TMPRSS2 Inhibitor Discovery Facilitated through an <i>In Silico</i> and Biochemical Screening Platform
Amanda L. Peiffer, Julie M. Garlick, Yujin Wu, Jesse W. Wotring, Sahil Arora, Alexander S. Harmata, Daniel A. Bochar, Corey R. J. Stephenson, Matthew B. Soellner, Jonathan Z. Sexton, Charles L. Brooks, Anna K. Mapp
Abstract
The COVID-19 pandemic has highlighted the need for new antiviral approaches because many of the currently approved drugs have proven ineffective against mitigating SARS-CoV-2 infections. The host transmembrane serine protease TMPRSS2 is a promising antiviral target because it plays a role in priming the spike protein before viral entry occurs for the most virulent variants. Further, TMPRSS2 has no established physiological role, thereby increasing its attractiveness as a target for antiviral agents. Here, we utilize virtual screening to curate large libraries into a focused collection of potential inhibitors. Optimization of a recombinant expression and purification protocol for the TMPRSS2 peptidase domain facilitates subsequent biochemical screening and characterization of selected compounds from the curated collection in a kinetic assay. In doing so, we identify new noncovalent TMPRSS2 inhibitors that block SARS-CoV-2 infectivity in a cellular model. One such inhibitor, debrisoquine, has high ligand efficiency, and an initial structure-activity relationship study demonstrates that debrisoquine is a tractable hit compound for TMPRSS2.