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Linking biochemical and cellular efficacy of MERS coronavirus main protease inhibitors

La, V. N. T.; Lahav, N.; Rodriguez, M.; Diaz-Tapia, R.; McGovern, B.; Benjamin, J.; Barr, H.; Kang, L.; Chodera, J. D.; Minh, D.

2026-02-21 biochemistry
10.64898/2026.02.20.707097 bioRxiv
Show abstract

Compounds that bind to the Middle East Respiratory Syndrome Coronavirus (MERS-CoV) main protease (MPro) often produce biphasic concentration-response curves (CRCs) in biochemical assays; low concentrations activate the enzyme and high concentrations inhibit it. This biphasic behavior complicates data analysis. Here, we compare three approaches to data analysis: fitting the Hill equation to the activation phase, fitting it to the inhibition phase, and fitting an enzyme kinetics model that incorporates dimerization and ligand binding to the complete CRC. In the latter case, cellular efficacy is predicted by extrapolating the model to high enzyme concentrations. For compounds in our drug lead series, all three procedures yield inhibitory concentrations that are correlated with live-virus antiviral assays. The latter procedure provides the most accurate forecast of cellular efficacy rank. These data analysis procedures may be valuable for antiviral drug discovery against MERS-CoV MPro and other enzymes with similar kinetics.

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