In vivo pharmacokinetics and pharmacodynamics study
Non-fasting C57BL/6J mice were orally administered CDDO-Me at doses of 3
and 10 mg・kg-1. Blood samples were collected via the
abdominal vein into blood collection tubes containing EDTA-2K (Terumo
Corp, Tokyo, Japan) under anaesthesia at 1, 4, 6, and 24 h after oral
administration. Liver samples were also collected to analyse the
expression of Nqo1 using quantitative PCR. The plasma
concentration of CDDO-Me was measured using the API 3200 triple
quadrupole mass spectrometer (AB Sciex LLC., Redwood City, CA) under the
following conditions: column, Kinetex C18 (Phenomenex Inc., Torrance,
CA, 50 × 2.1 mm, 2.6 μm); column temperature, 40 °C; flow rate, 0.4
mL・min-1; mobile phase A, 0.1% formic acid/MilliQ
aqueous solution; mobile phase B, 0.1% formic acid in 50
v・v-1% acetonitrile/methanol solution; and gradient
– 0 min: A / B=90 / 10, 1 min: A / B=10 / 90, 1-3 min: A / B=10 / 90,
3.01 min, A / B: 90 / 10. CDDO-Me was quantified in the negative
ionization mode using multiple reaction monitoring with niflumic acid as
the internal standard, as previously described (Resham et al., 2015).
Data were acquired and integrated using Analyst software version 1.7 (AB
Sciex LLC., RRID: not yet assigned).