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: 8188169
• 负责人: David Joseph Weber
• 金额: $ 48.08万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8188169
• 关键词: Address; Advisory Committees; Affinity; Animal Model; Antisense RNA; Apoptosis; Astrocytoma; Binding; Binding Sites; Biological Assay; Biology; Biopsy; Cellular Assay; Chemicals; Clinical; Clinical Markers; Clinical Trials; Complex; Computer Assisted; Data; Databases; Disease; Disease Progression; Drug Design; Engineering; Excretory function; Exhibits; Funding; Future; Goals; Grant; Growth Factor; Human; Immunoprecipitation; In Vitro; Kidney; Kidney Neoplasms; Knowledge; Lead; Legal patent; Ligands; Malignant - descriptor; Malignant Epithelial Cell; Malignant Neoplasms; Maryland; Metabolism; Methods; Modification; Molecular Weight; Mutate; NMR Spectroscopy; Paper; Patients; Pharmaceutical Chemistry; Pharmaceutical Preparations; Phase II Clinical Trials; Poison; Polyubiquitination; Production; Property; Protein p53; Proteins; Public Health; Publications; Recurrence; Roentgen Rays; Site; Specificity; Staining method; Stains; Structure; Surface; Testing; Therapeutic; Thermodynamics; Time; Transcriptional Activation; Tumor Markers; Tumor Suppression; Tumor Suppressor Proteins; Ubiquitination; Universities; Woman; X-Ray Crystallography; absorption; analog; base; cancer cell; cell growth; design; efficacy evaluation; high throughput screening; improved; in vivo; inhibitor/antagonist; leukemia; melanoma; men; mouse model; neurotrophic protein S100beta; novel therapeutics; process optimization; prognostic indicator; restoration; three dimensional structure; tumor; tumor growth; tumor progression

DESCRIPTION (provided by applicant): In malignant melanoma (MM), we have shown that elevated levels of the tumor marker, S100B, binds directly to wild-type (wt) p53, dissociates the p53 tetramer, enhances hdm2-dependent ubiquitination of p53, and down-regulates p53-dependent tumor suppression functions; therefore, it is important to develop S100B inhibitors (SBiXs; X=compound number) to restore active p53 in this deadly cancer. As a proof of principle for a drug design project, inhibiting S100B with small interfering antisense RNA (siRNAS100B) and several SBiXs was shown to restore wild-type p53 levels and its downstream gene products, as necessary to induce cell growth arrest and apoptosis in malignant melanoma. We hypothesize that low molecular weight compounds can be designed to bind the well-defined p53 binding site on Ca2+ loaded S100B with higher affinity and specifically inhibit the Ca2+dependent S100B-p53 interaction to mimic the siRNAS100B effects. In the last granting period, thirty-eight publications, ten patent applications, and twenty protein data base submissions describe our progress at addressing these goals/hypotheses. Animal model studies with several SBiXs and a human clinical trial with SBi1 are also ongoing as a result of our progress. However, it is important that we engineer and/or synthesis new and improved SBiXs with higher affinity and better specificity towards inhibiting S100B. This will be achieved with the following specific aims: In Aim 1, computer aided drug design (CADD) combined with high-throughput screening methods (i.e. NMR, binding, and cellular assays) will be used to discover/design new compounds that bind S100B and inhibit the S100B-p53 interaction at lower concentrations and with higher specificity than those already discovered. Optimization of promising leads will be performed via chemical modifications as guided by 3D structural and computer aided drug design (CADD) lead optimization approaches. In Aim 2, 3D structures of S100B-SBiX complexes will be determined using NMR spectroscopy and/or X-ray crystallography to further characterize the binding surface on Ca2+S100B, so improved SBiXs can be designed and synthesized. Testing new analogues will be performed using existing thermodynamic binding and biological assays (as in Aim 1) with the most promising compounds examined for their ability to suppress/eliminate tumor growth in melanoma mouse models (in Aim 3). The in vivo data will be important for focusing our design/synthesis efforts on lead compounds or classes of compounds that have efficacy in vivo, and they will be used to set priorities for additional human clinical trials to be done in the future. It is our goal to discover/synthesize and/or to improve existing SBiXs to optimally restore p53 activity in human malignant melanoma. SBiXs may also have therapeutic value for treating other cancers with elevated S100B and wt p53 such as astrocytomas, renal tumors, and some forms of leukemia, so this will be explored in future endeavors. PUBLIC HEALTH RELEVANCE: The ongoing project (CA107331) addresses important public health concerns including understanding how the well-established tumor marker, S100B, contributes to cancer progression by down-regulating the tumor suppressor protein p53. This information impacts how we develop drugs to treat several cancers, including malignant melanoma, which has elevated levels of the S100B growth factor. Such molecules will be tested both in vitro and in vivo as necessary to initiate human clinical trials, as was done in the past with one S100B inhibitor, SBi1.

