Discovery of small molecule inhibitors for protein N-terminal methyltransferase

蛋白质 N 末端甲基转移酶小分子抑制剂的发现

基本信息

批准号：
9289669
负责人：
Rong Huang
金额：
$ 35.46万
依托单位：
PURDUE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-01 至 2020-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9289669
关键词：
Address Affinity Amination Amines Binding Biochemical Biological Biological Assay Biological Process Calorimetry Cancer Patient Cancer cell line Cell Proliferation Cell division Cell physiology Cells Chemicals Chromatin Collaborations Collection Colorectal Adenocarcinoma Colorectal Cancer Complex Coupled Coupling Crystallization DNA DNA Damage DNA Repair Defect Development Enzymes Fluorescence Fluorescent Dyes Foundations Functional disorder Genomics Goals Human Kinetics Knowledge Lead Libraries Link Malignant Neoplasms Mass Spectrum Analysis Measures Methylation Methyltransferase Mission Mitosis Mitotic Molecular Monitor N-terminal Normal tissue morphology Pathway interactions Peptides Pharmacologic Substance Phenotype Pilot Projects Play Production Promega Property Protein Methyltransferases Proteins Public Health Reproducibility Research Research Proposals Roentgen Rays Role Running S-Adenosylhomocysteine Series Specificity Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Structure System Technology Testing Therapeutic Tissues Titrations Translational Research X-Ray Crystallography assay development base cancer cell cell growth chemotherapeutic agent counterscreen design drug development drug discovery experience high throughput screening inhibitor/antagonist insight interest knock-down luminescence melanoma novel overexpression programs public health relevance response scaffold screening small hairpin RNA small molecule small molecule inhibitor small molecule libraries structural genomics success therapeutic target tumor progression tumorigenesis

项目摘要

Emerging evidence implies that protein methyltransferases (PMTs) that play a crucial role in diverse biological processes and various human dieases including cancer. Protein N-terminal methyltransferase 1 (NTMT1/NRMT1) plays an important role in cell division, DNA repair, and chromatin interaction. NTMT1 is upregulated in various cancer patients' tissues, and its over- expression level ranks in the top 1% in both colorectal adenocarcinoma and malignant melanoma. Moreover, loss of NTMT1 results in cancer cell mitotic defects and sensitizes the DNA to chemotherapeutic agents. Therefore, NTMT1 is a compelling target for development of novel cancer therapeutics for colorectal cancer and melanoma. Therefore, it is of interest to develop novel small molecule inhibitors that might serve as biological probes as well as lead molecules for drug development. However, there are no specific small molecule probes available for NTMT1 to provide new insights into the functions of NTMT1 methyltransferase activity in tumorigenesis. To fill this gap, our long-term goal is to discover of novel, potent, and selective small molecule NTMT1 inhibitors. For this application, we will develop a series of facile and reproducible high-throughput screening (HTS) assays and run a pilot screen comprised of about 15,000 compounds to validate these assays in collaboration with the National Center for Advancing Translational Sciences Chemical Genomics Center (NCGC). In Aim 1, we will develop primary, counter, and secondary HTS-compatible biochemical NTMT1 assays in a quantitative HTS format. In Aim 2, we will perform pilot studies with selected libraries to identify small molecule inhibitors for NTMT1. In aim 3, we will characterize active compounds in structural, selectivity, and cell-based studies. Upon completion of this project we expect to define biochemical and cell based assays to screen large libraries of small molecules to identify potent and selective inhibitors of NTMT1. We anticipate that these studies will result in the identification of potent and selective first-in- class NTMT1 small molecule inhibitors for further development as chemical probes. The knowledge gained would expedite the development of NTMT1 modulators and our understanding of NTMT1-regulated pathways in cancer patients. The NCGC's top tier screening program and our experience in drug discovery integrate the expertise needed to be successful in such endeavor.

新的证据表明蛋白质甲基转移酶（PMT）发挥着至关重要的作用不同的生物过程和包括癌症在内的各种人类疾病。蛋白质N末端甲基转移酶 1 (NTMT1/NRMT1) 在细胞分裂、DNA 修复和染色质相互作用。 NTMT1 在各种癌症患者的组织中表达上调，并且其过度表达在结直肠腺癌和恶性肿瘤中表达量均位于前1% 黑色素瘤。此外，NTMT1 的缺失会导致癌细胞有丝分裂缺陷，并使细胞变得敏感。 DNA 到化疗剂。因此，NTMT1是一个引人注目的开发目标针对结直肠癌和黑色素瘤的新型癌症疗法。因此，感兴趣的是开发新型小分子抑制剂，可作为生物探针和先导药物用于药物开发的分子。但目前尚无特异性小分子探针可用于 NTMT1，为 NTMT1 甲基转移酶的功能提供新的见解肿瘤发生中的活性。为了填补这一空白，我们的长期目标是发现新颖的、有效的、选择性小分子 NTMT1 抑制剂。对于这个应用，我们将开发一系列简便的和可重复的高通量筛选 (HTS) 测定，并运行包含以下内容的试点筛选：与国家中心合作验证了约 15,000 种化合物来验证这些测定推进转化科学化学基因组学中心 (NCGC)。在目标 1 中，我们将开发初级、计数器和次级 HTS 兼容的生化 NTMT1 检测定量 HTS 格式。在目标 2 中，我们将与选定的图书馆进行试点研究，以确定 NTMT1 的小分子抑制剂。在目标 3 中，我们将表征以下活性化合物：结构、选择性和基于细胞的研究。该项目完成后，我们预计将定义生化和细胞检测方法筛选大型小分子文库，以确定 NTMT1 的有效和选择性抑制剂。我们预计这些研究将导致确定有效且选择性的首创 NTMT1 类小分子抑制剂，进一步开发为化学探针。这所获得的知识将加速 NTMT1 调制器的开发和我们的了解癌症患者中 NTMT1 调节的通路。 NCGC 顶级筛选计划和我们在药物发现方面的经验整合了成功所需的专业知识在这样的努力中。