Infectious Disease Research Institute, Seattle, Washington, United States of America.
PLoS One. 2019 Jan 16;14(1):e0205479. doi: 10.1371/journal.pone.0205479. eCollection 2019.
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.
结核病是一种具有全球重要性的疾病，迫切需要新型药物。我们开发了一种全细胞表型筛选方法，可用于鉴定抑制分枝杆菌生长的抑制剂。我们使用表达远红荧光报告基因的毒力重组分枝杆菌菌株，并使用荧光监测体外生长。我们使用 96 孔和 384 孔板优化了高通量测定法；这两种格式都提供了符合严格重现性和稳健性测试的测定法。我们筛选了先前显示对分枝杆菌有效的 1105 种化学多样性化合物的化合物组，并确定了显示≥90％生长抑制的主要命中。我们对命中进行了排名，并确定了三类感兴趣的化合物：苯氧烷基苯甲酰胺，苯并噻吩 1-1 二恶烷和哌啶胺。这些新的化合物类可能可作为开发防止分枝杆菌生长的新型抑制剂系列的起点。该测定法可用于进一步筛选，也可以轻松适应分枝杆菌的其他菌株。
