Development of an LC–MS/MS assay for the quantitative determination of the intracellular 5-fluorouracil nucleotides responsible for the anticancer effect of 5-fluorouracil

Derissen, Ellen J. B.; Hillebrand, M. J. X.; Rosing, Hilde; Schellens, Jan H.M.; Beijnen, Jos H.

doi:10.1016/j.jpba.2015.02.051

Cited by 26 publications

(26 citation statements)

References 23 publications

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“…This would be the case if the transporters involved in the cellular uptake or the enzymes involved in the intracellular ribosylation or phosphorylation would become saturated above a certain 5-FU concentration. Remarkably, the FUTP concentrations which we reported in our analytical paper, measured 30 min after an intravenous 5-FU bolus (400 mg m À2 in 30 min) were in the same range as the FUTP concentrations measured after 14 days of capecitabine treatment twice daily (4.7-11 μM vs. 0.64-14 μM) [13]. This indicates that also brief, high 5-FU exposure can rapidly lead to high intracellular FUTP concentrations and the capacities of the cell membrane transporters and intracellular enzymes are probably not the limiting factors.…”

Section: Figuresupporting

confidence: 82%

“…The LLQs in PBMC lysate were 0.488 nM for FUTP, 1.66 nM for FdUTP and 0.748 nM for FdUMP. Accuracies were between −2.2 and 7.0% deviation for all analytes, and the coefficient of variation values were ≤4.9% .…”

Section: Methodsmentioning

confidence: 90%

“…The aim of the current study was to explore the intracellular pharmacokinetics (PK) of the three pharmacologically active 5‐FU nucleotides during capecitabine treatment. Except for a small pilot in our previous publication on the development of a bioanalytical assay , this is, to our knowledge, the first time that intracellular 5‐FU nucleotides were quantified during capecitabine treatment.…”

Section: Introductionmentioning

confidence: 95%

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Exploring the intracellular pharmacokinetics of the 5‐fluorouracil nucleotides during capecitabine treatment

Derissen

Jacobs

Huitema

et al. 2016

Brit J Clinical Pharma

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Keywords 5-fluorouracil, 5-fluorouridine triphosphate (FUTP), capecitabine, nucleotides, peripheral blood mononuclear cells (PBMCs), pharmacokinetics AIM Three intracellularly formed metabolites are responsible for the antineoplastic effect of capecitabine: 5-fluorouridine 5′-triphosphate (FUTP), 5-fluoro-2′-deoxyuridine 5′-triphosphate (FdUTP), and 5-fluoro-2′-deoxyuridine 5′-monophosphate (FdUMP). The objective of this study was to explore the pharmacokinetics of these intracellular metabolites during capecitabine treatment. METHODSSerial plasma and peripheral blood mononuclear cell (PBMC) samples were collected from 13 patients treated with capecitabine 1000 mg QD (group A) and eight patients receiving capecitabine 850 mg m À2 BID for fourteen days, every three weeks (group B). Samples were collected on day 1 and, for four patients of group B, also on day 14. The capecitabine and 5-fluorouracil (5-FU) plasma concentrations and intracellular metabolite concentrations were determined using LC-MS/MS. Pharmacokinetic parameters were estimated using non-compartmental analysis. RESULTSOnly FUTP could be measured in the PBMC samples. The FdUTP and FdUMP concentrations were below the detection limits (LOD). No significant correlation was found between the plasma 5-FU and intracellular FUTP exposure. The FUTP concentrationtime profiles demonstrated considerable inter-individual variation and accumulation of the metabolite in PBMCs. FUTP levels ranged between

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Section: Figuresupporting

confidence: 82%

Section: Methodsmentioning

confidence: 90%

Section: Introductionmentioning

confidence: 95%

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Exploring the intracellular pharmacokinetics of the 5‐fluorouracil nucleotides during capecitabine treatment

Derissen

Jacobs

Huitema

et al. 2016

Brit J Clinical Pharma

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“…the use of radioactive‐labeled nucleotides or lack of sensitivity) (Procházková, Liu, Friess, Aebi, & Thormann, ; Weckbecker, ). To date and to our knowledge only Derissen, Hillebrand, Rosing, Schellens, and Beijnen () have reported the development of an LC‐MS/MS assay that can quantify FdUMP, FUTP and FdUTP intracellularly, in a clinical setting (Derissen et al, ). Quantification of FdUMP, FUTP and FdUTP in peripheral blood mononuclear cells (PBMCs) by cell lysis and subsequent extraction with methanol was described.…”

Section: Intracellular Metabolites—fdump Futp and Fdutpmentioning

confidence: 99%

A review of the bioanalytical methods for the quantitative determination of capecitabine and its metabolites in biological matrices

Knikman

Rosing

Guchelaar

et al. 2020

Biomedical Chromatography

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The bioanalysis of the oral anticancer drug capecitabine and its metabolites has been investigated extensively over the past years. This paper reviews methods for the bioanalysis of capecitabine and its metabolites. The focus of this review will be on sample pre‐treatment, chromatography and detection. Furthermore, the choice of standards and analytical problems encountered during analysis of capecitabine and its metabolites in biological matrices will be discussed. The major challenges in the bioanalysis of capecitabine and its metabolites are the simultaneous extraction and analysis due to the differences in polarity of the analytes. Furthermore we evaluate currently described methods for the quantification of capecitabine and its metabolites. Future wishes and perspectives are stated that could serve as an inspiration for further development of assays for the quantification of capecitabine and its metabolites.

