LC–MS/MS method for the simultaneous quantification of intestinal CYP and UGT activity

Busch, Diana; Fritz, Anja; Partecke, Lars Ivo; Heidecke, Claus–Dieter; Oswald, Stefan

doi:10.1016/j.jpba.2018.04.003

Cited by 11 publications

(12 citation statements)

References 23 publications

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“…For example, UGT2B7, the main metabolising enzyme of naloxone, is expressed in significant amounts in kidney 24 and intestine. 25,26 This high clearance is consistent with our observation that naloxone plasma concentrations decreased rapidly after discontinuation of the infusion, whereas naloxone plasma concentrations were observable for the entire observation period, mostly because of its re-distribution kinetics. N3G pharmacokinetics were adequately described using a two-compartment model.…”

Section: Discussionsupporting

confidence: 90%

High-dose naloxone, an experimental tool uncovering latent sensitisation: pharmacokinetics in humans

Papathanasiou

Springborg

Kongstad

et al. 2019

British Journal of Anaesthesia

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Background: Naloxone, an opioid receptor antagonist, is used as a pharmacological tool to detect tonic endogenous activation of opioid receptors in experimental pain models. We describe a pharmacokinetic model linking naloxone pharmacokinetics to its main metabolite after high-dose naloxone infusion. Methods: Eight healthy volunteers received a three-stage stepwise high-dose i.v. naloxone infusion (total dose 3.25 mg kg À1). Naloxone and naloxone-3-glucuronide (N3G) plasma concentrations were sampled from infusion onset to 334 min after infusion discontinuation. Pharmacokinetic analysis was performed using non-linear mixed effect models (NONMEM). The predictive performances of Dowling's and Yassen's models were evaluated, and target-controlled infusion simulations were performed. Results: Three-and two-compartment disposition models with linear elimination kinetics described the naloxone and N3G concentration time-courses, respectively. Two covariate models were developed: simple (weight proportional) and complex (with the shallow peripheral volume of distribution linearly increasing with body weight). The median prediction error (MDPE) and wobble for Dowling's model were e32.5% and 33.4%, respectively. For Yassen's model, the MDPE and wobble were 1.2% and 19.9%, respectively. Conclusions: A parentemetabolite pharmacokinetic model was developed for naloxone and N3G after high-dose naloxone infusion. No saturable pharmacokinetics were observed. Whereas Dowling's model was inaccurate and over-predicted naloxone concentrations, Yassen's model accurately predicted naloxone pharmacokinetics. The newly developed covariate models may be used for high-dose TCI-naloxone for experimental and clinical practice. Clinical trials registration: NCT01992146.

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Section: Discussionsupporting

confidence: 90%

High-dose naloxone, an experimental tool uncovering latent sensitisation: pharmacokinetics in humans

Papathanasiou

Springborg

Kongstad

et al. 2019

British Journal of Anaesthesia

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“…By using advanced analytical methods, it has been demonstrated that the majority of drug metabolism enzymes can be detected in intestinal microsomes. Moreover, it has been shown that microsomes can be used to investigate enzymatic activity and the intestinal contribution to first-pass drug metabolism (Gröer et al, 2014;Nakamura et al, 2016;Busch et al, 2018). However, microsomes lack membrane transporters as well as low abundant enzyme systems, and the enzymatic activity in microsomes is highly dependent on the isolation method (Emoto et al, 2000a;van de Kerkhof et al, 2005;Peters et al, 2016;Drozdzik et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

Regional Differences in Human Intestinal Drug Metabolism

et al. 2018

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The intestines are key for the absorption of nutrients and water as well as drug metabolism, and it is well known that there are clear differences in the expression profile of drug metabolism enzymes along the intestinal tract. Yet only a few studies have thoroughly investigated regional differences in human intestinal drug metabolism. In this study, we evaluated phase I and phase II metabolism in matched human ileum and colon precision-cut intestinal slices (PCIS). To this end, human PCIS were incubated for 3 hours with testosterone and 7-hydroxycoumarin (7-HC) to examine phase I and phase II metabolism, respectively. Metabolite formation was assessed by high-performance liquid chromatography analysis.Our results demonstrated that androstenedione, 6b-hydroxytestosterone, 2b-hydroxytestosterone, and 7-HC sulfate were predominantly formed in the ileum, while 15a-hydroxytestosterone and 7-HC glucuronide were mainly produced in the colon. Moreover, we also observed sex differences in phase II metabolite formation, which appeared to be higher in men compared with women. Taken together, we demonstrated that phase I metabolism predominantly occurs in ileum PCIS, while phase II metabolism mostly takes place in colon PCIS. Moreover, we revealed that human PCIS can be used to study both regional and sex differences in intestinal metabolism. This work was supported by De Nederlandse organisatie voor gezondheidsonderzoek en zorginnovatie (ZonMW)-The Netherlands [Grant 114025003]. https://doi.org/10.1124/dmd.118.083428.s This article has supplemental material available at dmd.aspetjournals.org.

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“…Notably, the glucuronidation site of denopamine in human is the alcoholic hydroxyl group, but not the phenolic group . Naloxone is frequently used as a probe substrate for UGT2B7, while only kinetic data for naloxone‐3‐O‐glucuronidation in pooled human jejunal microsomes was reported by Diana et al Chloramphenicol, a broad spectrum antibiotic, has been reported to be glucuronidated at the 3‐O‐ and 1‐O alcoholic hydroxyl group predominantly by UGT2B7 in HLM with minor contributions from UGT1A6 and UGT1A9 (Table S6, Supporting Information) . 3‐Epideacetycinobufagin‐3‐O‐glucuronidation has been reported to be a high selective probe reaction for UGT2B7, which displays high isoform specificity and obeys Michaelis‐Menten kinetics with a low K m value (Table S6, Supporting Information) .…”

Section: Non‐fluorescent Probes For Human Ugtsmentioning

confidence: 99%

Chemical Probes for Human UDP‐Glucuronosyltransferases: A Comprehensive Review

Zhang

Hou

et al. 2018

Biotechnology Journal

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UGTs play crucial roles in the metabolism and detoxification of both endogenous and xenobiotic compounds. The key roles of UGTs in human health have garnered great interest in the design and development of specific probes for human UGTs. However, in contrast to other human enzymes, the probe substrates for human UGTs are rarely reported, owing to the highly overlapping substrate specificities of UGTs and the lack of the integrated crystal structures of UGTs. Over the past decades, many efforts are made to develop specific probe substrates for UGTs and use them in both basic research and drug discovery. This review focuses on recent progress in the development of probe substrates for UGTs and their biomedical applications. A long list of chemical probes for UGTs, including non-fluorescent and fluorescent probes along with their structural information and kinetic parameters, are prepared and analyzed. Additionally, challenges and future directions in this field are highlighted in the final section. All information and knowledge presented in this review provide practical tools/methods for measuring UGT activities in complex biological samples, which will be very helpful for rapid screening and characterization of UGT modulators, and for exploring the relevance of UGT enzymes to human diseases.

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LC–MS/MS method for the simultaneous quantification of intestinal CYP and UGT activity

Cited by 11 publications

References 23 publications

High-dose naloxone, an experimental tool uncovering latent sensitisation: pharmacokinetics in humans

High-dose naloxone, an experimental tool uncovering latent sensitisation: pharmacokinetics in humans

Regional Differences in Human Intestinal Drug Metabolism

Chemical Probes for Human UDP‐Glucuronosyltransferases: A Comprehensive Review

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