In vitro metabolism of MK-0767 [(+/-)-5-[(2,4-dioxothiazolidin-5-yl)methyl]-2-methoxy-N-[[(4-trifluoromethyl)-phenyl] methyl]benzamide], a peroxisome proliferator-activated receptor alpha/gamma agonist. II. Identification of metabolites by liquid chromatography-tandem mass spectrometry.
The in vitro metabolism of MK-0767 [(+/-)-5-[(2,4-dioxothiazolidin-5-yl) methyl]-2-methoxy-N-[[(4-trifluoromethyl)-phenyl] methyl]benzamide], a novel 2,4-thiazolidinedione (TZD)-containing peroxisome proliferator-activated receptor alpha/gamma agonist, was studied in rat, dog, monkey, and human liver microsomes and hepatocytes, as well as in recombinant human CYP3A4-containing microsomes. Twenty-two metabolites (some at trace levels) were detected by liquid chromatography-tandem mass spectrometry analysis. All appeared to be phase I metabolites except for a glucuronide conjugate of a hydroxylated metabolite that was detected at trace levels. A constant neutral loss scan experiment performed on a triple quadrupole mass spectrometer proved to be very useful for resolving the metabolites from endogenous compounds. It was observed that the initial site of metabolism of MK-0767 was at the TZD ring leading to two major metabolites, namely the 5-hydroxy-TZD metabolite (M24) and the mercapto metabolite (M22). The latter was formed via the cleavage of the TZD ring with the elimination of the carbonyl adjacent to the sulfur atom. The structure of M24 was established by accurate mass measurements and NMR analysis. This hydroxy-TZD metabolite might represent an important precursor for a group of metabolites formed by TZD ring opening and subsequent loss of the sulfur moiety. The mercapto metabolite, on the other hand, is probably the key precursor for the TZD ring-opened metabolites with retention of the sulfur, even though the detailed mechanism of the ring scission remains to be characterized. From these studies, it was concluded that the TZD ring was the major site of metabolism of MK-0767. All the metabolites produced in vitro from human preparations were detected in the corresponding preparations from the nonclinical species.
Merck Research Labora tories, Department of Drug Metabolism, RY80L-109, P.O. Box 2000, Rahway, NJ 07065, USA.
David Q
DQ
Liu
Bindhu V
BV
Karanam
George A
GA
Doss
Rick R
RR
Sidler
Stella H
SH
Vincent
Cornelis E C A
CE
Hop
D000818:Animals; D002250:Carbon Radioisotopes; D002853:Chromatography, Liquid / Q000379:methods*; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism; D004285:Dogs; D004353:Drug Evaluation, Preclinical / Q000379:methods; D020719:Glucuronides / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D000882:Haplorhini; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology; D009249:NADP / Q000378:metabolism; D047493:PPAR alpha / Q000737:chemistry / Q000378:metabolism*; D047495:PPAR gamma / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D012436:S-Adenosylmethionine / Q000378:metabolism; D013438:Sulfhydryl Compounds / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D013844:Thiazoles / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D045162:Thiazolidinediones / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology
D002250:Carbon Radioisotopes; D020719:Glucuronides; C487303:MK0767; D047493:PPAR alpha; D047495:PPAR gamma; D013438:Sulfhydryl Compounds; D013844:Thiazoles; D045162:Thiazolidinediones; D009249:NADP; D012436:S-Adenosylmethionine; D003577:Cytochrome P-450 Enzyme System; C104464:CYP3A protein, human; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human