TY - JOUR
T1 - A novel class of TMPRSS2 inhibitors potently block SARS-CoV-2 and MERS-CoV viral entry and protect human epithelial lung cells
AU - Mahoney, Matthew
AU - Damalanka, Vishnu C.
AU - Tartell, Michael A.
AU - Chung, Dong hee
AU - Lourenço, Andre Luiz
AU - Pwee, Dustin
AU - Mayer Bridwell, Anne E.
AU - Hoffmann, Markus
AU - Voss, Jorine
AU - Karmakar, Partha
AU - Azouz, Nurit P.
AU - Klingler, Andrea M.
AU - Rothlauf, Paul W.
AU - Thompson, Cassandra E.
AU - Lee, Melody
AU - Klampfer, Lidija
AU - Stallings, Christina L.
AU - Rothenberg, Marc E.
AU - Pohlmann, Stefan
AU - Whelan, Sean P.J.
AU - O’Donoghue, Anthony J.
AU - Craik, Charles S.
AU - Janetka, James W.
N1 - Funding Information:
ACKNOWLEDGMENTS We thank David Griggs and Scott Campbell at Saint Louis University for conducting the pharmacokinetic studies on MM3122. We also thank Michael Winter at University of California, San Francisco for his help with the initial MSP-MS and PS-SCL profiling. This work was funded by grants from the Siteman Cancer Center (Grants 16-FY18-02 and SCC P30CA091842) of Washington University and Barnes Jewish Hospital Foundation (Grant BJHF 4984) in Saint Louis (J.W.J.). This work was also funded by NIH Awards R43 CA243941 (J.W.J. and L.K.), R43 CA224832 (J.W.J. and L.K.), U19 AI142784 (C.L.S.), P50AI150476 (C.S.C.), and U19 AI070235 (M.E.R.), and by the Campaign Urging Research for Eosinophilic Diseases Foundation (M.E.R.). Additional support was provided by a Fast Grant from Emergent Ventures at the Mercatus 9 Center, George Mason University (C.S.C.). S.P. acknowledges funding by Bundesministerium fu€r Bildung und Forschung (German: Federal Ministry of Education and Research; Bonn, Germany); RAPID Consortium, awards 01KI1723D and 01KI2006D; RENACO award 01KI20328A; SARS_S1S2 award 01KI20396; Charité – Universita€tsmedizin Berlin (For-schungsnetzwerk der Universita€tsmedizin zu Covid, COVIM consortium, award 01KX2021); the country of Lower Saxony (grant 14-76103-184); and the German Research Foundation (grants PO 716/11-1 and PO 716/14-1). Work with live SARS-CoV-2 was funded by a Burroughs Wellcome Fund Investigators in the Pathogenesis of Infectious Disease Award (17008, C.L.S.).
Funding Information:
We thank David Griggs and Scott Campbell at Saint Louis University for conducting the pharmacokinetic studies on MM3122. We also thank Michael Winter at University of California, San Francisco for his help with the initial MSP-MS and PS-SCL profiling. This work was funded by grants from the Siteman Cancer Center (Grants 16-FY18-02 and SCC P30CA091842) of Washington University and Barnes Jewish Hospital Foundation (Grant BJHF 4984) in Saint Louis (J.W.J.). This work was also funded by NIH Awards R43 CA243941 (J.W.J. and L.K.), R43 CA224832 (J.W.J. and L.K.), U19 AI142784 (C.L.S.), P50AI150476 (C.S.C.), and U19 AI070235 (M.E.R.), and by the Campaign Urging Research for Eosinophilic Diseases Foundation (M.E.R.). Additional support was provided by a Fast Grant from Emergent Ventures at the Mercatus 9 Center, George Mason University (C.S.C.). S.P. acknowledges funding by Bundesministerium fur ? Bildung und Forschung (German: Federal Ministry of Education and Research; Bonn, Germany); RAPID Consortium, awards 01KI1723D and 01KI2006D; RENACO award 01KI20328A; SARS_S1S2 award 01KI20396; Charite ? Universitatsmedizin ? Berlin (Forschungsnetzwerk der Universitatsmedizin ? zu Covid, COVIM consortium, award 01KX2021); the country of Lower Saxony (grant 14-76103-184); and the German Research Foundation (grants PO 716/11-1 and PO 716/14-1). Work with live SARS-CoV-2 was funded by a Burroughs Wellcome Fund Investigators in the Pathogenesis of Infectious Disease Award (17008, C.L.S.).
