Scripps VIVO scripps research logo

  • Index
  • Log in
  • Home
  • People
  • Organizations
  • Research
  • Events
Search form
As of April 1st VIVO Scientific Profiles will no longer updated for faculty, and the link to VIVO will be removed from the library website. Faculty profile pages will continue to be updated via Interfolio. VIVO will continue being used behind the scenes to update graduate student profiles. Please contact helplib@scripps.edu if you have questions.
How to download citations from VIVO | Alternative profile options

Rational design of fatty acid amide hydrolase inhibitors that act by covalently bonding to two active site residues

Academic Article
uri icon
  • Overview
  • Identity
  • Additional Document Info
  • View All
scroll to property group menus

Overview

related to degree

  • Otrubova, Katerina, Ph.D. in Chemistry, Scripps Research 2007 - 2013

authors

  • Otrubova, Katerina
  • Brown, M.
  • McCormick, M. S.
  • Han, G. W.
  • O'Neal, S. T.
  • Cravatt, Benjamin
  • Stevens, Raymond
  • Lichtman, A. H.
  • Boger, Dale

publication date

  • April 2013

journal

  • Journal of the American Chemical Society  Journal

abstract

  • The design and characterization of ?-ketoheterocycle fatty acid amide hydrolase (FAAH) inhibitors are disclosed that additionally and irreversibly target a cysteine (Cys269) found in the enzyme cytosolic port while maintaining the reversible covalent Ser241 attachment responsible for their rapid and initially reversible enzyme inhibition. Two ?-ketooxazoles (3 and 4) containing strategically placed electrophiles at the C5 position of the pyridyl substituent of 2 (OL-135) were prepared and examined as inhibitors of FAAH. Consistent with the observed time-dependent noncompetitive inhibition, the cocrystal X-ray structure of 3 bound to a humanized variant of rat FAAH revealed that 3 was not only covalently bound to the active site catalytic nucleophile Ser241 as a deprotonated hemiketal, but also to Cys269 through the pyridyl C5-substituent, thus providing an inhibitor with dual covalent attachment in the enzyme active site. In vivo characterization of the prototypical inhibitors in mice demonstrates that they raise endogenous brain levels of FAAH substrates to a greater extent and for a much longer duration (>6 h) than the reversible inhibitor 2, indicating that the inhibitors accumulate and persist in the brain to completely inhibit FAAH for a prolonged period. Consistent with this behavior and the targeted irreversible enzyme inhibition, 3 reversed cold allodynia in the chronic constriction injury model of neuropathic pain in mice for a sustained period (>6 h) beyond that observed with the reversible inhibitor 2, providing effects that were unchanged over the 1-6 h time course monitored.
  • The design and characterization of α-ketoheterocycle fatty acid amide hydrolase (FAAH) inhibitors are disclosed that additionally and irreversibly target a cysteine (Cys269) found in the enzyme cytosolic port while maintaining the reversible covalent Ser241 attachment responsible for their rapid and initially reversible enzyme inhibition. Two α-ketooxazoles (3 and 4) containing strategically placed electrophiles at the C5 position of the pyridyl substituent of 2 (OL-135) were prepared and examined as inhibitors of FAAH. Consistent with the observed time-dependent noncompetitive inhibition, the cocrystal X-ray structure of 3 bound to a humanized variant of rat FAAH revealed that 3 was not only covalently bound to the active site catalytic nucleophile Ser241 as a deprotonated hemiketal, but also to Cys269 through the pyridyl C5-substituent, thus providing an inhibitor with dual covalent attachment in the enzyme active site. In vivo characterization of the prototypical inhibitors in mice demonstrates that they raise endogenous brain levels of FAAH substrates to a greater extent and for a much longer duration (>6 h) than the reversible inhibitor 2, indicating that the inhibitors accumulate and persist in the brain to completely inhibit FAAH for a prolonged period. Consistent with this behavior and the targeted irreversible enzyme inhibition, 3 reversed cold allodynia in the chronic constriction injury model of neuropathic pain in mice for a sustained period (>6 h) beyond that observed with the reversible inhibitor 2, providing effects that were unchanged over the 1-6 h time course monitored.

subject areas

  • Amidohydrolases
  • Animals
  • Binding, Competitive
  • Brain Chemistry
  • Catalytic Domain
  • Crystallography, X-Ray
  • Dose-Response Relationship, Drug
  • Drug Design
  • Enzyme Inhibitors
  • Escherichia coli
  • Hyperalgesia
  • Indicators and Reagents
  • Kinetics
  • Lipid Metabolism
  • Mice
  • Models, Molecular
  • Molecular Conformation
  • Neuralgia
  • Protein Carbonylation
  • Rats
  • Recombinant Proteins
  • Rhodamines
  • Substrate Specificity
scroll to property group menus

Identity

PubMed Central ID

  • PMC3678763

International Standard Serial Number (ISSN)

  • 1520-5126 (Electronic) 0002-7863 (Linking)

Digital Object Identifier (DOI)

  • 10.1021/ja4014997

PubMed ID

  • 23581831
scroll to property group menus

Additional Document Info

start page

  • 6289

end page

  • 6299

volume

  • 135

issue

  • 16

©2022 The Scripps Research Institute | Terms of Use | Powered by VIVO

  • About
  • Contact Us
  • Support