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[Preprint]. 2024 Apr 29:2024.04.29.591502. [Version 1] doi: 10.1101/2024.04.29.591502

Crystallographic fragment screening delivers diverse chemical scaffolds for Zika virus NS2B-NS3 protease inhibitor development

Xiaomin Ni, Andre Schutzer Godoy, Peter G Marples, Michael Fairhead, Blake H Balcomb, Matteo P Ferla, Charles WE Tomlinson, Siyi Wang, Charline Giroud, Jasmin Cara Aschenbrenner, Ryan M Lithgo, Max Winokan, Anu V Chandran, Warren Thompson, Mary-Ann Xavier, Eleanor P Williams, Martin Walsh, Daren Fearon, Lizbé Koekemoer, Frank von Delft
PMCID: PMC11092485  PMID: 38746305

Abstract

Zika virus (ZIKV) infections cause microcephaly in new-borns and Guillain-Barre syndrome in adults raising a significant global public health concern, yet no vaccines or antiviral drugs have been developed to prevent or treat ZIKV infections. The viral protease NS3 and its co-factor NS2B are essential for the cleavage of the Zika polyprotein precursor into individual structural and non-structural proteins and is therefore an attractive drug target. Generation of a robust crystal system of co-expressed NS2B-NS3 protease has enabled us to perform a crystallographic fragment screening campaign with 1076 fragments. 48 binders with diverse chemical scaffolds were identified in the active site of the protease, with another 6 fragment hits observed in a potential allosteric binding site. Our work provides potential starting points for the development of potent NS2B-NS3 protease inhibitors. Furthermore, we have structurally characterized a potential allosteric binding pocket, identifying opportunities for allosteric inhibitor development.

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