Metabolic Profile of Synthetic Cannabinoids 5F-PB-22, PB-22, XLR-11 and UR-144 by Cunninghamella elegans

Abstract

The knowledge of metabolic profile of synthetic cannabinoids is important for the detection of the drugs in urinalysis due to the typical absence or low abundance of parent cannabinoids in human urine. The fungus Cunninghamella elegans has been reported to be a useful tool for metabolism study and thus applicability to synthetic cannabinoids metabolism was examined. In this study, 8-quinolinyl 1-(5-fluoropentyl)-1H-indole-3-carboxylate (5F-PB-22), 8-quinolinyl 1-pentyl-1H-indole-3-carboxylate (PB-22), [1-(5-fluoropentyl)-1H-indol-3-yl](2,2,3,3-tetramethylcyclopropyl)methanone (XLR-11) and (1-pentyl-1H-indol-3-yl)(2,2,3,3-tetramethylcyclopropyl)methanone (UR-144) were incubated with Cunninghamella elegans and the metabolites were identified using liquid chromatographyquadrupole time-of-flight mass spectrometry. The obtained metabolites were compared with reported human metabolites to assess the suitability of the fungus to extrapolate human metabolism. 5F-PB-22 underwent, dihydroxylation, dihydrodiol formation, oxidative defluorination, oxidative defluorination to carboxylic acid, ester hydrolysis and glucosidation, alone and/or in combination. The metabolites of PB-22 were generated by hydroxylation, dihydroxylation, trihydroxylation, dihydrodiol formation, ketone formation, carboxylation, ester hydrolysis and glucosidation, alone and/or in combination. XLR-11 was transformed through hydroxylation, dihydroxylation, aldehyde formation, carboxylation, oxidative defluorination, oxidative defluorination to carboxylic acid and glucosidation, alone and/or in combination. UR-144 was metabolised by hydroxylation, dihydroxylation, trihydroxylation, aldehyde formation, ketone formation, carboxylation, N-dealkylation and combinations. These findings were consistent with previously reported human metabolism except for the small extent of ester hydrolysis observed and absence of glucuronidation. Despite the limitations, Cunninghamella elegans demonstrated the capacity to produce a wide variety of metabolites including some major human metabolites of XLR-11 and UR-144 at high abundance, showing the potential for metabolism of newly emerging synthetic cannabinoids.

Current applications of high-resolution mass spectrometry for the analysis of new psychoactive substances: a critical review

“…Once a component of interest has been selected, a putative structure can be postulated from the molecular formula, DBE and interpretation of MS/MS spectra using known CID pathways [82,93,95]. However, the assessment of MS/MS spectra requires experienced analysts who are familiar with the CID pathways of NPS.…”

In vitro phase I metabolism of three phenethylamines 25D‐NBOMe, 25E‐NBOMe and 25N‐NBOMe using microsomal and microbial models

“…30 This class of fungi has the ability to facilitate reactions such as hydroxylation, carboxylation, dihydrodiol formation, oxidative defluorination, Ndealkylation, glucuronidation and sulfation, 31,32 as well as those catalyzed by human CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4. 29,[32][33][34] It was recently shown by Nielsen et al 35 The aim of the present study was to conduct metabolism studies of three different NBOMe analogs -25D-NBOMe, 25E-NBOMe, and 25N-NBOMeemploying pHLM and C. elegans and to identify phase I metabolites based on mass spectrometric data. Besides the most common analogs 25B-NBOMe and 25I-NBOMe, the three analogs have previously been identified on confiscated blotter papers.…”

“…Moreover , C. elegans has the enzymatic activity of both phase I and II enzymes and holds the cytochrome P450 CYP509A1 enzyme . This class of fungi has the ability to facilitate reactions such as hydroxylation, carboxylation, dihydrodiol formation, oxidative defluorination, N‐ dealkylation, glucuronidation and sulfation, as well as those catalyzed by human CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 . It was recently shown by Nielsen et al that the main enzymes involved in the 25I‐NBOMe metabolism are CYP3A4 and CYP2D6.…”

Tentative identification of the metabolites of (1‐(cyclohexylmethyl)‐1H‐indol‐3‐yl)‐(2,2,3,3‐tetramethylcyclopropyl)methanone, and the product of its thermal degradation, by in vitro and in vivo methods

“…In vitro human liver hepatocytes or microsomes and in vivo human methodologies have been employed to study the metabolic profiles of the parent compounds and degradants. Incubation of UR‐144 and XLR‐11 with the fungus, Cunninghamella elegans , was applied to obtain sufficient amounts of metabolites for the structural identification by nuclear magnetic resonance (NMR) spectroscopy . The major metabolic pathways of UR‐144 and XLR‐11 identified mono‐dihydroxylation and carboxylation of TMCP moiety and combinations of these processes.…”