title: Clinically relevant Gram-negative resistance mechanisms have no effect on the efficacy of MC-1, a novel siderophore-conjugated monocarbam.
abstract: The incidence of hospital-acquired infections with multidrug-resistant (MDR) Gram-negative pathogens is increasing at an alarming rate. Equally alarming is the overall lack of efficacious therapeutic options for clinicians, which is due primarily to the acquisition and development of various antibiotic resistance mechanisms that render these drugs ineffective. Among these mechanisms is the reduced permeability of the outer membrane, which prevents many marketed antibiotics from traversing this barrier. To circumvent this, recent drug discovery efforts have focused on conjugating a siderophore moiety to a pharmacologically active compound that has been designed to hijack the bacterial siderophore transport system and trick cells into importing the active drug by recognizing it as a nutritionally beneficial compound. MC-1, a novel siderophore-conjugated β-lactam that promotes its own uptake into bacteria, has exquisite activity against many Gram-negative pathogens. While the inclusion of the siderophore was originally designed to facilitate outer membrane penetration into Gram-negative cells, here we show that this structural moiety also renders other clinically relevant antibiotic resistance mechanisms unable to affect MC-1 efficacy. Resistance frequency determinations and subsequent characterization of first-step resistant mutants identified PiuA, a TonB-dependent outer membrane siderophore receptor, as the primary means of MC-1 entry into Pseudomonas aeruginosa. While the MICs of these mutants were increased 32-fold relative to the parental strain in vitro, we show that this resistance phenotype is not relevant in vivo, as alternative siderophore-mediated uptake mechanisms compensated for the loss of PiuA under iron-limiting conditions.
authors.affiliation: Antibacterials Research Unit, Pfizer Worldwide Research & Development, Groton, CT, USA.
authors.forename: Craig J
authors.initials: CJ
authors.lastname: McPherson
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Aschenbrenner
authors.forename: Brian M
authors.initials: BM
authors.lastname: Lacey
authors.forename: Kelly C
authors.initials: KC
authors.lastname: Fahnoe
authors.forename: Margaret M
authors.initials: MM
authors.lastname: Lemmon
authors.forename: Steven M
authors.initials: SM
authors.lastname: Finegan
authors.forename: Baswanth
authors.initials: B
authors.lastname: Tadakamalla
authors.forename: John P
authors.initials: JP
authors.lastname: O'Donnell
authors.forename: John P
authors.initials: JP
authors.lastname: Mueller
authors.forename: Andrew P
authors.initials: AP
authors.lastname: Tomaras
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D003001:Cloning, Molecular; D003428:Cross Infection / Q000382:microbiology; D024881:Drug Resistance, Bacterial / Q000235:genetics / Q000502:physiology*; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D006090:Gram-Negative Bacteria / Q000187:drug effects* / Q000235:genetics*; D051379:Mice; D018272:Porins / Q000235:genetics; D011550:Pseudomonas aeruginosa / Q000187:drug effects / Q000235:genetics; D018805:Sepsis / Q000188:drug therapy / Q000382:microbiology; D017262:Siderophores; D001618:beta-Lactamases / Q000096:biosynthesis / Q000235:genetics; D047090:beta-Lactams / Q000494:pharmacology*
chemical_list: D000900:Anti-Bacterial Agents; D001425:Bacterial Outer Membrane Proteins; C578528:MC-1 beta-lactam compound; D018272:Porins; D017262:Siderophores; D047090:beta-Lactams; D001618:beta-Lactamases