基于中药活性成分金属抗肿瘤配合物研究

In order to improve the serious side-effect and resistence of metal-based antituour agents, we adopt the following strategies: 1）The selectivity is increased by targeting design basing on estrogen receptor or focusing on G-quadruplex DNA; 2) By means of exerting the diversity and validity of traditional Chinese medicines active ingredients (TCMAI) and their synergetic effect on metal, the validity of antitumour TCMAI-metal complexes is enhanced; 3) By introducing other ligands, the systemic toxicity is reduced; 4) In virtue of multi-target and multi-machanism of TCMAI-metal complexes, the resistence is overcomed. By selecting the traditional Chinese medicine active ingredient oxoaporphine and oxoisoaporphine alkaloids as main ligands and adopting mentioned above strategies, we design and synthesize TCMAI-Pt/Ru complexes targeting G-quadruplex DNA or having estrogen receptor regulating activity. We carry out the screen on druggable target and the assay of in vitro antitumour activity; The transplant tumour validity evaluation of candidates on SCID mouse model, the PK-PD study on the combination of pharmacokinetics (PK) and pharmacodynamics (PD) and toxicology are performed. Basing on the human serum albumin, we construct the multi-targeting carrier of TCMAI-metal-based drug. By meansi of many analytical techniques, the action components, accumulation and distribution of TCMAI-metal complex in cells are detected. We investigate the effect of candidates on the tumour cell cycle, and study the apoptosis mechanism of tumour cells. By gene-chip technique, the relevant proteins and genes on tumour cells death are found. We validate the key targets and pathways with modern molecular biology methods. Finally, the new highly active, low toxic antitumour TCMAI-metal complexes with multi-target and multi-mechanism are obtained.

为改善金属抗肿瘤药物严重毒副作用和耐药性，拟针对雌激素受体或G-四链体DNA进行靶向设计以提高选择性；发挥中药活性成分(TCMAI)多样性和有效性及其与金属的协同作用,以提高其金属配合物的有效性；引入其它配体以降低系统毒性；发挥TCMAI－金属配合物的多靶点和多重作用机制以克服耐药性。选择中药活性成分氧化(异)阿朴菲为主要配体，采用上述策略，设计、合成雌激素受体调节或针对G-四链体DNA的TCMAI-铂、钌配合物。进行成药靶点和抗肿瘤活性筛选、裸鼠移植肿瘤有效性评价、PK-PD和毒理学研究。构建人血清白蛋白-TCMAI-金属药物多靶向载体，提高选择性和降低毒性。采用多种分析技术测定候选化合物在细胞中的作用成分、累积和分布。研究肿瘤细胞增殖周期和凋亡机制；利用基因芯片寻找肿瘤细胞死亡的相关基因和蛋白，用分子生物学方法验证关键作用靶点和通路。最终获得具有多靶点和多重作用机制的金属抗肿瘤配合物。

发挥中药活性成分(TCMAI)多样性和有效性及其与金属的协同作用，改善抗肿瘤金属药物严重毒副作用和耐药性；提高抗肿瘤先导金属化合物的成药性，提高先导金属化合物在体内的靶向性、药效、生物利用度和降低其毒副作用，是本项目的出发点和落脚点。本项目基于DNA、G4-DNA及相关酶、抗凋亡蛋白Mcl-1等靶标，提出和实现了中药活性成分与金属活性中心配位调控效应设计金属药物的思想。从广西民族药分离或合成了系列抗肿瘤、抗病毒新活性化合物120多个；发现了系列具有多靶点、多通路和多重作用机理的高活性抗肿瘤金属化合物90余个，获得先导化合物9个，完成1个先导化合物初步的药代动力学和毒理学研究。初步揭示了ACF7蛋白参与时空调控的结构机制；创新地提出了基于人血清白蛋白IB、IIA亚域和肿瘤细胞特性设计抗肿瘤金属前药的研究思想，获得有应用前景的金属前药4个；针对肿瘤组织的微环境和肿瘤的复杂性，构建了人血清白蛋白多药物递送体系多靶点协同联合治疗肿瘤模型3个。在Nat. Commun., J. Med. Chem., Chem. Commun., Org. Lett., Eur. J. Med. Chem., Mol. Pharm., J. Nat. Prod.等发表论文84篇，获授权中国发明专利42项。获广西科学技术奖特别贡献奖和广西自然科学奖二等奖各1项。主办承办国际国内会议10次；在国际学术会议做大会报告2次和邀请报告14次。培养国家“万人计划”领军人才和国家“百千万人才工程”人选各1名，广西“十百千人才工程”人选和广西特聘专家各1名。培养博士后、博士12名，硕士25名。本项目成果开拓了中药活性成分金属药物研究领域，为金属药物研究提供了新模式和新策略；为人血清白蛋白-金属药物复合物靶向治疗肿瘤，也为基于细胞迁移相关蛋白研发金属抑制剂提供了科学依据。