以“多成分-多靶点”模式构建双向筛选评价体系解析莪术-三棱药对抗肺癌的药效物质

中药药效物质不明是中药现代研究的关键科学问题。目前药效物质筛选多为单向筛选体系（靶点已知、药效物质未知），难以阐释中药多成分-多靶点特性。我们近期研究表明亲和反应靶点稳定技术（DARTS）在对小分子不做改造下可从肿瘤细胞总蛋白中筛选出目标蛋白。本项目拟将DARTS原理与中药药效物质筛选理念相结合，在靶点和药效物质均未知情况下，构建“多成分-多靶点”模式的双向筛选体系研究莪术-三棱药对抗肺癌的药效物质和作用靶点。将药对提取物与肺癌A549细胞总蛋白孵育，用特定蛋白水解酶酶解游离蛋白，有效成分-靶蛋白复合物因对酶稳定被保留，解离超滤获得的复合物，高分辨质谱分别鉴定解离后的有效成分群和靶蛋白群，活性实验验证有效成分群抗肺癌的活性及整合作用，表面等离子体共振及分子对接等技术验证有效成分与作用靶点的对应关系。本项目不仅有助于发现该药对抗肺癌的药效物质，也可弥补中药药效物质目前只能单向筛选的不足。

Discovery of active components was the key scientific problem in the modernization of traditional Chinese medicines. The methods for screening the active components in traditional Chinese medicines were mostly one-way screening system (targets are known, active components are unknown), and it is difficult to explain the characteristic of multiple components-multiple targets of traditional Chinese medicines. The targets were screened from the cancer cell lysates without any modification of small molecules via the affinity response target stabilization technology (DARTS) in our recent study. DARTS was combined with the concept of screening active components in traditional Chinese medicines in this project, a two-way screening system with a “multiple components-multiple targets” model would be established to screen the anti-lung cancer components in Curcumae rhizoma-Sparganii rhizome herbal pair and the targets. The extracts of the herbal pair were incubated with the total protein of A549 cells, pronase was used to digest the free protein, the active component-target complexes were survived due to the better stability. The complexes were dissociated and separated then, the active components and the targets were identified respectively by high resolution mass spectrometry. The anti-lung cancer activities of these components were verified by following pharmacology experiments, the relationship between the active component and the target was verified by surface plasmon resonance and molecular docking techniques. The anti-lung cancer active components and targets could be screened simultaneously in this project, and it also provided the solution for the shortage of the screen method of traditional Chinese medicines.