Analytical Tools and Approaches for Metabolite Identification in Early Drug Discovery

Chen, Yuan; Monshouwer, Mario; Fitch, William L.

doi:10.1007/s11095-006-9162-7

Cited by 42 publications

(31 citation statements)

References 94 publications

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“…Considering the typical concentration range (10 -50 M) in metabolite identification experiments (Chen et al, 2007), the upper limit concentration, that is, 50 M, was adopted. Furthermore, minimal sample cleanup was performed because the nature, number, and concentrations of metabolites present were unknown; therefore, it was impossible to determine whether any would be lost during sample preparation (Clarke et al, 2001).…”

Section: Discussionmentioning

confidence: 99%

“…However, there are a few severe draw-backs with regard to its experimental design. The substrate concentration adopted in that study far exceeded the upper limit of the typical concentration range (10 -50 M) in metabolite identification experiments (Chen et al, 2007). In addition, liquid-liquid extraction performed in that study was likely to cause the metabolites loss (Clarke et al, 2001) because physicochemical properties of metabolites present are unknown in a certain extraction solvent or a mixed extraction solvent system.…”

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confidence: 93%

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Identification of Stereoisomeric Metabolites of Meisoindigo in Rat Liver Microsomes by Achiral and Chiral Liquid Chromatography/Tandem Mass Spectrometry

Huang¹,

Goh²,

Ho³

2008

Drug Metab Dispos

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ABSTRACT:N-Methylisoindigotin, abbreviated as meisoindigo, has been a routine therapeutic agent in the clinical treatment of chronic myelogenous leukemia in China since the 1980s. However, information relevant to in vitro metabolism of meisoindigo is limited. In this study, in vitro stereoisomeric metabolites of meisoindigo in rat liver microsomes were identified for the first time by achiral and chiral liquid chromatography/tandem mass spectrometry, together with proton NMR spectroscopy and synchrotron infrared spectroscopy. The major in vitro phase I metabolites of meisoindigo were tentatively identified as stereoselective-reduced meisoindigo, which comprised a pair of (3-R, 3-R) and (3-S, 3-S) enantiomers with lower abundance, as well as another pair of (3-R, 3-S) and (3-S, 3-R) enantiomers with higher abundance. One type of minor in vitro metabolites was tentatively identified as stereoselective N-demethyl-reduced meisoindigo including a pair of (3-R, 3-R) and (3-S, 3-S) enantiomers, as well as one meso compound. Another type of minor in vitro metabolites was tentatively identified as both stereoselective and regioselective monohydroxyl-reduced meisoindigo. Based on the metabolite profiling, three parallel metabolic pathways of meisoindigo in rat liver microsomes were proposed.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 93%

Identification of Stereoisomeric Metabolites of Meisoindigo in Rat Liver Microsomes by Achiral and Chiral Liquid Chromatography/Tandem Mass Spectrometry

Huang¹,

Goh²,

Ho³

2008

Drug Metab Dispos

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“…A microbial system also provides enough putative metabolites under milder conditions than those required by chemical systems. Thus, fermentation is used to scale up the synthesis of metabolites for structural confirmation by nuclear magnetic resonance (NMR) [8] .…”

Section: Introductionmentioning

confidence: 99%

Biotransformation of metoprolol by the fungus Cunninghamella blakesleeana

Huang

Chen

et al. 2007

Acta Pharmacologica Sinica

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Aim: To investigate the biotransformation of metoprolol, a β1‐cardioselective adrenoceptor antagonist, by filamentous fungus, and to compare the parallels between microbial transformation and mammalian metabolism. Methods: Five strains of Cunninghamella (Celegans AS 3.156, Celegans AS 3.2028, C echinulata AS 3.2004, C blakesleeana AS 3.153 and AS 3.910) were screened for the ability to transform metoprolol. The metabolites of metoprolol produced by C blakesleeana AS 3.153 were separated and assayed by liquid chromatography‐tandem mass spectrometry (LC/MSn). The major metabolites were isolated by semipreparative HPLC and the structures were identified by a combination of LC/MSn and nuclear magnetic resonance analysis. Results: Metoprolol was transformed to 7 metabolites; 2 were identified as new metabolites and 5 were known metabolites in mammals. Conclusion: The microbial transformation of metoprolol was similar to the metabolism in mammals. The fungi belonging to Cunninghamella species could be used as complementary models for predicting in vivo metabolism and producing quantities of metabolite references for drugs like metoprolol.

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“…Currently, liquid chromatography (LC) coupled with electrospray ionization (ESI)-MS has been widely used to detect and identify trace levels of drugs and their metabolites in various biological samples due to its high sensitivity and selectivity. [17][18][19][20] Conventional LC, however, is often a slow technique for analyzing complex biological samples, 15,16 and the commonly used quadrupole or ion trap mass analyzers provide limited mass resolution especially for complex sample analyses. Therefore, development of new analytical techniques to overcome these limitations is highly desirable.…”

mentioning

confidence: 99%

Identification of metabolites of Danggui Buxue Tang in rat urine by liquid chromatography coupled with electrospray ionization time‐of‐flight mass spectrometry

et al. 2009

Rapid Comm Mass Spectrometry

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A method coupling liquid chromatography with electrospray ionization time-of-flight mass spectrometry (LC/ESI-TOF/MS) has been developed for rapid and sensitive analysis of rat urinary metabolite profile of Danggui Buxue Tang (DBT), a well-known Chinese herbal formula. After oral administration of DBT, urine samples were collected during 0-24 h, and then pretreated by solid-phase extraction. A total of 68 compounds including 13 parent compounds and 55 metabolites were detected in the drug-containing urines compared with blank urines. The total analytical time was less than 20 min. Metabolites of DBT were identified using dynamic adjustment of the fragmentor voltage to produce structure-relevant fragment ions. By using this approach, the mass accuracy of precursor and fragment ions was typically within +/-5 ppm of the theoretical values, and enabled the identification of 43 metabolites including 27 isoflavanoid and 16 phthalide metabolites. Our results indicated that glucuronidation and sulfation were the major metabolic pathways of isoflavonoids, while glutathione conjugation, glucuronidation and sulfation were the main metabolic pathways of phthalides. No saponin-related metabolites were detected. The results of the present study provided important structural information relating to the metabolism of DBT. Furthermore, this work demonstrated the potential of the LC/ESI-TOF/MS approach for identification of metabolites from Chinese herbal medicines in urine.

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Analytical Tools and Approaches for Metabolite Identification in Early Drug Discovery

Cited by 42 publications

References 94 publications

Identification of Stereoisomeric Metabolites of Meisoindigo in Rat Liver Microsomes by Achiral and Chiral Liquid Chromatography/Tandem Mass Spectrometry

Identification of Stereoisomeric Metabolites of Meisoindigo in Rat Liver Microsomes by Achiral and Chiral Liquid Chromatography/Tandem Mass Spectrometry

Biotransformation of metoprolol by the fungus Cunninghamella blakesleeana

Identification of metabolites of Danggui Buxue Tang in rat urine by liquid chromatography coupled with electrospray ionization time‐of‐flight mass spectrometry

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