Discovery of small molecule inhibitors of c-Myc/Mac dimerization and DNA binding

发现 c-Myc/Mac 二聚化和 DNA 结合的小分子抑制剂

基本信息

批准号：
8332272
负责人：
RICHARD YOUNG
金额：
$ 4.88万
依托单位：
WHITEHEAD INSTITUTE FOR BIOMEDICAL RES
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-13 至 2013-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8332272
关键词：
American Animal Model Area Binding Binding Sites Biochemical Biological Biological Assay Biology Breast Burkitt Lymphoma Cancer cell line Cell Count Cell Line Cell Proliferation Cell Survival Cells Cellular Assay Chemicals Clinical Complex DNA DNA Binding Data Detection Development Diagnosis Dimerization Disease Dose Elements Exhibits Flow Cytometry Fluorescence Resonance Energy Transfer Gel Gene Expression Gene Targeting Genes Genetic Transcription Health Helix-Turn-Helix Motifs Heterodimerization Histocompatibility Testing Human In Vitro Lead Lung Malignant Neoplasms Measurement Metabolic Molecular Bank Monitor Morphologic artifacts Oncogenes Oncogenic Outcome Patients Pharmaceutical Chemistry Production Prostate Proteins Proxy Publishing Relapse Relative (related person)Reporter Genes Reporting Research Signal Transduction Technology Testing Therapeutic Transcript Transcription Factor Oncogene Transgenic Mice Translating Two-Hybrid System Techniques Validation Work Yeasts c-myc Genes cancer cell cell type effective therapy high throughput screening improved in vitro Assay in vivo inhibitor/antagonist insight meetings mouse model new technology novel novel therapeutics overexpression programs research study response small molecule therapeutic target transcription factor treatment strategy tumor tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): MYC is the most frequently amplified oncogene in human cancers occurring in a wide range of tissue types including breast, lung, and prostate. MYC overexpression occurs in at least 30% of all human cancers and frequently correlates with poor clinical outcome and increased chance of relapse. An estimated 450,000 Americans are diagnosed with a MYC-dependent cancer each year. These patients are in need of novel and effective treatment strategies. c-Myc is a helix-loop-helix transcription factor that drives a proliferative cell state by forming a heterodimer with Max, binding sequence-specific DNA elements and stimulating transcription of proliferation-associated genes. Transcription factors are key regulators of cell state as they control the gene expression programs that drive cell type specification and commonly are terminal components of a signaling cascade. These gene expression programs are often deregulated in disease states making transcription factors an ideal class of proteins for therapeutic targeting. However, most transcription factors lack clear pockets for small molecule binding and therefore have been largely considered undruggable with current technologies. A major challenge in the chemical biology field has been to develop potent small molecule inhibitors of transcription factors. While previously published work has identified small molecule inhibitors of c-Myc/Max heterodimerization using truncated proteins in FRET and yeast two-hybrid assays, only a few thousand compounds were screened in each case and the in vitro potency of these inhibitors is limited. Indeed, the potency fails to translate to in vivo activity in animal models. In general, these compounds fail to meet the generally agreed upon criteria for acceptable chemical probes. New chemotypes, which can be successfully developed into chemical probes, are desperately needed.)The proposed research aims to identify inhibitors of c-Myc/Max dimerization and DNA binding using novel technology. A robust high-throughput in vitro assay has been developed to screen for inhibitors of c-Myc/Max dimerization and subsequent binding to its DNA binding site. Secondary biochemical and cellular assays have been developed to validate hits from the primary screen and study them in greater detail. A large high-throughput screen as could be provided through the MLPCN using these robust assays should provide tractable hits for development and validation of biological effect. Medicinal chemistry optimizing these lead molecules through iterative use of downstream assays outlined herein then provides the opportunity to generate useful chemical probes to study c-Myc function. Such probes will hopefully lead the way to new therapeutics against this quintessential cancer target and offer insights into mechanisms for directly inhibiting transcription factors.

描述（由申请人提供）：MYC 是人类癌症中最常扩增的癌基因，发生在多种组织类型中，包括乳腺癌、肺癌和前列腺。 MYC 过度表达发生在至少 30% 的人类癌症中，并且经常与不良的临床结果和增加的复发机会相关。据估计，每年有 450,000 名美国人被诊断患有 MYC 依赖性癌症。这些患者需要新颖有效的治疗策略。 c-Myc 是一种螺旋-环-螺旋转录因子，通过与 Max 形成异二聚体、结合序列特异性 DNA 元件并刺激增殖相关基因的转录来驱动增殖细胞状态。转录因子是细胞状态的关键调节因子，因为它们控制驱动细胞类型规范的基因表达程序，并且通常是信号级联的终端组件。这些基因表达程序在疾病状态下通常会失调，从而使转录因子成为治疗靶向的理想蛋白质类别。然而，大多数转录因子缺乏用于小分子结合的清晰口袋，因此在很大程度上被认为用现有技术是不可成药的。化学生物学领域的一个主要挑战是开发有效的转录因子小分子抑制剂。虽然之前发表的工作已经在 FRET 和酵母双杂交测定中使用截短蛋白鉴定了 c-Myc/Max 异二聚化的小分子抑制剂，但每种情况下仅筛选了几千种化合物，并且这些抑制剂的体外效力有限。事实上，这种效力未能转化为动物模型的体内活性。一般来说，这些化合物不能满足可接受的化学探针的普遍商定的标准。迫切需要能够成功开发成化学探针的新化学型。）拟议的研究旨在利用新技术识别 c-Myc/Max 二聚化和 DNA 结合的抑制剂。一种强大的高通量体外测定方法已被开发出来，用于筛选 c-Myc/Max 二聚化抑制剂以及随后与其 DNA 结合位点结合的抑制剂。已开发出二次生化和细胞测定法来验证初级筛选的命中并对其进行更详细的研究。使用这些稳健的检测方法，通过 MLPCN 可以提供大型高通量筛选，这将为生物效应的开发和验证提供易于处理的结果。通过迭代使用本文概述的下游测定法来优化这些先导分子的药物化学提供了生成有用的化学探针来研究 c-Myc 功能的机会。此类探针有望为针对这一典型癌症靶点的新疗法开辟道路，并为直接抑制转录因子的机制提供见解。