{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T03:30:31Z","timestamp":1790652631770,"version":"4.1.0"},"reference-count":46,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2020,10,30]],"date-time":"2020-10-30T00:00:00Z","timestamp":1604016000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Hilfe f\u00fcr krebskranke Kinder Frankfurt e.V.","award":["n\/a"],"award-info":[{"award-number":["n\/a"]}]},{"name":"Frankfurter Stiftung f\u00fcr krebskranke Kinder","award":["n\/a"],"award-info":[{"award-number":["n\/a"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Severe acute respiratory syndrome virus 2 (SARS-CoV-2) is the cause of the current coronavirus disease 19 (COVID-19) pandemic. Protease inhibitors are under consideration as virus entry inhibitors that prevent the cleavage of the coronavirus spike (S) protein by cellular proteases. Herein, we showed that the protease inhibitor aprotinin (but not the protease inhibitor SERPINA1\/alpha-1 antitrypsin) inhibited SARS-CoV-2 replication in therapeutically achievable concentrations. An analysis of proteomics and translatome data indicated that SARS-CoV-2 replication is associated with a downregulation of host cell protease inhibitors. Hence, aprotinin may compensate for downregulated host cell proteases during later virus replication cycles. Aprotinin displayed anti-SARS-CoV-2 activity in different cell types (Caco2, Calu-3, and primary bronchial epithelial cell air\u2013liquid interface cultures) and against four virus isolates. In conclusion, therapeutic aprotinin concentrations exert anti-SARS-CoV-2 activity. An approved aprotinin aerosol may have potential for the early local control of SARS-CoV-2 replication and the prevention of COVID-19 progression to a severe, systemic disease.<\/jats:p>","DOI":"10.3390\/cells9112377","type":"journal-article","created":{"date-parts":[[2020,10,30]],"date-time":"2020-10-30T21:34:47Z","timestamp":1604093687000},"page":"2377","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":85,"title":["Aprotinin Inhibits SARS-CoV-2 Replication"],"prefix":"10.3390","volume":"9","author":[{"given":"Denisa","family":"Bojkova","sequence":"first","affiliation":[{"name":"Institute for Medical Virology, University Hospital, Goethe University, 60596 Frankfurt am Main, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marco","family":"Bechtel","sequence":"additional","affiliation":[{"name":"Institute for Medical Virology, University Hospital, Goethe University, 60596 Frankfurt am Main, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katie-May","family":"McLaughlin","sequence":"additional","affiliation":[{"name":"School of Biosciences, University of Kent, Canterbury CT2 7NJ, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3068-5624","authenticated-orcid":false,"given":"Jake E.","family":"McGreig","sequence":"additional","affiliation":[{"name":"School of Biosciences, University of Kent, Canterbury CT2 7NJ, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2276-8128","authenticated-orcid":false,"given":"Kevin","family":"Klann","sequence":"additional","affiliation":[{"name":"Faculty of Medicine, Institute of Biochemistry II, Goethe University, 60590 Frankfurt am Main, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8025-2846","authenticated-orcid":false,"given":"Carla","family":"Bellinghausen","sequence":"additional","affiliation":[{"name":"Department of Respiratory Medicine and Allergology, University Hospital, Goethe University, 60590 Frankfurt am Main, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5193-7755","authenticated-orcid":false,"given":"Gernot","family":"Rohde","sequence":"additional","affiliation":[{"name":"Department of Respiratory Medicine and Allergology, University Hospital, Goethe University, 60590 Frankfurt am Main, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Danny","family":"Jonigk","sequence":"additional","affiliation":[{"name":"Institute of Pathology, Hannover Medical School (MHH), 30625 Hannover, Germany"},{"name":"Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), The German Center for Lung Research (Deutsches Zentrum f\u00fcr Lungenforschung, DZL), Hannover Medical School (MHH), 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Braubach","sequence":"additional","affiliation":[{"name":"Institute of Pathology, Hannover Medical School (MHH), 30625 Hannover, Germany"},{"name":"Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), The German Center for Lung Research (Deutsches Zentrum f\u00fcr Lungenforschung, DZL), Hannover Medical School (MHH), 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1802-7565","authenticated-orcid":false,"given":"Sandra","family":"Ciesek","sequence":"additional","affiliation":[{"name":"Institute