In Vitro Metabolism of 25B-NBF, 2-(4-Bromo-2,5-Dimethoxyphenyl)-N-(2-Fluorobenzyl)ethanamine, in Human Hepatocytes Using Liquid Chromatography–Mass Spectrometry

Abstract

25B-NBF, 2-(4-bromo-2,5-dimethoxyphenyl)-N-(2-fluorobenzyl)ethanamine, is a new psychoactive substance classified as a phenethylamine. It is a potent agonist of the 5-hydroxytryptamine receptor, but little is known about its metabolism and elimination properties since it was discovered. To aid 25B-NBF abuse screening, the metabolic characteristics of 25B-NBF were investigated in human hepatocytes and human cDNA-expressed cytochrome P450 (CYP) and UDP-glucuronosyltransferase (UGT) enzymes using liquid chromatography–high resolution mass spectrometry. At a hepatic extraction ratio of 0.80, 25B-NBF was extensively metabolized into 33 metabolites via hydroxylation, O-demethylation, bis-O-demethylation, N-debenzylation, glucuronidation, sulfation, and acetylation after incubation with pooled human hepatocytes. The metabolism of 25B-NBF was catalyzed by CYP1A1, CYP1A2, CYP2B6, CYP2C9, CYP2C19, CYP2D6, CYP2J2, CYP3A4, and UGT2B7 enzymes. Based on these results, it is necessary to develop a bioanalytical method for the determination of not only 25B-NBF but also its metabolites in biological samples for the screening of 25B-NBF abuse.

Synthesis and Functional Characterization of 2-(2,5-Dimethoxyphenyl)-N-(2-fluorobenzyl)ethanamine (25H-NBF) Positional Isomers

“…Analogous to the NBOMe substances, O -demethylation of methoxy groups has been reported to be an important metabolic pathway in the degradation of 25X-NBF substances. At this moment, it is unknown what the impact on metabolism is of the altered positions of the methoxy groups, compared to the 2,5-dimethoxy pattern . This is one of the limitations that complicates the interpretation of data obtained in in vitro functional assays and their potential translation to in vivo outcomes.…”

“…At this moment, it is unknown what the impact on metabolism is of the altered positions of the methoxy groups, compared to the 2,5-dimethoxy pattern. 30 This is one of the limitations that complicates the interpretation of data obtained in in vitro functional assays and their potential translation to in vivo outcomes. Additionally, the presented data do not allow for the assessment of the binding affinities of the synthesized isomers, which could provide more insight on how these ligands behave in the presence of other (endogenous) ligands.…”

“…35H-NBF Imine (10e). MS (EI, 70 eV), m/z (%): 287 (M +• ), 109 (100), 136 (94), 192 (30), 287 (29), 166 (17), 137 (11), 77 (10), 108 (10), 151 (9), 91 (9). Routine Cell Culture and NanoBiT βarr2 Recruitment Assay.…”

A molecular analysis of substituted phenylethylamines as potential microtubule targeting agents through in silico methods and in vitro microtubule-polymerization activity

“…A body of work has been done on how to identify the presence of these drugs (25B-NBF, 25C-NBF, and DMBMPP) in the human body for drug detection purposes 37 – 41 , however little is known about their overall effects and mechanisms of action including potential effects on the microtubule cytoskeleton. As such, there is very limited dosage research for 25B-NBF, 25C-NBF, and DMBMPP.…”

Potential of chromatography and mass spectrometry for the differentiation of three series of positional isomers of 2‐(dimethoxyphenyl)‐N‐(2‐halogenobenzyl)ethanamines

“…Many compounds of NBOMe series as well as their metabolites were characterized by using gas chromatography with mass spectrometry (GC-MS), ultra-high performance liquid chromatography with high resolution mass spectrometry (UHPLC-HRMS) and tandem mass spectrometry (UHPLC-HRMS 2 ), nuclear magnetic resonance (NMR) spectroscopy and Fourier-transform infrared spectroscopy (FT-IR). [32][33][34][35][36][37][38] For compounds of the NBF group, a metabolic pathway is described and an analytical method for monitoring the abuse of 39 There are no data on the analysis in the scientific literature for compounds of the NBCl and NBBr groups. A behavioral activity study was published only for regioisomers of 25I-NBBr.…”