A High-Throughput Assay for Developing Inhibitors of PhoP, A Virulence Factor of Mycobacterium tuberculosis.

A High-Throughput Assay for Developing Inhibitors of PhoP, A Virulence Factor of Mycobacterium tuberculosis.
Comb Chem High Throughput Screen. 2016 Oct 10;
Authors: Wang L, Xu M, Southall N, Zheng W, Wang S
Abstract
Tuberculosis (TB) still kills over 1.5 million people per year despite available anti-TB drugs. The emergence of drug-resistant TB poses a major threat to public health and prompts for an urgent need for new and more effective drugs. The long duration needed to treat TB by the current TB drugs, which target the essential cellular activities, inevitably leads to the emergence of drug-resistance. PhoP of Mycobacterium tuberculosis (MTB), an essential virulence factor, is a novel target for drug development. Guided by the crystal structure of the PhoP-DNA complex, we designed and developed a robust high-throughput screening assay for identification of PhoP inhibitors that disrupt the PhoP-DNA binding. The assay was in Foster resonance energy transfer (FRET) format with a Cy3 dye linking to DNA and Cy5 to PhoP. Three active compounds were identified that inhibited PhoP-DNA interactions by binding directly to PhoP after a test screening of ~6,000 bioactive compounds and approved drugs. While these three PhoP inhibitors may be useful for further development, our results demonstrate that this FRET based PhoP-DNA binding assay is valid for additional compound library screening to identify new leads for developing novel TB drugs t...

Source: Combinatorial Chemistry and High Throughput Screening - October 9, 2016 Category: Chemistry Authors: Wang L, Xu M, Southall N, Zheng W, Wang S Tags: Comb Chem High Throughput Screen Source Type: research