基于cmER靶点的新疆阿魏树脂中倍半萜香豆素类抗宫颈癌活性成分及构效关系研究

Cervical cancer is one of the most common malignant tumors in the female reproductive system, and the drug resistance of cervical cancer cells is the main problem of its clinical treatment. Therefore, it’s urgent to discover new structurally specific anti-cervical cancer drugs that act on new targets. Ferula sinkiangensis is a traditional medicine of Uyghur. Our team has isolated a series of sesquiterpene coumarins (SCs) with novel structures from its resin, and their anti-cervical cancer activities are even better than the clinical drug 5-fluorouracil (5-FU). Further studies have revealed that SCs may play an anticancer role by inhibiting the cell membrane-bound estrogen receptor (cmER). This project is intended to apply HPLC, LC-LTQ-Orbitrap and UPC2, combined with screening of anticancer activity in vitro and nude mouse xenograft model in vivo, to quickly identify and obtain SCs with novel structures and satisfactory activity. At the same time, indirect fluorescence staining, flow cytometry and Western blot are applied to verify the effects of active ingredients on cmER target, and the anticancer structure-activity relationship of SCs was further analyzed by the combination of virtual screening technology and screening of anticancer activity in vitro. The aim of this study is to discover anti-cervical cancer lead compounds that act on new target, and to provide a new strategy to rationally develop and utilize traditional ethnic medicine.

宫颈癌是女性生殖系统最常见的恶性肿瘤之一，其耐药性是一直临床治疗的主要难题。因此，发现结构特异的作用于新靶点的抗宫颈癌药物迫在眉睫。新疆阿魏为维族传统用药，课题组前期从新疆阿魏树脂中分离得到一系列结构新颖的倍半萜香豆素类化合物（SCs），通过药理实验发现其抗宫颈癌活性甚至优于临床用药5-氟尿嘧啶（5-FU），进一步研究显示SCs可能通过抑制细胞膜结合的雌激素受体（cmER）发挥抗癌作用。本项目拟应用HPLC-PDA、LC-LTQ-Orbitrap及UPC2，结合体外抗癌活性筛选和体内裸鼠移植瘤模型，快速识别和获得结构新颖和活性良好的SCs。同时应用间接免疫荧光染色、流式细胞术及Western blot技术验证活性成分对cmER靶标的作用，并结合虚拟筛选技术和体外抗癌活性筛选结果进而深入剖析SCs的抗癌构效关系，发现具有新颖作用靶点的抗宫颈癌先导化合物，并为合理开发利用传统民族药提供新思路。

宫颈癌是女性生殖系统最常见的恶性肿瘤之一，其耐药性是一直临床治疗的主要难题。因此，发现结构特异的作用于新靶点的抗宫颈癌药物迫在眉睫。新疆阿魏为维族传统用药，课题组前期研究发现新疆阿魏中的倍半萜香豆素类化合物（SCs）体外抗宫颈癌细胞HeLa 增殖的效果优于临床用药5-氟尿嘧啶（5-FU），且可能作用于不同的抗癌靶点。基于此，本项目应用HPLC-PDA、LC-LTQ-Orbitrap从目标部位中快速识别了84个SCs，同时应用各种色谱光谱学手段从中分离鉴定了107个化合物（新化合物27个），包括59个SCs（新化合物15个，2个为罕见的倍半萜香豆素过氧化物）。体外活性筛选结果表明SCs为其抗肿瘤活性成分，59个SCs对HeLa细胞均展现出了不同程度的增殖抑制作用，基于药效团模型搜索显示cmER为其作用靶点，结合体外活性筛选结果及分子对接结果，分析了SCs的体外抗肿瘤活性构效关系。最后使用流式细胞术、Western blot等技术证明，SCs通过抑制cmER靶点，激活下游MAPK通路，诱导HeLa细胞凋亡及G0/G1期阻滞，从而发挥其抗肿瘤作用。本项目最终发现了一个具有新颖作用靶点的抗宫颈癌先导化合物Sinkiangenol E，并为合理开发利用传统民族药提供了新思路。