4'修饰核苷类似物：强抑制剂有效对抗恩替耐药乙肝炎

StephenW

4′-modified nucleoside analogs: Potent inhibitors active against entecavir-resistant hepatitis B virus

    Yuki Takamatsu1,†, Yasuhito Tanaka2,†, Satoru Kohgo3,4, Shuko Murakami2, Kamalendra Singh5, Debananda Das1, David J. Venzon6, Masayuki Amano7, Nobuyo Higashi-Kuwata3,7, Manabu Aoki1,7,8, Nicole S. Delino1, Sanae Hayashi2, Satoru Takahashi9, Yoshikazu Sukenaga3, Kazuhiro Haraguchi4, Stefan G. Sarafianos5, Kenji Maeda1,3,* andHiroaki Mitsuya1,3,7

Article first published online: 25 AUG 2015

DOI: 10.1002/hep.27962

© 2015 by the American Association for the Study of Liver Diseases

Issue
Cover image for Vol. 62 Issue 3
Hepatology

    1    Experimental Retrovirology Section, HIV and AIDS Malignancy Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD
    2    Department of Virology & Liver Unit, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan
    3    Research Institute & Center for Clinical Sciences, National Center for Global Health and Medicine, Tokyo, Japan
    4    Department of Pharmaceutical Sciences, Nihon Pharmaceutical University, Saitama, Japan
    5    Molecular Microbiology & Immunology, Biochemistry, School of Medicine and Bond Life Sciences Center, University of Missouri, Columbia, MO
    6    Biostatistics and Data Management Section, National Cancer Institute, National Institutes of Health, Bethesda, MD
    7    Departments of Hematology, Rheumatology, and Clinical Immunology & Infectious Disease, Kumamoto University Graduate School of Medical Sciences, Kumamoto, Japan
    8    Department of Medical Technology, Kumamoto Health Science University, Kumamoto, Japan
    9    Department of Experimental Pathology & Tumor Biology, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan.

    †    Equally contributed in the present study.

*Address reprint requests to: Kenji Maeda, M.D., Ph.D., Center for Clinical Sciences, National Center for Global Health and Medicine, 1-21-1 Toyama, Shinjuku, Tokyo 162-8655, Japan. E-mail: [email protected]; fax: +81-3-3207-1038.

    Potential conflicts of interest: S.K. and H.M. are among the coinventors on a patent for 4′-ethynyl-2-fluoro-2′-deoxyadenosine; all the rights, title, and interest to the patent have been assigned to Yamasa Corporation, Chiba, Japan. Dr. Tanaka is on the speakers’ bureau for and received grants from Bristol-Myers Squibb. He received grants from Chugai and MSD.

    This work was supported, in part, by the Intramural Research Program of Center for Cancer Research, National Cancer Institute, National Institutes of Health (H.M.), in part by a Health and Labor Sciences Research Grant (Practical Research on Hepatitis [Research on the innovative development and the practical application of new drugs for hepatitis B [H.M., Ya.T., K.H.]), and in part by a Grant from International Research Center Aiming at the Control of AIDS, Kumamoto University (H.M.). S.G.S. was supported by R01AI112417 and R01AI076119.


Certain nucleoside/nucleotide reverse transcriptase (RT) inhibitors (NRTIs) are effective against human immunodeficiency virus type 1 (HIV-1) and hepatitis B virus (HBV). However, both viruses often acquire NRTI resistance, making it crucial to develop more-potent agents that offer profound viral suppression. Here, we report that 4′-C-cyano-2-amino-2′-deoxyadenosine (CAdA) is a novel, highly potent inhibitor of both HBV (half maximal inhibitory concentration [IC50] = 0.4 nM) and HIV-1 (IC50 = 0.4 nM). In contrast, the approved anti-HBV NRTI, entecavir (ETV), potently inhibits HBV (IC50 = 0.7 nM), but is much less active against HIV-1 (IC50 = 1,000 nM). Similarly, the highly potent HIV-1 inhibitor, 4′-ethynyl-2-fluoro-2′-deoxyadenosine (EFdA; IC50 = 0.3 nM) is less active against HBV (IC50 = 160 nM). Southern analysis using Huh-7 cells transfected with HBV-containing plasmids demonstrated that CAdA was potent against both wild-type (IC50 = 7.2 nM) and ETV-resistant HBV (IC50 = 69.6 nM for inline image), whereas ETV failed to reduce inline image DNA even at 1 μM. Once-daily peroral administration of CAdA reduced inline image viremia (P = 0.0005) in human-liver-chimeric/ inline image–infected mice, whereas ETV completely failed to reduce inline image viremia. None of the mice had significant drug-related body-weight or serum human-albumin concentration changes. Molecular modeling suggests that a shallower HBV-RT hydrophobic pocket at the polymerase active site can better accommodate the slightly shorter 4′-cyano of CAdA-triphosphate (TP), but not the longer 4′-ethynyl of EFdA-TP. In contrast, the deeper HIV-1-RT pocket can efficiently accommodate the 4′-substitutions of both NRTIs. The ETV-TP's cyclopentyl ring can bind more efficiently at the shallow HBV-RT binding pocket. Conclusion: These data provide insights on the structural and functional associations of HBV- and HIV-1-RTs and show that CAdA may offer new therapeutic options for HBV patients. (Hepatology 2015)

