Large-scale chemical-genetics yields new M. tuberculosis inhibitor classes
Large-scale chemical-genetics yields new M. tuberculosis inhibitor classes
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DOI：
10.1038/s41586-019-1315-z
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发表时间：
2019-07-04
期刊：
影响因子：
64.8
通讯作者：
Hung, Deborah T.
中科院分区：
文献类型：
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作者：
Johnson, Eachan O.;LaVerriere, Emily;Hung, Deborah T.
需要新的抗生素来对抗不断上升的耐药性，其中新的结核分枝杆菌 (Mtb) 药物具有最高优先级。然而，传统的全细胞和生化抗生素筛选已经失败。在这里，我们制定了一种名为 PROSPECT（对菌株进行初步筛选，以优先考虑扩展化学和靶标）的策略，其中我们针对缺乏基本细菌靶标的菌株池筛选化合物。我们设计了针对 474 个 Mtb 必需基因的菌株，并针对活性丰富且无偏见的化合物库筛选了 100-150 个菌株库，探测了超过 850 万个化学-遗传相互作用。初级筛选发现的命中率比单独筛选野生型结核分枝杆菌多十倍以上，化学-遗传相互作用提供了即时、直接的目标洞察。我们鉴定了超过 40 种针对 DNA 旋转酶、细胞壁、色氨酸、叶酸生物合成和 RNA 聚合酶的化合物，以及针对 EfpA 的抑制剂。化学优化产生了具有有效野生型活性的 EfpA 抑制剂，从而证明了 PROSPECT 能够针对传统药物发现无法发现的靶点产生抑制剂。
New antibiotics are needed to combat rising levels of resistance, with new Mycobacterium tuberculosis (Mtb) drugs having the highest priority. However, conventional whole-cell and biochemical antibiotic screens have failed. Here we develop a strategy termed PROSPECT (primary screening of strains to prioritize expanded chemistry and targets), in which we screen compounds against pools of strains depleted of essential bacterial targets. We engineered strains that target 474 essential Mtb genes and screened pools of 100-150 strains against activity-enriched and unbiased compound libraries, probing more than 8.5 million chemical-genetic interactions. Primary screens identified over tenfold more hits than screening wild-type Mtb alone, with chemical-genetic interactions providing immediate, direct target insights. We identified over 40 compounds that target DNA gyrase, the cell wall, tryptophan, folate biosynthesis and RNA polymerase, as well as inhibitors that target EfpA. Chemical optimization yielded EfpA inhibitors with potent wild-type activity, thus demonstrating the ability of PROSPECT to yield inhibitors against targets that would have eluded conventional drug discovery.