Identification of 20(S)-protopanaxadiol metabolites in human liver microsomes and human hepatocytes.
20(S)-Protopanaxadiol (PPD, 1) is one of the aglycones of the ginsenosides and has a wide range of pharmacological activities. At present, PPD has progressed to early clinical trials as an antidepressant. In this study, its fate in mixed human liver microsomes (HLMs) and human hepatocytes was examined for the first time. By using liquid chromatography-electrospray ionization ion trap mass spectrometry, 24 metabolites were found. Four metabolites were isolated, and their structures were elucidated as (20S,24S)-epoxydammarane-3,12,25-triol (2), (20S,24R)-epoxydammarane-3,12,25-triol (3), (20S,24S)-epoxydammarane-12,25-diol-3-one (4), and (20S,24R)-epoxydammarane-12,25-diol-3-one (5) based on a detailed analysis of their spectroscopic data. The predominant metabolic pathway of PPD observed was the oxidation of the 24,25-double bond to yield 24,25-epoxides, followed by hydrolysis and rearrangement to form the corresponding 24,25-vicinal diol derivatives (M6) and the 20,24-oxide form (2 and 3). Further sequential metabolites (M2-M5) were also detected through the hydroxylation and dehydrogenation of 2 and 3. All of the phase I metabolites except for M1-1 possess a hydroxyl group at C-25 of the side chain, which was newly formed by biotransformation. Two glucuronide conjugates (M7) attributed to 2 and 3 were detected in human hepatocyte incubations, and their conjugation sites were tentatively assigned to the 25-hydroxyl group. The findings of this study strongly suggested that the formation of the 25-hydroxyl group is very important for the elimination of PPD.
Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China.
Liang
L
Li
Xiaoyan
X
Chen
Dan
D
Li
Dafang
D
Zhong
D000818:Animals; D000928:Antidepressive Agents / Q000032:analysis / Q000737:chemistry / Q000378:metabolism* / Q000493:pharmacokinetics; D001711:Biotransformation; D002851:Chromatography, High Pressure Liquid; D004852:Epoxy Compounds / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D005243:Feces / Q000737:chemistry; D036145:Ginsenosides / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D020719:Glucuronides / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism*; D015394:Molecular Structure; D051381:Rats; D017207:Rats, Sprague-Dawley; D012502:Sapogenins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism* / Q000493:pharmacokinetics; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry
C000601989:20,24-epoxydammarane-3,12,25-triol; D000928:Antidepressive Agents; D004852:Epoxy Compounds; D036145:Ginsenosides; D020719:Glucuronides; D012502:Sapogenins; C062916:protopanaxadiol