Development of high throughput screening assays and pilot screen for inhibitors of metalloproteases meprin α and β.
Development of high throughput screening assays and pilot screen for inhibitors of metalloproteases meprin α and β.
复制标题
DOI：
10.1002/bip.22527
复制
发表时间：
2014-09
期刊：
影响因子：
2.9
通讯作者：
Minond, Dmitriy
中科院分区：
文献类型：
--
作者：
Madoux, Franck;Tredup, Claudia;Spicer, Timothy P.;Scampavia, Louis;Chase, Peter S.;Hodder, Peter S.;Fields, Gregg B.;Becker-Pauly, Christoph;Minond, Dmitriy
锌金属蛋白酶meprin α和meprin β与多种疾病有关，如纤维化、炎症和神经退行性变，然而，没有选择性的小分子抑制剂可以研究它们在这些过程中的作用。为了解决这一缺乏分子工具的问题，我们开发了高通量筛选（HTS）试验，以发现meprin α和meprin β的抑制剂，并筛选了一系列表征良好的药物制剂（LOPAC, n = 1,280种化合物）。两个化合物（PPNDS, NF449）证实了它们对meprin β的活性和选择性。动力学研究揭示了竞争性（PPNDS）和混合竞争性/非竞争性（NF449）抑制机制，表明结合发生在meprin β活性位点。PPNDS和NF449均表现出低纳摩尔IC50和Ki值，使它们成为迄今为止报道的最有效和选择性的meprin β抑制剂。这些结果证明了meprin α和β测定法能够识别选择性化合物并丢弃初步筛选的伪像。
Zinc metalloproteinases meprin α and meprin β are implicated in a variety of diseases, such as fibrosis, inflammation and neurodegeneration, however, there are no selective small molecule inhibitors that would allow to study their role in these processes. To address this lack of molecular tools we have developed high throughput screening (HTS) assays to enable discovery of inhibitors of both meprin α and meprin β and screened a collection of well characterized pharmaceutical agents (LOPAC, n = 1,280 compounds). Two compounds (PPNDS, NF449) confirmed their activity and selectivity for meprin β. Kinetic studies revealed competitive (PPNDS) and mixed competitive/non-competitive (NF449) inhibition mechanisms suggesting that binding occurs in meprin β active site. Both PPNDS and NF449 exhibited low nanomolar IC50 and Ki values making them the most potent and selective inhibitors of meprin β reported to the date. These results demonstrate the ability of meprin α and β assays to identify selective compounds and discard artifacts of primary screening.
登录
查看更多内容
影响因子：
4.8
作者：
Bien, Jessica;Jefferson, Tamara;Pietrzik, Claus U.
通讯作者：
Pietrzik, Claus U.
DOI：
10.1073/pnas.95.1.346
发表时间：
1998-01-06
影响因子：
11.1
作者：
Hohenegger, M;Waldhoer, M;Freissmuth, M
通讯作者：
Freissmuth, M
影响因子：
6.5
作者：
Becker-Pauly, Christoph;Hoewel, Markus;Stoecker, Walter
通讯作者：
Stoecker, Walter
影响因子：
3.7
作者：
Becker, C;Kruse, MN;Stöcker, W
通讯作者：
Stöcker, W
影响因子：
4.8
作者：
Bertenshaw, GP;Norcum, MT;Bond, JS
通讯作者：
Bond, JS