HBV Capsid Effectors

项目摘要/摘要 尽管有有效的疫苗可用，但流行病学数据估计全球约有20亿人 感染了乙肝病毒(乙肝病毒)。约有3.5亿人是慢性乙肝病毒携带者，而且处于高发期 有患肝炎、肝硬变和肝细胞癌的风险。当前的抗乙肝治疗选择 抑制病毒但不能治愈，需要昂贵的终生治疗。因此，发现和发展小说 治疗方法不仅要抑制病毒复制，而且要消除乙肝病毒感染，这是关键。当前 已批准的治疗方法未能靶向乙肝病毒共价闭合环状DNA(cccDNA；相关 病毒持续存在)或病毒衣壳，这是病毒增殖所必需的。我们的创新方法旨在 衣壳组装是复制所必需的，因为从cccDNA合成DNA只发生在 衣壳编码的粒子。病毒核心蛋白(CP)构成病毒衣壳组装和衣壳的亚基 组装调节剂(CAM)加速衣壳组装的动力学，从而防止Poll-pgRNA 复杂的包埋和阻断乙肝病毒的复制。CAM还干扰cccDNA转录/从头开始 在感染的早期阶段形成。作为我们正在进行的HBVCAM发现计划的一部分，NIH支持 在过去的5年里，我们已经成功地开发了几个功能强大的II类凸轮，并取得了领先地位。 该化合物将于2020年10月进入1期临床试验。为了推进能量场并有后备化合物 通过改进的轮廓，我们最近鉴定了几个CAM同源和异源二聚体衍生物，显示出抗- 培养物中的乙肝病毒活性在皮摩尔范围内。因为连接两个CAM部分的二聚体结构可以相互作用 对于一个衣壳的两个截然不同的位置或最终将两个衣壳连接在一起，我们假设这些 与已知的I类或II类凸轮相比，化合物对乙肝病毒衣壳组装的影响更深远。基座 关于这些化合物(我们称之为III类)的效力和独特的作用模式，我们建议 通过追求三个具体目标来评估它们：1)在化学上优化和表征一系列独特的 2)从结构上、生化和生物学上表征新的CAM高分子 以及异源二聚体与乙肝病毒衣壳的相互作用；3)测定其药代动力学和体内药效 新型CAM同源和异源二聚体。将合成和评估新的同源和异源二聚体以达到 最大效力和类似药物的特性。为了将我们的化合物与现有的I类和II类凸轮区分开来， 我们将通过确定a)新凸轮的结构和动力学效应来表征它们对 乙肝病毒衣壳的形态及其在细胞内的定位，b)与乙肝病毒野生型和已知突变株CP结合， C)对主要的对CAM耐药的乙肝病毒株的耐药性和活性；d)衣壳内或衣壳间 关系。拟议研究的结果将验证我们的新型CAM类别，当与 其他方法可以为消除乙肝病毒的新治疗策略提供临床前的概念证明 同时缩短治疗时间，并避免进展为其他并发症。

PROJECT SUMMARY/ABSTRACT Despite the availability of an effective vaccine, epidemiologic data estimates about 2 billion people globally are infected with hepatitis B virus (HBV). Approximately 350 million people are chronic HBV carriers and at high risk for the development of hepatitis, cirrhosis and hepatocellular carcinoma. Current anti-HBV treatment options suppress the virus but do not cure, requiring costly lifetime therapy. Thus, discovering and developing novel therapeutic approaches to not only suppress viral replication, but also eliminate HBV infection is key. Current approved therapeutic approaches fail to target the HBV covalently closed-circular DNA (cccDNA; associated with viral persistence) or the virus capsid which is essential for virus proliferation. Our innovative approach targets capsid assembly which is essential for replication, as DNA synthesis from cccDNA occurs exclusively within the capsid encoded particle. HBV core proteins (Cp) constitute the subunits in viral capsid assembly and Capsid Assembly Modulators (CAM) accelerate the kinetics of capsid assembly whereby they prevent pol-pgRNA complex encapsidation and block HBV replication. CAMs also interfere with cccDNA transcription/de novo formation during early steps of infection. As part of our ongoing HBV CAM discovery program NIH-supported over the last 5 years, we have been successful in developing several highly potent class II CAMs with one lead compound entering phase 1 clinical trials in October 2020. To advance the field and have back up compounds with improved profile, we recently identified several CAM homo- and hetero-dimer derivatives displaying anti- HBV activity in culture in the picomolar range. Because a dimeric structure linking two CAM moieties can interact with two distinct sites of one capsid or eventually connect two capsids together, we hypothesized that these compounds would have a more profound impact on HBV capsid assembly than known class I or II CAMs. Based on the potency and the unique mode of action of these compounds (which we call Class III), we propose to evaluate them by pursuing three specific aims: 1) To chemically optimize and characterize a unique series of CAM homo and heterodimers; 2) To structurally, biochemically, and biologically characterize novel CAM homo and heterodimers binding interaction with HBV capsid; 3) To determine pharmacokinetics and in vivo efficacy of novel CAM homo and heterodimers. Novel homo and heterodimers will be synthesized and evaluated to reach maximum potency and drug-like properties. To differentiate our compounds from existing class I and II CAMs, we will characterize structural and dynamical effects of our new CAMs by determining a) their effect on the morphology of HBV capsids and their localization within cells, b) binding to HBV wild type and known mutant Cp, c) resistance profile and activity against major CAM resistant HBV strains and d) intra- or inter-capsid connections. Results from the proposed studies will validate our novel class of CAM, which, when combined with other modalities could provide preclinical proof of concept towards a novel therapeutic strategy to eliminate HBV while reducing treatment duration and also eliminate progression to other complications.