描述（由申请人提供）：在恶性黑素瘤（MM）中，我们已经表明肿瘤标志物S100 B的水平升高，直接结合野生型（wt）p53，解离p53四聚体，增强hdm 2依赖性p53泛素化，并下调p53依赖性肿瘤抑制功能;因此，开发S100 B抑制剂是重要的（SBiXs; X=化合物编号）来恢复这种致命癌症中的活性p53。作为药物设计项目的原理证明，用小干扰反义RNA（siRNAS 100 B）和几种SBiX抑制S100 B显示出恢复野生型p53水平及其下游基因产物，这是诱导恶性黑素瘤中细胞生长停滞和凋亡所必需的。我们假设，低分子量化合物可以被设计为以更高的亲和力结合负载Ca 2+的S100 B上明确定义的p53结合位点，并特异性抑制Ca 2+依赖性S100 B-p53相互作用以模拟siRNAS 100 B效应。在上一个授权期，38篇出版物，10项专利申请和20项蛋白质数据库提交描述了我们在解决这些目标/假设方面的进展。由于我们的进展，使用几种SBiX的动物模型研究和使用SBi 1的人体临床试验也在进行中。然而，重要的是我们工程化和/或合成新的和改进的SBiX，其对抑制S100 B具有更高的亲和力和更好的特异性。这将通过以下具体目标实现：在目标1中，计算机辅助药物设计（CADD）结合高通量筛选方法（即NMR，结合和细胞测定）将用于发现/设计新化合物，其结合S100 B并在较低浓度下抑制S100 B-p53相互作用，并且具有比已经发现的更高的特异性。将通过化学修饰进行有前途的引线的优化，以3D结构和计算机辅助药物设计（CADD）引线优化方法为指导。在目标2中，将使用NMR光谱和/或X射线晶体学来确定S100 B-SBiX复合物的3D结构，以进一步表征Ca 2 + S100 B上的结合表面，从而可以设计和合成改进的SBiX。将使用现有的热力学结合和生物学试验（如目标1所示）检测新的类似物，并检查最有希望的化合物在黑色素瘤小鼠模型中抑制/消除肿瘤生长的能力（如目标3所示）。体内数据对于将我们的设计/合成工作集中在具有体内功效的先导化合物或化合物类别上将是重要的，并且它们将用于设定未来要进行的其他人体临床试验的优先级。我们的目标是发现/合成和/或改进现有的SBiX以最佳地恢复人恶性黑素瘤中的p53活性。SBiX也可能对治疗S100 B和wt p53升高的其他癌症具有治疗价值，例如星形细胞瘤，肾肿瘤和某些形式的白血病，因此这将在未来的努力中进行探索。 公共卫生关系：正在进行的项目（CA 107331）解决了重要的公共卫生问题，包括了解公认的肿瘤标志物S100 B如何通过下调肿瘤抑制蛋白p53促进癌症进展。这一信息影响了我们如何开发治疗多种癌症的药物，包括恶性黑色素瘤，它具有升高的S100 B生长因子水平。这些分子将在体外和体内进行测试，以启动人类临床试验，就像过去对S100 B抑制剂SBi 1所做的那样。