TABLE 5

Effects of microsomal protein concentration on the unbound (free) concentration of selected metabolism-dependent inhibitors and the extent of inhibitor depletion

All values were rounded to two significant figures.

Inhibitor and [HLM]	NADPH	Inhibitor Loss
Inhibitor and [HLM]	NADPH	HLM Binding^{^a}	Metabolism after 30 min^{^b}	Metabolism + HLM Binding^{^c}
		%
Ticlopidine (0.2 μM)
0.1 mg/ml	−	30		30
1.0 mg/ml	−	81		81
0.1 mg/ml	+		98	100
1.0 mg/ml	+		100	100
Tienilic acid (0.5 μM)
0.1 mg/ml	−	0		0
1.0 mg/ml	−	12		12
0.1 mg/ml	+		28	28
1.0 mg/ml	+		100	100
S-Fluoxetine (10 μM)
0.1 mg/ml	−	46		46
1.0 mg/ml	−	78		78
0.1 mg/ml	+		21	52
1.0 mg/ml	+		21	91
Paroxetine (0.2 μM)
0.1 mg/ml	−	55		55
1.0 mg/ml	−	86		86
0.1 mg/ml	+		53	74
1.0 mg/ml	+		88	98
Azamulin (0.1 μM)
0.05 mg/ml	−	2.5		2.5
0.5 mg/ml	−	24		24
0.05 mg/ml	+		11	11
0.5 mg/ml	+		99	100
Mibefradil (0.2 μM)
0.05 mg/ml	−	64		64
0.5 mg/ml	−	83		83
0.05 mg/ml	+		85	97
0.5 mg/ml	+		100	100

↵a HLM binding was attributed to the observed loss of inhibitor in the absence of NADPH in the postcentrifugation supernatant fraction.
↵b The observed loss of inhibitor due to metabolism alone was determined after a 30-min incubation with HLM in the presence of NADPH followed by quenching the entire reaction with acetonitrile and assessment of remaining inhibitor.
↵c Contribution of metabolism and HLM binding to depletion of inhibitor. Total inhibitor loss determined after a 30-min incubation with HLM in the presence of NADPH followed by centrifugation and analysis of remaining inhibitor present in the supernatant fraction.