Chronic infection with hepatitis B virus (HBV) sometime induces lethal cirrhosis and hepatocellular carcinoma. Although nucleot(s)ide analogs are used as main treatment for HBV infection, the emergence of the drug‐resistant viruses has become a problem. To discover novel antivirals with low side effects and low risk of emergence of resistant viruses, screening for anti‐HBV compounds was performed with compound libraries of inhibitors targeting G‐protein‐coupled receptors (GPCRs). HepG2‐hNTCP C4 cells infected with HBV were treated with various GPCR inhibitors and harvested at 14 day postinfection for quantification of core protein in the first screening or relaxed circular DNA in the second screening. Finally, we identified a cannabinoid receptor 1 inhibitor, rimonabant, as a candidate showing anti‐HBV effect. In HepG2‐hNTCP C4 cells, treatment with rimonabant suppressed HBV propagation at the viral RNA transcription step but had no effect on entry or covalently closed circular DNA level. The values of half maximal inhibitory concentration, half maximal effective concentration, and selectivity index of rimonabant in primary human hepatocyte (PHH) are 2.77 μm, 40.4 μm, and 14.6, respectively. Transcriptome analysis of rimonabant‐treated primary hepatocytes by RNA sequencing revealed that the transcriptional activity of hepatocyte nuclear factor 4α (HNF4α), which is known to stimulate viral RNA synthesis, was depressed. By treatment of PHH with rimonabant, the expression level of HNF4α protein and the production of the messenger RNAs (mRNAs) of downstream factors promoted by HNF4α were reduced while the amount of HNF4α mRNA was not altered. These results suggest that treatment with rimonabant suppresses HBV propagation through the inhibition of HNF4α activity.

中文翻译：

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利莫那班通过抑制肝细胞核因子4α抑制乙型肝炎病毒的RNA转录。

乙型肝炎病毒（HBV）的慢性感染有时会导致致命性肝硬化和肝细胞癌。尽管核苷酸类似物被用作HBV感染的主要治疗方法，但耐药病毒的出现已成为一个问题。为了发现具有低副作用和低耐药病毒出现风险的新型抗病毒药物，使用针对G蛋白偶联受体（GPCR）的抑制剂化合物库对抗HBV化合物进行了筛选。用各种GPCR抑制剂处理感染HBV的HepG2-hNTCP C4细胞，并在感染后第14天收获，以在第一次筛选中定量核心蛋白或在第二次筛选中定量松弛的环状DNA。最后，我们确定了大麻素受体1抑制剂利莫那班作为显示抗HBV作用的候选药物。在HepG2-hNTCP C4细胞中，利莫那班治疗在病毒RNA转录步骤抑制了HBV传播，但对进入或共价闭合的环状DNA水平没有影响。原发性人类肝细胞（PHH）中利莫那班的最大抑制浓度的一半，有效最大抑制浓度的一半和选择性指数分别为2.77μm，40.4μm和14.6。通过RNA测序对利莫那班处理过的原代肝细胞进行的转录组分析显示，已知能刺激病毒RNA合成的肝细胞核因子4α（HNF4α）的转录活性受到抑制。通过利莫那班处理PHH，减少了HNF4α蛋白的表达水平和由HNF4α促进的下游因子的信使RNA（mRNA）的产生，而HNF4αmRNA的量没有改变。