The Anticoagulant Nafamostat Potently Inhibits SARS-CoV-2 S Protein-Mediated Fusion in a Cell Fusion Assay System and Viral Infection In Vitro in a Cell-Type-Dependent Manner
Although infection by SARS-CoV-2, the causative agent of coronavirus pneumonia disease (COVID-19), is spreading rapidly worldwide, no drug has been shown to be sufficiently effective for treating COVID-19. We previously found that nafamostat mesylate, an existing drug used for disseminated intravasc...
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MDPI AG
2020-06-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/12/6/629 |
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| author | Mizuki Yamamoto Maki Kiso Yuko Sakai-Tagawa Kiyoko Iwatsuki-Horimoto Masaki Imai Makoto Takeda Noriko Kinoshita Norio Ohmagari Jin Gohda Kentaro Semba Zene Matsuda Yasushi Kawaguchi Yoshihiro Kawaoka Jun-ichiro Inoue |
| author_facet | Mizuki Yamamoto Maki Kiso Yuko Sakai-Tagawa Kiyoko Iwatsuki-Horimoto Masaki Imai Makoto Takeda Noriko Kinoshita Norio Ohmagari Jin Gohda Kentaro Semba Zene Matsuda Yasushi Kawaguchi Yoshihiro Kawaoka Jun-ichiro Inoue |
| author_sort | Mizuki Yamamoto |
| collection | DOAJ |
| description | Although infection by SARS-CoV-2, the causative agent of coronavirus pneumonia disease (COVID-19), is spreading rapidly worldwide, no drug has been shown to be sufficiently effective for treating COVID-19. We previously found that nafamostat mesylate, an existing drug used for disseminated intravascular coagulation (DIC), effectively blocked Middle East respiratory syndrome coronavirus (MERS-CoV) S protein-mediated cell fusion by targeting transmembrane serine protease 2 (TMPRSS2), and inhibited MERS-CoV infection of human lung epithelium-derived Calu-3 cells. Here we established a quantitative fusion assay dependent on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) S protein, angiotensin I converting enzyme 2 (ACE2) and TMPRSS2, and found that nafamostat mesylate potently inhibited the fusion while camostat mesylate was about 10-fold less active. Furthermore, nafamostat mesylate blocked SARS-CoV-2 infection of Calu-3 cells with an effective concentration (EC)<sub>50</sub> around 10 nM, which is below its average blood concentration after intravenous administration through continuous infusion. On the other hand, a significantly higher dose (EC<sub>50</sub> around 30 μM) was required for VeroE6/TMPRSS2 cells, where the TMPRSS2-independent but cathepsin-dependent endosomal infection pathway likely predominates. Together, our study shows that nafamostat mesylate potently inhibits SARS-CoV-2 S protein-mediated fusion in a cell fusion assay system and also inhibits SARS-CoV-2 infection in vitro in a cell-type-dependent manner. These findings, together with accumulated clinical data regarding nafamostat’s safety, make it a likely candidate drug to treat COVID-19. |
| first_indexed | 2024-03-10T19:15:32Z |
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| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-10T19:15:32Z |
| publishDate | 2020-06-01 |
| publisher | MDPI AG |
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| series | Viruses |
| spelling | doaj.art-ef95b75cf9ad4ebd9a09062a7a03ac9e2023-11-20T03:27:38ZengMDPI AGViruses1999-49152020-06-0112662910.3390/v12060629The Anticoagulant Nafamostat Potently Inhibits SARS-CoV-2 S Protein-Mediated Fusion in a Cell Fusion Assay System and Viral Infection In Vitro in a Cell-Type-Dependent MannerMizuki Yamamoto0Maki Kiso1Yuko Sakai-Tagawa2Kiyoko Iwatsuki-Horimoto3Masaki Imai4Makoto Takeda5Noriko Kinoshita6Norio Ohmagari7Jin Gohda8Kentaro Semba9Zene Matsuda10Yasushi Kawaguchi11Yoshihiro Kawaoka12Jun-ichiro Inoue13Research Center for Asian Infectious Diseases, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, JapanDivision of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, JapanDivision of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, JapanDivision of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, JapanDivision of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, JapanDepartment of Virology 3, National Institute of Infectious Diseases, Musashimurayama, Tokyo 208-0011, JapanDisease Control and Prevention Center, National Center for Global