Molecular engineering of ω-3 PUFA-based self-assembled HTHQ prodrug nanoparticles for hepatic ischemia-reperfusion injury therapy

Liver transplantation is a definitive treatment for end-stage liver disease, but hepatic ischemia-reperfusion injury (HIRI) remains a major cause of post-transplant early graft dysfunction, with ferroptosis-associated lipid peroxidation and lipid metabolic disturbance representing key pathological features. Disease-signature-guided perturbation screening identified 1-O-hexyl-2,3,5-trimethylhydroquinone (HTHQ) as a phenolic antioxidant with anti-HIRI potential, whereas free HTHQ is limited by poor aqueous dispersibility and suboptimal hepatic exposure. Transcriptomic profiling showed that HTHQ attenuated HIRI-associated transcriptional alterations related to ferroptosis, lipid oxidation, and lipid metabolism, providing a rationale for lipid-oriented structural engineering. Accordingly, HTHQ was conjugated with eicosapentaenoic acid (EPA), an ω-3 polyunsaturated fatty acid, to generate an amphiphilic HTHQ-EPA prodrug that self-assembled into HTHQ-EPA nanoparticles (HENPs). This design integrated antioxidant activity with lipid-mediated self-assembly, resulting in improved colloidal stability, systemic exposure, and hepatic HTHQ exposure. In murine HIRI models, HENPs markedly alleviated hepatic injury, suppressed lipid peroxidation and iron accumulation, reduced inflammatory responses, and restored GSH levels and GPX4-mediated antioxidant defense. HENPs also protected hepatocytes against hypoxia/reoxygenation-induced injury and exhibited favorable biosafety in vivo. Together, our findings indicate that HENPs attenuate ferroptosis-associated hepatic injury and support lipid-engineered prodrug nanoparticles as a therapeutic approach for mitigating hepatic ischemia-reperfusion injury.