In vitro metabolism of a novel synthetic cannabinoid, EAM-2201, in human liver microsomes and human recombinant cytochrome P450s

Kim, Ju Hyun; Kim, Hee Seung; Kong, Tae Yeon; Lee, Joo Young; Kim, Jin Young; In, Moon Kyo; Lee, Hye Suk

doi:10.1016/j.jpba.2015.11.023

Cited by 23 publications

(22 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Urine case samples can be used for metabolite identification but are not entirely sufficient for this purpose because of inter‐individual variations . In addition, human liver microsomes (HLMs) and hepatocytes can be used for in vitro identification, but their application is insufficient when they are used alone . Major in vivo metabolites cannot always be predicted by in vitro methods.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Detection of metabolites of the new synthetic cannabinoid CUMYL‐4CN‐BINACA in authentic urine samples and human liver microsomes using high‐resolution mass spectrometry

Öztürk¹,

Yeter²,

Öztürk³

et al. 2017

Drug Testing and Analysis

View full text Add to dashboard Cite

CUMYL-4CN-BINACA(1-(4-cyanobutyl)-N-(2-phenylpropan-2-yl)-1H-indazole-3-carboxamide) is a recently introduced indazole-3-carboxamide-type synthetic cannabinoid (SC) that was detected in herbal incense seized by of the Council of Forensic Medicine, Istanbul Narcotics Department, in May 2016 in Turkey. Recently introduced SCs are not detected in routine toxicological analysis; therefore, analytical methods to measure these compounds are in demand. The present study aims to identify urinary marker metabolites of CUMYL-4CN-BINACA by investigating its metabolism in human liver microsomes and to confirm the results in authentic urine samples (n = 80). In this study, 5 μM CUMYL-4CN-BINACA was incubated with human liver microsomes (HLMs) for up to 3 hours, and metabolites were identified using liquid chromatography-high-resolution mass spectrometry (LC-HRMS). Less than 21% of the CUMYL-4CN-BINACA parent compound remained after 3 hours of incubation. We identified 18 metabolites that were formed via monohydroxylation, dealkylation, oxidative decyanation to aldehyde, alcohol, and carboxylic acid formation, glucuronidation or reaction combinations. CUMYL-4CN-BINACA N-butanoic acid (M16) was found to be major metabolite in HLMs. In urine samples CUMYL-4CN-BINACA was not detected; CUMYL-4CN-BINACA N-butanoic acid (M16) was major metabolite after β-glucuronidase hydrolysis. Based on these findings, we recommend using M16 (CUMYL-4CN-BINACA N-butanoic acid), M8 and M11 (hydroxylcumyl CUMYL-4CN-BINACA) as urinary marker metabolites to confirm CUMYL-4CN-BINACA intake.

show abstract

Section: Introductionmentioning

confidence: 99%

“…Major in vivo metabolites cannot always be predicted by in vitro methods. However, robust and specific metabolite identification can be achieved by combining applications . For example, in vitro identified metabolites can be confirmed by using case samples to identify major metabolites.…”

Section: Introductionmentioning

confidence: 99%

Detection of metabolites of the new synthetic cannabinoid CUMYL‐4CN‐BINACA in authentic urine samples and human liver microsomes using high‐resolution mass spectrometry

Öztürk¹,

Yeter²,

Öztürk³

et al. 2017

Drug Testing and Analysis

View full text Add to dashboard Cite

show abstract

“…Therefore, it is necessary to identify additional in vitro metabolites of MAM-2201 and separate the metabolites with the same molecular ions ([M + H] + ). To predict individual differences in synthetic cannabinoid toxicity and to avoid toxic drug-drug interactions, the drug-metabolizing enzymes of the derivatives AM-2201 and EAM-2201 were characterized using major human cytochrome P450 (CYP) enzymes [33,34]. Conversion of AM-2201 to 4-hydroxyfluoropentyl-AM-2201, AM-2201 pentanoic acid, and 5-hydroxypentyl-AM-2201 was catalyzed by CYP1A2, CYP2C9, and CYP2C19, respectively [33].…”

Section: Introductionmentioning

confidence: 99%

“…Conversion of AM-2201 to 4-hydroxyfluoropentyl-AM-2201, AM-2201 pentanoic acid, and 5-hydroxypentyl-AM-2201 was catalyzed by CYP1A2, CYP2C9, and CYP2C19, respectively [33]. Twenty-eight metabolites were formed from EAM-2201 by CYP1A2, CYP2B6, CYP2C8/9/19, CYP2D6, and CYP3A4 [34]; however, there are no reports of the CYP enzymes responsible for MAM-2201 metabolism.…”

