TY - JOUR
T1 - Drug glucuronidation assays on human liver microsomes immobilized on microfluidic flow-through reactors
AU - Kiiski, Iiro
AU - Ollikainen, Elisa
AU - Artes, Sanna
AU - Järvinen, Päivi
AU - Jokinen, Ville
AU - Sikanen, Tiina
PY - 2021/3/1
Y1 - 2021/3/1
N2 - 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.
AB - 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.
KW - Drug metabolism
KW - Enzyme immobilization
KW - Glucuronidation
KW - Microfabrication
KW - Microfluidics
KW - Microreactors
UR - http://www.scopus.com/inward/record.url?scp=85097791691&partnerID=8YFLogxK
U2 - 10.1016/j.ejps.2020.105677
DO - 10.1016/j.ejps.2020.105677
M3 - Article
AN - SCOPUS:85097791691
VL - 158
JO - European Journal of Pharmaceutical Sciences
JF - European Journal of Pharmaceutical Sciences
SN - 0928-0987
M1 - 105677
ER -