Designed Inhibitors of Pin1 in Mitosis

有丝分裂中 Pin1 的设计抑制剂

• 批准号: 7477724
• 负责人: FELICIA A ETZKORN
• 金额: $ 18.18万
• 依托单位: VIRGINIA POLYTECHNIC INST AND ST UNIV
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7477724
• 关键词: Active Sites; Affinity; Alzheimer' s Disease; Amides; Binding; Biological; Biological Assay; Biological Availability; Biological Process; C-terminal; Catalysis; Catalytic Domain; Cell Cycle; Cell Cycle Checkpoint; Cell Cycle Progression; Cell Cycle Proteins; Cell Cycle Regulation; Cell Membrane Permeability; Cells; Collaborations; Cyclin D1; Development; Dipeptides; Disease; Docking; Enzyme-Linked Immunosorbent Assay; Enzymes; Esters; Goals; Induction of Apoptosis; JUN gene; Ketones; Lead; Length; Libraries; Ligands; Masks; Measures; Mitosis; Mitotic; Modeling; Modification; Molecular; NMR Spectroscopy; Numbers; Object Attachment; Peptides; Peptidylprolyl Isomerase; Permeability; Pharmaceutical Preparations; Phase; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphoserine; Phosphotransferases; Play; Prodrugs; Proline; Range; Regulation; Research Personnel; Roentgen Rays; Role; Screening procedure; Signal Transduction; Solid; Structural Models; Structure; Testing; Threonine; Work; X-Ray Crystallography; analog; base; cdc25 Phosphatase; comparative; design; ginsenoside M1; improved; in vivo; inhibitor/antagonist; inorganic phosphate; interest; jun Oncogene; programs; tau Proteins; tool

DESCRIPTION (provided by applicant): Pin1 is an enzyme that catalyzes isomerization of phosphoserine/threonine-proline (pSer/Thr-Pro) amides. Pin1 is hypothesized to regulate signal transduction leading to mitosis by a conformational switch, isomerizing specific prolyl amides in cell cycle regulatory proteins. Pin1 has also been shown to regulate the activity of Alzheimer's associated tau protein. The unique peptidyl-prolyl isomerase (PPIase) enzymatic activity of Pin1 in the regulation of biological processes is interesting for fundamental reasons. Pin1 consists of two domains that both bind pSer/Thr-Pro motifs with distinct selectivity's. Specific ligands will be developed for each domain separately to be used as tools to study regulation by Pin1. In the first Specific Aim, we propose the design, parallel synthesis and assay of libraries of conformationally constrained trans- proline mimics targeted to the Pin1 WW domain. Parallel synthesis will use solid phase attachment through the invariant phosphate, and amide bond couplings to produce compounds of sufficient purity for screening. A high-throughput competitive enzyme-linked immunosorbent assay (ELISA) will be developed to assay WW domain binding. The WW domain ligands will also be tested for noncompetitive inhibition in the catalytic assay because we have shown inhibition of catalysis by a peptide binding to the WW domain. In the second Aim, libraries will be derived from compounds targeted to the catalytic domain that we have already synthesized: pSer-c/s-Pro isostere, reduced amide, ketone and alpha-ketoamide motifs. The catalytic domain inhibitors will be screened in a high-throughput enzymatic assay, and the mode of inhibition will be determined for the best inhibitors. In the third Aim, the two types of ligands for Pint will be combined into bivalent inhibitors to study the structural and dynamic interactions between the two domains of Pin1. The fourth Aim concerns development of phosphate prodrugs, masked to improve membrane permeability. We will also develop diflurophosphonates stabilized towards phosphatases. These two modifications will allow these compounds to be used in whole cells and in vivo. The best ligands will be useful in collaborations designed to elucidate the structure, function and dynamic relationships within Pin1 and between Pin1 and its ligands.

描述（由申请人提供）：Pin1是一种催化磷酸丝氨酸/苏氨酸-脯氨酸（pSer/Thr-Pro）酰胺异构化的酶。据推测，Pin1 通过构象转换调节导致有丝分裂的信号转导，使细胞周期调节蛋白中的特定脯氨酰胺异构化。 Pin1 还被证明可以调节阿尔茨海默病相关 tau 蛋白的活性。 Pin1 在生物过程调节中独特的肽基脯氨酰异构酶 (PPIase) 酶活性出于根本原因而令人感兴趣。 Pin1 由两个结构域组成，这两个结构域均以不同的选择性结合 pSer/Thr-Pro 基序。将为每个结构域分别开发特定配体，用作研究 Pin1 调节的工具。在第一个具体目标中，我们提出了针对 Pin1 WW 结构域的构象受限反式脯氨酸模拟物文库的设计、并行合成和测定。平行合成将通过不变的磷酸盐和酰胺键偶联使用固相连接来产生足够纯度的化合物用于筛选。将开发高通量竞争性酶联免疫吸附测定 (ELISA) 来测定 WW 结构域结合。 WW 结构域配体还将在催化测定中测试非竞争性抑制，因为我们已经显示了与 WW 结构域结合的肽对催化的抑制。在第二个目标中，文库将从针对我们已经合成的催化结构域的化合物衍生：pSer-c/s-Pro等排体、还原酰胺、酮和α-酮酰胺基序。将通过高通量酶测定筛选催化域抑制剂，并确定最佳抑制剂的抑制模式。在第三个目标中，Pint 的两种配体将被组合成二价抑制剂，以研究 Pin1 的两个结构域之间的结构和动态相互作用。第四个目标涉及磷酸盐前药的开发，以提高膜的渗透性。我们还将开发对磷酸酶稳定的二氟膦酸盐。这两种修饰将使这些化合物能够用于全细胞和体内。最好的配体将有助于旨在阐明 Pin1 内部以及 Pin1 与其配体之间的结构、功能和动态关系的合作。