Metabolic stability assay in hepatocytes

Aim: Screening of in vitro metabolic stability of test compound in cryopreserved human and preclinical species hepatocytes.

Background information: Metabolism is the enzymatic modification of drugs to enable their excretion. Although metabolites are typically inactive, some metabolites may be pharmacologically active.

The liver is the main site of drug metabolism in the body. Unlike liver microsomes, hepatocytes contain both phase I and phase II compound metabolising enzymes, which are present in the intact cell.

Metabolic stability is assessed by measuring test compound depletion with time. Results can be used to assess interspecies differences in metabolism and in vitro intrinsic clearance, which in turn, can be used to predict in vivo hepatic clearance.

Compound requirement: 50 µl of 10 mM DMSO solution

Turnaround time: 4 weeks / 2 compounds / 5 species

Experimental design:

  • Test compound concentration: 1 µM (variable on request)
  • Number of replicates: 2
  • Number of cells: 1 million/ml
  • Time points: 0, 30, 60, 90 and 120 min ± cells, and negative control
  • Positive control: compound with known stability
  • Analytical method: UPLC-MS/MS

Delivered results:

  • In vitro intrinsic clearance
  • % test compound remaining at each time point relative to time 0