Development of small molecule inhibitors of protein kinase D

描述（由申请人提供）：蛋白激酶D （PKD）是一个新的关键第二信使二酰基甘油靶点家族，可以被g蛋白偶联受体激动剂和生长因子激活。PKD家族- PKD1, 2和3 -在许多基本细胞过程中起着重要作用。PKD的失调与多种病理状况和癌症有关。然而，缺乏pkd特异性抑制剂严重阻碍了我们理解pkd特异性信号通路和生物学功能，并将其作为人类疾病的靶点。我们最近报道了首个有效的选择性PKD小分子抑制剂：苯并唑氮匹诺酮CID755673，通过对196,173个化合物的高通量筛选筛选出。CID755673是一种亚微摩尔效价的泛pkd抑制剂。它具有细胞活性，阻断PKD已知的生物学作用，抑制前列腺癌细胞的增殖、迁移和侵袭等与癌症相关的特性。重要的是，与至少20种不同的激酶（包括CAMKII1、AKT、PKA和几种PKC异构体）相比，该抑制剂在酶抑制方面不与ATP竞争，对PKD具有高度选择性。最关键的是，CID755673的结构优化得到了一种新的苯并噻吩衍生物kb-NB142-70，具有低纳摩尔效价和更高的PKD选择性。我们假设CID755673可以被优化为一类具有独特作用机制的新型PKD抑制剂，可以有效和选择性地阻断PKD功能。我们的长期目标是建立一种系统的方法来开发cid755673衍生的探针分子作为创新的PKD抑制剂。我们提出的研究策略主要集中在CID755673及其类似物的先导物优化、机制评价和体内疗效评估。提出了三个具体目标，代表了开发强效和选择性PKD药理学消融剂的第一个主要努力：目标1。通过结构修饰优化CID755673，获得更有效和选择性的衍生物。目标2。确定CID755673及其类似物对PKD的精细选择性的分子机制。目标3。验证CID755673及其衍生物在体内导致PKD功能有效和选择性阻断的假设。

DESCRIPTION (provided by applicant): Protein kinase D (PKD) is a novel family of the key second messenger diacylglycerol targets that can be activated by G-protein coupled receptor agonists and growth factors. The family of PKD - PKD1, 2 and 3 - plays important roles in many fundamental cellular processes. Deregulation of PKD has been implicated in multiple pathological conditions and cancer. However, the lack of a PKD-specific inhibitor has severely impeded our ability to understand PKD-specific signaling pathways and biological functions and to target it in human diseases. We have recently reported the first potent and selective small molecule inhibitor for PKD: benzoxoloazepinolone CID755673 that was identified through a high throughput screening campaign of 196,173 compounds. CID755673 is a pan-PKD inhibitor with submicromolar potencies. It was cell active and blocked the known biological actions of PKD and suppressed cancer-associated properties of proliferation, migration and invasion of prostate cancer cells. Importantly, this inhibitor was not competitive with ATP for enzyme inhibition and was highly selective for PKD when compared to at least 20 different kinases including CAMKII1, AKT, PKA and several PKC isoforms. Most critically, structural optimization of CID755673 has yielded a novel benzthiophene derivative kb-NB142-70 with low nanomolar potency and greater selectivity for PKD. We hypothesize that CID755673 can be optimized to a class of novel PKD inhibitors with unique mechanisms of action for potent and selective blockade of PKD functionality. Our long term goal is to establish a systemic approach to develop CID755673-derived probe molecules as innovative PKD inhibitors. Our proposed research strategies focus on lead optimization, mechanistic evaluation and in vivo efficacy assessment of CID755673 and its analogs. Three specific aims are proposed, representing the first major effort towards the development of potent and selective pharmacological ablative agents for PKD: Aim 1. Optimize CID755673 by structural modifications to access more potent and selective derivatives. Aim 2. Determine the molecular mechanisms underlying the exquisite selectivity of CID755673 and its analogs for PKD. Aim 3. Test the hypothesis that CID755673 and derivatives cause potent and selective blockade of PKD functionality in vivo. PUBLIC HEALTH RELEVANCE: The protein kinase D family is a novel receptor of the key second messenger diacylglycerol and an emerging therapeutic target for cancer and other diseases. Further understanding the role of PKD in biological processes and targeting it in human diseases have been severely impeded by the lack of a PKD-specific inhibitor that can be readily applied to cells and in animals. We have recently discovered the first potent and selective cell-active small molecule inhibitor for PKD - CID755673. We hypothesize that CID755673 can be optimized to generate a class of novel PKD inhibitors with unique mechanisms of action for potent and selective blockade of PKD functionality. The proposed studies focus on lead optimization, mechanistic evaluation and in vivo efficacy assessment of these novel chemical entities. The successful completion of the study will lead to a novel class of PKD inhibitors that can be used as powerful tools for dissecting PKD-mediated signaling pathways and functionalities in cells and animals. It will also provide the foundation for potential future clinical development of the compounds as novel antitumor agents.