Studying cytochrome P450 kinetics in drug metabolism

Abstract

Background: Determination of cytochrome P450 enzyme-mediated kinetics in vitro can be useful for predicting drug dosing and clearance in humans. Expressed P450s, human liver microsomes, human hepatocytes (both fresh and cryopreserved), and human liver slices are used to estimate K_m and V_max values for determination of intrinsic clearance of the drug for scale-up to predict in vivo clearance. Objective: To describe the advantages and disadvantages of the various in vitro systems used to estimate kinetic parameters for disposition of drugs and the various kinetic profiles that can be observed. Methods: A review of the literature was conducted to evaluate the utility of the various in vitro preparations, the methods for determining kinetic parameters and the types of kinetic profiles that may be observed. Results/conclusions: The choice of in vitro system for determining kinetic parameters will depend on the objective of the studies, as each system has advantages and disadvantages. Kinetic parameter determinations must be carefully assessed to assure that the correct kinetic model is applied and the most accurate kinetic parameters are determined.

Keywords::

• atypical
• biphasic
• cytochrome P450
• hepatocytes
• heterotropic activation kinetics
• K_m
• microsomes
• sigmoidal
• substrate inhibition
• V_max

Acknowledgement

Supported in part by NIH grant #GM063215 and #GM069753.