In silico screening of Nrf2-activating phytocompounds for ischemic stroke therapy

AbstractIschemic stroke remains a major cause of death and disability worldwide, with reperfusion injury aggravating neuronal damage via oxidative stress and inflammation. Current therapies such as thrombolytics are constrained by narrow treatment windows and hemorrhagic risks, underscoring the need for alternative neuroprotective strategies. Natural compounds with antioxidant and anti-inflammatory potential, particularly those targeting the Nrf2 (nuclear factor erythroid 2-related factor 2) pathway, have emerged as promising candidates. Moringa oleifera, Phyllanthus niruri, and Nigella sativa are traditionally used medicinal plants with neuroprotective properties, yet their Nrf2-modulating potential remains insufficiently explored. This study aimed to identify and evaluate phytochemicals from these plants as potential Nrf2 modulators for ischemic stroke-induced reperfusion injury. Bioactive compounds were identified from methanolic extracts using liquid chromatography–mass spectrometry (LC-MS). Identified molecules were subjected to molecular docking against the Keap1 Kelch domain of Nrf2 to assess binding affinity, interaction profiles, and structural stability, followed by in silico ADMET (Absorption, Distribution, Metabolism, Excretion, and Toxicity) analysis to predict pharmacokinetics and safety. LC-MS profiling revealed several neuroactive phytochemicals, with Flavonol base + 4O, 1MeO, O-Hex-Hex showing the strongest binding affinity (–9.6 kcal/mol), followed by Quercetin-3,42 -O-di-ß-glucopyranoside (–9.2 kcal/mol) and Ginsenoside Rb2 (–5.3 kcal/mol). Docking analyses highlighted stable interactions with critical residues (ARG415, TYR572, GLY509, ALA556) through hydrogen bonds, p–p, and p–alkyl interactions, confirmed by 2D and 3D visualizations. ADMET predictions indicated favorable pharmacokinetic properties and low toxicity for all compounds, although Quercetin-3,4' -O-di-ß-glucopyranoside showed a potential hepatotoxicity warning. These findings suggest that Flavonol base + 4O, 1MeO, O-Hex-Hex and other phytochemicals can act as Nrf2 activators, providing mechanistic insight into their neuroprotective potential. The results support the development of plant-derived Nrf2 modulators as novel candidates for mitigating reperfusion injury in ischemic stroke. Further in vitro and in vivo studies are warranted to validate their efficacy.