Discovery of novel anti-HBV compounds targeting host factors

发现针对宿主因子的新型抗乙肝病毒化合物

• 批准号: 8731770
• 负责人: Alan McLachlan
• 金额: $ 19.76万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-09 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8731770
• 关键词: Adherence; Anabolism; Antiviral Agents; Antiviral Therapy; Biological Assay; Biological Factors; Biology; Carrier State; Cell Culture Techniques; Cells; Chemicals; Chicago; Chronic; Chronic Hepatitis B; Clinic; Communities; Complement; Computer Assisted; Consensus; Development; Drug Design; Drug Kinetics; Drug Targeting; Evaluation; Generations; Genetic Transcription; Goals; Hepatitis B Virus; Illinois; Integration Host Factors; Interferons; Lead; Libraries; Life; Life Cycle Stages; Mass Spectrum Analysis; Mediating; Methods; Molecular; Molecular Models; Molecular Weight; Nuclear Receptors; Nucleocapsid; Patients; Pharmaceutical Chemistry; Pharmaceutical Preparations; Photoaffinity Labels; Prevention; Probability; RNA chemical synthesis; RNA-Directed DNA Polymerase; Records; Research; Research Personnel; Resolution; Techniques; Therapeutic; Therapeutic Agents; Time; Toxic effect; Toxicology; Universities; Vial device; Viral; Virus Diseases; Work; analog; animal efficacy; anti-hepatitis B; assay development; base; cellular targeting; design; efficacy evaluation; experience; hepatitis B virus P protein; high throughput screening; inhibitor/antagonist; molecular modeling; novel; novel strategies; novel therapeutics; nucleoside analog; promoter; public health relevance; research study; scaffold; screening; small molecule; small molecule libraries; therapy development

DESCRIPTION (provided by applicant): This R21 exploratory proposal uses an integrated approach to discover lead candidates suitable for further lead optimization as therapeutics for chronic hepatitis B virus (HBV) infections. Our long-term objective is to develop small molecule inhibitors of HBV biosynthesis with a novel mechanism of action distinct from the currently available drugs. A broad consensus exists in the HBV research community that the key to the successful development of new novel classes of HBV medications lies in targeting steps in the viral life cycle other than replication mediated by the HBV polymerase/reverse transcriptase. Currently available medications, including type 1 interferon and nucleosides analogs, do not offer a significant probability for the resolution of the chronic carrier state. The shortcomings o the existing HBV therapeutic regiments include the necessity of life-long treatment and suboptimal patient acceptance and adherence. Clearly, new drugs with novel mechanisms of action are needed to find a cure leading to complete eradication of HBV infection. Antiviral therapy development targeting not viral, but cellular components essential for viral biosynthesis may represent a novel approach for the generation of HBV therapeutics. A combination of nucleoside analogs with the novel drugs targeting host factors may potentially lead to the prevention of new cccDNA synthesis. Recent advances in target identification techniques for therapeutically relevant chemical compounds and the availability of simple assays amenable to HTS analysis for the evaluation of HBV transcription allow for a multitude of methods to discover new therapeutic agents for HBV. We hypothesize that it will be possible to identify and design inhibitors of HBV biosynthesis by specifically targeting cellular factors that are essential for vial pregenomic RNA synthesis using a combination of approaches: high- throughput screening (HTS) against HBV nucleocapsid promoter activity, computer-aided drug design (CADD), chemical biology, target identification methods, target-oriented rational drug design, medicinal chemistry, and evaluation for efficacy of the most promising compounds shown to inhibit HBV transcription and replication in cell culture. To achieve this goal, our specific aims are as follo: Aim 1. Identify lead- or drug-like low molecular weight inhibitors of HBV nucleocapsid promoter activity using high- throughput screening of chemical libraries with available primary HTS, counter-screening HTS, and secondary viral replication assays. Aim 2. Design and synthesize analogs of the most promising HBV antiviral compounds from aim 1 to generate a SAR and identify compounds suitable for cell-based and photoaffinity-based target identification experiments. If time permits, pilot target identification studies using non-hepatoma cells complemented with a panel of nuclear receptors capable of supporting HBV transcription and replication, and photoaffinity labeling followed by LC-MS/MS analysis will be performed.

描述（由申请人提供）：该 R21 探索性提案使用综合方法来发现适合进一步优化作为慢性乙型肝炎病毒 (HBV) 感染治疗方法的先导候选药物。我们的长期目标是开发具有不同于现有药物的新颖作用机制的 HBV 生物合成小分子抑制剂。 HBV 研究界存在广泛共识，即成功开发新型 HBV 药物的关键在于针对病毒生命周期中的步骤，而不是 HBV 聚合酶/逆转录酶介导的复制。目前可用的药物，包括 1 型干扰素和核苷类似物，对于解决慢性携带状态没有提供显着的可能性。现有乙肝治疗方案的缺点包括需要终身治疗以及患者接受度和依从性不佳。显然，需要具有新颖作用机制的新药来找到彻底根除乙型肝炎病毒感染的治疗方法。抗病毒疗法的开发不是针对病毒，而是针对病毒生物合成所必需的细胞成分，这可能代表了一种产生乙型肝炎疗法的新方法。核苷类似物与针对宿主因子的新药物的组合可能会阻止新的 cccDNA 合成。治疗相关化合物的靶标识别技术的最新进展以及适用于 HTS 分析的简单测定法的可用性以评估 HBV 转录，允许采用多种方法来发现新的 HBV 治疗剂。我们假设，通过专门针对小瓶前基因组 RNA 合成所必需的细胞因子，可以使用以下方法组合来识别和设计 HBV 生物合成抑制剂：针对 HBV 核衣壳启动子活性的高通量筛选 (HTS)、计算机辅助药物设计（CADD）、化学生物学、靶点识别方法、以靶点为导向的合理药物设计、药物化学以及对最有希望抑制细胞培养中 HBV 转录和复制的化合物的功效进行评估。为了实现这一目标，我们的具体目标如下： 目标 1. 使用可用的初级 HTS、反筛选 HTS 和反筛选 HTS 来高通量筛选化学文库，鉴定 HBV 核衣壳启动子活性的先导或药物样低分子量抑制剂。二次病毒复制测定。目标 2. 设计和合成目标 1 中最有前途的 HBV 抗病毒化合物的类似物，以生成 SAR 并识别适合基于细胞和基于光亲和力的目标识别实验的化合物。如果时间允许，将使用非肝癌细胞进行试点目标识别研究，这些细胞辅以一组能够支持 HBV 转录和复制的核受体，以及光亲和标记，然后进行 LC-MS/MS 分析。