Table 2. Effects of Core Modifications on TDI.

Test compound (50 μM with serial dilutions) is incubated with human liver microsomes (0.5 mg/mL) in phosphate buffer (100 mM) with NADPH (1 mM) for 0, 5, 15, and 30 min. The incubation mixture is diluted 20-fold then incubated with CYP3A4 substrate midazolam (20 μM) to determine residual CYP3A4 enzyme activity. The enzyme activity vs incubation time is plotted to obtain kinetic parameters for TDI.