Comparison of intrinsic clearances in human liver microsomes and suspended hepatocytes from the same donor livers: clearance-dependent relationship and implications for prediction of in vivo clearance

Abstract

1. Intrinsic clearance (CL_int) of seven probe cytochrome P450 substrates, across a wide range of clearance, was compared in microsomes and cryopreserved hepatocytes from the same four livers. Previous comparisons have shown system dependence, but using preparations from different donor livers.

2. Four-fold average underprediction of microsomal CL_int by hepatocytes (scaled to whole liver) for high clearance substrates (midazolam, nifedipine, and diclofenac) was observed with relatively unbiased prediction (within 1.5-fold average) for the low/medium clearance substrates (tolbutamide, alprazolam, bufuralol, and triazolam).

3. CL_int of midazolam and nifedipine corresponded between livers over a tenfold range, but the absolute ranges were lower for hepatocytes, indicating independence of hepatocyte bias from substrate. In contrast, the absolute ranges of CL_int for the low clearance CYP3A4 substrate alprazolam were similar between the systems, indicating independence of hepatocyte bias from enzyme.

4. The trend in CL_int between the systems was similar to that in a dataset of published CL_int for 46 substrates in microsomes and hepatocytes (unrelated liver sources), supporting a fundamental rate limitation of the hepatocyte system.

5. A tendency of decreasing V_max in hepatocytes relative to microsomes, with increasing clearance, suggests that a capacity limitation, such as cofactor rate limitation, may be involved in this phenomenon.

Keywords::

• In vitro
• cytochrome P450
• CYP3A4
• CYP2C9
• CYP2D6