兰州大学机构库 >生命科学学院
γ-羟基炔酸酯类衍生物抗肿瘤及抗炎活性研究
Alternative TitleSynthesis of γ-hydroxy acetylenic esters derivatives and their evalution as anti-cancer and anti-inflammatory agents
来路皓
Thesis Advisor王锐
2017-05-08
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name博士
Keywordγ-羟基炔酸酯 靛红 抗肿瘤 凋亡 抗炎
Abstract

从远古以来人们就使用天然药物来治疗病痛。天然产物来源的药物具有多种多样的化学分子形式,其中聚炔酯类的结构是一类常见的具有活性的药物分子。广泛存在于各种各样的植物及海洋生物中。随着化学技术的发展和人们认识的进步,近几十年来,人们从传统药用植物或其它有活性的生物中分离出了上千种含炔类的活性成分。这些含有炔基的活性成分,表现出了丰富的生物活性,如抗肿瘤,抗炎,抗菌,抗病毒等。为了探讨炔酯类结构对药物活性的影响,在实验室此前的γ-羟基炔酸酯的不对称合成方法基础上,我们合成了一系列的化合物,把含有丙炔酸酯的基团引入到不同的具有药效活性的小分子基团上,并且检测了所得到的化合物的抗肿瘤以及抗炎的活性。我们的工作主要包括以下几个方面。<一>γ-炔酸酯类化合物的设计合成:(1)通过条件优化,在靛红(2,3-二酮二氢吲哚)3-位上引入丙炔酸甲酯基团,得到一系列的3-羟基氧化吲哚类似物,在对吲哚环上不同取代基进行替换同时,,亦对1位氨基的取代基也进行了筛选,(2)在此基础上,通过对靛红衍生的3-亚胺的加成得到一系列3-氨基氧化吲哚类似化合物;(3)此外,利用类似方法还制备了其他杂环衍生的γ-炔酸酯类化合物。<二>化合物的抗肿瘤活性评价。(1)首先利用Hela和MDA两种细胞系对所得化合物进行了肿瘤细胞增殖的抑制活性筛选。结合所测得的IC50对化合物进行了构效分析,并且根据构效关系进一步对化合物进行了基团改造。实验发现在靛红衍生的3-位羟基γ-炔酸酯化合物具有较好的抗肿瘤活性。对1-位进行基团替换即引入较大基团能显著改善化合物的抗肿瘤活性。其中化合物L-37表现出最优的抗肿瘤活性。(2)对具有代表性的化合物L-37,L-39的抗肿瘤机制进行了探讨。确定了合成的化合物作用是通过凋亡途径抑制肿瘤细胞生长的,并且呈时间和剂量依赖关系。随后的实验发现,该类化合物通过PI3K/AKT通路对凋亡蛋白caspase-9进行了激活,从而由caspase-3/9的通路诱导了肿瘤细胞的凋亡(3)另外根据罗丹明123和ROS含量检测实验发现,所合成化合物促进了肿瘤细胞内ROS的显著提高,表明所此类化合物物也引起了肿瘤细胞线粒体膜电势的改变,从而促使了细胞凋亡。因此,本论文所涉及的化合物通过多种途径引起肿瘤细胞凋亡,从而有效的抑制了肿瘤细胞增殖。<三>化合物的抗炎活性评价。鉴于此类化合物具有与抗炎成药吲哚美辛类似的结构特点,因此在进行抗肿瘤活性实验的基础上,对其抗炎活性进行了作用评价。(1)选取在抗肿瘤活性中表现良好的L-37,L-39,L-18等化合物,利用RAW264.7细胞进行了NO释放抑制作用的检测。结果发现该类化合物表现了出色的抗炎活性,1 μ  M浓度的L-37下即对LPS刺激RAW的NO释放抑制率达到50%;(2)酶学水平实验表明此类化合物对PGE2具有显著的抑制效果,并发现所合成的化合物对COX-1,COX-2,5-LOX酶的表达亦有一定的影响,这可能是此类化合物抗炎作用的另一作用途径。(3)小鼠抗炎实验结果表明化合物L-37具有和成药塞来昔布相当的体内抗炎作用,表明此类化合物在抗炎活性方面具有潜在的应用价值。我们合成的靛红炔酸酯类化合物,明显的表现出抗肿瘤和抗炎的双重活性,因炎症和肿瘤有着密不可分的关系,作为双重抑制剂,该类化合物有着巨大的开发潜力。

