title: Inhibition of Neisseria gonorrhoeae Type II Topoisomerases by the Novel Spiropyrimidinetrione AZD0914.
abstract: We characterized the inhibition of Neisseria gonorrhoeae type II topoisomerases gyrase and topoisomerase IV by AZD0914 (AZD0914 will be henceforth known as ETX0914 (Entasis Therapeutics)), a novel spiropyrimidinetrione antibacterial compound that is currently in clinical trials for treatment of drug-resistant gonorrhea. AZD0914 has potent bactericidal activity against N. gonorrhoeae, including multidrug-resistant strains and key Gram-positive, fastidious Gram-negative, atypical, and anaerobic bacterial species (Huband, M. D., Bradford, P. A., Otterson, L. G., Basrab, G. S., Giacobe, R. A., Patey, S. A., Kutschke, A. C., Johnstone, M. R., Potter, M. E., Miller, P. F., and Mueller, J. P. (2014) In Vitro Antibacterial Activity of AZD0914: A New Spiropyrimidinetrione DNA Gyrase/Topoisomerase Inhibitor with Potent Activity against Gram-positive, Fastidious Gram-negative, and Atypical Bacteria. Antimicrob. Agents Chemother. 59, 467-474). AZD0914 inhibited DNA biosynthesis preferentially to other macromolecules in Escherichia coli and induced the SOS response to DNA damage in E. coli. AZD0914 stabilized the enzyme-DNA cleaved complex for N. gonorrhoeae gyrase and topoisomerase IV. The potency of AZD0914 for inhibition of supercoiling and the stabilization of cleaved complex by N. gonorrhoeae gyrase increased in a fluoroquinolone-resistant mutant enzyme. When a mutation, conferring mild resistance to AZD0914, was present in the fluoroquinolone-resistant mutant, the potency of ciprofloxacin for inhibition of supercoiling and stabilization of cleaved complex was increased greater than 20-fold. In contrast to ciprofloxacin, religation of the cleaved DNA did not occur in the presence of AZD0914 upon removal of magnesium from the DNA-gyrase-inhibitor complex. AZD0914 had relatively low potency for inhibition of human type II topoisomerases α and β.
authors.affiliation: Departments of Biosciences, Discovery Sciences, AstraZeneca R&D Boston, Waltham, Massachusetts 02451. Electronic address: gunther.kern@gmail.com.
authors.forename: Gunther
authors.initials: G
authors.lastname: Kern
authors.affiliation: Departments of Biosciences, Discovery Sciences, AstraZeneca R&D Boston, Waltham, Massachusetts 02451.
authors.forename: Tiffany
authors.initials: T
authors.lastname: Palmer
authors.affiliation: Departments of Biosciences, Discovery Sciences, AstraZeneca R&D Boston, Waltham, Massachusetts 02451.
authors.forename: David E
authors.initials: DE
authors.lastname: Ehmann
authors.affiliation: Departments of Biosciences, Discovery Sciences, AstraZeneca R&D Boston, Waltham, Massachusetts 02451.
authors.forename: Adam B
authors.initials: AB
authors.lastname: Shapiro
authors.affiliation: Departments of Biosciences, Discovery Sciences, AstraZeneca R&D Boston, Waltham, Massachusetts 02451.
authors.forename: Beth
authors.initials: B
authors.lastname: Andrews
authors.affiliation: Departments of Chemistry, Infection Innovative Medicines Unit, Discovery Sciences, AstraZeneca R&D Boston, Waltham, Massachusetts 02451.
authors.forename: Gregory S
authors.initials: GS
authors.lastname: Basarab
authors.affiliation: Department of Structure and Biophysics, Discovery Sciences, AstraZeneca R&D Boston, Waltham, Massachusetts 02451.
authors.forename: Peter
authors.initials: P
authors.lastname: Doig
authors.affiliation: Departments of Biosciences, Discovery Sciences, AstraZeneca R&D Boston, Waltham, Massachusetts 02451.
authors.forename: Jun
authors.initials: J
authors.lastname: Fan
authors.affiliation: Department of Structure and Biophysics, Discovery Sciences, AstraZeneca R&D Boston, Waltham, Massachusetts 02451.
authors.forename: Ning
authors.initials: N
authors.lastname: Gao
authors.affiliation: Departments of Biosciences, Discovery Sciences, AstraZeneca R&D Boston, Waltham, Massachusetts 02451.
authors.forename: Scott D
authors.initials: SD
authors.lastname: Mills
authors.affiliation: Departments of Biosciences, Discovery Sciences, AstraZeneca R&D Boston, Waltham, Massachusetts 02451.
authors.forename: John
authors.initials: J
authors.lastname: Mueller
authors.affiliation: Departments of Biosciences, Discovery Sciences, AstraZeneca R&D Boston, Waltham, Massachusetts 02451.
authors.forename: Shubha
authors.initials: S
authors.lastname: Sriram
authors.affiliation: Department of Structure and Biophysics, Discovery Sciences, AstraZeneca R&D Boston, Waltham, Massachusetts 02451.
authors.forename: Jason
authors.initials: J
authors.lastname: Thresher
authors.affiliation: Departments of Biosciences, Discovery Sciences, AstraZeneca R&D Boston, Waltham, Massachusetts 02451.
authors.forename: Grant K
authors.initials: GK
authors.lastname: Walkup
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001463:Barbiturates / Q000494:pharmacology*; D002939:Ciprofloxacin / Q000494:pharmacology; D002986:Clinical Trials as Topic; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D027081:DNA Gyrase / Q000235:genetics / Q000378:metabolism*; D027101:DNA Topoisomerase IV / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D004269:DNA, Bacterial / Q000737:chemistry* / Q000378:metabolism; D024881:Drug Resistance, Bacterial / Q000187:drug effects; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology / Q000235:genetics; D024841:Fluoroquinolones / Q000494:pharmacology; D015870:Gene Expression; D006801:Humans; D007555:Isoxazoles; D009025:Morpholines; D009154:Mutation; D009344:Neisseria gonorrhoeae / Q000187:drug effects / Q000201:enzymology / Q000235:genetics; D023303:Oxazolidinones; D013045:Species Specificity; D013141:Spiro Compounds / Q000494:pharmacology*; D059005:Topoisomerase II Inhibitors / Q000494:pharmacology*
chemical_list: D000900:Anti-Bacterial Agents; D001463:Barbiturates; D004269:DNA, Bacterial; D024841:Fluoroquinolones; D007555:Isoxazoles; D009025:Morpholines; D023303:Oxazolidinones; D013141:Spiro Compounds; D059005:Topoisomerase II Inhibitors; D002939:Ciprofloxacin; D004247:DNA; D027101:DNA Topoisomerase IV; D027081:DNA Gyrase; C000599190:zoliflodacin
keywords: AZD0914; DNA gyrase; DNA topoisomerase; antibiotic action; antibiotic resistance; enzyme inhibitor; gonorrhoeae