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Atypical Cytochrome P450 Kinetics

Implications for Drug Discovery

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Abstract

The Michaelis-Menten model is commonly used to estimate a drug’s potential in vivo hepatic clearance based on in vitro data obtained during drug discovery and development. This paradigm assumes that the drug obeys ‘typical’ enzyme kinetics and thus can be described by this model. However, it is increasingly being recognised that a number of drugs metabolised not only by the cytochrome P450 enzymes but also by other enzymes and transporters can exhibit atypical kinetic profiles, and thus are not accurately modeled with the Michaelis-Menten model. Application of an incorrect model can then lead to mis-estimation of in vitro intrinsic clearance and thus affect the prediction of in vivo clearance. This review discusses several types of atypical kinetic profiles that may be observed, including examples of homotropic cooperativity (i.e. sigmoidal kinetics, biphasic kinetics and substrate inhibition kinetics) as well as heterotropic cooperativity (i.e. activation). Application of the incorrect kinetic model may profoundly affect estimations of intrinsic clearance. For example, incorrectly applying the Michaelis-Menten model to a kinetic profile exhibiting substrate inhibition kinetics will result in an underestimation of Km (Michaelis-Menten constant) and Vmax (maximal velocity), whereas application of the Michaelis-Menten model to sigmoidal kinetic data typically results in an overestimation of Km and Vmax at the lower substrate concentrations that are typically therapeutically relevant. One must also be careful of potential artefactual causes of atypical kinetic profiles, such as enzyme activation by solvents, buffer dependent kinetic profiles, or altered kinetic parameter estimates due to nonspecific binding of the substrate to proteins. Despite a plethora of data on the effects of atypical kinetic profiles in vitro, only modest effects have been noted in vivo (with the exception of substrate dependent inhibition). Thus, the clinical relevance of these phenomena remains inconclusive.

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Acknowledgements

Supported in part by Public Health Service grant #GM 063215 from the National Institutes of Health.

The author has no conflicts of interest that are directly relevant to the content of this review.

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  1. Department of Experimental and Clinical Pharmacology, College of Pharmacy, University of Minnesota, 308 Harvard St SE, Minneapolis, MN, 55455, USA

    Timothy S. Tracy

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Tracy, T.S. Atypical Cytochrome P450 Kinetics. Drugs R D 7, 349–363 (2006). https://doi.org/10.2165/00126839-200607060-00004

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