新颖异活性中心芳烃钌(Ⅱ)配合物疏水亲水平衡与靶向作用的调控机制研究

Based on the dual requirements of hydrophilic-hydrophobic balance and molecular targeted therapy, the hydrophobic carbon chain and hydrophilic chelate (ethylenediamine) will be incorporated into the pyrimidine molecule and new classes of the prodrug ligands (FUen) with excellent hydrophobicity and hydrophilicity will be prepared. Then a series of novel arene ruthenium(II) complexes (FUenRu) with different active centers will be synthesized via the reaction of FUen and the dichloro(p-arene)ruthenium dimer ([Ru(p-arene)Cl2]2). Besides, the cytotoxicity of FUen and FUenRu in vitro, their apparent partition coefficients under physiological conditions and molecular recognition interactions with DNA and thymidylate synthase will be investigated by methyl-thiazol-tetrozolium assay, sulforhodamine B assay, spectroscopy technique, electrochemical technique, etc. According to the theoretical evaluation obtained from computational chemistry software packages (Amber, AutoDock and Gaussian), the molecular targeted regulation mechanism of the title arene ruthenium(II) complexes will be analyzed in order to screen out the prodrug ruthenium(II) arene complexes which have synergistic anticancer effects of hydrophilic-hydrophobic balance and molecular targeted regulation, and arene ruthenium(II) complexes will be developed into a kind of novel drug delivery vehicle which possessed excellent anticancer activity and tumor-targeting performance. There is no doubt that above research achievement will provide a novel research approach and valuable regulation mechanism for the joint application of targeted anticancer drug and drug delivery carrier.

基于药物运输的疏水亲水平衡原理以及药物靶向性的双重要求，在嘧啶类抗癌药物分子的N1位引入疏水碳链和亲水螯合基团(乙二胺)，合成一系列兼具水溶性和脂溶性的目标前药配体(FUen)，再与同活性中心的芳烃钌二聚体协配成新颖的异活性中心目标前药配合物(FUenRu)。运用四氮唑盐法、磺酰罗丹明B 蛋白染色法、光谱法、电化学技术等分别测定FUen和FUenRu的体外抗癌活性、模拟生理环境下的油水分配系数以及其与DNA和胸苷酸合成酶的结合模式和结合系数等。再结合Amber、AutoDock、Gaussian等软件的量化模拟评价，深入分析揭示芳烃钌(Ⅱ)配合物靶向性的调控机制，进而筛选出疏水亲水平衡调节与靶向作用协同一致的目标前药配合物，并将芳烃钌(Ⅱ)配合物发展成为一种兼具抗癌活性与靶向性的药物运输工具。以上成果无疑将为抗癌药物与其运输载体的联合应用提供一套新颖的研究方案及有价值的调控机制。

“三脚钢琴凳”结构的二胺芳烃钌（Ⅱ）配合物是一种高效低毒的金属抗癌药物。基于疏水亲水平衡调节机制，在5-氟尿嘧啶的N1位引进疏水性的脂肪链和芳香环，再经希夫碱反应接入亲水性的二胺螯合配体，最后与同活性中心的芳烃钌前驱体反应，生成了一系列具有pH调控释放和光诱导激活潜能的异活性中心芳烃钌（Ⅱ）配合物。. 采用量化计算软件对所合成的物质进行结构分析与活性预测，重要的构效关系如下：①疏水亲水改性可提升药物对TS（胸苷酸合成酶）、DNA的结合能力，邻环效应对药物的靶向性识别有影响；②光照可同时激活两活性中心，含硝基的配合物可被更低能量的光激活；③双靶点芳烃钌（Ⅱ）配合物对DNA具有优良的靶向识别作用，通过pH调控释放和光诱导激活还可展现出对TS强烈的二次靶向识别。. 本项目的设计构思新颖，考虑了药物运输的疏水亲水平衡调节作用，结合了药物感应pH调控释放与光诱导激活原理，构建起5-氟尿嘧啶与芳烃钌（Ⅱ）双活性中心的协同调节机制，研究结果将拓宽人们对多靶点药物设计的认识，丰富异活性中心抗癌药物研究的内涵。