量子点介孔复合纳米荧光探针靶向筛选中草药中的抗肿瘤Hsp90抑制剂

Inhibition of heat shock protein (Hsp90) can regulate a variety of tumor signal channel to inhibit proliferation and metastases, thereby, has become the focus of anti-tumor research, and screening Hsp90 inhibitors from herbs contains great value. However, the composition of Chinese herbal medicine is very complex, and which would seriously interfere screening for active molecular. So, presently screening technology is very difficult to play the role of. How to realize accurately, efficient, rapid screening of the active substance from the Chinese herbal medicine highly complex compositions has become an urgent problem to solve. The project around this key problem would design and preparation of high specific surface area mesoporous nano-materials to fix target protein Hsp90 by the surface modification. The quantum dots as the tracer were bond onto the surface of mesoporous nano-materials to composite mesoporous nanomaterial fluorescent probe based on quantum dots, targeting screening Hsp90 inhibitor from the Chinese herbal medicines. Through the cell imaging of the probe-active molecules complexes, the effect of active molecules to cells could be studied, and inhibition of Hsp90 activity and pharmacological experiments would validate its activity. Our project not only can greatly improve the accumulation ability of active molecule, but also has the advantages of real-time monitoring. More importantly, it could exclude the interference of the non active molecules in Chinese herbal medicine, which can become simple and reliable platform for high-throughput screening.

抑制热休克蛋白(Hsp90)可调控多种肿瘤信号通路，从而抑制肿瘤的增殖、转移，已成为抗肿瘤研究的热点，从中草药中筛选Hsp90抑制剂蕴藏着巨大的价值。然而，中草药成分异常复杂，严重干扰活性分子筛选，目前的筛选技术很难发挥作用。如何实现在成分高度复杂的中草药中准确、高效、快速筛选、富集活性分子已成为急需解决的问题。本项目围绕这一关键问题，拟设计和制备巨大比表面的介孔纳米材料，表面进行修饰固定靶蛋白Hsp90；制备优异荧光性能的量子点作为示踪剂复合到介孔纳米材料表面；组成量子点介孔复合纳米荧光探针，靶向筛选中草药中的Hsp90抑制剂；通过对筛选后的探针－活性分子复合物进行细胞成像，研究活性分子对细胞的作用，以及进行Hsp90抑制率测定、药理实验等验证活性。该技术不仅大大提高活性分子富集能力，且具有荧光实时监测的优势，更重要的是能够排除中草药中非活性分子的干扰，从而可以成为可靠的高通量筛选平台。

我国传统中医药具有悠久的历史, 现有证据表明中药的多种成分可作用于多个疾病靶点，在中草药中找寻天然的活性成分（群）越来越成为近年来医药科学研究的一个热点。然而，植物粗提物或者中药水煎物中成分十分复杂，严重干扰活性分子群的筛选，并且还存在着不能即时反应筛选结果、耗时长、需要进行繁琐的样品预处理以及不能有效富集活性分子的问题。另一方面，生物体液中基质复杂，具有很强的背景信号，与目标物信号相互干扰，因此造成中药功效物质活性导向的靶标蛋白识别变得异常困难。本项目围绕这些关键问题，发展了“荧光成像-配体垂钓-Imaging Screening and Fluorescent ligand fishing”技术，结合荧光成像和配体垂钓固相萃取，实现了在活性分子筛选过程中原位成像分析，应用于雷公藤、姜黄、夏枯草等中药复杂体系中识别、富集、靶向筛选活性分子，发现、筛选得到抗肿瘤HSP90活性分子20余种；系统地建立了中药复杂体系中有关活性分子识别、富集、分离、追踪检测新技术及新方法；系统地发展了一系列灵敏度高、特异性强的生物传感器，解决了生物体液中基质复杂，具有复杂的背景信号，与目标物信号相互干扰的研究难点，应用于肿瘤相关标志物血小板衍生长因子PDGF-BB、miRNA、端粒酶活性的传感研究。这些发现不仅为相关中药的化学物质基础及作用机理提供了重要起始分子，而且为复杂生物基质活性分子及靶标蛋白的发掘研究提供了有效策略与方法。