title: Untargeted Metabolomics To Ascertain Antibiotic Modes of Action.
abstract: Deciphering the mode of action (MOA) of new antibiotics discovered through phenotypic screening is of increasing importance. Metabolomics offers a potentially rapid and cost-effective means of identifying modes of action of drugs whose effects are mediated through changes in metabolism. Metabolomics techniques also collect data on off-target effects and drug modifications. Here, we present data from an untargeted liquid chromatography-mass spectrometry approach to identify the modes of action of eight compounds: 1-[3-fluoro-4-(5-methyl-2,4-dioxo-pyrimidin-1-yl)phenyl]-3-[2-(trifluoromethyl)phenyl]urea (AZ1), 2-(cyclobutylmethoxy)-5'-deoxyadenosine, triclosan, fosmidomycin, CHIR-090, carbonyl cyanidem-chlorophenylhydrazone (CCCP), 5-chloro-2-(methylsulfonyl)-N-(1,3-thiazol-2-yl)-4-pyrimidinecarboxamide (AZ7), and ceftazidime. Data analysts were blind to the compound identities but managed to identify the target as thymidylate kinase for AZ1, isoprenoid biosynthesis for fosmidomycin, acyl-transferase for CHIR-090, and DNA metabolism for 2-(cyclobutylmethoxy)-5'-deoxyadenosine. Changes to cell wall metabolites were seen in ceftazidime treatments, although other changes, presumably relating to off-target effects, dominated spectral outputs in the untargeted approach. Drugs which do not work through metabolic pathways, such as the proton carrier CCCP, have no discernible impact on the metabolome. The untargeted metabolomics approach also revealed modifications to two compounds, namely, fosmidomycin and AZ7. An untreated control was also analyzed, and changes to the metabolome were seen over 4 h, highlighting the necessity for careful controls in these types of studies. Metabolomics is a useful tool in the analysis of drug modes of action and can complement other technologies already in use.
authors.affiliation: University of Glasgow, Glasgow, United Kingdom.
authors.forename: Isabel M
authors.initials: IM
authors.lastname: Vincent
authors.affiliation: AstraZeneca, Waltham, Massachusetts, USA.
authors.forename: David E
authors.initials: DE
authors.lastname: Ehmann
authors.affiliation: AstraZeneca, Waltham, Massachusetts, USA.
authors.forename: Scott D
authors.initials: SD
authors.lastname: Mills
authors.affiliation: AstraZeneca, Waltham, Massachusetts, USA.
authors.forename: Manos
authors.initials: M
authors.lastname: Perros
authors.affiliation: University of Glasgow, Glasgow, United Kingdom Michael.barrett@glasgow.ac.uk.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Barrett
mesh_terms: D000217:Acyltransferases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D000241:Adenosine / Q000378:metabolism / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000378:metabolism / Q000494:pharmacology*; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000378:metabolism / Q000494:pharmacology; D002442:Ceftazidime / Q000378:metabolism / Q000494:pharmacology; D002473:Cell Wall / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D002853:Chromatography, Liquid; D004269:DNA, Bacterial / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D004926:Escherichia coli / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D005578:Fosfomycin / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D006877:Hydroxamic Acids / Q000378:metabolism / Q000494:pharmacology; D013058:Mass Spectrometry; D055442:Metabolome / Q000187:drug effects*; D055432:Metabolomics; D009703:Nucleoside-Phosphate Kinase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D011743:Pyrimidines / Q000378:metabolism / Q000494:pharmacology; D013729:Terpenes / Q000037:antagonists & inhibitors / Q000378:metabolism; D013912:Threonine / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D014260:Triclosan / Q000378:metabolism / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; C525369:CHIR 090; D004269:DNA, Bacterial; D006877:Hydroxamic Acids; D011743:Pyrimidines; D013729:Terpenes; D005578:Fosfomycin; D013912:Threonine; D014260:Triclosan; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; C024640:fosmidomycin; D002442:Ceftazidime; D000217:Acyltransferases; D009703:Nucleoside-Phosphate Kinase; C020093:dTMP kinase; D000241:Adenosine
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