Abstract
Dengue fever is an acute infectious disease caused by dengue virus (DENV) and transmitted by Aedes mosquitoes. There is no effective vaccine or antiviral drug available to date to prevent or treat dengue disease. Recently, RNA-dependent RNA polymerase (RdRp), a class of polymerases involved in the synthesis of complementary RNA strands using single-stranded RNA, has been proposed as a promising drug target. Hence, we screened new molecules against DENV RdRp using our previously constructed virtual screening method. Mol-5, [1,2,4]triazolo[1,5-a]pyrimidine derivative, was screened out from an antiviral compound library (~8000 molecules). Using biophysical methods, we confirmed the direct interactions between mol-5 and purified DENV RdRp protein. In luciferase assay, mol-5 inhibited NS5-RdRp activity with an IC50 value of 1.28 ± 0.2 μM. In the cell-based cytopathic effect (CPE) assay, mol-5 inhibited DENV2 infectivity with an EC50 value of 4.5 ± 0.08 μM. Mol-5 also potently inhibited DENV2 RNA replication as observed in immunofluorescence assay and qRT-PCR. Both the viral structural (E) and non-structural (NS1) proteins of DENV2 were dose-dependently decreased by treatment with mol-5 (2.5–10 μM). Mol-5 treatment suppressed DENV2-induced inflammation in host cells, but had no direct effect on host defense (JAK/STAT-signaling pathway). These results demonstrate that mol-5 could be a novel RdRp inhibitor amenable for further research and development.
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Acknowledgements
The National Natural Science Foundation of China (81603118, 81700854), the Pearl River Nova Program of Guangzhou (201806010119), the Natural Science Foundation (2017A030313717), and the “New Drug Creation and Development” Major scientific and technological projects of Guangdong Province (2019B020202002) supported this work.
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XGY, WYW, and SWL designed the research; YHW, WYW, SXG, and SJH performed the research; XGC, MZ, XYW, LL, and XDT contributed new reagents or analytical tools; XGY and SWL analyzed the data; XY and KSN wrote the paper.
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Wan, Yh., Wu, Wy., Guo, Sx. et al. [1,2,4]Triazolo[1,5-a]pyrimidine derivative (Mol-5) is a new NS5-RdRp inhibitor of DENV2 proliferation and DENV2-induced inflammation. Acta Pharmacol Sin 41, 706–718 (2020). https://doi.org/10.1038/s41401-019-0316-7
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DOI: https://doi.org/10.1038/s41401-019-0316-7
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