紫苏醇衍生物的设计、合成及抗癌活性研究

发布时间：2018-04-15 02:02

本文选题：紫苏醇 + 紫苏醇衍生物　；参考：《沈阳药科大学》2014年博士论文

【摘要】：紫苏醇是来源于植物的异戊二烯萜类挥发油,是为数不多的既有癌症预防又有癌症治疗作用的天然产物之一。紫苏醇具有良好的抗肿瘤作用,而且毒性小、不易产生耐药性。但紫苏醇在体内代谢迅速,口服给药生物利用度低,因而限制了其临床应用。本实验室对柠檬烯和榄香烯衍生物的研究发现,增加化合物的极性有利于抗肿瘤活性的提高。因此,本论文拟在紫苏醇结构中引入含氮基团,希望能获得抗肿瘤活性强,毒副作用小的新颖化合物。本文以紫苏醇为先导化合物,在保持其分子骨架的前提下,以紫苏醛为起始原料,首先合成了紫苏醇、7-乙基紫苏醇、7-甲基紫苏醇和紫苏酸甲酯四个母核,再以各种取代的胺基对其进行结构修饰,共设计合成了5个系列61个化合物,包括14个紫苏醇类衍生物(Al-A14)、11个7-乙基紫苏醇类衍生物(B1-B11)、10个7-甲基紫苏醇类衍生物(C1-C10)、18个紫苏酸甲酯类衍生物(Dl-D18)和8个紫苏胺类衍生物(E1-E8),其中59个未见文献报道。所有目标化合物的结构均经MS和1H-NMR确证。用MTT法对所合成的61个目标化合物进行了体外抗肿瘤活性筛选。以HeLa、MCF-7、HepG2、HCT116、A549、A375-S2、HT-1080、U937、HL-60和K562为测试细胞株,以5-氟尿嘧啶为阳性对照药,结果显示,所合成的大部分化合物对10种肿瘤细胞均表现出不同程度的抑制作用。说明含氮基团的引入确实有利于抗癌活性的提高,由此充分验证了本文的设计思想。对目标化合物构效关系进行初步探讨,结果如下：在紫苏醇以及7-甲基或乙基取代的紫苏醇含氮衍生物(A,B和C系列)中,取代基为伯胺的目标化合物的抗肿瘤活性强于仲胺；在紫苏酸甲酯的含氮衍生物(D系列)中,引入的不同的含氮取代基团对化合物的抗肿瘤活性并无显著的影响；紫苏醇结构中的羟基被氨基替换的紫苏胺类衍生物(E系列)中,引入金刚烷胺基的目标化合物E5抗肿瘤活性强于其他目标化合物。选取E5对其抗肿瘤作用机制进行了进一步研究。研究结果表明E5可以呈浓度依赖性、时间依赖性抑制A549细胞的增殖。用相差显微镜、荧光染色及流式细胞术检测表明,E5可引起A549细胞发生凋亡。免疫印迹分析结果证实了E5诱导A549细胞发生凋亡的作用,对法尼基转移酶有抑制作用,对Ras下游Raf/MEK/ERK信号通路有抑制作用。研究结果表明：E5通过抑制法尼基转移酶,从而抑制Raf/MEK/ERK信号通路而诱导细胞凋亡。
[Abstract]:Perilla alcohol is a volatile oil of isoprene terpenoids from plants. It is one of the few natural products which can prevent and cure cancer.Perilla alcohol has good anti-tumor effect, and it is not easy to produce drug resistance because of its small toxicity.However, the rapid metabolism of perilla alcohol in vivo and low bioavailability of oral administration limit its clinical application.Our laboratory studies on limonene and elemene derivatives show that increasing the polarity of the compounds is beneficial to the increase of anti-tumor activity.Therefore, in this paper, nitrogen-containing groups were introduced into the perilla alcohol structure in order to obtain novel compounds with strong anti-tumor activity and less toxic side effects.In this paper, under the premise of keeping the molecular skeleton of perilla alcohol as the leading compound, four parent nuclei of perilla alcohol 7-ethyl perilla alcohol 7-methyl perilla alcohol and methyl perilla acid were synthesized with perilla aldehyde as the starting material.Five series of 61 compounds were designed and synthesized by modifying them with various substituted amino groups.It consists of 14 perilla alcohols, 11 7-ethylperilla derivatives B1-B11, 10 7-methylperilla derivatives, C1-C10, 18 methyl perilla derivatives, Dl-D18) and 8 perilla derivatives, E1-E8, among which 59 have not been reported in the literature.The structures of all the target compounds were confirmed by MS and 1H-NMR.The anti-tumor activity of 61 target compounds was screened by MTT method in vitro.The HL-60 and K562 cell lines of HL-60 and K562 were tested with HCT116A549-A375-S2HT-1080 and 5-fluorouracil as positive control. The results showed that most of the synthesized compounds showed different inhibitory effects on 10 kinds of tumor cells.It shows that the introduction of nitrogen-containing groups is beneficial to the improvement of anticancer activity, which fully validates the design idea of this paper.The structure-activity relationship of the target compounds was preliminarily discussed. The results were as follows: the antitumor activity of the target compounds with primary amine was stronger than that of secondary amines in perilla alcohol and nitrogen-containing derivatives of 7-methyl-or ethyl-substituted perilla alcohol.In D series of nitrogen-containing derivatives of methyl perilla, the different nitrogen-containing substituents had no significant effect on the antitumor activity of the compounds.The anti-tumor activity of target compound E 5 was stronger than that of other target compounds.E5 was selected to further study the mechanism of its anti-tumor effect.The results showed that E5 could inhibit the proliferation of A549 cells in a concentration-dependent and time dependent manner.Phase contrast microscopy, fluorescence staining and flow cytometry showed that E5 could induce apoptosis of A549 cells.Western blot analysis confirmed that E5 induced apoptosis in A549 cells, inhibited farinotransferase and inhibited Raf/MEK/ERK signaling pathway downstream of Ras.The results showed that the cell apoptosis was induced by inhibiting the Raf/MEK/ERK signaling pathway by inhibiting farinotransferase.
【学位授予单位】：沈阳药科大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R914.5;R96

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