Publisher Copyright:
© 2021 National Academy of Sciences. All rights reserved.
PY - 2021/10/26
Y1 - 2021/10/26
N2 - The host cell serine protease TMPRSS2 is an attractive therapeutic target for COVID-19 drug discovery. This protease activates the Spike protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and of other coronaviruses and is essential for viral spread in the lung. Utilizing rational structure-based drug design (SBDD) coupled to substrate specificity screening of TMPRSS2, we have discovered covalent small-molecule ketobenzothiazole (kbt) TMPRSS2 inhibitors which are structurally distinct from and have significantly improved activity over the existing known inhibitors Camostat and Nafamostat. Lead compound MM3122 (4) has an IC50 (half-maximal inhibitory concentration) of 340 pM against recombinant full-length TMPRSS2 protein, an EC50 (half-maximal effective concentration) of 430 pM in blocking host cell entry into Calu-3 human lung epithelial cells of a newly developed VSV-SARS-CoV-2 chimeric virus, and an EC50 of 74 nM in inhibiting cytopathic effects induced by SARS-CoV-2 virus in Calu-3 cells. Further, MM3122 blocks Middle East respiratory syndrome coronavirus (MERS-CoV) cell entry with an EC50 of 870 pM. MM3122 has excellent metabolic stability, safety, and pharmacokinetics in mice, with a half-life of 8.6 h in plasma and 7.5 h in lung tissue, making it suitable for in vivo efficacy evaluation and a promising drug candidate for COVID-19 treatment.
AB - The host cell serine protease TMPRSS2 is an attractive therapeutic target for COVID-19 drug discovery. This protease activates the Spike protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and of other coronaviruses and is essential for viral spread in the lung. Utilizing rational structure-based drug design (SBDD) coupled to substrate specificity screening of TMPRSS2, we have discovered covalent small-molecule ketobenzothiazole (kbt) TMPRSS2 inhibitors which are structurally distinct from and have significantly improved activity over the existing known inhibitors Camostat and Nafamostat. Lead compound MM3122 (4) has an IC50 (half-maximal inhibitory concentration) of 340 pM against recombinant full-length TMPRSS2 protein, an EC50 (half-maximal effective concentration) of 430 pM in blocking host cell entry into Calu-3 human lung epithelial cells of a newly developed VSV-SARS-CoV-2 chimeric virus, and an EC50 of 74 nM in inhibiting cytopathic effects induced by SARS-CoV-2 virus in Calu-3 cells. Further, MM3122 blocks Middle East respiratory syndrome coronavirus (MERS-CoV) cell entry with an EC50 of 870 pM. MM3122 has excellent metabolic stability, safety, and pharmacokinetics in mice, with a half-life of 8.6 h in plasma and 7.5 h in lung tissue, making it suitable for in vivo efficacy evaluation and a promising drug candidate for COVID-19 treatment.
KW - Antiviral
KW - COVID-19
KW - PS-SCL
KW - Protease inhibitor
KW - Structure-based drug discovery
UR - http://www.scopus.com/inward/record.url?scp=85117390094&partnerID=8YFLogxK
U2 - 10.1073/pnas.2108728118
DO - 10.1073/pnas.2108728118
M3 - Article
C2 - 34635581
AN - SCOPUS:85117390094
VL - 118
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 43
M1 - e2108728118
ER -