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Small-molecule Inhibition of the Interactions of the Urokinase Receptor: A Targe

尿激酶受体相互作用的小分子抑制：一个目标

基本信息

批准号：
8233459
负责人：
Samy Meroueh
金额：
$ 24.8万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-05-15 至 2014-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8233459
关键词：
Adhesions Affinity Antineoplastic Agents Applications Grants Binding Biochemical Biological Assay Blood - brain barrier anatomy Breast Caco-2 Cells Calorimetry Cell Adhesion Cell Culture Techniques Cell Line Cell Proliferation Cell Surface Proteins Cell Surface Receptors Cell surface Cells Cellular biology Chemicals Clinic Clinical Pharmacology Co-Immunoprecipitations Collaborations Colon Adenocarcinoma Complement Complex Computational Biology Core Facility Crystallization Cytochrome P450 Data Development Docking Drug Design Drug Kinetics Drug Transport Ensure Environment Evaluation Event Exhibits Extracellular Matrix Degradation Fluorescence Polarization Free Energy Future G protein coupled receptor kinase Generations Genomics Glioblastoma H1299 Head Human Indiana Integrin Binding Integrins Interdisciplinary Study Knowledge Lead Letters Libraries Ligands Literature Lung Malignant Neoplasms Mammary Neoplasms Mediating Metabolism Molecular Mus Neoplasm Metastasis Penetration Peptide Hydrolases Permeability Pharmaceutical Preparations Pharmacologic Substance Phosphorylation Plasminogen Positioning Attribute Primary Neoplasm Process Property Proteins Proteolysis Proteome Reporting Research Rewards Risk Role Screening procedure Serine Protease Serum Albumin Signal Pathway Signal Transduction Signaling Protein Site Solutions Staging Statistical Models Structure Surface Plasmon Resonance Synthesis Chemistry System Systems Development Testing Therapeutic Titrations Toxic effect Transmembrane Domain Tumor Cell Invasion Tumor Cell Line U118 Urokinase Urokinase Plasminogen Activator Receptor Validation Work Xenograft procedure angiogenesis anti-cancer therapeutic base biophysical chemistry cell motility chemical property chemical synthesis cheminformatics combinatorial crosslink design dosage drug discovery expectation experience fibrosarcoma forging high throughput screening in vivo inhibitor/antagonist innovation insight medical schools migration molecular dynamics neglect neoplastic cell prevent professor programs protein protein interaction receptor receptor binding small molecule small molecule libraries structural biology success three dimensional structure tumor tumor xenograft tyrosine receptor virtual

项目摘要

PROJECT SUMMARY/ABSTRACT The urokinase receptor (uPAR) is a GPI-anchored protein that serves as a receptor to the serine protease urokinase (uPA). Upon docking to its receptor, uPA is activated, an event that leads to pericellular proteolysis and degradation of the extracellular matrix, a primary barrier between tumor and vasculature. But it is now widely recognized that while uPAR is not capable of signaling, the uPAR/uPA complex interacts and activates a number of cell surface receptors such as integrins, receptor tyrosine kinases, and GPCRs. The uPAR/uPA interaction has been implicated with nearly every step of tumor invasion and metastasis and is therefore a highly suitable target for the development of anti-cancer therapeutics. Previous efforts have concentrated on inhibiting the serine protease activity of uPA with small molecules. But this strategy neglects interactions of the uPAR/uPA complex with cell signaling proteins that are independent of the proteolytic activity of uPA. In this application, we follow a unique approach that seeks to target the uPAR/uPA protein-protein interaction using small molecules with the expectation that these molecules will exhibit the dual effect of blocking proteolysis and signaling. Our initial efforts have been highly rewarding. A preliminary structure-based computational screen has led to 8 active compounds. Cell culture-based studies reveal that a number of these compounds block MDA-MB-231 tumor cell adhesion, migration, and invasion. Compounds were also found to inhibit lung H1299 tumor cell proliferation. Biochemical analyses reveal that that these compounds also block integrin binding to the uPAR/uPA complex. Our objective in this grant application is to optimize the pharmacokinetic properties and potency of three of these compounds to block tumor invasion and metastasis in vivo. To that end, a multidisciplinary research program involving computational biology, synthetic chemistry, biophysical chemistry, structural biology, and cell biology will be followed. The first aim will consist of creating a virtual combinatorial library of compounds based on our lead molecules, ranking these compounds based on predicted potency, and predicting pharmacokinetic properties of the most potent compounds. The second aim will involve the chemical synthesis of the most promising compounds, followed by an assessment of their activity using a fluorescence polarization assay. The most potent compounds are further characterized with isothermal titration calorimetry. We then solve the three-dimensional structure of these compounds in complex with the urokinase receptor using x-ray diffraction. Finally, the third aim will consist of assessing the cellular efficacy of the most selective and potent inhibitors in MDA-MB-231 and other tumor cell lines, and to perform preliminary in vivo dosage studies to set the stage for future studies in mice xenografts.

