Abstract
Tuberculosis is a disease of global importance for which novel drugs are urgently required. We developed a whole-cell phenotypic screen which can be used to identify inhibitors of Mycobacterium tuberculosis growth. We used recombinant strains of virulent M. tuberculosis which express far-red fluorescent reporters and used fluorescence to monitor growth in vitro. We optimized our high throughput assays using both 96-well and 384-well plates; both formats gave assays which met stringent reproducibility and robustness tests. We screened a compound set of 1105 chemically diverse compounds previously shown to be active against M. tuberculosis and identified primary hits which showed ≥ 90% growth inhibition. We ranked hits and identified three chemical classes of interest–the phenoxyalkylbenzamidazoles, the benzothiophene 1–1 dioxides, and the piperidinamines. These new compound classes may serve as starting points for the development of new series of inhibitors that prevent the growth of M. tuberculosis. This assay can be used for further screening, or could easily be adapted to other strains of M. tuberculosis.
Highlights
Tuberculosis (TB), caused by the bacterial pathogen Mycobacterium tuberculosis, is a disease of global importance which killed approximately 1.7 million people in 2016 [1]
High throughput screening of small molecules has the potential to identify new compound classes that are effective against M. tuberculosis
We were interested in developing a simple whole cell screen which could be used in multiple formats to assess the anti-tubercular activity of large compound sets
Summary
Tuberculosis (TB), caused by the bacterial pathogen Mycobacterium tuberculosis, is a disease of global importance which killed approximately 1.7 million people in 2016 [1]. A lengthy 4-drug regimen is used to treat active infection, but drug resistant strains have emerged and threaten efforts to control the disease. Multi-drug resistant (MDR) and extremely drug resistant (XDR) TB are gaining footholds in areas where HIV is predominant and/or antibiotic treatment of patients is administered incompletely or incorrectly [1]. There is an urgent need for new drugs that are effective at killing M. tuberculosis and which might shorten therapy. High throughput screening of small molecules has the potential to identify new compound classes that are effective against M. tuberculosis. A number of targets have been tested [2,3,4,5,6,7,8,9,10,11,12,13,14,15] but this approach had limited success in finding hits with whole cell activity for a variety of reasons including lack of permeation, efflux, and poor target vulnerability or engagement [16]
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