Validating Inhibitors of the Immunomodulatory Sphingosine 1-Phosphate Transporter SPNS2

验证免疫调节鞘氨醇 1-磷酸转运蛋白 SPNS2 的抑制剂

基本信息

批准号：
10348768
负责人：
Susan Ruth Schwab
金额：
$ 27.2万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-03-01 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10348768
关键词：
Autoimmune Autoimmune Diseases Autoimmune Responses Biological Biological Assay Biological Process Biology Blood Blood Vessels Bradycardia Cardiac Cardiac Myocytes Cardiovascular Physiology Cardiovascular system Cells Chemicals Clinical Colitis Complex Data Dose Drug Targeting Endothelial Cells Erythrocytes Exhibits FDA approved Fire - disasters Future Generations Goals Grant Human Immune system Immunity In Vitro Inflammation Inorganic Phosphate Transporter Laboratories Lead Libraries Lipids Lymph Lymphatic Endothelial Cells Lymphocyte Mass Spectrum Analysis Multiple Sclerosis Mus Neuraxis Oral Pathogenicity Pathway interactions Patients Pharmaceutical Chemistry Pharmaceutical Preparations Phase II Clinical Trials Play Process Property Psoriasis Recombinants Research Contracts Rivers Role Running SPHK1 enzyme Scheme Series Services Signal Transduction Site Species Specificity Specificity Sphingosine-1-Phosphate Receptor Synthesis Chemistry System T-Lymphocyte Testing Therapeutic Therapeutic Agents Tissues Travel Validation Work analog assay development base cell type clinically relevant counterscreen drug development drug discovery fighting high throughput screening immunoregulation improved in vivo inhibitor lymph nodes lymphoid organ macular edema mouse model multiple sclerosis treatment new therapeutic target pathogen prevent protective effect response scaffold screening secondary lymphoid organ side effect small molecule sphingosine 1-phosphate sphingosine kinase targeted treatment tool

项目摘要

Project Summary The signaling lipid sphingosine 1-phosphate (S1P) plays critical roles in the immune system. Notably, S1P regulates lymphocyte exit from lymph nodes, where lymphocytes are initially activated to fight pathogens, out into lymph, which carries the cells to blood and enables them to travel to infected tissues. FTY720, a drug that targets four S1P receptors including S1PR1, became the first FDA-approved oral therapeutic for multiple sclerosis. By blocking pathogenic T cells from exiting lymphoid organs, FTY720 prevents them from accessing the central nervous system. Second-generation drugs that target S1PR1 have also shown promise in Phase II clinical trials for psoriasis and colitis. Unfortunately, these drugs also target S1PR1 in endothelial cells and cardiomyocytes, resulting in serious side effects including macular edema and bradycardia. We found that the major facilitator superfamily transporter SPNS2 supplies S1P into lymph but not blood, making it a promising new drug target that would enable spatially specific modulation of S1P signaling. Furthermore, our data indicate that Spns2 is required for the accumulation of activated T cells at sites of inflammation and deleting Spns2 has a protective effect in experimental autoimmune encephalomyelits, a mouse model of multiple sclerosis. Based on these observations, we set out to develop a high throughput screen to identify small molecule modulators of Spns2 activity. We developed and optimized a Rapid-Fire high-throughput mass spectrometry-based assay to detect S1P secreted into the media by Spns2-expressing cells. Utilizing this assay, we screened Evotec's 252,454 compound Discovery Library, yielding 5,899 inhibitors at a cut-off of 11.5% (5σ). We subsequently cherry picked 1962 hits for confirmation screens, yielding 923 confirmed hits. Here, we propose to develop assays and validate the hits that emerged from the HTS effort. Together with NYU's Office of Therapeutics Alliances we have identified a team of consultants and contract research organizations with expertise in assay development and screening, hit validation, and synthetic and medicinal chemistry that will work with our laboratory to develop and execute a hit validation scheme yielding high quality probes targeting Spns2. These tool compounds can be used to probe the biology of Spns2 and have the potential to yield first in class drugs for treating autoimmune diseases.

项目概要信号脂质 1-磷酸鞘氨醇 (S1P) 在免疫系统中发挥着关键作用。 S1P 调节淋巴细胞从淋巴结排出，淋巴细胞最初被激活以对抗病原体，进入淋巴液，淋巴液将细胞带到血液中，并使它们能够到达受感染的组织 FTY720（一种药物）。靶向包括 S1PR1 在内的四种 S1P 受体，这是 FDA 批准的第一个针对多种疾病的口服治疗药物 FTY720 通过阻止致病性 T 细胞离开淋巴器官，阻止它们进入淋巴器官。靶向 S1PR1 的第二代药物在 II 期临床试验中也显示出前景。不幸的是，这些药物还针对内皮细胞和结肠炎中的 S1PR1。心肌细胞，导致严重的副作用，包括黄斑水肿和心动过缓。主要促进超家族转运蛋白 SPNS2 将 S1P 输送到淋巴液中，但不输送到血液中，这使其成为一种有前途的药物新的药物靶标能够实现 S1P 信号的空间特异性调节此外，我们的数据。表明 Spns2 是活化 T 细胞在炎症部位积聚和删除所必需的 Spns2 对实验性自身免疫性脑脊髓炎（一种患有多种疾病的小鼠模型）具有保护作用根据这些观察结果，我们着手开发一种高通量筛选来识别微小的硬化症。 Spns2 活性的分子调节剂我们开发并优化了 Rapid-Fire 高通量质量。利用基于光谱分析的方法检测 Spns2 表达细胞分泌到培养基中的 S1P。分析中，我们筛选了 Evotec 的 252,454 种化合物发现库，以 11.5% 的截止值产生了 5,899 种抑制剂 (5σ)。我们随后挑选了 1962 个命中进行确认屏幕，在这里，我们产生了 923 个确认命中。与纽约大学办公室一起开发分析方法并验证 HTS 的建议。我们已经确定了一个由顾问和合同研究组织组成的团队检测开发和筛选、命中验证以及合成和药物化学方面的专业知识与我们的实验室合作开发并执行命中验证方案，产生高质量的探针靶向这些工具化合物可用于探索 Spns2 的生物学，并有可能在治疗自身免疫性疾病的类。