Dengue　virus　(DENV)　is　a　significant　significant　mosquito-borne　pathogen.　Its　RNA-dependent　RNA　polymerase　(RdRp)　plays　a　crucial　role　in　viral　replication,　making　it　a　critical　target　for　antiviral　drug　development.　The　goal　of　this　study　was　to　identify　potential　inhibitors　of　the　DENV　RdRp　through　virtual　screening　and　experimental　assays.　In　this　study,　based　on　Topscience　and　TargetMol　database.　47　compounds　were　stand　out　from　1.1　million　compounds　with　high　binding　energies　between　−77.24　kcal/mol　and−44.08　kcal/mol　by　initial　virtual　screening.　Their　inhibition　activity　against　dengue　virus　was　evaluated　by　cell-based　antiviral　and　RdRp　enzyme　assays,　and　4　compounds　(R2,　R29,　R37,　and　R39)　identified　in　our　study　demonstrated　promising　in　vitro　activity,　with　IC₅₀　values　of　less　than　10　μM,　indicating　their　strong　potential　as　antiviral　agents　against　DENV.　The　RdRp　enzyme　inhibition　assay　revealed　that　R37　demonstrated　significant　activity　against　the　DENV　RdRp　enzyme,　with　an　IC₅₀　of　10.86　±　2.06　μM,　which　was　lower　than　that　of　3’-dATP　(IC₅₀　=　30.09　±　8.26　μM).　These　4　compounds　were　further　subjected　to　DFT　calculations　and　MD　simulation,　which　not　only　provided　valuable　structural　insights　but　also　enhanced　the　reliability　of　our　findings　by　offering　a　detailed　understanding　of　the　stability　and　binding　interactions　of　the　top　compounds　with　the　DENV　RdRp　enzyme.　These　results　suggest　that　the　identified　compounds　are　promising　candidates　for　novel　DENV　RdRp　inhibitors　and　provide　both　theoretical　and　experimental　groundwork　for　further　optimization　and　validation.　©　2025　Elsevier　B.V.