荧光嘌呤核苷类似物合成及性质研究

发布时间：2019-06-12 06:34

【摘要】：目的:核苷是核酸(DNA或RNA)的基本组成部分。核苷衍生物广泛应用于药物研发、分子探针设计、癌症机理探讨等研究领域,同时也是临床常用的抗病毒、抗肿瘤等抗代谢类药物。核苷分子的碱基和糖基均可以进行化学修饰。碱基的修饰位点众多,嘌呤核苷的C-8位修饰是合成核苷衍生物重要途径,也是获得荧光核苷类似物的有效方法。目前,在2-氨基脱氧腺苷和异鸟苷的8位进行修饰的研究较少,本研究拟在2-氨基2'-脱氧腺苷和2'-脱氧异鸟苷的8位进行修饰,以期获取具有抗肿瘤活性以及能用作荧光探针的的新嘌呤核苷。方法:1化合物的制备卤代嘌呤核苷是碱基修饰的重要中间体。首先,将2-氨基-2'-脱氧腺苷溶于冰醋酸中,加入醋酸钠,滴加适量溴素,合成8-溴-2-氨基-2'-脱氧腺苷,接着,将其用于Sonogashira反应,溶于DMF溶液,加入Pd(PPh3)2Cl2/Cu I(1:2),三乙胺(3 eq)及相应取代炔烃,微波加热,合成一系列新的8位炔基取代的2-氨基-2'-脱氧腺苷衍生物;同样方法,用8-溴腺苷为前体,也合成出一系列8-取代腺苷衍生物。新的8位炔基取代的2-氨基-2'-脱氧腺苷衍生物在醋酸和Na NO2作用下通过生成重氮盐中间体,嘌呤环2位氨基转化为羰基,制备出一系列新的8位被炔基取代的脱氧异鸟苷2荧光性质研究荧光分光光度计测量化合物的荧光光谱、荧光量子效率及溶剂极性(二氧六环、甲醇和水)和溶液p H(1.5-12)值对化合物荧光光谱的影响。3化合物p Ka值的测定紫外分光光度滴定法测定脱氧异鸟苷衍生物的电离常数(p Ka);4抗肿瘤活性检测CCK-8法研究2-氨基-2'-脱氧腺苷、异鸟苷、腺苷衍生物对MM.1S人多发性骨髓瘤细胞生长的抑制作用,Graphpad prism 5统计实验结果,根据细胞存活率计算化合物的GI50。结果:8-溴-2-氨基-2'-脱氧腺苷的收率为87%;腺苷及2-氨基-2'-脱氧腺苷取代反应的产率为76-95%;2'-脱氧异鸟苷衍生物的收率为52-70%;所有化合物通过1HNMR和13C-NMR以及高分辨质谱分析,确定结构。苯炔衍生物取代的2-氨基-2'-脱氧腺苷和2'-脱氧异鸟苷有明显荧光,2'-脱氧异鸟苷衍生物的荧光强度对环境敏感,在二氧六环和水中呈现出不同的激发波长和发射波长以及不同的荧光强度,同时,2'-脱氧异鸟苷对p H敏感,可随着p H的增加而减弱;与2'-脱氧异鸟苷相比,8位炔基化的异鸟苷衍生物的p Ka值都发生显著变化;抗肿瘤活性检测发现苯炔衍生物取代的2-氨基-2'-脱氧腺苷和2'-脱氧异鸟苷化合物存在抑制MM.1S增殖的现象;测定化合物2a、2c、4、4a、4b、4c的GI50分别为24.28、21.11、7.87、13.78、10.08、7.10。结论:在微波辅助下,高收率的合成出一系列新的8位炔基取代的2-氨基-2'-脱氧腺苷和脱氧异鸟苷。8-位被芳香炔基取代的2-氨基-2'-脱氧腺苷、2'-脱氧异鸟苷可产生明显的荧光。其中,8位芳炔基化的2'-脱氧异鸟苷荧光分子对微环境敏感,具有作为荧光探针的前景。部分8-位芳炔基衍生物取代的腺苷、2'-脱氧异鸟苷具有抗肿瘤活性。
[Abstract]:Purpose: The nuclear antigen is a basic component of a nucleic acid (DNA or RNA). The nuclear antigen derivative is widely used in the fields of drug development, molecular probe design, cancer mechanism and the like, and is also an anti-metabolic drug commonly used in clinic, such as anti-virus, anti-tumor and the like. The base and the glycosyl of the nuclear antigen can be chemically modified. The modification site of the base has many modification sites, and the C-8-position modification of the methotrexate core is an important way to synthesize the nuclear antigen derivative, and is also an effective method for obtaining the fluorescent nuclear antigen analog. At present, the 8-bit modification of the 2-amino-2 '-deoxyadeno-and 2'-deoxy-avirences is less, and the study is intended to be modified at the 8-bit of the 2-amino-2 '-deoxyadeno-and 2'-deoxy-iopterin, with a view to obtaining a new methotrexate core with anti-tumor activity and which can be used as a fluorescent probe. Method: The preparation of a compound is an important intermediate of base modification. firstly, dissolving 2-amino-2 '-deoxyadenine in glacial acetic acid, adding sodium acetate, dripping an appropriate amount of bromine, and synthesizing 8-bromo-2-amino-2'-deoxyadenine, then, reacting with Sonogashira, dissolving in DMF solution, adding Pd (PPh3) 2Cl2/ Cu I (1:2), Triethylamine (3 eq) and corresponding