Use of Phenotypically Poor Metabolizer Individual Donor Human Liver Microsomes To Identify Selective Substrates of UGT2B10

Milani, Nicoló; Qiu, NaHong; Molitor, Birgit; Badée, Justine; Cruciani, Gabriele; Fowler, Stephen

doi:10.1124/dmd.119.089482

Cited by 7 publications

(7 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Further in vitro assays must be performed in order to confirm in silico predictions and test the validity of their results. Developed enzymatic assays using recombinant UGT2B10 coupled with liquid chromatography-tandem mass spectrometry analytical methods [ 20 , 57 , 81 , 82 ] could be implemented to screen our test compounds as substrates or inhibitors of UGT2B10.…”

Section: Discussionmentioning

confidence: 99%

A potential implication of UDP-glucuronosyltransferase 2B10 in the detoxification of drugs used in pediatric hematopoietic stem cell transplantation setting: an in silico investigation

Robin

Hassine

Muthukumaran

et al. 2022

BMC Mol and Cell Biol

View full text Add to dashboard Cite

Background Sinusoidal occlusion syndrome (SOS) is a potentially severe complication following hematopoietic stem cell transplantation (HSCT) in pediatric patients. Treatment related risk factors such as intensity of conditioning, hepatotoxic co-medication and patient related factors such as genetic variants predispose individuals to develop SOS. The variant allele for SNP rs17146905 in UDP-glucuronosyl transferase 2B10 (UGT2B10) gene was correlated with the occurrence of SOS in an exome-wide association study. UGT2B10 is a phase II drug metabolizing enzyme involved in the N-glucuronidation of tertiary amine containing drugs. Methods To shed light on the functionality of UGT2B10 enzyme in the metabolism of drugs used in pediatric HSCT setting, we performed in silico screening against custom based library of putative ligands. First, a list of potential substrates for in silico analysis was prepared using a systematic consensus-based strategy. The list comprised of drugs and their metabolites used in pediatric HSCT setting. The three-dimensional structure of UGT2B10 was not available from the Research Collaboratory Structural Bioinformatics - Protein Data Bank (RCSB - PDB) repository and thus we predicted the first human UGT2B10 3D model by using multiple template homology modeling with MODELLER Version 9.2 and molecular docking calculations with AutoDock Vina Version 1.2 were implemented to quantify the estimated binding affinity between selected putative substrates or ligands and UGT2B10. Finally, we performed molecular dynamics simulations using GROMACS Version 5.1.4 to confirm the potential UGT2B10 ligands prioritized after molecular docking (exhibiting negative free binding energy). Results Four potential ligands for UGT2B10 namely acetaminophen, lorazepam, mycophenolic acid and voriconazole n-oxide intermediate were identified. Other metabolites of voriconazole satisfied the criteria of being possible ligands of UGT2B10. Except for bilirubin and 4-Hydroxy Voriconazole, all the ligands (particularly voriconazole and hydroxy voriconazole) are oriented in substrate binding site close to the co-factor UDP (mean ± SD; 0.72 ± 0.33 nm). Further in vitro screening of the putative ligands prioritized by in silico pipeline is warranted to understand the nature of the ligands either as inhibitors or substrates of UGT2B10. Conclusions These results may indicate the clinical and pharmacological relevance UGT2B10 in pediatric HSCT setting. With this systematic computational methodology, we provide a rational-, time-, and cost-effective way to identify and prioritize the interesting putative substrates or inhibitors of UGT2B10 for further testing in in vitro experiments.

show abstract

Section: Discussionmentioning

confidence: 99%

A potential implication of UDP-glucuronosyltransferase 2B10 in the detoxification of drugs used in pediatric hematopoietic stem cell transplantation setting: an in silico investigation

Robin

Hassine

Muthukumaran

et al. 2022

BMC Mol and Cell Biol

View full text Add to dashboard Cite

show abstract

“…Some contribution from UGT2B11 may have added an additional signal into the proteomics measurement, reducing the abundance-activity correlation. Anecdotally, the poor metabolizer phenotype donor H0295 (Milani et al, 2020) gave an extremely low but non-zero UGT2B10 abundance signal which may be indicative of background ANV…K signal coming from other sources in liver microsomes.…”

Section: Discussionmentioning

confidence: 99%

“…Multiple substrates were tested for UGT2B7 and they showed no correlation except zidovudine, which showed a moderate correlation (rs = 0.34). Amitryptyline activities showed moderate correlation with UGT2B10 expression (rs = 0.31) whereas the more selective substrate RO5263397 (Milani et al, 2020) showed higher correlation (r=0.53), and S-oxazepam activities showed low correlation with UGT2B15 expression (rs = 0.28). UGT2B17 abundance showed higher correlation with testosterone glucuronidation (rs = 0.58), potentially benefitting from a wider dynamic range of enzyme expression.…”

