Synthesis, computational profiling, and anti-inflammatory potential of a thymol-mefenamic acid hybrid derivative

Natural products, particularly terpenoids from aromatic plants like <i>Thymus vulgaris L.</i>, hold significant potential in anti-inflammatory drug discovery. This study aimed to isolate bioactive terpenoids, synthesize ester derivative (R1) by reacting thymol with mefenamic acid, and evaluate its anti-inflammatory efficacy. Essential oils were extracted <i>via</i> steam distillation, and thymol (26.35%) was identified as the major component using GC-MS. R1 (2-isopropyl-5-methylphenyl 2-((2,3-dimethylphenyl)amino)benzoate) was synthesized via DCC/DMAP-mediated esterification, purified by column chromatography, and characterized by Fourier transform infrared (FT-IR), NMR, and mass spectrometry. Density functional theory (DFT) at B3LYP/6–31G(d,p) optimized R1’s geometry, while molecular docking (AutoDock 4.2.3) revealed strong binding affinities (−8.60 kcal/mol with TNF-α [PDB:2az5]; −6.20 kcal/mol with IL-6 [PDB:1alu]). Molecular dynamics (MD) simulations indicated complex stability via hydrogen bonds and hydrophobic interactions. <i>In vitro</i> assays on 3T3-L1 adipocytes demonstrated dose-dependent inhibition of pro-inflammatory cytokines: R1 (10 µg/mL) reduced TNF-α by 29.77% and IL-6 by 39.56%, though less potent than dexamethasone. Adsorption, Distribution, Metabolism, Excretion, and Toxicity (ADMET) predictions highlighted R1’s drug-likeness (Lipinski/Veber compliance) but noted hepatotoxicity and hERG inhibition risks. Spectroscopic and computational data aligned, validating R1’s structure and reactivity. These findings position R1 as a promising anti-inflammatory lead, warranting further <i>in vivo</i> studies to optimize efficacy and safety.