The ReFRAME library as a comprehensive drug repurposing library to identify mammarenavirus inhibitors

Several mammarenaviruses, chiefly Lassa virus (LASV) in Western Africa and Junín virus (JUNV) in the Argentine Pampas, cause severe disease in humans and pose important public health problems in their endemic regions. Moreover, mounting evidence indicates that the worldwide-distributed mammarenavirus lymphocytic choriomeningitis virus (LCMV) is a neglected human pathogen of clinical significance. The lack of licensed mammarenavirus vaccines and partial efficacy of current anti-mammarenavirus therapy limited to an off-label use of the nucleoside analog ribavirin underscore an unmet need for novel therapeutics to combat human pathogenic mammarenavirus infections. This task can be facilitated by the implementation of “drug repurposing” strategies to reduce the time and resources required to advance identified antiviral drug candidates into the clinic. We screened a drug repurposing library of 11,968 compounds (Repurposing, Focused Rescue and Accelerated Medchem [ReFRAME]) and identified several potent inhibitors of LCMV multiplication that had also strong anti-viral activity against LASV and JUNV. Our findings indicate that enzymes of the rate-limiting steps of pyrimidine and purine biosynthesis, the pro-viral MCL1 apoptosis regulator, BCL2 family member protein and the mitochondrial electron transport complex III, play critical roles in the completion of the mammarenavirus life cycle, suggesting they represent potential druggable targets to counter human pathogenic mammarenavirus infections. •Screening of the ReFRAME library identified potent inhibitors of the mammarenaviruses LCMV, LASV and JUNV.•Inhibitors of key enzymes of de novo pyrimidine biosynthesis have antiviral activity against LCMV, LASV, and JUNV.•Inhibitors of pyrimidine biosynthesis have anti-LCMV activity in cells lacking a functional type I interferon system.•Inhibitors of MCL1 and mitochondrial electron transport complex (mETC) III inhibit LCMV, LASV, and JUNV.