Anti-Inflammatory Activities of Ocotillol Isolated from Tetragonula iridipennis Propolis: A Study on In Vitro and In Silico Models

<b>Background/Objectives</b>: This study evaluated the anti-inflammatory effects of ocotillol, a compound isolated from the ethanol extract of propolis of the <i>Tetragonula iridipennis</i> 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. <b>Methods</b>: 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. <b>Results</b>: The in vitro results demonstrated that ocotillol exhibited significant anti-inflammatory effects by effectively inhibiting NO production, with an IC₅₀ 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. <b>Conclusions</b>: 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.