for Medical Virology, University Hospital, Goethe University, 60596 Frankfurt am Main, Germany"},{"name":"German Center for Infection Research, DZIF, External Partner Site, 60596 Frankfurt am Main, Germany"},{"name":"Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), Branch Translational Medicine und Pharmacology, 60596 Frankfurt am Main, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3832-090X","authenticated-orcid":false,"given":"Christian","family":"M\u00fcnch","sequence":"additional","affiliation":[{"name":"Faculty of Medicine, Institute of Biochemistry II, Goethe University, 60590 Frankfurt am Main, Germany"},{"name":"Frankfurt Cancer Institute, Goethe University, 60596 Frankfurt am Main, Germany"},{"name":"Cardio-Pulmonary Institute, Goethe University, 60590 Frankfurt am Main, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5428-6479","authenticated-orcid":false,"given":"Mark N.","family":"Wass","sequence":"additional","affiliation":[{"name":"School of Biosciences, University of Kent, Canterbury CT2 7NJ, 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study","volume":"395","author":"Chen","year":"2020","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Coronaviridae Study Group of the International Committee on Taxonomy of Viruses (2020). The species Severe acute respiratory syndrome-related coronavirus: Classifying 2019-nCoV and naming it SARS-CoV-2. Nat. Microbiol., 5, 536\u2013544.","DOI":"10.1038\/s41564-020-0695-z"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1016\/S1473-3099(20)30120-1","article-title":"An interactive web-based dashboard to track COVID-19 in real time","volume":"20","author":"Dong","year":"2020","journal-title":"Lancet Infect. Dis."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1016\/S0140-6736(20)30251-8","article-title":"Genomic characterisation and epidemiology of 2019 novel coronavirus: Implications for virus origins and receptor binding","volume":"395","author":"Lu","year":"2020","journal-title":"Lancet"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1038\/s41586-020-2008-3","article-title":"A new coronavirus associated with human respiratory disease in China","volume":"579","author":"Wu","year":"2020","journal-title":"Nature"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"270","DOI":"10.1038\/s41586-020-2012-7","article-title":"A pneumonia outbreak associated with a new coronavirus of probable bat origin","volume":"579","author":"Zhou","year":"2020","journal-title":"Nature"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"727","DOI":"10.1056\/NEJMoa2001017","article-title":"China Novel Coronavirus Investigating and Research Team","volume":"382","author":"Zhu","year":"2020","journal-title":"N. Engl. J. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1038\/s41579-018-0118-9","article-title":"Origin and evolution of pathogenic coronaviruses","volume":"17","author":"Cui","year":"2019","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1016\/j.cell.2020.02.052","article-title":"SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor","volume":"181","author":"Hoffmann","year":"2020","journal-title":"Cell"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"7001","DOI":"10.1073\/pnas.2002589117","article-title":"Enhanced isolation of SARS-CoV-2 by TMPRSS2-expressing cells","volume":"117","author":"Matsuyama","year":"2020","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Hoffmann, M., Schroeder, S., Kleine-Weber, H., M\u00fcller, M.A., Drosten, C., and P\u00f6hlmann, S. (2020). Nafamostat mesylate blocks activation of SARS-CoV-2: New treatment option for COVID-19. Antimicrob. Agents Chemother., 64.","DOI":"10.1128\/AAC.00754-20"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Yamamoto, M., Kiso, M., Sakai-Tagawa, Y., Iwatsuki-Horimoto, K., Imai, M., Takeda, M., Kinoshita, N., Ohmagari, N., Gohda, J., and Semba, K. (2020). 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, 12.","DOI":"10.3390\/v12060629"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"198146","DOI":"10.1016\/j.virusres.2020.198146","article-title":"Scaffold morphing of arbidol (umifenovir) in search of multi-targeting therapy halting the interaction of SARS-CoV-2 with ACE2 and other proteases involved in COVID-19","volume":"289","author":"Choudhary","year":"2020","journal-title":"Virus. Res."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.antiviral.2011.07.014","article-title":"Aprotinin and similar protease inhibitors as drugs against influenza","volume":"92","author":"Zhirnov","year":"2011","journal-title":"Antiviral Res."