Application of Compound ps-341 in the Preparation of Picornaviridae Enterovirus Inhibitors
A PS-341, RNA virus technology, applied in antiviral agents, medical preparations containing active ingredients, pharmaceutical formulas, etc.
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Embodiment 1
[0021] The application of compound PS-341 in the preparation of Picornaviridae Enterovirus genus virus inhibitors, said compound PS-341 has a structure shown in formula (I):
[0022] .
[0023] The Enteroviruses of the family Picornaviridae are EV D68 virus and CV A16 virus.
Embodiment 2
[0024] Example 2. The effect of PS-341 on the viability of Hela cells and RD cells.
[0025] Prepare RD cells and Hela cells in a 96-well plate, adjust the concentration of the cell suspension, and divide into 96-well plates, 3000-10000 cells / well. The cell growth density is moderate, and the PS-341 drug concentration gradient is set to 0nM, 6.25nM, 12.5nM, 25nM, 50nM, 100nM, 200nM, 400nM. Each group is set with a drug control and a cell control. The drug control group is DMSO with a concentration of 1%. , the cell control group was not treated, and each group had 3 replicate wells. The cell viability of each treatment group was detected by the MTT cell viability assay method.
[0026] virus culture
[0027] Rhabdomyosarcoma cells (RD), which are susceptible to enterovirus D68, were selected as the inoculation objects. Inoculate RD cells in T75 culture flasks, when the cell density is 80%-90%, remove the cell culture medium, add DMEM, take an appropriate amount of virus sto...
Embodiment 3
[0048] Example 3. Effects of different concentrations of PS-341 on the replication of enterovirus EV D68.
[0049] This experiment was carried out in RD cells and Hela cells.
[0050] The specific implementation steps of cell recovery are the same as in Example 2.
[0051] cell culture
[0052] 1) Culture of RD cells
[0053] RD cells were cultured in DMEM liquid medium containing 10% FBS, 1% penicillin-streptomycin, and incubated in an incubator containing 5% CO2 at 37°C. After about 48 hours, digest with trypsin containing 0.25% EDTA. After about 3-5 minutes, add fresh medium with 10% FBS to terminate. Centrifuge at room temperature at 800 rpm for 5 minutes. Discard the supernatant. Take an appropriate amount of fresh medium to resuspend. One part was placed in a culture bottle to continue culturing, and the rest was transferred to a 96-well cell culture plate.
[0054] 2) Culture of HeLa cells
[0055] HeLa cells were cultured in DMEM liquid medium containing 10% FBS a...
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