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Method for isolating and purifying ascomycin from streptomyces fermentation broth

A technology for purifying ascomycin and streptomyces, applied in organic chemistry and other directions, can solve the problems of affecting product purity and product form, high separation cost, low yield of ascomycin, etc., achieving easy industrial production, simple process, shortening The effect of extraction time

Inactive Publication Date: 2017-05-31
WUXI FORTUNE PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main problems in the separation and extraction of ascomycin at this stage are the low yield of ascomycin and the high cost of separation.
In addition, due to the isomerism of ascomycin in the water phase, the current extraction and separation methods mostly contain water phase, and the product contains ascomycin structural analogues, which affect the purity and shape of the product

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] A method for separating and purifying ascomycin from Streptomyces fermented liquid comprises the following steps.

[0030] Step 1), adding perlite equivalent to 1% of the volume of the Streptomyces fermentation broth as a filter aid to the Streptomyces fermentation broth, stirring for 30 minutes, separating solid and liquid, filtering, and collecting mycelia.

[0031] Step 2), soak and extract the mycelia with ethyl acetate at room temperature for 2 times, soak and extract for 2 hours each time to obtain the extract, combine the extract, and concentrate under reduced pressure under a negative pressure environment of 0.06MPa to obtain concentrated things.

[0032] Step 3), the concentrate obtained in step 2) is adsorbed with a bed filled with X-5 macroporous resin, and 20% vol acetone aqueous solution is added as an eluent for gradient elution, and the eluate is collected and concentrated.

[0033] Step 4), adding ethyl acetate to the concentrate obtained in step 3) for...

Embodiment 2

[0039] A method for separating and purifying ascomycin from Streptomyces fermented liquid comprises the following steps.

[0040] Step 1), adding diatomite equivalent to 4% of the volume of the Streptomyces fermentation broth as a filter aid to the Streptomyces fermentation broth, stirring for 40 minutes, solid-liquid separation, filtering, and collecting mycelia.

[0041] Step 2), soak and extract the mycelia with ethanol at room temperature for 5 times, each soaking and extracting for 6 hours, to obtain extracts, combine the extracts, and concentrate under reduced pressure under a negative pressure environment of 0.1 MPa to obtain a concentrate.

[0042] Step 3), the concentrate obtained in step 2) is adsorbed with a bed filled with D1300 macroporous resin, and gradient elution is performed with an aqueous solution containing 70% vol ethanol as the eluent, and the eluate is collected and concentrated.

[0043] Step 4), adding the concentrate obtained in step 3) to chloroform...

Embodiment 3

[0049] A method for separating and purifying ascomycin from Streptomyces fermented liquid comprises the following steps.

[0050] Step 1), adding a mixture of diatomite and perlite equivalent to 3% of the volume of the Streptomyces fermentation broth into the Streptomyces fermentation broth as a filter aid, stirring for 45 minutes, separating solid and liquid, filtering, and collecting Mycelium.

[0051] Step 2), extract the mycelium by immersing in acetone at room temperature for 3 times, each soaking and extracting for 3 hours, to obtain extracts, combine the extracts, and concentrate under reduced pressure under a negative pressure environment of 0.07MPa to obtain a concentrate.

[0052] Step 3), the concentrate obtained in step 2) is adsorbed with a bed filled with X-5 macroporous resin, and 55% vol of acetone aqueous solution is used as an eluent for gradient elution, and the eluate is collected and concentrated.

[0053] Step 4), the concentrate obtained in step 3) was ...

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PUM

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Abstract

The invention provides a method for separating and purifying ascomycin from streptomyces fermentation broth. The method comprises the following steps: filter-pressing the streptomyces fermentation broth with a plate frame to obtain thalli, extracting the thalli with an organic solvent at room temperature for multiple times, combining extracting solutions, concentrating the extracting solution and then transferring to macroporous resin and silica gel columns for chromatography, concentrating and crystallizing the obtained ascomycin with a mixed solvent, transferring the crystallized crude product to a dynamic axial high-pressure chromatographic column for isolation, and then concentrating and crystallizing the obtained ascomycin with the mixed solvent. In the invention, the prepared ascomycin has the purity of 98% or more and the yield of 80 to 90%. The method for separating and purifying ascomycin from the streptomyces fermentation broth, provided by the invention, has the advantages of greatly shortening the extraction time and improving the efficiency, having convenience in operation, simple process, low cost, easiness in industrial production and the like, and providing a feasible isolation process for the production of ascomycin.

Description

technical field [0001] The invention relates to the technical field of biopharmaceuticals, in particular to a method for separating and purifying ascomycin from streptomyces fermentation broth. Background technique [0002] Ascomycin (Ascomycin, Immunomycin, FK-520) is a three-carbon macrolide compound, and is an ethyl analog of the immunosuppressant FK-506 (tacrolimus, tacrolimus). Since the discovery of its immunosuppressive activity similar to tacrolimus, the research on the immunosuppressive activity of ascomycin and its derivatives has been rapidly promoted. Studies have found that John Ascomycin has a strong immunosuppressive effect, and currently Ascomycin has been used as an intermediate in the production of semi-synthetic APIs (such as pimecrolimus). Pimecrolimus is mainly used in the treatment of autoimmune diseases such as atopic dermatitis, allergic contact dermatitis, psoriasis, lupus erythematosus, lichen planus, vitiligo, and graft-versus-host disease. [00...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D498/18
CPCC07D498/18
Inventor 赵永俊付静曹峥杨亮游云龙蔡云洁金丹甜
Owner WUXI FORTUNE PHARMA
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