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1.
Analyst ; 142(20): 3771-3796, 2017 Oct 09.
Article in English | MEDLINE | ID: mdl-28858342

ABSTRACT

While NMR is the most used analytical method for determining the molecular structure of isolated chemical entities, small compounds as well as macromolecules, its capability of analysing complex mixtures is less known. The advent of Diffusion Ordered SpectroscopY (DOSY) NMR has made diffusion experiments popular, enabling diffusion coefficients to be routinely measured and used to characterize chemical systems in solution. Indeed, since the translational diffusion coefficients of molecular species reflect their effective sizes and shapes, DOSY NMR allows the separation of the chemical entities present in multicomponent systems and, as in all diffusion NMR experiments, provides information on their intermolecular interactions as well as on their size and shape. The main aim of this review is to present an overview of the DOSY NMR mapping and its applications. The paper starts with a brief introduction to pulsed-field gradient (PFG) NMR and then focuses on the methodological procedures that can be used to perform good diffusion data acquisition and to obtain good-quality DOSY maps. The second part describes, through selected literature examples, different applications of DOSY NMR to demonstrate the potential of the method for (i) unravelling the components of complex matrices comprising pharmaceuticals, dietary supplements, foods and beverages, and biological extracts, and (ii) probing intermolecular interactions and evaluating association constants between different hosts and guests, as well as estimating the sizes and molecular weights of molecular species.

2.
J Microencapsul ; 34(2): 162-170, 2017 Mar.
Article in English | MEDLINE | ID: mdl-28345377

ABSTRACT

The association chitosan/linoleic acid/lactobionic acid in aqueous solution spontaneously led to the formation of stable microparticles with a liquid hydrophobic core consisting of linoleic acid surrounded by a shell of chitosan/lactobionic acid. The originality of the microparticles arises from the fact that they are formed by the association of three ingredients of cosmetic interest, including a skin penetration enhancer (linoleic acid). Dynamic light scattering (DLS) measurements showed microparticles with a mean diameter of 1-2 µm. The presence of a hydrophobic liquid core was observed by transmission electron microscopy (TEM). The ability of these microparticles to encapsulate phenylethyl resorcinol, a hydrophobic skin lightener, was evaluated and its encapsulation was confirmed thanks to T2 measurements and nuclear Overhauser effects (nOe) signs.


Subject(s)
Chitosan/chemistry , Cosmetics/chemistry , Disaccharides/chemistry , Linoleic Acid/chemistry , Particle Size
3.
J Pharm Biomed Anal ; 55(1): 1-15, 2011 Apr 28.
Article in English | MEDLINE | ID: mdl-21237608

ABSTRACT

This article focuses on the description of some of the NMR techniques used in the field of biomedical and pharmaceutical research. Indeed, the NMR method has special characteristics which make it uniquely suitable for these kinds of studies. It is non-selective so that all the low molecular weight compounds in the sample investigated are detected simultaneously in a single run. NMR also provides rich structural information which is an important asset to characterize complex mixture components. NMR is quantitative, i.e. the area of a NMR signal is directly proportional to the number of corresponding nuclei and thus, at variance with other techniques, the response factor is not dependent on the molecular structure. It is also a non-invasive tool that permits in vivo studies in humans. Compared with other techniques, NMR is significantly insensitive, which represents the main drawback of the technique. The recent technological developments of the technique have nevertheless considerably improved its sensitivity. The first part of this article presents an overview of the advantages and limitations of NMR for in vitro quantitative analysis of complex matrices in liquid or semi-solid phases. The second part deals with the NMR-based metabolomics methodology. The third part describes the in vivo clinical magnetic resonance spectroscopy techniques. The fourth part reports some examples of NMR applications in the biomedical and pharmaceutical research fields.


