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Related Experiment Video

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Assessment of Bile Salt Export Pump (BSEP) Inhibition in Membrane Vesicles Using Radioactive and LC/MS-Based

Lisa D Marroquin1, Paul D Bonin2, Julie Keefer1

  • 1Hit Discovery and Optimization, Pfizer Inc, Groton, Connecticut.

Current Protocols in Toxicology
|February 2, 2017
PubMed
Summary

New assays detect drug interference with the bile salt export pump (BSEP). This helps identify potential liver injury risks early, preventing drug withdrawal and liver failure.

Keywords:
ATP-binding cassette transporterBSEPDILIbile salt export pumpcholestasisdrug induced liver injury

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Area of Science:

  • Hepatology
  • Drug Metabolism
  • Molecular Biology

Background:

  • The bile salt export pump (BSEP, ABCB11) is crucial for bile salt secretion in the liver.
  • BSEP inhibition by drugs can cause bile salt accumulation, leading to cholestasis and drug-induced liver injury (DILI).
  • DILI is a significant factor in drug market withdrawals and acute liver failure.

Purpose of the Study:

  • To develop and describe assays for detecting drug-induced BSEP inhibition.
  • To investigate the link between drug interference with BSEP function and liver injury.
  • To provide protocols for early identification of BSEP-associated DILI risks.

Main Methods:

  • Utilized membrane vesicles from Hi5 insect cells expressing human BSEP.
  • Developed assays in a 384-well format for high-throughput screening.
  • Employed both radiolabeled and liquid chromatography/mass spectrometry (LC/MS) detection methods.

Main Results:

  • Established comprehensive protocols for assessing BSEP inhibition.
  • Demonstrated methods for quantifying drug effects on BSEP transporter activity.
  • Provided a framework for evaluating potential DILI associated with BSEP inhibition.

Conclusions:

  • The described assays enable efficient detection of BSEP inhibition by drug candidates.
  • Early identification of BSEP inhibitors can mitigate the risk of DILI.
  • These methods support safer drug development by assessing BSEP-associated liver injury potential.