TABLE 1

Experimental conditions for the screen assay

P450
Enzyme Amounta
Substrate
Substrate Conc.b
Positive Control
Negative Control
Excitation Wavelength
Emission Wavelength
pmol/ml μM nm
1A2 25 CEC 12 Furafylline Fluvoxamine 405 460
2C9 300 MFC 250 Tienilic acid Sulfaphenazole 405 535
3A4
50
BFC
13
Troleandomycin
Ketoconazole
405
535

  • BFC, 7-benzyloxy-4-trifluoromethylcoumarin; MFC, 7-methoxy-4-trifluoromethylcoumarin.

  • ↵ a The enzyme amount in the preincubation mix (10× required in second incubation).

  • ↵ b Substrate concentration at 4× the Km value (to reduce the effect of reversible inhibition).