Kinase selective small molecule conjugates as antibody surrogates

描述（由申请人提供）：蛋白激酶广泛涉及人类疾病，特别是癌症，并在人类生物学中发挥基础作用。为了理解所谓的“磷酸化蛋白质组”，人们需要新的工具来确定在给定的信号转导网络中哪些特定的激酶(s)是活跃的（通常是磷酸化的）。目前几乎所有用于鉴定药物诱导表达水平和特定激酶磷酸化状态的蛋白质阵列技术都使用基于抗体的捕获方法。然而，抗体捕获剂价格昂贵，通常不可扩展，最近的报告强烈表明，只有不到30%的可用抗体可以用于微阵列技术。为了提供一种可行的抗体替代品，本提案寻求系统地开发一类新的二价捕获剂（bca），用于连接高亲和力小分子和噬菌体显示肽的激酶。bca具有特定的、可扩展的和经济的生产潜力。这些显著特征使我们的bca作为一种新型试剂具有吸引力，可用于阵列技术，用于理解激酶磷酸化的调控，激酶磷酸化在人类疾病，特别是癌症中受到干扰的许多信号网络中起着核心作用。公共卫生相关性：蛋白激酶是信号转导级联的核心，当信号转导级联受到干扰时，会导致癌症等人类疾病。因此，在微阵列应用中，识别用于捕获激酶的新型二价试剂对理解人类疾病的病理生理学具有直接的效用，并有助于开发新的诊断和治疗方法。

DESCRIPTION (provided by applicant): Protein kinases are widely implicated in human disease, especially in cancer, and play a fundamental role in human biology. In order to understand the so-called "phospho-proteome" one needs new tools to address which particular kinase(s) are active (usually phosphorylated) in a given signal transduction network. Almost all current protein-array technologies for identifying drug-induced expression levels and phosphorylation states of specific kinases utilize antibody based capture methods. However antibody capture agents are expensive, often not scalable, and recent reports strongly suggest that fewer than 30% of available antibodies can be utilized in microarray technologies. In order to provide a viable alternative to antibodies, this proposal seeks to systematically develop a new class of bivalent capture agents (BCAs) for kinases that link high-affinity small molecules to phage-displayed peptides. BCAs have the potential to be specific, scalable, and economical to produce. These salient features makes our BCAs attractive as a new class of reagents for use in array technology for understanding the regulation of kinase phosphorylation, which plays a central role in numerous signaling networks that are perturbed in human diseases, especially cancer. PUBLIC HEALTH RELEVANCE: Protein-kinases lay at the heart of signal transduction cascades, which when perturbed lead to human diseases such as cancer. Thus, the identification of new classes of bivalent reagents for capturing kinases in microarray applications has direct utility in understanding the pathophysiology of human diseases and aid in the development of novel diagnostics and therapeutics.