Section: Introductionmentioning

confidence: 99%

Metabolic characterization of (1-(5-fluoropentyl)-1H-indol-3-yl)(4-methyl-1-naphthalenyl)-methanone (MAM-2201) using human liver microsomes and cDNA-overexpressed cytochrome P450 enzymes

et al. 2016

Self Cite

View full text Add to dashboard Cite

MAM-2201 is a synthetic cannabinoid that is increasingly found in recreational drug abusers and cases of severe intoxication. Thus, characterization of the metabolic pathways of MAM-2201 is necessary to predict individual pharmacokinetics and toxicity differences, and to avoid toxic drug-drug interactions. Collectively, 19 phase 1 metabolites of MAM-2201 were identified using liquid chromatography-Orbitrap mass spectrometry following human liver microsomal incubations in the presence of NADPH: 7 hydroxy-MAM-2201 (M1-M7), 4 dihydroxy-MAM-2201 (M8-M11), dihydrodiol-MAM-2201 (M12), N-(5-hydroxypentyl)-MAM-2201 (M13), hydroxy-M13 (M14), N-dealkyl-MAM-2201 (M15), 2 hydroxy-M15 (M16, M17), MAM-2201 N-pentanoic acid (M18), and hydroxy-M18 (M19). On the basis of intrinsic clearance values in human liver microsomes, hydroxy-MAM-2201 (M1), N-(5-hydroxypentyl)-MAM-2201 (M13), and hydroxy-M13 (M14) were the major metabolites. Based on an enzyme kinetics study using human cDNA-expressed cytochrome P450 (CYP) enzymes and an immunoinhibition study using selective CYP antibodies in human liver microsomes, CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 enzymes were responsible for MAM-2201 metabolism. The CYP3A4 enzyme played a prominent role in MAM-2201 metabolism, and CYP1A2, CYP2B6, CYP2C8, and CYP2C9 enzymes played major roles in the formation of some metabolites. MAM-2201 is extensively metabolized by multiple CYP enzymes, indicating that MAM-2201 and its metabolites should be used as markers of MAM-2201 abuse and toxicity. Graphical abstract In vitro metabolic pathways of MAM-2201 were characterized in human liver microsomes and recombinant CYPs using LC-HRMS analysis. Total 19 phase I metabolites were identified with predominant contribution of CYP3A4.

show abstract

“…Detection and quantification of abused drug substances in biological samples are key steps in the diagnosis and treatment of emergency cases. However, most synthetic cannabinoids are extensively metabolized, and it is therefore difficult to identify such synthetic cannabinoids in biological samples . Therefore, in vitro metabolism studies of synthetic cannabinoids using human liver microsomes and hepatocytes are performed to identify their metabolites as relevant indicators of drug abuse.…”

Section: Introductionmentioning

confidence: 99%

Targeted and non‐targeted metabolite identification of MAM‐2201 in human, mouse, and rat hepatocytes

Kim

Kong

Moon

et al. 2018

Drug Testing and Analysis

Self Cite

View full text Add to dashboard Cite

MAM-2201 is a fluorinated naphthoylindole synthetic cannabinoid with potent psychoactive properties that has been detected as an active ingredient in herbal incense blends. To gain a greater understanding of MAM-2201 metabolism and to compare its metabolic fate in humans with those in animals, the metabolism of MAM-2201 in human, mouse, and rat hepatocytes was investigated using liquid chromatography-high-resolution mass spectrometry combined with targeted and non-targeted metabolite profiling approaches. Nineteen phase I metabolites (M1-M19) reported previously in human liver microsomes and 13 novel metabolites were identified in human, mouse, and rat hepatocytes: 1 phase I metabolite (M20) and 12 phase II metabolites including 6 glucuronides (G1-G6), 1 sulfate (S1), and 5 glutathione (GSH) conjugates (GS1-GS5) of MAM-2201 metabolites. G3 was human-specific, but M20, G1, G2, and 5 GSH conjugates were rat-specific, indicating species-related differences in MAM-2201 metabolism. The findings in the present study can be useful for the experimental design and assessment of metabolism-mediated toxic risk of MAM-2201.

show abstract

In vitro metabolism of a novel synthetic cannabinoid, EAM-2201, in human liver microsomes and human recombinant cytochrome P450s

Cited by 23 publications

References 31 publications

Detection of metabolites of the new synthetic cannabinoid CUMYL‐4CN‐BINACA in authentic urine samples and human liver microsomes using high‐resolution mass spectrometry

Detection of metabolites of the new synthetic cannabinoid CUMYL‐4CN‐BINACA in authentic urine samples and human liver microsomes using high‐resolution mass spectrometry

Metabolic characterization of (1-(5-fluoropentyl)-1H-indol-3-yl)(4-methyl-1-naphthalenyl)-methanone (MAM-2201) using human liver microsomes and cDNA-overexpressed cytochrome P450 enzymes

Targeted and non‐targeted metabolite identification of MAM‐2201 in human, mouse, and rat hepatocytes

Contact Info

Product

Resources

About