Plant-derived antiviral drugs as novel hepatitis B virus inhibitors: Cell culture and molecular docking study

Parvez, Mohammad K.; Tabish Rehman, Md.; Alam, Perwez; Al-Dosari, Mohammed S.; Alqasoumi, Saleh I.; Alajmi, Mohammed F.

The HepG2.2.15 cells were seeded in flat-bottom 96 well plates (0.5 Â 10 5 /well in triplicate) and incubated overnight. Based on the cytotoxicity results, and previously reported assays, the treatment dose of 10 lg/ml was chosen for all pure compounds in anti-HBV experiments. Lamivudine (2 lM) prepared in DMSO and RMPI, was included as positive control (Parvez et al., 2006) while RPMI with 0.1% DMSO served as untreated control. Next day, the old media were replaced with fresh media containing compounds and controls, and the culture was incubated for 5 days as described elsewhere . Culture supernatants were replaced with fresh dose every alternate day and samples collected on day 1, 3, and 5 were saved at À20°C. The viral HBsAg production in the culture supernatant was analyzed using Monolisa HBsAg ULTRA Elisa Kit (BioRad, USA) as per the manufacturer's instructions. All data were presented as % inhibition of HBsAg expression in relation to the untreated control. The experiment was repeated to confirm the reproducibility.