Patent ID: 6251585
Filing Date: 2001-06-26
Classification: A61P,C12N,G01N

Abstract:
An assay for identifying an anti-mitotic agent, comprisingi. providing a culture of eukaryotic cells overexpressing a recombinant nucleic acid for a mammalian mitotic activator which activates a cyclin dependent kinase (CDK), wherein said mammalian mitotic activator is selected from the group consisting of mammalian cdc25 phosphatase and mammalian cdc2 activating kinase (CAK); and wherein said cells have an impaired cell-cycle checkpoint caused by decreased inhibitory phosphorylation of the CDK or increased activating phosphorylation of the CDK which causes premature entry of the cells into mitosis so as to cause cell death, the premature entry into mitosis being mediated at least in part by the activation of the CDK by the mitotic activator;ii. contacting the culture of cells with a candidate agent under conditions wherein the cell-cycle checkpoint is impaired;iii. measuring a level of proliferation of the cells in the presence of the candidate agent; andiv. comparing the level of proliferation of the cells in the presence of the candidate agent to a level of proliferation of the cells in the absence of the candidate agent, wherein an increase in the level of proliferation in the presence of the candidate agent is indicative of anti-mitotic activity of the candidate agent.