基于两靶点设计的多核铂化合物的合成及其抗非小细胞肺癌多药耐药机制研究

Platinum drugs as the first-line drug for the treatment of non-small cell lung cancer (NSCLC) have the characteristics of effective treatment and inhibition of tumor effect. However, long term use of them will lead to severe multidrug resistance (MDR). At present, simple structural modification of mononuclear platinum complexes can not achieve the purpose of significantly reducing the MDR of NSCLC cells. Our previous study shows that polynuclear platinum complexes bind to DNA in a unique way. In addition, it is reported that polynuclear platinum complexes are combined with DNA by means of interlink crosslinking, so they exhibit significant effect of inhibiting tumor cell MDR. Based on this, with inhibiting tumor MDR of polynuclear platinum complexes as the starting point, a series of glycosylation thiosemicarbazone polynuclear platinum complexes will be synthesized in this study. They not only target the tumor cells through glucose transporter protein, but also target lysosomal by P-gp protein which will reduce the platinum drug efflux and inhibit cell MDR. Meanwhile, the researches about activity evaluation of anti-NSCLC cell, platinum drugs in cell and tissue targeting and the molecular mechanism of affecting DNA will be carried out. And then, we will optimize their structures and explore the mechanism of anti-tumor MDR from the perspective of dual-target and polynuclear platinum. The successful implementation of the project will provide theoretical and experimental basis for the development of a new type of platinum drugs with targeting, water solubility and inhibition of NSCLC cell MDR.

铂类药物作为治疗非小细胞肺癌(NSCLC)的一线药物，具有疗效确切，抑制肿瘤作用强等特点，但患者长期使用会产生严重的多药耐药(MDR)性。目前，仅对单核铂进行简单结构修饰无法达到显著降低NSCLC细胞MDR的目的。申请人前期研究发现，多核铂作用于DNA方式独特，且文献报道多核铂以链间交联等方式与DNA结合，能够抑制肿瘤细胞MDR。基于此，本项目以多核铂化合物抗肿瘤MDR为切入点，设计合成一类可借助葡萄糖转运体蛋白靶向肿瘤细胞，且可通过P-gp蛋白靶向溶酶体减少铂药外排，从而抑制肿瘤细胞MDR的糖基化缩氨基硫脲多核铂配合物。并通过对铂配合物进行抗NSCLC细胞活性评估，细胞和组织中的靶向探索以及其与DNA作用的分子机制研究，从两靶点和多核铂的角度优化铂药结构并探索其抗肿瘤MDR的作用机制。该项目的顺利实施将为开发靶向性强、水溶性好、可抑制NSCLC细胞MDR的新型铂药提供理论基础和实验依据。

铂类药物作为一线治疗癌症的有效药物之一，但铂类药物水溶性差，靶向性低，易引起肾毒性和耳毒性等严重不良反应，更严重的是随着治疗时间的延长，药物促使肿瘤细胞产生严重的多药耐药性，这导致90%的化疗失败。本项目设计合成一类可借助葡萄糖转运体蛋白靶向肿瘤细胞，且可通过Pgp蛋白靶向溶酶体减少铂药外排，从而抑制肿瘤细胞MDR的糖基化缩氨基硫脲核铂配合物。主要研究内容为：1）合成系列（Ⅰ）糖基化缩氨基硫脲配体及其铂配合物和系列（Ⅱ）氨基噻唑烷酮铂/铜配合物，完成对所合成配体及配合物的理化性质表征；2）研究通过MTT、单克隆形成实验评价新型糖基化缩氨基硫脲铂的抗肿瘤活性，使用CDocker计算并模拟了铂化合物分别与GLUT1和Pgp蛋白的结合能力，通过热迁移实验再次证明了化合物与两种蛋白的实际结合能力，蛋白化学抑制剂实验，凋亡观察，溶酶体靶向性观察等，探索糖基化缩氨基硫脲铂配合物抑制肿瘤细胞MDR的机制；3）研究通过5’-dGMP结合实验，质粒DNA核酸酶活性实验，紫外及荧光实验探索了糖基化缩氨基硫脲铂配合物和氨基噻唑酮铂/铜配合物抑制肿瘤的分子机制。该项目的顺利实施将为开发靶向性强、可抑制肿瘤细胞MDR的新型铂药提供理论基础和实验依据。