Advancing hepatic clearance prediction across in vitro, ex situ, and in vivo systems to facilitate in vitro-to-in vivo extrapolation

Hsin-Chung Cheng; Hsueh-Tien Chen; Hsin-Ying Chen; Nai-Hsin Chou; Hong-Jaan Wang; Li-Heng Pao; Sung-Ling Tang

doi:10.1016/j.ejps.2025.107171

Advancing hepatic clearance prediction across in vitro, ex situ, and in vivo systems to facilitate in vitro-to-in vivo extrapolation

Eur J Pharm Sci. 2025 Jun 15:212:107171. doi: 10.1016/j.ejps.2025.107171. Online ahead of print.

Authors

Hsin-Chung Cheng¹, Hsueh-Tien Chen², Hsin-Ying Chen³, Nai-Hsin Chou⁴, Hong-Jaan Wang⁵, Li-Heng Pao⁶, Sung-Ling Tang⁷

Affiliations

¹ School of Pharmacy, National Defense Medical Center, Taipei, Taiwan, PR China; Graduate Institute of Pharmacy, National Defense Medical Center, Taipei, Taiwan, PR China; Medical Supplies and Maintenance, Taoyuan Armed Forces General Hospital, Taoyuan, Taiwan, PR China.
² School of Pharmacy, National Defense Medical Center, Taipei, Taiwan, PR China.
³ Graduate Institute of Life Science, National Defense Medical Center, Taipei, Taiwan, PR China.
⁴ School of Pharmacy, National Defense Medical Center, Taipei, Taiwan, PR China; Department of Pharmacy Practice, Tri-Service General Hospital, Taipei, Taiwan, PR China.
⁵ School of Pharmacy, National Defense Medical Center, Taipei, Taiwan, PR China; Graduate Institute of Pharmacy, National Defense Medical Center, Taipei, Taiwan, PR China; Graduate Institute of Life Science, National Defense Medical Center, Taipei, Taiwan, PR China.
⁶ School of Pharmacy, National Defense Medical Center, Taipei, Taiwan, PR China; Graduate Institute of Health Industry Technology, Research Center for Food and Cosmetic Safety, and Research Center for Chinese Herbal Medicine, College of Human Ecology, Chang Gung University of Science and Technology, No. 261, Wenhua 1st Road, Kweishan District, Taoyuan, PR China; Department of Gastroenterology and Hepatology, Chang Gung Memorial Hospital, Taoyuan, Taiwan, PR China.
⁷ School of Pharmacy, National Defense Medical Center, Taipei, Taiwan, PR China; Graduate Institute of Pharmacy, National Defense Medical Center, Taipei, Taiwan, PR China. Electronic address: tangling@mail.ndmctsgh.edu.tw.

PMID: 40527415
DOI: 10.1016/j.ejps.2025.107171

Abstract

Background: With rapid pharmaceutical development, hepatic clearance has become crucial for assessing drug metabolism. Rapid and effective estimation of hepatic clearance can be achieved through in vitro-to-in vivo extrapolation (IVIVE) without complicated animal experiments. However, this well-established method frequently leads to significant underestimation, limiting its clinical applicability.

Methods: We propose a series of optimizations to address these limitations. Metoprolol was selected as the target drug. In vitro microsomal assays, ex situ isolated perfused rat liver (IPRL) experiments, and in vivo pharmacokinetic studies in rats were performed to systematically investigate the factors that contribute to IVIVE deviation.

Results: To rectify the previously overlooked errors in IVIVE, we incorporated the apparent volume of distribution to refine the estimation of the intrinsic hepatic clearance derived from the Michaelis-Menten equation. Additionally, a more cytosolic-like environment was provided for microsome experiments to better simulate in vivo reactions. By integrating the findings from the IPRL experiments and pharmacokinetic studies, we identified the optimal hepatic clearance model from the perspective of liver drug metabolism-driving concentrations. Ultimately, our results indicate that the IVIVE-predicted hepatic clearance increased from the previously underestimated value of 28.1 to ∼70 mL/min/kg, which is closer to the in vivo value of 73.9 mL/min/kg. Moreover, the HEPES-KOH buffer system exhibits superior performance under these conditions.

Conclusions: We anticipate that the improved IVIVE method will provide a more comprehensive and accurate framework for predicting hepatic clearance, paving the way for its application in clinical drug dosage adjustments and new drug development.

Keywords: Hepatic clearance; In vitro to in vivo extrapolation (IVIVE); Isolated perfused rat liver (IPRL); Metoprolol; Well-stirred model (WSM).