Real‐Time Monitoring of New Delhi Metallo‐β‐Lactamase Activity in Living Bacterial Cells by <sup>1</sup>H NMR Spectroscopy

Ma, Junhe; McLeod, Sarah M.; MacCormack, Kathleen; Sriram, Shubha; Gao, Ning; Breeze, Alexander L.; Hu, Jun

doi:10.1002/ange.201308636

Cited by 15 publications

(8 citation statements)

References 37 publications

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“…36 Addition of the four BTZs inhibited imipenem hydrolysis by bacterial cells (Table 2 and Figure 2). L-CS319 was the most potent inhibitor in bacteria with an IC 50 value of 23 μ M, whereas its enantiomer was 4.7 times less active, a difference within the same order of magnitude as that observed in the in vitro experiments.…”

Section: Resultsmentioning

confidence: 95%

Bisthiazolidines: A Substrate-Mimicking Scaffold as an Inhibitor of the NDM-1 Carbapenemase

et al. 2015

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Pathogenic Gram-negative bacteria resistant to almost all β-lactam antibiotics are a major public health threat. Zn(II)-dependent or metallo-β-lactamases (MBLs) produced by these bacteria inactivate most β-lactam antibiotics, including the carbapenems, which are “last line therapies” for life-threatening Gram-negative infections. NDM-1 is a carbapenemase belonging to the MBL family that is rapidly spreading worldwide. Regrettably, inhibitors of MBLs are not yet developed. Here we present the bisthiazolidine (BTZ) scaffold as a structure with some features of β-lactam substrates, which can be modified with metal-binding groups to target the MBL active site. Inspired by known interactions of MBLs with β-lactams, we designed four BTZs that behave as in vitro NDM-1 inhibitors with Ki values in the low micromolar range (from 7 ± 1 to 19 ± 3 μM). NMR spectroscopy demonstrated that they inhibit hydrolysis of imipenem in NDM-1-producing Escherichia coli. In vitro time kill cell-based assays against a variety of bacterial strains harboring blaNDM-1 including Acinetobacter baumannii show that the compounds restore the antibacterial activity of imipenem. A crystal structure of the most potent heterocycle (L-CS319) in complex with NDM-1 at 1.9 Å resolution identified both structural determinants for inhibitor binding and opportunities for further improvements in potency.

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Section: Resultsmentioning

confidence: 95%

Bisthiazolidines: A Substrate-Mimicking Scaffold as an Inhibitor of the NDM-1 Carbapenemase

et al. 2015

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“…Besides the target proteins, NMR signals of small drugs can also be used to study proteindrug interactions in cells. For instance, hydrolysis of antibiotics by the New Delhi metallo-βlactamase subclass 1 (NDM-1) in E. coli was monitored in real time (62) (Figure 7). NDM-1 is one reason superbugs are resistant to most antibiotics (63).…”

Section: 22mentioning

confidence: 99%

“…). Ma et al(62) used one-dimensional (1D) 1 H NMR spectroscopy to monitor the hydrolytic decomposition of the carbapenem antibiotic meropenem in E. coli cells expressing NDM-1…”

mentioning

confidence: 99%

Magnetic Resonance Spectroscopy as a Tool for Assessing Macromolecular Structure and Function in Living Cells

Jie

Cheng

et al. 2017

Annual Rev. Anal. Chem.

214

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Investigating the structure, modification, interaction, and function of biomolecules in their native cellular environment leads to physiologically relevant knowledge about their mechanisms, which will benefit drug discovery and design. In recent years, nuclear and electron magnetic resonance (NMR) spectroscopy has emerged as a useful tool for elucidating the structure and function of biomacromolecules, including proteins, nucleic acids, and carbohydrates in living cells at atomic resolution. In this review, we summarize the progress and future of in-cell NMR as it is applied to proteins, nucleic acids, and carbohydrates.

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“…It is worth noting that the mercaptoacetic acid thioesters 1−4 and 7 significantly inhibit NDM-1 with an IC 50 value range of 12−96 μM, as pathogens expressing NDM-1 have been called superbugs. 24 Given the specificity of these mercaptoacetic acid thioesters targeting L1, in order to further identify the inhibition model of the inhibitors, we determined inhibition constants K i of the thioesters against L1. The data listed in Table 2 show that these compounds are very potent inhibitors with a K i value range of 0.11−0.95 μM.…”

Section: Acs Medicinal Chemistry Lettersmentioning

confidence: 99%

Amino Acid Thioester Derivatives: A Highly Promising Scaffold for the Development of Metallo-β-lactamase L1 Inhibitors

Liu

Shi

Kang

et al. 2015

ACS Med. Chem. Lett.

