原卟啉钠的制备及其体外抗乙型肝炎病毒作用的实验性研究

目的:提取原卟啉钠并体外观察其抗乙肝病毒的作用及其细胞毒性作用。方法:从抗凝猪血中提取血红素,经氯化血红素、原卟啉二甲酯等中间制备过程,最后制成原卟啉二钠。将NAPP水溶液160μg/m l,80μg/m l,40μg/m l,20μg/m l,10μg/m l分别加入H epG 2.2.15细胞株培养悬液中,8d后检测上清液中HBeA g的含量,并计算HBeA g抑制率、细胞存活率、HBeA g抑制率为50%时的药物浓度、实验组存活细胞为对照组的50%时的药物浓度和治疗指数。结果:所制得的NAPP为紫褐色晶体,可溶于水和甲醇,不溶于氯仿、乙醚、丙酮,难溶于稀酸,纯度为91.4%。当加人的NAPP浓度为160μg/m l和80μg/m l时,HBeA g抑制率分别为73.45%和37.25%,细胞存活率分别为96.30%和97.35%。NAPP对分泌HBeA g的治疗指数为7.23。结论:这是一种较理想的制备NAPP方法,且NAPP可有效抑制HBV-DNA的表达和复制,对靶细胞(肝细胞)无细胞毒作用。

Preparation of protoporphyroin disodium and in-vitro inhibitory effect of napp on hepatitis b virus

Objective :To study simple preparation of protoporphyrin disodium and to observe the in-vitro inhibitory effect of NAPP on Hepatitis B virus and its cytotoxicity to 2.2.15 cell strain.Methods : Esterification of hemin was undertaken to produce crude protoporphyrin dimethyl ester which was purified in toluene and then saponified by sodium hydroxide.The different concentration(160μg/ml,80μg/ml,40μg/m1,20μg/ml,10μg/ml) of NAPP water solution was respectively added into cell culture fluid of 2.2.15 cell strain and the inhibitory effect of NAPP on HBeAg was evaluated.Results :The protoporphyrin disodium obtained,the purity in 91.4%,was a dark brown red solids It was soluble in water,acidic solution and insoluble in ethyl ether.When drug concentration of NAPP is 160μg/ml,80μg/ml,inhibitive rates of HBeAg were 73.45% and 37.25 %,cellar survival rates were 96.30% and 97.35% respectively.The therapeutic index(the ratio of 50% toxic concentration and 50% inhibitory concentration) of NAPP was 7.23 for HBeAg.Conclusion :This is an ideal approach to prepare protoporphyrin disodium and NAPP inhibit duplication of Hepatitis B Virus in Vitro.