Patent ID: 6489094
Filing Date: 2002-12-03
Classification: B01L,C12Q,G01N

Abstract:
A method for preparing inhibitor compounds for in vitro testing of in vivo drug-drug interactions in mammals, wherein the method comprises the automated steps of:a) retrieving frozen microsomes of a mammal species from a freezer storage unit; b) thawing the retrieved frozen microsomes; c) mechanically introducing a pre-determined quantity of (i) the thawed microsomes, (ii) a cofactor, and (iii) a buffer into individual wells of a multiwell reaction plate and pre-incubating the reaction plate with contained microsomes to an in vivo temperature for said mammal for a pre-determined period of time; d) mechanically combining the thawed microsomes in each of the wells of the reaction plate with a pre-determined quantity of a combination of one of said inhibitor compounds and a preselected probe substrate specific for cytochrome P450s (CYP); e) incubating the reaction plate with contained microsomes, CYP-specific probe substrate and inhibitor compounds to said in vivo temperature for a pre-determined period of time, to effect reaction in the individual wells, with resultant respective reaction products; f) stopping any reactions which occur, at the end of the pre-determined period of time, purifying the reaction products and using the reaction products in an analysis test for determining the extent of any drug-drug interactions relative to the respective inhibitor compounds and respective CYP-specific probe substrate; wherein steps c-f are repeated for a pre-determined number of times as individual test runs, with pre-thawed microsome samples, wherein, simultaneously with the steps of c-f of one run, additional microsomes are removed from the freezer unit, thawed, and held at a holding site, for immediate use in steps c-f of a subsequent test run, wherein timing of when the frozen microsomes are initially removed for thawing during steps c-f is such that removal is at the same point in time during said steps of each of the individual test runs and wherein duration of thawing and any pre-incubation is the same for all of the runs and incubation periods are pre-calculated and adapted to be of the same duration for each paired set of plates during a test run.