Discovery of Inhibitors for Small C-terminal Domain Phosphatases

描述(申请人提供)：人小C末端结构域(SCP)磷酸酶是最近发现的调节RNA聚合酶II C末端结构域的磷酸化状态的酶。更重要的是，SCP磷酸酶还被证明能抑制神经干细胞的不适当分化。在人类神经干细胞和非神经组织中，SCPS与REST/NRSF神经元沉默复合体密切相关，该复合体具有抑制神经基因转录的功能。因此，抑制SCPS磷酸酶活性的化合物可以成为指导神经发生和促进神经元再生的有力工具。一个长期的目标是开发一种治疗剂，通过抑制SCP磷酸酶，促进神经元分化，修复神经系统损伤。这些化合物最终将使患有神经元退行性疾病和神经元损伤的患者受益。在这项建议中，我们将使用孔雀石绿磷酸酶的天然底物来确定有效和特定的化学试剂来灭活孔雀石绿磷酸酶。将使用另外两种分析方法--荧光各向异性分析和基于细胞的转录分析来确认命中结果。确定的HITS将用于X-射线结构分析，这将为化合物优化提供必要的信息，并帮助我们提高抑制剂的选择性和亲和力。 与公共卫生相关：该项目的目的是开发一种人类SCPS的抑制剂。抑制SCPS可诱导干细胞向神经元分化，并诱导神经元基因表达。这种抑制物将是研究神经系统发育的重要试剂，因为它可能被用来分化内源性成体神经干细胞，以取代和修复神经元损伤，特别是在海马体和脊髓。临床目标是开发一种治疗退行性神经系统疾病的新疗法。

DESCRIPTION (provided by applicant): Human Small C-terminal Domain (Scp) phosphatases are recently identified enzymes that regulate the phosphorylation states of the C-terminal domain of RNA polymerase II. More importantly, the Scp phosphatases have also been shown to inhibit the inappropriate differentiation of neuronal stem cells. In human neuronal stem cells and non-neuronal tissues, Scps strongly associate with the REST/NRSF neuronal silencing complex that functions to inhibit transcription of neuronal genes. Therefore, chemical compounds inhibiting the phosphatase activity of Scps can be powerful tools to direct neurogenesis and promote the regeneration of neurons. A long-term goal is development of a therapeutic agent that, by inhibiting Scp phosphatases, would facilitate neuronal differentiation to repair nervous system damage. Such compounds would eventually benefit patients with neuron degenerative diseases and neuronal injuries. In this proposal, we would identify potent and specific chemical agents to inactivate Scp phosphatases in malachite green phosphatase assay using the natural substrates of the enzyme. The hits would be confirmed using another two assays, fluorescence anisotropy and cell-based transcription assays. The identified hits will be used in x-ray structural analysis which will provide essential information for compound optimization and help us to improve the selectivity and affinity of inhibitors. PUBLIC HEALTH RELEVANCE: The aim of this project is to develop an inhibitor of Human Scps. Inhibition of Scps leads to neuronal differentiation from stem cells and expression of neuronal genes. Such an inhibitor will be an important reagent for studying nervous system development, as it may be used to differentiate endogenous adult neuronal stem cells to replace and repair neuronal damage, especially in the hippocampus and spinal cord. The clinical goal is to develop a novel therapy for degenerative nervous system diseases.