Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Second Affiliated Hospital, Chongqing Medical University, 1 Yi Xue Yuan Road, Yuzhong District, Chongqing 400016, People's Republic of China; Department of Molecular Microbiology and Immunology, Saint Louis University School of Medicine, 1100 S. Grand Blvd., Saint Louis, MO 63122, USA.
Antiviral Res. 2013 Sep;99(3):221-9. doi: 10.1016/j.antiviral.2013.06.007. Epub 2013 Jun 21.
Hepatitis B virus (HBV) is a hepatotropic DNA virus that replicates by reverse transcription. It chronically infects >350 million people and kills about 1 million patients annually. Therapy primarily employs nucleos(t)ide analogs that suppress viral DNA synthesis by the viral reverse transcriptase very well but that rarely cure the infection, so additional therapies are needed. Reverse transcription requires the viral ribonuclease H (RNAseH) to destroy the viral RNA after it has been copied into DNA. We recently produced active recombinant HBV RNAseH and demonstrated that Human Immunodeficiency Virus (HIV) RNAseH antagonists could inhibit the HBV enzyme at a high frequency. Here, we extended these results to β-thujaplicinol, a hydroxylated tropolone which inhibits the HIV RNAseH. β-Thujaplicinol inhibited RNAseHs from HBV genotype D and H in biochemical assays with IC₅₀ values of 5.9±0.7 and 2.3±1.7 μM, respectively. It blocked replication of HBV genotypes A and D in culture by inhibiting the RNAseH activity with an estimated EC₅₀ of ∼5 μM and a CC₅₀ of 10.1±1. 7 μM. Activity of β-thujaplicinol against RNAseH sequences from multiple HBV genotypes implies that if chemical derivatives of β-thujaplicinol with improved efficacy and reduced toxicity can be identified, they would have promise as anti-HBV agents.
乙型肝炎病毒(HBV)是一种嗜肝 DNA 病毒,通过逆转录复制。它慢性感染超过 3.5 亿人,每年导致约 100 万人死亡。治疗主要采用核苷(酸)类似物,这些药物通过病毒逆转录酶很好地抑制病毒 DNA 合成,但很少能治愈感染,因此需要额外的治疗方法。逆转录需要病毒核糖核酸酶 H(RNAseH)在病毒 RNA 被复制成 DNA 后将其破坏。我们最近生产了活性重组 HBV RNAseH,并证明 HIV RNAseH 拮抗剂可以以高频率抑制 HBV 酶。在这里,我们将这些结果扩展到β-雪松醇,一种抑制 HIV RNAseH 的羟基化三酮。β-雪松醇在生化测定中抑制来自 HBV 基因型 D 和 H 的 RNAseH,IC₅₀ 值分别为 5.9±0.7 和 2.3±1.7 μM。它通过抑制 RNAseH 活性来阻断 A 和 D 基因型 HBV 在培养中的复制,估计 EC₅₀ 约为 5 μM,CC₅₀ 为 10.1±1.7 μM。β-雪松醇对来自多种 HBV 基因型的 RNAseH 序列的活性表明,如果可以确定具有改善疗效和降低毒性的 β-雪松醇化学衍生物,它们将有望成为抗 HBV 药物。