Glutathione alkylester isotopologue and method for detecting reactive metabolite
View Patent ↗[Problem] To provide a novel isotope-labeled compound that can be used as a trapping agent and that is useful for picking out drug-candidate compounds that produce reactive metabolites. [Solution] Provided is a glutathione alkylester isotopologue represented by general formula (1). In formula (1), R 1 represents a linear or branched alkoxy group in which at least one of carbon, oxygen, and hydrogen atoms contained therein is isotope-labeled and which has 1 to 8 carbon atoms or a cycloalkoxy group in which at least one of carbon, oxygen, and hydrogen atoms contained therein is isotope-labeled and which has 3 to 8 carbon atoms.
1. A glutathione alkyl ester isotopologue represented by general formula 3:
wherein D represents deuterium ( 2 H).
2. A method for detecting a reactive metabolite using the glutathione alkyl ester isotopologue according to claim 1 , the method comprising:
incubating a reaction sample containing the glutathione alkyl ester isotopologue,
an auxiliary detection compound which is a compound obtained by substituting at least one atom contained in the glutathione alkyl ester isotopologue with an atom having a mass number different from a mass number of the at least one atom, and a drug candidate compound in the presence of a drug metabolizing enzyme thereby forming a product containing a glutathione alkyl ester isotopologue-reactive metabolite adduct and an auxiliary detection compound-reactive metabolite adduct; and
detecting a mass peak of the formed glutathione alkyl ester isotopologue-reactive metabolite adduct and a mass peak of the formed auxiliary detection compound-reactive metabolite adduct through analysis using a liquid chromatography-mass spectrometer (LC-MS).
3. The method for detecting a reactive metabolite according to claim 2 , wherein a molar ratio of the glutathione alkyl ester isotopologue to the auxiliary detection compound in the reaction sample is 2:1 to 1:2.
4. The method for detecting a reactive metabolite according to claim 2 , the method further comprising:
adding dithiothreitol, 2-mercaptoethanol, or tris(2-carboxyethyl)phosphine to the product obtained by incubation; and
then performing the analysis using the liquid chromatography-mass spectrometer (LC-MS).
5. The method for detecting a reactive metabolite according to claim 2 , wherein a neutral loss scan method or a full scan method is performed in the analysis using the liquid chromatography-mass spectrometer (LC-MS).
6. The method for detecting a reactive metabolite according to claim 2 , wherein the auxiliary detection compound is a non-labeled compound of the glutathione alkyl ester isotopologue contained in the reaction sample.
7. A method for producing the glutathione alkyl ester isotopologue according to claim 1 , comprising reacting glutathione with ethanol-d6.