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。已经获得了许多与重组人NNMT相匹配的化合物的共晶结构，并用于指导药物化学工作。在该项目被化学生物学联盟（CBC）管理下的NCI实验治疗学（NExT）项目接受后，四种具有吸引人的药物样特性的强效和选择性三唑酮类似物已在单剂量小鼠药代动力学-药效学实验中进行了表征，两种化合物正在卵巢癌动物模型中进行概念有效性验证实验。正在进行的和未来的活动将集中在这些化合物和未来的类似物与卵巢癌标准护理治疗相结合的多种疗效和生存模型的进一步表征。

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 structures of a number of hit compounds with recombinant human NNMT have been obtained and been used to guide the medicinal chemistry efforts. After the project was accepted by the NCI Experimental Therapeutics (NExT) program under the management of Chemical Biology Consortium (CBC), four potent and selective triazolone analogs with attractive drug-like properties have been characterized in single-dose mouse pharmacokinetic-pharmacodynamic experiments and two compounds are being progressed towards proof-of-concept efficacy experiments in an ovarian cancer animal model. Ongoing and future activities will focus on the further characterization in multiple efficacy and survival models of both these compounds and future analogs in combination with the standard of care treatment for ovarian cancer.