Novel antibacterial application of NPS-2143
A technology of NPS-2143 and its application, which is applied in the direction of antibacterial drugs, nitrile/isonitrile active ingredients, etc.
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Embodiment 1
[0011] Example 1. NPS-2143 can inhibit the growth of Staphylococcus aureus.
[0012] Pick a single colony of ATCC 33591 or ATCC 25923 or ATCC 6538 and inoculate it in 5 mL of MHB medium, and culture overnight at 37°C and 250 rpm. Transfer the overnight culture solution to 5 ml MHB medium at a ratio of 1:100, 37 °C, 250 rpm, 3-4 h (cultivate until OD 600 =0.5~0.6). Dilute the bacterial solution to 1×10 with MHB medium 5 CFU / mL.
[0013] Using the double dilution method for high-throughput screening, dilute the drug comparison into a series of concentrations (such as: 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25, 0.125, 0.0625 μg / mL) and Mix it with the same amount of bacterial dilution and place it in a 96-well plate. Make 3-4 duplicate holes for each concentration of drug, and use linezolid as a positive control, and set a blank control without drug. Bacteria were cultured in the incubator for 24 hours, and the OD of each well was detected 600 value, calculate the inhibition rate, ...
Embodiment 2
[0017] Example 2. Time-killing curves of different concentrations of NPS-2143.
[0018] A single colony of ATCC 33591 or ATCC 25923 was picked and inoculated in 5 mL of MHB medium, and cultured overnight at 37 °C and 250 rpm. Transfer the overnight culture solution to 5 mL MHB medium at a ratio of 1:100, 37 °C, 250 rpm, 3-4 h (cultivate to OD 600 =0.5~0.6). Dilute the bacterial solution to 1×10 with MHB medium 5 CFU / mL, take 3 mL of the diluted bacterial solution and add it to the test tube. Add the optimal compound with the final concentration of 0 MIC, 1 / 4 MIC, 1 / 2 MIC, 1 MIC, 2 MIC, 4 MIC to each test tube respectively, and culture for 0 h, 2 h, 4 h, 6 h, 8 h , 12 h, 24 h, 36 h, 48 h, and 72 h were sampled and measured, and the bacterial solutions obtained at different time points were diluted by 10 -3 、10 -6 、10 -9 times, apply the bacterial solution on the MHA, place it in a 37°C incubator and incubate for 24 hours, and the final colony count is expressed in log10 / ...
Embodiment 3
[0020] Example 3, NPS-2143 can inhibit the growth of Staphylococcus aureus persistent bacteria.
[0021] Ciprofloxacin was used to control the time to construct the persistent bacteria model. The preserved ATCC 33591 was cultured overnight in 5 mL of MHB medium at 37 °C and 250 rpm. Transfer the overnight culture solution to 10 mL of MHB medium at a ratio of 1:1000, add 3 μg / mL ciprofloxacin control group and blank control group, 37 ℃, 250 rpm, at 0 h, 2 h, 4 h, 6 h, 8 h, 10 h, 12 h, 24 h to measure OD 600 value, and the bacterial solution was diluted by 10 -3 、10 -6 、10 -9 times, the smeared bacteria solution was placed on the MHA, placed in a 37°C incubator for 24 hours and counted, with time as the abscissa and Log (CFU / mL) as the ordinate to draw a graph to confirm the time point for the establishment of the persistent bacteria model.
[0022] The preserved ATCC 33591 was cultured overnight in 5 mL of MHB medium at 37 °C and 250 rpm. The overnight culture solution wa...
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