聚丙稀酸的氨基化修饰及其基因递送活性的研究

发布时间：2017-12-28 07:04

本文关键词：聚丙稀酸的氨基化修饰及其基因递送活性的研究　出处：《内蒙古大学》2017年硕士论文　论文类型：学位论文

【摘要】：基因治疗作为一种新型的医疗技术,在治疗肿瘤等疾病方面有很大的应用前景,因此受到了研究者们的关注。但是,由于基因载体的高毒性、低转染率等问题,使基因治疗在临床应用中受到了很大的限制。所以开发一种新型有效的基因载体一直是广大研究者的关注热点。阳离子高分子基因载体是人们研究最广泛的基因载体之一,针对其毒性高和转染效率低的缺点人们对其进行了一系列的修饰,改变N的类型是其中一种修饰方式。本论文主要研究了作为基因载体时不同氨基类型的化合物对生物活性的影响。本论文以聚丙烯酸为母体,通过酰胺反应连接不同N类型的支链对其进行氨基化修饰,两种不同的氨基分别选用脂肪链上的伯胺和丙氨酸上的α氨基,并通过改变这两种氨基的比例合成出四种化合物分别是PALA0、PALA1、PALA2、PALA3。对于合成出的化合物,我们通过氢谱、碳谱和质谱表征方法确定了最终结构。通过凝胶渗透色谱、激光粒度和电势电位测试得出:我们合成的化合物分子量在28000左右,粒径大小符合基因载体的最佳粒径,具有正的电势电位。酸碱滴定实验表明四种化合物都有很好的缓冲能力。本论文又通过琼脂糖凝胶电泳、细胞毒性实验、质粒转染、siRNA转染和FITC-DNA转染实验对化合物的生物活性进行了探究。结果表明,四种化合物都有很好的生物活性,与丙氨酸上的α氨基相比,脂肪链上的伯胺具有更好的细胞转染效果和细胞穿透能力,但是却有明显的细胞毒性。综上所述,我们成功对聚丙烯酸用不同类型的氨基进行了氨基化修饰,并对不同化合物作为基因载体时的生物活性进行了讨论,为以后研究者对基因载体的修饰提供了理论基础和研究依据。
[Abstract]:As a new kind of medical technology, gene therapy has a great prospect in the treatment of cancer and other diseases, so it has attracted the attention of researchers. However, because of the high toxicity and low transfection rate of gene carrier, gene therapy has been greatly restricted in clinical application. Therefore, the development of a new and effective gene carrier has been the focus of attention of the vast majority of researchers. Cationic polymer gene carrier is one of the most widely studied gene carriers. A series of modifications have been carried out for its disadvantages of high toxicity and low transfection efficiency. Changing the type of N is one of the modification methods. In this paper, the effects of different amino types on biological activity as gene carriers were studied. In this paper, polyacrylic acid as precursor by amidation reaction of different types of N branched connection of the amino modified, two different amino alpha amino fatty chain were selected on the primary amine and alanine on the two, and by changing the proportion of amino synthesized four kinds of compounds were PALA0, PALA1, PALA2, PALA3. For the synthesized compounds, we determine the final structure by the hydrogen spectrum, carbon spectrum and mass spectrometry. The gel permeation chromatography, laser particle size and potential potential test showed that the molecular weight of the synthesized compounds was about 28000, and the particle size accords with the best particle size of the gene vector and has positive potential potential. The acid base titration experiment shows that all the four compounds have good buffering ability. The biological activities of the compounds were also investigated by agarose gel electrophoresis, cytotoxicity, plasmid transfection, siRNA transfection and FITC-DNA transfection. The results showed that all four compounds had good bioactivity. Compared with the alpha amino group on alanine, the primary amine on the fat chain had better cell transfection efficiency and cell penetration ability, but it had obvious cytotoxicity. In conclusion, we succeeded in aminated modification of polyacrylic acid with different types of amino groups, and discussed the biological activity of different compounds as gene vectors, which provided theoretical basis and research basis for future researchers to modify gene carriers.
【学位授予单位】：内蒙古大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R450

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