Characterization of the enzymatic activity for biphasic competition by guanoxabenz (1-(2,6-dichlorobenzylidene-amino)-3-hydroxyguanidine) at alpha2-adrenoceptors. II. Description of a xanthine-dependent enzymatic activity in spleen cytosol.
The mechanism for formation of high affinity binding of guanoxabenz (1-(2,6-dichlorobenzylidene-amino)-3-hydroxyguanidine) to alpha2-adrenoceptors by the rat spleen cytosol was studied. We report here that the spleen cytosolic fraction mediated the reduction of guanoxabenz to guanabenz (1-(2,6-dichlorobenzylidene-amino)-3-guanidine), the latter having an almost 100-fold higher affinity for rat alpha2A-adrenoceptors than guanoxabenz itself. The reaction product could be separated by high-performance liquid chromatography and its identity as guanabenz confirmed by nuclear magnetic resonance. The spleen cytosolic activity could be separated into high and low molecular weight components, the high molecular weight component requiring low molecular weight factors for maximal activity. Xanthine oxidase seems to be the most likely candidate responsible for the activity, as the guanoxabenz-reducing activity of the high molecular weight component could be sustained by exogenously applied xanthine, while it was potently blocked by allopurinol. The conversion of guanoxabenz by the cytosolic activity was also quite potently blocked by DWO1, 1-(3,4-dimethoxybenzylideneamino)3-hydroxyguanidine, a hydroxyguanidine analogue to guanoxabenz.
Dambrova
M
M
Department of Pharmaceutical Biosciences, Uppsala University, Sweden.
Uhlén
S
S
Welch
C J
CJ
Prusis
P
P
Wikberg
J E
JE
D000493:Allopurinol / Q000494:pharmacology; D000818:Animals; D000959:Antihypertensive Agents / Q000378:metabolism*; D001667:Binding, Competitive; D003600:Cytosol / Q000201:enzymology; D006143:Guanabenz / Q000031:analogs & derivatives* / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D018341:Receptors, Adrenergic, alpha-2 / Q000378:metabolism*; D013154:Spleen / Q000201:enzymology*; D019820:Xanthine / Q000494:pharmacology; D014969:Xanthine Oxidase / Q000502:physiology
D000959:Antihypertensive Agents; D018341:Receptors, Adrenergic, alpha-2; D019820:Xanthine; D000493:Allopurinol; D014969:Xanthine Oxidase; D006143:Guanabenz; C028782:guanoxabenz