Natural products, particularly terpenoids from aromatic plants like Thymus vulgaris L., 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 via 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 chromatog., and characterized by Fourier transform IR (FT-IR), NMR, and mass spectrometry.D. functional theory (DFT) at B3LYP/6-31G(d,p) optimized R1′s geometry, while mol. 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]).Mol. dynamics (MD) simulations indicated complex stability via hydrogen bonds and hydrophobic interactions.In vitro 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 in vivo studies to optimize efficacy and safety.