Lucantoni Leonardo, Fidock David A, Avery Vicky M
Discovery Biology, Eskitis Institute for Drug Discovery, Griffith University, Nathan, Queensland, Australia.
Department of Microbiology and Immunology, Columbia University College of Physicians and Surgeons, New York, New York, USA Division of Infectious Diseases, Department of Medicine, Columbia University College of Physicians and Surgeons, New York, New York, USA.
Antimicrob Agents Chemother. 2016 Mar 25;60(4):2097-107. doi: 10.1128/AAC.01949-15. Print 2016 Apr.
The discovery of new antimalarial drugs able to target both the asexual and gametocyte stages ofPlasmodium falciparumis critical to the success of the malaria eradication campaign. We have developed and validated a robust, rapid, and cost-effective high-throughput reporter gene assay to identify compounds active against late-stage (stage IV and V) gametocytes. The assay, which is suitable for testing compound activity at incubation times up to 72 h, demonstrates excellent quality and reproducibility, with averageZ' values of 0.85 ± 0.01. We used the assay to screen more than 10,000 compounds from three chemically diverse libraries. The screening outcomes highlighted the opportunity to use collections of compounds with known activity against the asexual stages of the parasites as a starting point for gametocytocidal activity detection in order to maximize the chances of identifying gametocytocidal compounds. This assay extends the capabilities of our previously reported luciferase assay, which tested compounds against early-stage gametocytes, and opens possibilities to profile the activities of gametocytocidal compounds over the entire course of gametocytogenesis.
发现能够靶向恶性疟原虫无性繁殖阶段和配子体阶段的新型抗疟药物对于疟疾根除运动的成功至关重要。我们开发并验证了一种强大、快速且经济高效的高通量报告基因检测方法，以鉴定对晚期（IV期和V期）配子体有活性的化合物。该检测方法适用于在长达72小时的孵育时间内测试化合物活性，显示出优异的质量和可重复性，平均Z'值为0.85±0.01。我们使用该检测方法对来自三个化学性质不同的文库的一万多种化合物进行了筛选。筛选结果突出了利用对寄生虫无性繁殖阶段具有已知活性的化合物库作为检测杀配子体活性的起点的机会，以便最大限度地提高鉴定杀配子体化合物的可能性。该检测方法扩展了我们之前报道的针对早期配子体测试化合物的荧光素酶检测方法的能力，并为在配子体发生的整个过程中分析杀配子体化合物的活性开辟了可能性。
