Inhibition of pseudolysin and thermolysin by hydroxamate-based MMP inhibitors

Adekoya, Olayiwola A.; Sjøli, Stian; Wuxiuer, Yimingjiang; Bilto, Irina; Marques, Sérgio M.; Santos, M. Amélia; Nuti, Elisa; Cercignani, Giovanni; Rossello, Armando; Winberg, Jan‐Olof; Sylte, Ingebrigt

doi:10.1016/j.ejmech.2014.10.009

Cited by 18 publications

(27 citation statements)

References 50 publications

(29 reference statements)

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“…The docking of LM2 and ML25 indicated that both orientations could be possible in thermolysin. As noted previously there are subtle differences between the active site of pseudolysin and thermolysin [33]. The S2 ' -subpocket of pseudolysin is bigger than that of thermolysin, and the aromatic ring system of LM2 and ML25 seem to obtain stable interactions with the S2' subpocket in pseudolysin and contribute to relative stable pseudolysin-inhibitor complexes.…”

Section: Strong Interaction With Amino Acids In the S2mentioning

confidence: 70%

“…As discussed in a recent paper, there are structural differences between the M4 enzymes and the MMPs in the region of these subpockets [33]. In MMP-2 and MMP-9, Leu83 and Tyr142 (MMP-2 numbering in the structure used for docking, the fibronectin-domain is replaced by a short peptide linker) are protruding into the binding pocket and contribute to a quite narrow access both to the S1'-and S2'-subpockets.…”

Section: Strong Interaction With Amino Acids In the S2mentioning

confidence: 99%

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Synthesis, experimental evaluation and molecular modelling of hydroxamate derivatives as zinc metalloproteinase inhibitors

Sjøli

Nuti

Camodeca

et al. 2016

European Journal of Medicinal Chemistry

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Enzymes of the M4 family of zinc-metalloproteinases are virulence factors secreted from grampositive or gram-negative bacteria, and putative drug targets in the treatment of bacterial infections. In order to have a therapeutic value such inhibitors should not interfere with endogenous zinc-metalloproteinases. In the present study we have synthesised a series of hydroxamate derivatives and validated the compounds as inhibitors of the M4 enzymes thermolysin and pseudolysin, and the endogenous metalloproteinases ADAM-17, MMP-2 and MMP-9 using experimental binding studies and molecular modelling. In general, the compounds are stronger inhibitors of the MMPs than of the M4 enzymes, however, an interesting exception is LM2. The compounds bound stronger to pseudolysin than to thermolysin, and the molecular modelling studies showed that occupation of the S2 ' subpocket by an aromatic group is favourable for strong interactions with pseudolysin.

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Section: Strong Interaction With Amino Acids In the S2mentioning

confidence: 70%

Section: Strong Interaction With Amino Acids In the S2mentioning

confidence: 99%

Synthesis, experimental evaluation and molecular modelling of hydroxamate derivatives as zinc metalloproteinase inhibitors

Sjøli

Nuti

Camodeca

et al. 2016

European Journal of Medicinal Chemistry

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“…A GST fusion tag does not affect the activity of native IMPI, allowing us to produce a functional recombinant IMPI fusion protein in E. coli which showed a stable and batch‐independent ability to inhibit thermolysin and purified PE, with a steeper descent than that of the chemical inhibitor phosphoramidon (Figure ). IMPI is more specific and more potent than other PE‐inhibitors such as those based on hydroxamate, metal‐chelating dipeptides, or small molecules . Kany et al .…”

Section: Discussionmentioning

confidence: 99%

The therapeutic potential of the insect metalloproteinase inhibitor against infections caused by Pseudomonas aeruginosa

