qHTS to Identify Inhibitors of NNMT1

该项目组的成员已经确定了代谢酶NNMT在卵巢癌间质中的核心作用。靶酶在卵巢癌转移的基质中高度表达，并且也在原发性癌症相关成纤维细胞（CAF）中表达。敲除该酶导致CAF的许多特征逆转，并减弱其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物化学试验，用于筛选10万多种化合物对NNMT的抑制活性。对测定的两个组分和三种相关甲基转移酶进行反筛选，以滤出假阳性。鉴定了具有有利活性特征的几种化学型，并评估了体外靶标接合和细胞抑制。已获得其中一种命中化合物与重组人NNMT的共晶体结构，并用于指导药物化学工作。目前正在优化选定的命中，以提高效力和药代动力学特性。该项目已被化学生物学联盟（CBC）管理下的NCI实验治疗（NExT）计划接受。

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).