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Drug metabolizing enzymes (DMEs) and transporters are important molecular determinants that affect ADME processes of xenobiotics such as drugs.For compounds that are involved with multiple mechanisms, the delineation of the relative contribution of a given mechanism, particularly the interplay between DME and transporter, is a challenging task.In the present report, a multi-tiered strategy is described, with a focus on transfected cell lines over-expressing DMEs or transporters.Mechanism-based in vivo investigations are also discussed to characterize in vitro to in vivo extrapolation.It is likely that the identification of rate-limiting steps would lead to better understanding in ADME processes and such efforts would result in safer and more effective combination therapies.