Inhibitors of Nitric Oxide Production from the Seeds of Myristica Fragrans: Experimental and Computational Results

This study focuses on the extraction of eleven compounds from Myristica fragrans Houtt., Miristicaceae, seeds, guided by their anti-inflammatory properties. Zuonin A, aristolignan, and oleiferin F are reported from M. fragrans for the first time. The anti-inflammatory potential of all isolated compounds was evaluated against lipopolysaccharide (LPS)-induced nitric oxide production in macrophage RAW264.7 cells. Myrislignan exhibited the highest inhibitory activity with an IC50 value of 19.28 μM. Following closely, compounds oleiferin F and myristargenol A displayed IC50 values of 25.56 and 26.15 μM, respectively. Compounds machilin A and nectandrin B showed moderate inhibitory activity with IC50 values of 46.56 and 41.21 μM, respectively. However, zuonin A, elemicin, eugenol, aristolignan, fragransin C1, and otobaphenol demonstrated inactivity, with IC50 values exceeding 50 μM. The results from the in vitro inhibitory activity assay prompted the application of molecular docking to explore the affinity and interactions between compounds myrislignan, oleiferin F and myristargenol A, and proteins associated with inflammation, namely inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and interleukin-8 (IL-8). Additionally, ADMET (absorption, distribution, metabolism, excretion, and toxicity) predictions were employed. These findings suggest that the active constituents from M. fragrans hold potential for further exploration and development as anti-inflammatory agents.