Subject(s)
Magnetic Resonance Spectroscopy/methods , Technology, Pharmaceutical , Animals , Biomedical Technology , Complex Mixtures/chemistry , Drug Contamination/prevention & control , Humans , Magnetic Resonance Imaging/methods , Metabolomics/methods , Neoplasm Staging/methods , Neoplasms/diagnosis , Neoplasms/metabolism , Neoplasms/pathology , Quality Control
4.
Anal Bioanal Chem ; 399(2): 987-99, 2011 Jan.
Article in English | MEDLINE | ID: mdl-21069302

ABSTRACT

The aim of this study was to define the optimal pH for (1)H nuclear magnetic resonance (NMR) spectroscopy analysis of perchloric acid or methanol-chloroform-water extracts from brain tumor cells and tissues. The systematic study of the proton chemical shift variations as a function of pH of 13 brain metabolites in model solutions demonstrated that recording (1)H NMR spectra at pH 10 allowed resolving resonances that are overlapped at pH 7, especially in the 3.2-3.3 ppm choline-containing-compounds region. (1)H NMR analysis of extracts at pH 7 or 10 showed that quantitative measurements of lactate, alanine, glutamate, glutamine (Gln), creatine + phosphocreatine and myo-inositol (m-Ino) can be readily performed at both pHs. The concentrations of glycerophosphocholine, phosphocholine and choline that are crucial metabolites for tumor brain malignancy grading were accurately measured at pH 10 only. Indeed, the resonances of their trimethylammonium moieties are cleared of any overlapping signal, especially those of taurine (Tau) and phosphoethanolamine. The four non-ionizable Tau protons resonating as a singlet in a non-congested spectral region permits an easier and more accurate quantitation of this apoptosis marker at pH 10 than at pH 7 where the triplet at 3.43 ppm can be overlapped with the signals of glucose or have an intensity too low to be measured. Glycine concentration was determined indirectly at both pHs after subtracting the contribution of the overlapped signals of m-Ino at pH 7 or Gln at pH 10.


Subject(s)
Brain Chemistry , Brain Neoplasms/chemistry , Choline/analysis , Glioma/chemistry , Magnetic Resonance Spectroscopy/methods , Taurine/analysis , Brain/metabolism , Brain Neoplasms/metabolism , Choline/metabolism , Glioma/metabolism , Humans , Hydrogen-Ion Concentration , Taurine/metabolism
5.
Anal Bioanal Chem ; 398(6): 2723-30, 2010 Nov.
Article in English | MEDLINE | ID: mdl-20865406

ABSTRACT

A new method based on hydrophilic interaction chromatography-electrospray ionisation-tandem mass spectrometry (HILIC-ESI-MS/MS) coupled to the use of a stable isotope labelled substrate was developed to study the metabolism of choline (Cho) compounds in two human glioblastoma multiform (GBM) cell lines with different responses to ionising radiation. Analysis was performed in the positive ion mode using multiple reaction monitoring. This fast, sensitive and selective method enabled the profiling of both hydrophilic and lipophilic Cho-containing compounds, to analyse specifically different phosphatidylcholine (PtdCho) molecular species, and to measure simultaneously native and labelled Cho metabolites. Radioresistant (SF763) and radiosensitive (SF767) cells were incubated for 8 h with d(9)-Cho. Higher native Cho and phosphocholine (PCho) concentrations and higher uptake of d(9)-Cho and formation of d(9)-PCho were found in the radioresistant cell line. The similar low concentrations of native cytidine 5'-diphosphocholine (CDP-Cho) and d(9)-CDP-Cho in both cell lines show that CDP-Cho is the limiting metabolite in the two models. The turnovers (percentage of each d(9)-Cho compound in its respective pool, i.e. native + labelled) were lower in radioresistant cells for all Cho compounds, suggesting a global PtdCho metabolism more active in radiosensitive cells that could be related to their higher proliferation rate.


