Abstract
Background/Objectives: This study evaluated the anti-inflammatory effects of ocotillol, a compound isolated from the ethanol extract of propolis of the Tetragonula iridipennis stingless bee. Through its ability to inhibit NO production in an in vitro model, it investigated the NO inhibition mechanism using network pharmacology combined with molecular docking. Methods: The NO production inhibitory activity was determined by colorimetric assay using Griess reagent. An in silico study was performed using network pharmacology analysis, molecular docking, and molecular dynamics simulations. Results: The in vitro results demonstrated that ocotillol exhibited significant anti-inflammatory effects by effectively inhibiting NO production, with an IC(50) value of 20.29 ± 2.1 µg/mL. The network pharmacology analysis revealed that ocotillol targets 14 molecular sites related to NO, with TACR1 showing the best binding affinity at -10.0 kcal/mol. Molecular dynamics simulations suggest that TACR1 is a potential target. As indicated by the stable interaction profile, further validation in complex biological membranes is warranted. Conclusions: This study also provides evidence for the correlation between in vitro and in silico models, thus laying the groundwork for in vivo evaluations to confirm the anti-inflammatory mechanism of ocotillol.