茚并吡唑类衍生物的合成与抗肿瘤细胞增殖活性

发布时间：2018-01-31 07:43

本文关键词： 优势结构茚并吡唑微管蛋白微管蛋白抑制剂抗肿瘤　出处：《山东大学》2017年硕士论文　论文类型：学位论文

【摘要】：优势结构是指能与多种生物受体进行有力结合或者作用的分子结构,具有"类药"、"类先导化合物"等特性。茚并吡唑是三环杂环结构,由苯环,中心5元环和吡唑环组成。作为"优势结构",茚并吡唑近年来已经广泛用于具有多种靶点的抗肿瘤药物设计,如作为检查点激酶-1,表皮生长因子受体,血管内皮生长因子,血小板衍生生长因子受体,细胞周期蛋白依赖性激酶和微管蛋白聚合的抑制剂等。在前期研究中,我们实验室发现了一个新型茚并吡唑类衍生物LL-01。LL-01作用于微管蛋白秋水仙碱结合位点,能够抑制微管蛋白的聚合,在体外对多种肿瘤细胞,包括多药耐药肿瘤细胞具有非常好的抑制活性。LL-01能使肿瘤细胞有丝分裂停滞于G2/M期,能诱导细胞凋亡,在体内具有良好的抗肿瘤活性。为进一步探讨该类化合物的构效关系,发现了活性更高的茚并吡唑衍生物,本研究拟在茚并吡唑的6位、7位进行结构修饰,并在3位苯胺取代基上引入乙胺基、乙氧基、氰基、酯基和酰胺基等,设计合成了 15个茚并吡唑类衍生物。以5,6-二甲氧基-1-茚酮为起始原料,经脱甲基反应后,乙基选择性地5位酚羟基反应,6位酚羟基经TBS保护,合成茚酮中间体,再与异硫氰酸酯衍生物亲核加成,经水合肼环合后,N原子甲基化,脱去6位羟基保护后,与氯乙酸甲酯经威廉姆逊成醚反应成醚,再进行氨解,制备了 10个化合物(A1-A10)。用MOM选择性保护5,6-二羟基-1-茚酮的5位酚羟基,6位羟基与3-氯-1-丙醇反应成醚,引入3-羟基丙基,再用MOM保护其伯羟基,制备茚酮中间体。与异硫氰酸酯衍生物经亲核加成,水合肼环合后,N原子甲基化,用樟脑磺酸脱去MOM保护基,合成了 6位酚羟基取代的茚并吡唑衍生物B1、B2和B3。将B1、B2 7位侧链上的伯羟基用TBS保护后,6位酚羟基与对甲氧基氯苄经威廉姆逊成醚,脱去TBS保护基后,合成了茚并吡唑衍生物B4和B5。对合成的化合物经1HNMR,13CNMR,ESI-Ms等进行了结构确证,并且进行了抗肿瘤细胞(K562,MCF-7,A549)增殖活性测试。初步地抗肿瘤活性筛选实验表明,大多数化合物对该三种肿瘤细胞的抑制活性显著优于ABT-751。在3-苯胺基的间位引入酯基,化合物A1的活性优于先导化合物LL-01。在化合物A1的7位侧链酰胺的氮原子上引入甲基或乙基,导致活性降低,而引入羟基,化合物A2的活性与LL-01相当。在3-苯胺基的间位引入氰基、甲酰胺基,活性降低。在茚并吡唑的6位引入对甲氧基苄氧基或羟基,化合物的活性大幅降低。通过本研究,发现了比LL-01活性更高的茚并吡唑衍生物,进一步的抗肿瘤药理学研究有待进行。
[Abstract]:The dominant structure refers to the molecular structure which can bind or interact with various biological receptors, and has the characteristics of "medicine-like" and "lead-like compounds". Ninopyrazole is a tricyclic heterocyclic structure consisting of benzene ring. As a "dominant structure", indenopyrazole has been widely used in the design of anti-tumor drugs with multiple targets, such as kinase-1 as a checkpoint, epidermal growth factor receptor. Vascular endothelial growth factor, platelet-derived growth factor receptor, cyclin dependent kinase and inhibitor of tubulin polymerization. We have found a novel indenopyrazole derivative LL-01.LL-01 acting on the tubulin colchicine binding site, which can inhibit tubulin polymerization in vitro for a variety of tumor cells. Including multidrug resistant tumor cells with very good inhibitory activity. LL-01 can make tumor cells mitosis arrest at G 2 / M phase, can induce cell apoptosis. In order to further study the structure-activity relationship of these compounds, a more active indenopyrazole derivative was found. In this study, the structure of ninopyrazole was modified at the 6 ~ 7 position. Fifteen derivatives of indenopyrazole were designed and synthesized by introducing ethylamino, ethoxy, cyanyl, ester and amideyl groups on the 3-position substituted aniline group. After demethylation reaction, ethyl-5-phenolic hydroxyl group was selectively reacted with 6-phenolic hydroxyl group, protected by TBS, the intermediate of indenone was synthesized, then nucleophilic addition of indenone with isothiocyanate derivative was carried out, and cyclized by hydrazine hydrate. The N atom was methylated, 6 hydroxyl groups were removed and then reacted with methyl chloroacetate to form ethers by Williamson and then ammoniated. Ten compounds, A1-A10, were prepared and reacted with 3-chloro-1-propanol to form ethers, which were selectively protected by MOM in the presence of 5-phenolic hydroxyl group and 6-position phenolic hydroxyl group. Indanone intermediate was prepared by introducing 3-hydroxypropyl group and protecting its primary hydroxyl group with MOM. The N atom methylated by cyclization of hydrazine hydrate was synthesized by nucleophilic addition with isothiocyanate derivative. The six phenolic hydroxyl substituted indenopyrazole derivatives B _ 1C _ 2 and B _ 3 were synthesized by removing MOM protection group from camphor sulfonic acid. The primary hydroxyl groups on B _ 1 / B _ 2 _ 7 side chain were protected by TBS. The 6-position phenolic hydroxyl group and p-methoxybenzyl chloride were synthesized by the synthesis of indolopyrazole derivatives B _ 4 and B _ 5. the synthesized compounds were synthesized by 1HNMR-13CNMR after the removal of the TBS protection group from the 6-position phenolic hydroxyl group and p-methoxybenzyl chloride (p-methoxybenzyl). ESI-Ms et al were used to confirm the structure and to test the proliferative activity of K562MCF-7A549. The results showed that the anti-tumor activity was screened. The inhibitory activity of most compounds on these three tumor cells was significantly better than that on ABT-751.Ester groups were introduced into the 3-aniline group. The activity of compound A1 is superior to that of lead compound LL-01.Introduction of methyl or ethyl to the nitrogen atom of the 7-position side chain amide of compound A1 leads to the decrease of activity and the introduction of hydroxyl. The activity of compound A2 was similar to that of LL-01. The activity of compound A2 was decreased by introducing cyanide and formamide at the m-site of 3-aniline group, and p-methoxy benzoxy group or hydroxyl group at the sixth position of indenopyrazole. The activity of the compounds was significantly reduced. In this study, ninhydropyrazole derivatives with higher activity than LL-01 were found, and further antitumor pharmacological studies were needed.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R914;R96

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