Application of houttuyfonate analogue in preparation of streptococcus mutans growth and biofilm inhibitor

A technology of streptococcus mutans and houttuyniacin, applied in the field of medicine, can solve problems such as differences in the sensitivity of antibacterial drugs, research on antibacterial efficacy, etc., and achieve the effects of small solute molecular weight, low cytotoxicity and simple structure

Active Publication Date: 2021-07-23
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the sensitivity of aerobic microorganisms and facultative anaerobic or anaerobic microorganisms to specific antibacterial drugs is quite different (Dione N, Khelaifia S, Lagier J-C, Raoult D. The aerobic activity of metronidazole against anaerobic bacteria.2015 ;45:537-40.)
However, the antibacterial effect of neohouttuydin sodium on facultative anaerobic or anaerobic bacteria such as Streptococcus mutans has not been studied so far.

Method used

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  • Application of houttuyfonate analogue in preparation of streptococcus mutans growth and biofilm inhibitor
  • Application of houttuyfonate analogue in preparation of streptococcus mutans growth and biofilm inhibitor
  • Application of houttuyfonate analogue in preparation of streptococcus mutans growth and biofilm inhibitor

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preparation example Construction

[0033] The preparation method of inhibitor is as follows:

[0034] Step 1: Dissolve 2 mg of neohouttuydin sodium in 2 mL of 5% dimethyl sulfoxide solvent to obtain an inhibitor mother solution with a concentration of neohouttuydin sodium of 10 mg / mL;

[0035] Step 2: Gradiently dilute the mother liquor obtained in step 1 with 5% dimethyl sulfoxide; add it to the liquid medium (beef heart and brain infusion BHI), and the concentration of the inhibitor is 50 μg / mL, 100 μg / mL; The final concentration of methyl sulfoxide was 0.5% (v / v).

[0036] The inhibitor can inhibit the growth of Streptococcus mutans planktonic bacteria and the formation of biofilm, and can inhibit the number of viable bacteria in the Streptococcus mutans biofilm. The inhibitor can be used as an oral streptococcus mutans biofilm inhibitor; it can be used as an anti-caries drug; and the drug is prepared by using neohouttuydine sodium as an active substance and pharmaceutically acceptable auxiliary materials. ...

Embodiment 1

[0044] According to the above-mentioned aseptic operation procedure, inoculate the suspension of Streptococcus mutans (bacteria number: 32401) into BHI liquid medium containing different concentrations of inhibitors (100 μg / mL and 50 μg / mL) respectively, to determine the effect of inhibitors on Inhibitory activity of Streptococcus mutans biofilm formation.

[0045] According to the ratio of 1:100, the above-mentioned Streptococcus mutans suspension was inoculated into BHI liquid medium containing inhibitors at a final concentration of 100 μg / mL and 50 μg / mL, respectively, and incubated at 37 °C in facultative anaerobic (5% CO 2 ) for 24 hours, during which the OD600 value of each well was read every 30 minutes. And compare it with the control group, that is, the bacterial suspension containing no inhibitor and only 0.5% DMSO to determine whether the planktonic growth of Streptococcus mutans is affected.

[0046] figure 1 For the results of this example, it can be seen from t...

Embodiment 2

[0048] According to the above-mentioned aseptic operation procedure, inoculate the suspension of Streptococcus mutans (strain number: 32401) into BHI liquid medium containing different concentrations of inhibitors (100 μg / mL and 50 μg / mL) to determine the inhibitor Inhibitory activity against Streptococcus mutans biofilm formation.

[0049] According to the ratio of 1:100, the above-mentioned Streptococcus mutans suspension was inoculated into the BHI sucrose liquid medium containing the final concentration of 100 μg / mL and 50 μg / mL inhibitor respectively, and was incubated at 37 °C in facultative anaerobic (5% CO 2 ) for 24 hours at 37°C under facultative anaerobic (5% CO 2 ) for 24 hours to form a biofilm.

[0050] Then, carefully suck out the planktonic bacteria and supernatant in the 96-well plate, wash the biofilm cells at the bottom with pH = 7.4, 0.01M phosphate buffer (the buffer was purchased from Gibco, product number: C10010500BT), and wash the biofilm cells with 1...

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Abstract

The invention discloses application of a houttuyfonate analogue in preparation of a streptococcus mutans growth and biological membrane inhibitor. The preparation method of the inhibitor comprises the following steps: 1, dissolving sodium new houttuyfonate in a dimethyl sulfoxide solvent having the concentration of 5 percent to obtain an inhibitor mother solution with the sodium new houttuyfonate concentration of 10 mg/mL; 2, carrying out gradient dilution on the mother solution obtained in the step 1 by using dimethyl sulfoxide having the concentration of 5 percent; and adding the inhibitor into a liquid culture medium, wherein the concentration of the inhibitor is 50-100 [mu]g/mL, and the final concentration of the dimethyl sulfoxide is 0.5% (v/v). The inhibitor can inhibit streptococcus mutans growth and biofilm formation, has the advantages of small molecular weight, simple structure, stable property, good biocompatibility and the like, can remarkably inhibit main pathogenic bacteria of caries, and has the potential of serving as anti-caries drugs.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to the application of a houttuyfonate analogue in the preparation of growth and biofilm inhibitors of Streptococcus mutans. Background technique [0002] Caries is the most common dental disease in humans, which causes progressive destruction of tooth color, shape and texture, seriously affects oral pronunciation, chewing, language and other functions, and is the main cause of toothache and tooth loss (US Department of Health and Human Services . Oral Health in America: A Report of the Surgeon General. Rockville: National Institute of Dental and Craniofacial Research, National Institutes of Health, 2000: 308.). 60%-90% of children and most adults suffer from it (Petersen P E, Bourgeois D, Ogawa H, et al. The global burden of oral diseases and risks to oral health[J]. Bulletin of the World Health Organization, 2005, 83:661-669.). In view of its high incidence, wide distribution, a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/255A61P31/04A61P1/02
CPCA61K31/255A61P31/04A61P1/02
Inventor 李雨庆税钰森
Owner SICHUAN UNIV
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