Method used for high-efficient purification of pneumocandins B0

A high-efficiency, extraction technology, which is applied in the preparation method of peptides, chemical instruments and methods, organic chemistry, etc., can solve the problems affecting the quality of caspofungin acetate, etc., to improve the purity and quality of the product, and the process is simple and easy line, improve the effect of product quality

Active Publication Date: 2014-07-23
JIANGSU SENRAN CHEM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

C 0 Impurities can participate in subsequent reactions and seriously affect the quality of caspofungin acetate

Method used

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  • Method used for high-efficient purification of pneumocandins B0

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Experimental program
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Effect test

Embodiment 1

[0035] The 11 tons of fermented liquid are first transferred to the acidification tank, the pH value of the fermented liquid is adjusted to 2.0-4.0 with oxalic acid, and plate and frame filter press. To obtain 1200 kg of fungus residue, add 6 tons of 75% ethanol solution, stir for 6 hours, and plate and frame filter again to obtain 6 tons of extract.

[0036]Transfer the extract to a 1000L enamel kettle again, control the temperature ≤ 50°C, and vacuum ≤ -0.07MPa to concentrate in vacuo. During the concentration process, observe that the liquid becomes turbid, then sample and centrifuge, and send the clear liquid for inspection. Neomercontin B 0 When the content is 48mg / L, the concentration is stopped to obtain 50kg of the concentrated extract mixture; 200kg of diatomite with a specification of 100 mesh is added to the concentrated extract mixture to fully absorb, wrap the crystals, add 250kg of deionized water to stir, and centrifuge to obtain Mokandine B 0 260kg of centrif...

Embodiment 2

[0060] The 11 tons of fermented liquid are first transferred to the acidification tank, the pH value of the fermented liquid is adjusted to 2.0-4.0 with oxalic acid, and plate and frame filter press. To obtain 1200 kg of fungus residue, add 6 tons of 75% ethanol solution, stir for 6 hours, and plate and frame filter again to obtain 5.8 tons of extract.

[0061] Transfer the extract to a 1000L enamel kettle again, control the temperature ≤ 50°C, and vacuum ≤ -0.07MPa to concentrate in vacuo. During the concentration process, observe that the liquid becomes turbid, then sample and centrifuge, and send the clear liquid for inspection. Neomercontin B 0 When the content is 30mg / L, the concentration is stopped, and 46kg of the concentrated extract mixture is obtained; 260kg of diatomite with a specification of 100 mesh is added to the concentrated extract mixture to fully absorb, wrap the crystals, then add 306kg of deionized water to stir, and centrifuge to obtain Mokandine B 0 3...

Embodiment 3

[0070] The 11 tons of fermented liquid are first transferred to the acidification tank, and the pH value of the fermented liquid is adjusted to 2.0-4.0 with oxalic acid, and plate-and-frame filter press. To obtain 1200 kg of bacterial residue, add 6 tons of 75% ethanol solution, stir for 6 hours, and press filter again to obtain 6.2 tons of extract.

[0071] Transfer the extract to a 1000L enamel kettle again, control the temperature ≤ 50°C, and vacuum ≤ -0.07MPa to concentrate in vacuo. During the concentration process, observe that the material liquid becomes turbid, take a sample and centrifuge, and send the clear liquid for inspection. Neomercontin B 0 When the content is 62mg / L, the concentration is stopped, and 56kg of the concentrated extract mixture is obtained; 300kg of diatomite with a specification of 100 mesh is added to the concentrated extract mixture to fully absorb, wrap the crystals, then add 356kg of deionized water to stir, and centrifuge to obtain Moka...

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Abstract

The invention discloses a method used for high-efficient purification of pneumocandins B0. The method comprises following steps: (1) pH value of a fermentation broth containing pneumocandins B0 is adjusted to 2.0 to 4.0, the fermentation broth is filtered, and a pneumocandins B0 extract liquid is obtained via extraction; (2) the extract liquid is condensed, diatomite is added for crystal packaging, water is added, and an obtained mixture is stirred and is subjected to centrifugation; (3) a solid material obtained via centrifugation is dissolved in ethanol, active carbon is added for decolouring, and an obtained mixed material is filtered; (4) an obtained filtrate is condensed, chloroform is added, an obtained solution is delivered through a silica gel column, and a pneumocandins B0 eluant is collected; and (5) the pneumocandins B0 eluant is condensed to be dry, and crystallization is carried out in a multiphase solvent system so as to obtain pneumocandins B0. The method is capable of avoiding application of resin columns and reducing amount of solvents used for elution greatly; active carbon can be used for removing pigments effectively; silica gel column chromatography is capable of removing related substances C0; and extraction purity can be as high as 99%. The method is simple and convenient; cost is reduced greatly; and industrialized production can be easily realized.

Description

technical field [0001] The invention belongs to the field of medicinal chemistry, and in particular relates to a highly efficient purified pneumocidine B 0 Methods. Background technique [0002] Pneumocandins B 0 (referred to as PB0) is produced by the mold Glarea lozoyensis Secondary metabolites synthesized by fermentation. Glarea lozoyensis In addition to the target product, the fermentation product has C 0 Components such as PB0 also include other impurities such as a large amount of pigment, so the extraction and purification process of PB0 is more difficult. [0003] Chinese invention patent 200910133118.0 discloses the preparation of pneumocandine B 0 The method, the main steps are: a) centrifugal nemocontin B 0 Fermentation broth, take the mycelia, and extract neomocontin B with methanol 0 ; b) Evaporate the normethanol extract to dryness, and then use n-butanol to extract Neomocontin B 0 ; c) Evaporate the n-butanol extract to dryness, and then use 70-80% me...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/56C07K1/14
Inventor 王其龙朱海吴长军施成张致慧
Owner JIANGSU SENRAN CHEM
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