Discovery of a potent anti-Zika virus benzamide series targeting the viral protein NS4B

Discovery of a potent anti-Zika virus benzamide series targeting the viral protein NS4B

PLOS Pathogens

10.1371/journal.ppat.1013609

https://doi.org/10.1371/journal.ppat.1013609

Donghoon Chung, Yuka Otsuka, Eunjung Kim, Sultan Ullah, Nicole M. Kennedy, Khac Huy Ngo, Jinjoo Kang, Reece Tomlinson, Brian Alejandro, Koji Barnaby, Jeffrey Miller, Justin Shumate, Chwee Fang Teo, Yaw Bia Tan, Che Shin Chew, CongBao Kang, Dahai Luo, Louis Scampavia, Timothy P. Spicer, Thomas D. Bannister

03 April 2026

Chung, D., Otsuka, Y., Kim, E., Ullah, S., Kennedy, N. M., Ngo, K. H., Kang, J., Tomlinson, R., Alejandro, B., Barnaby, K., Miller, J., Shumate, J., Teo, C. F., Tan, Y. B., Chew, C. S., Kang, C., Luo, D., Scampavia, L., Spicer, T. P., & Bannister, T. D. (2026). Discovery of a potent anti-Zika virus benzamide series targeting the viral protein NS4B. PLOS Pathogens, 22(4), e1013609. https://doi.org/10.1371/journal.ppat.1013609

Zika virus (ZIKV), a member of the Flaviviridae family, causes significant public health concerns through congenital Zika syndrome and Guillain-Barré syndrome, yet no effective anti-ZIKV drugs or vaccines are available. To address this critical need, we conducted phenotypic, cytopathic effect-based high-throughput screening followed by medicinal chemistry optimization and discovered novel benzamide anti-ZIKV leads. Current best compounds demonstrated superior potency (EC50 values 40–400 nM, CC50 > 50 µM) compared to NITD-008, the most potent known anti-ZIKV agent. Time-of-addition assays, resistant virus selection studies, and biophysical binding experiments confirmed that NS4B interference constitutes the primary antiviral mechanism. Notably, resistance mutations mapped to the C-terminus of NS4B, distinct from other flavivirus NS4B inhibitors targeting dengue or yellow fever viruses, revealing novel insights into a critical function of the region. These findings establish NS4B as an Achilles’ heel for flaviviruses and support the development of pan-flavivirus therapeutics targeting this essential viral protein.

Attribution 4.0 International (CC BY 4.0)

This research / project is supported by the NIH - NIH grant
Grant Reference no. : U19AI171954

https://oar.a-star.edu.sg/communities-collections/articles/23112

1553-7374

Experimental Drug Development Centre

https://doi.org/10.1371/journal.ppat.1013609