Computational design, synthesis, in vitro and in vivo evaluation of a 3,4-methylenedioxyphenol (sesamol) derivative as an NRF2/HO-1 pathway activator for protection against drug-induced liver injury

Abstract

Drug-induced liver injury is a major clinical and pharmacological challenge, often driven by oxidative stress and inflammation. This study aimed to develop a sesamol derivative with enhanced hepatoprotective efficacy via NRF2/HO-1 pathway activation. A focused virtual library of sesamol derivatives was generated through reaction-based enumeration and evaluated utilizing molecular docking and molecular dynamics simulations. Among 189 designed compounds, compound 133840-3CaBen (SMD) demonstrated superior binding to the KEAP1–NRF2 interface and was selected for synthesis and biological evaluation. In vitro antioxidant assays revealed that SMD exhibited a 4.5-fold lower IC₅₀ compared to sesamol. In HepG2 cells, SMD conferred up to 62.99% protection against paracetamol-induced toxicity. In vivo, SMD (100 & 200 mg per kg b.w. orally) significantly restored liver enzyme profiles, and antioxidant markers (CAT, GSH, GPx, SOD) and reduced MDA levels. ELISA analysis confirmed NRF2, HO-1, and γ-GCS upregulation with a concomitant decrease in TNF-α and IL-6. Histopathological examination of H&E-stained liver sections corroborated these findings, showing preserved hepatic architecture with minimal necrosis, inflammation, and vacuolation, especially in the SMD group, comparable to silymarin. Collectively, our findings suggest that structural modification of sesamol into SMD confers hepatoprotection, likely through upregulation of NRF2/HO-1 pathway proteins. These findings highlight SMD as a promising lead compound for developing NRF2-targeted hepatoprotective agents.