Title:In vitro Drug Metabolism Investigation of 7-Ethoxycoumarin in Human, Monkey, Dog and Rat Hepatocytes by High Resolution LC-MS/MS
VOLUME: 12 ISSUE: 1
Author(s):Wan-Yong Feng*, Jenny Wen and Kathe Stauber
Affiliation:Drug Metabolism and Pharmacokinetics, Dart NeuroScience, San Diego, CA 92131, Drug Metabolism and Pharmacokinetics, Dart NeuroScience, San Diego, CA 92131, Drug Metabolism and Pharmacokinetics, Dart NeuroScience, San Diego, CA 92131
Keywords:7-ethoxycoumarin, metabolism, humans, monkeys, dogs, rats, hepatocytes.
Abstract:Background: Recently, it has been an increasing concern on the bioactivation and adverse
reactions associated with consumption of herbal and nature products. 7-Ethoxycoumarin is one of coumarin
family compounds, but little information is available regarding its potential reactive metabolites.
Method: 7-ethoxylcoumarin was incubated individually with human, monkey, dog and rat hepatocytes
for 2 hr, metabolites were detected, identified and characterized using high resolution liquid chromagraphy
– tandem mass spectrometry.
Results: Twenty-eight metabolites (M1 - M28) were detected and identified. O-deethylation, glucuronidation,
sulfation, oxygenation, oxidative ring-opening, hydrogenation, glutathionation, dehydrogenation,
cysteination, glucosidation, methylation, and hydrolysis were observed. At least sixteen metabolites
not reported previously, were newly identified. M1 (O-deethylation, mono-oxygenation and glucuronidation),
M3 (O-deethylation and glucuronidation), M5 (hydrolysis and mono-oxygenation), M14
(O-deethylation), M16 (hydrolysis), M22 (oxidative ring-opening and oxygenation) and M27 (monooxygenation)
exhibited high mass spectrometric responses in human hepatocytes. M3, M5, M8, M13
(mono-oxygenation), M14, M16, M18 (O-deethylation and sulfation), M22 and M27 exhibited high
mass spectrometric responses in monkey hepatocytes. M14, M16, M18, M20 (glutathionation and dehydrogenation)
and M27 exhibited high mass spectrometric responses in dog hepatocytes. M1 (Odeethylation,
mono-oxygenation and glucuronidation), M3, M5, M13, M14, M16, M17 (cysteination),
M18, M20, and M22 exhibited high mass spectrometric responses in rat hepatocytes.
Conclusion: Most of new metabolites via oxidative ring-opening and glutathionation were identified.
Species differences in metabolism of 7-ethoxylcoumarin in hepatocytes were observed. The analysis of
metabolites suggests that 7-ethoxylcoumarin may undergo 3,4-epoxidation responsible for formation of
glutathione and its derived cysteine conjugates, carboxylic acid and its glucuronides, glucosides and
sulfate.
