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PK In Vitro Stability (imported)

Assessment of CYP inhibition potential and DDI risk for a test compound by quantifying reversible and time-dependent (mechanism-based) inhibition across major CYP isoforms; reporting AUCR metrics and inhibition strength categories.

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Needs steward attention

Assay title "PK CIA" does not match the file name "PK In Vitro Stability". The metadata block may have been copied from another assay — owner confirmation needed. Source text contained mis-encoded characters, repaired on import.

Assay metadata

Rendered from the governed metadata model.

Assay type hierarchy
  1. bioassay
  2. ›pharmacokinetic assay
  3. ›drug–drug interaction assay
  4. ›CYP inhibition assay
  5. ›cocktail inhibition assay
Assay type *
cocktail inhibition assay
Experimental setting *
in vitro
Assay format *
cell-based
Assay design
7-plex CYP cocktail
Target *
CYP1A1CYP1A2CYP2C8CYP2C9CYP2C19CYP2D6CYP3A4
Biological process
drug metabolism
Molecule role *
test item
Cell system
human hepatocytes
Species
Not set
Donor source
Not set
Plating format
Not set
Incubation duration
Not set
Detection type *
UPLC-MS/MS
Physical detection method *
mass spectrometry
Reference gene
Not set
Expected readout *
AUCR

Description

Cocktail Inhibition Assay (CIA): The potential of test compounds to inhibit human cytochrome P450 isoforms (mainly CYP1A2, 2C8, 2C9, 2D6 and 3A4) is investigated with human hepatocytes in suspension. Test compounds (usually 5 µM) are mixed with a cocktail of CYP isoform-specific substrates (mainly phenacetin, repaglinide, diclofenac, dextromethorphan and midazolam) and incubated with hepatocytes in suspension for up to 1.5 h (co-incubation) or up to 3.5 h (pre-incubation) at 37°C to obtain clearance values. Clearance values are obtained by measurement of the substrate depletion via LC/MS analysis. As a control serves the incubation of substrate cocktail in the absence of the test compound. Inhibitory effects are determined by comparison of the clearance values for the isoform-specific substrates in the presence and absence of a potential inhibitor in co- and pre-incubation (the latter for evaluation of time-dependent inhibition). Finally, clearance ratios are calculated. A cocktail of standard inhibitors, specifically inhibiting a single CYP isoform, is always included.

Review history

This template has not been reviewed yet.

Change history

  • Imported from PK In Vitro Stability.json — flagged for steward review

    Dr Amelia Ostrowski · 18/08/2026, 19:33:36