Chemical manipulation of creatine kinases to treat acute myeloid leukemia

项目概要： 在过去的十年中，许多癌症的基本代谢依赖性已经被揭示。这些 依赖性突出了某些代谢节点是驱动癌症的有吸引力的药物靶点， 细胞死亡在大多数情况下，致癌代谢蛋白没有被药物，许多缺乏明显的 用于药理学操作的结合口袋。在最突出的例子中， 代谢性癌症靶点是肌酸激酶（CK）。CKs对肿瘤的生长和转移至关重要， 许多癌症，特别是侵袭性急性髓性白血病（AML）。CKs的重要性 侵袭性AML是独特的，因为体细胞组织不依赖CK生存。然而，尽管是一个 在AML中高度可操作的药物靶向，没有针对CK的有效抑制剂存在。 我们最近开发了一种质谱（MS）平台，可以快速筛选小分子 分子与蛋白质组中的蛋白质半胱氨酸共价结合。根据初步数据，我们 我将这个平台与小分子筛选库相结合，系统地识别药物先导物， 将半胱氨酸定位在未加药物的蛋白质上。在这样做的过程中，我们已经确定了一个铅支架， 通过选择性靶向关键活性位点半胱氨酸残基抑制CK家族酶。 此外，在高纳摩尔浓度下，这种CK抑制剂对AML癌症具有选择性细胞毒性， 依赖CK。本项目的目标是开发这种新型的CK抑制剂支架，用于临床 在AML癌症中的应用我们将测试这一假设，即合理开发这种支架将改善 通过靶向CK活性位点口袋处的关键相互作用来降低效价和AML毒性。而且还要 确定这种新的CK抑制剂类是否是体内AML小鼠模型中的有效治疗剂。 在目标1中，我们将结合联合收割机分子模拟方法和我们的初步SAR数据， 一系列分子系统地探测对CK活性位点口袋的抑制效力。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， 细胞AML模型。与此同时，使用患者来源的AML细胞模型，CK是必不可少的， 将确定临床相关的疾病改善输出，包括活力、集落形成和靶向 毒性在目标2中，我们将确定我们的CK抑制剂化学型在小鼠模型中的治疗功效 EVI1阳性AML主要依赖于CK。总的来说，这个项目将推动一流的 CKs抑制剂，这是AML癌症的非药物代谢依赖性。成功完成 这些目标将定位这种新的化学型用于治疗AML，并提供一种新的化学探针， 了解CKs在AML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.