{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T06:41:47Z","timestamp":1791268907447,"version":"4.1.0"},"update-to":[{"DOI":"10.1371\/journal.ppat.1011328","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2023,8,17]],"date-time":"2023-08-17T00:00:00Z","timestamp":1692230400000}}],"reference-count":51,"publisher":"Public Library of Science (PLoS)","issue":"8","license":[{"start":{"date-parts":[[2023,8,7]],"date-time":"2023-08-07T00:00:00Z","timestamp":1691366400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Biotron Limited"},{"DOI":"10.13039\/100019180","name":"HORIZON EUROPE European Research Council","doi-asserted-by":"publisher","award":["682549"],"award-info":[{"award-number":["682549"]}],"id":[{"id":"10.13039\/100019180","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003554","name":"Lundbeck Foundation","doi-asserted-by":"publisher","award":["lR242-2017-409"],"award-info":[{"award-number":["lR242-2017-409"]}],"id":[{"id":"10.13039\/501100003554","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100009708","name":"Novo Nordisk Fonden","doi-asserted-by":"publisher","award":["NNF20OC0062899"],"award-info":[{"award-number":["NNF20OC0062899"]}],"id":[{"id":"10.13039\/501100009708","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plospathogens.org"],"crossmark-restriction":false},"short-container-title":["PLoS Pathog"],"abstract":"<jats:p>\n                    The Coronavirus envelope (E) protein is a small structural protein with ion channel activity that plays an important role in virus assembly, budding, immunopathogenesis and disease severity. The viroporin E is also located in Golgi and ER membranes of infected cells and is associated with inflammasome activation and immune dysregulation. Here we evaluated in vitro antiviral activity, mechanism of action and in vivo efficacy of BIT225 for the treatment of SARS-CoV-2 infection. BIT225 showed broad-spectrum direct-acting antiviral activity against SARS-CoV-2 in Calu3 and Vero cells with similar potency across 6 different virus strains. BIT225 inhibited ion channel activity of E protein but did not inhibit endogenous currents or calcium-induced ion channel activity of TMEM16A in Xenopus oocytes. BIT225 administered by oral gavage for 12 days starting 12 hours before infection completely prevented body weight loss and mortality in SARS-CoV-2 infected K18 mice (100% survival, n = 12), while all vehicle-dosed animals reached a mortality endpoint by Day 9 across two studies (n = 12). When treatment started at 24 hours after infection, body weight loss, and mortality were also prevented (100% survival, n = 5), while 4 of 5 mice maintained and increased body weight and survived when treatment started 48 hours after infection. Treatment efficacy was dependent on BIT225 dose and was associated with significant reductions in lung viral load (3.5 log\n                    <jats:sub>10<\/jats:sub>\n                    ), virus titer (4000 pfu\/ml) and lung and serum cytokine levels. These results validate viroporin E as a viable antiviral target and support the clinical study of BIT225 for treatment and prophylaxis of SARS-CoV-2 infection.\n                  <\/jats:p>","DOI":"10.1371\/journal.ppat.1011328","type":"journal-article","created":{"date-parts":[[2023,8,7]],"date-time":"2023-08-07T13:31:07Z","timestamp":1691415067000},"page":"e1011328","update-policy":"https:\/\/doi.org\/10.1371\/journal.ppat.corrections_policy","source":"Crossref","is-referenced-by-count":13,"title":["Post-infection treatment with the E protein inhibitor BIT225 reduces disease severity and increases survival of K18-hACE2 transgenic mice infected with a lethal dose of SARS-CoV-2"],"prefix":"10.1371","volume":"19","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5919-8275","authenticated-orcid":true,"given":"Gary","family":"Ewart","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"Bobardt","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bo