项目总结/摘要尿激酶受体（uPAR）是一种GPI锚定蛋白，作为丝氨酸蛋白酶的受体尿激酶（uPA）。在对接到其受体后，uPA被激活，这是导致细胞周围蛋白水解的事件以及细胞外基质（肿瘤和脉管系统之间的主要屏障）的降解。但现在普遍认为，虽然uPAR不能信号传导，但uPAR/uPA复合物相互作用并激活许多细胞表面受体，如整联蛋白、受体酪氨酸激酶和GPCR。uPA/uPA 相互作用几乎涉及肿瘤侵袭和转移的每一步，因此是一种非常适合用于开发抗癌治疗剂的靶点。以前的努力集中在用小分子抑制uPA的丝氨酸蛋白酶活性。但这种策略忽略了 uPAR/uPA复合物与不依赖于uPA蛋白水解活性的细胞信号蛋白。在这应用中，我们遵循一种独特的方法，旨在靶向uPAR/uPA蛋白质-蛋白质相互作用，期望这些分子将表现出阻断蛋白水解的双重作用和信号。我们最初的努力是非常有益的。一个初步的基于结构的计算筛选得到8种活性化合物。基于细胞培养的研究表明，这些化合物中的一些阻断MDA-MB-231肿瘤细胞粘附、迁移和侵袭。还发现化合物抑制肺 H1299肿瘤细胞增殖。生化分析表明，这些化合物也阻断整合素，与uPAR/uPA复合物结合。我们申请资助的目的是优化药物的药代动力学这些化合物中的三种在体内阻断肿瘤侵袭和转移的性质和效力。与 end，一个多学科的研究计划，涉及计算生物学，合成化学，生物物理化学、结构生物学和细胞生物学将被遵循。第一个目标将包括创建一个虚拟的基于我们的先导分子的化合物组合库，根据预测效力，并预测最有效化合物的药代动力学性质。第二个目的将涉及最有前途的化合物的化学合成，然后评估其使用荧光偏振测定法测定活性。最有效的化合物进一步表征为：等温滴定量热法然后，我们解决了这些化合物的三维结构，与尿激酶受体的结合。最后，第三个目标将包括评估细胞在MDA-MB-231和其他肿瘤细胞系中最具选择性和有效的抑制剂的疗效，并进行初步的体内剂量研究，为今后在小鼠异种移植物中的研究奠定基础。

项目成果

期刊论文数量（11）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Small-molecule binding sites to explore protein-protein interactions in the cancer proteome.

DOI：
10.1039/c6mb00231e
发表时间：
2016-10-20
期刊：
Molecular bioSystems
影响因子：
0
作者：
Xu D;Jalal SI;Sledge GW;Meroueh SO
通讯作者：
Meroueh SO

Support vector regression scoring of receptor-ligand complexes for rank-ordering and virtual screening of chemical libraries.

DOI：
10.1021/ci200078f
发表时间：
2011-09-26
期刊：
Journal of chemical information and modeling
影响因子：
5.6
作者：
Li L;Wang B;Meroueh SO
通讯作者：
Meroueh SO

BioDrugScreen: a computational drug design resource for ranking molecules docked to the human proteome.

DOI：
10.1093/nar/gkp852
发表时间：
2010-01
期刊：
Nucleic acids research
影响因子：
14.9
作者：
Li L;Bum-Erdene K;Baenziger PH;Rosen JJ;Hemmert JR;Nellis JA;Pierce ME;Meroueh SO
通讯作者：
Meroueh SO

In silico docking and electrophysiological characterization of lacosamide binding sites on collapsin response mediator protein-2 identifies a pocket important in modulating sodium channel slow inactivation.

崩解蛋白反应介导蛋白 2 上拉科酰胺结合位点的计算机对接和电生理学表征确定了一个在调节钠通道缓慢失活中重要的口袋。

DOI：
10.1074/jbc.m110.128801
发表时间：
2010
期刊：
The Journal of biological chemistry
影响因子：
0
作者：
Wang,Yuying;Brittain,JoelM;Jarecki,BrianW;Park,KiDuk;Wilson,SarahM;Wang,Bo;Hale,Rachel;Meroueh,SamyO;Cummins,TheodoreR;Khanna,Rajesh
通讯作者：
Khanna,Rajesh

Effect of Binding Pose and Modeled Structures on SVMGen and GlideScore Enrichment of Chemical Libraries.