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“…Because of their hydrophilicity, triphosphate nucleosides have very poor retention when separated on traditional reverse-phase high-performance liquid chromatography columns, and thus another chromatographic approach is required. The possibility is using ion-pairing [12, 16, 19-21, 23, 26, 31, 32, 41, 47] or anion exchange chromatography [13,17,18,27,42]. These approaches enhance the retention on RP columns, but they are, however, compromised by the high levels of additives in mobile phases (ammonium salts and alkylamines), which cause significant ion suppression, poor robustness and instability of the ion exchange columns, and also source pollution, which remains a major problem [10,16,60,76].…”

mentioning

confidence: 99%

Mass Spectrometry for the Sensitive Analysis of Intracellular Nucleotides and Analogues

Mičová¹,

Friedecký²,

Adam³

2017

Mass Spectrometry

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Nowadays, mass spectrometry is very important and widely applied tool in nucleotide analysis. As a result of technological advances in sample preparation, separation and mass spectrometry detection, the developed methods allow sensitive and selective measurement of polar compounds occurring in low levels in various biological matrices. This enables more potential uses in clinical field. Direct methods require no special sample pre-treatment before analysis in contrast to indirect methods, where fractionation, dephosphorylation and purification are needed. The use of ion-pairing agent, ion exchange chromatography with pH gradient, porous graphitic carbon columns and HILIC in liquid chromatography represents the most common methods of nucleotide analysis. High separation efficiency is also achieved with the use of CE with MS detection. Analysis of nucleotides was also described by the means of MALDI-TOF, but poor reproducibility and lack of applications make a limitation for this approach. The chapter summarizes different techniques and approaches for determination of endogenous nucleotides and its analogues in various clinical applications. or deoxyribose) through a glycosidic bond. Nucleotides are formed by phosphorylation of nucleoside molecule. They represent a fundamental building block of the structure of nucleic acids formed by the polymerase-mediated synthesis of DNA and RNA from deoxynucleotides and nucleotides, respectively. All deoxynucleotides are synthesized from the corresponding nucleotides. Nucleotides play another important role in enzyme activation and metabolism. They are incorporated into important cofactors of enzymatic reactions such as transfer of various groups by coenzyme A (e.g. acetyl-CoA and succinyl-CoA) in many metabolic processes, NAD, NADP and FAD as coenzymes of oxidoreductases catalyses redox reactions in the cell. ATP and other nucleoside triphosphates are engaged as source energy providers. In the form of coenzymes, they transfer phosphate residues or nucleotide constituent and participate in the activation (e.g. uridine diphosphate glucose for pasting glucose units to polymeric saccharides or cytidine diphosphate-choline for phospholipid synthesis). Cyclic nucleotides act as secondary messengers in many signal transduction pathways. The most important nucleotide signal molecule is cyclic adenosine monophosphate (cAMP), which is an activator of protein kinases and participates in many metabolic pathways by transferring the effect of hormones into cells. Analogical molecule-cyclic guanosine monophosphate (cGMP)-also acts as a secondary messenger in the phototransduction process.Nucleotide metabolism is one of the main therapeutic cellular targets. Synthetic analogues of these compounds have been widely applied in anti-cancer, anti-viral and immunosuppressive therapies [1][2][3][4][5][6]. Nucleoside analogues (e.g. cytarabine, gemcitabine and zidovudine) are modified at the base or sugar moiety. This 'prodrug' form requires intracellular activation by phosphorylation to mono, di or...

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Development of an LC–MS/MS assay for the quantitative determination of the intracellular 5-fluorouracil nucleotides responsible for the anticancer effect of 5-fluorouracil

Cited by 26 publications

References 23 publications

Exploring the intracellular pharmacokinetics of the 5‐fluorouracil nucleotides during capecitabine treatment

Exploring the intracellular pharmacokinetics of the 5‐fluorouracil nucleotides during capecitabine treatment

A review of the bioanalytical methods for the quantitative determination of capecitabine and its metabolites in biological matrices

Mass Spectrometry for the Sensitive Analysis of Intracellular Nucleotides and Analogues

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