Query String: REMDESIVIR
Remdesivir BDBM429505 med.21724, Compound 178
- Antiviral Activity Assay A549-hACE2 cells were seeded 1.5 × 105 cells/well in DMEM complete into 24-well plates (0.5 mL/well) and then incubated for 16 h at 37 °C and 5% CO2. Cells were pretreated with compound for 1 h prior to infection performed using a clinical isolate of SARS-CoV-2 (SARS-CoV-2, isolate USA-WA1/ 2020) from BEI Resources. When 2-fold serial dilutions of compound (0.15−20 μM; remdesivir: 10 μM) added to the same volume of SARS-CoV-2 (final MOI = 0.01), the mixture was added to the monolayer cells and incubated for 1 h at 37 °C and 5% CO2. Afterward, the mixture was removed and replaced with 0.5 mL of infection media and incubated at 37 °C and 5% CO2. After 48 h, supernatants were harvested and processed for RT-qPCR.
- CPE inhibition assay Briefly, Vero E6 ACE2 cells were cultured in 96-well flat-bottom plates at a density of 2 × 104 cells per well. Following infection of the cells with a 100 TCID50 of SARS-CoV-2, the plates were incubated on a rocker in 37 °C for 45 min for virus adsorption. The cells were then washed with DMEM, and the medium containing the test compounds at the desired concentration was added. Both the uninfected cells and the infected cells treated with 10 μM remdesivir were used as controls. The antiviral efficacy of test compounds was determined by the uptake and subsequent extraction of neutral red dye. After infection (68 h), cells were incubated with 0.034% neutral red dye for 3 h at 37 °C. Free dye was washed from the wells, and dye uptake was quantified using a microplate reader with absorbance at 540 nm. Absorbance values were expressed as percentages of uninfected control cells, and EC50 values of the test compounds were determined using Prism (GraphPad).