Restoration of Tumor Suppression Activity in Malignant Melanoma

恶性黑色素瘤肿瘤抑制活性的恢复

基本信息

批准号：
7812305
负责人：
David Joseph Weber
金额：
$ 37.63万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
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：
2009-09-17 至 2011-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7812305
关键词：
Address Affinity Animal Model Astrocytoma Baltimore Binding Binding Sites Biological Biological Assay Cellular Assay Chemicals Clinical Markers Clinical Trials Collaborations Complex Computer Assisted Data Drug Design Employee Engineering Future Goals Grant Human In Vitro Lead Malignant Neoplasms Maryland Melanoma Cell Methods Modification Molecular Weight NMR Spectroscopy Organic Synthesis Peptides Pharmaceutical Preparations Protein p53 Protocols documentation Public Health Renal Cell Carcinoma Scheme Schools Site Solutions Specificity Structure Structure-Activity Relationship TP53 gene Testing Texas Therapeutic Thermodynamics Time Tumor Suppression Universities Veterinary Medicine Work X-Ray Crystallography analog design in vivo inhibitor/antagonist leukemia melanoma mouse model protein protein interaction public health relevance restoration tumor

项目摘要

DESCRIPTION (provided by applicant): We and others have discovered that S100B is not just a clinical marker, but that it binds to p53, dissociates the p53 tetramer, and down-regulates p53 protein levels and function in malignant melanoma. The goal of this project is to inhibit the S100B-p53 interaction and restore wt p53 protein levels in this cancer. Computer aided drug design (CADD), NMR, thermodynamic binding, and p53 functional assays will be used to discover lead compounds that bind to the p53 site on Ca2+loaded S100B and inhibit the S100B-p53 interaction. The SAR (structure/activity relationship) by NMR approach and DOCKING protocols will also be used to find compounds that bind other site(s) on Ca2+S100B, which are proximal to the p53 binding site. 3D characterization of S100B-drug complexes will be performed using NMR spectroscopy and X-ray crystallography. This will include structure determinations of new complexes discovered and several existing S100B-drug complexes (S100BKD<10 ?M) that inhibit the S100B-p53 interaction in vitro and in primary malignant melanoma cells. Compounds that inhibit the S100B-p53 interaction will be optimized to bind S100B more tightly via chemical modifications. Organic syntheses will be guided by 3D structural data and CADD lead optimization approaches. Testing of such analogues will be done using NMR, competition binding studies, and biological. As a new Aim, which is the topic of this competitive revision, will be to do in vivo testing of promising compound for their ability to suppress/eliminate tumors in melanoma mouse models. Such work will be done in collaboration with Dr. Danna Zimmer at the Texas A&M School of Veterinary Medicine. These in vivo data will be critically important first by helping us focus our design/synthesis efforts only on lead compounds or classes of compounds that have efficacy in vivo. Additionally, these data are absolutely necessary to set priorities for which compounds to pursue in human clinical trials. We will hire 2 new technicians to complete these studies as well as provide work for numerous employees at several facilities at both the University of Maryland and at Texas A & M. PUBLIC HEALTH RELEVANCE: The ongoing project (CA197331) as well as the new work proposed in this competitive revision application addresses many important public health concerns including providing drugs to treat several cancers including malignant melanoma. This work also provides general principles for inhibiting protein-protein interactions, which is at the forefront of drug-design challenges.

描述（由申请人提供）：我们和其他人发现 S100B 不仅是一种临床标志物，而且它与 p53 结合，解离 p53 四聚体，并下调恶性黑色素瘤中的 p53 蛋白水平和功能。该项目的目标是抑制 S100B-p53 相互作用并恢复该癌症中 wt p53 蛋白水平。计算机辅助药物设计 (CADD)、NMR、热力学结合和 p53 功能测定将用于发现与 Ca2+ 负载的 S100B 上的 p53 位点结合并抑制 S100B-p53 相互作用的先导化合物。 NMR 方法和 DOCKING 方案的 SAR（结构/活性关系）也将用于寻找与 Ca2+S100B 上其他位点（靠近 p53 结合位点）结合的化合物。 S100B-药物复合物的 3D 表征将使用 NMR 光谱和 X 射线晶体学进行。这将包括发现的新复合物和几种现有的 S100B-药物复合物 (S100BKD<10 μM) 的结构测定，这些复合物在体外和原发性恶性黑色素瘤细胞中抑制 S100B-p53 相互作用。抑制 S100B-p53 相互作用的化合物将被优化，通过化学修饰更紧密地结合 S100B。有机合成将以 3D 结构数据和 CADD 先导优化方法为指导。此类类似物的测试将使用核磁共振、竞争结合研究和生物学来完成。本次竞争性修订的一个新目标是对有前景的化合物在黑色素瘤小鼠模型中抑制/消除肿瘤的能力进行体内测试。此类工作将与德克萨斯农工大学兽医学院的 Danna Zimmer 博士合作完成。这些体内数据首先将非常重要，因为它们可以帮助我们将设计/合成工作仅集中在先导化合物或在体内有效的化合物类别上。此外，这些数据对于确定在人体临床试验中追求的化合物的优先顺序是绝对必要的。我们将聘请 2 名新的技术人员来完成这些研究，并为马里兰大学和德克萨斯农工大学多个机构的众多员工提供工作。公共健康相关性：正在进行的项目 (CA197331) 以及本竞争性修订申请中提出的新工作解决了许多重要的公共健康问题，包括提供治疗包括恶性黑色素瘤在内的多种癌症的药物。这项工作还提供了抑制蛋白质-蛋白质相互作用的一般原则，这是药物设计挑战的最前沿。