Dual-Target Inhibition of Puumala Virus Entry Glycoproteins Gn and Gc by Moringa oleifera -Derived Phytochemicals: An Integrated Computational Study

Puumala virus (PUUV), a member of the Orthohantavirus genus, causes nephropathia epidemica, an intermediate type of hemorrhagic fever with renal dysfunction, yet no approved antiviral therapy is currently available. Viral entry is initiated by the coordinated actions of the attachment glycoprotein Gn and fusion glycoprotein Gc, making simultaneous inhibition of these proteins an attractive antiviral strategy. Here, we employed a combined computational workflow to identify dual-target inhibitors of PUUV entry glycoproteins from Moringa oleifera phytochemicals. 72 phytochemicals were retrieved and docked by molecular docking to identify high-affinity binders, while he top-ranked compounds were subsequently evaluated for drug-likeness, pharmacokinetic behavior, toxicity, and predicted biological activities to assess their therapeutic suitability. Complex stability and binding mechanisms were further investigated through 100 ns molecular dynamics simulations (MDS) and MM-GBSA binding free-energy calculations. Quercetin and kaempferol showed the lowest binding energies toward Gn (−9.8 and −9.7 kcal/mol) and Gc (−10.4 and −10.0 kcal/mol), respectively. Both phytochemicals demonstrated favorable drug-like properties, high gastrointestinal absorption, no predicted toxicity, and positive bioactivity scores. MDS confirmed stable protein–ligand complexes with minimal structural fluctuations, and MM-GBSA analysis further validated strong binding, with quercetin exhibiting the most favorable binding free energies against Gn (−84.24 kcal/mol) and Gc (−98.45 kcal/mol). These findings identify quercetin and kaempferol as promising dual-target inhibitors that may simultaneously disrupt PUUV attachment and membrane fusion, warranting further experimental validation.