Other Abstract

Drugs from natural products have been chosen to treat diseases since antiquity.Polyacetylenic are a unique structure with biological functions as one of the wide variety drug candidates from natural products, widely existing in kinds of plants and marine organism. With the development of chemical technology and studies during last decades, thousands of active acetylenic metabolites have been isolated and identified from traditional medicine plants, which display abundant biological activities, such as anticancer, anti-inflammatory, antifungal, antimicrobial, antiviral. But the mechanism of these compounds is still not clear.In order to investigate the effort of acetylenic structure on the activity of drug, a series of compounds have been synthesized, based on the previous works for γ-hydroxyl acetylenic esters. These compounds, containing acetylenic ester groups into small medicine molecular, were detected kinds biological activity, contains antitumor and anti-inflammatory. The work mainly includes the following aspects.Design and synthesis of many kinds of compounds containing acetylenic ester. (1) Compounds obtained from isatin (1H-indole-2,3-dione), by replacement of substituent at the indole ring and addition of acetylenic ester in 3 keto. Synthesis methods have been optimized by attempt of kinds of catalyzer and experimental conditions. Not only introduction of the acetylenic ester at carbonyl group ,the replacement in 1 keto also plays important aspect in biological activity. (2) On the basis of previous work on the γ-hydroxy acetylenic esters in benzaldehyde, γ-amidogen acetylenic esters compounds were synthesized. (3) Acetylenic ester were introducted into kinds of heterocyclic group.Detected the anticancer activity of compounds.(1) Hela and MDA cell lines were chosen for the screening of the antitumor ability among obtained compounds. The structure activity analysis was carried out by the IC50 from activity test, and designing new compounds by replacement groups. Anticancer ability of isatin derivatives can increased dramatically by introduction acetylenic ester in carbonyl of 3-keto. Replacement of groups in 1-keto of insatin also can significantly improve the activity of compounds. The compound L-37 was the most potent in screening. (2) Compound L-37 and some others were selected for studying the mechanims of the antitumor. We confirmed the Hela cells were inhibited by apoptosis in a time and dose-dependent. In the subsequent experiments, compounds show the ability of inducing apoptosis through caspase-3/9 pathway, which may activated through the PI3K/AKT pathway.  (3)According to detection of Rhodamine 123 and ROS content, the compounds were promoted significantly increased ROS in tumor cells, showed that such compounds can induce mitochondrial membrane potential changes in tumor cells, which contributes to the cell apoptosis. Therefore, compounds involved in this study induced apoptosis of tumor cells through variety of ways, so as to effectively inhibit the proliferation of tumor cells.Detected the anti-inflammatory activity of compounds. As synthesised compounds have similar structure with indomethacin, the anti-inflammatory activity were detected based antitumor experiments. (1)L-37,L-39,L-18 and other compounds which showed good performance in antitumor exprements, were selected to detect NO release inhibition in RAW 264.7 cells. These compounds showed high anti-inflammatory ability as the anticancer activity, which can inhibite NO release in low level. Compound L-37 can inhibit NO release rate reached 50% in 1 μ M. (2) The enzyme experiment indicates that the compound has obvious inhibiting effect on PGE2, and found that the compounds showed same effect in COX-1, COX-2, 5-LOX expression.(3) L-37 listed similar anti-inflammatory effect in mice as celecoxib in the further animal experiments. The results show that the compounds have potential application in anti-inflammatory activitySynthesised compounds showed pronounced double activity of anti-tumor and anti-inflammatory. Due to inflammation and cancer are closely related, as a dual inhibitor, the compounds have great potential for development.

URL查看原文
Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/221814
Collection生命科学学院
Recommended Citation
GB/T 7714
来路皓. γ-羟基炔酸酯类衍生物抗肿瘤及抗炎活性研究[D]. 兰州. 兰州大学,2017.
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