Patent ID: 7968314

Claim:
A method of evaluating whether a test compound is a slow or a rapid acting inhibitor of a target enzyme so as to select time points for a time-dependent inhibition assay, comprising the steps of: a. performing a first assay of the test compound, said first assay comprising the steps of: i. preparing a first set of samples of the test compound and the target enzyme, said first set including samples containing the target enzyme and varying concentrations of the test compound; ii. preparing a second set of samples of the test compound and the target enzyme, said second set including samples containing the target enzyme and varying concentrations of the test compound; iii. evaluating the amount of products formed from reactants in said first set of samples after diluting the samples at a first time interval to obtain first results; and iv. evaluating the amount of products formed from reactants in said second set of samples after diluting the samples at a second time interval to obtain second results, said second time interval being different from said first time interval; b. comparing said first and second results to determine whether the test compound is a slow acting inhibitor that substantially inhibits the target enzyme in more than about 10 minutes or a rapid acting inhibitor that inhibits the target enzyme in about 10 minutes or less; c. selecting time points for a secondary inhibition assay of the test compound to determine time-dependent kinetics based on whether the test compound is a slow acting inhibitor or a rapid acting inhibitor; and d. performing the secondary inhibition assay, said secondary inhibition assay comprising the steps of: i. preparing a secondary set of samples of the test compound and the target enzyme, said secondary set including samples containing the target enzyme and varying concentrations of the test compound; and ii. evaluating the amount of products formed from reactants in said secondary set of samples after diluting the samples at said selected time points.