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将-环糊精的2位羟基进行硒化得到的产物(2-SeCD)作为谷胱甘肽过氧化物酶(GPX)的一种人工模拟酶具有广阔的应用开发前景。为了对其进行进一步的研究及生产,需得到大量的产物。在原有的研究基础上,将中试放大实验的剂量扩大为小试实验的50倍,在7次工艺成熟的中试放大实验中获得了大量的2-SeCD。通过碳氢元素分析、红外光谱分析、核磁共振分析、质谱分析等表征实验验证了放大实验所得产物的结构正确,并测定其活力为5.2 U/mol。在2-SeCD对细胞的生物学效应实验中,发现在2-SeCD浓度较低的条件下可降低细胞死亡率而在浓度较高的条件下却抑制细胞生长。通过形态学观察与DNA ladder检测发现高浓度的2-SeCD可诱导Hela细胞凋亡,并经凋亡因子caspase-3的相对活力检测确定此凋亡是由线粒体caspase激活途径诱发的。
The product (2-SeCD) obtained by selenization of the 2-hydroxyl group of cyclodextrin has a broad application development prospect as an artificial mimic enzyme of glutathione peroxidase (GPX). In order to further its research and production, need to get a large number of products. Based on the previous studies, the dose of pilot scale-up experiment was expanded to 50 times of that of pilot-scale experiment. A large amount of 2-SeCD was obtained in seven pilot scale-up experiments. The structure of the product obtained by the amplification experiment was verified by the analysis of elemental analysis of hydrogen, infrared spectrum, nuclear magnetic resonance and mass spectrometry, and its activity was determined to be 5.2 U / mol. In experiments on the biological effects of 2-SeCD on cells, it was found that cell death was reduced at lower concentrations of 2-SeCD and inhibited cell growth at higher concentrations. Morphological observation and DNA ladder showed that high concentration of 2-SeCD induced apoptosis of Hela cells. The apoptosis of Hela cells was confirmed by the relative activity of caspase-3. The apoptosis was induced by mitochondrial caspase activation.