Abstract

Prilocaine is a widely used local anesthetic known to induce methemoglobinemia, a hematologic disorder characterized by oxidation of hemoglobin iron from ferrous (Fe²?) to ferric (Fe³?) state, resulting in impaired oxygen transport and tissue hypoxia. Despite clinical recognition of this adverse effect, the molecular mechanisms underlying prilocaine-induced methemoglobinemia remain incompletely understood. This study aimed to investigate the toxicological mechanisms of prilocaine-induced methemoglobinemia through an integrated network toxicology and molecular docking approach. Potential molecular targets of prilocaine were identified using PubChem and SwissTargetPrediction, while methemoglobinemia-associated genes were collected from GeneCards, DisGeNET, and Ensembl databases. Overlapping targets were determined and subjected to protein–protein interaction network construction using GeneMANIA. Functional enrichment analyses, including Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis, were performed using STRING. Molecular docking was conducted using AutoDock Vina to validate interactions between prilocaine and key protein targets. Three common targets—CYP2C19, CASR, and JAK2—were identified as potential mediators of prilocaine-induced methemoglobinemia. Protein interaction network analysis highlighted these genes as central regulators linked to oxidative stress and metabolic pathways. Functional enrichment revealed significant involvement in oxidation–reduction processes, cellular oxidative stress response, heme binding, oxidoreductase activity, and xenobiotic metabolism. KEGG pathway analysis identified cytochrome P450-mediated drug metabolism, PI3K-Akt signaling, and JAK-STAT signaling as major pathways implicated in toxicity. Molecular docking demonstrated favorable binding affinity of prilocaine toward CYP2C19 (-6.9 kcal/mol), CASR (-6.8 kcal/mol), and JAK2 (-6.4 kcal/mol), supporting their functional relevance in mediating toxic effects. This study reveals that prilocaine-induced methemoglobinemia is driven by a multi-target and multi-pathway mechanism, primarily involving cytochrome P450-mediated metabolic activation, oxidative stress generation, and downstream intracellular signaling dysregulation. The identified molecular targets provide mechanistic insight into prilocaine toxicity and may serve as potential biomarkers or therapeutic targets for prevention and management of drug-induced methemoglobinemia.

Authors

Nileshraj Govindaraj¹, C. Swithraa², J.V. Sabari Anandh³, M. Tamil Selvan⁴
Arunai Medical College and Hospital, India¹, Government Tiruvannamalai Medical College and Hospital, India², St. Peter’s Medical College, Hospital and Research Institute, India³, DA Pandu Memorial RV Dental College, India⁴

Keywords

Prilocaine, Methemoglobinemia, Network Toxicology, Molecular Docking, CYP2C19, JAK2, CASR, Oxidative Stress, Cytochrome P450, Computational Toxicology