Primary Human Hepatocyte Spheroids as an In Vitro Tool for Investigating Drug Compounds with Low Hepatic Clearance

Abstract

Characterizing the pharmacokinetic properties of drug candidates represents an essential task during drug development. In the past, liver microsomes and primary suspended hepatocytes have been extensively used for this purpose, but their relatively short stability limits the applicability of such in vitro systems for drug compounds with low metabolic turnover. In the present study, we used 3D primary human hepatocyte spheroids to predict the hepatic clearance of seven drugs with low to intermediate clearance in humans. Our results indicate that hepatocyte spheroids maintain their in vivo like phenotype during prolonged incubations allowing to monitor the depletion of parent drug for seven days. In This article has not been copyedited and formatted. The final version may differ from this version.

Organotypic and Microphysiological Human Tissue Models for Drug Discovery and Development—Current State-of-the-Art and Future Perspectives

“…In an industryled study using a small panel of four low-to-intermediate clearance compounds, intrinsic clearance could be predicted within 3-fold using as little as 6000 hepatocytes (Kanebratt et al, 2021). In agreement with these findings, clearance of a nonoverlapping set of seven lowand intermediate-clearance compounds was accurately predicted with an average fold error of 0.7 (Riede et al, 2021). Irrespective of the model, the turnover of highclearance compounds tended to be underpredicted, whereas low-clearance compound predictions were more accurate.…”

Three-Dimensional Liver Culture Systems to Maintain Primary Hepatic Properties for Toxicological Analysis In Vitro

“…Many attempts were made to use pHHs in 3D culture with the aim to maintain and to prolong their viability and to prevent dedifferentiation. PHH spheroids were viable for at least 2 and up to 7 weeks with stable albumin production [50][51][52][53][54], urea synthesis [51,52] and glycogen storage [54]. Additionally, cellular polarization was clearly present in pHH spheroids as shown by MRP2 [51,53], P-glycoprotein (Pgp), [54], and BSEP expression [53,54].…”

“…Additionally, cellular polarization was clearly present in pHH spheroids as shown by MRP2 [51,53], P-glycoprotein (Pgp), [54], and BSEP expression [53,54]. Drug-metabolizing enzyme expression and activity of CYP1A2 [53,[55][56][57], CYP2B6 [55], CYP2C9 [55], CYP2C19 [58], CYP2D6 [53,55,57], CYP3A4 [50,53,[55][56][57][58][59], and UGTs [56,59] was stable over several weeks of pHH spheroid culture. CYP2C9 activity even increased with culture time [53,55,56,58], while CYP2C8 [53,56], and in some cases also CYP1A2, CYP2B6, CYP2D6 and CYP3A4 [55,58] activities decreased with time.…”

Exploration and application of a liver-on-a-chip device in combination with modelling and simulation for quantitative drug metabolism studies

“…This is at the lower boundary of the estimated albumin output of human liver in vivo of 37 to 105 μg per day per million cells 30 but superior to albumin production rates reported for hepatocyte spheroids recently. 31 The measurement of hepatocyte number (via hepatocyte protein amount determination in the satellite wells of each experiment) and the albumin production in all wells of an experiment allowed the hepatocyte number in each test well to be calculated and used in data processing, to account for any well to well differences in cell number (Fig. 2D).…”