Carboxylesterase catalyzed 18O-labeling of carboxylic acid and its potential application in LC-MS/MS based quantification of drug metabolites

Yu, Zaikuan; Roesner, Joseph M.; Lutz, Ryan; Liang, Yuexia; Baker, James Α.; Smith, Dustin M.; Fan, Peter W.

doi:10.1016/j.dmpk.2019.05.004

Cited by 7 publications

(5 citation statements)

References 22 publications

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“…Although 18 O-labeling from H 2 18 O has been used extensively in quantitative proteomics, its application in small molecule analysis remains largely underexplored. We previously reported 18 O-labeling of carboxylic acid-containing compounds through the use of carboxylesterases . In the present study, we expand the scope of 18 O-labeling to AGs to differentiate between migrated and unmigrated species.…”

Section: Introductionmentioning

confidence: 91%

¹⁸O-Enabled High-Throughput Acyl Glucuronide Stability Assay

Lê

Tang

et al. 2022

Chem. Res. Toxicol.

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Acyl glucuronides (AGs) are common metabolites of carboxylic acid-containing compounds. In some circumstances, AGs are suspected to be involved in drug toxicity due to formation of acyl migration products that bind covalently to cellular components. The risk of this adverse effect has been found to be correlated with the chemical stability of the AG, and assays have been described that monitor acyl migration by liquid chromatography coupled with mass spectrometry (LC-MS). This analysis can be challenging as it requires baseline chromatographic separation of the unmigrated 1-β-acyl glucuronide from the migrated isomers and thus needs to be individually optimized for each aglycone. Therefore, a high-throughput assay that eliminates LC method development is desirable. Herein, we report an improved acyl glucuronide stability assay based on the rate of 18O-incorporation from [18O] water, which is compatible with high-throughput bioanalytical LC-MS workflows. Synthetic AGs with shorter migration half-lives showed faster incorporation of 18O. The level of differential incorporation of 18O following a 24 h incubation correlates well with the migration tendency of AGs. This assay was developed further, exploring in situ generation of AGs by human hepatic microsomal fraction. The results from 18 in situ-formed acyl glucuronides were similar to those obtained using authentic reference standards. In this format, this new 18O-labeling method offers a simplified workflow, requires no LC method development or AG reference standard, and thus facilitates AG liability assessment in early drug discovery.

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Section: Introductionmentioning

confidence: 91%

¹⁸O-Enabled High-Throughput Acyl Glucuronide Stability Assay

Lê

Tang

et al. 2022

Chem. Res. Toxicol.

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“…The carboxylic group is one such ubiquitous moiety with valuable intrinsic chemical abilities that contributes to mammalian physiology and endogenous biochemical processes . Carboxylic acid ionizes and gives better solubility at pH 7.4 making it a suitable moiety for drug development as it can be easily metabolized (normal pH of blood 7.35–7.45) . Other properties such as hydrogen bonding and p K a play a crucial role in the ligand-protein binding depending on inductive effect, neighboring groups, chain length, and so forth. , In literature, more than 450 drugs are enlisted containing carboxylic group on aromatic (Diclofenac, ibuprofen, aspirin, and so forth), heterocyclic rings (Ciprofloxacin, Amoxicillin, Norfloxacin and Levofloxacin), statins (Epanova, Atorvastatin, Fluvastatin, and so forth), β-lactam antibiotics (Penicillin, Cephalosporins, Cephamycins, and so forth), fibrates (Gemfibrozil), NSAIDs (Indomethacin, Naproxen, and so forth), and food preservatives …”

Section: Introductionmentioning

confidence: 99%

Highly Efficient Electrocarboxylation Method to Synthesize Novel Acid Derivatives of 1,4-Dihydropyridines and to Study Their Antimicrobial Activity

