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Preparation method and application of quinoxaline amide compound used as streptococcus mutans biological membrane inhibitor

A technology of Streptococcus mutans and quinoxaline imine, which is applied in the digestive system, antibacterial drugs, drug combinations, etc., can solve the problem that fluoride cannot be targeted, and inhibit the formation and maturation of Streptococcus mutans biofilm , simple structure, small molecular weight effect

Active Publication Date: 2015-05-20
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The purpose of the present invention is to measure the inhibitory action of a kind of quinoxaline imine compound to streptococcus mutans biofilm, in order to provide a kind of quinoxaline imine compound as the streptococcus mutans biofilm inhibitor in suppressing this biofilm The preparation method and its application in the process of formation and maturation overcome the deficiency that the existing anti-caries drug fluoride cannot target the plaque biofilm

Method used

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  • Preparation method and application of quinoxaline amide compound used as streptococcus mutans biological membrane inhibitor
  • Preparation method and application of quinoxaline amide compound used as streptococcus mutans biological membrane inhibitor
  • Preparation method and application of quinoxaline amide compound used as streptococcus mutans biological membrane inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1: the preparation method of the culture of bacterial strain and liquid inhibitor

[0042] Preparation of test strains:

[0043] The test strain in this example involves Streptococcus mutans (strain number: 32401; obtained from China Medical Bacteria Collection and Management Center, website: http: / / www.cmccb.org.cn / ).

[0044] Components and preparation of medium:

[0045] Components of bovine heart and brain infusion (for convenience of description, hereinafter referred to as BHI) liquid medium and its preparation method: 37g of commercially available BHI powder (the BHI powder was purchased from Sigma, USA, item number: 53286) was added to 1000mL of distilled water Medium, high temperature and high pressure (121.3°C, 103.4kPa) sterilization for 15 minutes, cooled for later use. This liquid medium is a typical liquid medium for planktonic culture of Streptococcus mutans.

[0046] Components of BHI sucrose liquid medium and its preparation method: 37 g of...

Embodiment 2

[0052] Embodiment 2: Inhibition experiment of inhibitor to mutant streptococcus biofilm formation

[0053] According to the aseptic procedure of Example 1, the Streptococcus mutans suspension (bacteria number: 32401) of Example 1 was inoculated respectively in BHI sucrose liquid medium containing different concentrations of inhibitors, to determine the inhibitory effect on Inhibitory activity of Streptococcus mutans biofilm formation. The specific method is: according to the ratio of 1:100, the Streptococcus mutans suspension in Example 1 was respectively inoculated into BHI with concentration gradients of 0.05 μg / mL, 0.5 μg / mL, 5 μg / mL and 50 μg / mL inhibitors. In sucrose liquid medium, at 37 °C, facultative anaerobic (5% CO 2 ) for 16 hours. Then, carefully suck out the planktonic bacteria and supernatant in the 48-well plate, and use pH=7.4, 0.01M phosphate buffer (the buffer was purchased from Thermo Company, USA, product number: P5368). Dissolve one bag of content in 10...

Embodiment 3

[0054] Embodiment 3: Inhibitor experiments on the inhibition of Streptococcus mutans biofilm maturation

[0055] According to the aseptic operation procedure of Example 1, first inoculate the Streptococcus mutans suspension (bacteria number: 32401) of Example 1 in BHI sucrose liquid medium to form a biofilm, and then add inhibitors of different concentrations respectively To determine the inhibitory activity of inhibitors on Streptococcus mutans biofilm maturation. The specific method is: inoculate the Streptococcus mutans suspension in Example 1 into a 48-well plate containing BHI sucrose liquid medium according to a dilution ratio of 1:100, and inoculate it at 37° C. under facultative anaerobic (5% CO 2 ) for 24 hours to form a biofilm. At this point, add inhibitors at final concentrations of 5 μg / mL and 50 μg / mL to the biofilm cultures of Streptococcus mutans, respectively, and incubate at 37°C in facultative anaerobic (5% CO 2) to continue culturing for 48 hours. Then, ...

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Abstract

The invention discloses a preparation method and application of a quinoxaline amide compound used as a streptococcus mutans biological membrane inhibitor and relates to the technical field of bacteriostatic drug screening. The inhibitor consists of compounds 2-(4-methoxy-phenyl)-N-(3-{[2-(4-methoxy-phenyl) ethyl] imido}-1,4-dihydro-2-quinoxaline subunit) ethylamine and dimethyl sulfoxide (DMSO). The preparation method comprises the following steps: (1) dissolving 50mg of powdery 2-(4-methoxy-phenyl)-N-(3-{[2-(4-methoxy-phenyl) ethyl] imido}-1,4-dihydro-2-quinoxaline subunit) ethylamine in 1mL of a dimethyl sulfoxide (DMSO) solvent to prepare an inhibitor mother solution of which the initial concentration is 50mg / mL; (2) performing gradient dilution on the inhibitor mother solution by using dimethyl sulfoxide (DMSO), and adding into a brain heart infusion (BHI) liquid culture medium to make sure that the final concentrations of the inhibitor are respectively 0.05mu g / mL, 0.5mu g / mL, 5mu g / mL and 50mu g / mL and the final concentrations of the solvent DMSO are all 1% (v / v). Application tests show that the inhibitor can be used as an oral mouth streptococcus mutans biological membrane inhibitor and a candidate targeted drug for a novel anti-caries strategy.

Description

technical field [0001] The invention belongs to the technical field of bacteriostatic drug screening, and in particular relates to a small molecular compound capable of inhibiting the formation and maturation of Streptococcus mutans biofilm, namely a preparation method of a quinoxaline imine compound as a Streptococcus mutans biofilm inhibitor and its preparation method. application. Background technique [0002] Caries is the most common oral disease in humans. It is a disease in which the hard tissue of the tooth is progressively destroyed under the action of various factors, mainly bacteria, and is manifested by changes in tooth color, shape, and texture. Caries seriously affects the pronunciation, chewing, language and other functions of the oral cavity. In severe cases, it can also lead to various diseases such as pulpitis and jaw osteomyelitis. Everyone is at risk of caries, whether they are infants, adolescents or adults. Dental caries has become a common oral healt...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/498A61P31/04A61P1/02
Inventor 李雨庆任智陈卢璐李明云李继遥邹静
Owner SICHUAN UNIV
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