Comput. Struct.-Based Design, Chem. Synth., Biochem Charac. and Molec. Mechanism

计算。

• 批准号: 6934229
• 负责人: SHAOMENG WANG
• 金额: $ 28.62万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6934229
• 关键词: BCL2 gene /protein; antineoplastics; apoptosis; chemical synthesis; computational biology; cooperative study; drug design /synthesis /production; drug discovery /isolation; fluorescence polarization; gene induction /repression; inhibitor /antagonist; neoplastic cell; nuclear magnetic resonance spectroscopy; pharmacokinetics; protein binding; protein protein interaction; protein structure function; small molecule; tissue /cell culture

This NCDDG application proposes the design and development of novel nonpeptidic, small-molecule inhibitors of Bcl-2 and Bcl-xL proteins as a new class of anti-cancer drugs through a contemporary, multidisciplinary and integrated drug discovery approach. The first primary goal of Laboratory Program #1 is the design and synthesis of highly potent and optimized small-molecule inhibitors of Bcl-2/Bcl-xL through a structure-based design approach. The second primary goal is the characterization of their binding affinities to Bcl-2 and Bcl-xL proteins in biochemical binding assays. The third primary goal is the characterization of their activity in cancer cells with high levels of Bcl-2 and/or Bcl-xL protein and their specificity to normal cells. The fourth primary goal is the elucidation of the molecular mechanisms of action by which they induce apoptosis using isogenic model cell systems. We will perform the following specific Aims: (1). Performance of computational structure-based design for lead optimization based upon the experimental structures of small-molecule inhibitors in complex with Bcl-2 and Bcl-xL, as determined in Laboratory Program #2. (2). Chemical synthesis of new compounds designed in Aim 1. (3). Biochemical characterization for their binding affinities to Bcl-2/xL proteins and their specificity to other proteins using our established fluorescence-polarization (FP)-based binding assays. This is followed by a conclusive confirmation by the NMR Heteronuclear Single Quantum Coherence (HSQC) experiments outlined in Laboratory Program #2 (Aim 1) to rule out any potential false positives. (4). Determination of the activity, specificity and molecular mechanisms by which small molecule inhibitors of Bcl-2/Bcl-xL induce apoptosis. Laboratory Program #1 forms the foundation of this NCDDG grant and provides highly potent and optimized small-molecule inhibitors of Bcl-2/Bcl-xL with a clear understanding of their activity, specificity, and molecular mechanism of action to be studied in Laboratory Program #2 and #3.

该 NCDDG 申请提出通过当代、多学科和综合药物发现方法设计和开发 Bcl-2 和 Bcl-xL 蛋白的新型非肽小分子抑制剂作为一类新型抗癌药物。实验室计划#1 的第一个主要目标是通过基于结构的设计方法设计和合成高效且优化的 Bcl-2/Bcl-xL 小分子抑制剂。第二个主要目标是在生化结合测定中表征它们与 Bcl-2 和 Bcl-xL 蛋白的结合亲和力。第三个主要目标是表征 它们在具有高水平 Bcl-2 和/或 Bcl-xL 蛋白的癌细胞中的活性及其对正常细胞的特异性。第四个主要目标是利用等基因模型细胞系统阐明它们诱导细胞凋亡的分子作用机制。我们将实现以下具体目标： (1)．根据实验室计划 #2 中确定的与 Bcl-2 和 Bcl-xL 复合的小分子抑制剂的实验结构，进行基于计算结构的先导优化设计的性能。 （2）。目标 1. (3) 中设计的新化合物的化学合成。使用我们建立的基于荧光偏振 (FP) 的结合测定法对其与 Bcl-2/xL 蛋白的结合亲和力及其对其他蛋白的特异性进行生化表征。随后通过实验室计划 #2（目标 1）中概述的 NMR 异核单量子相干 (HSQC) 实验进行结论性确认，以排除任何潜在的误报。 （4）。确定 Bcl-2/Bcl-xL 小分子抑制剂诱导细胞凋亡的活性、特异性和分子机制。 实验室计划 #1 构成了 NCDDG 拨款的基础，并提供了高效且 优化的 Bcl-2/Bcl-xL 小分子抑制剂，清楚地了解其活性、特异性和分子作用机制，将在实验室计划 #2 和 #3 中进行研究。