双靶向AMPK和ZIPK的新型抗宫颈癌活性化合物的优化设计和凋亡机制

Carcinogenesis is a complex process involved in multiple factors, genes and steps; therefore, using the systems biology network to seek potential drug targets and design novel anti-tumor compounds based upon these targets might reveal new direction of small molecule compounds in cancer. According to the core apoptotic signaling pathways from the Naïve Bayesian model network, we found that AMPK was in the central zone of the predicted apoptotic kinase network, working jointly with ZIPK to regulate most of core apoptotic kinase-related signaling pathways. Thus, based on the structures of AMPK and ZIPK, we used a series of methods such as molecular docking, dynamics stimulation and chemical synthesis to get an indole ketone-like compound AD008, which could bind AMPK and ZIPK very well. In addition, we carried out MTT, microscopy observation, flow cytometry, PI, western blot and siRNA to show that AD008 could induce HeLa cell apoptosis. Thus, in this study, we will undertake in-depth optimization and design of the already acquired structure of AD008, expecting that the optimized compound may improve anti-proliferative activity and apoptosis-inducing effect in cervical cancer. Moreover, we will explore more new signaling pathways of the optimized compound-induced apoptosis in vitro and in vivo, especially highlighting education of new apoptotic mechanisms of the optimized compound targeting both AMPK and ZIPK. Together, these findings will provide a new clue for further exploiting more anti-tumor dual-target candidate compounds for future cancer therapy.

肿瘤发生是一个多因素参与、多基因改变、多步骤的复杂过程。利用系统生物学网络寻找潜在靶点，设计新型靶向抗肿瘤活性化合物，是肿瘤药物的重要方向。在前期朴素贝叶斯模型构建网络研究凋亡的工作中，我们发现AMPK处于预测的凋亡激酶的网络核心，并联同ZIPK共同调控凋亡激酶核心通路。基于AMPK和ZIPK的结构通过分子对接、动力学模拟和药物合成等手段得到了一个吲哚酮类化合物AD008。用MTT法、形态学观察、流式细胞术、Western blot 和siRNA等手段发现AD008对AMPK与ZIPK均具有靶向性结合并通过激活AMPK和ZIPK来诱导HeLa细胞凋亡。因此，我们在本项目中拟将已获得AD008的结构再优化设计，以期优化后化合物能提高对宫颈癌的抗增殖活性和诱导凋亡的效果。我们还研究它在体内外诱导凋亡的新通路，并重点阐述它双靶向AMPK和ZIPK的凋亡新机制，为新型抗宫颈癌候选药物研究提供基础。

肿瘤发生是一个多因素参与、多基因改变、多步骤的复杂过程。利用系统生物学网络寻找潜在靶点，设计新型靶向抗肿瘤活性化合物，是肿瘤药物的重要方向。我们在前期朴素贝叶斯模型构建网络研究凋亡的工作中，发现AMPK处于预测的凋亡激酶的网络核心，并联同ZIPK共同调控凋亡激酶核心通路，因此推测其可作为两个潜在的凋亡靶点被运用于肿瘤药物开发。在本项目的支持下，我们进一步开展了对先导化合物的结构优化和活性筛选，并通过宫劲癌细胞系和异种移植瘤小鼠模型，从细胞、分子和整体动物水平多方面的明确了新合成的AMPK/ZIPK双靶向小分子激动剂在体外及体内的抗肿瘤功效及其诱导肿瘤细胞凋亡的机制。.结果表明， BL-AD008具有较好的抗宫颈癌细胞增殖活性，并可以通过死亡受体通路和线粒体通路促进细胞凋亡。且由BL-AD008促进的凋亡可能是通过对AMPK和ZIPK的同时影响造成的，体内实验同样证明了BL-AD008拥有着极佳的抗肿瘤活性与通过靶向AMPK和ZIPK来促进凋亡的能力。这些研究结果表明了在系统生物学网络中确认自噬关键激酶靶点AMPK和ZIPK的能力，并提供了一个双靶向小分子化合物（BL-AD008）作为在宫颈癌治疗中调控细胞凋亡的潜在药物。.在本项目研究经费的资助下，项目负责人以第一作者或通讯作者在本领域权威期刊上共发表了SCI论文16篇，并获得省部级二等奖2项，做特邀报告一次，参加其他学术会议3次。