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In light of the biomedical significance of metallo-β-lactamases (MβLs), ten new mercaptoacetic acid thioester amino acid derivatives were synthesized and characterized. Biological activity assays indicated that all these synthesized compounds are very potent inhibitors of L1, exhibiting an IC 50 value range of 0.018−2.9 μM and a K i value range of 0.11−0.95 μM using cefazolin as substrate. Partial thioesters also showed effective inhibitory activities against NDM-1 and ImiS with an IC 50 value range of 12−96 and 3.6−65 μM, respectively. Also, all these thioesters increased susceptibility of E. coli cells expressing L1 to cefazolin, indicated by a 2−4-fold reduction in MIC of the antibiotic. Docking studies revealed potential binding modes of the two most potent L1 inhibitors to the active site in which the carboxylate group interacts with both Zn(II) ions and Ser221. This work introduces a highly promising scaffold for the development of metallo-β-lactamase L1 inhibitors. KEYWORDS: Antibiotic resistance, metallo-β-lactamase, subclass B3, L1, inhibitor, mercaptoacetic acid thioester β-Lactam antibiotics, which include penicillins, cephalosporins, and carbapenems, remain the most important and frequently used antimicrobial agents, constituting more than 50% of the antibiotics prescribed worldwide.1 However, the overuse of antibiotics in the clinical setting as well as animal production has resulted in resistance 2 conferred, among other resistance mechanisms, by the production of β-lactamases. More than a thousand β-lactamases have been isolated, and these enzymes have been categorized into classes A to D, depending on their amino acid sequence homologies.1 Enzymes from classes A, C, and D comprise the serine β-lactamases. They use a catalytic mechanism that is characterized by the nucleophilic attack of an active-site serine on the β-lactam carbonyl, ultimately causing cleavage of the β-lactam ring. Class B enzymes, or metallo-β-lactamases (MβLs), require either one or two Zn(II) ions per enzyme molecule for full catalytic activity, and these enzymes inactivate all clinically important β-lactams except aztreonam. According to amino acid sequence homologies MβLs are divided into subclasses B1−B3, based on amino acid sequence homologies.3 MβLs from subclasses B1 and B3 can inactivate nearly all β-lactam antibiotics. Contrarily, B2 enzymes possess a narrow substrate preference for carbapenems. There are no known clinical inhibitors of the MβLs to date.

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Real‐Time Monitoring of New Delhi Metallo‐β‐Lactamase Activity in Living Bacterial Cells by ¹H NMR Spectroscopy

Cited by 15 publications

References 37 publications

Bisthiazolidines: A Substrate-Mimicking Scaffold as an Inhibitor of the NDM-1 Carbapenemase

Bisthiazolidines: A Substrate-Mimicking Scaffold as an Inhibitor of the NDM-1 Carbapenemase

Magnetic Resonance Spectroscopy as a Tool for Assessing Macromolecular Structure and Function in Living Cells

Amino Acid Thioester Derivatives: A Highly Promising Scaffold for the Development of Metallo-β-lactamase L1 Inhibitors

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Real‐Time Monitoring of New Delhi Metallo‐β‐Lactamase Activity in Living Bacterial Cells by 1H NMR Spectroscopy

Cited by 15 publications

References 37 publications

Bisthiazolidines: A Substrate-Mimicking Scaffold as an Inhibitor of the NDM-1 Carbapenemase

Bisthiazolidines: A Substrate-Mimicking Scaffold as an Inhibitor of the NDM-1 Carbapenemase

Magnetic Resonance Spectroscopy as a Tool for Assessing Macromolecular Structure and Function in Living Cells

Amino Acid Thioester Derivatives: A Highly Promising Scaffold for the Development of Metallo-β-lactamase L1 Inhibitors

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Product

Resources

About

Real‐Time Monitoring of New Delhi Metallo‐β‐Lactamase Activity in Living Bacterial Cells by ¹H NMR Spectroscopy