基于苦参碱结构的新骨架衍生物的合成、抗肿瘤构效关系和作用机理研究

Cancer has become a global issue with its high mortality and great remedial difficulty, and been threatening human health and life. Thus, it is imperative to seek and develop novel anticancer drugs. Based on the previous research of matrin in our group, we will design and synthesize a series of matrine derivatives with novel skeleton structures in the project. Necessarily, we will also study the synthesis methods and optimize relative parameters, and use spectroscopy analysis to characterize the structures of target compounds. In vivo antitumor activities of target compounds will be evaluated by pharmacological experimental models such as Ehrlich's ascites carcinoma and so on. The structure-activity relationships will be analyzed by computer aided methods. Inhibition effects of target compounds on proliferation, adhesion, infiltration and metastasis of tumor cells and induced apoptosis of tumor sells will be researched in cellular and molecular levels. The regulation mechanism of tumor-relative genes for matrine derivatives will be further studied. Finally, the effect mechanism of the compounds will be clarified. From the combination pattern of the targeted protein and the compounds in molecular docking, we will get the interaction relationships between anti-tumor compounds and the drug target. The research results will establish a foundation for designing more potent anticancer candidates and developing new drugs.

癌症已成为全球性的问题，死亡率高，治疗难度大，严重地威胁着人类的生命健康。因此，寻找和开发新型抗癌药物势在必行。本项目在已有对苦参碱研究工作的基础上，设计合成一系列新骨架结构的衍生物。探索合成方法，优化合成参数，并采用波谱分析手段表征目标化合物的结构。采用艾氏腹水癌等药理实验模型进行体内抗肿瘤药理活性测定，结合计算机辅助手段，研究目标化合物的构效关系。在细胞和分子水平上，研究目标化合物对肿瘤细胞的增值抑制、粘附与浸润转移抑制、诱导凋亡等作用，并进一步研究苦参碱衍生物对肿瘤相关基因的调控机制，明确目标化合物的作用机理。对目标化合物进行分子对接研究，从与靶标蛋白的结合方式上，明确化合物活性与靶标相互作用的关系。研究结果将为进一步设计有效地抗肿瘤药物候选物，开发新药建立基础。

设计并完成合成161个三个系列苯并-γ-吡喃酮苦参碱新骨架衍生物、14位取代的苦参碱衍生物和开环的苦参碱衍生物，并用波谱分析手段表征了目标化合物。对合成的目标化合物进行体外抗肿瘤药理活性测定研究，选择H460、MCF-7、SGC-7901、Bel-7402等不同的肿瘤细胞系测定了抑制活性，计算它们的IC50值。对部分化合物进行了体内抗肿瘤活性研究，包括H22荷瘤小鼠肿瘤的抑制作用和A549小鼠肺癌的抑制作用。获得了4个高活性的化合物，ZS-06、ZS-07、ZS-10和ZS-12，肿瘤细胞实验结果表明它们的IC50小于10μM。抗肿瘤作用机理研究表明，化合物5i在肺癌细胞中能通过上调P27、下调CDK4和细胞周期蛋白D1以及抑制PI3K/Akt/mTOR通路诱导G1细胞周期的阻滞和自我吞噬；化合物CH6在肝癌细胞Bel-7402 和 HepG2中通过上调P21、P27和E-钙粘素蛋白以及下调N-钙粘素蛋白诱导G1细胞周期的阻滞和抑制细胞迁移；YF-18 通过下调Skp2和上调它的底物、P27和E-钙粘素蛋白剂量依赖性的诱导肺癌细胞G2/M细胞周期的阻滞和抑制迁移。目标化合物YF-8与细胞周期调节靶标和细胞凋亡靶标对接模拟，对接结果显示，YF-8与CDK2之间的形成范德华力、静电作用、疏水作用以及氢键作用，才使得化合物可以与靶标紧密的结合，从而抑制其活性。