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

项目摘要 人蛋白激酶CK2全酶的两个催化亚基之一，CK2 β '在人细胞中不适当地上调。 亨廷顿氏病（HD）的细胞和动物模型，以及患有HD的人类患者。目前没有 选择性CK2 α抑制剂。我们的工作表明，CK2 α '参与了过度磷酸化， 应激保护性热休克转录因子1（HSF 1）的降解。HSF1具有多种保护作用 在体内，包括调节应激保护分子伴侣和突触蛋白，以及能量代谢。HSF1 在缺乏CK2 α '（zQ175 HD）的一个等位基因的HD小鼠模型中， 伴侣蛋白表达和兴奋性突触密度，减少HTT聚集和炎症， 改善运动行为。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， CK2 α '的选择性变构抑制剂，可用作体外和体内靶点验证的化学探针 问题研究该探索性项目的单一具体目标是鉴定和表征以下的变构抑制剂： CK2 α '可作为选择性探针开发的先导。在此，我们提出采用ADP-GloTM 发光高通量筛选ChemDiv变构激酶抑制剂（CDAKI）文库，增加 我们将发现一种与CK2 β '变构结合的小分子的可能性。活性化合物将 进一步通过等温滴定量热法（ITC）和X射线晶体学表征。确认激活 化合物将通过商业使用SAR（结构-活性关系）进行验证。我们的假设是 这种已知的变构激酶抑制剂的文库将为结构激活的 化合物的开发导致选择性变构探针。该项目对人类的潜在影响 健康是相当可观的。存在对可以停止或逆转肿瘤的治疗剂的未满足的医学需求。 与HD相关的认知和运动能力下降。这项工作将对外地产生积极影响，因为它将提供 一条通往化学探针的道路，用于验证治疗开发的新靶点。最终 开发选择性CK2 α '变构抑制剂将解决这一未满足的医疗需求， 在HD领域取得了重大进展。

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.