Beta-Secretase Assays to Discover Drugs for Alzheimer's

描述（由申请人提供）：阿尔茨海默病（AD）是一种使人衰弱的疾病， 导致认知功能逐渐丧失的精神紊乱。 然而，对于这种疾病没有有效的治疗方法。神经毒素AB 肽在AD中起着核心作用，涉及增加的生产和 AB在脑淀粉样斑块中的积累。的蛋白水解加工 淀粉样前体蛋白（APP）是产生AB所必需的，这需要 β-分泌酶裂解APP产生AB。有力的证据表明， B-分泌酶的抑制剂应该阻断AB的产生， 治疗AD。8-分泌酶抑制剂药物筛选的关键是 用于真实β-分泌酶活性的快速高通量格式化测定 从天然的体内组织来源获得，具有互补的基于细胞的 从相同的体内组织来源测定。在第一阶段赠款期间， 在分离的嗜铬细胞中鉴定出内源性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 囊泡含有产生AB所需的所有已知组分 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-分泌酶抑制剂。第一阶段的资助开发了嗜铬囊泡 用于筛选B-分泌酶和AB抑制剂的两种测定法 生产：（I）敏感和快速的高通量体外测定， 真实的B-分泌酶活性，和（2）基于嗜铬细胞的测定，以评估 抑制剂对细胞AB产生的影响。第二阶段提案将 使用这些分析来筛选结构多样和集中的组合 选择性抑制B-分泌酶活性和AB 生产将在初始动物中检测这些试验的药物“命中” 安全性和药代动力学特性的毒理学研究，以及 在AD转基因小鼠模型中减少AB的初步研究。这 因此，第二阶段项目将提供B-分泌酶抑制剂的先导化合物 将与制药合作伙伴一起开发新药， 老年痴呆症 拟定商业应用： 该二期项目的临床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估计市场价值为500亿美元，而且还在上升。用独特的高通量8-分泌酶分析和基于细胞的染色质分析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II期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.