Figure 4. The acrylamide moiety is required for the antiviral activity of QL-XII-47.

A. Huh7 cells were pre-treated with 2 µM QL-XII-47 for 6 hours and then washed prior to DV2 infection (MOI of 1) (pre-treat), or were treated with 2 µM QL-XII-47 for 24 hours post-infection (post-treat). Infectious virus released to the supernatants at 24 hours post-infection was quantified by FFA. Representative data (mean ±standard deviation of experimental duplicates) out of n=2 independent experiments are shown.

B. Huh7 cells were pre-treated with 2 μM QL-XII-47 for 6 hours and then washed prior to transfection (pre-treat) or were treated with 2 µM QL-XII-47 for 12 hours starting at the time of transfection (post-treat) with a DV2(GVD) reporter replicon. Luciferase activity was quantified at 12 hours post-transfection and is plotted as a percentage of the DMSO-treated samples. Representative data (mean ± standard deviation of experimental duplicates) out of n=2 independent experiments are shown.

C. Structures of parent compound QL-XII-47 and QL-XII-47R, a derivative in which the acrylamide moiety is replaced with a non-reactive propyl amide group.

D. The concentration-dependent effects of QL-XII-47 and QL-XII-47R were assessed by infection of Huh7 cells with DV2 (MOI of 1) followed by addition of inhibitors. Viral yield was measured 24 hours later and is plotted as a percentage of the DMSO control. Representative data (mean ± standard deviation of experimental duplicates) from n≥2 independent experiments are shown.

E. Huh7 cells were infected with DV2 (MOI of 1), then treated with the indicated compounds. Infectious virus released to the supernatants at 24 hours post- infection was quantified by FFA. Representative data (mean ± standard deviation of experimental duplicates) out of n>2 independent experiments are shown. The chemical structure of each molecule is shown on the side of the graph.