High-Throughput Screen in Cryptococcus neoformans Identifies a Novel Molecular Scaffold That Inhibits Cell Wall Integrity Pathway Signaling
High-Throughput Screen in Cryptococcus neoformans Identifies a Novel Molecular Scaffold That Inhibits Cell Wall Integrity Pathway Signaling
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DOI：
10.1021/acsinfecdis.5b00111
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发表时间：
2016-01-01
影响因子：
5.3
通讯作者：
Krysan, Damian J.
中科院分区：
文献类型：
--
作者：
Hartland, Kate;Pu, Jun;Krysan, Damian J.
新型隐球菌是最重要的人类真菌病原体之一，然而，50多年来没有开发出新的治疗方法。杀真菌活性对于有效的抗隐球菌剂是至关重要的，因此，我们筛选了361，675个抗隐球菌的分子。使用特异性检测杀真菌活性的腺苷酸激酶释放测定法测定新生菌。一组二次试验缩小了一组命中干扰真菌细胞壁完整性的分子，并确定了三个苯并硫脲类药物具有低的体外哺乳动物毒性和良好的体外抗隐球菌（最低抑制浓度= 4 μ g/mL）。该支架通过细胞壁完整性MAP激酶级联抑制信号传导。结构-活性研究表明，硫代羰基部分是至关重要的活性。遗传和生化数据表明，苯并硫脲类抑制激酶级联上游的信号传导。因此，苯并硫脲似乎是一种有前途的新支架，可用于进一步探索寻找新的抗隐球菌药物。
Cryptococcus neoformans is one of the most important human fungal pathogens; however, no new therapies have been developed in over SO years. Fungicidal activity is crucially important for an effective anticryptococal agent and, therefore, we screened 361,675 molecules against C. neoformans using an adenylate kinase release assay that specifically detects fungicidal activity. A set of secondary assays narrowed the set of hits to molecules that interfere with fungal cell wall integrity and identified three benzothioureas with low in vitro mammalian toxicity and good in vitro anticryptococcal (minimum inhibitory concentration = 4 mu g/mL). This scaffold inhibits signaling through the cell wall integrity MAP kinase cascade. Structure-activity studies indicate that the thiocarbonyl moiety is crucial for activity. Genetic and biochemical data suggest that benzothioureas inhibit signaling upstream of the kinase cascade. Thus, the benzothioureas appear to be a promising new scaffold for further exploration in the search for new anticryptococcal agents.