Kinetic analysis
Rivaroxaban (1−100 μM) was incubated with pooled HLM
(0.3
mg protein·mL−1) for 60 min to evaluate the kinetic
parameters of the HLM. Rivaroxaban (1−200 μM for CYP2J2, 10−200 μM for
CYP 2D6 and 20−200 μM for CYP4F3) was incubated with recombinant CYPs
(0.4 mg protein·mL−1) for 60 min to determine the
kinetic parameters of the recombinant CYPs. The incubation conditions
were as described above. The apparent K m andV max values were calculated from nonlinear
regression analysis of experimental data according to the
Michaelis-Menten equation with GraphPad Prism, version 7. All
incubations were carried out in three independent experiments performed
in duplicate.