From street to lab: in vitro hepatotoxicity of buphedrone, butylone and 3,4-DMMC

Bravo, Rita Roque; Carmo, Helena; Valente, Maria João; Silva, João Pedro; Carvalho, Félix; Bastos, Maria de Lourdes; Silva, Diana Dias da

doi:10.1007/s00204-021-02990-9

Cited by 6 publications

(5 citation statements)

References 78 publications

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“…The hepatotoxicity of NPS may be mediated not only by the parent compounds but also by their metabolites, thus the metabolism-related impact was investigated using the broad CYP inhibitor ABT (Dias da Silva et al 2019;Roque Bravo et al 2021). Based on the current findings of the isozyme mapping, various isozymes were involved in the SC metabolism and the use of single isozyme inhibitors appeared not to be appropriate.…”

Section: Metabolism-based Effects On the Cytotoxicitymentioning

confidence: 99%

“…Furthermore, a recent study revealed considerable differences between HepaRG and primary human hepatocytes among others with respect to CYP enzymes and transporter levels (Hammer et al 2021). Alternatively, primary rat hepatocytes have been used to examine metabolism mediated effects (Dias da Silva et al 2019;Roque Bravo et al 2021) but the predictions of human toxicity deduced from mammalian cells have been reported to be lower than human cell lines in HCSA (O'Brien and Edvardsson 2017).…”

Section: Metabolism-based Effects On the Cytotoxicitymentioning

confidence: 99%

“…Some reports associated their intake with liver failure, but knowledge about their hepatotoxic potential is sparse or even unknown (Solimini et al 2017). Several in vitro strategies to assess the hepatotoxicity have been published using the hepatoma cell lines HepG2 or HepaRG, primary human hepatocytes, or rat hepatocytes (Dias da Silva et al 2019;Luethi et al 2017;O'Brien and Edvardsson 2017;Richter et al 2019a;Roque Bravo et al 2021). Most of them tested single cytotoxicity biomarkers in individual experiments, e.g., cell viability, collapse of mitochondrial membrane potential, increase of reactive oxygen or nitrogen species, damage on mitochondrial redox activity, leakage of lactate dehydrogenase (LDH) or cell death (Dias da Silva et al 2019;Luethi et al 2017;Roque Bravo et al 2021).…”

Section: Introductionmentioning

confidence: 99%

“…Several in vitro strategies to assess the hepatotoxicity have been published using the hepatoma cell lines HepG2 or HepaRG, primary human hepatocytes, or rat hepatocytes (Dias da Silva et al 2019;Luethi et al 2017;O'Brien and Edvardsson 2017;Richter et al 2019a;Roque Bravo et al 2021). Most of them tested single cytotoxicity biomarkers in individual experiments, e.g., cell viability, collapse of mitochondrial membrane potential, increase of reactive oxygen or nitrogen species, damage on mitochondrial redox activity, leakage of lactate dehydrogenase (LDH) or cell death (Dias da Silva et al 2019;Luethi et al 2017;Roque Bravo et al 2021). To prevent false-negative or -positive outcomes and to gain a deeper insight into intracellular processes, it is favorable to monitor multiple parameters at a subcellular level within the same experiment using high-content screening assays (HCSA) (O'Brien and Edvardsson 2017;Richter et al 2019a).…”

Section: Introductionmentioning

confidence: 99%

“…1. As not only parent compounds may cause hepatotoxic effects but also their metabolites (Dias da Silva et al 2019;Roque Bravo et al 2021), metabolism-based effects were also assessed. Moreover, the phase I metabolism of the SC was investigated, as knowledge about screening targets, crucial in analytical toxicology, is limited.…”

Section: Introductionmentioning

confidence: 99%

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Cytotoxicity, metabolism, and isozyme mapping of the synthetic cannabinoids JWH-200, A-796260, and 5F-EMB-PINACA studied by means of in vitro systems

et al. 2021

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Intake of synthetic cannabinoids (SC), one of the largest classes of new psychoactive substances, was reported to be associated with acute liver damage but information about their hepatotoxic potential is limited. The current study aimed to analyze the hepatotoxicity including the metabolism-related impact of JWH-200, A-796260, and 5F-EMB-PINACA in HepG2 cells allowing a tentative assessment of different SC subclasses. A formerly adopted high-content screening assay (HCSA) was optimized using a fully automated epifluorescence microscope. Metabolism-mediated effects in the HCSA were additionally investigated using the broad CYP inhibitor 1-aminobenzotriazole. Furthermore, phase I metabolites and isozymes involved were identified by in vitro assays and liquid chromatography–high-resolution tandem mass spectrometry. A strong cytotoxic potential was observed for the naphthoylindole SC JWH-200 and the tetramethylcyclopropanoylindole compound A-796260, whereas the indazole carboxamide SC 5F-EMB-PINACA showed moderate effects. Numerous metabolites, which can serve as analytical targets in urine screening procedures, were identified in pooled human liver microsomes. Most abundant metabolites of JWH-200 were formed by N-dealkylation, oxidative morpholine cleavage, and oxidative morpholine opening. In case of A-796260, most abundant metabolites included an oxidative morpholine cleavage, oxidative morpholine opening, hydroxylation, and dihydroxylation followed by dehydrogenation. Most abundant 5F-EMB-PINACA metabolites were generated by ester hydrolysis plus additional steps such as oxidative defluorination and hydroxylation. To conclude, the data showed that a hepatotoxicity of the investigated SC cannot be excluded, that metabolism seems to play a minor role in the observed effects, and that the extensive phase I metabolism is mediated by several isozymes making interaction unlikely.

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Section: Metabolism-based Effects On the Cytotoxicitymentioning

confidence: 99%