PKC Modulators for the Treatment of Alzheimer's disease

描述（由申请人提供）：越来越多的人认为阿尔茨海默病（AD）病理生理学中的关键事件是淀粉样β肽（Abeta）的过度产生。如果这个公式是正确的，那么有三个主要的策略来中断病理级联反应：抑制β-分泌酶，抑制γ-分泌酶，或增加α-分泌酶。直觉上，如果Abeta是罪魁祸首，减少肽的产生是最有希望的方法。β-和γ-分泌酶都是产生完整的Abeta所必需的，而α-分泌酶在Abeta结构域内切割，阻止其完整释放。制药业在β-和γ-分泌酶抑制剂的开发上投入了大量资金，尽管还没有一种在人体试验中取得成功。我们提出了一个战略的基础上的第三种方法，增强α-分泌酶活性。我们增加α-分泌酶活性的方法涉及调节蛋白激酶C（PKC），这在许多实验室中已被确定与α-分泌酶的调节有关。在这方面，我们最近已经表明，结构新颖的PKC激活剂，8-（1-癸炔基）-苯并内酰胺，是能够扭转K+通道缺陷，并提高生产的α-分泌酶产品sAPP α在AD细胞。更重要的是，在体内测试时，这种苯并内酰胺显著增加了APP[V7171]转基因小鼠大脑中sAPP α的量，并减少了A β 40。鉴于8-（1-癸炔基）-苯并内酰胺获得的有希望的动物数据和我们在苯并内酰胺（BL）SAR方面的长期经验，我们建议进行一项高度集中的研究，旨在确定可作为治疗AD的临床候选药物的最佳苯并内酰胺类似物。为了实现这一目标，我们的目标如下： 1.使用分子建模工具进行化合物设计和新型BL的化学合成，目的是优化对α-分泌酶活性的影响，同时减少任何可能的肿瘤促进活性;这项工作的大部分将集中在改变侧链附件的性质; 2.进行体外研究以确定新类似物的生物活性。筛选所有化合物作为PKC激活剂的活性。测试选定的有效PKC激活剂（EC 50至少50 nM）对sAPPa和Abeta 1-40和1-42水平的影响。有效增强sAPP α产生以产生肿瘤促进作用的测试化合物; 3.对于来自目标2的最佳化合物，在三重转基因小鼠中进行研究，以确定对体内Abeta和sAPP α水平和斑块形成的影响。

DESCRIPTION (provided by applicant): There is a growing consensus that the pivotal event in the pathophysiology of Alzheimer's disease (AD) is overproduction of amyloid beta peptide (Abeta). If this formulation is correct, then there are three primary strategies for interrupting the pathologic cascade: inhibiting beta-secretase, inhibiting gamma-secretase, or augmenting alpha-secretase. Intuitively, if Abeta is the culprit, reducing the generation of the peptide is the most promising approach. Both beta- and gamma-secretase are necessary for generation of intact Abeta, while alpha-secretase cleaves within the Abeta domain, preventing its release intact. The pharmaceutical industry has invested heavily in the development of beta- and gamma-secretase inhibitors, though none has yet succeeded in human trials. We propose a strategy based on the third approach, augmenting alpha-secretase activity. Our approach to augmenting alpha-secretase activity involves modulating protein kinase C (PKC), which has conclusively been linked to regulation of alpha-secretase in a number of laboratories. In this regard, we have shown recently that the structurally novel PKC activator, 8-(1-decynyl)-benzolactam, is able to reverse K+ channels defects and to enhance the production of the alpha-secretase product sAPPalpha in AD cells. More importantly, when tested in vivo, this benzolactam significantly increased the amount of sAPPalpha and reduced Abeta40 in the brains of APP[V7171] transgenic mice. Given the promising animal data achieved with the 8-(1-decynyl)-benzolactam and our long experience in the SAR of the benzolactams (BL), we propose to conduct a highly focused study aimed at defining the best benzolactam analog that can serve as a clinical candidate for the treatment of AD. To achieve this goal, our aims become the following: 1. Conduct both compound design using molecular modeling tools and the chemical synthesis of novel BLs with the aim to optimize effects on alpha-secretase activity while diminishing any possible tumor promoting activity; Much of this work will focus on altering the nature of the side chain appendages; 2. Carry out in vitro studies to define the biological activity of new analogs. Screen all compounds for activity as PKC activators. Test selected potent PKC activators (EC50 at least 50 nM) on levels of sAPPalpha and Abeta 1-40 and 1-42. Test compounds effective in enhancing sAPPalpha production for tumor promoting effects; 3. For the best compounds from Aim 2, perform studies in triple transgenic mice to ascertain effects on Abeta and sAPPalpha levels and plaque formation in vivo.