title: Discovery of inhibitors of 4'-phosphopantetheine adenylyltransferase (PPAT) to validate PPAT as a target for antibacterial therapy.
abstract: Inhibitors of 4'-phosphopantetheine adenylyltransferase (PPAT) were identified through high-throughput screening of the AstraZeneca compound library. One series, cycloalkyl pyrimidines, showed inhibition of PPAT isozymes from several species, with the most potent inhibition of enzymes from Gram-positive species. Mode-of-inhibition studies with Streptococcus pneumoniae and Staphylococcus aureus PPAT demonstrated representatives of this series to be reversible inhibitors competitive with phosphopantetheine and uncompetitive with ATP, binding to the enzyme-ATP complex. The potency of this series was optimized using structure-based design, and inhibition of cell growth of Gram-positive species was achieved. Mode-of-action studies, using generation of resistant mutants with targeted sequencing as well as constructs that overexpress PPAT, demonstrated that growth suppression was due to inhibition of PPAT. An effect on bacterial burden was demonstrated in mouse lung and thigh infection models, but further optimization of dosing requirements and compound properties is needed before these compounds can be considered for progress into clinical development. These studies validated PPAT as a novel target for antibacterial therapy.
authors.affiliation: Infection Bioscience, AstraZeneca R&D Boston, Waltham, Massachusetts, USA.
authors.forename: Boudewijn L M
authors.initials: BL
authors.lastname: de Jonge
authors.forename: Grant K
authors.initials: GK
authors.lastname: Walkup
authors.forename: Sushmita D
authors.initials: SD
authors.lastname: Lahiri
authors.forename: Hoan
authors.initials: H
authors.lastname: Huynh
authors.forename: Georg
authors.initials: G
authors.lastname: Neckermann
authors.forename: Luke
authors.initials: L
authors.lastname: Utley
authors.forename: Tory J
authors.initials: TJ
authors.lastname: Nash
authors.forename: Jesse
authors.initials: J
authors.lastname: Brock
authors.forename: Maryann
authors.initials: M
authors.lastname: San Martin
authors.forename: Amy
authors.initials: A
authors.lastname: Kutschke
authors.forename: Michele
authors.initials: M
authors.lastname: Johnstone
authors.forename: Valerie
authors.initials: V
authors.lastname: Laganas
authors.forename: Laurel
authors.initials: L
authors.lastname: Hajec
authors.forename: Rong-Fang
authors.initials: RF
authors.lastname: Gu
authors.forename: Haihong
authors.initials: H
authors.lastname: Ni
authors.forename: Brendan
authors.initials: B
authors.lastname: Chen
authors.forename: Kim
authors.initials: K
authors.lastname: Hutchings
authors.forename: Elise
authors.initials: E
authors.lastname: Holt
authors.forename: David
authors.initials: D
authors.lastname: McKinney
authors.forename: Ning
authors.initials: N
authors.lastname: Gao
authors.forename: Stephania
authors.initials: S
authors.lastname: Livchak
authors.forename: Jason
authors.initials: J
authors.lastname: Thresher
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D001667:Binding, Competitive; D018360:Crystallography, X-Ray; D055808:Drug Discovery; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D005260:Female; D008168:Lung / Q000187:drug effects / Q000382:microbiology; D051379:Mice; D008958:Models, Molecular; D009713:Nucleotidyltransferases / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D010204:Pantetheine / Q000031:analogs & derivatives / Q000737:chemistry; D011008:Pneumococcal Infections / Q000188:drug therapy / Q000382:microbiology; D018410:Pneumonia, Bacterial / Q000188:drug therapy / Q000382:microbiology; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology*; D013211:Staphylococcus aureus / Q000187:drug effects* / Q000201:enzymology / Q000254:growth & development; D013296:Streptococcus pneumoniae / Q000187:drug effects* / Q000201:enzymology / Q000254:growth & development; D013848:Thigh / Q000382:microbiology
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D004791:Enzyme Inhibitors; D054852:Small Molecule Libraries; D010204:Pantetheine; D009713:Nucleotidyltransferases; C022315:pantetheine-phosphate adenylyltransferase; C003129:4'-phosphopantetheine
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