Cervinomycin B1, B2, B3 and B4 and production method and application thereof

A technology of cervitromycin and antibiotics, applied in the field of medicine and biology, can solve the problem of difficult control of drug-resistant bacteria

Active Publication Date: 2019-02-12
MEDICINE & BIOENG INST OF CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] With the abuse of antibiotics, the resistance of pathogenic bacteria to antibiotics is gradually increasing, and the spread of drug-resistant genes in the same or different species of bacteria through horizontal transfer also makes the problem of drug-resistant bacteria more difficult. control

Method used

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  • Cervinomycin B1, B2, B3 and B4 and production method and application thereof
  • Cervinomycin B1, B2, B3 and B4 and production method and application thereof
  • Cervinomycin B1, B2, B3 and B4 and production method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Embodiment 1: Streptomyces CGMCC NO.16425 solid-state fermentation culture

[0065] (1) Preparation of fresh spore suspension: Streptomyces CPCC NO.204980 spore suspension frozen in a refrigerator at minus 70°C was melted and evenly spread and inoculated on 7-10 spore medium plates (medium composition: starch 20.0 g / L, soybean cake powder 20.0g / L, agar powder 15.0g / L, pH7.0; plate diameter 9.0cm, each plate is filled with about 20 ml of medium). When cultured at 28°C for 8-9 days, a layer of gray spores grew on the surface of the plate medium. Wash the spores with 8.0-10.0ml sterile water on each plate, and obtain a concentration of about 1×10 after shaking. 8 Spores / ml of spore suspension for inoculation of fermentation medium plates.

[0066] (2) Solid-state fermentation culture: evenly spread the spore suspension on the fermentation medium plate (substrate composition: cornstarch 10.0g / L, cottonseed cake powder 10.0g / L, threonine 10.0g / L, agar powder 15.0 g / L, pH ...

Embodiment 2

[0067] Embodiment 2: Separation and purification of cercomycin B component

[0068] For the separation and purification process of cercomycin B1, B2, B3, and B4 components, please refer to figure 2 .

[0069] (1) Ethyl acetate extraction :

[0070] Streptomyces CPCC NO.204980 solid-state fermentation culture (about 30L), adding an equal volume of ethyl acetate for extraction, soaking and extracting for 2 days each time, and extracting twice in total. The ethyl acetate extract was collected and evaporated in vacuo to obtain a brown ethyl acetate extract (about 21 g).

[0071] (2) Refined product of cervitromycin B component mixture (reversed-phase silica gel column for impurity removal) :

[0072] Redissolve the brown ethyl acetate extract in an appropriate amount of methanol (about 200ml), and sonicate to induce dissolution. According to the ratio of 1:1.5, weigh about 31g of reverse phase (ODS) silica gel chromatography column filler, add it to the methanol solution,...

Embodiment 3

[0096] Embodiment 3: Oxidation preparation of cercomycin B2 and B4 components

[0097] The components B2 and B4 of cercomycin are the oxidation products of components B1 and B3 of cercomycin, respectively. In embodiment 2, the yield of preparing cercomycin B2 and B4 components by separation and purification is relatively low, so the pure or semi-pure products of cercomycin B1 and B3 components obtained in embodiment 2 can be utilized. The chemical oxidation method is more efficient to prepare cercomycin B2 and B4 fractions. details as follows:

[0098] (1) Weigh 25mg (or 5mg) of cercomycin B1 (or B3) component, and dissolve it in 25.0ml (5.0ml) dichloromethane-methanol (volume ratio 1:1) by ultrasonic; weigh 25mg (or 5 mg) of silver oxide was added to the solution and stirred at room temperature.

[0099] (2) Take 100 μl of the supernatant of the reaction solution every 1 hour, and filter it with a 0.22 μm filter membrane to remove silver oxide; after removing the solvent i...

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Abstract

The invention relates to cervinomycin B1, B2, B3 and B4 and a production method and application thereof. Chemical structures of the cervinomycin B1, B2, B3 and B4 are as shown in formulae of (1), (2),(3) and (4). The production method comprises the following steps: through fermenting and culturing streptomycete of which a preservation number is CGMCC NO. 16425, harvesting a fermentation culture object, and performing extracting, separating and purifying to obtain the cervinomycin B1-B4 components. The cervinomycin B1-B4 components are expected as a lead compound for researching and developingan anti-bacterial infection drug.

Description

technical field [0001] The invention belongs to the field of medical biotechnology, in particular, relates to a kind of cervinomycin (cervinomycin) B1, B2, B3, B4 and its production method and application. Background technique [0002] With the abuse of antibiotics, the resistance of pathogenic bacteria to antibiotics is gradually increasing, and the spread of drug-resistant genes in the same or different species of bacteria through horizontal transfer also makes the problem of drug-resistant bacteria more difficult. control. According to data from my country's Bacterial Resistance Surveillance Network, from 2005 to 2016, although the clinical detection rate of some drug-resistant bacteria such as methicillin-resistant Staphylococcus aureus (MRSA) has been controlled to a certain extent, it still remains at 38%. %; however, the clinical detection rate of some drug-resistant bacteria such as imipenem-resistant Acinetobacter baumannii increased from 30% to more than 60%; among...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D498/14A61P31/04C12P17/18C12R1/465
CPCA61P31/04C07D498/14C12P17/188
Inventor 武临专胡笑文江冰娅胡辛欣胡晓敏李书芬余利岩游雪甫
Owner MEDICINE & BIOENG INST OF CHINESE ACAD OF MEDICAL SCI
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