Unveiling the anti-dengue potential of Cedrus deodar bark extract through the network pharmacology, in silico, in vivo, and in vitro strategies

This study integrates network pharmacology, in silico and biological validation to explore the anti-dengue potential of the ethanol extract of Cedrus deodara bark (EECD). Cedrus deodara bark was authenticated, shade-dried, defatted using petroleum ether and extracted with ethanol using Soxhlet extraction. Network pharmacology was performed on key phytoconstituents, Cedrol, Himachalol, Widdrol, Deodarin, Kaempferol, Gallic acid, β-Sitosterol using Swiss target prediction, IMPPAT database, while disease genes were retrieved from GeneCards. Overlapping targets were visualized via Cytoscape, and enriched pathways were identified through KEGG and GO analysis. For in silico validation, ADMET profiling was done via SwissADME and Qikprop to access drug-likeness, oral bioavailability, and toxicity. Compounds with favorable properties were docked using Glide module of Schrodinger against both viral (NS5 RdRp, NS3 protease) and host (TNF-α, COX-2 receptor) targets. The ligand-protein complexes were further refined with Prime MM-GBSA to calculate binding free energy (ΔG_bind). Molecular dynamics (MD) simulation using Desmond for 100 ns were conducted to access RMSD, RMSF, protein ligand interactions and structural stability, In vitro anti-dengue activity was evaluated using the MTT assay on DENV-2-infected Vero cells and percentage growth inhibition were determined. Subsequently, in vivo, hydroxyurea-induced thrombocytopenia model in albino rat was used to access hematological parameters including platelet count, RBC, WBC, hemoglobin, PCV, MCV, MCH and MCHC were evaluated along with bone marrow histopathology was also performed post-treatments. Network pharmacology and pathway analysis revealed the multi-target interactions linked to viral inhibition and immune modulation. Molecular docking results showed binding energies between -9.962 to -10.161 kcal/mol. MD simulations confirmed the stability of key ligand target complexes over 100ns. EECD showed significant cytoprotection in in vitro and normalized hematological balance in thrombocytopenic rats in in vivo. Cedrus deodara ethanol extract demonstrates promising anti-dengue activity, further drug development as a plant-based therapeutic candidate. This is the first comprehensive exploration of Cedrus deodara bark against dengue, combining molecular docking, dynamics, and experimental validation.