et al. 2022

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1,4-Dihydropyridines (1,4-DHPs) hold a top-notch position in the pharmaceutical world due to a broader spectrum of applications, whereas the carboxylic moiety has been an integral part of the physiological world, effective food preservatives, and antimicrobial agents. Seeking the enormous potential and applications of these two classes, we worked to combine these to synthesize 2,2′-[3,5-bis(ethoxycarbonyl)-4-phenyl-1,4-dihydropyridine-2,6-diyl]diacetic acid the novel dicarboxylic derivatives of 1,4-DHP ( 9a – k ) achieved via the electro-carboxylation of tetrasubstituted-1,4-dihydropyridines ( 8a – k ) derivatives using Mg–Pt electrodes in an undivided cell. The targeted compounds were established by 1 H, 13 C NMR, IR, and ESI-MS. Further, the synthesized compounds show excellent resistance against various microbes and the activity increased 2–3 folds after the introduction of acid groups. Compound 9b (against E. coli , S. aureus , B. subtilis , A. niger , and P. glabrum ), 9d (against E. coli , K. pneumonia , S. aureus , A. janus , and F. oxysporum) , 9f (against E. coli and P. fluorescens ), and 9k (against F. oxysporum and P. glabrum) were found to be highly active at 4 μg/mL with reference to standard amoxicillin and fluconazole. Further, the present synthetic protocol would open new gates for other researchers to develop new molecules by bioisosteres of these substrates.

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“…In-source oxygen isotope exchange was reported only for inorganic molecules . Proteolytic 18 O incorporation to peptides was demonstrated and became a fundament for producing standards for quantitative proteomics or to investigate metabolic pathways. , For such purposes, the reaction was often conducted with enzymatic systems that facilitated 18 O incorporation . For labeled data processing purposes, isoMetlin database and geoRge software have been developed.…”

Section: Introductionmentioning

confidence: 99%

Oxygen Isotope Exchange Reaction for Untargeted LC–MS Analysis

Osipenko

Zherebker

Rumiantseva

et al. 2022

J. Am. Soc. Mass Spectrom.

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LC–MS is a key technique for the identification of small molecules in complex samples. Accurate mass, retention time, and fragmentation spectra from LC–MS experiments are compared to reference values for pure chemical standards. However, this information is often unavailable or insufficient, leading to an assignment to a list of candidates instead of a single hit; therefore, additional features are desired to filter candidates. One such promising feature is the number of specific functional groups of a molecule that can be counted via derivatization or isotope-exchange techniques. Hydrogen/deuterium exchange (HDX) is the most widespread implementation of isotope exchange for mass spectrometry, while oxygen 16O/18O exchange is not applied as frequently as HDX. Nevertheless, it is known that some functional groups may be selectively exchanged in 18O enriched media. Here, we propose an implementation of 16O/18O isotope exchange to highlight various functional groups. We evaluated the possibility of using the number of exchanged oxygen atoms as a descriptor to filter database candidates in untargeted LC–MS-based workflows. It was shown that 16O/18O exchange provides 62% (median, n = 45) search space reduction for a panel of drug molecules. Additionally, it was demonstrated that studying the fragmentation spectra after 16O/18O can aid in eliminating false positives and, in some cases, help to annotate fragments formed with water traces in the collisional cell.

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Carboxylesterase catalyzed 18O-labeling of carboxylic acid and its potential application in LC-MS/MS based quantification of drug metabolites

Cited by 7 publications

References 22 publications

¹⁸O-Enabled High-Throughput Acyl Glucuronide Stability Assay

¹⁸O-Enabled High-Throughput Acyl Glucuronide Stability Assay

Highly Efficient Electrocarboxylation Method to Synthesize Novel Acid Derivatives of 1,4-Dihydropyridines and to Study Their Antimicrobial Activity

Oxygen Isotope Exchange Reaction for Untargeted LC–MS Analysis

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Carboxylesterase catalyzed 18O-labeling of carboxylic acid and its potential application in LC-MS/MS based quantification of drug metabolites

Cited by 7 publications

References 22 publications

18O-Enabled High-Throughput Acyl Glucuronide Stability Assay

18O-Enabled High-Throughput Acyl Glucuronide Stability Assay

Highly Efficient Electrocarboxylation Method to Synthesize Novel Acid Derivatives of 1,4-Dihydropyridines and to Study Their Antimicrobial Activity

Oxygen Isotope Exchange Reaction for Untargeted LC–MS Analysis

Contact Info

Product

Resources

About

¹⁸O-Enabled High-Throughput Acyl Glucuronide Stability Assay

¹⁸O-Enabled High-Throughput Acyl Glucuronide Stability Assay