Identification of Small Molecule Inhibitors of Clostridium perfringens ε-Toxin Cytotoxicity Using a Cell-Based High-Throughput Screen.
Identification of Small Molecule Inhibitors of Clostridium perfringens ε-Toxin Cytotoxicity Using a Cell-Based High-Throughput Screen.
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DOI：
10.3390/toxins2071825
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发表时间：
2010-07-01
期刊：
影响因子：
4.2
通讯作者：
McClain MS
中科院分区：
文献类型：
--
作者：
Lewis M;Weaver CD;McClain MS
产气荚膜梭菌肠毒素是一种选择性因子，可导致严重的、通常致命的肠毒血症，其特征在于心脏、肺、肾和脑水肿。该毒素被认为是寡聚成孔毒素。目前，没有有效的治疗方法来对抗暴露个体中毒素的细胞毒性活性。使用稳健的基于细胞的高通量筛选（HTS）测定，我们筛选了151，616-化合物文库抑制ε-毒素诱导的细胞毒性的能力。通过添加刃天青以检测存活细胞中的代谢活性来评估暴露于毒素的MDCK细胞的存活。这个画面的命中率是0.6%。在一式三份地对每个命中物进行二次筛选并进行测定以消除假阳性之后，我们集中于三种结构不同的化合物：N-环烷基苯甲酰胺、呋喃并[2，3-B]喹啉和6 H-蒽并[1，9-cd]异恶唑。这三种化合物似乎都不能抑制毒素与细胞的结合或毒素形成寡聚复合物的能力。另外的测定表明，两种抑制性化合物抑制ε-毒素诱导的MDCK细胞对碘化丙啶的透化。此外，这两种化合物对毒素预处理的细胞表现出抑制作用。通过高通量筛选鉴定的抑制剂之一的结构类似物进行了分析，并提供了初始的结构-活性数据。这些化合物应该作为进一步结构-活性改进的基础，这可能导致开发有效的抗ε-毒素治疗剂。
The Clostridium perfringens epsilon toxin, a select agent, is responsible for a severe, often fatal enterotoxemia characterized by edema in the heart, lungs, kidney, and brain. The toxin is believed to be an oligomeric pore-forming toxin. Currently, there is no effective therapy for countering the cytotoxic activity of the toxin in exposed individuals. Using a robust cell-based high-throughput screening (HTS) assay, we screened a 151,616-compound library for the ability to inhibit ε-toxin-induced cytotoxicity. Survival of MDCK cells exposed to the toxin was assessed by addition of resazurin to detect metabolic activity in surviving cells. The hit rate for this screen was 0.6%. Following a secondary screen of each hit in triplicate and assays to eliminate false positives, we focused on three structurally-distinct compounds: an N-cycloalkylbenzamide, a furo[2,3-b]quinoline, and a 6H-anthra[1,9-cd]isoxazol. None of the three compounds appeared to inhibit toxin binding to cells or the ability of the toxin to form oligomeric complexes. Additional assays demonstrated that two of the inhibitory compounds inhibited ε-toxin-induced permeabilization of MDCK cells to propidium iodide. Furthermore, the two compounds exhibited inhibitory effects on cells pre-treated with toxin. Structural analogs of one of the inhibitors identified through the high-throughput screen were analyzed and provided initial structure-activity data. These compounds should serve as the basis for further structure-activity refinement that may lead to the development of effective anti-ε-toxin therapeutics.
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影响因子：
2.6
作者：
Minami, J;Katayama, S;Okabe, A
通讯作者：
Okabe, A
DOI：
10.1016/j.bbrc.2003.08.112
发表时间：
2003-10-10
影响因子：
3.1
作者：
Burnett, JC;Schmidt, JJ;Bavari, S
通讯作者：
Bavari, S
影响因子：
3.1
作者：
McClain, Mark S.;Cover, Timothy L.
通讯作者：
Cover, Timothy L.
影响因子：
5.8
作者：
Makarenkov, Vladimir;Kevorkov, Dmytro;Nadon, Robert
通讯作者：
Nadon, Robert
影响因子：
--
作者：
Brideau, C;Gunter, B;Liaw, A
通讯作者：
Liaw, A