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.biochi.2017.07.016","article-title":"TMPRSS2: A potential target for treatment of influenza virus and coronavirus infections","volume":"142","author":"Shen","year":"2017","journal-title":"Biochimie"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"L51","DOI":"10.1152\/ajplung.2000.278.1.L51","article-title":"Regulation of SLPI and elafin release from bronchial epithelial cells by neutrophil defensins","volume":"278","author":"Kuijpers","year":"2000","journal-title":"Am. J. Physiol. Lung Cell. Mol. Physiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1278","DOI":"10.1056\/NEJMc2001899","article-title":"Evidence of SARS-CoV-2 Infection in Returning Travelers from Wuhan, China","volume":"382","author":"Hoehl","year":"2020","journal-title":"N. Engl. J. Med."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Toptan, T., Hoehl, S., Westhaus, S., Bojkova, D., Berger, A., Rotter, B., Hoffmeier, K., Cinatl, J., Ciesek, S., and Widera, M. (2020). Optimized qRT-PCR Approach for the Detection of Intra- and Extra-Cellular SARS-CoV-2 RNAs. Int. J. Mol. Sci., 21.","DOI":"10.1101\/2020.04.20.052258"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2045","DOI":"10.1016\/S0140-6736(03)13615-X","article-title":"Glycyrrhizin, an active component of liquorice roots, and replication of SARS-associated coronavirus","volume":"361","author":"Cinatl","year":"2003","journal-title":"Lancet"},{"key":"ref_20","first-page":"323","article-title":"High-dose hydrocortisone reduces expression of the pro-inflammatory chemokines CXCL8 and CXCL10 in SARS coronavirus-infected intestinal cells","volume":"15","author":"Cinatl","year":"2005","journal-title":"Int. J. Mol. Med."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/0022-1759(83)90303-4","article-title":"Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays","volume":"65","author":"Mosmann","year":"1983","journal-title":"J. Immunol. Methods."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1707","DOI":"10.3762\/bjnano.10.166","article-title":"Doxorubicin-loaded human serum albumin nanoparticles overcome transporter-mediated drug resistance in drug-adapted cancer cells","volume":"10","author":"Onafuye","year":"2019","journal-title":"Beilstein J. Nanotechnol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1016\/0166-3542(95)00024-G","article-title":"In vitro inhibition of human cytomegalovirus replication in human foreskin fibroblasts and endothelial cells by ascorbic acid 2-phosphate","volume":"27","author":"Cinatl","year":"1995","journal-title":"Antiviral Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"913","DOI":"10.1016\/j.molcel.2019.11.010","article-title":"Functional Translatome Proteomics Reveal Converging and Dose-Dependent Regulation by mTORC1 and eIF2\u03b1","volume":"77","author":"Klann","year":"2020","journal-title":"Mol. Cell."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"D442","DOI":"10.1093\/nar\/gky1106","article-title":"The PRIDE database and related tools and resources in 2019: Improving support for quantification data","volume":"47","author":"Csordas","year":"2019","journal-title":"Nucleic Acids Res."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"100052","DOI":"10.1016\/j.medidd.2020.100052","article-title":"Inhibition of metalloproteinases in therapy for severe lung injury due to COVID-19","volume":"7","author":"Solun","year":"2020","journal-title":"Med. Drug Discov."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1443","DOI":"10.1056\/NEJMra1910234","article-title":"Alpha(1)-Antitrypsin Deficiency","volume":"382","author":"Strnad","year":"2020","journal-title":"N. Engl. J. Med."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"4751","DOI":"10.1021\/cr010170+","article-title":"Serpin structure, mechanism, and function","volume":"102","author":"Gettins","year":"2002","journal-title":"Chem. Rev."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"981","DOI":"10.1093\/jac\/dkm307","article-title":"Minocycline inhibits West Nile virus replication and apoptosis in human neuronal cells","volume":"60","author":"Michaelis","year":"2007","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"881","DOI":"10.1038\/s41423-020-0485-9","article-title":"The ORF3a protein of SARS-CoV-2 induces apoptosis in cells","volume":"17","author":"Ren","year":"2020","journal-title":"Cell. Mol. Immunol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1101","DOI":"10.1038\/nm.4184","article-title":"Identification of small-molecule inhibitors of Zika virus infection and induced neural cell death via a drug repurposing screen","volume":"22","author":"Xu","year":"2016","journal-title":"Nat. Med."