Novel Inhibitors of Human N-Myristoyltransferase

该项目的目标是鉴定和表征作为癌症治疗剂有效的人N-肉豆蔻酰转移酶（NMT）的新型抑制剂。研究表明，NMT催化几种癌蛋白加工中的关键步骤，并且该过程的遗传抑制消除了致癌作用。NMT将肉豆蔻酰脂质连接到特定靶蛋白的N末端。这种不可逆的脂化使得蛋白质构象变化、膜缔合和进一步的翻译后加工成为可能，所有这些都赋予这些靶蛋白活性。此外，NMT在各种人类中过表达， 癌的这一发现，结合癌基因产物的必要加工，将NMT确定为潜在的抗癌治疗靶点。 尽管有越来越多的证据，但NMT的药理学抑制作为癌症治疗的一种手段仍然未被探索。为了解决这个问题，我们已经启动了一个项目，以发现和表征人类NMT的小分子抑制剂。通过开发一种新的筛选试验和测试合成化合物的库，我们确定了两种抑制NMT活性的化学型：环己基-八氢-吡咯并[1，2- a]吡嗪（COPP）和含金刚烷的化合物（ACC）。从文库中分离出共享这些化学型的化合物，并证明了体外和体内效力。为了对这些化学型的潜在效用进行原理性评价的证明，本研究将讨论以下具体目标 项目： 1.利用QSAR和计算酶对接研究设计和合成环己基-八氢-吡咯并[1，2-a]吡嗪和含金刚烷的化合物的类似物。 2.使用纯化的重组人NMT和基于细胞的测定来评估这些化合物。 3.确定先导NMT抑制剂的体内毒性、药代动力学和抗肿瘤活性。

The goal of this project is to identify and characterize novel inhibitors of human N-myristoyltransferase (NMT) that are effective as cancer therapeutic agents. Studies have shown that NMT catalyzes critical steps in the processing of several oncoproteins, and that genetic inhibition of this process ablates carcinogenesis. NMT attaches a myristoyl lipid to the N-terminus of specific target proteins. This irreversible lipidation enables protein conformational changes, membrane association, and further posttranslational processing, all of which confer activity to these target proteins. Furthermore, NMT is overexpressed in various human cancers. This finding, in conjunction with the necessary processing of oncogene products, identifies NMT as a potential therapeutic target against cancer. Despite this accumulating evidence, pharmacological inhibition of NMT as a means of cancer therapy remains unexplored. To address this problem, we have initiated a project to discover and characterize small molecule inhibitors of human NMT. By developing a novel screening assay and testing a library of synthetic compounds, we identified two chemotypes that inhibit NMT activity: cyclohexyl-octahydro-pyrrolo[1,2- a]pyrazine (COPP) and adamantine-containing compounds (ACC). Compounds sharing these chemotypes were isolated from the library and demonstrated in vitro and in vivo potency. To develop proof of principle evaluations of the potential utility of these chemotypes, the following Specific Aims will be addressed in this project: 1. Design and synthesize analogs of cyclohexyl-octahydro-pyrrolo[1,2-a]pyrazines and adamantine-containing compounds using QSAR and computational enzyme docking studies. 2. Evaluate these compounds using purified recombinant human NMT and cell-based assays. 3. Determine the in vivo toxicity, pharmacokinetics and antitumor activity of lead NMT inhibitors.