Identification of broad anti-coronavirus chemical agents for repurposing against SARS-CoV-2 and variants of concern

• Published in: Current Research in Virological Science 2022, Vol.3, p.100019-100019, Article 100019
• Main Authors: Murer, Luca, Volle, Romain, Andriasyan, Vardan, Petkidis, Anthony, Gomez-Gonzalez, Alfonso, Yang, Liliane, Meili, Nicole, Suomalainen, Maarit, Bauer, Michael, Policarpo Sequeira, Daniela, Olszewski, Dominik, Georgi, Fanny, Kuttler, Fabien, Turcatti, Gerardo, Greber, Urs F.
• Format: Article
• Language: English
• Subjects: Drug synergy in vitro; Human coronavirus drug screen at single cell resolution in arrayed format; Human explant nasal and bronchial epithelial cells; Mycophenolic acid; Posaconazole; Post-entry coronavirus inhibition; Repurposing methylene blue; SARS-CoV-2 variants of concern
• ISSN: 2666-478X; 2666-478X
• DOI: 10.1016/j.crviro.2022.100019

Endemic human coronaviruses (hCoVs) 229E and OC43 cause respiratory disease with recurrent infections, while severe acute respiratory syndrome (SARS)-CoV-2 spreads across the world with impact on health and societies. Here, we report an image-based multicycle infection procedure with α-coronavirus hCoV-229E-eGFP in an arrayed chemical library screen of 5440 clinical and preclinical compounds. Toxicity counter selection and challenge with the β-coronaviruses OC43 and SARS-CoV-2 in tissue culture and human airway epithelial explant cultures (HAEEC) identified four FDA-approved compounds with oral availability. Methylene blue (MB, used for the treatment of methemoglobinemia), Mycophenolic acid (MPA, used in organ transplantation) and the anti-fungal agent Posaconazole (POS) had the broadest anti-CoV spectrum. They inhibited the shedding of SARS-CoV-2 and variants-of-concern (alpha, beta, gamma, delta) from HAEEC in either pre- or post exposure regimens at clinically relevant concentrations. Co-treatment of cultured cells with MB and the FDA-approved SARS-CoV-2 RNA-polymerase inhibitor Remdesivir reduced the effective anti-viral concentrations of MB by 2-fold, and Remdesivir by 4 to 10-fold, indicated by BLISS independence synergy modelling. Neither MB, nor MPA, nor POS affected the cell delivery of SARS-CoV-2 or OC43 (+)sense RNA, but blocked subsequent viral RNA accumulation in cells. Unlike Remdesivir, MB, MPA or POS did not reduce the release of viral RNA in post exposure regimen, thus indicating infection inhibition at a post-replicating step as well. In summary, the data emphasize the power of unbiased, full cycle compound screens to identify and repurpose broadly acting drugs against coronaviruses. [Display omitted] •A novel imaging-based multicycle platform screened 5440 chemicals against human coronavirus cell culture infections.•Identification of five FDA-approved small chemical compounds to repurpose against COVID-19.•MB, MPA, POS are orally available, and inhibit infection of SARS-CoV-2 in human airway epithelial explant cultures.•MB synergizes with the RNA-dependent RNA polymerase inhibitor Remdesivir against coronavirus infection in cell culture.•d MB and MPA ​offer ​two-pronged ​strategies ​against COVID-19 ​with ​anti-viral and anti-inflammatory ​arms, and ​drug synergy option