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Simple Bulk Readout of Digital Nucleic Acid Quantification Assays
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Published on: September 24, 2015

A novel relay method for determining low-clearance values.

Li Di1, Patrick Trapa, R Scott Obach

  • 1Pharmacokinetics, Dynamics and Metabolism, Pfizer Inc., Eastern Point Road, Groton, CT 06340, USA. li.di@pfizer.com

Drug Metabolism and Disposition: the Biological Fate of Chemicals
|May 31, 2012
PubMed
Summary
This summary is machine-generated.

A new relay method using cryopreserved hepatocytes accurately measures low-clearance drug metabolism. This technique extends incubation times, improving predictions of in vivo clearance for drug discovery.

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Area of Science:

  • Pharmacology
  • Drug Metabolism
  • Hepatocyte Assays

Background:

  • Accurately measuring intrinsic clearance of low-clearance compounds is crucial for drug development.
  • Traditional methods using hepatocytes face challenges with prolonged incubation times and enzyme stability.

Purpose of the Study:

  • To develop and validate a novel relay method for measuring intrinsic clearance of low-clearance compounds using cryopreserved human hepatocytes.
  • To enhance the accuracy of in vitro drug metabolism assays by extending enzyme exposure time.

Main Methods:

  • A relay method involving supernatant transfer to fresh hepatocytes was employed to extend incubation.
  • Cryopreserved human hepatocytes were used to assess intrinsic clearance.
  • Seven commercial drugs with low in vivo intrinsic clearance were utilized for validation.

Main Results:

  • The relay method achieved accumulative incubation times of 20 hours or longer.
  • Predictions of human in vivo clearance using the relay method were excellent.
  • The in vitro to in vivo intrinsic clearance difference was within 2-fold for most tested compounds.

Conclusions:

  • The novel relay method is a straightforward, cost-effective, and user-friendly tool for assessing low clearance in drug discovery.
  • This method addresses key challenges in measuring drug metabolism for low-clearance compounds.
  • The relay method offers comparable prediction accuracy to standard assays for moderate to high clearance drugs.