Micropreparative isolation and NMR structure elucidation of metabolites of the drug candidate 1-isopropyl-4-(4-isopropylphenyl)-6-(prop-2-yn-1-yloxy) quinazolin-2(1H)-one from rat bile and urine

Blanz, Joachim; Délémonté, Thierry; Pearson, David; Luneau, Alexandre; Ritzau, Michael; Gertsch, Werner; Ramstein, Philippe; Dayer, Jérôme; Desrayaud, Sandrine; Braun, Elisabeth; Aichholz, Reiner

doi:10.1016/j.jchromb.2015.02.044

Cited by 7 publications

(5 citation statements)

References 19 publications

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“…2241. The study was performed as described previously . Briefly, the Sprague–Dawley male rat as specified in the general experimental part was used for this study.…”

Section: Methodsmentioning

confidence: 99%

“…The study was performed as described previously. [45] Briefly, the Sprague-Dawley male rat as specified in the general experimental part was used for this study. The day before drug administration, the animal was anaesthetized and then, under aseptic conditions, three catheters were successively implanted and fixed into its femoral artery, femoral vein, and bile duct for blood collection, drug administration, and continuous bile collection, respectively.…”

Section: In Vivo Experiments Using Bile Duct Cannulatedmentioning

confidence: 99%

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Design of A Metabolically Stable Tritium‐Tracer of the PI3Kδ‐Inhibitor CDZ173 (Leniolisib) as a Tool to Study Liver Metabolites

Bauer

Luu

Eggimann³

et al. 2018

Helvetica Chimica Acta

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In this disclosure, we summarize the preliminary metabolic profiling of the PI3Kδ inhibitor CDZ173 (leniolisib, 1a) obtained from incubations of the unlabeled compound and the synthesis of its metabolically stable tritium isotopologue 1b used for metabolite structure confirmation. Access to 1b was achieved when a halogenated precursor was subject to Hal/3H‐exchange. Hence, [3H]CDZ173 with specific activity 630 GBq/mmol, HPLC‐RA 97% and ee = 99.2% was obtained. Synthetic key to the precursor was using a bis‐halo‐pyridine in a Pd‐catalyzed mono‐amination of the tetrahydropyrido‐pyrimidine core. Stereochemistry of the synthetic precursors were confirmed by X‐ray analysis of the unlabeled bis‐halo‐pyridines and chiral HPLC of the tritiated material. The correct position of tritium label in the target, was confirmed by 3H‐NMR difference spectroscopy. Besides, we report on the validation of the radiotracer as a tool for pre‐clinical ADME in incubations with hepatocytes. Based on this data, we present a quantitative metabolite profile of leniolisib which was confirmed by independently synthesized metabolite references. The conformation of CDZ173 was investigated by NMR suggesting two different amide backbones each with specific pyrrolidine puckerings.

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“…2241. The study was performed as described previously . Briefly, the Sprague–Dawley male rat as specified in the general experimental part was used for this study.…”

Section: Methodsmentioning

confidence: 99%

Section: In Vivo Experiments Using Bile Duct Cannulatedmentioning

confidence: 99%

Design of A Metabolically Stable Tritium‐Tracer of the PI3Kδ‐Inhibitor CDZ173 (Leniolisib) as a Tool to Study Liver Metabolites

Bauer

Luu

Eggimann³

et al. 2018

Helvetica Chimica Acta

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“…The synthesis of the metabolite reference standards was based upon proposed chemical structures derived from product ion spectra or from NMR structural information obtained following micro-preparative isolation. [16] Several metabolites were also synthesized based upon predicted or anticipated formation. For example, a number of potential amide hydrolysis and N/O dealkylation products were tested.…”

Section: Metabolite Synthesismentioning

confidence: 99%

Evaluation of relative MS response factors of drug metabolites for semi‐quantitative assessment of chemical liabilities in drug discovery

et al. 2017

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Drug metabolism studies are performed in drug discovery to identify metabolic soft spots, detect potentially toxic or reactive metabolites and provide an early insight into potential species differences. The relative peak area approach is often used to semi-quantitatively estimate the abundance of metabolites. Differences in the liquid chromatography-mass spectrometry responses result in an underestimation or overestimation of the metabolite and misinterpretation of results. The relative MS response factors (RF) of 132 structurally diverse drug candidates and their 233 corresponding metabolites were evaluated using a capillary-liquid chromatography/high-resolution mass spectrometry system. All of the synthesized metabolites discussed here were previously identified as key biotransformation products in discovery investigations or predicted to be formed. The most commonly occurring biotransformation mechanisms such as oxygenation, dealkylation and amide cleavage are represented within this dataset. However, relatively few phase II metabolites were evaluated because of the limited availability of authentic standards. Approximately 85% of these metabolites had a relative RF in the range between 0.2 (fivefold under-prediction) and 2.0 (twofold over-prediction), and the median MS RF was 0.6. Exceptions to this included very small metabolites that were hardly detectable. Additional experiments performed to understand the impact of the MS platform, flow rate and concentration suggested that these parameters do not have a significant impact on the RF of the compounds tested. This indicates that the use of relative peak areas to semi-quantitatively estimate the abundance of metabolites is justified in the drug discovery setting in order to guide medicinal chemistry efforts. Copyright © 2017 John Wiley & Sons, Ltd.