Eisenhardt

Schlupp

Höfer

et al. 2018

Journal of Pharmacy and Pharmacology

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Objectives The objective of this study was to investigate the therapeutic potential of the insect metalloproteinase inhibitor (IMPI) from Galleria mellonella, the only known specific inhibitor of M4 metalloproteinases. Methods The fusion protein IMPI‐GST (glutathione‐S‐transferase) was produced by fermentation in Escherichia coli and was tested for its ability to inhibit the proteolytic activity of the M4 metalloproteinases thermolysin and Pseudomonas elastase (PE), the latter a key virulence factor of the wound‐associated and antibiotic‐resistant pathogen Pseudomonas aeruginosa. We also tested the ability of IMPI to inhibit the secretome (Sec) of a P. aeruginosa strain obtained from a wound. Key findings We found that IMPI‐GST inhibited thermolysin and PE in vitro and increased the viability of human keratinocytes exposed to Sec by inhibiting detachment caused by changes in cytoskeletal morphology. IMPI‐GST also improved the cell migration rate in an in vitro wound assay and reduced the severity of necrosis caused by Sec in an ex vivo porcine wound model. Conclusions The inhibition of virulence factors is a novel therapeutic approach against antibiotic resistant bacteria. Our results indicate that IMPI is a promising drug candidate for the treatment of P. aeruginosa infections.

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“…The final galardin concentrations in the assays varied from 2.42·10 −11 to 2.42·10 −5 M. The enzymatic reaction was started by the addition of Mca-RPPGFSAFK(Dpn)-OH (4 μM in assay), and the initial rate of the reaction was followed for 30 min at 37°C using the same wavelengths as with the Perkin Elmer fluorimeter as described above. The IC 50 values were determined from a dose response plot v i / v 0 vs log [Inhibitor] as described previously [ 56 ].…”

Section: Methodsmentioning

confidence: 99%

“…However, it is important that drugs targeting the bacterial enzymes not interfere with the function of the human MPs. Our focus for some time has been on MP inhibitors [ 56 , 57 ]. By studying the binding of various inhibitors to bacterial and human MPs, we are aiming to obtain information about similarities and differences in the active site of these enzymes that can be used in the development of compounds that bind specifically to the bacterial enzymes.…”

Section: Introductionmentioning

confidence: 99%

The selectivity of galardin and an azasugar-based hydroxamate compound for human matrix metalloproteases and bacterial metalloproteases

et al. 2018

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Inhibitors targeting bacterial enzymes should not interfere with enzymes of the host, and knowledge about structural determinants for selectivity is important for designing inhibitors with a therapeutic potential. We have determined the binding strengths of two hydroxamate compounds, galardin and compound 1b for the bacterial zinc metalloproteases, thermolysin, pseudolysin and auerolysin, known to be bacterial virulence factors, and the two human zinc metalloproteases MMP-9 and MMP-14. The active sites of the bacterial and human enzymes have huge similarities. In addition, we also studied the enzyme-inhibitor interactions by molecular modelling. The obtained Ki values of galardin for MMP-9 and MMP-14 and compound 1b for MMP-9 are approximately ten times lower than previously reported. Compound 1b binds stronger than galardin to both MMP-9 and MMP-14, and docking studies indicated that the diphenyl ether moiety of compound 1b obtains more favourable interactions within the S´1-subpocket than the 4-methylpentanoyl moiety of galardin. Both compounds bind stronger to MMP-9 than to MMP-14, which appears to be due to a larger S´1-subpocket in the former enzyme. Galardin, but not 1b, inhibits the bacterial enzymes, but the galardin Ki values were much larger than for the MMPs. The docking indicates that the S´1-subpockets of the bacterial proteases are too small to accommodate the diphenyl ether moiety of 1b, while the 4-methylpentanoyl moiety of galardin enters the pocket. The present study indicates that the size and shape of the ligand structural moiety entering the S´1-subpocket is an important determinant for selectivity between the studied MMPs and bacterial MPs.

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Inhibition of pseudolysin and thermolysin by hydroxamate-based MMP inhibitors

Cited by 18 publications

References 50 publications

Synthesis, experimental evaluation and molecular modelling of hydroxamate derivatives as zinc metalloproteinase inhibitors

Synthesis, experimental evaluation and molecular modelling of hydroxamate derivatives as zinc metalloproteinase inhibitors

The therapeutic potential of the insect metalloproteinase inhibitor against infections caused by Pseudomonas aeruginosa

The selectivity of galardin and an azasugar-based hydroxamate compound for human matrix metalloproteases and bacterial metalloproteases

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