Cheng Ken Chih-Chien, Cao Shugeng, Raveh Avi, MacArthur Ryan, Dranchak Patricia, Chlipala George, Okoneski Matthew T, Guha Rajarshi, Eastman Richard T, Yuan Jing, Schultz Pamela J, Su Xin-Zhuan, Tamayo-Castillo Giselle, Matainaho Teatulohi, Clardy Jon, Sherman David H, Inglese James
National Center of Advancing Translational Sciences, National Institutes of Health , Rockville, Maryland 20850, United States.
Department of Pharmaceutical Sciences, Daniel K. Inouye College of Pharmacy, University of Hawai'i at Hilo , Hilo, Hawaii 96720, United States.
J Nat Prod. 2015 Oct 23;78(10):2411-22. doi: 10.1021/acs.jnatprod.5b00489. Epub 2015 Oct 14.
Methods to identify the bioactive diversity within natural product extracts (NPEs) continue to evolve. NPEs constitute complex mixtures of chemical substances varying in structure, composition, and abundance. NPEs can therefore be challenging to evaluate efficiently with high-throughput screening approaches designed to test pure substances. Here we facilitate the rapid identification and prioritization of antimalarial NPEs using a pharmacologically driven, quantitative high-throughput-screening (qHTS) paradigm. In qHTS each NPE is tested across a concentration range from which sigmoidal response, efficacy, and apparent EC50s can be used to rank order NPEs for subsequent organism reculture, extraction, and fractionation. Using an NPE library derived from diverse marine microorganisms we observed potent antimalarial activity from two Streptomyces sp. extracts identified from thousands tested using qHTS. Seven compounds were isolated from two phylogenetically related Streptomyces species: Streptomyces ballenaensis collected from Costa Rica and Streptomyces bangulaensis collected from Papua New Guinea. Among them we identified actinoramides A and B, belonging to the unusually elaborated nonproteinogenic amino-acid-containing tetrapeptide series of natural products. In addition, we characterized a series of new compounds, including an artifact, 25-epi-actinoramide A, and actinoramides D, E, and F, which are closely related biosynthetic congeners of the previously reported metabolites.
鉴定天然产物提取物（NPEs）中生物活性多样性的方法不断发展。NPEs是结构、组成和丰度各异的化学物质的复杂混合物。因此，使用旨在测试纯物质的高通量筛选方法对NPEs进行有效评估可能具有挑战性。在此，我们使用药理学驱动的定量高通量筛选（qHTS）范式，促进抗疟NPEs的快速鉴定和优先级排序。在qHTS中，每个NPE在一个浓度范围内进行测试，从该浓度范围可以使用S形响应、效力和表观EC50对NPEs进行排序，以便随后进行生物体再培养、提取和分级分离。使用来自不同海洋微生物的NPE文库，我们从数千个使用qHTS测试的样品中观察到两种链霉菌属提取物具有强大的抗疟活性。从两个系统发育相关的链霉菌物种中分离出七种化合物：从哥斯达黎加采集的巴氏链霉菌和从巴布亚新几内亚采集的邦古拉链霉菌。其中我们鉴定出放线酰胺A和B，它们属于天然产物中异常复杂的含非蛋白质ogenic氨基酸的四肽系列。此外，我们还表征了一系列新化合物，包括一种假象物25-表-放线酰胺A以及放线酰胺D、E和F，它们是先前报道的代谢物的密切相关生物合成同源物。
