Identification of novel inhibitors of M. tuberculosis growth using whole cell based high-throughput screening.
Identification of novel inhibitors of M. tuberculosis growth using whole cell based high-throughput screening.
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DOI：
10.1021/cb300151m
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发表时间：
2012-08-17
影响因子：
4
通讯作者：
Hung, Deborah T.
中科院分区：
文献类型：
--
作者：
Stanley, Sarah A.;Grant, Sarah Schmidt;Kawate, Tomohiko;Iwase, Noriaki;Shimizu, Motohisa;Wivagg, Carl;Silvis, Melanie;Kazyanskaya, Edward;Aquadro, John;Golas, Aaron;Fitzgerald, Michael;Dai, Huanqin;Zhang, Lixin;Hung, Deborah T.
尽管迫切需要新的抗结核药物，但几乎没有。为了应对新出现的耐药性问题，将需要抑制新靶点的结构独特的化学实体。在这里，我们描述了我们的调查使用全细胞筛选的小分子作为一种方法，用于确定新的抑制剂，靶向结核分枝杆菌药物发现的新途径。我们发现，使用模型分枝杆菌物种进行初步筛选可能会限制识别对M有效的新抑制剂的潜力。结核此外，我们确认了开发反映体内生物学的体外测定条件的重要性，以最大化来自可能具有体内活性的全细胞筛选的命中比例。最后，我们描述了两个新的抑制剂的目标步骤，在M。结核菌细胞壁生物合成。第一个是一种新的苯并咪唑，其靶向分枝杆菌膜蛋白大3（MmpL 3），一种拟议的细胞壁分枝杆菌酸转运蛋白。第二种是硝基三唑，可抑制十异戊二烯磷酰基-β-d-核糖2′-差向异构酶（DprE 1），这是细胞壁生物合成所需的差向异构酶。这些蛋白质都是通过正向化学遗传学使用抗性产生结合基因组测序鉴定的少数新靶标。这表明，目前用于筛选和目标识别的方法可能会导致目标发现的偏差，应探索替代方法。
Despite the urgent need for new antitubercular drugs, few are on the horizon. To combat the problem of emerging drug resistance, structurally unique chemical entities that inhibit new targets will be required. Here we describe our investigations using whole cell screening of a diverse collection of small molecules as a methodology for identifying novel inhibitors that target new pathways for Mycobacterium tuberculosis drug discovery. We find that conducting primary screens using model mycobacterial species may limit the potential for identifying new inhibitors with efficacy against M. tuberculosis. In addition, we confirm the importance of developing in vitro assay conditions that are reflective of in vivo biology for maximizing the proportion of hits from whole cell screening that are likely to have activity in vivo. Finally, we describe the identification and characterization of two novel inhibitors that target steps in M. tuberculosis cell wall biosynthesis. The first is a novel benzimidazole that targets mycobacterial membrane protein large 3 (MmpL3), a proposed transporter for cell wall mycolic acids. The second is a nitro-triazole that inhibits decaprenylphosphoryl-β-d-ribose 2′-epimerase (DprE1), an epimerase required for cell wall biosynthesis. These proteins are both among the small number of new targets that have been identified by forward chemical genetics using resistance generation coupled with genome sequencing. This suggests that methodologies currently employed for screening and target identification may lead to a bias in target discovery, and that alternative methods should be explored.
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影响因子：
168.9
作者：
Ma, Zhenkun;Lienhardt, Christian;Wang, Xiexiu
通讯作者：
Wang, Xiexiu
影响因子：
4.9
作者：
Brooks, JV;Furney, SK;Orme, IM
通讯作者：
Orme, IM
影响因子：
2.9
作者：
Kwon-Chung, Kyung J.;Sugui, Janyce A.
通讯作者：
Sugui, Janyce A.
影响因子：
3.6
作者：
MULLBACHER, A;WARING, P;EICHNER, RD
通讯作者：
EICHNER, RD
DOI：
10.1016/j.tube.2009.07.006
发表时间：
2009-09
期刊：
Tuberculosis (Edinburgh, Scotland)
影响因子：
--
作者：
Maddry JA;Ananthan S;Goldman RC;Hobrath JV;Kwong CD;Maddox C;Rasmussen L;Reynolds RC;Secrist JA 3rd;Sosa MI;White EL;Zhang W
通讯作者：
Zhang W