CYP3A induction by N-hydroxyformamide tumor necrosis factor-α converting enzyme/matrix metalloproteinase inhibitors: Use of a pregnane X receptor activation assay and primary hepatocyte culture for assessing induction potential in humans

A series of N-hydroxyformamide tumor necrosis factor-α converting enzyme (TACE)/matrix metalloprotease (MMP) inhibitors were evaluated for their potential to induce human cytochrome P450 3A (CYP3A). Two in vitro assays were used: 1) a cell-based reporter gene assay for activation of the pregnane X receptor (PXR), and 2) a primary "sandwich" culture of human hepatocytes. Approximately 50 TACE/MMP inhibitors were evaluated in the human PXR assay. A range of PXR activation was observed, 0 to 150% of the activation of the known human CYP3A inducer rifampicin. Three TACE/MMP inhibitors were evaluated in rat and human hepatocytes. Significantly higher PXR activation/CYP3A induction was observed in PXR/hepatocyte models, respectively, for (2R,3S) 3-(formyl-hydroxyamino)-2-(2-methyl-1-propyl)-4-methylpentanoic acid [(1S,2S)-2-methyl-l-(2- pyridylcarbamoyl)-l-butyl]amide (GW3333) compared with (2R,3S)6,6,6-trifluoro-3-[formyl(hydroxy)amino]-2-isobutyl-N-{(1S,2R)-2- methoxy-1-[(1,3-thiazol-2-ylamino)carbonyl]propyl}hexanamide (GW6495) and (2R)-N-{(1S)-2,2-dimethyl-1-[(methylamino)carbonyl]-propyl}-2-{(1S)-1- [formyl(hydroxy)amino]ethyl}-5 -phenylpentanamide (GI4023). The CYP3A induction level achieved with GW3333 at a concentration of approximately 10 μM in human hepatocytes was comparable to that achieved with rifampicin at a concentration of 10 μM. The extent of rodent CYP3A induction caused by GW3333 was confirmed in vivo after daily oral administration for 14 days to rats. In conclusion, GW3333 is a potential inducer of CYP3A expression in vivo in humans, but other N-hydroxyformamides are less likely to induce CYP3A.