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Updated: Jan 22, 2026

Incorporating Target Protein Structure Flexibility and Dynamics in Computational Drug Discovery Using Ensemble-Based Docking Analysis
Published on: June 20, 2025
Structural determinants of oxygenated benzalkonium-like disinfectants as human and rat 5α-reductase 1 inhibitors:
Yinghao Huang1, Chentao Ding2, Shaowei Wang3
1Department of Anesthesiology and Perioperative Medicine, the Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325027, China; Key Laboratory of Pediatric Anesthesiology, Ministry of Education, Wenzhou Medical University, Wenzhou, Zhejiang 325027, China; Key Laboratory of Precision Anesthesiology of Zhejiang Province, Wenzhou Medical University, Wenzhou, Zhejiang 325027, China; Department of Gynecology and Obstetrics, the Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325027, China.
Abstract:
The widespread use of oxygenated benzalkonium-like quaternary ammonium disinfectants since COVID-19 raises concerns about their off-target effects on neurosteroidogenesis, particularly via inhibition of 5α-reductase type 1 (5α-R1), a key enzyme in neuroactive steroid synthesis. Here, we evaluated seven such compounds, identifying miramistin, methylbenzethonium, and benzethonium as potent inhibitors of human 5α-R1 (IC50 = 4.28, 5.01, and 8.80 µM, respectively), with species-dependent potency (human > rat). Mechanistic studies revealed mixed/noncompetitive inhibition, driven by binding to the NADPH site, as confirmed by surface plasmon resonance (SPR) and enzyme kinetics as well as in silico docking. Molecular docking elucidated interactions with key residues (e.g., Asn198 in human vs. Tyr199 in rat), rationalizing the observed interspecies differences, particularly for miramistin. Despite high microsomal activity, limited cell permeability diminished efficacy in intact cells, suggesting a need for cellular assay. These findings advance mechanistic understanding of 5α-R1 inhibition by these disinfectants and highlight their broader implications for potential disruption of neurosteroid pathways in humans as endocrine disruptors, regulatory risks due to species-specific pharmacodynamics, and opportunities to identify safer quaternary ammonium compounds without affecting neurosteroids. The study underscores the necessity of interspecies validation in assessments of environmental chemicals targeting steroidogenic enzymes. SYNOPSIS: QACs' potent 5α-R1 inhibition risks endocrine disruption in rodents and humans. High logP suggests bioaccumulation potential, demanding environmental fate studies.
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