Exploring the viral protease inhibitor space driven by consensus scoring-based virtual screening.

Dengue fever presents a major health concern, and the lack of an effective vaccine or definite therapeutic regimen has led the research community to identify safe-by-design potential targets for drug discovery. Since the association of the NS2B co-factor with the protease domain of NS3 is imperative for the catalytic activity of the enzyme complex, inhibitors blocking their interaction could provide an alternative strategy to combat the dengue virus. In this context, the present study is aimed at exploring computer-assisted modeling of significant physicochemical features required for the inhibition of the dengue virus protease complex. First of all, alanine scanning was utilized to map hot spot residues critical for the association of the two subunits, NS2B and NS3pro, by studying their energy profiles. Then, consensus score-based virtual screening was performed to search through the commercially available chemical datasets. After screening, 1,575 small molecules were moved forward into docking studies to investigate their interactions with crucial interfacial residues (i.e., Tyr23, Lys26, Phe46, and Leu58), with only 233 molecules passing that stage. The top 30 molecules were selected based on a detailed profile of intermolecular interactions. After that, the top five molecules were selected for detailed mechanistic studies via molecular dynamics simulations followed by subsequent binding free energy calculations, principal component analysis in conjunction with free energy landscape. To the best of our knowledge, this is the first systematic and comprehensive investigation to identify protein-protein interaction blockers against the target protein at such a large scale, using integrated computational tools. Our results highlight the enhanced stability and good binding affinities towards the target protein of these compounds, which might act as new scaffolds for NS2B-NS3 protease inhibition. Future studies will be directed to explore the detailed atomistic-based structural and energetic framework of the mutation-induced affinity change between the protease domain of the DENV-2 NS3 protein and its cofactor NS2B. The detailed insight in turn might suggest precise and focused targeted points for the structure-based drug design but the computational cost may be a challenge. Graphical abstract: Supplementary Information: The online version contains supplementary material available at 10.1007/s40203-023-00174-0.