Structure-based phylogeny identifies avoralstat as a TMPRSS2 inhibitor that prevents SARS-CoV-2 infection in mice
Young Joo Sun, … , Alexander G. Bassuk, Vinit B. Mahajan
Young Joo Sun, … , Alexander G. Bassuk, Vinit B. Mahajan
Published April 12, 2021
Citation Information: J Clin Invest. 2021;131(10):e147973. https://doi.org/10.1172/JCI147973.
View: Text | PDF
Research Article Infectious disease Article has an altmetric score of 16

Structure-based phylogeny identifies avoralstat as a TMPRSS2 inhibitor that prevents SARS-CoV-2 infection in mice

Abstract

Drugs targeting host proteins can act prophylactically to reduce viral burden early in disease and limit morbidity, even with antivirals and vaccination. Transmembrane serine protease 2 (TMPRSS2) is a human protease required for SARS coronavirus 2 (SARS-CoV-2) viral entry and may represent such a target. We hypothesized that drugs selected from proteins related by their tertiary structure, rather than their primary structure, were likely to interact with TMPRSS2. We created a structure-based phylogenetic computational tool named 3DPhyloFold to systematically identify structurally similar serine proteases with known therapeutic inhibitors and demonstrated effective inhibition of SARS-CoV-2 infection in vitro and in vivo. Several candidate compounds, avoralstat, PCI-27483, antipain, and soybean trypsin inhibitor, inhibited TMPRSS2 in biochemical and cell infection assays. Avoralstat, a clinically tested kallikrein-related B1 inhibitor, inhibited SARS-CoV-2 entry and replication in human airway epithelial cells. In an in vivo proof of principle, avoralstat significantly reduced lung tissue titers and mitigated weight loss when administered prophylactically to mice susceptible to SARS-CoV-2, indicating its potential to be repositioned for coronavirus disease 2019 (COVID-19) prophylaxis in humans.

Authors

Young Joo Sun, Gabriel Velez, Dylan E. Parsons, Kun Li, Miguel E. Ortiz, Shaunik Sharma, Paul B. McCray Jr., Alexander G. Bassuk, Vinit B. Mahajan

×

Figure 5

Avoralstat inhibits viral entry directed by SARS-CoV-2 spike proteins.

Options: View larger image (or click on image) Download as PowerPoint
Avoralstat inhibits viral entry directed by SARS-CoV-2 spike proteins. (...
(A and B) HEK cells treated with camostat, avoralstat, PCI-27483, antipain, or SBTI 2 hours before TMPRSS2 transfection. TMPRSS2 reduced autoproteolysis (increased TMPRSS2-FL signal; 1-way ANOVA followed by Dunnett’s multiple-comparison test; *P < 0.0332, **P < 0.0021, ***P < 0.0002 compared with vehicle; n = 5 for each group except vehicle and camostat [n =14]). Calu-3-cells were treated with compounds and inoculated with pseudovirions harboring (C) VSV-G or (D) SARS-CoV-2-spike-protein (n = 6; fit to the Hill equation). Calu-3 cells were treated with (E) 100 μM or (F) indicated concentrations of each compound, then incubated with SARS-CoV-2. Viral gRNA was measured after 24 hours. Data represent mean ± SEM; n = 3. (E) Compounds reduced viral signal at 100 μM (1-way ANOVA followed by Tukey’s multiple-comparison test; mean ± SEM; n = 3; ****P < 0.0001 compared with vehicle). (F) Viral signal was reduced beginning from 100 nM (2-way ANOVA followed by Dunnett’s multiple-comparison test; mean ± SEM; n = 3; *P < 0.0332, ****P < 0.0001 compared with vehicle).