Ebsulfur as a potent scaffold for inhibition and labelling of New Delhi metallo-beta-lactamase-1 in vitro and in vivo
Ebsulfur as a potent scaffold for inhibition and labelling of New Delhi metallo-beta-lactamase-1 in vitro and in vivo
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Ebsulfur 作为体外和体内抑制和标记新德里金属-β-内酰胺酶-1 的有效支架
DOI：
10.1016/j.bioorg.2018.11.035
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发表时间：
2019
影响因子：
5.1
通讯作者：
Yang Kewu
中科院分区：
文献类型：
--
作者：
Su Jianpeng;Liu Jiayun;Chen Cheng;Zhang Yuejuan;Yang Kewu
由新德里金属-β-内酰胺酶（NDM-1）引起的超级细菌感染已发展成为一种新兴的威胁，由于NDM-1在病原菌之间穿梭，因此其标记和抑制已被证明具有挑战性。在这里，我们报告了一种有效的共价支架，ebsulfur，用于体外和体内靶向蛋白质。酶动力学研究表明，除1a-带1外，其余18个化合物对NDM-1有抑制作用，IC_（50）为0.16-9 μM，其中1g为最佳抑制剂，IC_（50）为0.16 μM，且呈剂量和时间依赖性。此外，这些ebsulfur有效地恢复了头孢唑啉对E的抗菌活性。结果表明，1g、1 i和1 n浓度下的抑菌效果最好，当浓度为16 μg/mL时，MIC降低了256倍。平衡透析研究表明，ebsulfur破坏了NDM-1活性中心的一个Zn（II）离子的配位。SDS-PAGE分析表明，该抑制剂与NDM-1的结合是共价结合。另外，在livingE中标记NDM-1。共聚焦显微成像显示了重组蛋白NDM-1在细胞内的实时分布变化过程。此外，测试了这些依布硫类化合物对L929小鼠成纤维细胞的细胞毒性，以及它们恢复抗生素对临床菌株E.大肠杆菌EC 08产NDM-1。本文提出的ebsulfur支架对于开发NDM-1的共价不可逆抑制剂以及在体外和体内标记靶点是有价值的。
The superbug infection caused by New Delhi metallo-β-lactamase (NDM-1) has grown into an emerging threat, labelling and inhibition of NDM-1 has proven challenging due to its shuttling between pathogenic bacteria. Here, we report a potent covalent scaffold, ebsulfur, for targeting the proteinin vitroandin vivo. Enzymatic kinetic study indicated that eighteen ebsulfurs gained except1a–band1finhibited NDM-1, exhibiting an IC50value ranging of 0.16–9 μM, and1gwas found to be the best, dose- and time-dependent inhibitor with an IC50of 0.16 μM. Also, these ebsulfurs effectively restored the antibacterial activity of cefazolin againstE. coliexpressing NDM-1, and the best effect was observed to be from1g,1iand1n, resulting in an 256-fold reduction in MIC of the antibiotic at a dose of 16 μg/mL. The equilibrium dialysis study implied that the ebsulfur disrupted the coordination of one Zn(II) ion at active site of NDM-1. Labelling of NDM-1 using a constructed fluorescent ebsulfurEbs-Rsuggested that the inhibitor covalently bound to the target through SDS-PAGE analysisin vitro. Also, labelling NDM-1 in livingE. colicells withEbs-Rby confocal microscopic imaging showed the real-time distribution change process of intracellular recombinant protein NDM-1. Moreover, the cytotoxicity of these ebsulfurs against L929 mouse fibroblastic cells was tested, and their capability to restore antibacterial activity of antibiotic against clinical strainsE. coliEC08 producing NDM-1 was determined. The ebsulfur scaffold proposed here is valuable for development of the covalent irreversible inhibitors of NDM-1, and also for labelling the targetin vitroandin vivo.