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Rational Design of Fatty Acid Amide Hydrolase Inhibitors That Act by Covalently Bonding to Two Active Site Residues
- Source :
- Journal of the American Chemical Society. 135:6289-6299
- Publication Year :
- 2013
- Publisher :
- American Chemical Society (ACS), 2013.
-
Abstract
- 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 non-competitive inhibition, the co-crystal 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 demonstrate 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.
- Subjects :
- Models, Molecular
Stereochemistry
Molecular Conformation
Crystallography, X-Ray
Binding, Competitive
Biochemistry
Article
Catalysis
Amidohydrolases
Substrate Specificity
Protein Carbonylation
Mice
Colloid and Surface Chemistry
Non-competitive inhibition
Fatty acid amide hydrolase
Catalytic Domain
Escherichia coli
Animals
Enzyme Inhibitors
Brain Chemistry
chemistry.chemical_classification
Dose-Response Relationship, Drug
biology
Rhodamines
Rational design
Active site
General Chemistry
Lipid Metabolism
Recombinant Proteins
Rats
Kinetics
Enzyme
chemistry
Hyperalgesia
Covalent bond
Drug Design
Electrophile
biology.protein
Neuralgia
Indicators and Reagents
Cysteine
Subjects
Details
- ISSN :
- 15205126 and 00027863
- Volume :
- 135
- Database :
- OpenAIRE
- Journal :
- Journal of the American Chemical Society
- Accession number :
- edsair.doi.dedup.....476fd4bcfe814510f821bfebf7ab48bf