Z-338 - Cerep

Method: In vitro metabolism study
HPLC methods that were validated for the quantification of Z-338,
Z-338 deisopropyl metabolite and ID951551 were employed.
Z-338 and ID951551 were incubated with a panel of recombinant
human enzymes to investigate possible involvement of CYP and
UGT enzymes responsible for the metabolism of Z-338 and
ID951551. rCYP and rUGT enzymes were obtained from Gentest
Corporation (Woburn, MA) or Cypex Ltd, (Scotland, UK) . Once
initial rate conditions were established, Km and Vmax were
determined for the dealkylation of Z-338 by human liver
microsomes (HLM), recombinant CYP2C8 and recombinant
CYP3A4. HLM was prepared by XenoTech (Lenexa, KS).
An antibody inhibition experiment was performed to determine the
relative contribution of CYP2C8 and CYP3A4 to the metabolism of
Z-338 and ID951551 by HLM. Polyclonal antibodies and intact
rabbit serum were obtained from Nosan Corporation (Yokohama,
Japan).