Drug glucuronidation assays on human liver microsomes immobilized on microfluidic flow-through reactors

Iiro Kiiski, Elisa Ollikainen, Sanna Artes, Päivi Järvinen, Ville Jokinen, Tiina Sikanen

Research output: Contribution to journalArticleScientificpeer-review

Abstract

UDP-glucuronosyltransferases (UGTs), located in the endoplasmic reticulum of liver cells, are an important family of enzymes, responsible for the biotransformation of several endogenous and exogenous chemicals, including therapeutic drugs. However, the phenomenon of 'latency', i.e., full UGT activity revealed by disruption of the microsomal membrane, poses substantial challenges for predicting drug clearance based on in vitro glucuronidation assays. This work introduces a microfluidic reactor design comprising immobilized human liver microsomes to facilitate the study of UGT-mediated drug clearance under flow-through conditions. The performance of the microreactor is characterized using glucuronidation of 8-hydroxyquinoline (via multiple UGTs) and zidovudine (via UGT2B7) as the model reactions. With the help of alamethicin and albumin effects, we show that conducting UGT metabolism assays under flow conditions facilitates in-depth mechanistic studies, which may also shed light on UGT latency.

Original languageEnglish
Article number105677
JournalEuropean Journal of Pharmaceutical Sciences
Volume158
Number of pages9
ISSN0928-0987
DOIs
Publication statusPublished - 1 Mar 2021
MoE publication typeA1 Journal article-refereed

Fields of Science

  • 116 Chemical sciences
  • drug metabolism
  • glucuronidation
  • microreactors
  • enzyme immobilization
  • microfluidics
  • microfabrication
  • HUMAN UDP-GLUCURONOSYLTRANSFERASES
  • CYTOCHROME-P450 ENZYMES
  • FATTY-ACIDS
  • METABOLISM
  • ALBUMIN
  • PHOSPHOLIPIDS
  • EXPLANATION
  • FABRICATION
  • PREDICTION
  • KINETICS
  • 317 Pharmacy
  • Drug metabolism
  • Glucuronidation
  • Microreactors
  • Enzyme immobilization
  • Microfluidics
  • Microfabrication
  • HUMAN UDP-GLUCURONOSYLTRANSFERASES
  • CYTOCHROME-P450 ENZYMES
  • FATTY-ACIDS
  • METABOLISM
  • ALBUMIN
  • PHOSPHOLIPIDS
  • EXPLANATION
  • FABRICATION
  • PREDICTION
  • KINETICS

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