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“…Previous studies have shown that euphorbias teroid is one of the main toxic and active components of Euphorbiae semen, and has also been used as a quantitative index for quality control in Chinese pharmacopoeia [6] However, there is still a lack of research on the metabolism of diterpenoids in euphorbi asteroid, which brings difficulties to the clinically safe use of this toxic Chinese medicine Thus, in order to further understand the toxic and active mechanisms of euphorbiaster oid, a systematic study of its metabolic profiles is of great significance. The in-depth study of drug metabolism can clarify the metabolic pathways as wel as the toxic and active mechanisms of drugs so as to ensure their safety and lay a theo retical foundation for clinical application and toxic and side effect detection [7]. There fore, it is of great significance and value to establish a sensitive and reliable analytica method for the identification of drug metabolites.…”

Section: Introductionmentioning

confidence: 99%

“…The advantages of the combination o UPLC-Q/TOF-MS and NMR have been verified in several reports in recent years [12][13][14] However, the sensitivity of NMR is not as good as that of mass spectrometry, and the presence of impurities in the sample has great influence on NMR signal. Therefore, it i The in-depth study of drug metabolism can clarify the metabolic pathways as well as the toxic and active mechanisms of drugs so as to ensure their safety and lay a theoretical foundation for clinical application and toxic and side effect detection [7]. Therefore, it is of great significance and value to establish a sensitive and reliable analytical method for the identification of drug metabolites.…”

Section: Introductionmentioning

confidence: 99%

Comprehensive Metabolic Profiling of Euphorbiasteroid in Rats by Integrating UPLC-Q/TOF-MS and NMR as Well as Microbial Biotransformation

Xiao¹,

Wei

Xin

et al. 2022

Metabolites

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Euphorbiasteroid, a lathyrane-type diterpene from Euphorbiae semen (the seeds of Euphorbia lathyris L.), has been shown to have a variety of pharmacological effects such as anti-tumor and anti-obesity. This study aims to investigate the metabolic profiles of euphorbiasteroid in rats and rat liver microsomes (RLMs) and Cunninghamella elegans bio-110930 by integrating ultra-performance liquid chromatography-quadrupole time-of-flight-mass spectrometry (UPLC-Q/TOF-MS), UNIFI software, and NMR techniques. A total of 31 metabolites were identified in rats. Twelve metabolites (M1–M5, M8, M12–M13, M16, M24–M25, and M29) were matched to the metabolites obtained by RLMs incubation and the microbial transformation of C. elegans bio-110930 and their structures were exactly determined through analysis of NMR spectroscopic data. In addition, the metabolic pathways of euphorbiasteroid were then clarified, mainly including hydroxylation, hydrolysis, oxygenation, sulfonation, and glycosylation. Finally, three metabolites, M3 (20-hydroxyl euphorbiasteroid), M24 (epoxylathyrol) and M25 (15-deacetyl euphorbiasteroid), showed significant cytotoxicity against four human cell lines with IC50 values from 3.60 μM to 40.74 μM. This is the first systematic investigation into the in vivo metabolic pathways of euphorbiasteroid and the cytotoxicity of its metabolites, which will be beneficial for better predicting the metabolism profile of euphorbiasteroid in humans and understanding its possible toxic material basis.

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Micropreparative isolation and NMR structure elucidation of metabolites of the drug candidate 1-isopropyl-4-(4-isopropylphenyl)-6-(prop-2-yn-1-yloxy) quinazolin-2(1H)-one from rat bile and urine

Cited by 7 publications

References 19 publications

Design of A Metabolically Stable Tritium‐Tracer of the PI3Kδ‐Inhibitor CDZ173 (Leniolisib) as a Tool to Study Liver Metabolites

Design of A Metabolically Stable Tritium‐Tracer of the PI3Kδ‐Inhibitor CDZ173 (Leniolisib) as a Tool to Study Liver Metabolites

Evaluation of relative MS response factors of drug metabolites for semi‐quantitative assessment of chemical liabilities in drug discovery

Comprehensive Metabolic Profiling of Euphorbiasteroid in Rats by Integrating UPLC-Q/TOF-MS and NMR as Well as Microbial Biotransformation

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