新型苯并咪唑类微管蛋白抑制剂的设计、合成及活性研究

发布时间：2018-03-12 07:03

本文选题：微管蛋白抑制剂　切入点：苯并咪唑　出处：《吉林大学》2014年硕士论文　论文类型：学位论文

【摘要】：微管是细胞骨架的基本组成部分，参与维持细胞形态、细胞内物质输送、信号传导、细胞分裂等重要过程。微管蛋白作为微管的基本单位，是抗肿瘤药物研究的重要靶点之一。目前有多种微管蛋白抑制剂作为一线抗肿瘤药物应用于临床，在延长患者生存时间和提高生活质量方面发挥重要作用。然而，现有的微管蛋白抑制剂普遍存在结构复杂难于合成、容易产生获得性耐药、毒副作用大等缺陷。因此，寻找作用机制新颖、高效低毒的小分子微管蛋白抑制剂已成为当今社会研究的热点之一。苯并咪唑类抗寄生虫药物广泛应用于临床，作用机制是与寄生虫的微管蛋白结合，对哺乳动物宿主几乎不产生影响。结构类似的苯并咪唑类化合物61却能够以新的作用模式与哺乳动物的微管蛋白结合。因此，本文将化合物61作为先导化合物，设计合成了一系列苯并咪唑类衍生物，期望得到化学结构简单、作用机制新颖、抗肿瘤活性显著的新型微管蛋白抑制剂。本文以5-氟-2-硝基苯胺和不同取代的苯酚为起始原料，，通过亲核取代、硝基还原、合环、氨化、水解、酰化等反应，共计合成26个目标化合物，经数据库检索结构均为首次报道。采用肿瘤细胞生长抑制实验对上述目标化合物的抗肿瘤活性进行筛选，结果显示，该类化合物对肿瘤细胞系NCI-H460、COLO205、K562、A431、HepG2、Hela和MDA-MB-435S均具有较好的抑制活性。其中化合物65、71、80和82的抗肿瘤活性与阳性药紫杉醇相当，活性最好的化合物80对肿瘤细胞系NCI-H460、COLO205、K562、A431、HepG2、Hela和MDA-MB-435S的IC50值分别为0.040、0.050、0.006、0.026、1.774、0.452和0.052μM（紫杉醇IC50值分别为0.010、0.003、0.004、0.007、0.990、0.410、0.009μM）。本文还以活性最好的化合物80为代表对苯并咪唑甲脲类化合物的作用机制进行了初步探讨。免疫荧光实验和肿瘤细胞周期阻滞实验结果显示，化合物80在浓度为0.100μM时即可抑制NCI-H460细胞纺锤体的形成，使其停滞于细胞分裂周期的G2/M期，并具有剂量依赖性。表明该类化合物能够通过影响肿瘤细胞微管蛋白正常功能的发挥，抑制纺锤体形成，影响肿瘤细胞正常周期的运转，诱导凋亡。计算机模拟模型显示，化合物80分别与β-微管蛋白Glu71形成一个氢键，与α-微管蛋白Arg2形成两个氢键、Asp251形成一个氢键，可能以全新的结合模式作用于微管蛋白αβ异二聚体相连的界面。初步的构效关系研究表明，苯并咪唑环C-2位反式酰胺上连接甲基氨基时，化合物活性显著提高，用乙基氨基、乙基、环丙基、苄基替代时活性均有所下降。苯并咪唑环C-5位上连接苯氧基时，化合物活性最好，用1-萘氧基、环己氧基及苄氧基替代时化合物活性均有一定程度的降低。苯并咪唑环C-5位上苯氧基的苯环间位连接取代基时化合物活性最好，其次是对位，取代基的种类对化合物活性影响不大。本文为研发化学结构简单、作用机制新颖、抗肿瘤作用显著的新型微管蛋白抑制剂提供了新的思路。
[Abstract]:Microtubules are the basic components of the cytoskeleton, involved in the maintenance of cell shape, intracellular material transport, signal transduction, cell division and other important process. As the basic unit of tubulin microtubules, is one of the most important targets for anticancer drug research. There are a variety of tubulin inhibitors as first-line anticancer drugs in clinical application, play an important role in prolonging the survival time of patients and improve the quality of life. However, tubulin inhibitors the current complex structure is difficult to synthesis, easy to produce drug resistance defects of high toxic side effects. Therefore, looking for the mechanism of novel small molecule tubulin inhibitors with high efficiency and low toxicity has become one of the research focus of today's society.
Benzimidazole antiparasitic drug widely used in clinical, the mechanism is combined with tubulin of mammalian host parasite, almost no impact. Benzimidazole compounds with similar structures 61 are able to tubulin role model and mammalian new combination. Therefore, this paper will compound 61 as a lead compound, a series of benzimidazole the derivatives were designed and synthesized, desired chemical mechanism has the advantages of simple structure, novel, novel tubulin inhibitors antitumor activity significantly.
In this paper, 5- fluorine -2- nitro aniline and different substituted phenols as starting materials by nucleophilic substitution, reduction of nitro group, cyclization, hydrolysis, amination, acylation reaction, total synthesis of 26 target compounds, the database structure are reported for the first time. By screening, growth inhibition of antitumor activity of the target compounds the tumor cells showed that NCI-H460, the compounds on tumor cell lines COLO205, K562, A431, HepG2, Hela and MDA-MB-435S have good inhibitory activity. The antitumor activity of compounds 65,71,80 and 82 with positive drug paclitaxel, 80 compounds the best activity of NCI-H460, COLO205 cell lines K562, A431, HepG2, Hela and MDA-MB-435S IC50 values were 0.040,0.050,0.006,0.026,1.774,0.452 and 0.052 M (paclitaxel IC50 = 0.010,0.003,0.004,0.007,0.990,0.410,0.009 M).
Based on the 80 best represented the activity mechanism of benzimidazole methyl urea compounds were discussed. Immunofluorescence assay and tumor cell cycle arrest results showed that 80 compounds at the concentration of NCI-H460 can inhibit the formation of spindle cells of 0.100 M, the arrest in G2/M cell cycle period. In a dose-dependent manner. Indicated that the compounds can affect tumor cells through tubulin function, inhibit the formation of spindle, the influence of tumor cell apoptosis induced by the normal cycle of operation. The computer simulation model shows that compound 80 and beta tubulin Glu71 forms a hydrogen bond, and alpha tubulin Arg2 form two a hydrogen bond, Asp251 forms a hydrogen bond, may be in the alpha beta tubulin dimer linked to ISO two new combination model of the role of the interface.
Preliminary study on structure-activity relationship showed that when connecting methyl amino benzimidazole ring C-2 trans amide compounds, activity was significantly increased with ethyl amino ethyl, cyclopropyl, benzyl substitution decreased. C-5 benzimidazole ring connected phenoxy compounds, the best activity, with 1- naphthyloxy reduce, cyclohexyloxy and benzyloxy substitution compounds to a certain extent. Benzimidazole ring position C-5 phenoxy benzene compounds between the best activity of substituent, followed by Para, types of the substituents has little influence on the activity of the compounds.
This paper provides a new way of thinking for the new microtubulin inhibitor with simple chemical structure, novel mechanism and a significant anti-tumor effect.

【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R914

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