Department of Nutritional Sciences & Toxicology, University of California, Berkeley, California 94720, USA.
J Nat Prod. 2011 Dec 27;74(12):2545-55. doi: 10.1021/np200673b. Epub 2011 Nov 30.
A high-throughput (HT) paradigm generating LC-MS-UV-ELSD-based natural product libraries to discover compounds with new bioactivities and or molecular structures is presented. To validate this methodology, an extract of the Indo-Pacific marine sponge Cacospongia mycofijiensis was evaluated using assays involving cytoskeletal profiling, tumor cell lines, and parasites. Twelve known compounds were identified including latrunculins (1-4, 10), fijianolides (5, 8, 9), mycothiazole (11), aignopsanes (6, 7), and sacrotride A (13). Compounds 1-5 and 8-11 exhibited bioactivity not previously reported against the parasite T. brucei, while 11 showed selectivity for lymphoma (U937) tumor cell lines. Four new compounds were also discovered including aignopsanoic acid B (13), apo-latrunculin T (14), 20-methoxy-fijianolide A (15), and aignopsane ketal (16). Compounds 13 and 16 represent important derivatives of the aignopsane class, 14 exhibited inhibition of T. brucei without disrupting microfilament assembly, and 15 demonstrated modest microtubule-stabilizing effects. The use of removable well plate libraries to avoid false positives from extracts enriched with only one or two major metabolites is also discussed. Overall, these results highlight the advantages of applying modern methods in natural products-based research to accelerate the HT discovery of therapeutic leads and/or new molecular structures using LC-MS-UV-ELSD-based libraries.
一种高通量（HT）方法，用于生成基于 LC-MS-UV-ELSD 的天然产物文库，以发现具有新生物活性和/或分子结构的化合物，本研究展示了该方法。为了验证该方法，采用涉及细胞骨架谱分析、肿瘤细胞系和寄生虫的测定法，对来自印度-太平洋海绵 Cacospongia mycofijiensis 的提取物进行了评估。鉴定出了 12 种已知化合物，包括拉氏毒素（1-4、10）、菲甲内酯（5、8、9）、mycothiazole（11）、aignopsanes（6、7）和 sacrotride A（13）。化合物 1-5 和 8-11 对寄生虫 T. brucei 表现出以前未报道的活性，而 11 对淋巴瘤（U937）肿瘤细胞系表现出选择性。还发现了 4 种新化合物，包括 aignopsanoic acid B（13）、apo-latrunculin T（14）、20-甲氧基-菲甲内酯 A（15）和 aignopsane 酮（16）。化合物 13 和 16 代表 aignopsane 类的重要衍生物，14 表现出抑制 T. brucei 的作用而不破坏微丝组装，15 表现出适度的微管稳定作用。还讨论了使用可移动微孔板文库来避免仅含有一种或两种主要代谢物的提取物中出现假阳性的问题。总的来说，这些结果强调了在天然产物研究中应用现代方法的优势，可加速基于 LC-MS-UV-ELSD 的文库发现治疗性先导化合物和/或新的分子结构。
