一种新型MCC-478衍生物抗乙型肝炎病毒活性及体外毒性实验

doi:国家自然科学基金资助课题（30400566）；吉

一种新型MCC-478衍生物抗乙型肝炎病毒活性及体外毒性实验

吴荻¹，牛俊奇¹，吴新宇²，丁艳华¹，仲伯华³，冯相伟¹

1.吉林大学第一医院感染症科，吉林长春 130021；2.吉林大学公共卫生学院卫生化学教研室，吉林长春 130021；3.军事医学科学院毒物药物研究所，北京 100850

收稿日期:2006-10-20 修回日期:1900-01-01 出版日期:2007-05-28 发布日期:2007-05-28
通讯作者: 牛俊奇

Anti-hepatitis B virus activity and toxicity of a novel nucleoside analogues in vitro

WU Di¹, NIU Jun-qi¹, WU Xin-yu², DING Yan-hua¹, ZHONG Bo-hua³, FENG Xiang-wei¹

1.Department of Infectious Disease, First Hospital, Jilin University, Changchun 130021, China; 2. Department of Sanitary Chemistry, School of Public Health, Jilin University, Changchun 130021, China; 3.Institute of Poison and Drugs, Academy of Military Medical Sciences, Beijing 100850, China

Received:2006-10-20 Revised:1900-01-01 Online:2007-05-28 Published:2007-05-28
Contact: NIU Jun-qi

摘要/Abstract

摘要： 目的：研究一种具有新结构类型的MCC-478衍生物030705的抗乙肝病毒活性和体外毒性。方法：实验组以不同浓度受试化合物030705（0.01、0.03、0.10、0.30和1.0 μmol•L^-1）、对照组以不同浓度阿德福韦酯（0.10、0.30、1.0、3.0和10.0 μ mol•L^-1），分别作用于HepG2.2.15细胞，采用Southern blotting杂交法测定其对HBV DNA的抑制率，计算其50%抑制浓度（IC₅₀ ）和90%抑制浓（IC₉₀）值，采用ELISA法测定不同药物浓度受试化合物030705对HBeAg分泌的抑制率。采用MTT法测定不同浓度受试化合物030705（10、30、100、300和1 000　μmol•L^-1）对HepG2细胞毒性，计算其50%致死浓度（CC₅₀）值。采用Dot blotting法分别测定不同浓度受试化合物030705（0.10、1.0和10.0　μmol•L^-1）对HepG2细胞线粒体含量的抑制率；同时设相应浓度双脱氧胞苷（ddC）阳性药物对照组和仅加入培养基的阴性对照组。结果：受试化合物030705抑制HepG2.2.15细胞HBV DNA作用与阿德福韦酯对照组比较差异无显著性（P>0.05），显示其具有与阿德福韦酯相近的抗乙肝病毒活性。不同浓度受试化合物030705（0.01、0.03、0.10、0.30和1.0 μmol•L^-1）对HBeAg分泌的抑制率分别为5.94%、6.08%、6.32%、10.31%和12.49%，与阴性对照组比较差异均无显著性（P>0.05）。受试化合物030705对HepG2细胞毒性的CC₅₀值为2 014 μmol•L^-1(>1 000 μmol•L^-1)，属于低细胞毒性药物。阳性对照药物ddC在不同浓度下（0.10、1.0和10.0 μmol•L^-1），对HepG2细胞线粒体含量的抑制率分别为38.43%、46.51 %、56.51%，与阴性对照组比较差异均有显著性（P＜0.05）。相同浓度下受试化合物030705抑制率分别为7.00%、5.81%、5.78%，与阴性对照组比较差异均无显著性（P>0.05）。结论：受试化合物030705对HepG2.2.15细胞HBV DNA的抑制作用与阿德福韦酯的作用相近，对HepG2细胞毒性的CC₅₀值2 014 μmol•L^-1，无明显线粒体毒性，是一种低毒、高效的新型核苷类抗乙肝病毒药物。

关键词: 肝炎, 乙型, 慢性, 药物疗法

Abstract: To study the anti-HBV activity and toxicity of a novel MCC-478 devivative 030705 with specific structure in vitro. Methods Anti-HBV activities in HepG2.2.15 cells were analyzed by Southern blotting in experimental groups with different doses of 030705 （0.01, 0.03, 0.10, 0.30 and 1.0 μmol•L^-1）and Adefovir Dipioxil positive control groups(0.10, 0.30, 1.0, 3.0 and 10.0 μmol•L^-1). Concentration of 50% inhibitation (IC₅₀)and concentration of 90% inhibitation (IC₉₀) were obtained. The inhibitory effect of HBeAg was analyzed in HepG2.2.15 cells by ELISA method. The cytotoxicities were tested by MTT method in the experimental groups（10, 30, 100, 300 and 1 000 μmol•L^-1） in HepG2 cells. Concentration of 50% extinction (CC₅₀) was obtained. The inhibitory effects of mitochondrial DNA contents in experimental groups（0.10, 1.0 and 10 μmol•L^-1）were tested by Dot blotting in HepG2 cells. While didenoxycytidine (ddC) groups were as positive control groups and untreated groups were as negative groups. Results Compared with Adefovir Dipioxil positive control groups, the inhibitory effects of the test compound 030705 on HBV DNA had no obvious difference（P>0.05）. Anti-HBV activity of the test compound 030705 was similar to that of Adefovir Dipioxil in HepG2. 2.15 cells. The inhibitory rates of HBeAg in experimental groups （0.01, 0.03, 0.10, 0.30 and 1.0 μmol•L^-1 030705）were 5.94%, 6.08%, 6.32%, 10.31% and 12.49%, respectively in HepG2.2.15 cells; compared with negative control group, there was no obvious difference（P>0.05）. CC₅₀ value of the test compound was 2 014 μmol•L^-1 (>1 000 μmol•L^-1) in HepG2 cells. The inhibitory rates of mitochondrial DNA content from positive drug ddC （0.10, 1.0 and 10 μmol•L^-1）were 38.43%,46.51%,56.51%, respectively in HepG2 cells; compared with negative control group, there was obvious difference（P＜0.05）. The inhibitory rates of mitochondrial DNA content from the test compound 030705 with the same concentrations were 7.00%,5.81%,5.78%, respectively in HepG2 cells; compared with negative control group, there was no obvious difference（P＞0.05）. Conclusion Anti-HBV activity of the test compound 030705 was similar to that of Adefovir Dipioxil in HepG2.2.15 cells. CC₅₀ value of the test compound is 2014 μmol•L^-1 (>1 000 μmol•L^-1) in HepG2 cells. No apparent reductions of mitochondrial DNA content are observed in HepG2 cells. The test compound 030705 with specific structure may be a new promising anti-HBV agent.

Key words: hepatitis B, chronic, drug therapy

中图分类号:

R512.6

吴荻，牛俊奇，吴新宇，丁艳华，仲伯华，冯相伟. 一种新型MCC-478衍生物抗乙型肝炎病毒活性及体外毒性实验[J]. J4, 2007, 33(3): 422-426.

WU Di, NIU Jun-qi, WU Xin-yu, DING Yan-hua, ZHONG Bo-hua, FENG Xiang-wei. Anti-hepatitis B virus activity and toxicity of a novel nucleoside analogues in vitro[J]. J4, 2007, 33(3): 422-426.

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