M2L2型水溶性金属-药物配合物的定向合成与抗肿瘤活性研究

The study of metal-drug complexes (MDCs) is one of the active areas of drug research, especially discrete MDCs. Base on the previous study, 3,5-bis(arylidene)-4-piperidone unit was evaluated as the effective centre of antitumor. In this study, some 3,5-bis(arylidene)-4-piperidone bridging schiff-base ligands will be designed and prepared by quaternization and schiff-base reaction, which are water-soluble. The directed coordination chemistry between these novel ligands and non-toxic metal ions, such as Pt2+, Zn2+, Fe2+ will be investigated to get discrete and water-soluble bimetallic macrocycle (M2L2-MDCs). New M2L2-MDCs will be fully characterized by NMR, MS, IR, X-ray single-crystal diffraction, powdered X-ray diffraction and elemental analysis. In vitro, new ligands and M2L2-MDCs will be evaluated against human carcinoma cell lines SW1990, MIA PaCa-2, PG-BE1, NCI-H460, and SK-BR-3 for antitumor activity by the CCK-8 method. In vivo, some ligands and M2L2-MDCs with better antitumor activity will be verified by building subcutaneous transplantation tumor nude mouse model for the most sensitive cell line. Some M2L2-MDCs candidate drugs will be screened because of their better cytotoxic and antitumor activities. In addition, the degradation regularity and the feasibility of the prepared M2L2-MDCs as materials for drug release will be discussed primarily. This study can effectively exploit the study area of MDCs as antitumor drugs and will provide theoretical and experiment basis for study of new antitumor drugs.

金属-药物配合物(MDCs，Metal-Drug Complexes)是药物研究的热点之一，特别是水溶性离散型MDCs极具研究价值。前期工作显示3,5-二芳亚甲基-4-哌啶酮衍生物具有抗肿瘤活性高、细胞毒性小等优点，本项目设计以3,5-二芳亚甲基-4-哌啶酮为“药效团”和桥联基团的双Schiff-base配体，通过季铵化改善水溶性，与M(II)（Pt2+、Zn2+、Fe2+等）定向组装水溶性M2L2-MDCs，通过NMR、MS、IR、X-Ray、PXRD等进行结构表征。体外采用CCK-8法，评价对SW1990、MIA PaCa-2、PG-BE、NCI-H460、SK-BR-3等抗肿瘤活性，体内建立最敏感细胞株的裸鼠皮下移植瘤模型，筛选出抗肿瘤活性好和细胞毒性小的候选药物；初步研究M2L2-MDCs在生理条件下的降解规律及5-FU的缓释性能，为新型抗肿瘤药物的开发提供理论和实验依据。

[背景]金属-药物配合物(MDCs，Metal-Drug Complexes)是药物研究的热点之一，特别是水溶性离散型MDCs极具研究价值。本项目选用抗肿瘤活性较好的新型3,5-二芳亚甲基-4-哌啶酮（3,5-Bis(Arylidene)-4-Piperidone，BAP）衍生物作为有机配体，与无毒的金属离子定向组装M2L2-MDCs，期望合成并筛选出抗肿瘤活性好和细胞毒性小的MDCs。前期工作和文献综述显示， BAP属于姜黄素类似物，可通过激活Caspase-3导致肿瘤细胞DNA断裂诱导细胞凋亡，也可诱导肿瘤细胞自噬，且细胞毒性小等优点。. [主要研究内容、重要结果、关键数据] . （1）本项目设计并合成以BAP为“药效团”的双Schiff-base配体75个，通过MTT法评价它们对HePG2、HeLa、K562、THP-1、A549、SGC7901等细胞系的抗肿瘤活性以及对人正常肝细胞LO2的细胞毒性，筛选出活性较好的BAP衍生物12个（部分IC50值小于3 μM）；. （2）进一步与M(II)（Zn2+、Fe2+离子）定向组装成24个M2L2-MDCs。体外采用MTT法，评价抗肿瘤活性，结果显示，部分MDCs的抗肿瘤活性较BAP有显著提高，且经季铵化修饰的BAP形成的MDCs水溶性显著提高，且抗肿瘤活性较好、细胞毒性较低。. （3）结合以上实验，筛选得到溶解性、抗肿瘤活性和稳定性较好的化合物Zn-L2-MDC，体内通过HepG2诱发的肝癌裸鼠移植瘤模型进一步证实了该化合物能有效抑制肝癌的细胞增值，并且对裸鼠无明显的毒性；. （4）初步研究Zn-L2-MDC在生理条件下对5-FU的载药性能，结果显示Zn-L2-MDC在模拟胃液和肠液中对5-FU的药物包封率达到35.5%，前12小时的药物缓释达到84.8%，12-72小时缓释较慢，在72小时左右载体释药量基本达到最大，显示了较好的缓释效果。. [科学意义]本项目利用3,5-二(芳亚甲基)-4-哌啶酮药效团桥联有机配体定向组装金属-药物配合物（M2L2-MDCs），并应用于体内和体外抗肿瘤药物的筛选，初步探索M2L2-MDCs在生理条件下的降解规律和药物缓释效果，为拓宽MDCs的应用范围和开发新型抗肿瘤药物提供理论和实验依据。