PROBING THE S' SUBSITES OF CALPAIN

简介(申请人提供)：心脏病和中风是主要原因 美国的发病率和死亡率。越来越多的证据表明 心脏缺血(心脏病发作)或脑缺血发作 (中风)引发一系列生化事件，导致 细胞内钙离子浓度，进而激活钙蛋白酶。已激活 钙蛋白酶降解结构蛋白，导致细胞死亡。CalPain是 因此被认为是干预的有吸引力的治疗靶点 心脏病发作和中风。 长期的目标是发现新的钙蛋白酶抑制剂作为治疗 心脏病发作和中风。其具体目标是确定 以开发有效和选择性为目标的钙蛋白酶S亚基 酶的抑制剂。缓蚀剂结合到的结构要求 对CalPain的S亚点进行了深入的研究。恰恰相反， S结合缓蚀剂的结构要求？钙调蛋白亚基 只受到了轻微的调查。这是一个很大的差距， 哪些文献必须填满，因为了解了S？子网站 钙蛋白酶的特异性将提供有价值的信息，有助于 该酶选择性抑制剂的设计。含有天然和天然成分的抑制剂 P1位非天然D-氨基酸和L-氨基酸？那P2呢？抑制剂的位置 会被合成来探测S1的特异性吗？和S2呢？的子网站 钙调蛋白。将使用比较分子场分析(CoMFA)来生成 抑制剂的结合部位模型和该模型将被用于设计 并在化学前预测新型抑制剂的钙蛋白酶抑制效力 合成和酶学。有效的抑制剂将接受测试，以确定它们是否能够 进入细胞并抑制细胞内的钙蛋白酶。选定的有效和细胞 渗透性抑制剂将在大鼠离体心缺血模型中进行测试 对心脏有保护作用。

DESCRIPTION (provided by applicant): Heart disease and stroke are major causes of morbidity and mortality in the United States. Mounting evidence suggests that an episode of cardiac ischemia (heart attack) or cerebral ischemia (stroke) initiate a chain of biochemical events that result in increased intracellular Ca2+ concentration, which in turn activates calpain. Activated calpain degrades structural proteins resulting in cell death. Calpain is therefore considered as an attractive therapeutic target for intervention in heart attack and stroke. The long-term goal is to discover novel calpain inhibitors as treatment for heart attack and stroke. The specific aim is to determine the specificities of the S subsites of calpain with the objective of developing potent and selective inhibitors of the enzyme. The structural requirements for inhibitor binding to the S subsites of calpain has been well investigated. On the contrary, the structural requirements for inhibitor binding to the S? subsites of calpain have only been marginally investigated. This is a significant gap in the literature that must be filled because knowledge about the S? subsite specificities of calpain will provide valuable information that can aid in the design of selective inhibitors of the enzyme. Inhibitors with natural and unnatural D- and L-amino acids at the P1? and P2? positions of the inhibitors will be synthesized to probe the specificities of the S1? and S2? subsites of calpain. Comparative Molecular Field Analysis (CoMFA) will be used to generate a binding site model for the inhibitors and the model will be used to design and predict the calpain inhibitory potency of novel inhibitors before chemical synthesis and enzymology. Potent inhibitors will be tested for their ability to enter cell and inhibit intracellular calpain. Selected potent and cell permeable inhibitors will be tested in the rat isolated heart ischemia model for cardioprotective effect.