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Rhinoviruses (RVs) and coronaviruses (CoVs) upregulate host cell metabolic pathways including glycolysis to meet their bioenergetic demands for rapid multiplication. Using the glycolysis inhibitor 2-deoxy-D-glucose (2-DG), we confirm the dose-dependent inhibition of minor- and major-receptor group RV replication. We demonstrate that 2-DG suppresses viral positive- as well as negative-strand RNA synthesis, resulting in lower amounts of progeny virus and RV-mediated cell death. In tissue culture with physiologic glucose levels, 2-DG has a pronounced antiviral effect. Further, assessment of 2-DG's intracellular kinetics revealed that the active intermediate, 2-DG6P, is stored intracellularly for several hours. Our concurrent study of 2-DG's impact on pandemic SARS-CoV-2 and endemic HCoVs demonstrated a significant reduction in viral load. Collectively, these results suggest 2-DG to be a broad-spectrum antiviral.

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