Multiscale Computational Investigations of Keap1 Inhibitors for Kidney Disease Treatments: Integrating in silico and in vivo Approaches

Kidney injury, a growing global health concern, lacks effective therapeutic interventions. This study aimed to identify natural compounds from Geophila obvallata (Gob) that modulate the Keap1/Nrf2 pathway, a key mediator of cadmium-induced nephrotoxicity. Eighty-five bioactive compounds from Gob were screened via molecular docking, quantitative structure-activity relationship (QSAR) modeling, and absorption, distribution, metabolism, excretion, toxicity (ADMET) profiling against Keap1. The top candidates, Quercetin-3-rhamnoside (Q3R) and Narcissin, along with the reference compound Resveratrol, were further evaluated using 100-ns molecular dynamics (MD) simulations. The Q3R-Keap1 complex demonstrated superior conformational stability (RMSD: 1–3 Å), outperforming Resveratrol. To validate these computational insights, an in vivo study was conducted with 28 male mice divided into four groups: control, Cd-exposed (0.3 mg/kg), Cd + Q3R (0.3 mg/kg Cd + 50 mg/kg Q3R), and Q3R alone (50 mg/kg). Treatments were administered via oral gavage for 28 days. Cadmium exposure significantly increased blood urea nitrogen (BUN) and creatinine levels, elevated reactive oxygen species (ROS: 9.93 U/mg tissue) and malondialdehyde (MDA: 5.03 nmol/mg protein), and suppressed antioxidant enzymatic activities (GPx, GSH, GST, CAT, SOD). Q3R co-administration significantly attenuated cadmium-induced renal dysfunction BUN (46.78 mg/dL vs. 72.24 mg/dL; creatinine: 1.03 vs. 2.48 mg/dL), reversed oxidative stress markers, and restored antioxidant enzyme activity. These findings demonstrate the potential of integrated computational and natural product approaches in developing novel therapies for kidney diseases.