Subject(s)
Brain Neoplasms/radiotherapy , Choline/analysis , Chromatography, Liquid/methods , Glioblastoma/radiotherapy , Spectrometry, Mass, Electrospray Ionization/methods , Tandem Mass Spectrometry/methods , Brain Neoplasms/metabolism , Cell Culture Techniques , Cell Line, Tumor/metabolism , Cell Line, Tumor/radiation effects , Choline/metabolism , Chromatography, Liquid/instrumentation , Glioblastoma/metabolism , Humans , Hydrophobic and Hydrophilic Interactions , Spectrometry, Mass, Electrospray Ionization/instrumentation , Tandem Mass Spectrometry/instrumentation
6.
Article in English | MEDLINE | ID: mdl-20437283

ABSTRACT

Twenty herbal medicines or dietary supplements marketed as natural slimming products were analysed by diffusion ordered spectroscopy (DOSY) 1H-nuclear magnetic resonance (NMR) and DOSY-COSY 1H-NMR. The method allows analysis of the whole sample with the detection of both active and inactive ingredients in these complex matrices. Among the 20 formulations analysed, two were strictly herbal and four had a composition corresponding to declared ingredients on the packaging or the leaflet. The others were all adulterated. Eight formulations contain sibutramine alone at doses ranging from 4.4 to 30.5 mg/capsule. Five formulations contain sibutramine (from 5.0 to 19.6 mg/capsule or tablet) in combination with phenolphthalein (from 4.4 to 66.1 mg/capsule), and the last formulation was adulterated with synephrine (19.5 mg/capsule). Quantification of the actives was carried out with 1H-NMR. Several other compounds were also characterized including methylsynephrine, vitaberin, sugars, vitamins, etc. DOSY NMR is thus proposed as a useful tool for detection of unexpected adulteration.


Subject(s)
Anti-Obesity Agents/analysis , Anti-Obesity Agents/chemistry , Dietary Supplements/analysis , Drug Contamination , Drugs, Chinese Herbal/chemistry , Food Contamination , Magnetic Resonance Spectroscopy/methods , Cyclobutanes/analysis , Cyclobutanes/chemistry , Drug and Narcotic Control/methods , Limit of Detection , Pharmaceutic Aids/analysis , Pharmaceutic Aids/chemistry , Phenolphthalein/analysis , Phenolphthalein/chemistry , Synephrine , Tandem Mass Spectrometry
7.
Anal Bioanal Chem ; 398(1): 77-92, 2010 Sep.
Article in English | MEDLINE | ID: mdl-20437031

ABSTRACT

In recent years, the number of counterfeit drugs has increased dramatically, including not only "lifestyle" products but also vital medicines. Besides the threat to public health, the financial and reputational damage to pharmaceutical companies is substantial. The lack of robust information on the prevalence of fake drugs is an obstacle in the fight against drug counterfeiting. It is generally accepted that approximately 10% of drugs worldwide could be counterfeit, but it is also well known that this number covers very different situations depending on the country, the places where the drugs are purchased, and the definition of what constitutes a counterfeit drug. The chemical analysis of drugs suspected to be fake is a crucial step as counterfeiters are becoming increasingly sophisticated, rendering visual inspection insufficient to distinguish the genuine products from the counterfeit ones. This article critically reviews the recent analytical methods employed to control the quality of drug formulations, using as an example artemisinin derivatives, medicines particularly targeted by counterfeiters. Indeed, a broad panel of techniques have been reported for their analysis, ranging from simple and cheap in-field ones (colorimetry and thin-layer chromatography) to more advanced laboratory methods (mass spectrometry, nuclear magnetic resonance, and vibrational spectroscopies) through chromatographic methods, which remain the most widely used. The conclusion section of the article highlights the questions to be posed before selecting the most appropriate analytical approach.


