【作者】 格日乐；

【导师】 胡日查；

【作者基本信息】 内蒙古大学 ， 应用化学， 2017， 硕士

【摘要】 RNA干扰(RNAi)是双连RNA诱导的转录后水平上的基因调控,把特异性靶mRNA降解,从而下调疾病有关的基因。它已经表现出了治疗基因的异常表达导致的各种疾病的巨大潜力。在RNAi的临床应用中,研究出高生物相容性、高效率递送载体极为关键。本实验室前期研究工作中,已经制备出的核酸载体有DoGo2、DoGo3、DoGo4α2δ2、DoGo4α3δ1脂质复合物等。它们都通过调控DoGo1的亲水头部而合成出来的。本论文以天然氨基酸原料构建了三种脂质化类肽,经过调节氨基酸组成、多肽氨基酸数、胺类合成出DoGo6、DoGo311、DoGo121。我们通过1H-NMR和质谱物性确定化合物的结构与分子量等,通过透射电镜、电动电势及激光粒度等方法对脂质复合物进行表征。我们对DoGo6、DoGo311、DoGo121脂质复合物的生物活性进行研究。实验结果表明,它们的生物相容性均比商用的转染试剂Lipofectamine 2000更好。DoGo6、DoGo311脂质复合物在较低的氨基(N)/磷酸基团(P)比例下可以与dsDNA结合。在体外转染实验中,转染质粒、siRNA(siGAPDH、siPLK1)。实验结果表明,随α氨基数的增加,能够增加DNA/siRNA转染效率,同时,DoGo6脂质复合物对293T、HeLa、HepG2等细胞株具有潜在的靶向性。更多还原

【Abstract】 RNA interference(RNAi)is a double-stranded RNA induced gene regulation at the post-transcriptional level,degrading the specific target mRNA,thereby down-regulating the disease-related genes.It has shown great potential for the curing of various diseases caused by the abnormal gene expression.In the clinical application of RNAi,the development of highly biocompatible,high-efficiency delivery vector is critical.Previously our laboratory has synthesized the potential nucleic acid carriers,namely DoGo2,DoGo3,DoGo4αa2δ2,DoGo4αa3δ1 lipoplex and so on.They are synthesized by modifying the hydrophilic head of DoGo1.In this paper,three kinds of lipid functionalized Peptidomimetics were synthesized from natural amine acid,and the impact of amino acid composition,amino acid number and the type of amine on their delivery efficiency were studied.The structures and molecular weights of the compounds(DoGo6,DoGo311,DoGol21)were determined by 1H-NMR and MS.The lipoplex were characterized by transmission electron microscopy,electromotive force and laser particle size.We studied the biological activity of DoGo6,DoGo311,DoGo121 lipoplex.The experimental results show that their biocompatibility is better than the commercial transfection reagent Lipofectamine 2000.DoGo6,DoGo311 lipoplex can bind to dsDNA at a lower phosphoric acid group(P)/amino(N)ratio.In vitro transfection experiments incldued plasmids transfection as well as siRNA(siGAPDH,siPLKl)transfection.The results showed that the transfection efficiency of DNA/siRNA was increased with the increasing number of amino acids in the head group,and the DoGo6 lipid complex had highest potential for targeting various cells including 293T,HeLa and HepG2 cells.更多还原