Beta-Secretase Assays to Discover Drugs for Alzheimer's

描述(申请人提供)：阿尔茨海默病(AD)是一种使人虚弱的疾病 导致认知功能逐渐丧失的精神障碍。 然而，目前还没有针对这种疾病的有效治疗方法。神经毒素AB 多肽在阿尔茨海默病中起核心作用，包括增加产量和 AB在脑淀粉样斑块中的积聚。大豆蛋白水解液的加工工艺 淀粉样前体蛋白(APP)需要生成AB，这需要 B-分泌APP的裂解产生AB。强有力的证据表明 B-分泌酶抑制剂应阻断AB的产生并成为一种有效的 治疗阿尔茨海默病。8-分泌酶抑制剂药物筛选的关键是 3-分泌酶活性的快速高通量格式化检测 从天然的体内组织来源获得，以补充细胞为基础 从相同的体内组织来源进行化验。在第一阶段授权期内， 分离的嗜铬细胞具有内源性B-分泌酶分泌酶活性 神经元嗜铬细胞的小泡(分泌小泡)。这些孤立的 囊泡含有生产AB所需的所有已知成分 由APP、AB(1-40)、B-分泌酶活性、BACE-1和早老素组成； 因此，这些囊泡为筛选提供了理想的模型系统 B-分泌酶的抑制剂。第一期授权者发育出嗜铬小泡 两种方法筛选B-分泌酶和AB抑制剂的体系 生产：(I)灵敏、快速、高通量的体外检测方法 真实的B-分泌酶活性，以及(2)以嗜铬细胞为基础的检测 抑制剂对细胞AB产生的影响。这项第二阶段的提案将 使用这些分析来筛选结构多样化和重点突出的组合 选择性抑制B-分泌酶活性和AB的化合物文库 制作。来自这些测试的药物“命中”将在最初的动物身上进行测试 安全性和药代动力学特性的毒理学研究，以及 在AD转基因小鼠模型中降低AB的初步研究。这 因此，第二阶段项目将为B-分泌酶抑制剂提供先导化合物 将与制药合作伙伴共同开发用于治疗的新药 阿尔茨海默氏症。 建议的商业应用： 这一第二阶段项目的商业应用将是开发治疗阿尔茨海默病的新药。一种有效的阿尔茨海默氏症药物的市场价值估计为500亿美元，而且还在上升。用独特的高通量8-分泌酶试验和基于嗜铬细胞的试验筛选组合文库将为阿尔茨海默病提供新的先导化合物。ActiveSite Biotech能够独家销售、许可或合作伙伴第二阶段研究产生的主要药物抑制剂。

DESCRIPTION (provided by applicant): Alzheimer's disease (AD) is a debilitating mental disorder that results in a progressive loss of cognitive functions. However, there is no effective treatment for this disease. The neurotoxin AB peptide plays a central role in AD, involving increased production and accumulation of AB in brain amyloid plaques. Proteolytic processing of the amyloid precursor protein (APP) is required to generate AB, which requires B-secretase cleavage of APP to produce AB. Strong evidence indicates that inhibitors of B-secretase should block AB production and be an effective treatment for AD. The key to drug screening for inhibitors of 8-secretase is a rapid high throughput formatted assay for authentic 3-secretase activity obtained from a natural, in vivo tissue source, with a complementary cell-based assay from the same in vivo tissue source. In the phase I grant period, endogenous B-secretase secretase activity was identified in isolated chromaffin vesicles (secretory vesicles) of neuronal chromaffin cells. These isolated vesicles contain all the known components required for production of AB consisting of APP, AB(1-40), B-secretase activity, BACE 1, and presenilin; these vesicles, therefore, provide an ideal model system for screening inhibitors of B-secretase. The Phase I grant developed the chromaffin vesicle system into two assays for screening inhibitors of B-secretase and AB production: (I) sensitive and rapid high throughput in vitro assays of authentic B-secretase activity, and (2) chromaffin cell-based assays to assess effects of inhibitors on cellular AB production. This Phase II proposal will use these assays to screen structurally diverse and focused combinatorial libraries for compounds that selectively inhibit B-secretase activity and AB production. Drug "hits" from these assays will be tested in initial animal toxicology studies for safety and pharmacokinetic properties, and in preliminary studies for reduction of AB in a transgenic mouse model of AD. This phase II project will, thus, provide lead compounds for B-secretase inhibitors that will be developed with pharmaceutical partners for new drugs to treat Alzheimer's disease. PROPOSED COMMERCIAL APPLICATION: The commerical application of this Phase II project will be development of novel drugs for Alzheimer's disease. The estimated market value for an effective Alzheimer's drug is $50 billion dollars and rising. Screening of combinatorial libraries with the unique high throughput 8-secretase assay, and chromaffm cell-based assay will provide new lead compounds for Alzheimer's disease. ActiveSite Biotech is in a position to exclusively sell, license, or partner lead drug inhibitors resulting from the Phase II study.