Gamma Secretase Assays to Discover Drugs for Alzheimer's

伽马分泌酶检测发现治疗阿尔茨海默病的药物

• 批准号: 7101003
• 负责人: GREGORY R HOOK
• 金额: $ 80.84万
• 依托单位: AMERICAN LIFE SCIENCE PHARMACEUTICALS
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-30 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7101003
• 关键词: Alzheimer' s disease; amyloid proteins; animal tissue; aspartic endopeptidases; bioassay; blood brain barrier; chemical structure function; chromaffin cells; drug discovery /isolation; drug screening /evaluation; enzyme activity; fluorescent dye /probe; high throughput technology; neuritic plaques; neurons; neuropharmacology; nicotine; potassium chloride; protease inhibitor; technology /technique development; tissue /cell culture; vesicle /vacuole

DESCRIPTION (provided by applicant): Alzheimer's disease (AD) is a long-term, debilitating disorder that causes loss of memory and cognitive functions. There is an urgent need for effective therapeutic agents to ameliorate the symptoms of AD. Important molecular drug targets in AD are the gamma-secretases that produce the neurotoxic Abeta peptides in AD. The regulated secretory pathway of neurons represents the major source of secreted Abeta peptides that accumulate in extracellular amyloid plaques in AD. However, previous screens for inhibitors of gamma-secretase have only analyzed the minor constitutive secretory pathway for Abeta peptide production. Different proteases are present in the regulated secretory pathway compared to the constitutive secretory pathway. Clearly, gamma-secretase in the regulated secretory pathway must be targeted for drug inhibition to provide the greatest reduction of Abeta. Therefore, the Phase I project developed high-throughput assays for gamma-secretases in regulated secretory vesicles that produce Abeta. A candidate inhibitor of gamma-secretases was identified, which will provide the basis for design and synthesis of 'focused' and 'optimized' libraries for this Phase II project. Phase I results also indicate feasibility to find agents that selectively inhibit gamma-secretases-42 compared to gamma-secretases-40. The Phase I project also developed neuronal chromaffin cell and brain cortical neuron assays for Abeta in the regulated secretory pathway. The goal of this phase II project will be to utilize the regulated secretory vesicle as the major site and target of Abeta peptide production in neuronal cells, for identifying inhibitor molecules that selectively inhibit gamma-secretases-42 compared to gamma-secretases-40. Such inhibitors will be considered as lead compounds. In specific aim 1, efforts for 'focused' library compounds will be screened in the high throughput in vitro assays to identify inhibitors of gamma-secretases-40 and gamma-secretases-42 in chromaffin vesicles; inhibitors will then be tested in chromaffin cells for reduction of Abeta. In the second aim, further 'optimized' libraries will be designed and synthesized based on structural features of inhibitors from the 'focused' library, screened in the high throughput assays, and tested for reduction of Abeta in the regulated secretory pathway of neuronal chromaffin cells. In the third aim, lead compounds will undergo evaluation in brain neuronal cells to identify compounds that reduce production of Abeta peptides in the regulated secretory pathway. Results will likely identify novel lead compounds for future pre-clinical animal studies for development of effective drugs for AD.

描述（由申请人提供）：阿尔茨海默病（AD）是一种导致记忆和认知功能丧失的长期衰弱性疾病。目前迫切需要有效的治疗药物来改善阿尔茨海默病的症状。阿尔茨海默病的重要分子药物靶点是在阿尔茨海默病中产生神经毒性肽的γ -分泌酶。神经元受调节的分泌途径是阿尔茨海默病细胞外淀粉样斑块中积累的β肽分泌的主要来源。