A Suite of TMPRSS2 Assays for Screening Drug Repurposing Candidates as Potential Treatments of COVID-19

J. H. Shrimp, J. Janiszewski, C. Z. Chen, M. Xu, K. M. Wilson, S. C. Kales, P. E. Sanderson, P. Shinn, Z. Itkin, H. Guo, M. Shen, C. Klumpp-Thomas, S. G. Michael, W. Zheng, A. Simeonov, M. D. Hall,

10.1101/2022.02.04.479134

Research curated

Abstract

SARS-CoV-2 is the causative viral pathogen driving the COVID-19 pandemic that prompted an immediate global response to the development of vaccines and antiviral therapeutics. For antiviral therapeutics, drug repurposing allowed for rapid movement of existing clinical candidates and therapies into human clinical trials to be tested as COVID-19 therapies. One effective antiviral treatment strategy used early in symptom onset is to prevent viral entry. SARS-CoV-2 enters ACE2-expressing cells when the receptor-binding domain of the spike protein on the surface of SARS-CoV-2 binds to ACE2 followed by cleavage at two cut sites on the spike protein. TMPRSS2 has a protease domain capable of cleaving the two cut sites; therefore, a molecule capable of inhibiting the protease activity of TMPRSS2 could be a valuable antiviral therapy. Initially, we used a fluorogenic high-throughput screening assay for the biochemical screening of 6030 compounds in NCATS annotated libraries. Then, we developed an orthogonal biochemical assay that uses mass spectrometry detection of product formation to ensure that hits from the primary screen are not assay artifacts from the fluorescent detection of product formation. Finally, we assessed the hits from the biochemical screening in a cell-based SARS-CoV-2 pseudotyped particle entry assay. Of the six molecules advanced for further studies, two are approved drugs in Japan (camostat and nafamostat), two have entered clinical trials (PCI-27483 and otamixaban), while the other two molecules are peptidomimetic inhibitors of TMPRSS2 taken from the literature that have not advanced into clinical trials (compounds 92 and 114). This work demonstrates a suite of assays for the discovery and development of new inhibitors of TMPRSS2.

Source: BioRxiv

Related molecules

Name	Synonyms	Genes
Nafamostat	Nafamostat mesylate, p-Guanidinobenzoic acid ester with 6-hydroxy-2-naphthamidine
PCI-27483
Otamixaban
Transmembrane serine 2	TMPRSS2, Serine protease 10	TMPRSS2, PRSS10
Camostat	Camostat mesylate, Camostat mesilate, FOY-305

Related interactions

Target	Drug	Type	Result
Transmembrane serine 2	Nafamostat	In vitro	positive
Transmembrane serine 2	PCI-27483	In vitro	positive
Transmembrane serine 2	Otamixaban	In vitro	positive
Transmembrane serine 2	Camostat	In vitro	positive