Specific activation of latent HIV-1 by inhibiting E3 ubiquitin ligase activity

通过抑制 E3 泛素连接酶活性特异性激活潜伏 HIV-1

基本信息

批准号：
9197394
负责人：
JOSEPH DOUGHERTY
金额：
$ 40.25万
依托单位：
RBHS-ROBERT WOOD JOHNSON MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-11-15 至 2019-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9197394
关键词：
Affect Bioavailable Biological Assay Bone Marrow Cell Culture System Cells Chemicals Collection Combined Modality Therapy Complex Development Drug Interactions Drug Kinetics Epigenetic Process Gene Expression Gene Expression Regulation Generations Genetic Transcription Glean Goals HIV-1 Health Highly Active Antiretroviral Therapy Human Immunotherapy Individual Infection Latent Virus Lead Libraries Life Literature Liver Maintenance Modeling Modification Molecular Weight Mus Patients Pattern Pharmaceutical Chemistry Pharmaceutical Preparations Pharmacologic Substance Phase Play Property Proteasome Inhibition Proteasome Inhibitor Quantitative Structure-Activity Relationship RNA Polymerase II RNA interference screen Regulation Reporting Role Safety Source Specificity Structure System Techniques Testing Therapeutic Thymus Gland Toxic effect Ubiquitin Validation Virus Virus Latency analog arm base chromatin remodeling cytotoxicity design drug development efficacy testing genome-wide high throughput screening improved in vivo insight latent virus activation memory CD4 T lymphocyte multicatalytic endopeptidase complex pharmacophore protein degradation research study scaffold small molecule small molecule inhibitor transcription factor ubiquitin-protein ligase

项目摘要

DESCRIPTION (provided by applicant): HIV-1 persists indefinitely in infected individuals including those successfully being treated with highly active antiretroviral therapy (HAART). Numerous studies indicate that latently infected long-lived memory CD4+ T cells are an important source of enduring infection. A strategy that is being actively pursued to eliminate the latent arm of the infection involves developing and employing drugs that act as antagonists of viral latency. To date, the latency antagonists that have been identified tend to lack a high degree of specificity. Moreover, the mechanism(s) underlying viral latency in patients is likely multifactorial. Therefore, it may be necessary to develop combination therapies possibly in conjunction with immunotherapy to ultimately eliminate the virus from patients. Drugs, which can activate latent virus with high specificity are likely to have fewer off-target effects and might b significantly less toxic in the context of combination therapy. The ubiquitin-proteasome system (UPS) has been shown to play an important role in gene regulation. A great deal of specificity is imparted to the UPS particularly by the E3 ubiquitin ligases, of which there are more than 600 in human cells. Preliminary studies indicate that the UPS is involved in maintaining HIV-1 latency since proteasome inhibitors (PIs) can activate latent virus. This has led to the hypothesis that there is a factor positively affecting HIV-1 gene expression that is targeted for proteasomal degradation in latently infected cells. Therefore, it should be possible to identify the factor and the specific E3 ligase that directs it turnover, leading to development of drugs that can specifically inhibit ubiquitylation of the positive factor activating latent virus. The R21 phase (Aims 1 and 2) of this proposal aims at identifying the positive factor and the specific E3 ligase targeting its turnover. Upon successful completion of the R21, the R33 phase (Aims 3 and 4) would involve: 1) developing an assay suitable for high-throughput screening (HTS), 2) carrying out a HTS with a library of over 200,000 diversified low molecular weight compounds, 3) characterizing hits and selecting lead compounds, and 4) conducting lead optimization to improve the potency, selectivity, and pharmaceutical properties of lead compounds, aimed at the identification of an orally available Development Candidate.

描述（由申请人提供）：HIV-1 在感染个体中无限期持续存在，包括那些成功接受高效抗逆转录病毒疗法 (HAART) 治疗的个体。大量研究表明，潜伏感染的长寿命记忆 CD4+ T 细胞是持久感染的重要来源。目前正在积极推行消除潜伏感染的策略，包括开发和使用作为病毒潜伏拮抗剂的药物。迄今为止，已鉴定的潜伏拮抗剂往往缺乏高度的特异性。此外，患者病毒潜伏期的机制可能是多因素的。因此，可能有必要开发联合疗法，可能与免疫疗法相结合，以最终消除患者体内的病毒。能够以高特异性激活潜伏病毒的药物可能具有较少的脱靶效应，并且在联合治疗的情况下毒性可能显着降低。泛素蛋白酶体系统（UPS）已被证明在基因调控中发挥重要作用。 UPS 具有很大的特异性，尤其是 E3 泛素连接酶，人类细胞中有 600 多种泛素连接酶。初步研究表明，UPS 参与维持 HIV-1 潜伏期，因为蛋白酶体抑制剂 (PI) 可以激活潜伏病毒。这导致了这样的假设：存在一个对 HIV-1 基因表达产生积极影响的因素，该因素针对潜伏感染细胞中的蛋白酶体降解。因此，应该能够识别该因素并指导其转换的特定 E3 连接酶，导致开发出能够特异性抑制激活潜伏病毒的阳性因子泛素化的药物。该提案的 R21 阶段（目标 1 和 2）旨在确定积极因素和针对其营业额的特定 E3 连接酶。成功完成 R21 后，R33 阶段（目标 3 和 4）将涉及：1) 开发适合高通量筛选 (HTS) 的测定方法，2) 使用包含超过 200,000 个多样化低分子量的文库进行 HTS化合物，3) 表征命中并选择先导化合物，以及 4) 进行先导化合物优化以提高先导化合物的效力、选择性和药物特性，旨在鉴定口服可用的开发候选人。