Identification of Repurposed Drugs Against Monkeypox Virus Through Comprehensive Virtual Screening.

The recent spread of human monkeypox (mpox) beyond Africa has attracted global attention. Although mpox is a DNA virus with relatively lower pandemic potential, and smallpox vaccines and therapeutics may offer protection, the distinct clinical features observed in the current outbreak highlight the need for novel antiviral drugs. Here, we developed a drug repurposing workflow based on interspecies protein analysis and advanced virtual screening, integrating pre- and post-docking processes. After screening 6,074 approved or clinical-stage drugs against viral DNA topoisomerase, we expanded the analysis to viral DNA polymerase, increasing the compound library to 11,484 molecules, including approved or clinical-stage drugs and DNA polymerase inhibitors from ChEMBL. To assess dynamic stability, we performed molecular dynamics (MD) simulations and quantified protein-ligand contact occupancy, distinguishing stably bound candidates from transient docking poses. The MD and convolutional neural-network (GNINA) scoring analyses converged on vindesine for DNA topoisomerase and LY2090314 for DNA polymerase. Importantly, the workflow was applied to targets with distinct prior information, including a structurally characterized DNA polymerase supported by homologue inhibitor data and a less-characterized DNA topoisomerase requiring homology modelling. These results illustrate an adaptable repurposing pipeline for prioritizing therapeutic candidates against emerging infectious diseases, although experimental validation remains required.