Development and Application of Selective Covalent Cdk7 Inhibitors

摘要：选择性蛋白激酶抑制剂是研究细胞信号通路的有力工具。 以及验证治疗各种癌症的药物靶点。现有的药物发现范式 今天首先是在学术机构中鉴定和确认激酶靶标，然后是 由制药部门开发的抑制剂。这种模式的一个主要缺点是严重的 缺乏针对大多数激酶的选择性抑制剂阻碍了最初的药理学概念验证研究和 因此，不鼓励制药部门进一步探索这些目标。我们的实验室有 试图通过开发发现第一类激酶的有效方法来解决这一不足 抑制剂，然后被用作药理工具来研究其功能和潜在的治疗作用 有问题的激酶的相关性。在过去的五年里，我们的实验室得到了广泛的发展 分布了ALK、Mps1、ERK5、mTOR、LRRK2、FGFRs、JNKs和T790M突变形式的第一批抑制剂 EGFR；其中几种是目前的标准参考化合物。 在这项申请中，我们组建了一个多学科团队，将药物化学 (Nathanael Gray)、结构生物学(Jane Endicott)、转录(Richard Young)和癌症生物学 翻译研究(康斯坦丁·米西亚兹)，将使第一个 CDK7的高度有效和选择性的抑制剂。细胞周期蛋白依赖性蛋白激酶是一类高度保守的 由20个成员(CDK1-20)组成的蛋白激酶，与29个调节蛋白家族相关 被称为旋转体。它们调节大量的细胞功能，包括细胞周期(CDK1-6)、转录 (CDK7-13、19)和剪接(CDK11)。而调节细胞周期的CDK已经收到了大量的 作为药物发现的目标，所谓的转录CDK(tCDKs-CDK7-13，19)已经引起了人们的关注 收到的要少得多。我们发现了一种前所未有的方法来开发选择性CDK7抑制剂，方法是 共价靶向位于激活域之外的独特的半胱氨酸(Cys)残基。我们已经获得了 CDK7抑制剂在大量癌症中的活性的令人信服的初步数据，但在这里我们 建议重点研究靶向CDK7治疗多发性骨髓瘤的可能性。 浆细胞瘤，过度转录是疾病的标志，根据我们的初步数据， CDK7起着重要的病理生理作用。这将通过一种专注的药物化学来实现 运动(目标1)和由详细的机械特性指导(目标2)，随后是临床前 多发性骨髓瘤细胞模型和小鼠模型的评价(目标3)。

Abstract: Selective protein kinase inhibitors are powerful tools for interrogating cellular signaling pathways and validating drug targets for the treatment of a variety of cancers. The drug discovery paradigm that exists today starts with the identification and validation of kinase targets primarily at academic institutions followed by inhibitor development by the pharmaceutical sector. A major shortcoming of this paradigm is the severe shortage of selective inhibitors for most kinases hampers initial pharmacological proof-of-concept studies and therefore discourages further exploration of these targets by the pharmaceutical sector. Our laboratory has attempted to address this deficiency by developing efficient approaches to the discovery of first-in-class kinase inhibitors, which are then used as pharmacological 'tools' to investigate the functions and potential therapeutic relevance of the kinase in question. Over the last five years, our laboratory has developed and widely distributed the first inhibitors of ALK, Mps1, Erk5, mTor, LRRK2, FGFRs, JNKs, and the T790M mutant form of EGFR; several of which are currently standard reference compounds. In this application we have assembled a multidisciplinary team that integrates medicinal chemistry (Nathanael Gray), structural biology (Jane Endicott), transcription (Richard Young) and cancer biology and translational research (Constantine Mitsiades) that will enable the development and application of the first highly potent and selective inhibitors of CDK7. The cyclin-dependent kinases are a highly conserved class of protein kinases that consist of 20 members (CDK1-20) that associate with a family of 29 regulatory proteins called cyclins. They regulate a large number of cellular functions including cell cycle (CDK1-6), transcription (CDK7-13, 19), and splicing (CDK11). While the cell cycle regulating CDKs have received a significant amount of attention as drug discovery targets the so-called 'transcriptional CDKs' (tCDKs - CDK7-13, 19) have received much less. We have discovered an unprecedented means of developing selective CDK7 inhibitors by covalently targeting a unique cysteine (Cys) residue located outside of the kinase domain. We have obtained compelling preliminary data of the activity of CDK7 inhibitors in a large number of cancers, but here we propose to focus on investigating the potential of targeting CDK7 for the treatment of Multiple Myeloma, a plasma cell tumor where excess transcription is a hallmark of the disease and, based on our preliminary data, CDK7 plays a critical pathophysiological role. This will be accomplished through a focused medicinal chemistry campaign (Aim 1) and guided by detailed mechanistic characterization (Aim 2), followed by preclinical evaluation in cellular and murine models of Multiple Myeloma (Aim 3).