Targeting the <i>Mycobacterium tuberculosis</i> transpeptidase Ldt<sub>Mt2</sub> with cysteine-reactive inhibitors including ebselen

Munnik, Mariska de; Lohans, Christopher T.; Lang, Pauline A.; Langley, Gareth W.; Malla, Tika R.; Tumber, Anthony; Schofield, Christopher J.; Brem, Jürgen

doi:10.1039/c9cc04145a

Cited by 32 publications

(49 citation statements)

References 28 publications

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“…These conditions, which provided Z′ and S/B values of 0.82 and 8.1, respectively, are likely to be suitable for high‐throughput screening (HTS). However, to permit HTS, methods for quenching the reaction were sought, and a panel of cysteine reactants was assessed for their ability to react with Ldt Mt2 (unpublished data) . Ebselen, a known cysteine‐reactive reagent, was found to rapidly quench the reaction between Ldt Mt2 and 3 (Figure B).…”

Section: Resultsmentioning

confidence: 99%

A Fluorescence‐Based Assay for Screening β‐Lactams Targeting the Mycobacterium tuberculosis Transpeptidase Ldt_Mt2

et al. 2019

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Mycobacterium tuberculosis l,d‐transpeptidases (Ldts), which are involved in cell‐wall biosynthesis, have emerged as promising targets for the treatment of tuberculosis. However, an efficient method for testing inhibition of these enzymes is not currently available. We present a fluorescence‐based assay for LdtMt2, which is suitable for high‐throughput screening. Two fluorogenic probes were identified that release a fluorophore upon reaction with LdtMt2, thus making it possible to assess the availability of the catalytic site in the presence of inhibitors. The assay was applied to a panel of β‐lactam antibiotics and related inhibitors; the results validate observations that the (carba)penem subclass of β‐lactams are more potent Ldt inhibitors than other β‐lactam classes, though unexpected variations in potency were observed. The method will enable systematic structure–activity relationship studies on Ldts, thereby facilitating the identification of new antibiotics active against M. tuberculosis.

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Section: Resultsmentioning

confidence: 99%

A Fluorescence‐Based Assay for Screening β‐Lactams Targeting the Mycobacterium tuberculosis Transpeptidase Ldt_Mt2

et al. 2019

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“…By targeting cysteine residues in the active site from critical enzymes of Mycobacterium tuberculosis (antigen 85 complex; transpeptidase Ldt Mt2), ebselen and some derivatives were considered promising for treating tuberculosis (de Munnik et al 2019 ; Favrot et al 2013 ; Goins et al 2017 ).…”

Section: Pharmacology Of Organoselenium Compoundsmentioning

confidence: 99%

Toxicology and pharmacology of synthetic organoselenium compounds: an update

2021

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Here, we addressed the pharmacology and toxicology of synthetic organoselenium compounds and some naturally occurring organoselenium amino acids. The use of selenium as a tool in organic synthesis and as a pharmacological agent goes back to the middle of the nineteenth and the beginning of the twentieth centuries. The rediscovery of ebselen and its investigation in clinical trials have motivated the search for new organoselenium molecules with pharmacological properties. Although ebselen and diselenides have some overlapping pharmacological properties, their molecular targets are not identical. However, they have similar anti-inflammatory and antioxidant activities, possibly, via activation of transcription factors, regulating the expression of antioxidant genes. In short, our knowledge about the pharmacological properties of simple organoselenium compounds is still elusive. However, contrary to our early expectations that they could imitate selenoproteins, organoselenium compounds seem to have non-specific modulatory activation of antioxidant pathways and specific inhibitory effects in some thiol-containing proteins. The thiol-oxidizing properties of organoselenium compounds are considered the molecular basis of their chronic toxicity; however, the acute use of organoselenium compounds as inhibitors of specific thiol-containing enzymes can be of therapeutic significance. In summary, the outcomes of the clinical trials of ebselen as a mimetic of lithium or as an inhibitor of SARS-CoV-2 proteases will be important to the field of organoselenium synthesis. The development of computational techniques that could predict rational modifications in the structure of organoselenium compounds to increase their specificity is required to construct a library of thiol-modifying agents with selectivity toward specific target proteins.

