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New cyclic lipopeptides locillomycin A, B and C and preparation method thereof

A technology of roximycin and cyclolipopeptide, which is applied in the direction of peptide preparation methods, microorganism-based methods, biochemical equipment and methods, etc.

Inactive Publication Date: 2014-01-22
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This structural formula is different from all cyclolipopeptide antibiotics with known structures so far, and does not belong to any of the above-mentioned surfactant family clusters, iturin family clusters, and ubiquitin family clusters. A, B, C have related reports with the same structure and activity

Method used

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  • New cyclic lipopeptides locillomycin A, B and C and preparation method thereof
  • New cyclic lipopeptides locillomycin A, B and C and preparation method thereof
  • New cyclic lipopeptides locillomycin A, B and C and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] : the fermentation of Roximycin A, B, C

[0078] Firstly, Bacillus subtilis Bs916 grown on LB (10% tryptone, 5% yeast powder, 5% sodium chloride) slant was inoculated into test tube culture medium containing 5mL LB liquid, and shaken at 37°C and 200r / min for 24h ; Then the culture medium was transferred to a 333ml / L shake flask, cultured with shaking at 28°C and 180r / min for 72h, and a total of 80L of fermentation broth was harvested.

Embodiment 2

[0079] : the extraction of Roximycin A, B, C

[0080] The fermentation broth obtained above was centrifuged to remove the bacteria, and the pH was adjusted to 2.8 with concentrated hydrochloric acid to precipitate overnight. Centrifuge at 8000g for 25min to obtain a precipitate, which is extracted with methanol, and the extract is subjected to 0.22μmol·L -1 After microporous membrane filtration, 200ml of crude extract was obtained.

Embodiment 3

[0081] : the separation of Roximycin A, B, C

[0082]The crude extract was diluted with deionized water into 30% methanol aqueous solution, the pH value was adjusted to 7.0, and then loaded on NH 2 Solid phase extraction column, use 50% methanol water solution, 100% methanol, 0.5% formic acid methanol solution, 1% formic acid methanol solution, 2% formic acid methanol solution gradient elution, in 2% formic acid methanol elution solution contains the target compound.

[0083] Adjust the pH value of methanol eluting solution containing 2% formic acid to 7.0, blow dry and concentrate with nitrogen, dilute with deionized water to 30% methanol aqueous solution, and then load the sample on a C18 solid phase extraction column; use 30% of 3 times the column volume , 40%, 50%, 60%, 70%, 80% methanol aqueous solution gradient elution, wherein in containing 50% and 60% methanol water eluent, obtain the pure product of Roximycin A, B, C composition , a purity of 94.4%, a total of 80mg....

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PUM

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Abstract

The invention relates to a group of new cyclic lipopeptides of locillomycins A, B and C and a preparation method thereof, a medicinal composition with active ingredients of locillomycins A, B and C, and an application of the locillomycins A, B and C in preparation of antifungal, antibacterial and antiviral medicaments. The locillomycins A, B and C are new cyclic lipopeptides separated from a culture of Bacillus subtilis Bs916 strains, and a result of an in vitro activity test proves that the locillomycins A, B and C have very strong inhibitory activity to pathogenic fungi, bacteria and viruses, are used as a new antifungal, antibacterial and antiviral medicament and have a good development prospect.

Description

technical field [0001] The present invention relates to a group of new cyclic lipopeptide antibiotics (cyclic lipopeptides): roxithromycin A, B, C and their preparation methods, pharmaceutical compositions with roxithromycin A, B, C as active ingredients and Application of roxithromycin A, B, and C in the preparation of antifungal, bacterial, and viral drugs. Background technique [0002] More than 10,000 antibiotics derived from microorganisms have been discovered so far, many of which have been widely used as medicines, pesticides and industrial additives. Cyclic lipopeptide antibiotics are a class of antibiotics discovered from Bacillus in the 1940s with a wide range of biological activities. product. Lipopeptide antibiotics generally have strong antifungal activity and hemolytic properties, have certain antibacterial and viral activities, and some even have antitumor activity, and have a wide range of application values ​​in medicine, pesticides and industry. Due to t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C12P21/02C07K1/36C07K1/34C07K1/30C07K1/20A61K38/08A61P31/04A61P31/10A61P31/12C12R1/125
CPCG01N2030/8818C07K7/06A61K38/00A61P31/00A61P31/04A61P31/10A61P31/12G01N30/02
Inventor 罗楚平陈志谊刘雪辉郭俊尧王晓宇刘邮洲
Owner JIANGSU ACAD OF AGRI SCI
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