Antioxidant and Anti-Inflammatory Constituents from the Roots of Anodendron affine: Inhibition of the fMLP-Induced Superoxide Anion Generation and Molecular Docking Studies

Oxidative stress is a key driver of chronic inflammatory diseases. <i>Anodendron affine</i> is a native Formosan plant species in Taiwan that remains largely underexplored phytochemically and bioactivity. To reveal the bioactive constituents and assess its potential as a source of anti-inflammatory antioxidants, we performed bioactivity-guided fractionation and evaluated the inhibition of superoxide anion (O₂^•-) generation in formyl-L-methionyl-L-leucyl-L-phenylalanine-stimulated human neutrophils. Molecular docking simulations were employed to model interactions with Formyl peptide receptor 1 (FPR1) and the Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase complex, including neutrophil cytosol factor 1 (p47phox) and NADPH oxidase 2 (NOX2), to propose a theoretical mechanism of action. Phytochemical investigation led to the isolation of two new compounds, methyl 4,5-<i>O</i>-diferuloyl-3-methoxyquinate (<b>1</b>) and 16-pregnen-3,12,20-trione (<b>2</b>), together with four known compounds. Notably, 4-hydroxy-3-prenylbenzoic acid (<b>5</b>) exhibited potent inhibitory activity (IC₅₀ = 17.65 ± 0.97 μM), surpassing the activity of the positive control, ibuprofen (IC₅₀ = 27.85 ± 3.56 μM). Docking studies suggested that anodendrosin H (<b>4</b>) and 4-hydroxy-3-prenylbenzoic acid (<b>5</b>) exhibit high predicted binding affinity to p47phox and NOX2. Based on these results, compounds <b>1</b>, <b>4</b>, and <b>5</b> from <i>A. affine</i> were identified as potential lead candidates for the development of novel anti-inflammatory therapeutics.