Discovery of small molecule inhibitors for protein N-terminal methyltransferase

新出现的证据表明，蛋白质甲基转移酶（PMT），发挥关键作用， 在不同的生物过程和包括癌症在内的各种人类疾病中。蛋白n端 甲基转移酶1（NTMT1/NRMT1）在细胞分裂、DNA修复和细胞凋亡中起重要作用。 染色质相互作用NTMT1在各种癌症患者的组织中上调，其过度表达 表达水平在大肠腺癌和恶性肿瘤中均居前1%， 黑素瘤此外，NTMT1的缺失导致癌细胞有丝分裂缺陷，并使肿瘤细胞对NTMT1敏感。 DNA与化疗药物的关系。因此，NTMT 1是开发的一个引人注目的目标， 用于结肠直肠癌和黑色素瘤的新型癌症治疗剂。因此，有必要 开发新的小分子抑制剂，可以作为生物探针， 用于药物开发的分子。然而，没有特异性的小分子探针 NTMT1甲基转移酶的功能提供了新的见解 在肿瘤发生中的活性。为了填补这一空白，我们的长期目标是发现新的，有效的， 选择性小分子NTMT1抑制剂。对于此应用程序，我们将开发一系列简便的 和可再现的高通量筛选（HTS）测定，并进行中试筛选， 大约15，000种化合物，以验证这些与国家中心合作的检测方法。 推进转化科学化学基因组学中心（NCGC）。在目标1中，我们 开发一级、计数器和二级HTS兼容的生化NTMT1检测试剂盒， 量化HTS格式。在目标2中，我们将对选定的图书馆进行试点研究， NTMT1的小分子抑制剂。在目标3中，我们将表征活性化合物， 结构、选择性和细胞研究。 在这个项目完成后，我们预计将确定生化和细胞为基础的测定 筛选大量的小分子文库，以鉴定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的和选择性的第一次在 NTMT1类小分子抑制剂，用于进一步开发作为化学探针。的 所获得的知识将加快NTMT1调节剂的开发， 了解NTMT1在癌症患者中的调节途径。NCGC的顶级筛选 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。

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.