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A kind of preparation method and purification method of ergosterol compound

An ergosterol and compound technology, applied in the field of preparation of ergosterol compounds, can solve the problems of complex preparation process, poor process controllability, slow production efficiency and the like, and achieve the effects of simple process, short production cycle and low cost

Active Publication Date: 2020-05-19
SHANGHAI INST OF PHARMA IND CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The present invention aims to overcome the complex preparation process, complicated operation, long production cycle, Harsh conditions, slow production efficiency, expensive raw materials, high cost, poor process controllability and other defects provide a preparation method and purification method for ergosterol compounds

Method used

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  • A kind of preparation method and purification method of ergosterol compound
  • A kind of preparation method and purification method of ergosterol compound
  • A kind of preparation method and purification method of ergosterol compound

Examples

Experimental program
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Effect test

Embodiment 1

[0042] The fermentation of embodiment 1 fungus:

[0043] Inoculate the Fusarium oxysporum strains preserved in glycerol tubes (20% glycerol) into a 250ml Erlenmeyer shaker flask containing 50ml of seed medium, and cultivate them with shaking at 220r / min at 30°C for 24h as the seed solution.

[0044] Seed medium: yeast extract 5g / L, glucose 20g / L, peptone 10g / L, experimental water, pH5.5.

[0045] Inoculate 1ml of the seed solution into a 750ml Erlenmeyer shaker flask (containing 200ml of fermentation medium), and cultivate for 168h at 30°C and 220r / min. A total of 30L fungal fermentation broth was enriched.

[0046] The fermentation medium is: glucose 30g / L, (NH 4 ) SO 4 20g / L, yeast extract 10g / L, MgSO 4 ·7H 2 O 1g / L, FeSO 4 ·7H 2 O 0.05g / L, experimental water, pH 5.5.

[0047] Preparation of Fermentation Crude Product:

[0048] After enrichment to obtain 30L of fungal fermentation broth, centrifuge at 4000r / min for 30min, discard the supernatant, add 15L of acetone ...

Embodiment 2

[0049] Example 2 Preparation of 5,8-(14), 22-triene-7-one, 3-hydroxyl, (3β, 22E) ergosterol

[0050] 1. Product separation and purification:

[0051] The acetone extraction component 4004-E obtained in Example 1 was dissolved with 40ml of methanol, and reversed-phase silica gel column chromatography (80g, ODS-C 18 ) (YMC company, batch number: 9955) was eluted with 0%, 20%, 40%, 60%, 80%, 100% methanol-water mixture successively, flow rate 10ml / min, each gradient was eluted with 1L eluent The 100% methanol eluate was concentrated to dryness under reduced pressure, stored at low temperature (4°C) for subsequent use, and the separation and purification process is as follows: figure 2 shown. The chromatographic analysis conditions of the HPLC of product are as follows: Chromatographic column: Ultimate AQ-C 18 (5μm) (14×150mm), detection wavelength: 210nm, mobile phase: A) methanol B) water, flow rate: 1ml / min, column temperature: 40°C, sample injection: 10μl, elution concentr...

Embodiment 3

[0060] Change the fermentation medium in Example 1 to: glucose 20g / L, (NH4)SO4 10g / L, yeast extract 5g / L, MgSO4 7H2O 0.5g / L experimental water to 1 liter, pH 5.5. , the rest of the culture conditions were consistent with Example 1, separated according to the separation steps of Example 2, and finally obtained a single compound 2.8 mg, HPLC purity was 93.5%, and its physical and chemical properties and nuclear magnetic resonance spectrum were the same as Example 2.

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Abstract

The invention discloses a preparation method of ergosterol compounds. The 5,8-(14),22-triene-7-one,3-hydroxyl,(3beta,22E) ergosterol compounds are represented by the formula (I). The preparation method comprises the following steps: inoculating a seed solution of Fusarium to carry out fermentation culture so as to obtain a fermentation liquid; wherein the fermentation culture lasts for 144 to 240 hours, and the fermentation culture medium comprises glucose (20 to 40 g / L), (NH4)SO4 (10 to 30 g / L), yeast extract (5 to 15 g / L), MgSO4.7H2O (0.5 to 2 g / L), and FeSO4.7H2O (0 to 1 g / L). The provided preparation method and purification method have the advantages that the technology is simple, the conditions are mild, the cost is low, the production period is short, massive production can be realized under artificially controlled conditions, and the industrialization is promoted. The provided purification method can obtain target compounds with a HPLC purity of 90% or more, and the purity is high.

Description

technical field [0001] The invention relates to a preparation method and a purification method of ergosterol compounds. Background technique [0002] Ergosterol and its derivatives have pharmacological effects such as antiviral and antiarrhythmic. 5,8-(14), 22-triene-7-one, 3-hydroxy, (3β, 22E) ergosterol, also known as H1-A, is a unique ergot of Cordyceps sinensis (Brek) Sacc, Cs The sterol derivative can inhibit activated human glomerular kidney diaphragm cells and improve type A immunoglobulin nephropathy (US005582828A). [0003] Cordyceps sinensis is a complex of subseapots and larval corpses formed by Cordyceps sinensis fungus of the family Ergotaceae parasitizing on the larvae of bat moths. It is one of the traditional precious Chinese medicinal materials in my country. At present, the preparation of H1-A is mainly to isolate its asexual generation strain from the Cs fruiting body, inoculate the culture medium for strain culture, and transplant it to fresh PDA inclin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P33/00C07J9/00C12R1/77
Inventor 朱宝泉杨丽鸳林军胡海峰周斌
Owner SHANGHAI INST OF PHARMA IND CO LTD