Utilization of nitrophenylphosphates and oxime-based ligation for the development of nanomolar affinity inhibitors of the Yersinia pestis outer protein H (YopH) phosphatase

J Med Chem. 2011 Apr 28;54(8):2933-43. doi: 10.1021/jm200022g. Epub 2011 Mar 28.


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.

Publication types

  • Research Support, N.I.H., Intramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Bacteria / drug effects
  • Bacteria / growth & development
  • Bacterial Outer Membrane Proteins / antagonists & inhibitors*
  • Cell Line
  • Crystallography, X-Ray
  • Drug Design
  • Enzyme Inhibitors / chemistry*
  • Enzyme Inhibitors / pharmacology*
  • Enzyme Inhibitors / toxicity
  • Inhibitory Concentration 50
  • Mice
  • Models, Molecular
  • Oximes / chemistry*
  • Phosphates / chemistry*
  • Protein Tyrosine Phosphatases / antagonists & inhibitors*
  • Spectrometry, Mass, Electrospray Ionization
  • Yersinia pestis / enzymology*


  • Bacterial Outer Membrane Proteins
  • Enzyme Inhibitors
  • Oximes
  • Phosphates
  • Protein Tyrosine Phosphatases
  • yopH protein, Yersinia

Associated data

  • PDB/2Y2F