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：
9254523
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 2021-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9254523
关键词：
Adult Antibodies Antineoplastic Agents Apoptosis Back Binding Biological Biological Assay Biology Calorimetry Caspase Cell Line Cell Proliferation Complex Computer Assisted Crystallization Dimensions Drug Design Drug Kinetics 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 Molecular Models Multi-Drug Resistance Mus Nature Neoplasm Metastasis Nodal Normal Cell Normal tissue morphology Oncogenes Patients Peptides Pharmaceutical Chemistry Pharmaceutical Preparations Pharmacodynamics 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 therapy cancer type design drug development efficacy study feeding improved in vitro Assay in vivo inhibitor-of-apoptosis protein inhibitor/antagonist melanoma member molecular modeling molecular shape novel peptidomimetics 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（由申请人提供）：生存素的表达与癌症耐药性和患者生存率差呈正相关。生存素在大多数类型的癌症中普遍表达，但在正常组织中表达非常低，因此，开发新型选择性生存素抑制剂具有潜力。我们最近发现了一种由化合物 UC-112 提供的新支架，可强烈抑制癌细胞增殖，选择性降解其他 IAP 中的存活蛋白，并且基于这些初步研究，我们的总体假设是，UC-112 的新型支架对生存素具有出色的选择性。该项目的目标是利用集成结构生物学来优化 UC-112 支架。目标 1. 基于 UC-112 支架进行计算机辅助药物设计并迭代优化抗癌活性。根据 survivin-Smac 复合物的结构和 UC-112 支架的预测分子模型，我们将设计和合成 UC-112 类似物的重点组，以优化其活性并阐明结构-活性关系。新化合物使用一组人类黑色素瘤细胞系测定和三维集落形成测定来鉴定多达 20 种最佳 UC-112 类似物（标准：IC50 < 100 nM 并且可以克服多药耐药性），以推进目标 2。目标 2定义新开发的 UC-112 类似物的生物学作用机制目标 2.1：UC-112 类似物相互作用的结构表征。我们将使用 ITC 和 SPR 测量来确认存活蛋白与其他 IAP 的靶向抑制和选择性。我们还将解析存活蛋白与有效 UC-112 类似物复合物的 X 射线晶体结构。返回目标 1，优化分子模型以实现更有效的结构优化。目标 2.2：评估 UC-112 对存活蛋白稳定性和/或之间的阻断的影响。目标 2.3：研究 UC-112 类似物是否在具有不同生存素表达水平的癌细胞中诱导不同程度的细胞凋亡。目标 2.4：确定 survivin 和 caspase 的任何脱靶效应。使用 Pulldowns 和抗体阵列分析来确定新的类似物以及其他 IAP 或新 UC-112 类似物的潜在靶标的选择性。 3.确定选择性生存素抑制剂的体内抗癌活性。 3.1：评估化合物稳定性、药代动力学 (PK) 和药效学 (PD) 特性，选择最多 6 种最佳 UC-112 类似物进行后续体内功效研究目标 3.2：评估所选存活蛋白抑制剂对人类黑色素瘤肿瘤生长的抗癌活性。体内（小鼠）和对正常细胞/组织的潜在毒性目标 3.3：评估我们的选择性生存素抑制剂的治疗能力。使用我们建立的实验性肺转移模型进行体内黑色素瘤转移。