Development of in vivo active small molecule selective inhibitors of ASK1

ASK1 体内活性小分子选择性抑制剂的开发

基本信息

批准号：
9218535
负责人：
Derek Ronald Duckett
金额：
$ 59.83万
依托单位：
SCRIPPS FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-12-15 至 2019-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9218535
关键词：
Acute Adverse effects Affinity Animal Model Apoptosis Apoptosis Inhibitor Binding Proteins Biochemical Biochemistry Biological Biological Assay Biology Blood - brain barrier anatomy Brain Cardiovascular Diseases Cardiovascular system Cells Cellular Stress Cellular biology Chemicals Chemistry Clinical Complex Core Facility Critical Pathways Development Disease Dissociation Dose Drug Kinetics Enzymes Family Feedback Florida Genetic study Goals Heart Hypertrophy Histopathology In Situ Nick-End Labeling In Vitro Inflammatory Inhibition of Apoptosis Kinetics Knockout Mice Lead Legal patent Literature MAP2K6 gene MAPK14 gene MAPK8 gene Measurement Measures Mitogen-Activated Protein Kinases Molecular Probes Mus Neurodegenerative Disorders Organ P-Glycoprotein Performance Peripheral Pharmaceutical Chemistry Pharmaceutical Preparations Pharmacodynamics Pharmacology Phase Phenotype Phosphorylation Phosphotransferases Physiological Processes Play Property Purines Pyrimidines Reaction Regulation Reporter Reporting Research Research Infrastructure Research Proposals Rheumatoid Arthritis Role Safety Scanning Series Signal Transduction Solubility Staining method Stains Stress Structure Toxic effect Triage Triazines United States Food and Drug Administration Validation X-Ray Crystallography base chemical property design drug metabolism experimental study human disease improved in vivo insight kinase inhibitor member metabolic abnormality assessment mouse model nanomolar physical property pre-clinical programs pyridine receptor response scaffold screening small molecule small molecule inhibitor structural biology therapeutic target

项目摘要

Altered regulation of signaling in response to persistent cellular stress is associated with a number of complex human diseases, including cardiovascular, inflammatory and neurodegenerative disorders. Importantly, genetic studies have shown that silencing of the apoptosis signal-regulating kinase (ASK1) ameliorates many of the phenotypes manifest in mouse models of rheumatoid arthritis, cardiac hypertrophy and neurodegenerative disorders. Consequently, ASK1 has been highlighted as a therapeutic target where antagonists are anticipated to provide significant benefit in multiple disease states. Under the auspices of GM103957 our research team has recently completed a high throughput ASK1 screening campaign and identified a series of small molecule inhibitors from six different structural classes. We have narrowed the focus of our medicinal chemistry efforts to three scaffolds and the goal of this proposal is to develop an in vivo active ASK1 small molecule probe. In the studies of Aim 1 we will use an iterative medicinal chemistry approach to optimize the potency, selectivity and in vivo properties of ASK1 inhibitors aimed at peripheral targets. In addition, to develop compounds with suitable brain penetrant properties we will apply a multi-parameter optimization strategy focusing on physical and chemical properties that are hallmarks of the majority of current Food and Drug Administration (FDA)- approved CNS penetrant small molecule drugs. We will use structure based design to facilitate chemistry strategies and we will triage compounds through a battery of biochemical and cell-based assays as outlined in our research operating plan (ROP). As improved leads emerge, we will obtain enzyme selectivity for a select few, through full scale kinome (Reaction Biology) and receptor profiling (Riserca). For key leads we will also assess target engagement lifetime through biochemical and cell-based studies through measurement of ASK1 downstream substrate phosphorylation. Integrated into the ROP, we will also triage new compounds through a series of drug metabolism studies including assessment of in vitro microsomal stability, CYP450 inhibition, solubility, protein binding and P-glycoprotein efflux, with lead criteria integrated into the ROP for compound progression. In Aim 2, advanced leads for both peripheral and CNS exposures will be assessed in mice for adverse effects in acute dose range finding and 28-day toxicity studies. We will measure gross toxicity effect and vet scan analyses as well as TUNEL staining and histopathology of major organs. Finally, we will use inhibition of MKK6, JNK and p38 phosphorylation as in vivo reporters of ASK1 inhibition to quantify the maximum biological effective dose and through kinetic studies establish the PK/PD relationships of our lead in vivo probes. Our Multi-PI research team at the Scripps Florida campus of TSRI has both the expertise and infrastructure to perform these experiments, and we submit that the successful completion of our studies will identify a series of potent, selective and in vivo active molecular probes that will help define the role(s) that ASK1 plays in normal physiological processes as well as in disease states.

响应持续细胞应激而改变的信号传导调节与许多复杂的人类疾病，包括心血管疾病、炎症和神经退行性疾病。重要的是遗传研究表明，凋亡信号调节激酶 (ASK1) 的沉默可以改善许多表型在类风湿性关节炎、心脏肥大和神经退行性疾病的小鼠模型中表现出来失调。因此，ASK1 已被强调为预期有拮抗剂的治疗靶点在多种疾病状态下提供显着的益处。在GM103957的资助下我们的研究团队最近完成了高通量 ASK1 筛选活动并鉴定了一系列小分子来自六种不同结构类别的抑制剂。我们已将药物化学工作的重点缩小到三个支架，本提案的目标是开发体内活性 ASK1 小分子探针。在在目标 1 的研究中，我们将使用迭代药物化学方法来优化效力、选择性和针对外周靶标的 ASK1 抑制剂的体内特性。此外，开发化合物合适的大脑渗透特性，我们将应用专注于物理的多参数优化策略和化学特性是当前食品和药物管理局（FDA）大多数的标志- 批准的中枢神经系统渗透小分子药物。我们将使用基于结构的设计来促进化学反应策略，我们将通过一系列生化和细胞分析对化合物进行分类，如我们的研究运营计划（ROP）。随着改进的先导化合物的出现，我们将获得酶的选择性很少，通过全面的激酶组（反应生物学）和受体分析（Riserca）。对于关键线索，我们还将通过测量 ASK1，通过生化和细胞研究评估目标参与寿命下游底物磷酸化。集成到 ROP 中，我们还将通过系列药物代谢研究，包括体外微粒体稳定性评估、CYP450 抑制、溶解度、蛋白质结合和 P-糖蛋白流出，并将领先标准集成到化合物的 ROP 中进展。在目标 2 中，将在小鼠中评估外周和中枢神经系统暴露的高级线索急性剂量范围发现和 28 天毒性研究中的不良反应。我们将测量总毒性效应兽医扫描分析以及主要器官的 TUNEL 染色和组织病理学。最后，我们将使用抑制 MKK6、JNK 和 p38 磷酸化作为 ASK1 抑制的体内报告基因，以量化最大生物有效剂量并通过动力学研究建立了我们的领先药物的 PK/PD 关系体内探针。我们位于 TSRI 佛罗里达斯克里普斯校区的多 PI 研究团队拥有专业知识和进行这些实验的基础设施，我们认为成功完成我们的研究将鉴定一系列有效的、选择性的和体内活性的分子探针，这将有助于确定其作用 ASK1 在正常生理过程和疾病状态中发挥作用。