Discovery of Inhibitors for Small C-terminal Domain Phosphatases

小 C 端结构域磷酸酶抑制剂的发现

• 批准号: 8063541
• 负责人: Yan Jessie Zhang
• 金额: $ 3.69万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8063541
• 关键词: Active Sites; Adult; Affinity; Binding; Biological Assay; C-terminal; Cells; Chemical Agents; Chemicals; Clinical; Complex; Crystallography; Degenerative Disorder; Development; Dominant-Negative Mutation; Embryo; Enzyme Inhibitor Drugs; Enzyme Inhibitors; Enzymes; Exhibits; Family; Fluorescence Anisotropy; Genes; Genetic Transcription; Goals; Hippocampus (Brain); Human; Methods; MicroRNAs; Natural regeneration; Nervous System Trauma; Nervous system structure; Neurodegenerative Disorders; Neuronal Differentiation; Neuronal Injury; Neurons; Patients; Peptides; Phosphoric Monoester Hydrolases; Phosphorylation; Process; Proteins; RNA Polymerase II; Reagent; Specificity; Spinal Cord; Stem cells; Structure; Therapeutic Agents; Tissues; Transcription Initiation; base; design; enzyme substrate; high throughput screening; improved; inhibitor/antagonist; nervous system development; neurogenesis; neuron development; novel; premature; public health relevance; repaired; small molecule; structural biology; tool; transcription factor REST

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.

描述（由申请人提供）：人小C端结构域（Scp）磷酸酶是最近鉴定的调节RNA聚合酶II的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。在人类神经元干细胞和非神经元组织中，Scps与REST/NRSF神经元沉默复合物密切相关，该复合物的功能是抑制神经元基因的转录。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。长期目标是开发一种治疗剂，通过抑制Scp磷酸酶，将促进神经元分化以修复神经系统损伤。这些化合物最终将有益于患有神经元变性疾病和神经元损伤的患者。 在这个建议中，我们将确定有效的和特定的化学试剂，以消除SCp磷酸酶在孔雀石绿色磷酸酶测定使用的酶的天然底物。将使用另外两种测定法，荧光各向异性和基于细胞的转录测定法来确认命中。所识别的命中将用于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。 公共卫生相关性：该项目的目的是开发人类Scps的抑制剂。Scps的抑制导致干细胞的神经元分化和神经元基因的表达。这种抑制剂将是研究神经系统发育的重要试剂，因为它可用于分化内源性成体神经元干细胞以替代和修复神经元损伤，特别是在海马和脊髓中。临床目标是为退行性神经系统疾病开发一种新的治疗方法。

项目成果

Crystal structure of Ssu72, an essential eukaryotic phosphatase specific for the C-terminal domain of RNA polymerase II, in complex with a transition state analogue.

• DOI: 10.1042/bj20101471
• 发表时间: 2011-03
• 期刊: The Biochemical journal
• 作者: Yong Zhang;Mengmeng Zhang;Yan Zhang

Selective inactivation of a human neuronal silencing phosphatase by a small molecule inhibitor.

• DOI: 10.1021/cb100357t
• 发表时间: 2011-05-20
• 期刊: ACS chemical biology
• 影响因子: 4
• 作者: Zhang M;Cho EJ;Burstein G;Siegel D;Zhang Y
• 通讯作者: Zhang Y

Cross-talk of phosphorylation and prolyl isomerization of the C-terminal domain of RNA Polymerase II.

RNA 聚合酶 II C 末端结构域的磷酸化和脯氨酰异构化的串扰。

• DOI: 10.3390/molecules19021481
• 发表时间: 2014
• 期刊: Molecules (Basel, Switzerland)
• 作者: Yogesha,SD;Mayfield,JoshuaE;Zhang,Yan
• 通讯作者: Zhang,Yan

Viewing serine/threonine protein phosphatases through the eyes of drug designers.

从药物设计者的角度看丝氨酸/苏氨酸蛋白磷酸酶。

• DOI: 10.1111/febs.12481
• 发表时间: 2013-10
• 期刊: The FEBS journal
• 作者: Zhang M;Yogesha SD;Mayfield JE;Gill GN;Zhang Y
• 通讯作者: Zhang Y