Subject(s)
Chemistry Techniques, Analytical/methods , Fraud/prevention & control , Fraud/trends , Pharmaceutical Preparations/analysis , Humans
8.
Curr Drug Deliv ; 5(1): 7-19, 2008 Jan.
Article in English | MEDLINE | ID: mdl-18220546

ABSTRACT

Proliferative vitreoretinopathy (PVR), the most serious complication causing retinal detachment surgery to fail, is one of the leading causes of vision-loss in developed countries. The pharmaceutical treatment of this disease, located in the posterior segment of the eye, is problematic because it is difficult to achieve effective drug levels in the vitreous and the retina through conventional forms of administration (topical or systemic). Intravitreal injections can deliver drugs to the retina without the side-effects associated with systemic administration. However, because PVR is a long-term complication and the half-life of most drugs in the vitreous cavity is short, repeated injections are needed but this can cause complications. Recent advances in ocular drug delivery methods and the development of novel bioactive compounds could lead to new ways for the treatment of PVR. This review will summarize recent literature concerning intraocular drug delivery of biopharmaceutical agents for the treatment and prevention of PVR.


Subject(s)
Anti-Inflammatory Agents/administration & dosage , Antimetabolites, Antineoplastic/administration & dosage , Drug Delivery Systems/methods , Silicone Oils , Vitreoretinopathy, Proliferative/drug therapy , Animals , Anti-Inflammatory Agents/therapeutic use , Antimetabolites, Antineoplastic/therapeutic use , Colloids , Delayed-Action Preparations , Drug Implants , Humans , Injections , Liposomes , Microspheres , Ophthalmic Solutions , Rabbits , Vitreous Body
9.
J Mol Cell Cardiol ; 42(3): 526-39, 2007 Mar.
Article in English | MEDLINE | ID: mdl-17222424

ABSTRACT

Obesity is a risk factor for heart failure through a set of hemodynamic and hormonal adaptations, but its contribution at the molecular level is not clearly known. Therefore, we investigated the kinetic cardiac transcriptome and metabolome in the Spontaneous Hypertensive Heart Failure (SHHF) rat. The SHHF rat is devoid of leptin signaling when homozygous for a mutation of the leptin receptor (ObR) gene. The ObR-/- SHHF rat is obese at 4 months of age and prone to heart failure after 14 months whereas its lean counterpart ObR-/+ is prone to heart failure after 16 months. We used a set of rat pangenomic high-density macroarrays to monitor left ventricle cardiac transcriptome regulation in 4- and 10-month-old, lean and obese animals. Comparative analysis of left ventricle of 4- and 10-month-old lean rat revealed 222 differentially expressed genes while 4- and 10-month-old obese rats showed 293 differentially expressed genes. (1)H NMR analysis of the metabolome of left ventricular extracts displayed a global decrease of metabolites, except for taurine, and lipid concentration. This may be attributed to gene expression regulation and likely increased extracellular mass. The glutamine to glutamate ratio was significantly lower in the obese group. The relative unsaturation of lipids increased in the obese heart; in particular, omega-3 lipid concentration was higher in the 10-month-old obese heart. Overall, several specific kinetic molecular patterns act as a prelude to heart failure in the leptin signaling deficient SHHF obese rat.


Subject(s)
Glutamates/metabolism , Glutamine/metabolism , Heart Failure/metabolism , Intracellular Membranes/metabolism , Lipid Metabolism , Obesity/metabolism , Transcription, Genetic/genetics , Adaptation, Biological , Aging/physiology , Animals , Gene Expression Profiling , Heart Failure/genetics , Magnetic Resonance Spectroscopy , Male , Multigene Family , Obesity/genetics , Oligonucleotide Array Sequence Analysis , Phenotype , Rats
10.
Drug Metab Lett ; 1(2): 101-7, 2007 Apr.
Article in English | MEDLINE | ID: mdl-19356028

ABSTRACT

Glucuronidation of 5'-DFCR, a metabolite of capecitabine, was confirmed in experimental models from rats whereas 5'-DFCR glucuronide was detected neither in bile or liver from mice nor in liver microsomes from human. Metabolic interactions at the level of the glucuronidation pathway between CAP and other drugs are unlikely in patients.


