以4-溴二苯甲酮、4,4'-二甲氧基二苯甲酮、对硝基苄溴(Br-BNO2)和叠氮基乳糖(LT)等为原料,经亲核取代、铃木偶联和click反应等构建了一种四苯乙烯(TPE)为骨架,并能高效产生活性氧(ROS)的光动力学治疗(PDT)试剂LT-TPE-DB-BNO2. 采用分光光度法和共聚焦荧光显微镜探究LT-TPE-DB-BNO2在血清和不同pH值溶液中的稳定性及细胞内外产生ROS的能力;利用MTT法和活死细胞双染法测定其对HeLa与BRL-3A细胞活力的影响. 结果表明: 在不同pH值溶液及血清中LT-TPE-DB-BNO2表现出优良的稳定性;在黑暗条件下,LT-TPE-DB-BNO2对HeLa和BRL-3A细胞的活力影响很小;在氙灯辐照下,LT-TPE-DB-BNO2高效产生ROS,使HeLa细胞的活力下降88.3%,表明LT-TPE-DB-BNO2是高效的PDT试剂.
A photodynamic therapy(PDT)reagent, LT-TPE-DB-BNO2 for reactive oxygen species(ROS)generation based on tetraphene(TPE), was synthesized from 4-bromobenzoylbenzene, 4,4’-dimethoxybenzophenone, p-nitrobenzyl bromide(Br-BNO2)and azide lactose(LT)by nucleophilic substitution, suzuki coupling and click reaction. The stability of LT-TPE-DB-BNO2 in serum and at different pH value, and the ability of ROS generation were investigated by spectrophotometry and confocal fluorescence microscopy. The effects of LT-TPE-DB-BNO2 on the viability of HeLa and BRL-3A cells were investigated by MTT method and live and dead cell double staining method. The results showed that LT-TPE-DB-BNO2 was stable in serum and at different pH value. In the dark condition,it had a little effect on the activity of HeLa and BRL-3A cells. Under irradiation by xenon lamp, ROS was produced by LT-TPE-DB-BNO2, and the activity of HeLa cells was reduced by 88.3%, indicating that LT-TPE-DB-BNO2 is a highly efficient PDT reagent.
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