【作者】 赵卫权；

【导师】 崔承彬；

【作者基本信息】 中国人民解放军军事医学科学院 ， 药物化学， 2008， 博士

【摘要】 从天然产物中寻找抗肿瘤活性先导化合物对于开发新型抗癌药物具有积极的意义。本论文对药用植物细圆藤与三株放线菌抗生素抗性突变株的代谢产物及其抗肿瘤活性进行了研究。研究内容主要包括:细圆藤化学成分的提取分离、从无活性放线菌筛选获取活性突变株与三株放线菌活性突变株代谢产物的分离纯化、单体化合物的结构鉴定以及其抗肿瘤活性初步测试等。论文前半部分是关于细圆藤的化学成分及其抗肿瘤活性研究。细圆藤在《中华药海》中记录有治疗无名肿毒、疥疮肿毒、毒蛇咬伤、小儿惊风、破伤风等功效。鉴于前期研究中发现细圆藤枝叶提取物具有体外抗肿瘤活性而活性成分未见报道,本论文前半部分对细圆藤的化学成分进行了分离纯化、结构鉴定以及抗肿瘤活性的初步测试评价。结果从细圆藤枝叶中分离鉴定了五个化合物hopenone-B(1)、hopenol-B(2)、22-Hydroxyhopan-3-one(3)、erythrodiol 3-palmitate(4)和5β,24-cyclofriedelan-3-one(5)。其中,化合物4具有较强的抗肿瘤活性,在100μg/mL浓度下对K562细胞的抑制率为46.7%。本论文上述化合物1～5均为首次从该属中分离得到,其中化合物4为该植物抗肿瘤活性成分的首例报道,其抗肿瘤活性也属首次筛选发现。论文后半部分是有关从无活性放线菌野生菌株筛选获取活性突变株以及有关三株抗肿瘤活性突变株代谢产物及其抗肿瘤活性的研究内容。论文首先以一株没有抗肿瘤活性的海洋放线菌M15为出发菌株,采用抗生素抗性筛选技术即核糖体工程技术,开展了链霉素、新霉素、庆大霉素、利福平等抗生素抗性菌株筛选获取工作,从而获得了对不同抗生素产生抗性的突变株63株,再用突变株发酵产物采用MTT法筛选了抗肿瘤活性,结果得到有明显活性的突变株6株。继而,选择其中3株活性突变株,采用活性跟踪和薄层对照的方法,对与原始菌比较有差异的产物进行了跟踪分离,从中分离得到9个化合物,并分别鉴定为尿嘧啶(6)、染料木素(7)、对-甲酰胺苯甲酸(8)、2-吡咯酸(9)、邻二羟基苯甲酸(10)、过氧化麦角甾醇(11)、N-(2-羟基苯基)-乙酰胺(12)、大豆黄素(13)和1,9-二甲酯-吩嗪(14)。活性测试结果表明,化合物6、7、11、13和14对K562细胞有抑制作用,在100μg/mL浓度下的抑制率分别为41.4%、51.6%、28.1%、21.4%和25.2%。其中14为新天然产物,其抗肿瘤活性也属首次筛选发现。本论文上述研究结果表明,核糖体工程技术是一种有效的开拓药源微生物菌株资源的新方法。更多还原

【Abstract】 This dissertation describes the secondary metabolites and their anti-tumor activity from Pericampylus galucus and three actinomycetic antibiotic-resistent atrains.Studies include the isolation,identification and anti-tumor activities of the constituents from Pericampylus galucus,as well as the induction of a non-active actinomycetic strain,fermentation,screening of antibiotic-resistent Strains by MTT method using K562 cells and the isolation, identification and anti-tumor activities of the constituents from three actinomycetic antibiotic-resistent Strains.The front segment of dissertation is about isolation and identification of the constituents from Pericampylus galucus and their anti-tumor activities.The bronches and leaves of Pericampylus galucus has been used as a folk medicine in Yunnan province of China to cure tumor,itch,tetanus,eclampsia et al.The alcohol extract of its bronches and leaves with anticancer activity screened by MTT method showed anti-tumor activities.In order to elucidate the anti-tumor constituents,the chemical and biological investigation had been carried out.The results are fellow as:five triterpenoids 1-5 were isolated from Pericampylus glaucus and they were identified as Hopenone-B(1),hopenol-B(2),22-Hydroxyhopan-3-one (3),erythrodiol 3-palmitate(4)and 5β,24-cyclofriedelan-3-one(5);Among them,4 inhibited the proliferation of K562 cells with the inhibition rate of 47%at 100μg/mL; Compounds 1-5 were isolated from the genus Pericampylus for the first time,4 provides the first example of anti-tumor constituents of Pericampylus glaucus and its anti-tumor activity was also assayed for the first time.The rear segment of dissertation is about the method of a non-active Actinomycetic strain chosen as a original strain and induced the antibiotic resistant,isolation and identification of the constituents from three actinomycetic antibiotic-resistent Strains,preliminary evaluation for anti-tumor activities of pure compounds.A strain M15 shows no anti-tumor activities and was chosen as a original strain and then was induced the antibiotic(streptomycin,neomycin, gentamicin,Rifampicin)resistant i.e.ribosome engineering technology.63 strains of mutation were obtained,and their fermentation with anti-tumor activity screened by MTT method were examined using K562 cells.6 strains of 63 shows anti-tumor activities.3 of 6 strains having anti-tumor activities was to elucidate the anti-tumor constituents with the method of Guiding by bioassay of anti-tumor and comparing with original strain.The results are fellow as:9 compounds 6-14 were isolated and identified as uracil(6),Genistein(7),4 -(methyamino)-benzoic acid(8),1H-pyrrole-2-canboxylic acid(9),2,3-dihydrobenzoic acid (10),ergosterol peroxide(11),N-(2-hydroxyphenyl)-acetamide(12),Daidzein(13), 1,9-Dicarbomethoxy-phenazine(14).6,7,11,13 and 14 inhibited the proliferation of K562 cells with the inhibition rate of 41.4%,51.6%,28.1%,21.4%and 25.2 at 100μg/mL; Compounds 14 were isolated from the nature for the first time and its anti-tumor activity was also assayed for the first time.Above studies indicated that it is feasible to exploit new microbial resources for medicinal usage by applying the ribosome-engineering technology to non-active actinomycete in nature.更多还原