5,7-二甲氧基-1,4-萘醌衍生物的合成及抗细胞增殖活性机制研究

发布时间：2018-06-16 07:48

本文选题：拉帕醇 + 1　；参考：《兰州大学》2017年硕士论文

【摘要】：与正常细胞相比,肿瘤细胞具有较高的活性氧(ROS)水平和受损的抗氧化防御系统,因此,促ROS生成的药物可以通过加剧细胞的氧化应激状态来杀死癌细胞。更重要的是,由于正常细胞具有强大的抗氧化防御系统,所以可以抵御外来的ROS应激损伤。所以,促ROS生成的肿瘤药物,可以降低药物治疗的副作用。研究表明:1,4-萘醌类化合物可以增加细胞的ROS水平,进而杀死肿瘤细胞。本论文以拉帕醇为先导化合物,设计合成了一系列5,7-二甲氧基-1,4-萘醌衍1.目标化合物的设计:在文献调研的基础上,我们以2-羟基-1,4-萘醌为母核,将甲氧基引入芳环旨在增加醌环的亲电性和降低氧化还原电位;通过改变侧链长度和空间结构,旨在增加化合物的脂溶性和研究碳链的长度以及体积对抗细胞增值活性的影响;将相同的策略应用于2-甲氧基-1,4-萘醌和2-氯-1,4-萘醌系列。最后,我们利用成药5规则对设计的目标化合物进行初筛,发现基本满足成药5规则的要求。2.目标化合物的合成:根据目标化合物设计合成路线,对关键反应步骤:傅克酰基化反应和2-氯-1,4-萘醌系列合成的反应条件进行优化,最终获得最优的合成路线。3.目标化合物抗细胞增殖活性机制研究:通过MTT方法检测目标化合物对于六种肿瘤细胞A549(人非小细胞肺癌)、Hela(人宫颈癌细胞)、Hep-G2(人类肝癌细胞)、NCI-H460(人大细胞肺癌细胞)、HL60(急性白血病细胞)、K562(人慢性骨髓性白血病细胞)和两种人正常细胞WI-38(人胚胎肺成纤维细胞)、HEK 293(人类胚胎肾细胞)的抗细胞增殖活性。其中,化合物9e对HL60细胞的半抑制浓度IC50为3.80μM。相对于正常细胞WI-38细胞,其选择性指数为10.7,说明化合物在杀死癌细胞的同时,几乎不影响正常细胞。然后我们发现化合物9e可以诱导HL-60细胞凋亡,阻滞细胞周期在G2/M期,以及诱导ROS水平升高。加入抗氧化剂N-乙酰半胱氨酸(NAC),可以抑制凋亡,周期阻滞和ROS生成,表明ROS参与杀死细胞的过程。4.构效关系分析:随着烷基侧链变长,化合物7a-7i的抗增殖活性逐渐增加。具有环己烷或取代苯基团的化合物7j-7n表现出更好的抗增殖活性。化合物9e具有最好的抗增殖活性,表明C-2位置氯取代对其抗增殖活性非常有利。
[Abstract]:Compared with normal cells, tumor cells have higher reactive oxygen species (Ros) level and damaged antioxidant defense system. Therefore, Ros-promoting drugs can kill cancer cells by exacerbating oxidative stress. More importantly, normal cells have a strong antioxidant defense system, so they can resist external Ros stress damage. Therefore, tumor drugs that promote Ros production can reduce the side effects of drug therapy. It is shown that the Ros level of the cells can be increased by 1: 1 and 4-naphthoquinone compounds, which can kill the tumor cells. In this paper, a series of 5-dimethoxy-1-methoxy 4-naphthoquinone derivatives were designed and synthesized by using lappa alcohol as a leading compound. Design of target compounds: on the basis of literature review, we introduced methoxy into aromatic ring to increase the electrophilicity of quinone ring and reduce the redox potential by changing the length and spatial structure of the side chain. The aim of this study was to increase the liposolubility of the compounds and to study the effects of the length and volume of the carbon chain on the cell proliferation activity. The same strategy was applied to the series of 2-methoxy -1o 4-naphthoquinone and 2-chloro-1-butadiene 4-naphthoquinone. Finally, we use the 5 rule of patent medicine to screen the designed target compound, and find that it basically meets the requirement of the 5 rule of patent medicine. 2. Synthesis of target compounds: according to the design of synthesis route of the target compound, the key reaction steps: Fourier acylation reaction and the reaction conditions of 2-chloro-1n 4-naphthoquinone series synthesis were optimized, and the optimal synthesis route was obtained. Study on the mechanism of anti-proliferation activity of the target compound: MTT assay was used to detect the effect of the target compound on six kinds of tumor cells, A549 (human non-small cell lung cancer) Hela (human cervical cancer cell line Hep-G2) (human hepatocellular carcinoma cell line NCI-H460) Anti-proliferative activities of K562 (human chronic myeloid leukemia cell) and WI-38 (human embryonic lung fibroblast) and two human normal cells (human embryonic kidney cells). The IC50 of compound 9e on HL60 cells was 3.80 渭 M. The selectivity index of WI-38 cells was 10.7 compared with that of normal WI-38 cells, indicating that the compounds killed cancer cells and almost did not affect normal cells at the same time. Then we found that compound 9e could induce HL-60 cell apoptosis, block cell cycle at G _ 2 / M phase, and increase Ros level. The addition of the antioxidant N-acetylcysteine can inhibit apoptosis, cycle arrest and Ros production, suggesting that Ros is involved in the cell killing process. Structure-activity relationship analysis: as the alkyl side chain became longer, the antiproliferative activity of compound 7a-7i increased gradually. The compound 7j-7n with cyclohexane or substituted phenyl group showed better antiproliferative activity. Compound 9e has the best antiproliferative activity, indicating that the substitution of C-2 position chlorine is very beneficial to its anti-proliferation activity.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R914;R96

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