as Japan and the United States

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  • Purification method for ascomycin
  • Purification method for ascomycin

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Experimental program
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Effect test

Embodiment 1

[0026] Get 6.0g ascomycin crude product (ascomycin chromatographic purity 67.2%, FK523 chromatographic purity 6.6%), dissolve with ethanol to 500mL to obtain sample solution, use the macroporous adsorption resin column that model is XAD-1600N to carry out dynamic adsorption, put The column flow rate is 0.5BV / h, the desorption mobile phase is 85% (mL / mL) ethanol aqueous solution, the elution flow rate is 1.0BV / h, and the liquid phase detection is used after every 0.5BV collection; Combined, concentrated under reduced pressure at 36°C until no liquid dripped out, then added 1L of ethyl acetate for extraction, then concentrated the ethyl acetate extract phase at 36°C under reduced pressure to obtain 3.7 g of dry powder. Dissolve the obtained dry powder with 46 mL of acetonitrile with a weight-to-volume ratio of about 8:100 (g / mL). After dissolving and clarifying, 49 mL of column liquid is obtained. The concentration of ascomycin detected by liquid phase is 58 mg / mL. The upper col...

Embodiment 2

[0028] Take 5.7g of ascomycin crude product (ascomycin chromatographic purity 64.7%, FK523 chromatographic purity 6.5%), dissolve it with ethanol to 500mL to obtain a sample solution, and use a model XAD-1600N macroporous adsorption resin column to carry out dynamic adsorption. The column flow rate is 0.5BV / h, the desorption mobile phase is 75% (mL / mL) ethanol aqueous solution, the elution flow rate is 1.0BV / h, and the liquid phase detection is used after every 0.5BV collection; Combined, concentrated under reduced pressure at 39°C until no liquid dripped out, then added 1L of ethyl acetate for extraction, then concentrated the ethyl acetate extract phase under reduced pressure at 39°C to obtain 3.8 g of dry powder. Dissolve the obtained dry powder with 63 mL of acetonitrile with a weight-to-volume ratio of about 6:100 (g / mL). After dissolving and clarifying, 65 mL of column liquid is obtained. The concentration of ascomycin detected by liquid phase is 44.6 mg / mL. The upper co...

Embodiment 3

[0030]Take 6.5g of ascomycin crude product (ascomycin chromatographic purity 67.6%, FK523 chromatographic purity 6.4%), dissolve with ethanol to 500mL to obtain sample solution, use the macroporous adsorption resin column that model is XAD-1600N to carry out dynamic adsorption, put The column flow rate is 0.5BV / h, the desorption mobile phase is 90% (mL / mL) ethanol aqueous solution, the elution flow rate is 1.0BV / h, and every 0.5BV is collected and detected by liquid phase; Combined, concentrated under reduced pressure at 39°C until no liquid dripped out, then added 1L of ethyl acetate for extraction, then concentrated the ethyl acetate extract phase at 39°C under reduced pressure to obtain 4.1 g of dry powder. Dissolve the obtained dry powder with 46 mL of acetonitrile with a weight-to-volume ratio of 9:100 (g / mL). After dissolving and clarifying, 48 mL of column liquid is obtained. The concentration of ascomycin detected by liquid phase is 66.3 mg / mL. The upper column liquid ...

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Abstract

The invention discloses a purification method for ascomycin. The method comprises the following steps: gathering ascomycin loading solution by using a macroreticular resin, collecting eluant and concentrating, adding an extraction agent to concentrated solution and extracting, and concentrating an organic phase to be dry powder; and dissolving the dry powder to be clear methyl alcohol or acetonitrile solution, and preparing a pure ascomycin product in high purity by using a liquid chromatograph. The preparation technology has the good removal effect to a key impurity FK523 in the ascomycin, and the prepared product has the high purity.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals, and in particular relates to a method for purifying ascomycin. Background technique [0002] Ascomycin (ascomycin, immunomycin, FK520, formula 1) is a 23-carbon macrolide compound, which was isolated from the culture medium of Streptomyces hygroscopicus in the 1960s. The cyclic lactone antibiotics, which are also the ethyl analogs of the immunosuppressant tacrolimus (FK-506), have also rapidly promoted the immunosuppressive activity of ascomycin since the immunosuppressive activity of tacrolimus was discovered Research. European patent EP0323865 describes the use of immunosuppressant ascomycin, and Chinese patent CN101036649 discloses the use of immunosuppressant ascomycin in the preparation of drugs for the treatment of autoimmune diabetes and skin rejection. It is also effective in the treatment of arthritis and psoriasis. Ascomycin not only has a very good effect in the treatment of autoim...

Claims

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Application Information

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IPC IPC(8): C07D498/18
CPCC07B2200/07C07D498/18
Inventor 朱进伟石磊潘剑张敏彭湘屏高祥郑玲辉滕云
Owner HISUN PHARMACEUTICAL (HANGZHOU) CO LTD
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