IDENTIFICATION OF IN VIVO PAK1 PROTEIN KINASE INHIBITORS

体内 PAK1 蛋白激酶抑制剂的鉴定

• 批准号: 7609717
• 负责人: Alexander Edward Beeser
• 金额: $ 5.66万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2008-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7609717
• 关键词: Binding; Cell Culture System; Complement; Computer Retrieval of Information on Scientific Projects Database; Engineering; Funding; Genetic; Grant; Growth; Human Pathology; In Vitro; Institution; Link; Malignant Neoplasms; Numbers; Phosphotransferases; Protein Kinase; Protein Kinase Inhibitors; Proteins; Recombinants; Research; Research Personnel; Resources; Signal Transduction Pathway; Source; Specificity; System; Testing; Therapeutic; United States National Institutes of Health; clinically relevant; in vivo; inhibitor/antagonist; kinase inhibitor; member; protein kinase inhibitor; small molecule; small molecule libraries

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. As key regulators of signal transduction pathways, protein kinases are misregulated in a variety of human pathologies. Accordingly protein kinases have long been considered excellent targets for inhibition by small molecules. However, the number of small molecules that target clinically relevant kinases is still quite low. This results predominantly from two considerations; 1) most kinase inhibitors are competitive with ATP and 2) the extent of evolutionary conservation of the ATP binding domain within protein kinases. This means that compounds identified as exquisitely potent inhibitors of a particular kinase in vitro may inhibit a broad number of kinases in vivo. Demonstrating in vivo specificity, a prerequisite for any translational therapeutic, remains a significant obstacle. Pak1 represents an attractive target as it is directly implicated in malignancy and its mechanism of activation allows for the isolation of ATP noncompetitive inhibitors. Paks are members of the Ste20 superfamily of kinases, and complement Ste20 deletions in S.cerevisiae. I have engineered a strain of S.cerevisiae in which growth is directly linked to the kinase activity of ectopic Pak1. I propose to use this platform to screen the small molecule libraries at the CCET for compounds that inhibit Pak1-dependent growth in vivo and to use the power of genetics to counter-select against compounds that target unrelated proteins. I propose confirm specificity by testing promising compounds identified from this platform against recombinant Pak1 in vitro and to further transition these compounds cell culture systems to evaluate their ability to inhibit in higher systems.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 作为信号转导途径的关键调节因子，蛋白激酶在多种人类病理中受到错误调节。 因此，蛋白激酶长期以来被认为是小分子抑制的极好靶标。然而，针对临床相关激酶的小分子数量仍然相当低。 这主要是出于两个考虑； 1) 大多数激酶抑制剂与 ATP 竞争，2) 蛋白激酶内 ATP 结合域的进化保守程度。 这意味着在体外被鉴定为特定激酶的极其有​​效的抑制剂的化合物可以在体内抑制多种激酶。 展示体内特异性（任何转化治疗的先决条件）仍然是一个重大障碍。 Pak1 是一个有吸引力的靶点，因为它直接涉及恶性肿瘤，并且其激活机制允许分离 ATP 非竞争性抑制剂。 Paks 是 Ste20 激酶超家族的成员，可补充酿酒酵母中 Ste20 的缺失。我设计了一种酿酒酵母菌株，其生长与异位 Pak1 的激酶活性直接相关。我建议使用这个平台来筛选 CCET 的小分子库，寻找体内抑制 Pak1 依赖性生长的化合物，并利用遗传学的力量来反选择针对不相关蛋白质的化合物。 我建议通过在体外针对重组 Pak1 测试从该平台鉴定出的有前途的化合物来确认特异性，并进一步转变这些化合物的细胞培养系统以评估它们在更高系统中的抑制能力。