SYNTHESIS AND SPECTROSCOPY OF PHOSLIPID ANALOGUES

磷脂类似物的合成和光谱学

• 批准号: 6204256
• 负责人: JOSEPH HAJDU
• 金额: $ 14.76万
• 依托单位: CALIFORNIA STATE UNIVERSITY NORTHRIDGE
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-07-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6204256
• 关键词: analog; analytical method; cell membrane; chemical addition; chemical structure function; chemical synthesis; electron spin resonance spectroscopy; enzyme activity; enzyme structure; enzyme substrate; fluorescence spectrometry; lipid structure; membrane model; nuclear magnetic resonance spectroscopy; oxidizing agents; phosphodiesterases; phospholipase A2; phospholipase inhibitor; phospholipids; stereochemistry

Development of new synthetic methods for the preparation of structurally well-defined phospholipid compounds has become a timely and important problem since it was discovered that in addition to their role as a major component in all biological membranes phospholipids are also involved in a wide range of physiological and regulatory processes. Phospholipids are essential for the functioning of a number of membrane-bound enzymes such as protein kinase C, and play an important role in signal transduction as substrates for the production of second messengers (inositol-1,4,5-trisphophate, diacylglycerol) and in the release of arachidonic acid. Significantly, some of the most potent phospholipid compounds, which are active below the micromolar and nanomolar levels occur in low concentrations in the cell, such that preparation of synthetic derivatives represents a clear prerequisite to elucidation of their biochemical mechanism of action. The main objective of the proposed project is to develop new synthetic methods for the preparation of phospholipid derivatives, including compounds with spectroscopically active reporter groups and their characterization as structural probes in membrane models (micelles and vesicles). The approach will focus on developing the chemistry of the five-membered 1,3,2-dioxaphospholane ring and its substituted derivatives to elaborate phosphodiestertargets and structurally related analogues. The synthetic application will focus on preparation of 1) diacylglycerophospholipid spectroscopic probes with paramagnetic and fluorescence energy transfer donor-acceptor reporter groups, 2) transition-state analogue inhibitors of phospholipase A2 enzymes, and 3) boranophopholipids with a BH3 group replacing one of the nonbriding phosphate-oxygens in the phosphodiester function. Electron spin resonance studies using a newly developed "indicator broadener method" will be conducted in the principal co-investigator's to determine the structural organization and dynamics of fatty acyl side chains of single- and double-labeled phopholipids. Collaborative arrangements have been made to evaluate the synthetic compounds as phospholipase A2 substrates and inhibitors. In addition, their thermotropic behavior and role in promoting vs. Inhibiting cholesterol exchange between vesicles will also be determined.

合成新方法的开发 结构明确的磷脂化合物已成为一个及时的 这是一个重要的问题，因为人们发现，除了他们的 磷脂作为所有生物膜的主要成分， 还参与了广泛的生理和调节过程。 磷脂对许多细胞的功能至关重要， 膜结合酶如蛋白激酶C，并发挥作用， 在信号转导中的重要作用，作为生产 第二信使（肌醇-1，4，5-三磷酸，甘油二酯）和 花生四烯酸的释放。值得注意的是，一些最有效的 磷脂化合物，其在低于微摩尔浓度时具有活性， 纳摩尔水平在细胞中以低浓度出现，使得 合成衍生物的制备是一个明确的先决条件， 阐明其生化作用机制。 主要目标 该项目的目的是开发新的合成方法， 磷脂衍生物的制备，包括具有 光谱活性报道基团及其表征 膜模型中的结构探针（胶束和囊泡）。的 方法将侧重于开发五元化合物的化学性质， 1，3，2-二氧磷杂环戊烷环及其取代衍生物进行阐述 磷酸二酯靶标和结构相关的类似物。合成 应用将集中在1）二酰基甘油磷脂的制备 具有顺磁和荧光能量的光谱探针 转移供体-受体报告基团，2）过渡态类似物 磷脂酶A2酶的抑制剂，和3）硼磷脂， BH3基团取代了其中一个非桥连的磷酸氧， 磷酸二酯功能。 电子自旋共振研究，使用新的 制定的"指标扩大方法"将在 主要合作研究者确定结构组织， 单标记和双标记的脂肪酰基侧链的动力学 磷脂 已作出合作安排， 合成的化合物作为磷脂酶A2的底物和抑制剂。 此外，还研究了它们的热致性行为和促进作用。 还将测定抑制囊泡之间的胆固醇交换。