Chemical Probes as Allosteric Modulators of CK2 Alpha Prime Targeting Huntington's Disease

项目总结 CK2α‘是人蛋白激酶CK2全酶的两个催化亚基之一，在 亨廷顿病(HD)的细胞和动物模型，以及患有HD的人类患者。目前没有 CK2α‘的选择性抑制剂可用。我们的工作表明，CK2α‘参与了蛋白的过度磷酸化和 应激保护热休克转录因子1(HSF1)的降解HSF1有几个保护作用 在体内，包括应激保护性伴侣和突触蛋白的调节，以及能量代谢。HSF1 在缺乏CK2α‘(ZQ175HD)等位基因的HD小鼠模型中，水平升高，导致增加 伴侣蛋白表达和兴奋性突触密度，减少HTT聚集和炎症，以及 改善了运动行为。鉴于这些令人兴奋和有希望的结果，我们正在启动一项计划，以确定 CK2α‘的选择性变构抑制剂可作为体外和体内靶向验证的化学探针 学习。这一探索性项目的唯一具体目标是鉴定和表征变构抑制剂。 CK2α‘，可作为选择性探针开发的先导。在此，我们建议采用ADP-GloTM ChemDiv变构酶抑制剂(CDAKI)文库的高通量发光筛选，增加了 我们将发现一种与CK2α‘变构结合的小分子的可能性。活性化合物将是 用等温滴定量热法(ITC)和X射线结晶学对其进行了进一步表征。已确认活动 化合物将通过商业使用构效关系(SAR)进行验证。我们的工作假设是 这个已知的变构酶抑制剂的文库将为结构使能的 化合物的发展导致选择性变构探针。这项工程对人类的潜在影响 健康是相当重要的。有一种尚未得到满足的医疗需求，即可以阻止或逆转 与HD相关的认知和运动能力下降。这项工作将对实地产生积极影响，因为它将提供 为验证治疗开发的新靶点而走向化学探针的途径。最终的 α‘的选择性变构抑制剂的开发将解决这一未得到满足的医学需求，并代表着一种 高清领域的重大进步。

PROJECT SUMMARY CK2α’, one of two catalytic subunits of human protein kinase CK2 holoenzyme, is inappropriately upregulated in cellular and animal models of Huntington’s disease (HD), and in human patients with HD. There are currently no selective inhibitors for CK2α’ available. Our work shows that CK2α’ is involved in the hyperphosphorylation and degradation of the stress protective Heat Shock transcription Factor 1 (HSF1). HSF1 has several protective roles in vivo, including regulation of stress protective chaperones and synaptic proteins, and energy metabolism. HSF1 levels are increased in an HD mouse model lacking one allele of CK2α’ (zQ175 HD), leading to increased chaperone expression and excitatory synapse density, decreased HTT aggregates and inflammation, and improved motor behavior. Given these exciting and promising results, we are initiating a program to identify selective allosteric inhibitors of CK2α’ that can serve as chemical probes for in vitro and in vivo target validation studies. The single specific aim of this exploratory project is to identify and characterize allosteric inhibitors of CK2α’ that can serve as leads for selective probe development. Herein, we propose to employ an ADP-GloTM luminescence high-throughput screen of the ChemDiv Allosteric Kinase Inhibitor (CDAKI) Library, increasing the likelihood that we will discover a small-molecule that binds allosterically to CK2α’. Active compounds will be further characterized by isothermal titration calorimetry (ITC) and x-ray crystallography. Confirmed active compounds will be validated using SAR (structure-activity relationship) by commerce. Our working hypothesis is that this library of known allosteric kinase inhibitors will generate excellent starting points for structurally-enabled compound development leading to selective allosteric probes. The potential impact of this project on human health is considerable. There is an unmet medical need for therapeutic agents that can halt or reverse the cognitive and motor decline associated with HD. This work will have a positive impact on the field as it will provide a path toward chemical probes for the validation of a new target for therapeutic development. The eventual development of a selective allosteric inhibitor of CK2α’ would address this unmet medical need and represent a significant advancement in the field of HD.