16电子半三明治过渡金属配合物的合成及抗癌活性研究

【作者】 高洁；

【导师】 郭丽华；

【作者基本信息】 曲阜师范大学 ， 有机化学， 2022， 硕士

【摘要】 社会发展、环境污染和高强度工作使人们患癌的几率不断增加,因此发展抗癌药物具有举足轻重的作用。自顺铂问世以来,铂类金属抗癌药物在临床方面取得了优良的效果,但是因具有耐药性、高毒性等缺点限制了其临床应用。为了获得具有高抗癌活性、低毒副作用的抗癌药物,人们把目光聚集在其它金属抗癌配合物的设计与合成上。其中钌、铑、铱金属抗癌配合物因为化学性质稳定、具有低毒副作用而受到人们的广泛关注。α-酮基-β-二亚胺配体常用于合成高分子量聚乙烯和聚α-烯烃的高活性镍（II）引发剂,较少用于金属抗癌药物的合成。本文发展了一种更简便的方法合成了11个不同位阻的α-酮基-β-二亚胺配体,并且通过核磁共振光谱(¹H NMR、¹³C NMR),质谱和元素分析等方法对α-酮基-β-二亚胺配体进行了纯度、结构和组成的表征。并利用合成的小位阻α-酮基-β-二亚胺作为配体设计合成了5个半三明治结构铑配合物、1个半三明治结构铱配合物和1个半三明治结构钌配合物,对金属配合物光物理性质、溶液稳定性、DNA结合、亲脂性研究和体外抗癌活性进行了初步检测,选择生物活性较好的金属配合物进行细胞成像、细胞凋亡和细胞周期分析,具体如下:1、通过一步反应合成了一系列α-酮基-β-二亚胺配体。这些配体可以通过间氯过氧苯甲酸（m-CPBA）做为氧化剂一步合成,而不需要之前文献报道的三步反应。在此基础上,成功合成了16电子（不带离去基团Cl^-）的半三明治钌配合物。2、以往人们对于18电子半三明治铂族（Rh、Ru、Ir和Os）金属配合物的结构、抗癌活性和作用机制进行了广泛的研究,但16电子（不带离去基团Cl^-）半三明治金属配合物的合成和生物学评价却很少涉及。本文报道了以m-CPBA为氧化剂,通过重排配位反应合成了一组结构相关的16电子“钢琴凳”（piano-stool）铑、铱和钌配合物。通过核磁共振波谱、质谱、单晶X射线晶体学和元素分析等方法进行了表征。通过¹H NMR分析表明配合物在水相中具有良好的稳定性。尽管没有观察到配合物与碱基（9-Et G和9-Me A）的反应,但代表性的Rh5与CT-DNA之间表现出一定的相互作用。合成的16电子配合物对A549和He La癌细胞表现出良好的抗癌活性,IC₅₀值在7.1-32.3μM范围内,与顺铂相当,甚至优于顺铂。此外,在构效关系上,金属中心由铱（III）和钌（II）转变为铑（III）可增强配合物的细胞毒性。利用流式细胞术对其作用机制进行研究,发现配合物Rh5的细胞毒性与细胞周期的干扰和诱导细胞凋亡有关。利用共聚焦显微镜发现,配合物Rh5通过能量依赖途径进入A549细胞,并主要在溶酶体中积累,进而破坏溶酶体的完整性。更多还原

【Abstract】 With the development of society,environmental pollution and high-intensity work,the probability of people suffering from cancer is gradually increasing,so the development of anti-cancer drugs plays an important role.Since the advent of cisplatin,platinum-metal anticancer drugs have achieved excellent clinical effects,but their application in clinical treatment is limited by drug resistance and high toxicity.In order to obtain anticancer drugs with high anticancer properties and low side effects,people focus on the design and synthesis of other metal anticancer complexes.Among them,ruthenium,rhodium and iridium anticancer complexes have attracted extensive attention due to their stable chemical properties and low side effects.α-ketol-β-diimine ligands are commonly used as nickel（II）initiators for the synthesis of high molecular weight polyethylene and polyα-olefin,but rarely used in the synthesis of metal anticancer drugs.Therefore,the anticancer potential of metal complexes synthesized fromα-ketol-β-diimine ligands is great.In this paper,11α-ketonyl-β-diimine ligands with different steric hinds were synthesized in a simpler way than the previous methods.The purity,structure and composition ofα-ketonyl-β-diimine ligands were characterized by ¹H NMR,¹³C NMR,mass spectrometry and elemental analysis.Five half-sandwich rhodium complexes,one half-sandwich iridium complex and one half-sandwich ruthenium complex were synthesized by the synthesis of small hindranceα-ketol-β-diimine as ligands.They were characterized by¹H NMR,¹³C NMR,mass spectrometry and elemental analysis.The photophysical properties,solution stability,DNA binding,lipophile studies and preliminary detection of in vitro anticancer activity of metal complexes were investigated.Metal complexes with better biological activity were selected for cell imaging,cell apoptosis and cell cycle detection,as follows:1.A series ofα-ketol-β-diimine ligands were synthesized in one step.These ligands can be synthesized using m-chlorobenzoic acid（m-CPBA）as an oxidant in one step instead of the previous three steps.On this basis,16-electron（without the leaving group Cl^-）half-sandwich ruthenium complex was successfully synthesized.This novel complex offers great potential for metal anticancer drugs.2.The 18-electron half-sandwich platinum group（Rh,Ru,Ir and Os）metal anticancer complexes have been largely reported due to their easy tunable structure,high potency toward cancer cells and specific mechanism of actions.However,the16-electron（without the leaving group Cl^-）half-sandwich complexes and their biological evaluation are rarely investigated.With an easy access to the requiredα-keto-β-diimine ligands using m-CPBA as oxidant,we herein reported the synthesis and characterization of a panel of structurally related 16-electron piano-stool rhodium,iridium and ruthenium complexes through the rearranged coordination reaction.These complexes have been well-established by NMR spectroscopy,mass spectrometry,single-crystal X-ray crystallography and elemental analysis.Each of these complexes was stable and exhibited fluorescence in solution.Although no reaction with nucleobase（9-Et G and 9-Me A）was observed,the representative Rh5 showed affinity toward CT-DNA.These 16-electron complexes showed promising activity toward A549 and He La cancer cells with the values of IC₅₀in the range of 7.1-32.3μM,which was comparable to or even better than cisplatin.Further,The structure-activity relationships were observed that the change of the metal center from iridium（III）and ruthenium（II）to rhodium（III）could enhance the cytotoxicity of these unsaturated complexes.The investigation of mechanism of actions（Mo As）using flow cytometry displayed that the cytotoxicity of the complex Rh5 was associated with the perturbation of the cell cycle and the induction of cell apoptosis.Moreover,microscopic analysis by confocal microscopy suggested that the representative16-electron complex Rh5 entered A549 cells via energy-dependent pathway and predominantly accumulated in lysosomes,thus resulting in the disruption of lysosomal integrity.更多还原