Discovery of Inhibitors of <i>Burkholderia pseudomallei</i> Methionine Aminopeptidase with Antibacterial Activity

Wangtrakuldee, Phumvadee; Byrd, Matthew S.; Campos, Cristine G.; Zhang, Zheng; Clare, Michael; Masoudi, Ali; Myler, Peter J.; Horn, James R.; Cotter, Peggy A.; Hagen, Timothy J.

doi:10.1021/ml400034m

Cited by 22 publications

(37 citation statements)

References 19 publications

(49 reference statements)

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“…The synthesis of the triazole species was previously reported 11 (Scheme 1). Briefly, the compounds were synthesized by base promoted addition of substituted benzyl bromides to the 5-amino-4H-1,2,4-triazole-3-thiol starting material.…”

Section: Resultsmentioning

confidence: 99%

“…For example, a nanomolar (IC 50 ) inhibitor of Burkholderia pseudomallei MetAP1 was found to exhibit growth inhibition of B. thailandensis . 11 In the present study, the inhibition activities of three distinct chemical classes of compounds (furoic acids, 1,2,4-triazoles and quinolinols) were evaluated against R. prowazekii MetAP1. Structure-activity relationship (SAR) analyses, along with molecular docking, were used to explore potential interactions of these inhibitors within the Rp MetAP active site.…”

Section: Introductionmentioning

confidence: 99%

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Rickettsia prowazekii methionine aminopeptidase as a promising target for the development of antibacterial agents

Helgren

Chen

Wangtrakuldee

et al. 2017

Bioorganic & Medicinal Chemistry

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Methionine aminopeptidase (MetAP) is a class of ubiquitous enzymes essential for the survival of numerous bacterial species. These enzymes are responsible for the cleavage of N-terminal formyl-methionine initiators from nascent proteins to initiate post-translational modifications that are often essential to proper protein function. Thus, inhibition of MetAP activity has been implicated as a novel antibacterial target. We tested this idea in the present study by targeting the MetAP enzyme in the obligate intracellular pathogen Rickettsia prowazekii. We first identified potent RpMetAP inhibitory species by employing an in vitro enzymatic activity assay. The molecular docking program AutoDock was then utilized to compare published crystal structures of inhibited MetAP species to docked poses of RpMetAP. Based on these in silico and in vitro screens, a subset of 23 compounds was tested for inhibition of R. prowazekii growth in a pulmonary vascular endothelial cell (EC) culture infection model system. All compounds were tested over concentration ranges that were determined to be non-toxic to the ECs and 10 of the 23 compounds displayed substantial inhibition of R. prowazekii growth. These data highlight the therapeutic potential for inhibiting RpMetAP as a novel antimicrobial strategy and set the stage for future studies in pre-clinical animal models of infection.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Rickettsia prowazekii methionine aminopeptidase as a promising target for the development of antibacterial agents

Helgren

Chen

Wangtrakuldee

et al. 2017

Bioorganic & Medicinal Chemistry

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“…Aryl carboxylic acid derivatives, usually 5-aryl-2-furoic acid or 5-aryl-2-thiophenic acids, have been demonstrated as potent inhibitors of bacterial MetAPs [19-22,33-43]. The compounds bind via chelation to one of the active site metals, as shown in the numerous crystal structures of these compounds bound to MetAPs from bacterial sources (PDB: 1XNZ, 2EVM, 2Q92, 2Q93, 2Q94, 2Q95, 2Q96 and 3IU7).…”

Section: Classes Of Metap Inhibitorsmentioning

confidence: 99%

“…Additionally, ( 17 ) was screened against Ab MetAPx and Ab MetAPy, with preferential binding again observed for the Mn(II) loaded forms of the enzymes ( Ab MetAPx IC 50 = 13, 394, 188, 358 μM; Mn(II), Co(II), Ni(II) and Fe(II), respectively; Ab MetAPy IC 50 = 0.92, 381, 186, 288; Mn(II), Co(II), Ni(II) and Fe(II), respectively) [21]. Compound ( 17 ) was also screened against Mt MetAP1c and was found to be active against the Mn(II) loaded form of the enzyme only ( Mt MetAP1c IC 50 = 14 μM; Mn(II); IC 50 = >500 μM; Co(II), Ni(II) and Fe(II)) [40] where inhibition of Bp MetAP1 with Co(II) cofactors by ( 17 ) was not observed (IC 50 = >250 μM) [22]. …”

