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）：四种癌基因 c-MYC、bcl-2、VEGF 和 HIF-1a 在其启动子区域均含有对转录激活至关重要的 polyG/polyC 区域。 DNA G-四链体二级结构的出现已在这四种癌基因的启动子区域中得到证实，并且已被证明是一种转录调节剂。虽然DNA G-四链体是有希望的新药物靶点，但对其作为癌症治疗靶点的潜力的评估取决于对生物学相关的G-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 启动子区域中 G 四链体的主要形式，它们似乎代表了 VEGF/HIF 的三种不同的基本 G 四链体结构-1a G-四联体是相同的基本类型。启动子 G-四链体的不同分子结构使这些结构成为途径特异性药物设计的有吸引力的目标。在本提案中，我们打算定义每个 G-四联体及其药物复合物的具体分子结构。获得的结构信息将与生物数据相关联，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合理药物设计提供重要基础。我们将测试我们的假设，即每个启动子 G-四联体可以被不同的小分子药物化合物特异性靶向。除了可以通过与不同的G-四链体核心结构的相互作用来实现选择性的原理之外，我们期望还可以通过外部环和产生结合袋的封端结构内的相互作用来实现选择性。在这方面，原理证明很重要。我们将结合使用 NMR、CD、分子建模和微量热数据以及适当的突变启动子元件。我们的主要方法，高场核磁共振波谱，代表了生物学相关的 G-四链体结构测定的主要工具，因为这种结构的结晶困难。我们的最终目标是使用基于结构的方法合理设计小分子 G-四链体相互作用化合物，专门针对每个启动子特有的 G-四链体结构并调节基因转录。具体来说，我们计划1）确定c-MYC启动子区域形成的生物学相关G-四链体的分子结构及其药物相互作用； 2) 确定bcl-2启动子区形成的生物学相关G-四链体的分子结构及其药物相互作用； 3) 确定 VEGF/HIF-1a 启动子区域形成的生物学相关 G-四链体的分子结构及其药物相互作用。