Discovery of Inhibitors for Small C-terminal Domain Phosphatases

描述（由申请人提供）：人类小c端结构域（Scp）磷酸酶是最近发现的调节RNA聚合酶II c端结构域磷酸化状态的酶。更重要的是，Scp磷酸酶也被证明可以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神经元沉默复合物密切相关，该复合物具有抑制神经元基因转录的功能。因此，抑制Scps磷酸酶活性的化合物可能是指导神经发生和促进神经元再生的有力工具。长期目标是开发一种治疗剂，通过抑制Scp磷酸酶，促进神经元分化以修复神经系统损伤。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。在这项提议中，我们将利用孔雀石绿磷酸酶的天然底物，在孔雀石绿磷酸酶测定中确定有效和特异性的化学制剂来灭活Scp磷酸酶。这些命中将使用另外两种分析，荧光各向异性和基于细胞的转录分析来证实。确定的命中点将用于x射线结构分析，这将为化合物优化提供重要信息，并帮助我们提高抑制剂的选择性和亲和力。

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.