Dengue Virus Inhibitors as Potential Broad-Spectrum Flavivirus Inhibitors

<b>Background.</b> Flaviviruses spread from endemic to non-endemic areas, causing illness in millions of people worldwide. The lack of effective therapies and the rapid expansion of flaviviral infections worldwide emphasize the importance of finding effective antivirals to treat such diseases. <b>Objectives.</b> To find out the potential broad-spectrum flavivirus inhibitors among previously reported inhibitors of DENV2/DENV4. <b>Methods.</b> The cytotoxicity of compounds was tested using WST-1 assay. The compounds were tested for their ability to inhibit the infection of DENV2, ZIKV, KUNV, and TBEV, and the most active compounds were also analyzed using the replicon-based assay. Interactions of one of the identified inhibitors with possible viral targets were studied using molecular dynamics simulations. <b>Results.</b> Two out of eight previously reported DENV2/DENV4 inhibitors demonstrated the ability to inhibit all studied viruses at low micromolar concentrations. Compound <b>C6</b> demonstrated the ability to inhibit both DENV2 and TBEV. Compounds <b>C1</b> (lycorine), <b>C3</b> (mycophenolic acid), and <b>C7</b> (vidarabine) were demonstrated as inhibitors of TBEV infection for the first time. <b>Conclusions.</b> Several compounds, previously described as inhibitors of DENV, are also able to inhibit other flaviviruses. This work is the first report on the anti-TBEV activity of lycorine (<b>C1</b>) and mycophenolic acid (<b>C3</b>), as well as vidarabine (<b>C7</b>). In addition, this is the first experimental confirmation of the antiviral activity of compound <b>C5</b> and the lack of detectable antiviral activity of compound <b>C8,</b> demonstrating the necessity of experimental verification of the computational predictions.