Optimization of alpha-hydroxytropolones as novel inhibitors of the HBV RNaseH

α-羟基托酚酮作为 HBV RNaseH 新型抑制剂的优化

• 批准号: 9390039
• 负责人: JOHN E TAVIS
• 金额: $ 44.01万
• 依托单位: SAINT LOUIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-12-01 至 2020-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9390039
• 关键词: Active Sites; Antiviral Agents; Binding; Biochemical; Biological Assay; Capsid; Cell Nucleus; Cells; Chemicals; Chronic; Clinical; DNA; DNA Viruses; DNA biosynthesis; DNA-Directed DNA Polymerase; Data; Detection; Development; Distant; Drug Design; Drug Kinetics; Drug Screening; Enzymes; Excretory function; Family; Future; Genome; Genomics; Goals; HIV; Hepatitis B Virus; Infection; Lead; Life; Maintenance; Measures; Metabolism; Nuclear; Pathway interactions; Patients; Pharmaceutical Chemistry; Pharmaceutical Preparations; RNA; RNA-Directed DNA Polymerase; Recombinants; Resistance; Reverse Transcription; Ribonuclease H; Structure-Activity Relationship; Testing; Therapeutic Index; Time; Toxic effect; Tropolone; Viral; Viral Genome; Virion; Virus; Virus Replication; Work; absorption; analog; anti-hepatitis B; base; cytotoxicity; design; drug development; drug discovery; inhibitor/antagonist; iterative design; mouse model; novel; novel therapeutics; nucleoside analog; prevent; public health relevance; screening; targeted treatment; viral DNA; viral RNA

 DESCRIPTION (provided by applicant): Hepatitis B virus (HBV) is a hepatotropic virus that replicates by reverse transcription. It chronically infects ~350 million people and kills >600,000 annually. Therapy primarily employs nucleos (t)ide analogs that drive HBV to below the limit of detection in most patients. However, viral replication is not eradicated and viral replication resurges if drug is withdrawn, so therapy is life-long. Nevertheless, current treatment does clear HBV in some patients, so more patients could be cured by suppressing HBV further. Greater suppression of HBV will require new drugs that will probably be used in combination with the nucleos (t)ide analogs. HBV reverse transcription requires a viral DNA polymerase to synthesize new DNA and a viral ribonuclease H (RNaseH) to destroy the RNA after its conversion into DNA. Blocking either activity prevents HBV replication. Drugs against the RNaseH do not exist due to an inability to produce enzyme suitable for screening. We recently produced HBV RNaseH suitable for drug discovery. We screened 54 troponoid compounds, focusing on the α-hydroxytropolones (αHTs), based on their ability to inhibit the HIV RNaseH and β-thujaplicinol's inhibition of the HBV RNaseH. Thirteen compounds inhibited the HBV RNaseH, and the best had an IC50 value of 2.3 µM. six compounds suppressed HBV replication in culture via blocking viral RNaseH activity. The best compound had an EC50 of 0.34 µM and a therapeutic index of 94. A preliminary structure-activity relationship (SAR) was derived that provides guidance for optimization by medicinal chemistry. Here, Dr. Tavis (virologist) and Drs. Murelli and Meyers (synthetic and medicinal chemists) will collaborate to begin development of the αHTs into potent anti-HBV drugs. Aim 1. Produce optimal αHT inhibitors of HBV replication. We will synthesize ~320 αHTs based on our preliminary SAR. These compounds will be tested in an iterative fashion for anti-HBV RNaseH activity in biochemical, viral replication, and cytotoxicity assays. Aim 2. Optimize cytotoxicity, ADME, PK, and efficacy parameters in a mouse model of HBV replication. Absorption, distribution, metabolism, excretion (ADME), initial pharmacokinetics (PK), and toxicity will be measured to guide compound design. Efficacy will be characterized in a mouse model of HBV replication for the leading inhibitors. Aim 3. Evaluate the potential for use of αHT compounds in combination with nucleos(t)ide analogs. We will assess efficacy of the optimized αHT compounds against common nucleos(t)ide analog- resistant HBV strains. We will also evaluate whether αHT inhibitors work synergistically with the nucleos(t)ide analogs as predicted from their binding to physically distinct active sites on the HBV reverse transcriptase. This project will produce the first αHT inhibitors designed against the HBV RNaseH and will define their potential to contribute to anti-HBV therapy. It may also produce lead compounds for drug development.

 描述（由适用提供）：丙型肝炎病毒（HBV）是一种通过逆转录复制的肝病病毒。它长期感染了约3.5亿人，每年杀死> 60万人。在大多数患者中，治疗初级雇员核能（T）IDE类似物，将HBV降低到检测极限。但是，病毒复制不是放射性的，如果撤出药物，则可以复制病毒复制，因此治疗是终身的。然而，当前的治疗确实清楚了某些患者的HBV，因此可以通过进一步抑制HBV来治愈更多的患者。对HBV的更大抑制作用将需要新药物，这些药物可能与核（T）IDE类似物结合使用。 HBV逆转录需要病毒DNA聚合酶来合成新的DNA和病毒核糖核酸酶H（RNASEH），以破坏RNA转换为DNA。阻止任何一种活动都阻止了HBV复制。由于无法产生适合筛查的酶，因此不存在针对RNASEH的药物。我们最近生产的HBV RNASEH适用于药物发现。我们根据抑制HIV RNASEH和β-thujaplicinol对HBV RNAseH的抑制作用的能力，筛选了54种肌红花化合物，重点介绍了α-羟基滋养体（αHTS）。 13种化合物抑制了HBV RNASEH，最好的IC50值为2.3 µm。六种化合物通过阻断病毒RNASEH活性抑制了培养中的HBV复制。最佳化合物的EC50为0.34 µm，治疗指数为94。得出了初步的结构活性关系（SAR），该关系提供了医学化学优化的指导。塔维斯博士（病毒学家）和博士。 Murelli和Meyers（合成和医学化学家）将合作，开始将αHT开发为潜在的抗HBV药物。 AIM 1。产生最佳的HBV复制抑制剂。我们将根据初步SAR合成〜320αHT。这些化合物将在生化，病毒复制和细胞毒性测定中以迭代方式进行抗HBV RNASEH活性进行测试。 AIM 2。在HBV复制的小鼠模型中优化细胞毒性，ADME，PK和效率参数。吸收，分布，代谢，排泄（ADME），初始药代动力学（PK）和毒性将测量以指导化合物设计。功效将在HBV复制的小鼠模型中为领先的抑制剂进行表征。 AIM 3。评估使用αHT化合物与核（T）IDE类似物结合使用的潜力。我们将评估针对常见核（T）IDE类似物HBV菌株的优化αHT化合物的效率。我们还将评估αHT抑制剂是否与核（t）IDE类似物协同工作，从它们与HBV逆转录酶上物理不同活性位点的结合预测。该项目将产生针对HBV RNASEH设计的第一个αHT抑制剂，并将定义其有助于抗HBV疗法的潜力。它也可能产生用于药物开发的铅化合物。