Vif Antagonism: Lead Discovery and SAR - Project 1

Vif 拮抗：先导化合物发现和 SAR - 项目 1

基本信息

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
项目类别：
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

项目摘要

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.

项目摘要（请参阅说明）：我们的目标是开发针对艾滋病治疗的新颖且未开发的目标的抑制剂。人类免疫缺陷病毒1型（HLV-1）是AIDS的致病药物，是一种复杂的逆转录病毒，编码六种调节蛋白，包括VIF，这对于体内病毒复制至关重要。尽管医疗进展显着，但HLV-1感染仍在全球范围内继续增加。当前的抗HIV-1药物主要靶向HLV-1逆转录酶或蛋白酶。然而，用于靶向这些酶的抑制剂引起了病毒抗性和相关的毒性作用，从而需要针对其他病毒靶标的更有效，更毒性的疗法。 HIV-1是一种复杂的逆转录病毒，编码6种调节蛋白，其中VIF对于体内病毒复制至关重要。 HIV-1 VIF蛋白靶向先天抗病毒人DNA编辑酶，AP0BEC3G（A3G），该酶抑制了逆转录病毒的复制。由于HIV-1 VIF没有已知的细胞同源物，因此该蛋白质代表着一种极具吸引力。尚未实现的抗病毒干预目标。在过去的几年中，与Stevenson博士（项目2）和Core B合作，我们开发并采用了高吞吐量筛选（HTS）测定法，以识别与HIV-1 VIF功能拮抗的小分子。我们集中精力表征其中一种铅分子RN-18，该分子仅在A3G存在下抑制HLV-1复制。机械研究表明，RN-18仅在A3G存在下才能增强VIF降解，增加了A3G掺入病毒体中，导致感染性较低的病毒，并增强了病毒基因组的胞苷脱氨酸。这些研究提供了第一个证据，即HIV-1 VIF-A3G轴是开发基于小分子的新艾滋病疗法或增强对病毒的先天免疫力的有效靶标。基于我们将VIF拮抗剂作为新的AIDS治疗师发展的初步成功，我们计划采用高度创新，协作和多学科的方法来实现我们建议的目标。我们的项目具有以下特定目的：特定目的1：结构活动关系研究和铅化合物的验证。特定目标2：结构活动关系研究，铅优化以及对VIF拮抗剂未开发的支架的验证。特定目标3：（i）VIF抑制的机制。将进行实验以了解分子水平上VIF抑制剂的机制，（ii）VIF抑制剂的体内评估。在药理学研究过程中（项目3和核心B），我们将测量药物治疗动物的血浆和中枢神经系统中的药物浓度。