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4'-fluorouridine is a potent inhibitor of Oropouche virus in vitro and in animal infection models

Ferrie, M.; Darmuzey, M.; Tarillon, I.; Tubiana, T.; Khan, M.; Roskams, T.; Weynand, B.; Thal, D.; Cremers, N.; Hendrickx, S.; Donckers, K.; Portal, T. M.; Vanmechelen, B.; Lemmens, V.; Rocha-Pereira, J.; Castilletti, C.; Mombaerts, P.; Bressanelli, S.; Laporte, M.; MALET, H.; Neyts, J.

2026-09-01 microbiology
10.1101/2025.09.22.677733 bioRxiv
Show abstract

Oropouche virus (OROV) is an orthobunyavirus that causes increasingly frequent and severe outbreaks in Central and South America. We report that 4'-fluorouridine (4'-FlU) inhibits the in vitro replication of epidemic and pre-epidemic OROV strains in multiple cell lines. In vitro polymerase assays demonstrate that 4'-FlU (as its triphosphate) targets the Peribunyaviridae L protein, is incorporated during RNA synthesis and causes premature chain termination. Following 69 consecutive days of in vitro passages of OROV in the presence of suboptimal concentrations of 4'-FlU, no drug-resistant variants were identified in the viral polymerase. In stringent mouse (AG129) or Syrian hamster OROV-infection models, oral administration of 4'-FlU completely blocked viral replication and virus-induced disease, even when administration was delayed until 72 hours after infection. Our findings support exploring the potential of 4'-FlU for the management of OROV infections in humans.

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