Restoration of Tumor Suppression Activity in Malignant Melanoma

恶性黑色素瘤肿瘤抑制活性的恢复

• 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
• 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

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S100 B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S100 B-p53相互作用并恢复这种癌症中的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结合到负载Ca 2+的S100 B上的p53位点并抑制S100 B-p53相互作用的先导化合物。通过NMR方法和对接方案的SAR（结构/活性关系）也将用于发现结合Ca 2 + S100 B上的其它位点的化合物，所述其它位点邻近p53结合位点。S100 B-药物复合物的3D表征将使用NMR光谱学和X射线晶体学进行。这将包括新发现的复合物和几个现有的S100 B-药物复合物（S100 BKD <10？M），其在体外和在原发性恶性黑素瘤细胞中抑制S100 B-p53相互作用。抑制S100 B-p53相互作用的化合物将被优化，以通过化学修饰更紧密地结合S100 B。有机合成将由3D结构数据和CADD铅优化方法指导。此类类似物的测试将使用NMR、竞争结合研究和生物学进行。作为一个新的目标，这是这个竞争性修订的主题，将在体内测试有前途的化合物在黑色素瘤小鼠模型中抑制/消除肿瘤的能力。这项工作将与德克萨斯A&M兽医学院的Danna Zimmer博士合作完成。这些体内数据将是至关重要的，首先帮助我们把我们的设计/合成工作只集中在先导化合物或化合物的类，有疗效的体内。此外，这些数据对于确定在人体临床试验中追求哪些化合物的优先级是绝对必要的。我们将聘请2名新技术人员来完成这些研究，并为马里兰州大学和德克萨斯A & M大学的多个设施的众多员工提供工作。 公共卫生相关性：正在进行的项目（CA 197331）以及在此竞争性修订申请中提出的新工作解决了许多重要的公共卫生问题，包括提供治疗多种癌症（包括恶性黑色素瘤）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。