Strategy to overcome a nirmatrelvir resistance mechanism in the SARS-CoV-2 nsp5 protease.

Neilsen, Grace; Lan, Shuiyun; Slack, Ryan L; Lorson, Zachary C; Emanuelli Castaner, Andres; Lee, Rachel; Edwards, Kristin G; Zhang, Huanchun et al. · Sci Adv · 2025

basic_science · Level V

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Abstract

E166V in the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nsp5 protease confers strong resistance to the antiviral component of Paxlovid, nirmatrelvir (NIR), in passaging and clinical samples. In SARS-CoV-2 replicons, E166V drastically decreased Washington (WA1) but not Omicron (BA.1) fitness (20- versus 2-fold), suggesting a lower barrier to resistance in the BA.1 strain and consistent with observed differences in respective nsp5 dimerization affinities. Crystal structures reveal a steric clash between the rigid, bulky NIR <i>tert</i>-butyl group and the β-branched Val<sup>166</sup>, disrupting the covalent binding of NIR to the catalytic Cys<sup>145</sup> and leading to high resistance in BA.1 and WA1 replicons. NIR-resistant replicons remained susceptible to GC376, which can still covalently bind Cys<sup>145</sup> by avoiding a steric clash with Val<sup>166</sup> through "wiggling and jiggling." Hence, strategic flexibility is a strategy that will help design second-generation antivirals against NIR-resistant viruses.

Medical subject headings