Edwards Tiffany C, Lomonosova Elena, Patel Jenny A, Li Qilan, Villa Juan A, Gupta Ankit K, Morrison Lynda A, Bailly Fabrice, Cotelle Philippe, Giannakopoulou Erofili, Zoidis Grigoris, Tavis John E
Department of Molecular Microbiology and Immunology and Saint Louis University Liver Center, Saint Louis University School of Medicine, St. Louis, MO, USA; Saint Louis University Liver Center, Saint Louis University School of Medicine, St. Louis, MO, USA.
Department of Molecular Microbiology and Immunology and Saint Louis University Liver Center, Saint Louis University School of Medicine, St. Louis, MO, USA.
Antiviral Res. 2017 Jul;143:205-217. doi: 10.1016/j.antiviral.2017.04.012. Epub 2017 Apr 25.
We previously reported low sensitivity of the hepatitis B virus (HBV) ribonuclease H (RNaseH) enzyme to inhibition by N-hydroxyisoquinolinedione (HID) compounds. Subsequently, our biochemical RNaseH assay was found to have a high false negative rate for predicting HBV replication inhibition, leading to underestimation of the number of HIDs that inhibit HBV replication. Here, 39 HID compounds and structurally related polyoxygenated heterocycles (POH), N-hydroxypyridinediones (HPD), and flutimides were screened for inhibition of HBV replication in vitro. Inhibiting the HBV RNaseH preferentially blocks synthesis of the positive-polarity DNA strand and causes accumulation of RNA:DNA heteroduplexes. Eleven HIDs and one HPD preferentially inhibited HBV positive-polarity DNA strand accumulation. ECs ranged from 0.69 μM to 19 μM with therapeutic indices from 2.4 to 71. Neither the HIDs nor the HPD had an effect on the ability of the polymerase to elongate DNA strands in capsids. HBV RNaseH inhibition by the HIDs was confirmed with an improved RNaseH assay and by detecting accumulation RNA:DNA heteroduplexes in HBV capsids from cells treated with a representative HID. Therefore, the HID scaffold is more promising for anti-HBV drug discovery than we originally reported, and the HPD scaffold may hold potential for antiviral development. The preliminary structure-activity relationship will guide optimization of the HID/HPDs as HBV inhibitors.
我们之前报道过，乙型肝炎病毒（HBV）核糖核酸酶H（RNaseH）对N-羟基异喹啉二酮（HID）化合物的抑制作用敏感性较低。随后，我们发现生化RNaseH检测在预测HBV复制抑制方面存在较高的假阴性率，导致对抑制HBV复制的HID数量估计不足。在此，对39种HID化合物以及结构相关的多氧化杂环（POH）、N-羟基吡啶二酮（HPD）和氟替米特进行了体外抑制HBV复制的筛选。抑制HBV RNaseH优先阻断正链DNA的合成，并导致RNA:DNA杂合双链的积累。11种HID和1种HPD优先抑制HBV正链DNA的积累。半数有效浓度范围为0.69μM至19μM，治疗指数为2.4至71。HID和HPD对衣壳中DNA链延伸的聚合酶能力均无影响。通过改进的RNaseH检测以及检测用代表性HID处理的细胞中HBV衣壳内RNA:DNA杂合双链的积累，证实了HID对HBV RNaseH的抑制作用。因此，HID骨架在抗HBV药物研发方面比我们最初报道的更具前景，HPD骨架可能具有抗病毒开发潜力。初步的构效关系将指导HID/HPD作为HBV抑制剂的优化。
