Selective Targeting Survivin for Cancer Therapy

选择性靶向生存素用于癌症治疗

基本信息

批准号：
9922228
负责人：
WEI LI
金额：
$ 38.01万
依托单位：
UNIVERSITY OF TENNESSEE HEALTH SCI CTR
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-05-01 至 2022-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9922228
关键词：
3-Dimensional Adult Antibodies Antineoplastic Agents Apoptosis Back Binding Biological Biological Assay Calorimetry Caspase Cell Line Cell Proliferation Complex Computer Assisted Crystallization Drug Design Drug Targeting Drug resistance Drug toxicity Evaluation Failure Fluorescence Polarization Human Investigation Libraries Measurement Mediating Melanoma Cell Metastatic Neoplasm to the Lung Mitochondria Modeling Molecular Molecular Biology Multi-Drug Resistance Mus Nature Neoplasm Metastasis Nodal Normal Cell Normal tissue morphology Oncogenes Patients Peptides Pharmaceutical Chemistry Pharmaceutical Preparations Physiologic pulse Property Protein Binding Domain Proteins Reporting Roentgen Rays Shapes Structure Structure-Activity Relationship Surface Plasmon Resonance Testing Tissues Titrations Toxic effect Ubiquitination Western Blotting Work analog anticancer activity base cancer cell cancer drug resistance cancer therapy cancer type design drug development efficacy study improved in vitro Assay in vivo inhibitor-of-apoptosis protein inhibitor/antagonist melanoma member molecular modeling molecular shape novel peptidomimetics pharmacokinetics and pharmacodynamics protein protein interaction public health relevance scaffold screening structural biology survivin tumor growth tumor progression

项目摘要

DESCRIPTION (provided by applicant): The expression of survivin positively correlates with cancer drug resistance and poor patient survival. Survivin is ubiquitously expressed in most types of cancer, but has very low expression in normal tissues. Thus, developing novel selective survivin inhibitors as potential cancer therapies is highly significant. We recently discovered a new scaffold provided by compound UC-112. UC-112 strongly inhibits cancer cell proliferation, selectively degrades survivin among other IAPs, and potently suppresses melanoma tumor growth in vivo. Based on these preliminary studies, our overall hypothesis is that the novel scaffold of UC-112 is exquisitely selective for survivin. The objective of this project is to optimize the UC-112 scaffold using integrated structural biology, molecular modeling, medicinal chemistry, and molecular biology approaches. Aim 1. Perform computer-aided drug design based on the UC-112 scaffold and iteratively optimize the anticancer activity. Aim 1.1: Using the crystal structure of survivin-Smac complex and our predictive molecular models for the UC-112 scaffold, we will design and synthesize focused sets of UC-112 analogs to optimize their activity and elucidate the structure-activity relationships. Aim 1.2: Screen the new compounds using a panel of human melanoma cell lines and three-dimensional colony formation assays to identify up to 20 best UC-112 analogs (criteria: IC50 < 100 nM and can overcome multidrug resistance) to be advanced to Aim 2. Aim 2. Define the biological mechanism(s) of action of newly developed UC-112 analogs. Aim 2.1: Structural characterization of UC-112 analogs interacting with survivin and other IAPs. We will confirm the on-target survivin inhibition and selectivity among other IAPs using ITC and SPR measurements. We will also solve the X-ray crystal structures of survivin in complex with potent UC-112 analogs. This information will feed back to Aim 1 to optimize the molecular models for more efficient structural optimization. Aim 2.2: Evaluation of the effect of UC-112 on survivin stability and/or blockage between survivin and caspases using ubiquitination, pulse-chase, and Western blot analyses. Aim 2.3: Investigation of whether UC-112 analogs induce differentiated degrees of apoptosis in cancer cells with distinct survivin expression levels. Aim 2.4: Determination of any off-target effects of the new analogs and selectivity among other IAPs or additionally potential targets of the new UC-112 analogs using pulldowns and antibody array analysis. Aim 3. Determine the anticancer activity of selective survivin inhibitors in vivo. Aim 3.1: Evaluate compound stability, pharmacokinetics (PK), and pharmacodynamics (PD) properties to select up to six best UC-112 analogs for subsequent in vivo efficacy studies. Aim 3.2: Evaluate anticancer activities of the selected survivin inhibitors against human melanoma tumor growth in vivo (in mice) and the potential toxicity to normal cells/tissues. Aim 3.3: Evaluate the ability of our selective survivin inhibitors to treat melanoma metastasis in vivo usin our established experimental lung metastasis model.

描述（由适用提供）：生存的表达与癌症耐药性和患者生存不良相关。 Survivin在大多数类型的癌症中无处不在，但在正常组织中的表达非常低。这是将新型的选择性生存抑制剂作为潜在的癌症疗法非常重要的。我们最近发现了Compound UC-112提供的新脚手架。 UC-112强烈抑制癌细胞的增殖，在其他IAP中有选择性降解，并可能抑制体内黑色素瘤肿瘤的生长。基于这些初步研究，我们的总体假设是，UC-112的新型支架对于属于overivin都是选择性的。该项目的目的是使用综合结构生物学，分子建模，药物化学和分子生物学方法来优化UC-112支架。 AIM 1。根据UC-112支架进行计算机辅助药物设计，并迭代地优化抗癌活性。 AIM 1.1：使用Survivin-SMAC复合物的晶体结构以及我们针对UC-112支架的预测分子模型，我们将设计和合成UC-112类似物的集中集合集，以优化其活性并阐明结构 - 活动关系。目标1.2：屏幕新化合物使用一组人黑色素瘤细胞系和三维菌落形成测定法，以识别多达20种最佳UC-112类似物（标准：IC50 <100 nm，可以克服多种耐药性），以提高到目标2。AIM2。定义生物学机制的新型生物学机制。 AIM 2.1：UC-112类似物与Survivin和其他IAP相互作用的结构表征。我们将使用ITC和SPR测量值确认其他IAP的靶标的Survivin抑制和选择性。我们还将与潜在的UC-112类似物中的复合物中求解Survivin的X射线晶体结构。该信息将反馈到AIM 1，以优化分子模型，以进行更有效的结构优化。 AIM 2.2：使用泛素化，脉冲练习和Western印迹分析评估UC-112对Survivin和caspase之间的Survivin稳定性和/或阻塞的影响。 AIM 2.3：研究UC-112是否诱导具有不同生存表达水平的癌细胞中分化的凋亡程度。 AIM 2.4：使用下拉和抗体阵列分析确定其他IAP的新类似物和选择性的任何脱靶效应和新UC-112类似物的其他潜在目标。 AIM 3。确定体内选择性Survivin抑制剂的抗癌活性。目的 3.1：评估复合稳定性，药代动力学（PK）和药效学（PD）特性，以选择多达六种最佳UC-1112类似物，以进行随后的体内效率研究。 AIM 3.2：评估选定的Survivin抑制剂对体内黑色素瘤肿瘤生长的抗癌活性（在小鼠中）以及对正常细胞/组织的潜在毒性。 AIM 3.3：评估我们选择性遗传抑制剂在体内治疗黑色素瘤转移的能力，我们已建立的实验性肺转移模型。