Inhibition of monoacylglycerol lipase by troglitazone, N-arachidonoyl dopamine and the irreversible inhibitor JZL184: comparison of two different assays
Inhibition of monoacylglycerol lipase by troglitazone, N-arachidonoyl dopamine and the irreversible inhibitor JZL184: comparison of two different assays
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DOI：
10.1111/j.1476-5381.2010.00974.x
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发表时间：
2010-12-01
影响因子：
7.3
通讯作者：
Fowler, C. J.
中科院分区：
文献类型：
--
作者：
Bjorklund, E.;Noren, E.;Fowler, C. J.
背景和临床上使用的PURPOSEDrugs通常有一个主要的作用机制，但对其他生物靶点的额外作用可能有助于它们的效果。一个潜在有用的额外靶点是内源性大麻素代谢酶单甘油脂肪酶(MGL)。我们筛选了一系列抑制MGL的药物，并比较了不同MGL酶分析的观察效价。以4-硝基苯乙酸酯(NPA)为底物，重组人MGL(细胞裂解产物和纯化酶)为底物，筛选了实验活性。在NPA法筛选的96个化合物中，曲格列酮、CP55、940、N-花生四烯基多巴胺和AM404对NPA裂解产物的IC50值分别为1.1、4.9、0.78和3.1mU·M。曲格列酮的效力与其主要药理活性相同，即激活过氧化物酶体增殖物激活受体(PPAR)γ。在PPARγ配体中，对人mgl的效价顺序为曲格列酮；环格列酮；罗格列酮；15-脱氧-三角洲(12，14)-前列腺素J(2)，接近Cay 10415；Gt；Cay 10514。与时间依赖的抑制剂JZL184不同，曲格列酮的效力取决于所使用的酶测定系统。因此，曲格列酮对大鼠胞浆中2-油酰甘油的抑制作用低于人MGL裂解酶对NPA的抑制作用(IC50为41µM)。结论MGL抑制剂筛选程序中的“HITS”应在不同的MGL检测中进行评估。曲格列酮可能是设计新型双作用MGL抑制剂/PPAR伽马激活剂的有用先导。
BACKGROUND AND PURPOSEDrugs used clinically usually have a primary mechanism of action, but additional effects on other biological targets can contribute to their effects. A potentially useful additional target is the endocannabinoid metabolizing enzyme monoacylglycerol lipase (MGL). We have screened a range of drugs for inhibition of MGL and compared the observed potencies using different MGL enzyme assays.EXPERIMENTAL APPROACHMGL activity was screened using recombinant human MGL (cell lysates and purified enzyme) with 4-nitrophenyl acetate (NPA) as substrate. 2-Oleolyglycerol metabolism by rat cerebellar cytosolic MGL and by recombinant MGL was also investigated.KEY RESULTSAmong the 96 compounds screened in the NPA assay, troglitazone, CP55,940, N-arachidonoyl dopamine and AM404 inhibited NPA hydrolysis by the lysates with IC50 values of 1.1, 4.9, 0.78 and 3.1 mu M, respectively. The potency for troglitazone is in the same range as its primary pharmacological activity, activation of peroxisome proliferator-activated receptor (PPAR) gamma. Among PPAR gamma ligands, the potency order towards human MGL was troglitazone > ciglitazone > rosiglitazone > 15-deoxy-Delta(12,14)-prostaglandin J(2) approximate to CAY 10415 > CAY 10514. In contrast to the time-dependent inhibitor JZL184, the potency of troglitazone was dependent upon the enzyme assay system used. Thus, troglitazone inhibited rat cytosolic 2-oleoylglycerol hydrolysis less potently (IC50 41 mu M) than hydrolysis of NPA by the human MGL lysates.CONCLUSIONS AND IMPLICATIONS'Hits' in screening programmes for MGL inhibitors should be assessed in different MGL assays. Troglitazone may be a useful lead for the design of novel, dual action MGL inhibitors/PPAR gamma activators.