Hjorth","family":"Bentzen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yannan","family":"Yan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Audrey","family":"Thomson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Klaus","family":"Klumpp","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephen","family":"Becker","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mette M.","family":"Rosenkilde","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michelle","family":"Miller","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Philippe","family":"Gallay","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2023,8,7]]},"reference":[{"key":"ppat.1011328.ref001","unstructured":"World_Health_Organization.: Timeline\u2014COVID-19: Available from: https:\/\/www.who.int\/news\/item\/29-06-2020-covidtimeline."},{"key":"ppat.1011328.ref002","unstructured":"COVID-19_Dashboard. by the Center for Sytems Science and Engineering (CSSE) at Johns Hopkins University (JHU). Available from: https:\/\/coronavirus.jhu.edu\/map.html."},{"key":"ppat.1011328.ref003","unstructured":"Worldometers.info. Available from: https:\/\/www.worldometers.info\/coronavirus."},{"issue":"3","key":"ppat.1011328.ref004","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1038\/s41579-020-00459-7","article-title":"Characteristics of SARS-CoV-2 and COVID-19","volume":"19","author":"B Hu","year":"2021","journal-title":"Nature reviews Microbiology"},{"key":"ppat.1011328.ref005","doi-asserted-by":"crossref","first-page":"m3862","DOI":"10.1136\/bmj.m3862","article-title":"Virology, transmission, and pathogenesis of SARS-CoV-2.","volume":"371","author":"M Cevik","year":"2020","journal-title":"BMJ."},{"issue":"2","key":"ppat.1011328.ref006","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1016\/j.ijmmb.2022.02.005","article-title":"Basic virology of SARS-CoV 2","volume":"40","author":"V Ravi","year":"2022","journal-title":"Indian J Med Microbiol"},{"issue":"2","key":"ppat.1011328.ref007","doi-asserted-by":"crossref","first-page":"e0002621","DOI":"10.1128\/mmbr.00026-21","article-title":"Molecular Virology of SARS-CoV-2 and Related Coronaviruses","volume":"86","author":"YA Kung","year":"2022","journal-title":"Microbiol Mol Biol Rev"},{"issue":"2","key":"ppat.1011328.ref008","doi-asserted-by":"crossref","first-page":"835","DOI":"10.1128\/AAC.01308-09","article-title":"Antiviral efficacy of the novel compound BIT225 against HIV-1 release from human macrophages","volume":"54","author":"G Khoury","year":"2010","journal-title":"Antimicrobial agents and chemotherapy"},{"issue":"2","key":"ppat.1011328.ref009","doi-asserted-by":"crossref","first-page":"144","DOI":"10.1016\/j.antiviral.2010.02.312","article-title":"A novel Hepatitis C virus p7 ion channel inhibitor, BIT225, inhibits bovine viral diarrhea virus in vitro and shows synergism with recombinant interferon-alpha-2b and nucleoside analogues","volume":"86","author":"CA Luscombe","year":"2010","journal-title":"Antiviral research"},{"issue":"1","key":"ppat.1011328.ref010","doi-asserted-by":"crossref","first-page":"eaap8258","DOI":"10.1126\/sciadv.aap8258","article-title":"Organelle luminal dependence of (+)strand RNA virus replication reveals a hidden druggable target.","volume":"4","author":"M Nishikiori","year":"2018","journal-title":"Sci Adv"},{"issue":"10","key":"ppat.1011328.ref011","doi-asserted-by":"crossref","first-page":"557","DOI":"10.1089\/dna.2013.2159","article-title":"Viroporins customize host cells for efficient viral propagation","volume":"32","author":"KM Giorda","year":"2013","journal-title":"DNA Cell Biol"},{"issue":"7","key":"ppat.1011328.ref012","doi-asserted-by":"crossref","first-page":"3552","DOI":"10.3390\/v7072786","article-title":"Relevance of Viroporin Ion Channel Activity on Viral Replication and Pathogenesis.","volume":"7","author":"JL Nieto-Torres","year":"2015","journal-title":"Viruses"},{"issue":"8","key":"ppat.1011328.ref013","doi-asserted-by":"crossref","first-page":"563","DOI":"10.1038\/nrmicro2820","article-title":"Viroporins: structure and biological functions.","volume":"10","author":"JL Nieva","year":"2012","journal-title":"Nature reviews Microbiology"},{"key":"ppat.1011328.ref014","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1007\/978-981-10-8456-0_15","article-title":"Beyond Channel Activity: Protein-Protein Interactions Involving Viroporins.","volume":"88","author":"J To","year":"2018","journal-title":"Subcell Biochem."