Restoration of Tumor Suppression Activity in Malignant Melanoma

描述（由申请人提供）：我们和其他人发现S100B不仅是一种临床标记物，而且它与p53结合，解离p53四聚体，并下调p53蛋白水平和功能。该项目的目标是抑制S100B-p53相互作用，恢复该癌症中wt p53蛋白水平。计算机辅助药物设计（CADD）、核磁共振、热力学结合和p53功能分析将用于发现与Ca2+负载的S100B上p53位点结合并抑制S100B-p53相互作用的先导化合物。通过核磁共振方法和对接协议的SAR（结构/活性关系）也将用于寻找结合Ca2+S100B上其他位点的化合物，这些位点靠近p53结合位点。s100b药物配合物的三维表征将使用核磁共振波谱和x射线晶体学进行。这将包括新发现的配合物和几种现有的s100b药物配合物(S100BKD<10 ？M)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。有机合成将以三维结构数据和CADD导联优化方法为指导。这些类似物的测试将使用核磁共振，竞争结合研究和生物。作为一个新的目标，这是本次竞争性修订的主题，将在黑色素瘤小鼠模型中进行有前途的化合物抑制/消除肿瘤的能力的体内测试。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，首先帮助我们将设计/合成工作的重点放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优先使用哪些化合物是绝对必要的。我们将雇用两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的几家工厂的众多员工提供工作。

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.