In vitro-in vivo extrapolation of intrinsic clearance based on data from human hepatocyte spheroids – a case study with the low-clearance drug xevinapant

Abstract Xevinapant, a drug previously under development for head and neck cancer, was investigated to determine its intrinsic clearance (CLint) in vitro and assess the involvement of CYP enzymes in its metabolism. Extensive in vitro studies showed very low turnover, necessitating advanced methodologies to accurately measure CLint values. Two novel in vitro assays, using a modified suspension hepatocyte protocol or 3D hepatocyte spheroid cultures, allowed measuring hepatic turnover of xevinapant with very low CLint values of 1.3 or 0.3 µL/min/106 cells, respectively. Reaction phenotyping using specific inhibitors, enabled by enhanced detection capabilities in spheroids, identified primarily CYP3A (> 50%) as contributor to the hepatic metabolism of xevinapant. In vitro-in vivo extrapolation, in combination with data from the human mass balance study, revealed that hepatic metabolism, primarily mediated by CYP3A, accounts for approximately 30% of the overall clearance. The remaining clearance is renal (∼20%) and metabolic, but potentially extrahepatic (∼50%). This research highlights the importance and benefits of utilising advanced in vitro techniques, such as human hepatocyte spheroids, to accurately characterise drug metabolism, providing valuable insights for pharmacokinetic evaluations and, ultimately, supporting the development of a PBPK model and understanding the DDI risk of xevinapant.

• Publication year

  2025

• Venue

  Xenobiotica; the fate of foreign compounds in biological systems

• Publication date

  2025-11-10

• Fields of study

  Medicine

• Identifiers
  DOI 10.1080/00498254.2025.2587283PMID 41211641
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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