Isothiafludine, a novel non-nucleoside compound, inhibits hepatitis B virus replication through blocking pregenomic RNA encapsidation  

作　　者：Li YANG Li-ping SHI Hai-jun CHEN Xian-kun TONG Gui-feng WANG Yang-ming ZHANG Wen-long WANG Chun-lan FENG Pei-lan HE Feng-hua ZHU You-hua HAO Bao-ju WANG Dong-liang YANG Wei TANG Fa-jun NAN Jian-ping ZUO 

机构地区：[1]Laboratory of Immunopharmacology [2]Chinese National Center for Drug Screening, State Key Laboratory of Drug Research, Shang- hai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China [3]Tongji Hospital of TonEji Medical College, Wuhan 430030, China

出　　处：《Acta Pharmacologica Sinica》2014年第3期410-418,共9页中国药理学报（英文版）

摘　　要：Aim： To investigate the action of isothiafludine （NZ-4）, a derivative of bis-heterocycle tandem pairs from the natural product leucamide A, on the replication cycle of hepatitis B virus （HBV） in vitro and in vivo. Methods： HBV replication cycle was monitored in HepG2.2.15 cells using qPCR, qRT-PCR, and Southern and Northern blotting. HBV protein expression and capsid assembly were detected using Western blotting and native agarose gel electrophoresis analysis. The interaction of pregenomic RNA （pgRNA） and the core protein was investigated by RNA immunoprecipitation. To evaluate the anti-HBV effect of NZ-4 in vivo, DHBV-infected ducks were orally administered NZ-4 （25, 50 or 100 mg.k^l{1-1） for 15 d. Results： NZ-4 suppressed intracellular HBV replication in HepG2.2.15 cells with an IC^o value of 1.33 pmol/L, whereas the compound inhibited the cell viability with an IC5o value of 50.4 pmol/L. Furthermore, NZ-4 was active against the replication of various drug- resistant HBV mutants, including 3TC/ETV-dual-resistant and ADV-resistant HBV mutants. NZ-4 （5, 10, and 20 pmol/L） concentration- dependently reduced the encapsidated HBV pgRNA, resulting in the assembly of replication-deficient capsids in HepG2.2.15 cells. Oral administration of NZ-4 dose-dependently inhibited DHBV DNA replication in the DHBV-infected ducks. Conclusion： NZ-4 inhibits HBV replication by interfering with the interaction between pgRNA and HBcAg in the capsid assembly process thus increasing the replication-deficient HBV capsids. Such mechanism of action might provide a new therapeutic strategy to combat H BV infection.Aim： To investigate the action of isothiafludine （NZ-4）, a derivative of bis-heterocycle tandem pairs from the natural product leucamide A, on the replication cycle of hepatitis B virus （HBV） in vitro and in vivo. Methods： HBV replication cycle was monitored in HepG2.2.15 cells using qPCR, qRT-PCR, and Southern and Northern blotting. HBV protein expression and capsid assembly were detected using Western blotting and native agarose gel electrophoresis analysis. The interaction of pregenomic RNA （pgRNA） and the core protein was investigated by RNA immunoprecipitation. To evaluate the anti-HBV effect of NZ-4 in vivo, DHBV-infected ducks were orally administered NZ-4 （25, 50 or 100 mg.k^l{1-1） for 15 d. Results： NZ-4 suppressed intracellular HBV replication in HepG2.2.15 cells with an IC^o value of 1.33 pmol/L, whereas the compound inhibited the cell viability with an IC5o value of 50.4 pmol/L. Furthermore, NZ-4 was active against the replication of various drug- resistant HBV mutants, including 3TC/ETV-dual-resistant and ADV-resistant HBV mutants. NZ-4 （5, 10, and 20 pmol/L） concentration- dependently reduced the encapsidated HBV pgRNA, resulting in the assembly of replication-deficient capsids in HepG2.2.15 cells. Oral administration of NZ-4 dose-dependently inhibited DHBV DNA replication in the DHBV-infected ducks. Conclusion： NZ-4 inhibits HBV replication by interfering with the interaction between pgRNA and HBcAg in the capsid assembly process thus increasing the replication-deficient HBV capsids. Such mechanism of action might provide a new therapeutic strategy to combat H BV infection.

关 键 词：hepatitis B virus replication capsid assembly pregenomic RNA non-nucleoside compound isothiafludine lamivudine ADEFOVIR ENTECAVIR 

分 类 号：S154.34[农业科学—土壤学] TQ464.6[农业科学—农业基础科学]