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Impact of time-dependent inactivation on the estimation of enzyme kinetic parameters for midazolam.

Enzyme kinetic parameters for midazolam were time-dependent in human liver microsomes, under initial velocity conditions. V(max) and K(m) decreased up to 3.7 and 3.1-fold, respectively, for 10 min compared to 1 min incubations. Mathematical models describing the relationship between inactivation and the time-dependency of enzyme kinetic parameter estimates were derived and discussed.