title: Distinguishing on-target versus off-target activity in early antibacterial drug discovery using a macromolecular synthesis assay.
abstract: The macromolecular synthesis assay was optimized in both S. aureus and E. coli imp and used to define patterns of inhibition of DNA, RNA, protein, and cell wall biosynthesis of several drug classes. The concentration of drug required to elicit pathway inhibition differed among the antimicrobial agents tested, with inhibition detected at concentrations significantly below the minimum inhibitory concentration (MIC) for tedizolid; within 4-fold of the MIC for ciprofloxacin, cefepime, vancomycin, tetracycline, and chloramphenicol; and significantly above the MIC for rifampicin and kanamycin. In a DNA gyrase/topoisomerase IV structure-based drug design optimization program, the assay rapidly identified undesirable off-target activity within certain chemotypes, altering the course of the program to focus on the series that maintained on-target activity.
authors.affiliation: 1Trius Therapeutics, Inc., San Diego, CA, USA.
authors.forename: Mark L
authors.initials: ML
authors.lastname: Cunningham
authors.forename: Bryan P
authors.initials: BP
authors.lastname: Kwan
authors.forename: Kirk J
authors.initials: KJ
authors.lastname: Nelson
authors.forename: Daniel C
authors.initials: DC
authors.lastname: Bensen
authors.forename: Karen J
authors.initials: KJ
authors.lastname: Shaw
mesh_terms: D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D001681:Biological Assay; D002473:Cell Wall / Q000187:drug effects; D027081:DNA Gyrase / Q000737:chemistry; D004269:DNA, Bacterial / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D055808:Drug Discovery; D004926:Escherichia coli / Q000187:drug effects* / Q000378:metabolism; D008826:Microbial Sensitivity Tests; D012329:RNA, Bacterial / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D013211:Staphylococcus aureus / Q000187:drug effects* / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D012329:RNA, Bacterial; D027081:DNA Gyrase
keywords: DNA gyrase; GyrB; ParE; macromolecular synthesis; pyrrolopyrimidine