Proteasomal Degradation of XPB as a Novel Mechanism for Treating Inflammation

XPB 的蛋白酶体降解作为治疗炎症的新机制

• 批准号: 10929222
• 负责人: Jason Elinoff
• 金额: $ 41.54万
• 依托单位: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10929222
• 关键词: Acceleration; Anti-Inflammatory Agents; Antiinflammatory Effect; Arteries; Autoimmune Diseases; Biological Assay; Cell model; Cessation of life; Chemicals; Clinic; Clinical; Clinical Trials; Collaborations; Complex; Disease; Dose; Drug Industry; Drug Kinetics; Endothelial Cells; Endothelium; Exclusion; Functional disorder; Gene Expression; Gene Silencing; Genes; Genetic Transcription; Heart failure; Human; In Vitro; Inflammation; Inflammatory; Informatics; Lead; Left; Lung; Mineralocorticoid Receptor; Molecular; National Center for Advancing Translational Sciences; Patients; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacologic Substance; Production; Proliferating; Proteins; Proteome; RNA interference screen; Reporter; Safety; Signal Pathway; Spironolactone; Techniques; Testing; Therapeutic; Translations; United States National Institutes of Health; Vascular Endothelium; Xeroderma Pigmentosum; antagonist; autoinflammatory diseases; clinical center; clinically relevant; cytokine; cytotoxic; drug candidate; drug development; functional genomics; genome-wide; high throughput screening; improved; innovation; lung vascular inflammation; multicatalytic endopeptidase complex; novel; novel therapeutics; pharmacologic; prototype; pulmonary arterial hypertension; pulmonary vascular cells; right ventricular failure; side effect; small molecule; small molecule libraries; transcription factor TFIIH; vascular inflammation

In collaboration with the NCATS Functional Genomics Lab, we have created and validated a high-throughput luminescent reporter assay suitable for identifying small molecule degraders of XPB and genome-wide RNAi screening. Using this state-of-the-art high-throughput assay we will: 1) Identify novel, small-molecule XPB degraders with potent anti-inflammatory effects that can be accelerated into clinical trials; and 2) Characterize molecular partners necessary for drug-induced XPB degradation in order to construct a more generalizable pharmaceutical paradigm. This innovative assay platform will enable discovery of clinically relevant XPB degraders that modulate lung vascular inflammation in PAH patients, and the NIH Clinical Center is ideally suited for spearheading trials testing these first-in-class anti-inflammatory drugs. Project Milestones: - Screen the NCATS Small Molecule Library (up to 150,000 compounds) for drugs that induce XPB-HiBiT degradation. Candidate drugs will be selected through bio/chemi-informatic analyses, validated at 11 concentrations, and undergo counter screens to exclude cytotoxic effects and/or assay interference. Top candidates will be resynthesized and their activity confirmed. - Perform a second, high-throughput screen of candidate XPB degraders for potent anti-inflammatory effects across heterogeneous signaling pathways. Novel drugs identified in both screens will be further examined for their effects on inflammatory gene transcription and cytokine production in cellular models of PAH-associated endothelial cell dysfunction. - Perform a genome-wide RNAi screen to define the molecular mechanism underlying drug-induced XPB degradation. Target deconvolution of candidate drugs that induce proteasome-dependent XPB degradation and the NCATS pharmacologically annotated chemical toolbox will be used to substantiate gene silencing effects. Candidate XPB degradation genes will be confirmed in primary human pulmonary vascular endothelium. - Apply state-of-the-art medicinal chemistry techniques to optimize pharmacokinetics, potency, and efficacy of lead anti-inflammatory drugs for translation into the clinic.

与NCATS功能基因组学实验室合作，我们创建并验证了一种高通量发光报告基因检测方法，适用于鉴定XPB的小分子降解物和全基因组RNAi筛选。 使用这种最先进的高通量检测方法，我们将： 1)鉴定具有强效抗炎作用的新型小分子XPB降解剂，其可以加速进入临床试验;和2）表征药物诱导的XPB降解所必需的分子伴侣，以构建更普遍的药物范例。这种创新的检测平台将能够发现调节PAH患者肺血管炎症的临床相关XPB降解剂，NIH临床中心非常适合率先试验这些一流的抗炎药物。 项目里程碑： - 筛选NCATS小分子库（多达150，000种化合物）中诱导XPB-HiBiT降解的药物。候选药物将通过生物/化学信息学分析进行选择，在11个浓度下进行验证，并进行计数器筛选以排除细胞毒性效应和/或测定干扰。最佳候选人将被重新合成，并确认他们的活动。 - 对候选XPB降解剂进行第二次高通量筛选，以获得跨异质信号通路的强效抗炎作用。在两种筛选中鉴定的新药将进一步检查其对PAH相关内皮细胞功能障碍细胞模型中炎症基因转录和细胞因子产生的影响。 - 进行全基因组RNAi筛选，以确定药物诱导XPB降解的分子机制。诱导蛋白酶体依赖性XPB降解的候选药物的靶向去卷积和NCATS注释的化学工具箱将用于证实基因沉默效应。候选XPB降解基因将在原代人肺血管内皮中得到确认。 - 应用最先进的药物化学技术来优化主要抗炎药物的药代动力学，效力和疗效，以转化为临床。