Identification of novel system xc- inhibitors

新型系统 xc-抑制剂的鉴定

• 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
• 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' 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

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）：本提案的总体目标是开发一种96孔板筛选试验，该试验将允许筛选胱氨酸-谷氨酸反转运蛋白的小分子抑制剂，也称为系统Xc-。系统Xc-是一种不依赖于Na+的氨基酸转运体，它将细胞内L-谷氨酸的输出与细胞外L的输入偶联