Discovery of Inhibitors for Small C-terminal Domain Phosphatases

小 C 端结构域磷酸酶抑制剂的发现

基本信息

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
项目类别：
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

项目摘要

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）：人类小C末端结构域（SCP）磷酸酶最近被鉴定出调节RNA聚合酶II的C末端结构域的磷酸化状态的酶。更重要的是，SCP磷酸酶也已被证明可以抑制神经元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，SCP与其余/NRSF神经元沉默复合物紧密相关，可抑制神经元基因的转录。因此，抑制SCP的磷酸酶活性的化合物可以是指导神经发生和促进神经元再生的强大工具。一个长期目标是开发治疗剂，通过抑制SCP磷酸酶，它将促进神经元分化以修复神经系统的损害。这种化合物最终将使神经元退行性疾病和神经元损伤患者受益。在此提案中，我们将使用酶的天然底物确定有效和特定的化学剂在孔雀石绿色磷酸酶测定中灭活SCP磷酸酶。命中将使用另外两个测定法，荧光各向异性和基于细胞的转录测定法。确定的命中将用于X射线结构分析，该分析将为复合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SCP的抑制剂。 SCP的抑制导致与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，因为它可用于区分内源性成年神经元干细胞以替代和修复神经元损伤，尤其是在海马和脊髓中。临床目标是开发一种新的治疗神经系统疾病的疗法。