Inhibitor and [HLM] | NADPH | Inhibitor Loss | ||
---|---|---|---|---|
HLM Bindinga | Metabolism after 30 minb | Metabolism + HLM Bindingc | ||
% | ||||
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.