Trypstatin as a Novel TMPRSS2 Inhibitor with Broad-Spectrum 1 Efficacy Against Corona and Influenza Viruses

Lawrenz, J.; Wettstein, L.; Rodriguez Alfonso, A.; Nchioua, R.; von Maltitz, P.; Albers, D.; Zech, F.; Vandeput, J.; Stevaert, A.; Fois, G.; Preising, N.; Schmierer, E.; Almeida-Hernandez, Y.; Petersen, M.; Staendker, L.; Wiese, S.; Braubach, P.; Frick, M.; Sanchez-Garcia, E.; Sauter, D.; Kirchhoff, F.; Naesens, L.; Muench, J.

doi:10.1101/2025.01.14.632953

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bioRxiv · 10.1101/2025.01.14.632953

Trypstatin as a Novel TMPRSS2 Inhibitor with Broad-Spectrum 1 Efficacy Against Corona and Influenza Viruses

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Abstract

Respiratory viruses, such as SARS-CoV-2 and influenza, exploit host proteases like TMPRSS2 for entry, making TMPRSS2 a prime antiviral target. Here, we report the identification and characterization of Trypstatin, a 61-amino acid Kunitz-type protease inhibitor derived from human hemofiltrate. Trypstatin inhibits TMPRSS2 and related proteases, with IC50 values in the nanomolar range, comparable to the small molecule inhibitor camostat mesylate. In vitro assays demonstrated that Trypstatin effectively blocks spike-driven entry of SARS-CoV-2, SARS-CoV-1, MERS-CoV, and hCoV-NL63, as well as hemagglutinin-mediated entry of influenza A and B viruses. In primary human airway epithelial cultures, Trypstatin significantly reduced SARS-CoV-2 replication and retained activity in the presence of airway mucus. In vivo, intranasal administration of Trypstatin to SARS-CoV-2-infected Syrian hamsters reduced viral titers and alleviated clinical symptoms. These findings highlight Trypstatins potential as a broad-spectrum antiviral agent against TMPRSS2-dependent respiratory viruses. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=182 SRC="FIGDIR/small/632953v1_ufig1.gif" ALT="Figure 1"> View larger version (25K): org.highwire.dtl.DTLVardef@1b90566org.highwire.dtl.DTLVardef@117293org.highwire.dtl.DTLVardef@1794959org.highwire.dtl.DTLVardef@1be9090_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Lawrenz, J., Wettstein, L., Rodriguez Alfonso, A., Nchioua, R., von Maltitz, P., Albers, D., Zech, F., Vandeput, J., Stevaert, A., Fois, G., Preising, N., Schmierer, E., Almeida-Hernandez, Y., Petersen, M., Staendker, L., Wiese, S., Braubach, P., Frick, M., Sanchez-Garcia, E., Sauter, D., Kirchhoff, F., Naesens, L., Muench, J.. 2025-01-15. Trypstatin as a Novel TMPRSS2 Inhibitor with Broad-Spectrum 1 Efficacy Against Corona and Influenza Viruses. https://doi.org/10.1101/2025.01.14.632953

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