substituted acetylenic hydrocarbon, microwave heating, synthesis of a series of new 8-bit alkynyl-substituted 2-amino-2 '-ethanal derivatives; the same method, with 8-bromothymol as a precursor, A series of 8-substituted adenine derivatives are also synthesized. The novel 8-position alkynyl-substituted 2-amino-2 '-deoxyadenine derivative is converted into a diazonium salt intermediate by the generation of a diazonium salt intermediate under the action of acetic acid and Na NO2, The fluorescence spectrum, the fluorescence quantum efficiency and the solvent polarity (dioxane, The influence of the value of methanol and water) and solution p h (1.5 -12) on the fluorescence spectrum of the compound. The inhibitory effect of the adenoid derivative on the growth of MM. 1S human multiple myeloma cells, and the results of the statistical experiment of Graphpad Prism 5, the GI50 of the compound were calculated according to the cell survival rate. The results showed that the yield of 8-bromo-2-amino-2 '-deoxyadenine was 87%. The yield of the adeno-and 2-amino-2 '-deoxyadeno-substituted reaction is 76-95%; the yield of the 2'-deoxy-heteroornithine derivative is 52-70%; all the compounds are analyzed by 1HNMR and 13C-NMR and high-resolution mass spectrum to determine the structure. the 2-amino-2 '-deoxyadeno-1 and 2'-deoxy-isoornithine-substituted 2-amino-2 '-deoxyadeno-1 and 2'-deoxy-isophorone derivatives have obvious fluorescence, the fluorescence intensity of the 2 '-deoxy-isoornithine derivative is sensitive to the environment, different excitation wavelengths and emission wavelengths and different fluorescence intensities are present in the dioxane and the water, At the same time,2 '-deoxy-isoornithine is sensitive to p H, and can be weakened with the increase of p H; compared with 2'-deoxy-bird's, the p-Ka value of the 8-position alkynylin derivative has a significant change; The anti-tumor activity test showed that the 2-amino-2 '-deoxyadeno-1 and 2'-deoxy-I-2 '-deoxy-I-1 compounds substituted by the benzalkynes inhibited the proliferation of MM.1S, and the GI50 of the compounds 2a, 2c,4, 4a, 4b, and 4c was 24.28, 21.11, 7.87, 13.78, 10.08, 7.10. Conclusion: Under the aid of microwave, A series of new 8-position alkynyl-substituted 2-amino-2 '-deoxyadeno-1 and deoxy-isoornithine are synthesized in a high yield. The 8-position is substituted with an aromatic alkynyl group of 2-amino-2'-pentadenoid, and the 2 '-deoxy heteroornithine can produce significant fluorescence. In which,8-position arginylation of 2 '-deoxy-isophorone fluorescent molecule is sensitive to the micro-environment and has the prospect as a fluorescent probe.
【学位授予单位】：成都医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R914

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