不对称阳离子表面活性剂的合成及其复配研究

发布时间：2018-04-21 21:51

本文选题：不对称阳离子表面活性剂 + 阴-阳离子表面活性剂复配　；参考：《大连理工大学》2015年硕士论文

【摘要】：近年来,基因治疗的研究一直是医学界最重要的领域之一,其对于肿瘤、免疫缺陷症等疾病已经用于临床；但是高毒性和相对较低的效率严重影响了其发展。相对于病毒载体的低安全性,非病毒载体的低免疫反应、低毒和易制备等优点使其成为研究热点。本论文设计合成了6种疏水链不对称的阳离子表面活性剂GHCC12-14、GHCC12-16、 GHCC14-16、GHBC12-14、GHBC12-16和GHBC14-16。通过1H-1H相关谱确定了长链异氰酸酯连接在N,N-二甲基氨基-1,2-丙二醇的端位羟基上；所有的中间体和目标产物都通过MS、1H NMR和1H-1H gCOSY表征,结构正确。考察了所合成的不对称阳离子表面活性剂的γcmc、cmc、HLB值和临界排列参数P等性质。发现随着碳链长度的增加表面张力和cmc增大,其中碳链长度为12-14的表面活性剂的γcmc和cmc都较小,具有较好的表面活性。将6种对称型的阳离子表面活性剂和六种不对称型的阳离子表面活性剂均与阴离子表面活性剂(SDBS、SDS和SAS)进行复合制备囊泡,对其进行外貌形态、粒径、Zeta电位、电导率和透射电子显微镜TEM表征,发现阴-阳离子表面活性剂复合体系不论是富阳离子区还是富阴离子区均能形成囊泡。除X1=0.5时会出现沉淀或聚沉,粒径基本都分布200～400 nm之间符合囊泡的范围；Zeta电位在富阴离子区小于-30 mV,在富阳离子区大于30 mV,说明混合溶液自发形成的囊泡稳定性较好。通过Masaniko几何模型和Israelachvili理论对囊泡的形成机理进行了阐述。将6种疏水链不对称型阳离子表面活性剂进行了生物学实验研究。TEM表明,除GHBC14-16以外,不对称阳离子类脂GHCn-m能够较好地形成脂质体；不对称阳离子脂质体与DNA复合后的粒径和Zeta电位的研究表明,其粒径大多在300-400 nm之间,符合细胞转染的要求；琼脂糖凝胶电泳实验结果显示,不对称阳离子脂质体GHCn-m与DNA的比例大于4：1时,对DNA具有较好的阻滞；当GHCn-m/DNA为1：1时,不对称阳离子脂质体对Hela细胞的转染效率的最高,比商品试剂DOTAP略高；对细胞毒性进行测试,发现不对称脂质体对Hela细胞的毒性较大。
[Abstract]:In recent years, the research of gene therapy has been one of the most important fields in the medical field. It has been used in clinic for tumor, immunodeficiency and other diseases, but its development is seriously affected by its high toxicity and relatively low efficiency. Compared with the low safety of viral vectors, the low immune response, low toxicity and easy preparation of non-viral vectors make it a hot research topic. In this paper, six kinds of cationic surfactants, GHCC12-14, GHCC14-16, GHBC12-14, GHBC12-16 and GHBC14-16, have been designed and synthesized. The long chain isocyanate was linked to the terminal hydroxyl of N (N) -dimethyl amino-1o 2-propanediol by 1H-1H correlation spectroscopy, and all the intermediates and target products were characterized by MS ~ (1H) NMR and 1H-1H gCOSY, and the structure was correct. The HLB values and critical permutation parameters P of the synthesized asymmetric cationic surfactants were investigated. It is found that the surface tension and cmc increase with the increase of carbon chain length. The 纬 cmc and cmc of surfactant with carbon chain length 12-14 are smaller and have better surface activity. Six kinds of symmetric cationic surfactants and six asymmetric cationic surfactants were combined with anionic surfactants SDBSU SDS and SAS to prepare vesicles, and their appearance, size and Zeta potential were analyzed. The results of conductivity and transmission electron microscopy (TEM) TEM show that vesicles can be formed in anion or anionic regions of anion / cationic surfactant composite system. Except for X _ 1 = 0.5, precipitation or sedimentation would occur, and the particle size distribution was almost 200 ~ 400 nm. The Zeta potential was less than -30 MV in anion rich region and more than 30 MV in Fuyang ion region, which indicated that the vesicle formed spontaneously in mixed solution had better stability. The mechanism of vesicle formation was explained by Masaniko geometric model and Israelachvili theory. Six kinds of asymmetric cationic surfactants with hydrophobic chain were studied. Tem showed that GHCn-m with asymmetric cationic liposomes could form liposomes except GHBC14-16. The study on the particle size and Zeta potential of asymmetric cationic liposomes combined with DNA showed that most of them were between 300nm and 400nm, which met the requirements of cell transfection. When the ratio of asymmetric cationic liposome GHCn-m to DNA was greater than 4:1, the inhibition of DNA was better. When GHCn-m/DNA was 1:1, the transfection efficiency of asymmetric cationic liposome to Hela cells was the highest, which was slightly higher than that of commercial reagent DOTAP. It was found that asymmetric liposomes were more toxic to Hela cells.
【学位授予单位】：大连理工大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R450;O647.2

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