Fatigue is a widespread condition associated with various health issues, yet identifying specific bioactive compounds for its management remains challenging. This study integrates network pharmacology and molecular docking to uncover essential oil-derived compounds with potential antifatigue properties by targeting key genes and molecular pathways. A comprehensive analysis of 872 essential oil compounds was conducted using PubChem, with target prediction via SwissTargetPrediction. The protein-protein interaction (PPI) network and KEGG pathway analysis identified core fatigue-related targets, including ALB, BCL2, EGFR, IL-6, and STAT3, in metabolic dysregulation and inflammatory responses linked to fatigue. Molecular docking exhibits strong binding affinity between key compounds such as Calamenene, T-cadinol, and Bornyl acetate and core targets, suggesting their potential antifatigue effects. However, ADMET analysis confirmed T-cadinol’s drug-likeness, suggesting good bioavailability and minimal toxicity risks. Thus, molecular docking revealed high binding affinity, which was further validated through a 100 ns MD simulation and demonstrated stable interactions with low root mean square deviation (RMSD). Additionally, hydrogen bond analysis confirmed that T-cadinol maintained consistent interactions with key residues such as Thr-790 in EGFR, Arg-222 in ALB, and Arg-104 in IL-6, indicating strong binding stability. While this study provides valuable computational insights, further in vitro and in vivo validation is necessary to confirm these findings and explore potential therapeutic applications.

The prostaglandin E2 receptor EP3 is emerging as a promising therapeutic target in cardiovascular diseases because of its involvement in vascular inflammation, platelet aggregation, and vasoconstriction. However, selective EP3 ligands with validated biological activities are scarce. In this study, we combined computational and experimental strategies to identify and validate novel EP3 receptor ligands with therapeutic potential. We implemented a high-throughput, structure- and ligand-based virtual screening pipeline, enabling efficient exploration of approved drugs and natural compounds from DrugBank and FooDB libraries. Top-scoring candidates were prioritised based on binding energy and pharmacophoric similarity. Selected hits were subjected to in silico ADME/Tox profiling using QikProp to identify molecules with favourable pharmacokinetic and safety parameters. TUCA, masoprocol, and pravastatin sodium have emerged as lead candidates and were validated in vitro using endothelial migration and platelet aggregation assays. TUCA exhibited the most consistent inhibitory effect on endothelial migration, whereas masoprocol and hydrocortisone significantly reduced platelet aggregation. These findings establish a multidimensional workflow for the rational identification of EP3 ligands and support their potential use in cardiovascular therapeutics.

The prostaglandin E2 receptor EP3 is emerging as a promising therapeutic target in cardiovascular disease due to its involvement in vascular inflammation, platelet aggregation, and vasoconstriction. However, selective EP3 ligands with validated biological activity remain scarce. Here, we combine computational and experimental strategies to discover and validate novel EP3 receptor ligands with therapeutic potential. We implemented a high-throughput, structure- and ligand- based virtual screening pipeline, enabling the efficient exploration of approved drugs and natural compounds from DrugBank and FooDB libraries. Top-scoring candidates were prioritized based on binding energy, and pharmacophoric similarity. Selected hits underwent in silico ADME/Tox profiling using QikProp, identifying molecules with favorable pharmacokinetic and safety parameters. TUCA, masoprocol, and pravastatin sodium emerged as lead candidates and were validated in vitro using endothelial migration and platelet aggregation assays. TUCA exhibited the most consistent inhibitory effect on endothelial migration, while masoprocol, and hydrocortisone significantly reduced platelet aggregation. These findings establish a multidimensional workflow for the rational identification of EP3 ligands and support their potential in cardiovascular therapeutics.

Odorant-binding proteins (OBPs) play central roles in insect chemoreception and represent attractive molecular targets for the discovery of novel behavior-modifying compounds. We report two crystal structures of Anopheles gambiae OBP4: one complexed with the fluorescent probe N-phenyl-1-naphthylamine and a second containing two indole molecules simultaneously bound within the ligand-binding cavity. Structural comparison revealed two binding subsites accommodating the aromatic moieties of 1-NPN or two indole molecules, while identifying Thr58 and Thr70 as key residues in ligand recognition. A structure-based virtual screening campaign was performed against a volatility-filtered library from DrugBank, FooDB, BioNet, and natural-volatiles datasets. Twenty-three ligands were selected for fluorescence competitive binding assays, identifying four AgamOBP4 binders with micromolar affinities ( K i = 2.4-21.8 μM). Among them were benzoyl peroxide, an active anti-acne ingredient, Avex-73 (blarcamesine), a clinical-stage drug candidate, and 2-phenylethyl benzoate, a food- and fragrance-associated ester. Additional candidates include o,p′-DDE, a metabolite of the insecticide DDT, and fluorene, a polycyclic aromatic hydrocarbon detected in foods as environmental or processing-derived contaminants. Seven selected compounds were subsequently evaluated against Aedes albopictus in in-vivo behavioral assays, with benzoyl peroxide displaying the highest repellency (RI = 81.8% at 0.2 mg cm ^-2 ). AI-based protein-ligand modeling provided insights into ligand recognition and suggested relationships among binding mode, binding affinity, and bioactivity. Collectively, these findings establish the structural basis of ligand recognition by AgamOBP4, highlight the potential of drug-repurposing and food-derived chemical space as sources of next-generation mosquito repellents, and open new opportunities for the development of OBP4-based biosensors for food-quality and environmental monitoring applications. Highlights: Novel AgamOBP4 crystal complexes revealed a dual aromatic-recognition binding site Structure-based virtual screening of >139,000 compounds from DrugBank, FooDB, and BioNet uncovered novel AgamOBP4 ligands from drug-repurposing and food-derived chemical spaces In vitro validation identified the clinical-stage candidate Avex-73 and the anti-acne agent benzoyl peroxide as novel low-micromolar AgamOBP4 ligands. Binding of the DDT metabolite o,p′-DDE suggests a possible involvement of AgamOBP4 in insecticide recognition. Recognition of food-associated fragrances and environmental contaminants highlights the potential for the development of AgamOBP4-based biosensors for food quality and environmental monitoring. Behavioral bioassays have identified benzoyl peroxide as a potent repellent against the Asian tiger mosquito Aedes albopictus . AI-based protein-ligand modeling provided insight into binding-mode, affinity, and bioactivity relationships