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Identification of inhibitors of PvdQ, an enzyme involved in the synthesis of the siderophore pyoverdine.

Wurst JM, Drake EJ, Theriault JR, Jewett IT, VerPlank L, Perez JR, Dandapani S, Palmer M, Moskowitz SM, Schreiber SL, Munoz B, Gulick AM - ACS Chem. Biol. (2014)

Bottom Line: Here, we describe the discovery of ML318, a biaryl nitrile inhibitor of PvdQ acylase.ML318 inhibits PvdQ in vitro (IC50 = 20 nM) by binding in the acyl-binding site, as confirmed by the X-ray crystal structure of PvdQ bound to ML318.Additionally, the PvdQ inhibitor is active in a whole cell assay, preventing pyoverdine production and limiting the growth of P. aeruginosa under iron-limiting conditions.

View Article: PubMed Central - PubMed

Affiliation: The Broad Institute , Cambridge, Massachusetts 02142, United States.

ABSTRACT
Pseudomonas aeruginosa produces the peptide siderophore pyoverdine, which is used to acquire essential Fe(3+) ions from the environment. PvdQ, an Ntn hydrolase, is required for the biosynthesis of pyoverdine. PvdQ knockout strains are not infectious in model systems, suggesting that disruption of siderophore production via PvdQ inhibition could be exploited as a target for novel antibacterial agents, by preventing cells from acquiring iron in the low iron environments of most biological settings. We have previously described a high-throughput screen to identify inhibitors of PvdQ that identified inhibitors with IC50 values of ∼100 μM. Here, we describe the discovery of ML318, a biaryl nitrile inhibitor of PvdQ acylase. ML318 inhibits PvdQ in vitro (IC50 = 20 nM) by binding in the acyl-binding site, as confirmed by the X-ray crystal structure of PvdQ bound to ML318. Additionally, the PvdQ inhibitor is active in a whole cell assay, preventing pyoverdine production and limiting the growth of P. aeruginosa under iron-limiting conditions.

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Synthesis ofbiaryl nitrile 4 from bromopyridine 5 andbenzyl nitrile 6.
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fig3: Synthesis ofbiaryl nitrile 4 from bromopyridine 5 andbenzyl nitrile 6.

Mentions: Based on scaffold 3, a panel of analogues was synthesized and tested for PvdQinhibition activity. The racemates of most analogues were convenientlysynthesized via arylations of benzylacetonitriles with halopyridines31 or alkylations with haloalkanes.32 (We note that separation of the enantiomerswas not successful due to racemization of the benzylic position atambient temperatures and neutral conditions.) A representative exampleof the arylation is shown for the synthesis of biaryl nitrile 4 (Figure 3). Coupling of 2-bromo-5-trifluoromethylpyridine 5 with 4-fluorobenzyl nitrile 6 using potassium tert-butoxide gave the desired biaryl nitrile 4 in 69% yield. Tetrazoles were synthesized from nitriles via [3 +2] cycloaddition and amino nitriles were synthesized from 4-chlorobenzaldehydeby a Strecker reaction (not shown).33,34


Identification of inhibitors of PvdQ, an enzyme involved in the synthesis of the siderophore pyoverdine.

Wurst JM, Drake EJ, Theriault JR, Jewett IT, VerPlank L, Perez JR, Dandapani S, Palmer M, Moskowitz SM, Schreiber SL, Munoz B, Gulick AM - ACS Chem. Biol. (2014)

Synthesis ofbiaryl nitrile 4 from bromopyridine 5 andbenzyl nitrile 6.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4215858&req=5

fig3: Synthesis ofbiaryl nitrile 4 from bromopyridine 5 andbenzyl nitrile 6.
Mentions: Based on scaffold 3, a panel of analogues was synthesized and tested for PvdQinhibition activity. The racemates of most analogues were convenientlysynthesized via arylations of benzylacetonitriles with halopyridines31 or alkylations with haloalkanes.32 (We note that separation of the enantiomerswas not successful due to racemization of the benzylic position atambient temperatures and neutral conditions.) A representative exampleof the arylation is shown for the synthesis of biaryl nitrile 4 (Figure 3). Coupling of 2-bromo-5-trifluoromethylpyridine 5 with 4-fluorobenzyl nitrile 6 using potassium tert-butoxide gave the desired biaryl nitrile 4 in 69% yield. Tetrazoles were synthesized from nitriles via [3 +2] cycloaddition and amino nitriles were synthesized from 4-chlorobenzaldehydeby a Strecker reaction (not shown).33,34

Bottom Line: Here, we describe the discovery of ML318, a biaryl nitrile inhibitor of PvdQ acylase.ML318 inhibits PvdQ in vitro (IC50 = 20 nM) by binding in the acyl-binding site, as confirmed by the X-ray crystal structure of PvdQ bound to ML318.Additionally, the PvdQ inhibitor is active in a whole cell assay, preventing pyoverdine production and limiting the growth of P. aeruginosa under iron-limiting conditions.

View Article: PubMed Central - PubMed

Affiliation: The Broad Institute , Cambridge, Massachusetts 02142, United States.

ABSTRACT
Pseudomonas aeruginosa produces the peptide siderophore pyoverdine, which is used to acquire essential Fe(3+) ions from the environment. PvdQ, an Ntn hydrolase, is required for the biosynthesis of pyoverdine. PvdQ knockout strains are not infectious in model systems, suggesting that disruption of siderophore production via PvdQ inhibition could be exploited as a target for novel antibacterial agents, by preventing cells from acquiring iron in the low iron environments of most biological settings. We have previously described a high-throughput screen to identify inhibitors of PvdQ that identified inhibitors with IC50 values of ∼100 μM. Here, we describe the discovery of ML318, a biaryl nitrile inhibitor of PvdQ acylase. ML318 inhibits PvdQ in vitro (IC50 = 20 nM) by binding in the acyl-binding site, as confirmed by the X-ray crystal structure of PvdQ bound to ML318. Additionally, the PvdQ inhibitor is active in a whole cell assay, preventing pyoverdine production and limiting the growth of P. aeruginosa under iron-limiting conditions.

Show MeSH
Related in: MedlinePlus