Vif Antagonism: Lead Discovery and SAR - Project 1

Vif 拮抗：先导化合物发现和 SAR - 项目 1

• 批准号: 8723303
• 负责人: TARIQ M RANA
• 金额: $ 40.88万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8723303
• 关键词: AIDS therapy; Acquired Immunodeficiency Syndrome; Animals; Antiviral Agents; Biological Assay; Cells; Clinic; Clinical; Collaborations; Complex; Cytidine; Deamination; Degradation Pathway; Development; Enzymes; Evaluation; Goals; HIV; HIV-1; Homologous Protein; Immune; Infection; Inhibitory Concentration 50; Instruction; Intervention; Lead; Macaca; Measures; Mediating; Medical; Molecular; Natural Immunity; Penetration; Peptide Hydrolases; Pharmaceutical Preparations; Pharmacology; Plasma; Property; RNA-Directed DNA Polymerase; Retroviridae; Safety; Structure; Structure-Activity Relationship; Testing; Therapeutic; Toxic effect; Validation; Viral; Viral Genome; Virion; Virus; analog; base; design; genetic regulatory protein; high throughput screening; human DNA; improved; in vivo; inhibitor/antagonist; innovation; interdisciplinary approach; multicatalytic endopeptidase complex; novel; novel strategies; pre-clinical; protein degradation; protein expression; research study; scaffold; small molecule; success; validation studies; vif Gene Products; viral resistance

PROJECT SUMMARY (See instructions): Our goal is to develop inhibitors against a novel and unexploited target for the treatment of AIDS. The human immunodeficiency virus type 1 (HlV-1), the causative agent of AIDS, is a complex retrovirus that encodes six regulatory proteins, including Vif that is essential for viral replication in vivo. Despite remarkable medical advances, HlV-1 infections continue to rise throughout the world. Current anti-HIV-1 agents target mainly HlV-1 reverse transcriptase or protease. However, inhibitors used to target these enzymes have given rise to viral resistance and related toxic effects, creating a need for more potent and less toxic therapies against other viral targets. HIV-1 is a complex retrovirus that encodes 6 regulatory proteins, among which Vif is essential for in vivo viral replication. HIV-1 Vif protein targets an innate antiviral human DNA-editing enzyme, AP0BEC3G (A3G), which inhibits replication of retroviruses. Since HiV-1 Vif has no known cellular homologs, this protein represents an extremely attractive; yet unrealized, target for antiviral intervention. Over the last few years, in collaboration with Dr. Stevenson (Project 2) and Core B, we developed and employed high throughput screening (HTS) assays to identify small molecules that antagonize HIV-1 Vif function. We focused our efforts to characterize one of these lead molecules, RN-18, which inhibits HlV-1 replication only in the presence of A3G. Mechanistic studies revealed that RN-18 enhanced Vif degradation only in the presence of A3G, increased A3G incorporation into virions leading to less infectious viruses, and enhanced cytidine deamination of the viral genome. These studies provide the first evidence that the HIV-1 Vif-A3G axis is a valid target for developing small molecule-based new therapies for AIDS or for enhancing innate immunity against viruses. Based on our preliminary success to develop Vif antagonists as a new class of AIDS therapeutics, we plan to employ highly innovative, collaborative, and multidisciplinary approaches to accomplish our proposed goals. Our project has the following specific aims: Specific Aim 1: Structure activity relationship studies and validation of lead compounds. Specific Aim 2: Structure activity relationship studies, lead optimization, and validation of unexplored scaffolds of Vif antagonists. Specific Aim 3: (i) Mechanism of Vif inhibition. Experiments will be performed to understand the mechanism of Vif inhibitors at molecular level, (ii) In vivo evaluation of Vif inhibitors. During the course of pharmacology studies (Project 3 and core B), we will measure the drug concentrations in plasma and CNS of drug treated animals.

项目总结（见说明）： 我们的目标是开发一种针对艾滋病治疗的新的和未开发的目标的抑制剂。人类免疫缺陷病毒1型（HIV-1）是AIDS的病原体，是一种复杂的逆转录病毒，其编码六种调节蛋白，包括对病毒在体内复制至关重要的Vif。尽管有显著的医学进步，但HIV-1感染在全世界继续上升。目前的抗HIV-1药物主要靶向HIV-1逆转录酶或蛋白酶。然而，用于靶向这些酶的抑制剂已经引起了病毒耐药性和相关的毒性作用，从而需要针对其他病毒靶点的更有效和毒性更小的疗法。HIV-1是一种复杂的逆转录病毒，编码6种调节蛋白，其中Vif是体内病毒复制所必需的。HIV-1 Vif蛋白靶向一种先天性抗病毒人类DNA编辑酶AP 0 BEC 3G（A3 G），可抑制逆转录病毒的复制。由于HIV-1 Vif没有已知的细胞同源物，这种蛋白代表了一个非常有吸引力的，但尚未实现的抗病毒干预的目标。在过去的几年里，我们与史蒂文森博士（项目2）和核心B合作，开发并采用高通量筛选（HTS）测定来鉴定拮抗HIV-1 Vif功能的小分子。我们集中精力来表征这些先导分子之一RN-18，其仅在A3 G存在下抑制HIV-1复制。机理研究表明，RN-18仅在A3 G存在下增强Vif降解，增加A3 G掺入病毒体中，导致感染性较低的病毒，并增强病毒基因组的胞苷脱氨基作用。这些研究提供了第一个证据，证明HIV-1 Vif-A3 G轴是开发基于小分子的艾滋病新疗法或增强对病毒的先天免疫的有效靶点。基于我们初步成功开发Vif拮抗剂作为一类新的艾滋病治疗药物，我们计划采用高度创新，协作和多学科的方法来实现我们提出的目标。我们的项目有以下具体目标：具体目标1：先导化合物的构效关系研究和验证。具体目标2：Vif拮抗剂的结构活性关系研究、先导物优化和未开发支架的验证。具体目标3：（i）Vif抑制机制。将进行实验以在分子水平上理解Vif抑制剂的机制。（ii）Vif抑制剂的体内评价。在药理学研究过程中（项目3和核心B），我们将测量给药动物血浆和CNS中的药物浓度。