},{"issue":"11","key":"ppat.1011328.ref015","doi-asserted-by":"crossref","DOI":"10.3390\/v13112165","article-title":"Unravelling the Immunomodulatory Effects of Viral Ion Channels, towards the Treatment of Disease.","volume":"13","author":"S Gargan","year":"2021","journal-title":"Viruses."},{"issue":"1","key":"ppat.1011328.ref016","first-page":"1","article-title":"Amantadine inhibits known and novel ion channels encoded by SARS-CoV-2 in vitro.","volume":"4","author":"TL Toft-Bertelsen","year":"2021","journal-title":"Commun Biol"},{"issue":"8","key":"ppat.1011328.ref017","doi-asserted-by":"crossref","first-page":"847","DOI":"10.1038\/s41422-021-00519-4","article-title":"SARS-CoV-2 envelope protein causes acute respiratory distress syndrome (ARDS)-like pathological damages and constitutes an antiviral target.","volume":"31","author":"B Xia","year":"2021","journal-title":"Cell Res"},{"issue":"12","key":"ppat.1011328.ref018","doi-asserted-by":"crossref","first-page":"1202","DOI":"10.1038\/s41594-020-00536-8","article-title":"Structure and drug binding of the SARS-CoV-2 envelope protein transmembrane domain in lipid bilayers","volume":"27","author":"VS Mandala","year":"2020","journal-title":"Nat Struct Mol Biol"},{"key":"ppat.1011328.ref019","doi-asserted-by":"crossref","first-page":"50","DOI":"10.3389\/fmicb.2019.00050","article-title":"Severe Acute Respiratory Syndrome Coronavirus Viroporin 3a Activates the NLRP3","volume":"10","author":"IY Chen","year":"2019","journal-title":"Inflammasome. Frontiers in microbiology"},{"key":"ppat.1011328.ref020","doi-asserted-by":"crossref","first-page":"105738","DOI":"10.1016\/j.biocel.2020.105738","article-title":"Viroporins and inflammasomes: A key to understand virus-induced inflammation","volume":"122","author":"NS Farag","year":"2020","journal-title":"The international journal of biochemistry & cell biology"},{"key":"ppat.1011328.ref021","doi-asserted-by":"crossref","first-page":"330","DOI":"10.1016\/j.virol.2015.08.010","article-title":"Severe acute respiratory syndrome coronavirus E protein transports calcium ions and activates the NLRP3 inflammasome","volume":"485","author":"JL Nieto-Torres","year":"2015","journal-title":"Virology"},{"key":"ppat.1011328.ref022","doi-asserted-by":"crossref","first-page":"1021","DOI":"10.3389\/fimmu.2020.01021","article-title":"Novel Coronavirus-Induced NLRP3 Inflammasome Activation: A Potential Drug Target in the Treatment of COVID-19","volume":"11","author":"A. Shah","year":"2020","journal-title":"Front Immunol."},{"issue":"5","key":"ppat.1011328.ref023","doi-asserted-by":"crossref","first-page":"e1004077","DOI":"10.1371\/journal.ppat.1004077","article-title":"Severe acute respiratory syndrome coronavirus envelope protein ion channel activity promotes virus fitness and pathogenesis.","volume":"10","author":"JL Nieto-Torres","year":"2014","journal-title":"PLoS Pathog"},{"issue":"3","key":"ppat.1011328.ref024","doi-asserted-by":"crossref","DOI":"10.1128\/mBio.02325-17","article-title":"Role of Severe Acute Respiratory Syndrome Coronavirus Viroporins E, 3a, and 8a in Replication and Pathogenesis.","volume":"9","author":"C Castano-Rodriguez","year":"2018","journal-title":"mBio."