Drug metabolism in dog liver microsomes assessed as ER-879123 formation at 10 uM after 60 mins in presence of NADH and carboxylesterase inhibitor BNPP by LC-MS/MS method
Drug metabolism in human liver microsomes assessed as ER-879123 formation at 10 uM after 60 mins in presence of NADH and carboxylesterase inhibitor BNPP by LC-MS/MS method
Drug metabolism in dog liver microsomes assessed as ER-879123 formation at 10 uM after 60 mins in presence of NADH and FMO1 and FMO3 substrate/inhibitor methimazole by LC-MS/MS method
Drug metabolism in human liver microsomes assessed as ER-879123 formation at 10 uM after 60 mins in presence of NADH and FMO1 and FMO3 substrate/inhibitor methimazole by LC-MS/MS method
PARP Enzyme Assay: The PARP enzyme assay is set up on ice in a volume of 100 microliters consisting of 100 mM Tris-HCl (pH 8.0), 1 mM MgCl2, 28 mM KCl, 28 mM NaCl, 0.1 mg/ml of DNase I activated herring sperm DNA (Sigma, Mo.), 3.0 micromolar [3H]nicotinamide adenine dinucleotide (470 mci/mmole), 7 micrograms/ml PARP enzyme, and various concentrations of the compounds to be tested. The reaction is initiated by incubating the mixture at 25° C. After 15 minutes of incubation, the reaction is terminated by adding 500 microliters of ice cold 20% (w/v) trichloroacetic acid. The precipitate formed is transferred onto a glass fiber filter (Packard Unifilter-GF/B) and washed three times with ethanol. After the filter is dried, the radioactivity is determined by scintillation counting.