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“… 12 Among the six compounds, ebselen is a clinical candidate with anti-inflammatory and anti-oxidant activities. In preclinical studies, ebselen was reported to react with cysteine residues from completely unrelated proteins including the C-terminal domain of the HIV-1 capsid, 15 the mycobacterium tuberculosis transpeptidase LdtMt2, 16 glutamate dehydrogenase, 17 clostridium difficile toxins TcdA and TcdB, 18 the mycobacterium tuberculosis (Mtb) antigen 85C enzyme, 19 the hepatitis C virus NS3 helicase, 20 and many others. Ebselen was also reported to induce protein unfolding for the insulin-degrading enzyme.…”

Section: Introductionmentioning

confidence: 99%

Ebselen, disulfiram, carmofur, PX-12, tideglusib, and shikonin are non-specific promiscuous SARS-CoV-2 main protease inhibitors

Townsend

et al. 2020

Preprint

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There is an urgent need for vaccines and antiviral drugs to combat the COVID-19 pandemic. Encouraging progress has been made in developing antivirals targeting SARS-CoV-2, the etiological agent of COVID-19. Among the drug targets being investigated, the viral main protease (Mpro) is one of the most extensively studied drug targets. Mpro is a cysteine protease that hydrolyzes the viral polyprotein at more than 11 sites and it is highly conserved among coronaviruses. In addition, Mpro has a unique substrate preference for glutamine in the P1 position. Taken together, it appears that Mpro inhibitors can achieve both broad-spectrum antiviral activity and a high selectivity index. Structurally diverse compounds have been reported as Mpro inhibitors, with several of which also showed antiviral activity in cell culture. In this study, we investigated the mechanism of action of six previously reported Mpro inhibitors, ebselen, disulfiram, tideglusib, carmofur, shikonin, and PX-12 using a consortium of techniques including FRET-based enzymatic assay, thermal shift assay, native mass spectrometry, cellular antiviral assays, and molecular dynamics simulations. Collectively, the results showed that the inhibition of Mpro by these six compounds is non-specific and the inhibition is abolished or greatly reduced with the addition of reducing reagent DTT. In the absence of DTT, these six compounds not only inhibit Mpro, but also a panel of viral cysteine proteases including SARS-CoV-2 papain-like protease, the 2Apro and 3Cpro from enterovirus A71 (EV-A71) and EV-D68. However, none of the compounds inhibits the viral replication of EV-A71 or EV-D68, suggesting that the enzymatic inhibition potency IC50 values obtained in the absence of DTT cannot be used to faithfully predict their cellular antiviral activity. Overall, we provide compelling evidence suggesting that ebselen, disulfiram, tideglusib, carmofur, shikonin, and PX-12 are non-specific SARS-CoV-2 Mpro inhibitors, and urge the scientific community to be stringent with hit validation.

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Targeting the Mycobacterium tuberculosis transpeptidase Ldt_Mt2 with cysteine-reactive inhibitors including ebselen

Abstract: Inhibitors targeting the conserved nucleophilic cysteine of the mycobacterial l,d-transpeptidases are a potential strategy for the treatment of tuberculosis.

Cited by 32 publications

References 28 publications

A Fluorescence‐Based Assay for Screening β‐Lactams Targeting the Mycobacterium tuberculosis Transpeptidase Ldt_Mt2

A Fluorescence‐Based Assay for Screening β‐Lactams Targeting the Mycobacterium tuberculosis Transpeptidase Ldt_Mt2

Toxicology and pharmacology of synthetic organoselenium compounds: an update

Ebselen, disulfiram, carmofur, PX-12, tideglusib, and shikonin are non-specific promiscuous SARS-CoV-2 main protease inhibitors

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Targeting the Mycobacterium tuberculosis transpeptidase LdtMt2 with cysteine-reactive inhibitors including ebselen

Abstract: Inhibitors targeting the conserved nucleophilic cysteine of the mycobacterial l,d-transpeptidases are a potential strategy for the treatment of tuberculosis.

Cited by 32 publications

References 28 publications

A Fluorescence‐Based Assay for Screening β‐Lactams Targeting the Mycobacterium tuberculosis Transpeptidase LdtMt2

A Fluorescence‐Based Assay for Screening β‐Lactams Targeting the Mycobacterium tuberculosis Transpeptidase LdtMt2

Toxicology and pharmacology of synthetic organoselenium compounds: an update

Ebselen, disulfiram, carmofur, PX-12, tideglusib, and shikonin are non-specific promiscuous SARS-CoV-2 main protease inhibitors

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Product

Resources

About

Targeting the Mycobacterium tuberculosis transpeptidase Ldt_Mt2 with cysteine-reactive inhibitors including ebselen

A Fluorescence‐Based Assay for Screening β‐Lactams Targeting the Mycobacterium tuberculosis Transpeptidase Ldt_Mt2

A Fluorescence‐Based Assay for Screening β‐Lactams Targeting the Mycobacterium tuberculosis Transpeptidase Ldt_Mt2