Application of compound in preparation of drugs for resisting picornavirus
A technology of RNA virus and compound, which is applied in the field of preparation of anti-picorna virus drugs, and can solve the problems of poor anti-virus effect
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Embodiment 1
[0126] Embodiment 1 Drug Gradient Inhibition Experiment
[0127] RD (human malignant embryonic rhabdomyoma cells) cells were divided into 1×10 4 Cells / well were seeded in 96-well tissue culture plates (Corning, USA) and incubated at 37°C, 5% CO 2 Incubate in atmosphere for 3 days until approximately 100% confluency is reached. After forming a monolayer of RD cells at the bottom of the well, add 60 μL of diluted virus stock solution GZ203ACL21 with an appropriate titer to each well of the 96-well plate (2-8 plaques can be formed in the control well). Then place the plate at 37 °C, 5% CO 2 After incubation for 1 hour in the incubator, overlay medium was added directly to each well. Dilute 200 mM drug (dissolved in DMSO) with DMEM medium to 1 mM concentration. Then, 1.6 μL, 3.2 μL, 6.4 μL, 12.8 μL, 16 μL, and 25.6 μL of the compound were directly added to the covering medium of each well (3-6 duplicate wells). at 37°C, 5% CO 2 After culturing in the incubator for 2 days, pl...
Embodiment 2
[0135] Inhibition to EV-A71 virus in embodiment 2RD cells
[0136] RD cells at 1 x 10 5 cells / well were seeded in a 24-well plate. 37°C, CO 2 Incubate in the incubator until the cells cover the bottom of the plate. Set the titer to 1 x 10 4 PFU / ml of EV-A71 virus liquid GZ203AKL21 was added to each well in an amount of 200 μL per well. 37°C, CO 2 Incubate in an incubator for 1 hour, discard the virus solution in the well, wash each well twice with PBS, and then add 1ml of DMEM culture solution to each well. Then the candidate drug with a concentration of 1 mM was added to the wells in the amounts of 0 μL, 20 μL, 40 μL, 60 μL, 80 μL, and 100 μL, and three replicate wells were set for each concentration. 37°C, CO 2 After incubation in the incubator for 24 hours, 50 μL of the supernatant was added to the virus RNA extraction kit to start RNA extraction. A reverse transcription kit was used for RNA to generate cDNA, which was used as a template for quantitative PCR amplifi...
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