2. Academic Validation
  3. Optimizing the pharmacokinetics and selectivity of TMPRSS2 inhibitors

Optimizing the pharmacokinetics and selectivity of TMPRSS2 inhibitors

  • Eur J Med Chem. 2025 Mar 28:294:117579. doi: 10.1016/j.ejmech.2025.117579.
Sára Ferková 1,  Matthieu Lepage 1,  Antoine Désilets 1,  Kevin Assouvie 1,  Gabriel Lemieux 1,  Isabelle Brochu 1,  Ulrike Froehlich 1,  Alice Gravel-Trudeau 1,  Jules Vastra 1,  François Jean 2,  Philippe Sarret 1,  Richard Leduc 1,  Pierre-Luc Boudreault 3
Affiliations

Affiliations

  • 1 Department of Pharmacology-Physiology, Faculty of Medicine and Health Sciences, Institut de Pharmacologie de Sherbrooke (IPS), Université de Sherbrooke, Sherbrooke, Québec, Canada.
  • 2 Department of Microbiology and Immunology, Life Sciences Institute, University of British Columbia, Vancouver, British Columbia, Canada.
  • 3 Department of Pharmacology-Physiology, Faculty of Medicine and Health Sciences, Institut de Pharmacologie de Sherbrooke (IPS), Université de Sherbrooke, Sherbrooke, Québec, Canada. Electronic address: Pierre-Luc.Boudreault@USherbrooke.ca.
PMID: 40382841 DOI: 10.1016/j.ejmech.2025.117579
Abstract

Since 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has undergone significant genomic mutations, contributing to resistance against existing 2019 coronavirus disease (COVID-19) treatments. In a previous study, we identified N-0385, a potent host-directed inhibitor of transmembrane serine protease 2 (TMPRSS2), which has therapeutic efficacy towards SARS-CoV-2 Infection. However, in further evaluation of its preclinical druggability, N-0385 displayed unfavorable pharmacokinetic properties, including high bioavailability (99 %) following intranasal (IN) administration. This can lead to substantial systemic exposure and potential adverse effects due to off-target interactions. Here, we designed a library of peptidomimetic compounds with P3 site modifications on an optimized scaffold. We sought to maintain sub-nanomolar potency against TMPRSS2 (Kis < 2 nM), reduce pseudovirus Infection, while addressing the lack of selectivity and excessive lung uptake. Notably, inhibitor 9, which contains Asp at the P3 position, achieved a two-fold increase in TMPRSS2 inhibitory potency (Ki = 0.13 ± 0.03 nM), a >700-fold selectivity over Factor Xa (FXa), and showed superior selectivity against Other proteases (matriptase, transmembrane serine protease 6 (TMPRSS6), Thrombin, Furin, and tPA). Despite concerns about the role of FXa in the coagulation cascade, compound 9 had no impact on coagulation or thrombolysis 2 h after in vitro treatment. In the air-liquid interface (ALI) model of the lung epithelium, compound 9 displayed a 1.5-fold decrease in permeability compared to N-0385 and demonstrated sustained stability in lungs (11 h) and plasma (13 h). Taken together, our data demonstrate that continued optimization of this type of inhibitors will lead to improved therapeutics for the treatment of SARS-CoV-2 Infection by IN administration.

Keywords

Air-liquid interface (ALI) model; Factor xa; Intranasal administration; Peptidomimetic; SARS-CoV-2; Selectivity; TMPRSS2.

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