METHODS FOR IDENTIFYING NOTCH-SPARING GAMMA SECRETASE INHIBITORS

摘要

The invention herein provides a method for identifying a Notch-sparing ?-secretase inhibitor (NS-GSI) by evaluating e- and ?-cleavage in a single APP-based substrate. The inventive assay utilizes e-cleavage as surrogate for S3-cleavage of Notch and a substrate comprising the C99 fragment of APP fused to a Gal4/VP16 transcription factor to evaluate the inhibitory activity of compounds on e and ?-cleavage simultaneously and for the identification of e -sparing inhibitors that decrease Aß production without affecting AICD production. In another embodiment, the APP-based substrate incorporates a mutated a-secretase cleavage site for enhanced Aß production which allows for greater sensitivity of the assay.