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: 7424998
• 负责人: David Joseph Weber
• 金额: $ 48.71万
• 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 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7424998
• 关键词: Actins; Affinity; Animal Model; Antibodies; Antisense RNA; Apoptosis; Astrocytoma; Binding; Binding Sites; Biological Assay; CDKN1A gene; Chemicals; Complex; Computer Assisted; Data; Dose; Drug Design; Figs - dietary; Future; Goals; Human; In Vitro; Lead; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of kidney; Mature T-Lymphocyte; Melanoma Cell; Modification; Molecular Weight; NMR Spectroscopy; Organic Synthesis; Patients; Peptides; Pharmaceutical Preparations; Protein Binding; Protein p53; Proteins; Protocols documentation; Research Personnel; Site; Small Interfering RNA; Solutions; Structure; Structure-Activity Relationship; TP53 gene; Testing; Therapeutic; Thermodynamics; Tumor Suppression; X ray spectroscopy; X-Ray Crystallography; analog; design; inhibitor/antagonist; leukemia; melanoma; mouse model; neurotrophic protein S100beta; oncoprotein p21; programs; restoration; three dimensional structure; tumor

DESCRIPTION (provided by applicant): Our lab has shown that the S100B protein binds to wild-type p53 in primary malignant melanoma, dissociates the p53 tetramer, and down-regulates p53-dependent tumor suppression; therefore, it is important to develop inhibitors of the S100B-p53 interaction to restore wild-type p53 activity in this cancer. As a proof of principle, we have also demonstrated that inhibiting S100B with small interfering antisense RNA (siRNASIOOB) restores wild-type p53 tumor suppressor activity in primary malignant melanoma. Accordingly, we hypothesize that low molecular weight compounds can be rationally designed to bind the well-defined p53 binding site on S100B with high affinity and inhibit the S100B-p53 interaction. This hypothesis will be tested further in the proposed study, via discovery and synthesis of such molecules with the following specific aims. In Aim 1, computer aided drug design (CADD), combined with high- throughput/automated NMR, thermodynamic binding and p53 functional assays will be used to discover lead compounds that bind S100B and inhibit the S100B-p53 interaction. In Aim 2, 3D structures of S100B-drug complexes will be determined using NMR spectroscopy and/or X-ray crystallography. Such structure determinations are already underway. In Aim 3, optimization of lead compounds that inhibit the S100B-p53 interaction will be performed via chemical modifications. Organic syntheses will be guided by 3D structural data (from Aim 2) and CADD lead optimization approaches. Testing of new analogues will be performed using existing thermodynamic binding and biological assays (as in Aim 1). With this strategy, it is our aim to discover/synthesize new compounds that bind S100B and restore p53 activity in malignant melanoma. In the future, the most promising compounds will be examined for efficacy in treating melanoma in animal models. Inhibitors such as these will likely have therapeutic value for treatment of other cancers that have elevated S100B levels and wild-type p53 such as astrocytomas, renal tumors and malignant mature T-cells in leukemia patients.

描述（由申请人提供）：我们的实验室表明，S100B蛋白在原发性恶性黑色素瘤中与野生型p53结合，分离p53四聚体，并下调依赖p53依赖性肿瘤。因此，重要的是开发S100B-P53相互作用的抑制剂，以恢复该癌症中野生型p53活性。作为原则的证明，我们还证明，用小的干扰反义RNA（siRNASIOOB）抑制S100B可恢复原发性恶性黑色素瘤中野生型p53肿瘤抑制剂的活性。因此，我们假设低分子量化合物可以合理设计，以结合具有高亲和力的S100B上定义明确的p53结合位点并抑制S100B-P53相互作用。该假设将在拟议的研究中通过以下特定目的发现和合成此类分子进行进一步检验。在AIM 1中，计算机辅助药物设计（CADD）与高吞吐量/自动化NMR相结合，热力学结合和p53功能分析将用于发现结合S100B并抑制S100B-P53相互作用的铅化合物。在AIM 2中，将使用NMR光谱和/或X射线晶体学确定S100B-prug配合物的3D结构。这种结构确定已经在进行中。在AIM 3中，将通过化学修饰进行抑制S100B-P53相互作用的铅化合物的优化。有机合成将以3D结构数据（来自AIM 2）和CADD铅优化方法指导。将使用现有的热力学结合和生物学测定（如AIM 1）进行新类似物的测试。通过这种策略，我们的目标是发现/合成结合S100B并恢复恶性黑色素瘤活性的新化合物。将来，将检查最有希望的化合物在动物模型中治疗黑色素瘤的功效。诸如此类的抑制剂可能具有治疗其他S100B水平升高和野生型p53的癌症的治疗价值，例如星形胶质细胞瘤，肾肿瘤和白血病患者的恶性成熟T细胞。