Structure-based Identification and Functional Characterization of SSH1 Inhibitors

SSH1 抑制剂的基于结构的鉴定和功能表征

• 批准号: 8872959
• 负责人: Yu Chen
• 金额: $ 22.43万
• 依托单位: UNIVERSITY OF SOUTH FLORIDA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8872959
• 关键词: APP-PS1; Actins; Alzheimer' s Disease; Amyloid; Amyloid beta-Protein; Apoptosis; Biological; Biological Assay; Brain; Catalytic Domain; Cells; Characteristics; Chemicals; Complex; Cysteine; Docking; Drug Kinetics; Exhibits; F-Actin; Feedback; Future; Impaired cognition; Impairment; In Vitro; Induction of Apoptosis; LIMK1 gene; Lead; Ligand Binding; Ligands; Link; Mitochondria; Modeling; Modification; Molecular; Mus; Oxidative Stress; Pathology; Pathway interactions; Patients; Phosphoric Monoester Hydrolases; Phosphorylation; Production; Protein Dephosphorylation; Proteins; Roentgen Rays; Scaffolding Protein; Signal Transduction; Site-Directed Mutagenesis; Specificity; Structural Models; Structure; Structure-Activity Relationship; Superoxides; Surface; Synapses; Therapeutic; abeta accumulation; base; cofilin; cofilin 2; design; functional group; improved; inhibitor/antagonist; mitochondrial dysfunction; neurotoxic; neurotoxicity; novel; prevent; public health relevance; research study; retinal rods; screening; tau Proteins; tau dysfunction; therapeutic target; virtual

 DESCRIPTION (provided by applicant): As the major defining characteristic of Alzheimer's disease (AD) brains is the excessive accumulation of toxic proteins called Aβ and Tau, understanding the biological mechanisms by which Aβ connects to tau dysfunction are critical for designing effective therapeutic treatments for AD. We have identified SSH1, a phosphatase that activates Cofilin, as a promising therapeutic target for AD, as SSH1 impacts both Aβ and Tau. We have also identified several SSH1 inhibitor compounds that effectively decrease both Aβ-induced neurotoxicity and Tau hyperphosphorylation while decreasing Aβ production. In the proposal, we will perform chemical modifications of SSH1 inhibitor compounds to verify the functional groups critical for SSH1 inhibition and establish a platform for lead optimization. We will optimize and carry out in vitro phosphatase activity assays for SSH1 and other dual specificity phosphatases. We will also perform AD-relevant cell-based assays using existing and chemically modified compounds. Complex X-ray crystal structures will be determined using existing and modified compounds in physical contact with the catalytic domain of SSH1, to investigate the molecular basis of their interactions. Double perturbation experiments will be used to probe the contribution of specific inhibitor functional groups and protein residues to ligand binding using modified compounds and site-specific mutagenesis of SSH1. Based on a revised structural model, virtual docking and structure-based design will be performed to improve existing ligand efficiency, and new virtual screening will be carried out to identify additional SSH1 inhibitor chemotypes. Finally, based on the new structure-based ligand efficiency model, we will perform synthesis of a modified compound to demonstrate the proof-of-concept that such modified compound exhibits improved SSH1 inhibitory activity and ADME characteristics in vitro. Therefore, results of this R21 proposal are expected to yield valuable structure function relationship between SSH1 and existing and new compounds as well as establish a structure-based platform for extensive lead optimization and pharmacokinetic studies in a future R01 proposal.

 描述（由申请人提供）：由于阿尔茨海默病（AD）大脑的主要定义特征是称为Aβ和Tau的毒性蛋白质的过度积累，因此了解Aβ与tau功能障碍相关的生物学机制对于设计有效的AD治疗方法至关重要。我们已经鉴定了SSH1，一种激活Cofilin的磷酸酶，作为AD的有希望的治疗靶点，因为SSH1影响Aβ和Tau。我们还鉴定了几种SSH1抑制剂化合物，其有效降低Aβ诱导的神经毒性和Tau过度磷酸化，同时降低Aβ产生。在该提案中，我们将对SSH1抑制剂化合物进行化学修饰，以验证对SSH1抑制至关重要的官能团，并建立铅优化平台。我们将优化并进行SSH1和其他双特异性磷酸酶的体外磷酸酶活性测定。我们还将使用现有的和化学修饰的化合物进行AD相关的基于细胞的测定。复杂的X射线晶体结构将使用与SSH1的催化结构域物理接触的现有和改性化合物来确定，以研究它们相互作用的分子基础。双扰动实验将用于探测特定抑制剂官能团和蛋白质残基对使用修饰的化合物和SSH1的位点特异性诱变的配体结合的贡献。基于修订的结构模型，将进行虚拟对接和基于结构的设计，以提高现有配体的效率，并将进行新的虚拟筛选，以确定额外的SSH1抑制剂化学型。最后，基于新的基于结构的配体效率模型，我们将进行修饰化合物的合成，以证明这种修饰化合物在体外表现出改善的SSH1抑制活性和ADME特性的概念验证。因此，该R21提案的结果有望在SSH1与现有和新化合物之间产生有价值的结构功能关系，并在未来的R01提案中为广泛的先导化合物优化和药代动力学研究建立基于结构的平台。