Discovery of three small-molecule inhibitors targeting Ebolavirus genome replication and transcription

The 2013 Ebola virus (EBOV) outbreak was the largest in history. Despite recent advances in both vaccines and monoclonal antibody therapies, 12 years later, EBOV still poses a substantial threat. Previously, we published a ligand discovery pipeline combining <i>in silico</i> screening of compounds with a robust and rapid EBOV minigenome assay for early-stage inhibitor validation at Biological Safety Level 2. Here, we present the further use of this pipeline to identify three compounds that also inhibit EBOV minigenome transcription and replication. They are efficacious in the nM range, exhibited low cytotoxicity and were specific, with no effect on either a T7 RNA polymerase-driven firefly luciferase or a Bunyamwera virus minigenome. Furthermore, these small-molecule inhibitors exhibited the ability to block EBOV minigenome activity when applied after establishment of replication complexes, with implications for potential post-exposure EBOV treatment.