IDENTIFICATION OF A SERIES OF POTENT TELOMERASE INHIBITORS USING A TIME-RESOLVED FLUORESCENCE-BASED ASSAY

A high-throughput, nonradioactive telomerase assay using a europium-la beled oligonucleotide probe and time-resolved fluorescence has been de veloped to detect PCR-amplified telomerase products. The use of a ther mally activated TAQ polymerase, rather than a wax barrier, to implemen t a PCR hot-start facilitated the use of laboratory automation equipme nt. Results obtained with this high-throughput protocol correlate well with those obtained with the TRAP protocol. Screening a set of 125,00 0 compounds from the Berlex library, we identified a set of isothiazol one-containing telomerase inhibitors. The most potent of these inhibit ors have submicromolar IC50 values and may be reacting with a telomera se thiol. These compounds may be useful tools to evaluate the effects of telomerase in cancer cells. (C) 1998 Wiley-Liss, Inc.