qHTS to Identify Inhibitors of NNMT1

项目组成员已经确定了代谢酶NNMT在卵巢癌基质中的核心作用。靶酶在卵巢癌转移的基质中高度表达，也在原发性癌症相关成纤维细胞（CAFs）中表达。这种酶的敲低会导致caf的许多特征的逆转，并减弱它们在体外和体内促进癌细胞生长的能力。在此期间，项目团队成功优化了一种生化检测方法，用于筛选超过10万种化合物对NNMT的抑制活性。对两种检测成分和三种相关的甲基转移酶进行反筛选，以过滤掉假阳性。几种化学型被鉴定出具有良好的活性谱，并评估了体外靶标接合和细胞抑制作用。获得了其中一种与重组人NNMT共晶结构的化合物，并用于指导药物化学工作。目前正在对选定的药物进行优化，以提高效力和药动学性质。该项目已被化学生物学联盟（CBC）管理下的NCI实验治疗学（NExT）项目接受。

Members of the project team have identified a central role of the metabolic enzyme NNMT in the stroma of ovarian cancer. The target enzyme is highly expressed in the stroma of ovarian cancer metastases and is also expressed in primary cancer-associated fibroblasts (CAFs). Knockdown of the enzyme leads to a reversion of many of the features of CAFs and attenuates their ability to promote cancer cell growth both in vitro and in vivo. During this period, the project team successfully optimized a biochemical assay that was used to screen over 100,000 compounds for inhibitory activity against NNMT. Counter-screens against two components of the assay, and three related methyltransferase enzymes, were performed to filter out the false positives. Several chemotypes were identified with favorable activity profiles, and in vitro target engagement and cellular inhibition were assessed. Co-crystal structure of one of the hit compounds with recombinant human NNMT has been obtained and been used to guide the medicinal chemistry efforts. Optimization of selected hits is currently underway to improve potency and pharmacokinectic properties. The project has been accepted by the NCI Experimental Therapeutics (NExT) program under the management of Chemical Biology Consortium (CBC).