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Determination of Fatty Acid Oxidation and Lipogenesis in Mouse Primary Hepatocytes
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An Integrated Hepatocyte Stability Assay for Simultaneous Metabolic Stability Assessment and Metabolite Profiling.

Christian Leung1, Joyce Liu1, Katherine Cunico1

  • 1Department of Drug Metabolism and Pharmacokinetics, Genentech, Inc., South San Francisco, California.

Drug Metabolism and Disposition: the Biological Fate of Chemicals
|March 4, 2024
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Summary

A new integrated assay streamlines drug discovery by simultaneously assessing metabolic stability and metabolite profiles in hepatocytes. This high-throughput method accelerates lead optimization and reduces resource demands.

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Area of Science:

  • Drug Discovery and Development
  • Pharmacokinetics and Metabolism
  • Analytical Chemistry

Background:

  • Metabolic stability and metabolite identification are crucial for optimizing drug candidates and prioritizing compounds in early drug discovery.
  • Current in vitro assays for metabolic stability and metabolite profiling are resource-intensive, requiring extensive sample preparation and time.
  • Understanding metabolic liabilities guides medicinal chemists in the structural design of new chemical entities.

Purpose of the Study:

  • To develop a high-throughput integrated assay for simultaneous hepatocyte metabolic stability assessment and metabolite profiling.
  • To overcome the time and resource limitations associated with traditional, separate metabolic assays.
  • To accelerate the lead optimization process in drug discovery through an efficient and combined workflow.

Main Methods:

  • An automated liquid-handling system for sample preparation and incubation.
  • A liquid chromatography-high-resolution mass spectrometry system for simultaneous monitoring of parent compound depletion and metabolite formation.
  • Automated data analysis software for hepatic clearance assessment and streamlined autobatch processing for metabolite profiling.

Main Results:

  • The integrated assay platform was validated using eight control compounds across three species, demonstrating accuracy, repeatability, and broad metabolite coverage.
  • Simultaneous assessment of metabolic stability and metabolite profiles was achieved.
  • The developed platform significantly improved the efficiency of demanding screening assays.

Conclusions:

  • The integrated assay platform, combining automation, high-resolution mass spectrometry, and batch-processing software, enhances the efficiency of metabolic stability and metabolite profiling.
  • This approach allows for simultaneous determination of metabolic stability and metabolite profiles, accelerating lead optimization in a resource-effective manner.
  • The developed assay is pivotal for comprehending metabolic liabilities and optimizing chemical entities in drug discovery programs.