Metabolism and cytotoxicity studies of the two hallucinogens 1cP-LSD and 4-AcO-DET in human liver and zebrafish larvae models using LC-HRMS/MS and a high-content screening assay

被引：1

作者：

Gampfer, Tanja M. ^{[1
,8
]}

Schuetz, Victoria ^{[1
]}

Schippers, Philip ^{[1
,2
]}

Rasheed, Sari ^{[2
,3
]}

Baumann, Jonas ^{[2
]}

Wagmann, Lea ^{[1
]}

Pulver, Benedikt ^{[4
,5
,6
]}

Westphal, Folker ^{[4
]}

Flockerzi, Veit ^{[7
]}

Mueller, Rolf ^{[2
,3
]}

Meyer, Markus R. ^{[1
]}

机构：

[1] Saarland Univ, Inst Expt & Clin Pharmacol & Toxicol, Ctr Mol Signaling PZMS, Dept Expt & Clin Toxicol, Homburg, Germany

[2] Saarland Univ, Helmholtz Inst Pharmaceut Res Saarland HIPS, Helmholtz Ctr Infect Res HZI, Saarbrucken, Germany

[3] German Ctr Infect Res DZIF, Partner Site Hannover, Braunschweig, Germany

[4] Forens Sci Inst, State Bur Criminal Invest Schleswig Holstein, Kiel, Germany

[5] Univ Freiburg, Inst Forens Med, Fac Med, Med Ctr, Freiburg, Germany

[6] Univ Freiburg, Herrmann Staudinger Grad Sch, Freiburg, Germany

[7] Saarland Univ, Inst Expt & Clin Pharmacol & Toxicol, Ctr Mol Signaling PZMS, Dept Expt & Clin Pharmacol, Homburg, Germany

[8] Saarland Univ, Ctr Mol Signaling PZMS, Dept Expt & Clin Toxicol, D-66421 Homburg, Germany

来源：

JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS | 2024年 / 245卷

关键词：

New psychoactive substances; Pooled human liver S9 fraction; Zebrafish larvae; LC-HRMS/MS; HepG2; Cytotoxicity; EMBRYOS; DRUGS;

D O I：

10.1016/j.jpba.2024.116187

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

The continuous emergence of new psychoactive substances (NPS) attracted a great deal of attention within recent years. Lately, the two hallucinogenic NPS 1cP-LSD and 4-AcO-DET have appeared on the global market. Knowledge about their metabolism to identify potential metabolic targets for analysis and their cytotoxic properties is lacking. The aim of this work was thus to study their in vitro and in vivo metabolism in pooled human liver S9 fraction (pHLS9) and in zebrafish larvae (ZL) by means of liquid chromatography -high -resolution tandem mass spectrometry. Monooxygenases involved in the initial metabolic steps were elucidated using recombinant human isozymes. Investigations on their cytotoxicity were performed on the human hepatoma cell line HepG2 using a multiparametric, fluorescence -based high -content screening assay. This included measurement of CYP-enzyme mediated effects by means of the unspecific CYP inhibitor 1-aminbenzotriazole (ABT). Several phase I metabolites of both compounds and two phase II metabolites of 4-AcO-DET were produced in vitro and in vivo. After microinjection of 1cP-LSD into the caudal vein of ZL, three out of seven metabolites formed in pHLS9 were also detected in ZL. Twelve 4-AcO-DET metabolites were identified in ZL after exposure via immersion bath and five of them were found in pHLS9 incubations. Notably, unique metabolites of 4-AcO-DET were only produced by ZL, whereas 1cP-LSD specific metabolites were found both in ZL and in pHLS9. No toxic effects were observed for 1cP-LSD and 4-AcO-DET in HepG2 cells, however, two parameters were altered in incubations containing 4-AcO-DET together with ABT compared with incubations without ABT but in concentrations far above expected in vivo concentration. Further investigations should be done with other hepatic cell lines expressing higher levels of CYP enzymes.

引用

页数：12