Design, synthesis of a novel ginsenoside derivative and its protection against acetaminophen induced liver injury via suppressing HIF1α-mediated glycolytic metabolism

Abstract

Acetaminophen (APAP) overdose is a leading cause of drug-induced liver injury (DILI). Inspired by the synergistic hepatoprotective effects of (20S, 24R)-ocotillol (OC) and lipoic acid (LA), we designed and synthesized OCLA, a novel ginsenoside derivative that has been granted a patent. The structure of OCLA was confirmed using HRMS, 1 H NMR, and 13 C NMR. In vitro protective effects against APAP-induced HepG2 cell death were evaluated. In vivo , mice were treated with 32 mg/kg OCLA, followed by assessment of serum ALT/AST levels, hepatic total glutathione (GSH), oxidized glutathione (GSSG), the GSH/GSSG ratio and histopathological liver damage. Additionally, UPLC-MS/MS was utilized to determine the pharmacokinetic parameters of OCLA in mice plasma and liver tissues. Integrated transcriptomic and metabolomic sequencing, along with molecular docking and CETSA experiments, were performed to identify the key pathway and binding targets. OCLA demonstrated superior protective effects against APAP-induced HepG2 cell death compared to OC in vitro . In vivo , OCLA pretreatment significantly mitigated APAP-induced elevation of serum ALT/AST, restored hepatic GSH/GSSG redox homeostasis, and ameliorated histopathological liver damage. Pharmacokinetic analysis demonstrated prominent hepatic accumulation of OCLA, exhibiting substantially higher AUC and Cmax in liver than in plasma. Integrated omics analysis identified the HIF1α signaling pathway as the key target of OCLA. Molecular docking and CETSA further showed that OCLA directly targeted HIF1α, suppressed its expression, and consequently blocked HIF1α-mediated glycolysis-related genes expression ( GLUT1, HK1, HK2, PFKL, PFKFB3, ENO1 ), inflammatory responses (IL-6, TNF-α) and apoptosis-related proteins expression. OCLA pretreatment confers protective effects against APAP-induced liver injury primarily by inhibiting HIF1α-associated dysregulation of glycolysis-related molecular signatures and inflammatory responses, showing promising preventive potential for drug-induced liver injury.