Chemical manipulation of creatine kinases to treat acute myeloid leukemia

项目总结： 在过去的十年里，许多癌症的基本代谢依赖性已经被揭示。这些 依赖性研究表明，某些新陈代谢节点是诱人的致癌药物靶点。 细胞死亡。在大多数情况下，致癌代谢蛋白还没有被下药，而且许多缺乏明显的 用于药理操作的捆绑口袋。在未吸毒的最突出的例子中 代谢性癌症的靶点是肌酸激酶(CKs)。CKs对肿瘤的生长和转移是必不可少的 多种癌症，尤其是侵袭性急性髓系白血病(AML)。CKS对 侵袭性AMLS是独特的，因为体细胞组织不依赖于CKS的生存能力。然而，尽管身为一名 高度可操作的药物靶点在AML中，不存在针对Cks的有效抑制剂。 我们最近开发了一种质谱学(MS)平台，允许快速筛选小分子 蛋白质组中与蛋白质半胱氨酸共价结合的分子。根据初步数据，我们 我把这个平台和一个小分子筛选库结合起来，系统地识别药物先导 将半胱氨酸定位在未下药的蛋白质上。在这样做的过程中，我们已经确定了一种有效的铅脚手架 通过选择性地针对一个关键的活性部位半胱氨酸残基，抑制CK酶家族。 此外，在高纳米分子浓度下，这种CK抑制剂对AML癌症具有选择性的细胞毒作用。 依靠CKS。本项目的目标是开发这种新型支架类的CK抑制剂，用于临床 在急性髓系白血病癌症中的应用。我们将检验这样一种假设，即这种支架的合理发展将会改善 通过靶向CK活性部位口袋的关键相互作用来提高效力和AML毒性。此外，我们还将 确定这种新的CK抑制剂类别在体内对AML小鼠模型是否有效。 在目标1中，我们将结合分子模拟方法和我们的初步合成孔径雷达数据来合理开发 一系列分子系统地探测对CK活性部位口袋的抑制效力。活动 这些分子的选择性将使用我们的MS平台、分离的CK动力学和 细胞性急性髓系白血病模型。同时，使用患者来源的急性髓系白血病细胞模型，其中CK是必不可少的，我们 将确定临床上相关的疾病改善产出，包括生存能力、集落形成和目标 毒性。在目标2中，我们将确定我们的CK抑制剂化疗类型在小鼠模型中的治疗效果 主要依赖CKS的EVI1阳性AML。总而言之，这个项目将推进一流的 CKS的抑制剂，这是AML癌症的一种未用药的代谢依赖。成功完成 这些目的将为AML的治疗定位这种新的化学类型，并为 了解CKS在急性髓系白血病和细胞代谢中的作用。

PROJECT SUMMARY: Over the last decade, essential metabolic dependencies of many cancers have been revealed. These dependencies have highlighted that certain nodes of metabolism are attractive drug targets to drive cancer cell death. For the most part, oncogenic metabolic proteins have not been drugged, and many lack obvious binding pockets for pharmacological manipulation. Among the most prominent examples of un-drugged metabolic cancer targets are creatine kinases (CKs). CKs are essential for growth and metastasis of numerous cancers, especially aggressive acute myeloid leukemias (AML). The essentiality of CKs for aggressive AMLs is distinct, as somatic tissues do not rely on CKs for viability. However, despite being a highly actionable drug target in AML, no potent inhibitors exist against CKs. We recently developed a mass spectrometric (MS) platform that allows for rapid screening of small molecules for covalent engagement with protein cysteines across the proteome. With preliminary data, we have combined this platform with a small molecule screening library to systematically identify drug leads targeting cysteines on un-drugged proteins. In doing so, we have identified a lead scaffold that is potently inhibitory against the CK family of enzymes, by selectively targeting a key active site cysteine residue. Moreover, at high nanomolar concentrations, this CK inhibitor is selectively cytotoxic to AML cancers that depend on CKs. The goal of this project is to develop this new scaffold class of CK inhibitor for clinical application in AML cancers. We will test the hypothesis that rational development of this scaffold will improve potency and AML toxicity by targeting key interactions at the CK active site pocket. Moreover, we will determine whether this new CK inhibitor class is an effective therapeutic in a mouse model of AML in vivo. In Aim 1 we will combine molecular modeling approaches and our preliminary SAR data to rationally develop a series of molecules to systematically probe inhibitory potency against the CK active site pocket. Activity and selectivity of these molecules will be determined using our MS platform, isolated CK kinetics, and cellular AML models. In parallel, using patient-derived cellular models of AML, for which CK is essential, we will determine clinically relevant disease-modifying outputs, including viability, colony forming, and on-target toxicity. In Aim 2 we will determine therapeutic efficacy of our CK inhibitor chemotype in mouse models EVI1-positive AML that rely essentially on CKs. Taken together, this project will advance a first-in-class inhibitor of CKs, which is an un-drugged metabolic dependency of AML cancers. Successful completion of these Aims would position this new chemotype for treatment of AML, and provide a new chemical probe for understanding the role CKs play in AML and cellular metabolism.