Biochanin A, a naturally occurring inhibitor of fatty acid amide hydrolase.
OBJECTIVE
Inhibitors of fatty acid amide hydrolase (FAAH), the enzyme responsible for the metabolism of the endogenous cannabinoid (CB) receptor ligand anandamide (AEA), are effective in a number of animal models of pain. Here, we investigated a series of isoflavones with respect to their abilities to inhibit FAAH.


METHODS
In vitro assays of FAAH activity and affinity for CB receptors were used to characterize key compounds. In vivo assays used were biochemical responses to formalin in anaesthetized mice and the 'tetrad' test for central CB receptor activation.


RESULTS
Of the compounds tested, biochanin A was adjudged to be the most promising. Biochanin A inhibited the hydrolysis of 0.5 microM AEA by mouse, rat and human FAAH with IC(50) values of 1.8, 1.4 and 2.4 microM respectively. The compound did not interact to any major extent with CB(1) or CB(2) receptors, nor with FAAH-2. In anaesthetized mice, URB597 (30 microg i.pl.) and biochanin A (100 microg i.pl.) both inhibited the spinal phosphorylation of extracellular signal-regulated kinase produced by the intraplantar injection of formalin. The effects of both compounds were significantly reduced by the CB(1) receptor antagonist/inverse agonist AM251 (30 microg i.pl.). Biochanin A (15 mg.kg(-1) i.v.) did not increase brain AEA concentrations, but produced a modest potentiation of the effects of 10 mg.kg(-1) i.v. AEA in the tetrad test.


CONCLUSIONS
It is concluded that biochanin A, in addition to its other biochemical properties, inhibits FAAH both in vitro and peripherally in vivo.
Thors
L
L
Department of Pharmacology and Clinical Neuroscience, Umeå University, Umeå, Sweden.
Burston
J J
JJ
Alter
B J
BJ
McKinney
M K
MK
Cravatt
B F
BF
Ross
R A
RA
Pertwee
R G
RG
Gereau
R W
RW
Wiley
J L
JL
Fowler
C J
CJ
D000581:Amidohydrolases / Q000037:antagonists & inhibitors*; D000818:Animals; D001095:Arachidonic Acids / Q000378:metabolism; D001522:Behavior, Animal / Q000187:drug effects; D001549:Benzamides / Q000037:antagonists & inhibitors / Q000494:pharmacology; D001921:Brain / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D019556:COS Cells; D063387:Cannabinoid Receptor Antagonists; D002219:Carbamates / Q000037:antagonists & inhibitors / Q000494:pharmacology; D002461:Cell Line, Transformed; D002522:Chlorocebus aethiops; D004347:Drug Interactions; D063388:Endocannabinoids; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005557:Formaldehyde / Q000037:antagonists & inhibitors; D019833:Genistein / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D006801:Humans; D007529:Isoflavones / Q000494:pharmacology*; D008099:Liver / Q000201:enzymology; D051379:Mice; D008813:Mice, Inbred ICR; D010880:Piperidines / Q000494:pharmacology; D053284:Polyunsaturated Alkamides / Q000378:metabolism; D011720:Pyrazoles / Q000494:pharmacology; D051381:Rats; D000099252:Fatty Acid Amide Hydrolases
D000581:Amidohydrolases; D001095:Arachidonic Acids; D001549:Benzamides; D063387:Cannabinoid Receptor Antagonists; D002219:Carbamates; D063388:Endocannabinoids; D048049:Extracellular Signal-Regulated MAP Kinases; D005557:Formaldehyde; D019833:Genistein; D007529:Isoflavones; D010880:Piperidines; D053284:Polyunsaturated Alkamides; D011720:Pyrazoles; D000099252:Fatty Acid Amide Hydrolases; C500528:cyclohexyl carbamic acid 3'-carbamoylbiphenyl-3-yl ester; C103505:AM 251; C004541:biochanin A; C078814:anandamide