TABLE 1

Enzyme kinetic parameters for the formation of M7 in incubations of brivanib with HLM, cDNA-expressed CYP3A4, and cDNA-expressed CYP1A2

Incubations were conducted at 37°C with HLM (0.1 mg protein/ml), CYP3A4 (25 pmol/ml), or CYP1A2 (10 pmol/ml) in the presence of NADPH. Enzyme kinetic parameters were estimated by fitting the data to the Michaelis-Menten equation using a nonlinear regression analysis with GraphPad Prism (version 5.04; GraphPad Software Inc.). The corresponding r² values for the curves ranged from 0.90 to 0.94.

Enzyme	K_m	V_max^{^a}	CL_int^{^b}
	μM		V_max/K_m
HLM	29.1 ± 7.9	89.1 ± 9.8	3.1
CYP3A4	32.7 ± 10.9	0.53 ± 0.11	0.016
CYP1A2	45.0 ± 16.2	1.39 ± 0.25	0.031

↵a The unit for V_max was picomole per milligram of protein per minute for HLM and picomole per picomole of enzyme per minute for expressed enzymes.
↵b The unit for CL_int was microliter per milligram of protein per minute for HLM and microliter per picomole of enzyme per minute for expressed enzymes.