},{"issue":"1","key":"ppat.1011328.ref025","doi-asserted-by":"crossref","first-page":"322","DOI":"10.1016\/j.virol.2004.09.033","article-title":"SARS coronavirus E protein forms cation-selective ion channels","volume":"330","author":"L Wilson","year":"2004","journal-title":"Virology"},{"key":"ppat.1011328.ref026","doi-asserted-by":"crossref","first-page":"573","DOI":"10.1007\/978-0-387-33012-9_104","article-title":"Validation of coronavirus E proteins ion channels as targets for antiviral drugs","volume":"581","author":"L Wilson","year":"2006","journal-title":"Adv Exp Med Biol"},{"issue":"2","key":"ppat.1011328.ref027","doi-asserted-by":"crossref","first-page":"294","DOI":"10.1016\/j.virol.2006.05.028","article-title":"Hexamethylene amiloride blocks E protein ion channels and inhibits coronavirus replication","volume":"353","author":"L Wilson","year":"2006","journal-title":"Virology"},{"issue":"7830","key":"ppat.1011328.ref028","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1038\/s41586-020-2787-6","article-title":"Animal models for COVID-19","volume":"586","author":"C Munoz-Fontela","year":"2020","journal-title":"Nature"},{"key":"ppat.1011328.ref029","doi-asserted-by":"crossref","first-page":"146387","DOI":"10.1016\/j.gene.2022.146387","article-title":"Role of genomics in combating COVID-19 pandemic","volume":"823","author":"KA Saravanan","year":"2022","journal-title":"Gene"},{"issue":"1","key":"ppat.1011328.ref030","doi-asserted-by":"crossref","first-page":"177","DOI":"10.3233\/TUB-200084","article-title":"The expression of hACE2 receptor protein and its involvement in SARS-CoV-2 entry, pathogenesis, and its application as potential therapeutic target","volume":"43","author":"L Al-Zaidan","year":"2021","journal-title":"Tumour Biol"},{"issue":"15","key":"ppat.1011328.ref031","doi-asserted-by":"crossref","first-page":"7264","DOI":"10.1128\/JVI.00737-08","article-title":"Severe acute respiratory syndrome coronavirus infection causes neuronal death in the absence of encephalitis in mice transgenic for human ACE2","volume":"82","author":"J Netland","year":"2008","journal-title":"J Virol"},{"issue":"2","key":"ppat.1011328.ref032","doi-asserted-by":"crossref","first-page":"813","DOI":"10.1128\/JVI.02012-06","article-title":"Lethal infection of K18-hACE2 mice infected with severe acute respiratory syndrome coronavirus","volume":"81","author":"PB McCray","year":"2007","journal-title":"J Virol"},{"issue":"11","key":"ppat.1011328.ref033","doi-asserted-by":"crossref","first-page":"1327","DOI":"10.1038\/s41590-020-0778-2","article-title":"SARS-CoV-2 infection of human ACE2-transgenic mice causes severe lung inflammation and impaired function","volume":"21","author":"ES Winkler","year":"2020","journal-title":"Nat Immunol"},{"issue":"7843","key":"ppat.1011328.ref034","doi-asserted-by":"crossref","first-page":"603","DOI":"10.1038\/s41586-020-2943-z","article-title":"COVID-19 treatments and pathogenesis including anosmia in K18-hACE2 mice","volume":"589","author":"J Zheng","year":"2021","journal-title":"Nature"},{"issue":"1","key":"ppat.1011328.ref035","doi-asserted-by":"crossref","first-page":"6122","DOI":"10.1038\/s41467-020-19891-7","article-title":"Lethality of SARS-CoV-2 infection in K18 human angiotensin-converting enzyme 2 transgenic mice.","volume":"11","author":"FS Oladunni","year":"2020","journal-title":"Nat Commun."},{"issue":"19","key":"ppat.1011328.ref036","doi-asserted-by":"crossref","DOI":"10.1172\/jci.insight.142032","article-title":"Human angiotensin-converting enzyme 2 transgenic mice infected with SARS-CoV-2 develop severe and fatal respiratory disease.","volume":"5","author":"JW Golden","year":"2020","journal-title":"JCI Insight."