Inhibition of hepatitis B virus replication by targeting ribonucleotide reductase M2 protein
Inhibition of hepatitis B virus replication by targeting ribonucleotide reductase M2 protein
复制标题
通过靶向核糖核苷酸还原酶 M2 蛋白抑制乙型肝炎病毒复制
DOI：
10.1016/j.bcp.2016.01.003
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发表时间：
2016-03-01
影响因子：
5.8
通讯作者：
Shao, Jimin
中科院分区：
文献类型：
--
作者：
Liu, Xia;Xu, Zhijian;Shao, Jimin
慢性B型肝炎病毒（HBV）感染是世界范围内肝细胞癌的关键因素。核糖核苷酸还原酶（ribonucleotide reductase，RR）调节脱氧核苷三磷酸的生物合成，是抗癌治疗的靶点。在这里，我们证明RR是必不可少的HBV复制和病毒共价闭合环状DNA（cccDNA）在宿主肝细胞的合成。通过对RR小亚基M2（RRM 2）的晶体结构进行计算机辅助虚拟筛选，确定osalmid为潜在的RRM 2靶向化合物。在HepG2.2.15细胞中，Osalmid在抑制RR活性方面的活性比羟基脲高10倍，并且显著抑制HBV DNA和cccDNA的合成。相反，羟基脲和RR大亚基（RRM 1）抑制药物吉西他滨对HBV复制的选择性活性很小。此外，osalmid还显示出对3 TC耐药HBV株具有有效活性，表明其在治疗耐药HBV感染中的实用性。有趣的是，奥沙米特在体外和体内与拉米夫定（3 TC）显示出协同作用，而没有明显的毒性，并显示出抑制RR活性，从而验证了其在体内的功能。此外，4-环丙基-2-氟-N-（4-羟基苯基）苯甲酰胺（YZ 51），一种新的衍生物，显示出更强的RR抑制活性比osalmid的效果。这些结果表明，RRM 2可能是HBV抑制的靶点，并且RRM 2靶向化合物osalmid及其衍生物YZ 51可能是一类新的抗HBV候选物，具有潜在的用于治疗B肝炎和HBV相关的HCC的用途。(C)2016 Elsevier Inc. All rights reserved.
Chronic hepatitis B virus (HBV) infection is a key factor for hepatocellular carcinoma worldwide. Ribonucleotide reductase (RR) regulates the deoxyribonucleoside triphosphates biosynthesis and serves as a target for anti-cancer therapy. Here, we demonstrate that RR is essential for HBV replication and the viral covalently-closed-circular DNA (cccDNA) synthesis in host liver cells. By performing computer-assisted virtual screening against the crystal structure of RR small subunit M2 (RRM2), osalmid, was identified as a potential RRM2-targeting compound. Osalmid was shown to be 10-fold more active in inhibiting RR activity than hydroxyurea, and significantly inhibited HBV DNA and cccDNA synthesis in HepG2.2.15 cells. In contrast, hydroxyurea and the RR large subunit (RRM1)-inhibitory drug gemcitabine showed little selective activity against HBV replication. In addition, osalmid also was shown to possess potent activity against a 3TC-resistant HBV strain, suggesting utility in treating drug-resistant HBV infections. Interestingly, osalmid showed synergistic effects with lamivudine (3TC) in vitro and, in vivo without significant toxicity, and was shown to inhibit RR activity in vivo, thus verifying its in vivo function. Furthermore, 4-cyclopropy1-2-fluoro-N-(4-hydroxyphenyl) benzamide (YZ51), a novel derivative of osalmid, showed higher efficacy than osalmid with more potent RR inhibitory activity. These results suggest that RRM2 might be targeted for HBV inhibition, and the RRM2-targeting compound osalmid and its derivative YZ51 could be a novel class of anti-HBV candidates with potential use for hepatitis B and HBV-related HCC treatment. (C) 2016 Elsevier Inc. All rights reserved.