乙肝病毒复制的通过靶向核苷酸还原酶M2蛋白抑制

StephenW

Biochem Pharmacol. 2016 Jan 13. pii: S0006-2952(16)00020-4. doi: 10.1016/j.bcp.2016.01.003. [Epub ahead of print]
Inhibition of hepatitis B virus replication by targeting ribonucleotide reductase M2 protein.Liu X1, Xu Z2, Hou C2, Wang M2, Chen X3, Lin Q4, Song R4, Lou M4, Zhu L4, Qiu Y5, Chen Z5, Yang C6, Zhu W7, Shao J8.
Author information

• 1Department of Pathology and Pathophysiology, Key Laboratory of Disease Proteomics of Zhejiang Province, Research Center for Air Pollution and Health, Zhejiang University School of Medicine, Hangzhou 310058, China; Central Laboratory, The Second Clinical Medical College, Zhejiang Chinese Medical University, Hangzhou, China.
• 2Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China.
• 3Department of Pathology and Pathophysiology, Key Laboratory of Disease Proteomics of Zhejiang Province, Research Center for Air Pollution and Health, Zhejiang University School of Medicine, Hangzhou 310058, China; Department of Pathology and Pathophysiology, Zhengzhou University School of Medicine, Zhengzhou 450001, China.
• 4Department of Pathology and Pathophysiology, Key Laboratory of Disease Proteomics of Zhejiang Province, Research Center for Air Pollution and Health, Zhejiang University School of Medicine, Hangzhou 310058, China.
• 5State Key Laboratory of Infectious Disease Diagnosis and Treatment, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, China.
• 6Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China. Electronic address: [email protected].
• 7Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China. Electronic address: [email protected].
• 8Department of Pathology and Pathophysiology, Key Laboratory of Disease Proteomics of Zhejiang Province, Research Center for Air Pollution and Health, Zhejiang University School of Medicine, Hangzhou 310058, China. Electronic address: [email protected].
  

AbstractChronic 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-cyclopropyl-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.
Copyright © 2016. Published by Elsevier Inc.


KEYWORDS: Drug synergism and Drug resistance; Hepatitis B virus; Human ribonucleotide Reductase small subunit M2 (RRM2); Small molecule compounds; Viral genomic DNA and cccDNA synthesis

StephenW

生化药理学。 2016年13月PII：S0006-2952（16）00020-4。 DOI：10.1016 / j.bcp.2016.01.003。 [打印EPUB提前]
乙肝病毒复制的通过靶向核苷酸还原酶M2蛋白抑制。
刘X1，徐Z2，侯C2，王M2，陈X3，林Q4，宋R4，娄M4，朱L4，邱Y5，陈Z5，杨C6，朱W7，邵J8。
作者信息

    教研室病理学与病理生理学，浙江省，研究中心空气污染与健康，浙江大学医学院，杭州310058，中国疾病蛋白质组学重点实验室;中心实验室，第二临床医学院，浙江中国医药大学，中国杭州。
    本草，中国院士，上海，201203，中国2上海市的研究所。
    3Department病理学与病理生理学，浙江省，研究中心空气污染与健康，浙江大学医学院，杭州310058，中国疾病蛋白质组学重点实验室;病理学与病理生理学，郑州大学医学院，郑州450001，中国系。
    4Department病理学与病理生理学，浙江省，研究中心空气污染与健康，浙江大学医学院，杭州310058，中国疾病蛋白质组学重点实验室。
    感染性疾病的诊断和治疗，第一附属医院，医学院，浙江大学，杭州310003，中国的5State重点实验室。
    本草，中国院士，上海，201203，中国的6Shanghai研究所。电子地址：[email protected]。
    本草，中国院士，上海，201203，中国的7Shanghai研究所。电子地址：[email protected]。
    8Department病理学与病理生理学，浙江省，研究中心空气污染与健康，浙江大学医学院，杭州310058，中国疾病蛋白质组学重点实验室。电子地址：[email protected]。

抽象

慢性乙型肝炎病毒（HBV）感染是世界范围肝细胞癌的关键因素。核糖核苷酸还原酶（RR）调节脱氧核苷三磷酸的生物合成，并作为抗癌治疗的靶。这里，我们表明，RR是对HBV复制和病毒共价闭合环状DNA（cccDNA的）的合成在宿主肝细胞是必不可少的。通过执行计算机辅助虚拟筛选针对的RR小亚基M2（RRM2），osalmid的晶体结构，被鉴定为潜在RRM2靶向化合物。 Osalmid被证明是10倍的活性在抑制RR活性比羟基脲，和显著抑制HBV DNA和cccDNA的合成在HepG2.2.15细胞。与此相反，羟基脲和RR大亚基（RRM1​​） - 抑制药物吉西他滨表现出抗乙肝病毒复制的小选择性的活动。此外，osalmid也被证明具有有效的活性针对3TC耐药HBV应变，这表明在治疗药物耐药HBV感染效用。有趣的是，osalmid表现出与拉米夫定（3TC）在体外和体内没有显著毒性的协同作用，并显示抑制体内RR活性，从而验证其在体内的功能。此外，4-环丙基-2-氟-N-（4-羟基苯基）苯甲酰胺（YZ51），一种新型osalmid的衍生物，表现出比osalmid与更有效的RR抑制活性功效更高。这些结果表明，RRM2可能被靶向HBV的抑制性，且RRM2靶向化合物osalmid及其衍生物YZ51可以是一类新的抗HBV候选人为乙型肝炎和HBV相关的肝癌治疗的潜在用途。

版权所有©2016年出版由Elsevier公司
关键词：

药物协同作用和耐药性;乙型肝炎病毒;人核糖核苷酸还原酶小亚基M2（RRM2）;小分子化合物;病毒基因组DNA和合成的cccDNA