title: Identification and characterization of the first class of potent bacterial acetyl-CoA carboxylase inhibitors with antibacterial activity.
abstract: The multisubunit acetyl-CoA carboxylase, which catalyzes the first committed step in fatty acid biosynthesis, is broadly conserved among bacteria. Its rate-limiting role in formation of fatty acids makes this enzyme an attractive target for the design of novel broad-spectrum antibacterials. However, no potent inhibitors have been discovered so far. This report describes the identification and characterization of highly potent bacterial acetyl-CoA carboxylase inhibitors with antibacterial activity for the first time. We demonstrate that pseudopeptide pyrrolidine dione antibiotics such as moiramide B inhibit the Escherichia coli enzyme at nanomolar concentrations. Moiramide B targets the carboxyltransferase reaction of this enzyme with a competitive inhibition pattern versus malonyl-CoA (K(i) value = 5 nm). Inhibition at nanomolar concentrations of the pyrrolidine diones is also demonstrated using recombinantly expressed carboxyltransferases from other bacterial species (Staphylococcus aureus, Streptococcus pneumoniae, and Pseudomonas aeruginosa). We isolated pyrrolidine dione-resistant strains of E. coli, S. aureus, and Bacillus subtilis, which contain mutations within the carboxyltransferase subunits AccA or AccD. We demonstrate that such mutations confer resistance to pyrrolidine diones. Inhibition values (IC(50)) of >100 microm regarding an eukaryotic acetyl-CoA carboxylase from rat liver indicate high selectivity of pyrrolidine diones for the bacterial multisubunit enzyme. The natural product moiramide B and synthetic analogues show broad-spectrum antibacterial activity. The knowledge of the target and the availability of facile assays using carboxyltransferases from different pathogens will enable evaluation of the antibacterial potential of the pyrrolidine diones as a promising antibacterial compound class acting via a novel mode of action.
authors.affiliation: Institute of Anti-infectives Research, Institute of Chemistry Research, and Institute of Cardiovascular Research, Pharma Research, Bayer HealthCare AG, D-42096 Wuppertal, Germany. christoph.freiberg@bayerhealthcare.com
authors.forename: Christoph
authors.initials: C
authors.lastname: Freiberg
authors.forename: Nina A
authors.initials: NA
authors.lastname: Brunner
authors.forename: Guido
authors.initials: G
authors.lastname: Schiffer
authors.forename: Thomas
authors.initials: T
authors.lastname: Lampe
authors.forename: Jens
authors.initials: J
authors.lastname: Pohlmann
authors.forename: Michael
authors.initials: M
authors.lastname: Brands
authors.forename: Martin
authors.initials: M
authors.lastname: Raabe
authors.forename: Dieter
authors.initials: D
authors.lastname: Häbich
authors.forename: Karl
authors.initials: K
authors.lastname: Ziegelbauer
mesh_terms: D000103:Acetyl-CoA Carboxylase / Q000037:antagonists & inhibitors*; D000577:Amides / Q000494:pharmacology*; D000595:Amino Acid Sequence; D000818:Animals; D000890:Anti-Infective Agents / Q000494:pharmacology*; D001412:Bacillus subtilis / Q000378:metabolism; D001419:Bacteria / Q000201:enzymology*; D001667:Binding, Competitive; D019731:Carbon-Nitrogen Ligases / Q000378:metabolism; D003001:Cloning, Molecular; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004926:Escherichia coli / Q000378:metabolism; D020128:Inhibitory Concentration 50; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D008954:Models, Biological; D008956:Models, Chemical; D008969:Molecular Sequence Data; D009154:Mutation; D017434:Protein Structure, Tertiary; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D013211:Staphylococcus aureus / Q000378:metabolism; D013388:Succinimides / Q000494:pharmacology*; D013997:Time Factors
chemical_list: D000577:Amides; D000890:Anti-Infective Agents; D004791:Enzyme Inhibitors; D011994:Recombinant Proteins; D013388:Succinimides; C487819:moiramide B; D019731:Carbon-Nitrogen Ligases; C019791:biotin carboxylase; D000103:Acetyl-CoA Carboxylase