G-quadruplexes formed in human oncogene promoters and their drug complexes

人类癌基因启动子及其药物复合物中形成的G-四链体

基本信息

批准号：
7659417
负责人：
DANZHOU YANG
金额：
$ 28.47万
依托单位：
UNIVERSITY OF ARIZONA
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-03 至 2012-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7659417
关键词：
Adopted Affinity Back Binding Biological Biological Assay Chemicals Complex Crystallization Data Drug Compounding Drug Delivery Systems Drug Design Drug Interactions Elements Evaluation Fluorescence Resonance Energy Transfer G-Quartets Gene Structure Genes Genetic Transcription Human Hybrids Lateral Libraries MYC gene Malignant Neoplasms Modeling Molecular Conformation Molecular Models Molecular Structure NMR Spectroscopy Oncogenes Pathway interactions Pattern Pharmaceutical Preparations Phase I Clinical Trials Promoter Regions Property Research Personnel Screening procedure Side Specificity Structure Testing Thermodynamics Transcriptional Activation Transcriptional Silencer Elements Vascular Endothelial Growth Factors Work Yang base design drug development improved insight microcalorimetry molecular modeling mutant novel programs promoter small molecule therapeutic target tool

项目摘要

DESCRIPTION (provided by applicant): The four oncogenes, c-MYC, bcl-2, VEGF, and HIF-1a, all contain polyG/polyC tracts in their promoter regions critical for transcriptional activation. The occurrence of DNA G-quadruplex secondary structures has been demonstrated in the promoter regions of these four oncogenes and has been shown to be a transcriptional modulator. While the DNA G-quadruplexes are promising new drug targets, the evaluation of their potential as cancer therapeutic targets depends on the understanding of biologically relevant G-quadruplex structures. Our preliminary studies have allowed us to identify the predominant forms of G-quadruplexes in the promoter regions of c-MYC, bcl-2, VEGF, and HIF-1a, which appear to represent three different basic G-quadruplex structures with VEGF/HIF-1a G-quadruplexes being the same basic type. The different molecular structures of the promoter G-quadruplexes make these structures attractive targets for pathway-specific drug design. In this proposal we intend to define the specific molecular structure of each G-quadruplex and its drug-complex(es). The structural information obtained will be correlated with the biological data to understand the effective gene modulation. Insight into the structures of the promoter G-quadruplexes and their drug complexes will provide an important basis for structure-based rational drug design. We will test our hypothesis that each promoter G-quadruplex can be specifically targeted by different small molecule drug compounds. In addition to the principle that selectivity can be achieved by interactions with different G-quadruplex core structures, we expect that selectivity can also be achieved by interactions within the external loops and capping structures in which binding pockets are generated. Proof of principle will be important in this regard. We will use a combination of NMR, CD, molecular modeling, and microcalorimetry data in concert with appropriate mutant promoter elements. Our primary approach, high field NMR spectroscopy, represents a major tool for structure determination of biologically relevant G-quadruplexes, due to the difficulty of crystallization of such structures. Our ultimate objective is to use a structure-based approach to rationally design small molecule G-quadruplex-interactive compounds that specifically target the G-quadruplex structure unique to each promoter and modulate gene transcription. Specifically, we plan to 1) determine the molecular structure of the biologically relevant G-quadruplex formed in the promoter region of c-MYC and its drug interactions; 2) determine the molecular structure of the biologically relevant G-quadruplex formed in the promoter region of bcl-2 and its drug interactions; and 3) determine the molecular structure of the biologically relevant G-quadruplex formed in the promoter region of VEGF/HIF-1a and its drug interactions.

描述（由申请人提供）：四个Oncogenes，C-Myc，Bcl-2，VEGF和HIF-1A，均包含其启动子区域中的多头性/polyc Tracts，这对于转录激活至关重要。在这四种癌基因的启动子区域已证明了DNA G四链体二级结构的发生，并已被证明是转录调节剂。尽管DNA G四链体是有希望的新药物靶标，但作为癌症治疗靶标的潜力的评估取决于对生物学相关的G Qu-四链体结构的理解。我们的初步研究使我们能够确定C-Myc，Bcl-2，VEGF和HIF-1A的启动子区域中G-四链体的主要形式，它们似乎代表了三种不同的基本G Qu-四链体结构，具有VEGF/HIF-1A G-1A G-g-Quadruplexes是相同的基本类型。启动子G四链体的不同分子结构使这些结构具有特异性药物设计的吸引力。在此提案中，我们打算定义每个G四链体的特定分子结构及其药物复合物（ES）。获得的结构信息将与生物学数据相关，以了解有效的基因调节。深入了解启动子G-四链体及其药物复合物的结构将为基于结构的理性药物设计提供重要的基础。我们将测试我们的假设，即每个启动子G-四链体可以由不同的小分子药物化合物特别针对。除了通过与不同G Quadruplex核心结构的相互作用来实现选择性的原则外，我们希望通过在外部环中的相互作用和生成结合口袋的外部循环和上限结构中的相互作用来实现选择性。原则证明在这方面将很重要。我们将与适当的突变启动子元素一起使用NMR，CD，分子建模和微量钙化数据的组合。由于这种结构的结晶难度，我们的主要方法是高场NMR光谱法代表了生物学相关的G四链体的结构确定的主要工具。我们的最终目标是使用基于结构的方法来合理设计小分子G-四链体相互作用化合物，该化合物专门针对每个启动子独有的G四链体结构并调节基因转录。具体而言，我们计划1）确定在C-Myc启动子区域形成的生物学相关的G季链体的分子结构及其药物相互作用； 2）确定在Bcl-2启动子区域及其药物相互作用中形成的生物学相关的G四链体的分子结构； 3）确定在VEGF/HIF-1A启动子区域形成的生物学相关的G四链体的分子结构及其药物相互作用。