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

• Published in: PLoS pathogens Vol. 19; no. 8; p. e1011328
• Main Authors: Ewart, Gary, Bobardt, Michael, Bentzen, Bo Hjorth, Yan, Yannan, Thomson, Audrey, Klumpp, Klaus, Becker, Stephen, Rosenkilde, Mette M, Miller, Michelle, Gallay, Philippe
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 07-08-2023
• Subjects: Antiviral activity; Antiviral agents; Biology and life sciences; Body weight; Body weight loss; Calcium currents; Care and treatment; Cell culture; Channel gating; Coronaviruses; COVID-19; Drug dosages; Drug resistance; Drug therapy; Effectiveness; Env protein; Evaluation; Gametocytes; Genetic engineering; Golgi apparatus; Health aspects; Health services; Immunopathogenesis; Infection; Infections; Inflammasomes; Inflammation; Ion channels; Ions; Lethal dose; Lungs; Medicine and health sciences; Mortality; Oocytes; Physical Sciences; Prevention; Prophylaxis; Proteins; Public health; Public health administration; Research and Analysis Methods; Severe acute respiratory syndrome coronavirus 2; Survival; Transgenic animals; Transgenic mice; Vero cells; Viral diseases; Viral infections; Viruses; Weight loss; Germany
• ISSN: 1553-7374; 1553-7366; 1553-7374
• DOI: 10.1371/journal.ppat.1011328

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 log10), 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.