},{"key":"ppat.1011328.ref037","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1186\/s12981-016-0093-z","article-title":"The antiviral compound BIT225 inhibits HIV-1 replication in myeloid dendritic cells","volume":"13","author":"G Khoury","year":"2016","journal-title":"AIDS Res Ther"},{"issue":"7","key":"ppat.1011328.ref038","doi-asserted-by":"crossref","first-page":"2559","DOI":"10.1007\/s00216-010-3498-x","article-title":"Ligand-protein docking studies of potential HIV-1 drug compounds using the algorithm FlexX","volume":"396","author":"G Patargias","year":"2010","journal-title":"Analytical and bioanalytical chemistry"},{"issue":"3","key":"ppat.1011328.ref039","doi-asserted-by":"crossref","first-page":"731","DOI":"10.1093\/jac\/dkv389","article-title":"A Phase 1b\/2a study of the safety, pharmacokinetics and antiviral activity of BIT225 in patients with HIV-1 infection","volume":"71","author":"J Wilkinson","year":"2016","journal-title":"J Antimicrob Chemother"},{"issue":"11","key":"ppat.1011328.ref040","doi-asserted-by":"crossref","first-page":"1914","DOI":"10.1093\/infdis\/jiaa635","article-title":"Human Immunodeficiency Virus Type 1 Vpu Inhibitor, BIT225, in Combination with 3-Drug Antiretroviral Therapy: Inflammation and Immune Cell Modulation","volume":"223","author":"CA Luscombe","year":"2021","journal-title":"The Journal of infectious diseases"},{"issue":"396","key":"ppat.1011328.ref041","doi-asserted-by":"crossref","DOI":"10.1126\/scitranslmed.aal3653","article-title":"Broad-spectrum antiviral GS-5734 inhibits both epidemic and zoonotic coronaviruses","volume":"9","author":"TP Sheahan","year":"2017","journal-title":"Science translational medicine"},{"issue":"541","key":"ppat.1011328.ref042","doi-asserted-by":"crossref","DOI":"10.1126\/scitranslmed.abb5883","article-title":"An orally bioavailable broad-spectrum antiviral inhibits SARS-CoV-2 in human airway epithelial cell cultures and multiple coronaviruses in mice","volume":"12","author":"TP Sheahan","year":"2020","journal-title":"Science translational medicine"},{"issue":"7850","key":"ppat.1011328.ref043","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1038\/s41586-021-03312-w","article-title":"SARS-CoV-2 infection is effectively treated and prevented by EIDD-2801","volume":"591","author":"A Wahl","year":"2021","journal-title":"Nature"},{"key":"ppat.1011328.ref044","doi-asserted-by":"crossref","first-page":"105430","DOI":"10.1016\/j.antiviral.2022.105430","article-title":"Combination therapy with nirmatrelvir and molnupiravir improves the survival of SARS-CoV-2 infected mice","volume":"208","author":"JH Jeong","year":"2022","journal-title":"Antiviral research"},{"issue":"29","key":"ppat.1011328.ref045","doi-asserted-by":"crossref","DOI":"10.1073\/pnas.2101555118","article-title":"Postinfection treatment with a protease inhibitor increases survival of mice with a fatal SARS-CoV-2 infection","volume":"118","author":"CS Dampalla","year":"2021","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"1","key":"ppat.1011328.ref046","doi-asserted-by":"crossref","first-page":"24432","DOI":"10.1038\/s41598-021-04133-7","article-title":"Modulation of the NLRP3 inflammasome by Sars-CoV-2 Envelope protein.","volume":"11","author":"M Yalcinkaya","year":"2021","journal-title":"Scientific reports."},{"issue":"7908","key":"ppat.1011328.ref047","doi-asserted-by":"crossref","first-page":"146","DOI":"10.1038\/s41586-022-04630-3","article-title":"Eicosanoid signalling blockade protects middle-aged mice from severe COVID-19","volume":"605","author":"LR Wong","year":"2022","journal-title":"Nature"},{"issue":"8","key":"ppat.1011328.ref048","article-title":"Protective Effects of Astodrimer Sodium 1% Nasal Spray Formulation against SARS-CoV-2 Nasal Challenge in K18-hACE2 Mice.","volume":"13","author":"JRA Paull","year":"2021","journal-title":"Viruses."},{"key":"ppat.1011328.ref049","doi-asserted-by":"crossref","first-page":"105089","DOI":"10.1016\/j.antiviral.2021.105089","article-title":"Virucidal and antiviral activity of astodrimer sodium against SARS-CoV-2 in vitro","volume":"191","author":"JRA Paull","year":"2021","journal-title":"Antiviral research"},{"key":"ppat.1011328.ref050","volume-title":"R: A language and 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