Section: Correlation Of Ugt Expressionmentioning

confidence: 99%

Characterization of Hepatic UDP-Glucuronosyltransferase Enzyme Abundance-Activity Correlations and Population Variability Using a Proteomics Approach and Comparison with Cytochrome P450 Enzymes

et al. 2021

Self Cite

View full text Add to dashboard Cite

The expression of ten major drug-metabolizing UDP-glucuronosyl transferase (UGT) enzymes in a panel of 130 human hepatic microsomal samples was measured using a LC-MS/MS-based approach. Simultaneously, ten cytochrome P450s and P450 reductase were also measured and activity-expression relationships assessed for comparison. The resulting data sets demonstrated that, with the exception of UGT2B17, 10 th -90 th percentiles of UGT expression spanned 3-to 8fold ranges. These ranges were small relative to ranges of reported mean UGT enzyme expression across different laboratories. We tested correlation of UGT expression with enzymatic activities using selective probe substrates. A high degree of abundance-activity correlation (rs > 0.6) was observed for UGT1As (1A1, 3, 4, 6) and CYPs. In contrast, protein abundance and activity did not correlate strongly for UGT1A9 and UGT2B enzymes (2B4, 7, 10, 15 and 17). Protein abundance was strongly correlated for UGTs 2B7, 2B10 and 2B15. We suggest a number of factors may contribute to these differences including incomplete selectivity of probe substrates, correlated expression of these UGT2B isoforms, and the impact of splice and This article has not been copyedited and formatted. The final version may differ from this version.

show abstract

“…Kaivosaari et al (Kaivosaari et al, 2008) reported the involvement of UGT2B10 as the high affinity enzyme mainly involved in the metabolism of both levomedetomidine and dexmedetomidine but with higher intrinsic clearance of levomedetomidine. Dexmedetomidine is the active drug moiety and is also a highly selective UGT2B10 substrate (Milani et al, 2020), even though turnover was significantly less than that observed for levomedetomidine (Supplemental Figure 1). From the reported HLM kinetics data, glucuronidation intrinsic clearance values of ~150 and ~25 µL/min/mg were estimated for levomedetomidine and dexmedetomidine, respectively, which were in close agreement with values from this study.…”

Section: Medetomidinementioning

confidence: 96%

“…In vivo and in vitro studies showed that the pure enantiomer RO5263397 was cleared via UGT2B10 glucuronidation with a minor contribution from UGT1A4 (Fowler et al, 2015). In subsequent experiments RO5263397 was found to be the most selective of known UGT2B10 metabolized drugs when considering a combination of CYP and UGT-mediated metabolism processes (Milani et al, 2020). The observation of significant stereoselectivity differences in the intrinsic clearance of RO5263397 and its enantiomer RO5263396 raised the question of whether these in vitro effects would have in vivo relevance.…”

Section: Introductionmentioning

confidence: 99%

Contribution of UGT Enzymes to Human Drug Metabolism Stereoselectivity: A Case Study of Medetomidine, RO5263397, Propranolol, and Testosterone

Milani¹,

Qiu

Fowler

2022

Drug Metab Dispos

View full text Add to dashboard Cite

The enantiomeric forms of chiral compounds have identical physical properties but may vary greatly in their metabolism by individual enzymes. Enantioselectivity in UDP-glucuronosyl transferase (UGT) metabolism has been reported for a number of compounds and with different UGT isoforms involved. However, the impact of such individual enzyme results on overall clearance stereoselectivity is often not clear. The enantiomers of medetomidine, RO5263397, and propranolol and the epimers testosterone and epitestosterone exhibit more than 10-fold difference in glucuronidation rates by individual UGT enzymes. In this study we examined the translation of human UGT stereoselectivity to hepatic drug clearance considering the combination of multiple UGTs to overall glucuronidation, the contribution of other metabolic enzymes such as cytochromes P450, and the potential for differences in protein binding and blood/plasma partitioning. For medetomidine and RO5263397, the high individual enzyme (UGT2B10) enantioselectivity translated into ~3-to >10-fold differences in predicted human hepatic in vivo clearance. For propranolol the UGT enantioselectivity was irrelevant in the context of high cytochrome P450 (CYP) metabolism. For testosterone a complex picture emerged, due to differential epimeric selectivity of various contributing enzymes and potential for extrahepatic metabolism. Quite different patterns of CYP and UGT-mediated metabolism were observed across species, as well as differences in stereoselectivity, indicating that extrapolation from human enzyme and tissue data is essential when predicting human clearance enantioselectivity.

show abstract

Use of Phenotypically Poor Metabolizer Individual Donor Human Liver Microsomes To Identify Selective Substrates of UGT2B10

Cited by 7 publications

References 30 publications

A potential implication of UDP-glucuronosyltransferase 2B10 in the detoxification of drugs used in pediatric hematopoietic stem cell transplantation setting: an in silico investigation

A potential implication of UDP-glucuronosyltransferase 2B10 in the detoxification of drugs used in pediatric hematopoietic stem cell transplantation setting: an in silico investigation

Characterization of Hepatic UDP-Glucuronosyltransferase Enzyme Abundance-Activity Correlations and Population Variability Using a Proteomics Approach and Comparison with Cytochrome P450 Enzymes

Contribution of UGT Enzymes to Human Drug Metabolism Stereoselectivity: A Case Study of Medetomidine, RO5263397, Propranolol, and Testosterone

Contact Info

Product

Resources

About