Identification of novel system xc- inhibitors

新型系统 xc-抑制剂的鉴定

基本信息

批准号：
8359138
负责人：
Barbara Stauch Slusher
金额：
$ 20.25万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-30 至 2014-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8359138
关键词：
Alzheimer&apos s Disease Amino Acid Transporter Animal Model Antioxidants Astrocytes Biological Assay Biological Factors Brain Neoplasms Cell membrane Cells Chemicals Clinical Research Couples Cystine Dependency Detection Development Dimethyl Sulfoxide Disease Dose Epilepsy Extracellular Space Future Genetic Glioma Glutamate Transporter Glutamates Glutathione Goals Growth HIV In Vitro Inhibitory Concentration 50 Laboratories Lead Libraries Malignant Neoplasms Malignant neoplasm of brain Measurement Metabolism Microglia Molecular Multiple Sclerosis Nerve Degeneration Neurocognitive Neurodegenerative Disorders Neurons Oxidation-Reduction Parkinson Disease Pathogenesis Pharmaceutical Preparations Play Property Proteins Reaction Reactive Oxygen Species Regulation Reporting Role Running Screening procedure Seizures Signal Transduction Staging Structure-Activity Relationship Sulfasalazine System Testing Therapeutic Time Tissues Toxic effect Translating Tumor Expansion Up-Regulation Validation Variant analog antiporter cancer cell cancer therapy cell growth cell type drug discovery experience extracellular ginsenoside M1 high throughput screening in vivo inhibitor/antagonist macrophage miniaturize neuron loss neuroprotection novel pharmacophore prototype response small molecule small molecule libraries therapeutic target tumor growth uptake

项目摘要

DESCRIPTION (provided by applicant): The overall aim of this proposal is to develop a 96-well plate screening assay that will allow screening for small molecule inhibitors of the cystine-glutamate antiporter, also known as system Xc-. System Xc- is a Na+-independent amino acid transporter that couples the export of intracellular L-glutamate with the import of extracellular L cystine. Cystine transport is critical for glutathione synthesis, an intracellular antioxidant whic protects cells from reactive oxygen species. Decreased glutathione levels lead to cellular growth arrest which has made system Xc- a target in cancer therapy, including brain cancer. In brain cancers, the release of glutamate via the antiporter raises additional concerns, as high levels of extracellular glutamate leads to peritumoral neuronal cell death aiding in tumor expansion by clearing surrounding tissue space. In addition to brain cancer, excessive glutamate release via system Xc- has also been shown to play a significant role in several neurodegenerative diseases wherein excess glutamate is presumed pathogenic, including Parkinson's disease, Alzheimer's disease, Multiple Sclerosis and epilepsy, which makes system Xc- a broad therapeutic target for neurodegenerative disorders. There are no selective and potent small molecule system Xc- inhibitors available to the public, and to our knowledge no high throughput screening for this target has been performed. As such, we propose to develop an HTS-ready 96-well assay to screen for novel chemical entities that inhibit system Xc- activity. We will validate this assay by screening four chemical libraries composed of clinically approved drugs as well as non-clinical compounds. The hits will be confirmed in a secondary screening assay and active novel compounds will be moved into structure activity relationship (SAR) studies guided by Xc- potency and in vitro drug- ability assays of metabolism, toxicity and target selectivity. As a future goal, lead compounds arising from these studies will be tested in animal models of glioma and multiple sclerosis and the validated 96-well assay will be further miniaturized to a 384-well format for high throughput screening of the NIH's >300 K compound library. PUBLIC HEALTH RELEVANCE: System Xc- is up-regulated in a variety of cancers and neurodegenerative diseases, and its inhibition with prototype small molecules or genetic deletion inhibits tumor growth in vivo and provides protection in animal models of neurodegeneration. There are no selective and potent small molecule system Xc- inhibitors modulators available to the public, and to our knowledge no high throughput screening (HTS) for this target has been performed. As such, we propose to develop an HTS-ready 96-well assay, screen multiple chemical libraries, and advance hits into a structure activity relationship (SAR) studies guided by Xc- potency and in vitro drug-ability assays of metabolism, toxicity and target selectivity.

描述（由申请人提供）：该提案的总体目的是开发96孔板筛选测定法，该测定法可以筛选胱氨酸 - 谷氨酸抗磷酸盐抗植物的小分子抑制剂，也称为System XC-。系统XC-是Na+独立的氨基酸转运蛋白，将细胞内L-谷氨酸的导出与细胞外L的进口胱氨酸。胱氨酸转运对于谷胱甘肽合成至关重要，细胞内抗氧化剂WHIC可保护细胞免受活性氧的影响。谷胱甘肽水平降低导致细胞生长停滞，这使系统XC-癌症治疗（包括脑癌）成为目标。在大脑癌中，抗精神释放谷氨酸引起了其他关注，因为高水平的细胞外谷氨酸盐会通过清除周围的组织空间清除肿瘤扩张中的肿瘤周神经元细胞死亡。 In addition to brain cancer, excessive glutamate release via system Xc- has also been shown to play a significant role in several neurodegenerative diseases wherein excess glutamate is presumed pathogenic, including Parkinson's disease, Alzheimer's disease, Multiple Sclerosis and epilepsy, which makes system Xc- a broad therapeutic target for neurodegenerative disorders.公众没有选择性且有效的小分子系统XC抑制剂，据我们所知，没有对该目标进行高吞吐量筛选。因此，我们建议开发可预见的HTS准备的96孔测定法，以筛选抑制系统XC活性的新型化学实体。我们将通过筛选四个由临床认可的药物以及非临床化合物组成的化学文库来验证该测定法。次要筛选分析将确认命中，并将活跃的新颖化合物转移到以XC-效力和体外药物能力分析为指导的结构活动关系（SAR）研究中，该研究的代谢，毒性和靶标选择性。作为未来的目标，这些研究产生的铅化合物将在神经胶质瘤和多发性硬化症的动物模型中进行测试，并且经过验证的96孔测定法将进一步微型化，以384孔格式进行384杆> 300 K的高吞吐量筛选。公共卫生相关性：系统XC-在多种癌症和神经退行性疾病中被上调，并且其对原型小分子或遗传缺失的抑制作用抑制了体内肿瘤的生长，并在神经变性的动物模型中提供了保护。没有选择性且有效的小分子系统XC抑制剂调节剂可供公众使用，据我们所知，该目标尚未进行高吞吐量筛选（HTS）。因此，我们建议开发可提供HTS就绪的96孔测定法，筛选多个化学文库，并将其提高到以XC效力和具有代谢性，毒性和靶向选择性的体外药物能力测定为指导的结构活动关系（SAR）研究中。