单酰基甘油脂肪酶(MAGL)参与调节促进癌症发病的脂肪酸网络系统，与多种癌症的发生密切相关，是一个潜在的抗癌新靶标。虽然已有一些抑制剂的报道，但多为共价不可逆性抑制剂，选择性低，副作用大。目前缺乏一种活性高、特异性强的可逆性MAGL小分子抑制剂。为此，我们通过构建酶活筛选体系，获得了一个结构新颖的先导化合物O7，并经过结构优化初步获得了活性高于O7的化合物。然而，这些化合物的活性并未达到预期，且选择性及体内活性也需进一步验证。因此本项目拟开展以下研究：以O7为基础继续进行优化，利用酶活测试和基于活性的蛋白质组学分析技术（ABPP）对化合物的生物活性及选择性进行评价；利用单晶X射线衍射探究化合物与蛋白的作用模式，进一步优化化合物结构，以获得高活性的靶向抑制剂；最后，对抑制剂的抗癌活性及分子机制进行研究。本项目的实施将有可能获得高活性的靶向MAGL抑制剂，为抗癌药物的研究奠定基础。

Monoacylglycerol lipase (MAGL) was identified as the principle lipolytic enzyme in cancer cells that produces free fatty acids from lipid precursors. It suggests that inhibition of MAGL constitutes a novel therapeutic avenue to treat cancer via interfering with a fatty acid network that promotes cancer pathogenesis. Currently, several MAGL inhibitors have been reported, but most of them belong to mechanism based covalent inhibitors, which induced several unwanted and side effects. Therefore, the development of highly potent and selective reversible MAGL inhibitors is essential. Previously, we established a natural substrate screening assay for MAGL, and obtained a lead compound O7 (IC50= 265 nM). After several rounds of optimization, we improved the potency of the compounds (~10-fold increase), but is not as good as expected. In this project, we would like to continue the optimization, and evaluate the potency and selectivity of the generated compounds by using natural substrate assay and a powerful technic named activity-based protein profiling (ABPP). Additionally, X-ray diffraction analysis will elucidate the mode of action that compounds bind to MAGL, which will aid our structure-based optimization. In the end, we plan to evaluate the anticancer activity of the optimized inhibitor. This project will contribute to the identification of MAGL inhibitors for cancer treatment.