Nirmatrelvir-resistant SARS-CoV-2 variants with high fitness in an infectious cell culture system

Yuyong Zhou, Karen Anbro Gammeltoft, Line Abildgaard Ryberg, Long V. Pham, Helena Damtoft Tjørnelund, Alekxander Binderup, Carlos Rene Duarte Hernandez, Carlota Fernandez-Antunez, Anna Offersgaard, Ulrik Fahnøe, Günther Herbert Johannes Peters, Santseharay Ramirez, Jens Bukh, Judith Margarete Gottwein*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

106 Citations (Scopus)
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Abstract

The oral protease inhibitor nirmatrelvir is of key importance for prevention of severe coronavirus disease 2019 (COVID-19). To facilitate resistance monitoring, we studied severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) escape from nirmatrelvir in cell culture. Resistant variants harbored combinations of substitutions in the SARS-CoV-2 main protease (Mpro). Reverse genetics revealed that E166V and L50F + E166V conferred high resistance in infectious culture, replicon, and Mpro systems. While L50F, E166V, and L50F + E166V decreased replication and Mpro activity, L50F and L50F + E166V variants had high fitness in the infectious system. Naturally occurring L50F compensated for fitness cost of E166V and promoted viral escape. Molecular dynamics simulations revealed that E166V and L50F + E166V weakened nirmatrelvir-Mpro binding. Polymerase inhibitor remdesivir and monoclonal antibody bebtelovimab retained activity against nirmatrelvir-resistant variants, and combination with nirmatrelvir enhanced treatment efficacy compared to individual compounds. These findings have implications for monitoring and ensuring treatments with efficacy against SARS-CoV-2 and emerging sarbecoviruses.

Original languageEnglish
Article numbereadd7197
JournalScience Advances
Volume8
Issue number51
ISSN2375-2548
DOIs
Publication statusPublished - 2022

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