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1038\/s41392-020-00334-0","article-title":"SARS-CoV-2 triggers inflammatory responses and cell death through caspase-8 activation","volume":"5","author":"Li","year":"2020","journal-title":"Signal. Transduct. Target. Ther."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"437","DOI":"10.1002\/hep.27426","article-title":"Transmembrane serine protease TMPRSS2 activates hepatitis C virus infection","volume":"61","author":"Esumi","year":"2015","journal-title":"Hepatology"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/j.virol.2020.01.004","article-title":"SPINT2 inhibits proteases involved in activation of both influenza viruses and metapneumoviruses","volume":"543","author":"Straus","year":"2020","journal-title":"Virology"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"469","DOI":"10.1038\/s41586-020-2332-7","article-title":"Proteomics of SARS-CoV-2-infected host cells reveals therapy targets","volume":"583","author":"Bojkova","year":"2020","journal-title":"Nature"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Azouz, N.P., Klingler, A.M., and Rothenberg, M.E. (2020). Alpha 1 Antitrypsin is an Inhibitor of the SARS-CoV2\u2013Priming Protease TMPRSS2. bioRxiv, preprint.","DOI":"10.1101\/2020.05.04.077826"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"e202000786","DOI":"10.26508\/lsa.202000786","article-title":"TMPRSS2 and furin are both essential for proteolytic activation of SARS-CoV-2 in human airway cells","volume":"3","author":"Bestle","year":"2020","journal-title":"Life Sci. Alliance"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"108254","DOI":"10.1016\/j.celrep.2020.108254","article-title":"Furin Inhibitors Block SARS-CoV-2 Spike Protein Cleavage to Suppress Virus Production and Cytopathic Effects","volume":"33","author":"Cheng","year":"2020","journal-title":"Cell Rep."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1013","DOI":"10.1097\/00000542-199405000-00010","article-title":"Pharmacokinetics of aprotinin in preoperative cardiac surgical patients","volume":"80","author":"Levy","year":"1994","journal-title":"Anesthesiology"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"S50","DOI":"10.1097\/00005344-199600001-00011","article-title":"Reducing thrombin formation during cardiopulmonary bypass: Is there a benefit of the additional anticoagulant action of aprotinin?","volume":"27","author":"Dietrich","year":"1996","journal-title":"J. Cardiovasc. Pharmacol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"506","DOI":"10.1016\/0003-4975(96)00317-7","article-title":"Efficacy of aprotinin with various anticoagulant agents in cardiopulmonary bypass","volume":"62","author":"Terrell","year":"1996","journal-title":"Ann. Thorac. Surg."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1272","DOI":"10.1111\/j.1399-6576.2005.00809.x","article-title":"The effects of aprotinin and tranexamic acid on thrombin generation and fibrinolytic response after cardiac surgery","volume":"49","author":"Kuitunen","year":"2005","journal-title":"Acta Anaesthesiol. Scand."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"2113","DOI":"10.1111\/j.1538-7836.2007.02717.x","article-title":"Evaluation of aprotinin and tranexamic acid in different in vitro and in vivo models of fibrinolysis, coagulation and thrombus formation","volume":"5","author":"Sperzel","year":"2007","journal-title":"J. Thromb. Haemost."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Marchandot, B., Sattler, L., Jesel, L., Matsushita, K., Schini-Kerth, V., Grunebaum, L., and Morel, O. (2020). COVID-19 Related Coagulopathy: A Distinct Entity?. J. Clin. Med., 9.","DOI":"10.3390\/jcm9061651"},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Lega, S., Naviglio, S., Volpi, S., and Tommasini, A. (2020). Recent Insight into SARS-CoV2 Immunopathology and Rationale for Potential Treatment and Preventive Strategies in COVID-19. Vaccines, 8.","DOI":"10.3390\/vaccines8020224"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"e202012642","DOI":"10.15252\/emmm.202012642","article-title":"Rationale for targeting Complement in COVID-19","volume":"12","author":"Polycarpou","year":"2020","journal-title":"EMBO Mol. Med."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/9\/11\/2377\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:27:17Z","timestamp":1760178437000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/9\/11\/2377"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,10,30]]},"references-count":46,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2020,11]]}},"alternative-id":["cells9112377"],"URL":"https:\/\/doi.org\/10.3390\/cells9112377","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,10,30]]}}}