Novel antivirals targeting the RNase H activity of HIV reverse transcriptase

针对 HIV 逆转录酶 RNase H 活性的新型抗病毒药物

• 批准号: 8419398
• 负责人: MICHAEL A PARNIAK
• 金额: $ 71.05万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8419398
• 关键词: Achievement; Active Sites; Address; Anti-Retroviral Agents; Antiviral Agents; Binding Sites; Biochemical; Biochemistry; Biological; Biological Assay; Cells; Complement; Complex; Computational Biology; Computer Analysis; Computer Assisted; Crystallization; DNA-Directed DNA Polymerase; Data; Databases; Development; Docking; Drug Design; Drug resistance; Enzymatic Biochemistry; Enzymes; Future; Genome; Goals; HIV; Highly Active Antiretroviral Therapy; Integrase; Laboratories; Lead; Mediating; Modification; Molecular; Molecular Models; Patients; Peptide Hydrolases; Pharmaceutical Chemistry; Pharmaceutical Preparations; Process; Property; Protocols documentation; RNA-Directed DNA Polymerase; Resistance; Resolution; Reverse Transcription; Ribonuclease H; Robotics; Screening procedure; Structure; Structure-Activity Relationship; System; Therapeutic; Validation; Variant; Viral Load result; analog; base; chemical synthesis; design; drug candidate; drug resistant virus; improved; inhibitor/antagonist; interdisciplinary approach; molecular modeling; novel; resistance mutation; resistant strain; scaffold; structural biology; tool; viral RNA; virology

DESCRIPTION (provided by applicant): Highly active antiretroviral therapy (HAART) drugs primarily target three of the four HIV enzymes: reverse transcriptase (RT) DNA polymerase, protease, and integrase. Although HAART is very effective in suppressing viral load in HIV-infected patients, prolonged treatment inevitably leads to the emergence of drug-resistant viral strains. Hence, it is essential to develop agents that act on novel HIV targets. The fourth HIV enzyme, RT- associated ribonuclease H (RNH) is one such target. RNH degrades the viral RNA genome during reverse transcription and is essential for HIV replication. It is the only enzymatic activity of HIV that has yet to be addressed by antiretroviral drugs. Such drugs will likely be active against all current drug-resistant viral strains. Our goal is to develop potent RNH inhibitors (RNHIs) with nanomolar efficacy in cell-based replication assays and for this we will use a multidisciplinary approach based on our extensive expertise in structural biology, computational biology, medicinal chemistry, enzymology, biochemistry and virology. We will pursue improvement of current leads and achievement of nM potencies of antiviral activities through a structure-based design process involving iterative cycles of structure determination and computational analysis of RNH-RNHI complexes, medicinal chemistry, and biochemical and virological characterization of newly synthesized inhibitors. To this end we propose three specific aims: Specific Aim 1. Structure-Activity Relationships (SAR) and Chemical Synthesis. We will prepare a database of the validated RNHI screening hits and novel scaffolds that we have developed. We will perform complete SAR for two different classes of RNHIs based on the database analysis and the structural information gained from crystallographic and molecular docking studies in aim 2. Specific Aim 2. Crystallographic and computational analysis of RT-RNHI interaction. Structure-based design is a main focus of this application. We will use crystallographic tools that are already established in our lab and that routinely result in high resolution structures of RT and/or RNH in complex with inhibitors (resolutions up to 1.5 ¿). The structural information will be used to guide the design of new inhibitors. Specific Aim 3. Biochemical and virologic profiling of RNHIs. We will use biochemical and virological assays to assess selected validated screening hits and new RNHIs to be prepared in Aim 1. This information will be integrated in the iterative SAR-mediated design of new inhibitors. Our multidisciplinary approach will lead to new inhibitors of HIV that will be effective against both wild- type and drug-resistant viral strains. PUBLIC HEALTH RELEVANCE: This project will discover compounds that target an essential biological activity of the HIV virus, that currently approved drugs do not address. These studies should lead to the development of future therapeutics that complement existing drugs and should also strengthen our efforts to block drug- resistant strains of HIV.

描述（由申请人提供）：高效抗逆转录病毒疗法（HAART）药物主要针对四种 HIV 酶中的三种：逆转录酶（RT）DNA 聚合酶、蛋白酶和整合酶，尽管 HAART 在抑制 HIV 感染者的病毒载量方面非常有效。对于患者来说，长期治疗不可避免地会导致耐药病毒株的出现，因此，开发针对第四种 HIV 酶（RT 相关酶）的药物至关重要。核糖核酸酶 H (RNH) 就是这样的靶标之一，RNH 在逆转录过程中降解病毒 RNA 基因组，并且是 HIV 复制所必需的，它是抗逆转录病毒药物可能具有活性的唯一酶活性。我们的目标是开发在基于细胞的复制测定中具有纳摩尔功效的强效 RNH 抑制剂 (RNHI)，为此，我们将使用基于我们在结构生物学方面的广泛专业知识的多学科方法，我们将通过基于结构的设计过程（涉及 RNH-RNHI 复合物的结构确定和计算分析的迭代循环）来改进当前的线索并实现 nM 的抗病毒活性。新合成抑制剂的化学、生化和病毒学表征 为此，我们提出了三个具体目标： 具体目标 1. 结构-活性关系 (SAR) 和化学。合成。我们将准备一个经过验证的 RNHI 筛选命中和我们开发的新型支架的数据库。我们将根据数据库分析以及从晶体学和分子对接研究中获得的结构信息，对两类不同的 RNHI 进行完整的 SAR。 2. 具体目标 2. 基于结构的设计的晶体学和计算分析是本应用的主要焦点，我们将使用我们实验室已经建立的晶体学工具，并且常规结果是高分辨率。 RT 和/或 RNH 与抑制剂复合物的结构（分辨率高达 1.5 ¿ ). 结构信息将用于指导新抑制剂的设计。具体目标 3. RNHI 的生化和病毒学分析。我们将使用生化和病毒学分析来评估目标 1 中选定的经过验证的筛选结果和新的 RNHI。这些信息将被整合到新抑制剂的迭代 SAR 介导设计中，我们的多学科方法将产生新的 HIV 抑制剂，对野生型和耐药病毒均有效。菌株。 公共健康相关性：该项目将发现针对 HIV 病毒的基本生物活性的化合物，而目前批准的药物无法解决这一问题。这些研究应导致开发出补充现有药物的未来疗法，并应加强我们的阻止努力。 HIV 的耐药菌株。