Probing binding and cellular activity of pyrrolidinone and piperidinone small molecules targeting the urokinase receptor.
Probing binding and cellular activity of pyrrolidinone and piperidinone small molecules targeting the urokinase receptor.
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DOI：
10.1002/cmdc.201300340
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发表时间：
2013-12
期刊：
影响因子：
3.4
通讯作者：
Meroueh, Samy O.
中科院分区：
文献类型：
--
作者：
Mani, Timmy;Liu, Degang;Zhou, Donghui;Li, Liwei;Knabe, William Eric;Wang, Fang;Oh, Kyungsoo;Meroueh, Samy O.
关键词：
尿激酶受体（uPAR）是一种细胞表面蛋白，是促进癌细胞侵袭和转移的短暂和紧密的蛋白质相互作用的复杂网络的一部分。在这里，我们评估的结合和生物活性的一类新的吡咯烷酮和哌啶酮化合物，沿着与衍生物的先前确定的吡唑和丙胺化合物。竞争实验表明，化合物取代了荧光标记肽（AE 147-FAM），抑制常数Ki为6 ~ 63 μM。基于结构的计算药效团分析，随后进行了广泛的显式溶剂分子动力学模拟和自由能计算，表明吡唑基1a和哌啶酮基4采用不同的结合模式，尽管它们具有相似的二维结构。在细胞中，化合物1b和1f显示出对乳腺MDA-MB-231和胰腺导管腺癌（PDAC）细胞增殖的显著抑制，但4 b即使在100 μM的浓度下也没有表现出细胞毒性。1f以浓度依赖性方式损害MDA-MB-231的侵袭、粘附和迁移，而4 b仅抑制侵袭。1f以浓度依赖性方式抑制明胶酶（MMP-9）活性，而4 b显示无作用，表明抑制细胞侵袭的不同机制。信号传导研究进一步强调了这些差异，表明吡唑化合物完全抑制ERK磷酸化并损害HIF 1 α和NF-κB信号传导，而吡咯烷酮和哌啶酮（和4 B）没有影响。膜联蛋白V染色表明基于吡唑的1f对增殖的影响是由于通过凋亡机制杀死细胞。
The urokinase receptor (uPAR) is a cell-surface protein that is part of an intricate web of transient and tight protein interactions that promote cancer cell invasion and metastasis. Here we evaluate the binding and biological activity of a new class of pyrrolidinone and piperidinone compounds, along with derivatives of previously-identified pyrazole and propylamine compounds. Competition assays revealed that the compounds displaced a fluorescently-labeled peptide (AE147-FAM) with inhibition constant Ki ranging from 6 to 63 μM. Structure-based computational pharmacophore analysis followed by extensive explicit-solvent molecular dynamics simulations and free energy calculations suggested pyrazole-based 1a and piperidinone-based 4 adopt different binding modes, despite their similar two-dimensional structures. In cells, compounds 1b and 1f showed significant inhibition of breast MDA-MB-231 and pancreatic ductal adenocarcinoma (PDAC) cell proliferation, but 4b exhibited no cytotoxicity even at concentrations of 100 μM. 1f impaired MDA-MB-231 invasion, adhesion, and migration in a concentration-dependent manner, while 4b inhibited only invasion. 1f inhibited gelatinase (MMP-9) activity in a concentration-dependent manner, while 4b showed no effect suggesting different mechanisms for inhibition of cell invasion. Signaling studies further highlighted these differences, showing that pyrazole compounds completely inhibited ERK phosphorylation and impaired HIF1α and NF-κB signaling, while pyrrolidinone and piperidinone ( and 4b) had no effect. Annexin V staining suggested that the effect of pyrazole-based 1f on proliferation was due to cell killing through an apoptotic mechanism.
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影响因子：
4.8
作者：
Gardsvoll, Henrik;Jacobsen, Benedikte;Ploug, Michael
通讯作者：
Ploug, Michael
影响因子：
4.4
作者：
JORGENSEN, WL;CHANDRASEKHAR, J;KLEIN, ML
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KLEIN, ML
影响因子：
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作者：
Kunigal, Sateesh;Lakka, Sajani S.;Rao, Jasti S.
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Rao, Jasti S.
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作者：
Khanna M;Wang F;Jo I;Knabe WE;Wilson SM;Li L;Bum-Erdene K;Li J;W Sledge G;Khanna R;Meroueh SO
通讯作者：
Meroueh SO
影响因子：
4
作者：
Ahmad, Aamir;Kong, Dejuan;Wang, Zhiwei;Sarkar, Sanila. H.;Banerjee, Sanjeev;Sarkar, Fazlul H.
通讯作者：
Sarkar, Fazlul H.