Combinatorial enzymatic assay for the screening of a new class of bacterial cell wall inhibitors.
We have developed a screening assay by thin-layer chromatography (TLC) to identify inhibitors for the bacterial essential enzymes MurA, -B, and -C. Libraries of compounds were synthesized using the mix-and-split combinatorial chemistry approach. Screening of the pooled compounds using the developed assay revealed the presence of many pools active in vitro. Pools of interest were tested for antibacterial activity. Individual molecules in the active pools were synthesized and retested with the TLC assay and with bacteria. We focused on the best five compounds for further analysis. They were tested for inhibition on each of the three enzymes separately, and showed no inhibition of MurA or MurB activity but were all inhibitors of MurC enzyme. This approach yielded interesting lead compounds for the development of novel antibacterial agents.
El Zoeiby
Ahmed
A
Centre de recherche sur la fonction, structure et ingénierie des protéines, Faculté de médecine, pavillon Charles-Eugène-Marchand, Université Laval, Sainte-Foy, Québec, G1K 7P4, Canada.
Beaumont
Mélanie
M
Dubuc
Eric
E
Sanschagrin
François
F
Voyer
Normand
N
Levesque
Roger C
RC
D000107:Acetylation; D019883:Alkyl and Aryl Transferases / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism; D001419:Bacteria / Q000187:drug effects*; D002237:Carbohydrate Dehydrogenases / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism; D002473:Cell Wall / Q000187:drug effects*; D002855:Chromatography, Thin Layer; D020650:Combinatorial Chemistry Techniques; D004269:DNA, Bacterial / Q000235:genetics; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000494:pharmacology; D004926:Escherichia coli / Q000187:drug effects; D006898:Hydroxylamines / Q000737:chemistry; D007202:Indicators and Reagents; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D008826:Microbial Sensitivity Tests; D010453:Peptide Synthases / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism; D011437:Propylamines / Q000737:chemistry; D011550:Pseudomonas aeruginosa / Q000187:drug effects; D012972:Sodium Hydroxide / Q000737:chemistry; D014538:Uridine Diphosphate N-Acetylmuramic Acid
D004269:DNA, Bacterial; D004791:Enzyme Inhibitors; D006898:Hydroxylamines; D007202:Indicators and Reagents; D011437:Propylamines; D014538:Uridine Diphosphate N-Acetylmuramic Acid; D012972:Sodium Hydroxide; D002237:Carbohydrate Dehydrogenases; C023215:UDP-N-acetylmuramate dehydrogenase; D019883:Alkyl and Aryl Transferases; C022566:UDP-N-acetylglucosamine 1-carboxyvinyltransferase; D010453:Peptide Synthases; C023229:UDP-N-acetylmuramoyl-alanine synthetase