Synthetic Study on Neurotrophins and Its Application for Development of Diagnostic Immunoassay Method for Senile Dementia

为建立老年性痴呆的免疫诊断方法，进行了以下研究。否定血浆神经营养因子水平与阿尔茨海默病之间的显著相关性的报道迫使我们改变了最初的计划，然而，本文所进行的基础研究应有助于未来神经营养因子对老年性痴呆的预防和/或治疗用途的研究。提出了一种基于双功能巯基引入试剂的合成肽化学选择性纯化新方法。此外，还建立了利用可逆生物素化试剂的特异性亲和纯化方法，用于固相技术对合成肽的最终纯化.结合新发展的合成肽的化学选择性纯化方法和固相片段缩合法，对神经原蛋白进行了合成研究。然而，含Cys的肽片段的低纯化产率和低的纯化产率是不可行的。 ...更多信息 在固相片段缩合中的扭曲产率沿着链延长迫使我们放弃神经营养素的全合成。为了促进含有三个二硫键的神经营养因子的最终二硫键形成，开发了新的亚砜介导的二硫键形成反应，并成功地应用于几种生物活性肽（包括抗HIV肽和刺激NGF合成的记忆增强肽）的明确的区域选择性侧硫键形成.为了研究神经营养因子Trk受体的胞内信号转导途径和与阿尔茨海默病相关的Tau蛋白异常磷酸化，建立了一种高效的固相合成磷酸肽的方法。此外，研究了抗磷酸酶的磷酸氨基酸模拟物和非水解二肽模拟物（E）-烯烃二肽异构体的立体选择性合成方法。这些非水解磷酸肽模拟物将作为有用的工具，生物化学研究蛋白磷酸化与阿尔茨海默病。少

The following investigations were caried out for development of diagnostic immunoassay method for senile dementia. The report which deny the significant correlation between the plasma levels of neurotrophins and Alzheimer's disease forced us to change the initial plan, however, basic study conducted here should contribute to the future research on daignostic and/or therapeutic use of neurotrophins for senile dementia.1. A new chemoselective purication method of synthetic peptides based on the bifunctional SH-introducing reagent was developed. In addition, specific affinity purification method using a reversible biotinylating reagent was developed for the final purification of synthetic peptides by solid phase technique.2. Synthetic study on neuroprophins was carried out in combination with newly-developed chemoseletcive purification method of synthetic peptides and fragment condensation method on solid support. However, low purification yields of Cys-containing peptide fragments and lo … More wering yields in solid phase fragment condensations along with chain elongation compelled us to abandon the total synthesis of neurotrophins.3. In order to facilitate the final disulfide bond formation of neurotrophins containing three disulfide bonds, new sulfoxide-mediated disulfide bond- forming reactions were developed and successfully applied for unambiguous regioselectie sidulfide-bond formation of several biologically active peptides including anti-HIV peptides and memory enhancing peptides which stimulate NGF synthesis.4. In order to investigate the intracellular signal transductin pathway of neurotrophin Trk-receptors and the abnormal phosphorylations of Tau-protein associated with Alzheimer's Disease, an efficient synthetic method of phosphopeptides by solid phase technique was developed. Furthermore, stereoselective synthetic methods of phosphatase-resisted phosphoaminoacids mimetics and (E) -alkene-dipeptide isoseres as nonhydrolyzable dipeptide mimetics were investigated. These nonhydrolyzable phosphopeptide mimetics will serve as useful tools for biochemical study on protein phosphorylation associated with Alzheimer's disease. Less