Novel Inhibitors of Human N-Myristoyltransferase

该项目的目标是确定和表征新型的人类N-肉豆蔻酰基转移酶(NMT)抑制剂，作为癌症治疗的有效药物。研究表明，NMT催化了几种癌蛋白加工的关键步骤，而对这一过程的遗传抑制可以消除癌症的发生。NMT将肉豆蔻基脂附着在特定靶蛋白的N端。这种不可逆的脂肪作用使蛋白质构象变化、膜结合和进一步的翻译后加工成为可能，所有这些都赋予了这些靶蛋白活性。此外，NMT在各种人类中都有过度表达 癌症。这一发现，结合对癌基因产物的必要处理，确定了NMT是一种潜在的癌症治疗靶点。 尽管有越来越多的证据，但NMT作为一种癌症治疗手段的药理抑制仍未被探索。为了解决这个问题，我们启动了一个发现和表征人类NMT小分子抑制剂的项目。通过建立一种新的筛选方法和测试合成化合物的文库，我们鉴定了两种抑制NMT活性的化学类型：环己基-八氢-吡咯并[1，2-a]吡嗪(CoPP)和含有金刚烷的化合物(ACC)。从文库中分离出具有这些化学类型的化合物，并在体外和体内证明了它们的有效性。为了对这些化学类型的潜在效用进行原则性评估，本报告将讨论以下具体目标 项目： 1.利用定量构效关系和计算机酶对接技术，设计合成了环己基-八氢吡咯并[1，2-a]吡嗪和金刚烷类化合物的类似物。 2.用纯化的重组人NMT和基于细胞的方法对这些化合物进行鉴定。 3.测定铅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.