Pharmacokinetic and toxicological properties of benzoic acid: integrated In silico characterization and In vitro validation with implications for dermatology

Abstract

Benzoic acid (BA) is widely used in pharmaceutical, cosmetic, and dermatological formulations, yet its pharmacokinetic and toxicological profile has not been examined within a unified predictive and experimental framework. This study aimed to characterize BA using an integrated in silico and in vitro framework with dermatological implications, combining absorption, distribution, metabolism, excretion, and toxicity (ADMET) prediction, toxicity modeling, phenotype-driven molecular docking, and HEK293 cell assays. The BA structure was retrieved from PubChem and analyzed using pkCSM and ProTox-III for ADMET prediction. Molecular docking was performed with AutoDock Vina against eight phenotype-driven target proteins representing inflammation, oxidative stress, survival, and migration pathways. In vitro validation used HEK293 cells exposed to BA at 0, 15, 250, and 1,000 μM, with cell viability, wound-healing capacity, clonogenic capacity, lipid peroxidation (malondialdehyde), catalase activity, and total oxidant/antioxidant status assessed. BA showed favorable absorption and low predicted metabolic interaction liability, with platform-dependent discrepancies between pkCSM and ProTox-III regarding hepatotoxicity and blood–brain barrier penetration. Docking yielded moderate binding affinities (−5.175 to −6.388 kcal/mol), strongest against sphingosine-1-phosphate receptor 3, catalase, and cyclooxygenase-2. In HEK293 cells, BA produced dose-dependent reductions in viability (IC50 ≈ 242 μM), migration, and colony formation, alongside dose-dependent oxidative imbalance. Overall, BA displays a favorable pharmacokinetic profile but a mixed toxicological pattern. The predictive discrepancies and dose-dependent in vitro effects indicate caution in dermatological applications of BA and support further experimental studies on its cutaneous safety.