title: Utilization of nitrophenylphosphates and oxime-based ligation for the development of nanomolar affinity inhibitors of the Yersinia pestis outer protein H (YopH) phosphatase.
abstract: Our current study reports the first K(M) optimization of a library of nitrophenylphosphate-containing substrates for generating an inhibitor lead against the Yersinia pestis outer protein phosphatase (YopH). A high activity substrate identified by this method (K(M) = 80 μM) was converted from a substrate into an inhibitor by replacement of its phosphate group with difluoromethylphosphonic acid and by attachment of an aminooxy handle for further structural optimization by oxime ligation. A cocrystal structure of this aminooxy-containing platform in complex with YopH allowed the identification of a conserved water molecule proximal to the aminooxy group that was subsequently employed for the design of furanyl-based oxime derivatives. By this process, a potent (IC(50) = 190 nM) and nonpromiscuous inhibitor was developed with good YopH selectivity relative to a panel of phosphatases. The inhibitor showed significant inhibition of intracellular Y. pestis replication at a noncytotoxic concentration. The current work presents general approaches to PTP inhibitor development that may be useful beyond YopH.
authors: Chemical Biology Laboratory, Molecular Discovery Program, Center for Cancer Research, National Cancer Institute, National Institutes of Health, NCI-Frederick, Frederick, Maryland 21702, United States.
authors: Medhanit
authors: M
authors: Bahta
authors: George T
authors: GT
authors: Lountos
authors: Beverly
authors: B
authors: Dyas
authors: Sung-Eun
authors: SE
authors: Kim
authors: Robert G
authors: RG
authors: Ulrich
authors: David S
authors: DS
authors: Waugh
authors: Terrence R
authors: TR
authors: Burke
mesh_terms: D000818:Animals; D001419:Bacteria / Q000187:drug effects / Q000254:growth & development; D001425:Bacterial Outer Membrane Proteins / Q000037:antagonists & inhibitors*; D002460:Cell Line; D018360:Crystallography, X-Ray; D015195:Drug Design; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000494:pharmacology* / Q000633:toxicity; D020128:Inhibitory Concentration 50; D051379:Mice; D008958:Models, Molecular; D010091:Oximes / Q000737:chemistry*; D010710:Phosphates / Q000737:chemistry*; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors*; D021241:Spectrometry, Mass, Electrospray Ionization; D015010:Yersinia pestis / Q000201:enzymology*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D004791:Enzyme Inhibitors; D010091:Oximes; D010710:Phosphates; D017027:Protein Tyrosine Phosphatases; C067571:yopH protein, Yersinia