Metabolism of highly potent synthetic opioid nitazene analogs: N‐ethyl‐N‐(1‐glucuronyloxyethyl) metabolite formation and degradation to N‐desethyl metabolites during enzymatic hydrolysis

Abstract The metabolism of the highly potent synthetic opioids metonitazene, etonitazene, and protonitazene was investigated in fresh human hepatocytes. In the hydrolyzed culture medium, N ‐desethyl‐, N , N ‐di‐desethyl‐, O ‐desalkyl‐, N ‐desethyl‐ O ‐desalkyl‐, N , N ‐di‐desethyl‐ O ‐desalkyl‐, and N ‐oxidated metabolites were detected as phase I metabolites, whereas in the unhydrolyzed culture medium, O ‐glucuronides of phase I metabolites with O ‐dealkylation were detected as phase II metabolites. The detected phase I metabolites were identified by comparing their analytical data with those of synthesized authentic standards. In contrast, phase II metabolites were identified by comparing their analytical data with those of the glucuronidated products formed by the incubation of the corresponding substrates with human liver microsomes in the presence of uridine diphosphate glucuronic acid. In addition to the aforementioned metabolites, some putative N ‐ethyl‐ N ‐(1‐glucuronyloxyethyl) metabolites were detected in the unhydrolyzed culture medium. Purification and hydrolysis experiments revealed that N ‐ethyl‐ N ‐(1‐glucuronyloxyethyl) metabolites formed the corresponding N ‐desethyl metabolites via unstable N ‐ethyl‐ N ‐(1‐hydroxyethyl) metabolites during enzymatic hydrolysis.