Structural and Kinetic Studies of the Potent Inhibition of Metallo-β-lactamases by 6-Phosphonomethylpyridine-2-carboxylates.
Structural and Kinetic Studies of the Potent Inhibition of Metallo-β-lactamases by 6-Phosphonomethylpyridine-2-carboxylates.
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DOI：
10.1021/acs.biochem.7b01299
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发表时间：
2018-03-27
期刊：
影响因子：
2.9
通讯作者：
Dmitrienko GI
中科院分区：
文献类型：
--
作者：
Hinchliffe P;Tanner CA;Krismanich AP;Labbé G;Goodfellow VJ;Marrone L;Desoky AY;Calvopiña K;Whittle EE;Zeng F;Avison MB;Bols NC;Siemann S;Spencer J;Dmitrienko GI
目前临床上没有金属-β-内酰胺酶（MBL）的抑制剂，MBL是水解β-内酰胺抗生素并赋予革兰氏阴性菌耐药性的酶。在这里，我们提出6-膦酰基甲基吡啶-2-羧酸酯（PMPC）作为亚类B1（IMP-1，Vim-2和NDM-1）和B3（L1）MBL的有效抑制剂。抑制遵循竞争性缓慢结合模型，无异构化步骤（IC 50值为0.3-7.2 μM; Ki值为0.03-1.5 μM）。最小抑菌浓度试验证明，在真核细胞保持活力的浓度下，β-内酰胺（美罗培南）对产MBL细菌（包括临床分离株）的活性增强。晶体结构揭示了抑制剂与B1（IMP-1）和B3（L1）MBL的前所未有的结合模式。在IMP-1中，结合并不取代亲核氢氧化物，PMPC羧酸盐和吡啶氮与双核金属位点的Zn 2离子密切相互作用（分别为2.3和2.7 μ m）。膦酸酯基团产生有限的相互作用，但距离亲核氢氧化物2.6 μ m。此外，与PMPC膦酸酯和吡啶N-C2 π-键以及亲核氢氧化物相互作用的水分子的存在表明PMPC作为其水合物结合到MBL活性位点。结合是显着不同的L1，与膦酸酯取代两个锌，形成一个单锌酶，和亲核的氢氧化物，同时也使多个相互作用与蛋白质主链和锌1。羧酸根和吡啶氮分别与Ser 221和-223相互作用（3 π距离）。PMPC的效力、低毒性、细胞活性和对进一步修饰的顺从性表明这些和类似的膦酸酯化合物可以进一步考虑用于未来的MBL抑制剂开发。
There are currently no clinically available inhibitors of metallo-β-lactamases (MBLs), enzymes that hydrolyze β-lactam antibiotics and confer resistance to Gram-negative bacteria. Here we present 6-phosphonomethylpyridine-2-carboxylates (PMPCs) as potent inhibitors of subclass B1 (IMP-1, VIM-2, and NDM-1) and B3 (L1) MBLs. Inhibition followed a competitive, slow-binding model without an isomerization step (IC50 values of 0.3–7.2 μM; Ki values of 0.03–1.5 μM). Minimum inhibitory concentration assays demonstrated potentiation of β-lactam (Meropenem) activity against MBL-producing bacteria, including clinical isolates, at concentrations at which eukaryotic cells remain viable. Crystal structures revealed unprecedented modes of binding of inhibitor to B1 (IMP-1) and B3 (L1) MBLs. In IMP-1, binding does not replace the nucleophilic hydroxide, and the PMPC carboxylate and pyridine nitrogen interact closely (2.3 and 2.7 Å, respectively) with the Zn2 ion of the binuclear metal site. The phosphonate group makes limited interactions but is 2.6 Å from the nucleophilic hydroxide. Furthermore, the presence of a water molecule interacting with the PMPC phosphonate and pyridine N–C2 π-bond, as well as the nucleophilic hydroxide, suggests that the PMPC binds to the MBL active site as its hydrate. Binding is markedly different in L1, with the phosphonate displacing both Zn2, forming a monozinc enzyme, and the nucleophilic hydroxide, while also making multiple interactions with the protein main chain and Zn1. The carboxylate and pyridine nitrogen interact with Ser221 and -223, respectively (3 Å distance). The potency, low toxicity, cellular activity, and amenability to further modification of PMPCs indicate these and similar phosphonate compounds can be further considered for future MBL inhibitor development.
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DOI：
10.1107/s090744491003982x
发表时间：
2011-04
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Evans PR
通讯作者：
Evans PR
影响因子：
7.3
作者：
Chen AY;Thomas PW;Stewart AC;Bergstrom A;Cheng Z;Miller C;Bethel CR;Marshall SH;Credille CV;Riley CL;Page RC;Bonomo RA;Crowder MW;Tierney DL;Fast W;Cohen SM
通讯作者：
Cohen SM
影响因子：
21.8
作者：
Brem, Juergen;van Berkel, Sander S.;Schofield, Christopher J.
通讯作者：
Schofield, Christopher J.
影响因子：
5.6
作者：
Fonseca, Fatima;Bromley, Elizabeth H. C.;Spencer, James
通讯作者：
Spencer, James
影响因子：
5.6
作者：
CHEN, CCH;RAHIL, J;HERZBERG, O
通讯作者：
HERZBERG, O