基于c-Met靶点的三宝木属壮药中酪氨酸激酶抑制剂的发现及机制研究

Tyrosine kinases play a very important role in the process of tumor occurrence and development. Research and development of antitumor drugs has become the hot topics based on Tyrosine kinase targets. Natural products with novel structures, such as diterpenoids and alkaloids, had been isolated from plants of the genus Trigonostemon, but their antitumor mechanism had not been further researched. Further research found alcohol extracts and some indole alkaloids having inhibitory activity for c-Met tyrosine kinase. We will herein take two or three species of the genus Trigonostemon in Guangxi province as research objects and do a detailed study, including rapid detection and target separation of the target compounds with tyrosine kinase inhibitory activity based on target enzyme−ultraviolet spectrum−HPLC-MS technology. Meantimes modern pharmacological models and methods are used to investigate antitumor mechanism and structure-activity relationship at cells, molecules and animal levels respectively. This study will not only provides new idea to discover antitumor leading compounds but also have significant sense and practical value for resource development of traditional minority medicine and research and development of tyrosine kinase inhibitors with independent intellectual property rights in our country.

酪氨酸激酶在肿瘤的发生、发展过程中起着非常重要的作用，以酪氨酸激酶为靶点进行药物研发已成为国际上抗肿瘤药物研究的热点。前期研究发现三宝木属植物含有二萜和生物碱等成分，结构新颖且多具体外抗肿瘤活性，其中三宝木中瑞香烷型二萜trigonothyrin C（含9,12,1′′′-环氧环）对c-Met酪氨酸激酶具有较强的抑制活性，预实验发现其醇提物和新分得的吲哚生物碱也有一定活性。因此，本项目拟对研究比较薄弱的两三种广西三宝木属植物进行深入研究，基于c-Met靶酶－紫外光谱－液质联用技术，采用活性追踪与在线检测相结合的方法，快速发现和靶向分离具有酪氨酸激酶抑制活性的化合物，采用现代药理模型与方法，从细胞及分子水平探讨其抗肿瘤作用机制和构效关系。本研究不仅为寻找具有抗肿瘤活性的先导化合物提供思路，而且对我国民族药用资源综合开发利用和自主知识产权研制新型酪氨酸激酶抑制剂具有重要意义。

酪氨酸激酶在肿瘤的发生、发展过程中起着非常重要的作用，从民族药植物资源中寻找发现具有抑制酪氨酸激酶的抗肿瘤活性成分，并探讨其抗肿瘤作用机制。本项目选取壮药植物黄花三宝木，采用活性导向结合紫外光谱－液质联用技术，快速发现与靶向分离目标组分或目标化合物；结果从中分离到33个化合物，包括4个新化合物，其中有10个香豆素类成分。特别是发现的先导化合物为2个新的鞣花单宁，表现具有比较突出的抗肿瘤活性，并发现该类成分可以下调pERK表达，抑制细胞ERK磷酸化，诱导胃癌细胞SGC7901 和结直肠癌细胞HCT116 细胞凋亡；说明其分子机制可能与调节ERK信号通路有关，初步认定HHDP基团为其药效团。同时总结了以紫外光谱－液质联用技术靶向分离香豆素的规律，对10个香豆素初步探讨了其构效关系，发现具共轭双键且含酮羰基的异戊烯基（3-methyl-2-oxobut-3-en-1-yl）为药效团，并发现九里香酮能够有效抑制肝癌HepG2细胞迁移。另外运用正交试验的方法，研究并优化了黄花三宝木部萜的超声辅助提取方法。这些为发现抗肿瘤先导化合物和开发出新的高效抗肿瘤药物提供了一定的理论依据和参考。