Division of Preclinical Innovation, National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, 20850, USA.
Department of Microbiology and Immunology, Columbia University Irving Medical Center, New York, NY, 10032, USA.
Sci Rep. 2021 Jan 22;11(1):2121. doi: 10.1038/s41598-021-81486-z.
The spread of Plasmodium falciparum parasites resistant to most first-line antimalarials creates an imperative to enrich the drug discovery pipeline, preferably with curative compounds that can also act prophylactically. We report a phenotypic quantitative high-throughput screen (qHTS), based on concentration-response curves, which was designed to identify compounds active against Plasmodium liver and asexual blood stage parasites. Our qHTS screened over 450,000 compounds, tested across a range of 5 to 11 concentrations, for activity against Plasmodium falciparum asexual blood stages. Active compounds were then filtered for unique structures and drug-like properties and subsequently screened in a P. berghei liver stage assay to identify novel dual-active antiplasmodial chemotypes. Hits from thiadiazine and pyrimidine azepine chemotypes were subsequently prioritized for resistance selection studies, yielding distinct mutations in P. falciparum cytochrome b, a validated antimalarial drug target. The thiadiazine chemotype was subjected to an initial medicinal chemistry campaign, yielding a metabolically stable analog with sub-micromolar potency. Our qHTS methodology and resulting dataset provides a large-scale resource to investigate Plasmodium liver and asexual blood stage parasite biology and inform further research to develop novel chemotypes as causal prophylactic antimalarials.
疟原虫对大多数一线抗疟药物产生抗药性，这就迫切需要丰富药物研发管道，最好是使用既能治疗又能预防的治愈化合物。我们报告了一种基于浓度反应曲线的表型定量高通量筛选（qHTS），旨在鉴定对疟原虫肝脏和无性血阶段寄生虫有活性的化合物。我们的 qHTS 筛选了超过 450,000 种化合物，在 5 到 11 个浓度范围内进行了测试，以评估它们对恶性疟原虫无性血阶段的活性。然后，对活性化合物进行独特结构和类药性的过滤，随后在伯氏疟原虫肝脏阶段测定中进行筛选，以鉴定新的双活性抗疟化学型。噻二嗪和嘧啶氮杂环庚烷化学型的化合物随后被优先用于耐药性选择研究，导致恶性疟原虫细胞色素 b 中的明确突变，这是一个已验证的抗疟药物靶点。噻二嗪化学型进行了初步的药物化学研究，得到了一种具有亚毫摩尔效力的代谢稳定类似物。我们的 qHTS 方法和由此产生的数据集为研究疟原虫肝脏和无性血阶段寄生虫生物学提供了一个大规模资源，并为开发新的化学型作为因果预防性抗疟药物提供了进一步的研究信息。
