Microsomal cytochrome P450 dependent oxidation of N-hydroxyguanidines, amidoximes, and ketoximes: mechanism of the oxidative cleavage of their C=N(OH) bond with formation of nitrogen oxides.
Oxidation by rat liver microsomes of 13 compounds involving a C=N(OH) function (including N-hydroxyguanidines, amidoximes, ketoximes, and aldoximes) was found to occur with the release of nitrogen oxides such as NO, NO2-, and NO3-. The greatest activities were observed with liver microsomes from dexamethasone-treated rats (up to 8 nmol of NO2- nmol of P450(-)1 min-1). A detailed study of the microsomal oxidation of some of these compounds was performed. Oxidation of N-(4-chlorophenyl)-N'-hydroxy-guanidine led to the formation of the corresponding urea and cyanamide in addition to NO, NO2-, and NO3-. Formation of all these products was dependent on NADPH, O2, and cytochromes P450. Oxidation of two arylamidoximes was found to occur with formation of the corresponding amides and nitriles in addition to nitrogen oxides. Oxidation of 4-(chlorophenyl)methyl ketone oxime gave the corresponding ketone and nitroalkane as well as NO, NO2-, and NO3-. These reactions were also dependent on cytochromes P450 and required NADPH and O2. Mechanistic experiments showed that microsomal oxidations of amidoximes to the corresponding nitriles and of ketoximes to the corresponding nitroalkanes are not inhibited by superoxide dismutase (SOD) and are performed by a cytochrome P450 active species, presumably the high-valent P450-iron-oxo complex. On the contrary, microsomal oxidation of N-hydroxyguanidines to the corresponding cyanamides was greatly inhibited by SOD and appeared to be mainly due to O2*- derived from the oxidase function of cytochromes P450. Similarly, microsomal oxidations of N-hydroxyguanidines and amidoximes to the corresponding ureas and amides were also found to be mainly performed by O2*-, as shown by the great inhibitory effect of SOD (70-100%) and the ability of the xanthine-xanthine oxidase system to give similar oxidation products. However, it is noteworthy that other species, such as the P450 Fe(II)-O2 complex, are also involved, to a minor extent, in the SOD-insensitive microsomal oxidative cleavages of compounds containing a C=N(OH) bond. Our results suggest a general mechanism for such oxidative cleavages of C=N(OH) bonds with formation of nitrogen oxides by cytochromes P450 and NO-synthases, with the involvement of O2*- and its Fe(III) complex [(FeIII-O2-) or (FeII-O2)] as main active species.
Jousserandot
A
A
Laboratoire de Chimie et Biochimie Pharmacologiques et Toxicologiques, URA 400 CNRS, Université Paris V, France.
Boucher
J L
JL
Henry
Y
Y
Niklaus
B
B
Clement
B
B
Mansuy
D
D
D000577:Amides / Q000378:metabolism*; D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases; D001550:Benzamidines / Q000494:pharmacology; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D003907:Dexamethasone / Q000008:administration & dosage; D004790:Enzyme Induction / Q000187:drug effects; D005609:Free Radicals / Q000037:antagonists & inhibitors / Q000378:metabolism; D006146:Guanidines / Q000378:metabolism*; D006860:Hydrogen Bonding / Q000187:drug effects; D006898:Hydroxylamines; D007274:Injections, Intraperitoneal; D008297:Male; D008748:Methylcholanthrene / Q000008:administration & dosage; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology*; D009589:Nitrogen Oxides / Q000378:metabolism*; D010084:Oxidation-Reduction / Q000187:drug effects; D010091:Oximes / Q000378:metabolism*; D010103:Oxygen Isotopes; D010634:Phenobarbital / Q000008:administration & dosage; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D039181:Steroid 16-alpha-Hydroxylase; D013250:Steroid Hydroxylases / Q000378:metabolism; D019820:Xanthine / Q000378:metabolism; D014969:Xanthine Oxidase / Q000378:metabolism
D000577:Amides; D001550:Benzamidines; D005609:Free Radicals; D006146:Guanidines; D006898:Hydroxylamines; D009589:Nitrogen Oxides; D010091:Oximes; D010103:Oxygen Isotopes; D019820:Xanthine; D008748:Methylcholanthrene; C053561:benzamidoxime; D003907:Dexamethasone; C029604:hydroxyguanidine; D003577:Cytochrome P-450 Enzyme System; D013250:Steroid Hydroxylases; D001189:Aryl Hydrocarbon Hydroxylases; D039181:Steroid 16-alpha-Hydroxylase; D014969:Xanthine Oxidase; D010634:Phenobarbital