Health and Medicine, Tokyo 162-8655, JapanDisease Control and Prevention Center, National Center for Global Health and Medicine, Tokyo 162-8655, JapanResearch Center for Asian Infectious Diseases, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, JapanDepartment of Life Science and Medical Bio-Science, Waseda University, Shinjuku-ku, Tokyo 162-8480, JapanResearch Center for Asian Infectious Diseases, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, JapanResearch Center for Asian Infectious Diseases, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, JapanDivision of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, JapanResearch Center for Asian Infectious Diseases, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, JapanAlthough infection by SARS-CoV-2, the causative agent of coronavirus pneumonia disease (COVID-19), is spreading rapidly worldwide, no drug has been shown to be sufficiently effective for treating COVID-19. We previously found that nafamostat mesylate, an existing drug used for disseminated intravascular coagulation (DIC), effectively blocked Middle East respiratory syndrome coronavirus (MERS-CoV) S protein-mediated cell fusion by targeting transmembrane serine protease 2 (TMPRSS2), and inhibited MERS-CoV infection of human lung epithelium-derived Calu-3 cells. Here we established a quantitative fusion assay dependent on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) S protein, angiotensin I converting enzyme 2 (ACE2) and TMPRSS2, and found that nafamostat mesylate potently inhibited the fusion while camostat mesylate was about 10-fold less active. Furthermore, nafamostat mesylate blocked SARS-CoV-2 infection of Calu-3 cells with an effective concentration (EC)<sub>50</sub> around 10 nM, which is below its average blood concentration after intravenous administration through continuous infusion. On the other hand, a significantly higher dose (EC<sub>50</sub> around 30 μM) was required for VeroE6/TMPRSS2 cells, where the TMPRSS2-independent but cathepsin-dependent endosomal infection pathway likely predominates. Together, our study shows that nafamostat mesylate potently inhibits SARS-CoV-2 S protein-mediated fusion in a cell fusion assay system and also inhibits SARS-CoV-2 infection in vitro in a cell-type-dependent manner. These findings, together with accumulated clinical data regarding nafamostat’s safety, make it a likely candidate drug to treat COVID-19.https://www.mdpi.com/1999-4915/12/6/629SARS-CoV-2TMPRSS2fusion inhibitor |
| spellingShingle | Mizuki Yamamoto Maki Kiso Yuko Sakai-Tagawa Kiyoko Iwatsuki-Horimoto Masaki Imai Makoto Takeda Noriko Kinoshita Norio Ohmagari Jin Gohda Kentaro Semba Zene Matsuda Yasushi Kawaguchi Yoshihiro Kawaoka Jun-ichiro Inoue The Anticoagulant Nafamostat Potently Inhibits SARS-CoV-2 S Protein-Mediated Fusion in a Cell Fusion Assay System and Viral Infection In Vitro in a Cell-Type-Dependent Manner Viruses SARS-CoV-2 TMPRSS2 fusion inhibitor |
| title | The Anticoagulant Nafamostat Potently Inhibits SARS-CoV-2 S Protein-Mediated Fusion in a Cell Fusion Assay System and Viral Infection In Vitro in a Cell-Type-Dependent Manner |
| title_full | The Anticoagulant Nafamostat Potently Inhibits SARS-CoV-2 S Protein-Mediated Fusion in a Cell Fusion Assay System and Viral Infection In Vitro in a Cell-Type-Dependent Manner |
| title_fullStr | The Anticoagulant Nafamostat Potently Inhibits SARS-CoV-2 S Protein-Mediated Fusion in a Cell Fusion Assay System and Viral Infection In Vitro in a Cell-Type-Dependent Manner |
| title_full_unstemmed | The Anticoagulant Nafamostat Potently Inhibits SARS-CoV-2 S Protein-Mediated Fusion in a Cell Fusion Assay System and Viral Infection In Vitro in a Cell-Type-Dependent Manner |
| title_short | The Anticoagulant Nafamostat Potently Inhibits SARS-CoV-2 S Protein-Mediated Fusion in a Cell Fusion Assay System and Viral Infection In Vitro in a Cell-Type-Dependent Manner |
| title_sort | anticoagulant nafamostat potently inhibits sars cov 2 s protein mediated fusion in a cell fusion assay system and viral infection in vitro in a cell type dependent manner |
| topic | SARS-CoV-2 TMPRSS2 fusion inhibitor |
| url | https://www.mdpi.com/1999-4915/12/6/629 |
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