StephenW

4'修饰核苷类似物：强抑制剂有效对抗恩替卡韦耐药乙型肝炎病毒

    纪Takamatsu1，†，保仁Tanaka2，†，悟Kohgo3,4，修子Murakami2，Kamalendra Singh5，Debananda DAS1，戴维·Venzon6，雅之Amano7，Nobuyo东Kuwata3,7，押尾Aoki1,7,8，妮可S. Delino1 ，早苗Hayashi2，悟Takahashi9，嘉Sukenaga3，弘Haraguchi4，斯特凡G. Sarafianos5，贤治Maeda1,3，* andHiroaki Mitsuya1,3,7

文章首次在网上公布：2015年8月25日

DOI：10.1002 / hep.27962

©2015年肝病研究的美国协会

问题
封面图片卷。 62第3期
肝病

    1实验部分反转录病毒学，艾滋病毒和艾滋病的恶性科，美国国家癌症研究所，美国国立卫生研究院，马里兰州贝塞斯达
    病毒学和肝单位，医学科学名古屋市立大学研究生院，名古屋，日本的2部
    3研究所和中心临床科学，国家中心的全球健康和医学，日本东京
    药学院日本药科大学4系，埼玉县，日本
    5分子微生物学与免疫学，生物化学，医学和邦德生命科学中心，密苏里大学，哥伦比亚大学，密苏里州的学校
    6生物统计和信息管理部，美国国家癌症研究所，美国国立卫生研究院，马里兰州贝塞斯达
    7部门血液科，风湿和临床免疫学和传染病，医学科学熊本大学研究生院，熊本，日本
    医疗技术，熊本保健科学大学，熊本，日本的8部
    实验病理学和肿瘤生物学，医学科学名古屋市立大学研究生院，名古屋，日本的9部。

    †同样贡献在本研究中。

*地址转载请：前田贤治，医学博士，哲学博士，中心临床科学，国家中心的全球健康和医学，1-21-1富山县，东京都新宿区162-8655，日本。电子信箱：[email protected];传真：+ 81-3-3207-1038。

    潜在的利益冲突：S.K.和H.M.上一个专利的coinventors为4'-乙炔基-2-氟-2'-脱氧腺苷之间;所有权利，所有权和利益，以该专利已转让给山佐公司，日本千叶县。田中博士是扬声器的来自施贵宝局，并获得资助。他从中外制药公司和MSD的赠款。

    这项工作是支持的，部分由美国国家癌症研究所，健康（HM）国立研究所院内研究计划中心的癌症研究，在由卫生和劳动科学研究补助金（在实践研究肝炎[研究的一部分创新发展和新的药物对乙肝[HM，Ya.T.，KH]）的实际应用，并在由国际研究中心资助针对艾滋病的熊本大学的控制，（HM）的一部分。 S.G.S.由R01AI112417和R01AI076119支持。


某些核苷/核苷酸逆转录酶（RT）抑制剂（NRTI类药物）能有效对抗人类免疫缺陷病毒1型（HIV-1）和B型肝炎病毒（HBV）。然而，这两种病毒往往掌握NRTI阻力，使得它的关键，开发更有效的药物，提供着地抑制病毒。在这里，我们报告，4'-C-氰基-2-氨基-2'-脱氧腺苷（以下CAdA）是一种新型的，抑制剂两者的HBV（半最大抑制浓度[IC 50] = 0.4纳米）和HIV-1的高度有效的（ IC50 = 0.4纳米）。与此相反，经批准的抗HBV的NRTI，恩替卡韦（ETV），有效抑制乙肝病毒（IC 50 = 0.7纳米），但是小得多的活性抗HIV-1（IC 50 = 1000纳米）。类似地，高度有效的HIV-1的抑制剂，4'-乙炔基-2-氟-2'-脱氧腺苷（EFDA; IC 50 = 0.3 nm）的是对HBV（IC 50 = 160 nm）的活性较低。使用Huh-7细胞转染了含有HBV质粒Southern分析表明以下CAdA是有效的针对野生型（IC 50 = 7.2纳米）和ETV耐药HBV（IC 50 = 69.6纳米的内嵌图像），而ETV未能降低内嵌像DNA即使在1微米。以下CAdA每天一次口服给药在人类肝嵌合/内嵌图像感染小鼠减少内嵌图像血症（P = 0.0005），而ETV完全没有减少内嵌图像血症。无小鼠有显著毒品有关的体重或血清人白蛋白浓度的变化。分子模型表明，一个较浅的HBV-RT疏水口袋的聚合酶活性位点可更好地适应稍短4'-氰基CADA-三磷酸（TP）的，而不是长期4'-乙炔基EFDA-TP的。与此相反，在更深的HIV-1逆转录口袋可有效地容纳4'-取代两者的NRTIs。在ETV-TP的环戊基环可以在浅HBV-RT结合口袋结合更有效。结论：这些数据提供了对HBV-和HIV-1-RTS的结构和功能团体的见解，并表明可以下CAdA提供新的治疗选择乙肝患者。 （肝病2015年）