Gametocytocidal screen identifies novel chemical classes with Plasmodium falciparum transmission blocking activity.
Gametocytocidal screen identifies novel chemical classes with Plasmodium falciparum transmission blocking activity.
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DOI：
10.1371/journal.pone.0105817
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Tripathi AK
中科院分区：
文献类型：
--
作者：
Sanders NG;Sullivan DJ;Mlambo G;Dimopoulos G;Tripathi AK
发现传播阻断化合物是消除和根除疟疾所必需的重要干预策略。迄今为止，只有少数药物可以抑制配子细胞的发育，从而抑制从蚊子到人类宿主的传播。这种限制很大程度上是由于缺乏易于适应高通量的筛选分析，因为有多个孵育步骤或对高配子体细胞的要求。在这里，我们报告了使用简单而强大的SYBR Green I为基础的DNA分析发现的具有杀配子体活性的新化合物。我们的实验利用雄性配子体的鞭毛步骤和一个背景抑制因子，通过增加活寄生虫的信号和减少死寄生虫的信号来掩盖死细胞的染色，从而达到健康的信噪比。通过确定鞭毛对荧光信号的贡献并使用适当的截止值，我们能够筛选杀配子体化合物。经过实验验证和优化，我们筛选了FDA批准的约1500种化合物的药物库，以及400种化合物MMV疟疾盒，并鉴定了44种具有亚至低微摩尔ic50的配子体杀伤化合物。具有杀配子体活性的化合物的主要类别包括与胆碱结构相似的季铵盐化合物，与喹啉和吡啶相似的吖啶类化合物，以及抗抑郁、抗肿瘤和驱虫化合物。在膜喂养试验中，顶级候选药物显示出接近完全的传输阻断。该试验简单、可重复性好，且在低配子细胞水平下显示出稳健的z因子值，使其适用于HTS用于鉴定新型和有效的配子细胞杀伤化合物。
Discovery of transmission blocking compounds is an important intervention strategy necessary to eliminate and eradicate malaria. To date only a small number of drugs that inhibit gametocyte development and thereby transmission from the mosquito to the human host exist. This limitation is largely due to a lack of screening assays easily adaptable to high throughput because of multiple incubation steps or the requirement for high gametocytemia. Here we report the discovery of new compounds with gametocytocidal activity using a simple and robust SYBR Green I- based DNA assay. Our assay utilizes the exflagellation step in male gametocytes and a background suppressor, which masks the staining of dead cells to achieve healthy signal to noise ratio by increasing signal of viable parasites and subtracting signal from dead parasites. By determining the contribution of exflagellation to fluorescent signal and using appropriate cutoff values, we were able to screen for gametocytocidal compounds. After assay validation and optimization, we screened an FDA approved drug library of approximately 1500 compounds, as well as the 400 compound MMV malaria box and identified 44 gametocytocidal compounds with sub to low micromolar IC50s. Major classes of compounds with gametocytocidal activity included quaternary ammonium compounds with structural similarity to choline, acridine-like compounds similar to quinacrine and pyronaridine, as well as antidepressant, antineoplastic, and anthelminthic compounds. Top drug candidates showed near complete transmission blocking in membrane feeding assays. This assay is simple, reproducible and demonstrated robust Z-factor values at low gametocytemia levels, making it amenable to HTS for identification of novel and potent gametocytocidal compounds.
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影响因子：
3.7
作者：
Coulibaly B;Zoungrana A;Mockenhaupt FP;Schirmer RH;Klose C;Mansmann U;Meissner PE;Müller O
通讯作者：
Müller O
影响因子：
3.7
作者：
Lelièvre J;Almela MJ;Lozano S;Miguel C;Franco V;Leroy D;Herreros E
通讯作者：
Herreros E
影响因子：
7.5
作者：
Fernandez-Calienes, Ayme;Pellon, Rolando;Rojas, Lazara
通讯作者：
Rojas, Lazara
影响因子：
5.2
作者：
D'Alessandro, Sarah;Silvestrini, Francesco;Taramelli, Donatella
通讯作者：
Taramelli, Donatella
影响因子：
4.9
作者：
Downey, Autumn S.;Chong, Curtis R.;Sullivan, David J.
通讯作者：
Sullivan, David J.