Section: Classes Of Metap Inhibitorsmentioning

confidence: 99%

Advances in Bacterial Methionine Aminopeptidase Inhibition

Helgren¹,

Wangtrakuldee²,

Staker³

et al. 2015

CTMC

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Methionine aminopeptidases (MetAPs) are metalloenzymes that cleave the N-terminal methionine from newly synthesized peptides and proteins. These MetAP enzymes are present in bacteria, and knockout experiments have shown that MetAP activity is essential for cell life, suggesting that MetAPs are good antibacterial drug targets. MetAP enzymes are also present in the human host and selectivity is essential. There have been significant structural biology efforts and over 65 protein crystal structures of bacterial MetAPs are deposited into the PDB. This review highlights the available crystallographic data for bacterial MetAPs. Structural comparison of bacterial MetAPs with human MetAPs highlights differences that can lead to selectivity. In addition, this review includes the chemical diversity of molecules that bind and inhibit the bacterial MetAP enzymes. Analysis of the structural biology and chemical space of known bacterial MetAP inhibitors leads to a greater understanding of this antibacterial target and the likely development of potential antibacterial agents.

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“…Mutational studies have shown that the MetAP gene is essential for survival in bacteria [8,9] and cell proliferation in eukaryotes [10–12]. In various reports, inhibitors of MetAPs have been identified as potential anticancer [1,13,14], antimalarial [15,16], antibacterial [17,18] and anti-obesity [19,20] drugs. Due to the fact that bacteria contain a single distinct class (Type I) of the enzyme as opposed to those present in eukaryotes (Types I and II), there has been interest in designing isozyme selective inhibitors against bacterial MetAP as desirable antibiotic agents.…”

Section: Introductionmentioning

confidence: 99%

Bridging of a substrate between cyclodextrin and an enzyme's active site pocket triggers a unique mode of inhibition

Sule

Ugrinov

Mallik

et al. 2015

Biochimica et Biophysica Acta (BBA) - General Subjects

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Background Methionyl-7-amino-4-methylcoumarin (MetAMC) serves as a substrate for the E. coli Methionine aminopeptidase (MetAP) catalyzed reaction, and is routinely used for screening compounds to identify potential antibiotic agents. In pursuit of screening the enzyme’s inhibitors, we observed that 2-hydroxypropyl-β-cyclodextrin (HP-β-CD), utilized to solubilize hydrophobic inhibitors, inhibited the catalytic activity of the enzyme, and such inhibition was not solely due to sequestration of the substrate by HP-β-CD. Methods The mechanistic path for the HP-β-CD mediated inhibition of MetAP was probed by performing a detailed account of steady-state kinetics, ligand binding, X-ray crystallographic, and molecular modeling studies. Results X-ray crystallographic data of the β-cyclodextrin—substrate (β-CD—MetAMC) complex reveal that while the AMC moiety of the substrate is confined within the CD cavity, the methionine moiety protrudes outward. The steady-state kinetic data for inhibition of MetAP by HP-β-CD—MetAMC conform to a model mechanism in which the substrate is “bridged” between HP-β-CD and the enzyme’s active-site pocket, forming HP-β-CD—MetAMC—MetAP as the catalytically inactive ternary complex. Molecular modeling shows that the scissile bond of HP-β-CD-bound MetAMC substrate does not reach within the proximity of the enzyme’s catalytic metal center, and thus the substrate fails to undergo cleavage. Conclusions The data presented herein suggests that the bridging of the substrate between the enzyme and HP-β-CD cavities is facilitated by interaction of their surfaces, and the resulting complex inhibits the enzyme activity. General Significance Due to its potential interaction with physiological proteins via sequestered substrates, caution must be exercised in HP-β-CD mediated delivery of drugs under pathophysiological conditions.

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Discovery of Inhibitors of Burkholderia pseudomallei Methionine Aminopeptidase with Antibacterial Activity

Cited by 22 publications

References 19 publications

Rickettsia prowazekii methionine aminopeptidase as a promising target for the development of antibacterial agents

Rickettsia prowazekii methionine aminopeptidase as a promising target for the development of antibacterial agents

Advances in Bacterial Methionine Aminopeptidase Inhibition

Bridging of a substrate between cyclodextrin and an enzyme's active site pocket triggers a unique mode of inhibition

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