Endocrine-disrupting metabolic activation of 2-nitrofluorene catalyzed by human cytochrome P450 1A1: A QM/MM approach

Nitropolycyclic aromatic hydrocarbons (NPAHs) present one of the most important airborne pollutants. Recent studies have shown that one of the most abundant NPAHs, 2-Nitrofluorene (NF), was supposed to be converted to endocrine-disrupting metabolites by cytochrome P450 1A1 (CYP1A1) in human cells. However, the mechanism is still largely unexplored. Here the metabolic activation and transformation mechanism of NF catalyzed by CYP1A1 were systematically studied with the aid of Molecular Dynamics, Density Functional Theory and Quantum Mechanics/Molecular Mechanics techniques. We evidence that CYP1A1 can activate NF through two elementary processes: (i) electrophilic addition (12.4 kcal.mol(-1)) or hydrogen abstraction (38.2 kcal.mol(-1)) and (ii) epoxidation (5.9 and 8.7 kcal.mol(-1)) or NIH shift (12.5 and 14.9 kcal.mol(-1)) or proton shuttle (12.1 kcal.mol(-1)). Electrophilic addition was found to be the rate-determining step while epoxidation rather than NIH shift or proton shuttle is the more feasible pathway after electrophilic addition. Metabolites 6,7-epoxide-2-nitrofluorene and 7,8-epoxide-2-nitrofluorene were identified as the major epoxidation products. Epoxides are unstable and easy to react with hydrated hydrogen ions and hydroxyls to produce endocrine disrupter 7-hydroxy-2-nitrofluorene. Toxic analysis shows that some of the metabolites are more toxic to model aquatic organisms (e.g. Green algea) than NF. Binding affinity analysis to human sex hormone binding globulin reveals that NF metabolites all have endocrine-disrupting potential. This study provides a comprehensive understanding on the biotransformation process of NF and may aid future studies on various NPAHs activation catalyzed by human P450 enzyme.

Automated summary

This study systematically investigates the metabolic activation and transformation mechanism of 2-Nitrofluorene (NF) by cytochrome P450 1A1 (CYP1A1). The results demonstrate that CYP1A1 can activate NF through electrophilic addition and epoxidation reactions, leading to the formation of endocrine-disrupting metabolites. The epoxides produced during the reaction are unstable and can react with hydrated hydrogen ions and hydroxyls, generating 7-hydroxy-2-nitrofluorene, an endocrine disrupter. Additionally, the NF metabolites are found to be more toxic to aquatic organisms and have endocrine-disrupting potential.