A novel relay method for determining low-clearance values.


A novel relay method has been developed using cryopreserved human hepatocytes to measure intrinsic clearance of low-clearance compounds. The relay method involved transferring the supernatant from hepatocyte incubations to freshly thawed hepatocytes at the end of the 4-h incubation to prolong the exposure time to active enzymes in hepatocytes. An accumulative incubation time of 20 h or longer in hepatoctyes can be achieved using the method. The relay method was validated using seven commercial drugs (diazepam, disopyramide, theophylline, timolol, tolbutamide, S-warfarin, and zolmitriptan) that were metabolized by various cytochrome P450s with low human in vivo intrinsic clearance at approximately 2 to 15 ml · min⁻¹ · kg⁻¹. The results showed that the relay method produced excellent predictions of human in vivo clearance. The difference between in vitro and in vivo intrinsic clearance was within 2-fold for most compounds, which is similar to the standard prediction accuracy for moderate to high clearance compounds using hepatocytes. The relay method is a straightforward, relatively low cost, and easy-to-use new tool to address the challenges of low clearance in drug discovery and development.

DOI: 10.1124/dmd.112.046425

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@article{Di2012ANR, title={A novel relay method for determining low-clearance values.}, author={Li Di and Patrick E. Trapa and Ronald Scott Obach and Karen A Atkinson and Yi-an Bi and Angela C. Wolford and Beijing Tan and Thomas S. McDonald and Yurong Lai and Larry M. Tremaine}, journal={Drug metabolism and disposition: the biological fate of chemicals}, year={2012}, volume={40 9}, pages={1860-5} }