Subject(s)
Antimetabolites, Antineoplastic/metabolism , Deoxycytidine/analogs & derivatives , Fluorouracil/analogs & derivatives , Glucuronides/metabolism , Animals , Bile/metabolism , Capecitabine , Deoxycytidine/metabolism , Fluorouracil/metabolism , Humans , Liver/metabolism , Male , Mice , Microsomes, Liver/metabolism , Rats , Rats, Wistar , Species Specificity
11.
Xenobiotica ; 33(3): 289-303, 2003 Mar.
Article in English | MEDLINE | ID: mdl-12637246

ABSTRACT

1. The metabolic pathway(s) of OGT 719, a novel nucleoside analogue in which galactose is covalently attached to the N1 of 5-fluorouracil (FU), have been investigated with (19)F-NMR spectroscopy in (1) the isolated perfused rat liver (IPRL) model, (2) normal rats, (3) rats bearing the HSN LV10 sarcoma, (4) nude mice xenografted with the human hepatoma HepG2 and (5) urine from patients. 2. The administration of OGT 719 results in the formation of small amounts of FU. IPRL experiments with OGT 719 in combination with asialofetuin, a natural asialoglycoprotein receptor (ASGP-r), suggest competitive binding of OGT 719 to the ASGP-r. 3. The data obtained in non-tumour rats also demonstrated an extremely low metabolization of OGT 719 into FU and alpha-fluoro-beta-alanine, the well-known major metabolite of FU. 4. A comparison of tumour extracts from rats bearing the HSN LV10 sarcoma treated with FU or OGT 719 showed the incorporation of FU into RNA in rats treated with FU but not in rats treated with OGT 719; nevertheless, the incorporation of FU into RNA was observed in the liver from rats treated with OGT 719. 5. In a human hepatoma xenografted to nude mice, both the OGT 719 and FU contents of the tumour were markedly higher than in the corresponding liver, suggesting a tumour-specific trapping of OGT 719 in hepatoma. 6. The metabolism of OGT 719 was also extremely low in patients. 7. In conclusion, the present study shows the value of (19)F-NMR for demonstrating for the first time that OGT 719 is a prodrug of FU although very poorly metabolized.


Subject(s)
Fluorouracil/analogs & derivatives , Fluorouracil/metabolism , Liver/metabolism , Neoplasms, Experimental/metabolism , Neoplasms/metabolism , Animals , Asialoglycoproteins/pharmacology , Biotransformation , Fetuins , Fluorine Radioisotopes , Fluorouracil/pharmacokinetics , Fluorouracil/urine , Glucuronidase/chemistry , Humans , Hydrolysis , In Vitro Techniques , Magnetic Resonance Spectroscopy , Mice , Mice, Nude , Rats , Rats, Wistar , Sarcoma, Experimental/metabolism , Transplantation, Heterologous , alpha-Fetoproteins/pharmacology
12.
Oncologist ; 7(4): 288-323, 2002.
Article in English | MEDLINE | ID: mdl-12185293

ABSTRACT

Although 5-fluorouracil (5-FU) was first introduced in 1957, it remains an essential part of the treatment of a wide range of solid tumors. 5-FU has antitumor activity against epithelial malignancies arising in the gastrointestinal tract and breast as well as the head and neck, with single-agent response rates of only 10%-30%. Although 5-FU is still the most widely prescribed agent for the treatment of colorectal cancer, less than one-third of patients achieve objective responses. Recent research has focused on the biomodulation of 5-FU to improve the cytotoxicity and therapeutic effectiveness of this drug in the treatment of advanced disease. As all the anticancer agents, 5-FU leads to several toxicities. The toxicity profile of 5-FU is schedule dependent. Myelotoxicity is the major toxic effect in patients receiving bolus doses. Hand-foot syndrome (palmar-plantar erythrodysesthesia), stomatitis, and neuro- and cardiotoxicities are associated with continuous infusions. Other adverse effects associated with both bolus-dose and continuous-infusion regimens include nausea and vomiting, diarrhea, alopecia, and dermatitis. All these reasons explain the need for more effective and less toxic fluoropyrimidines. In the first part of this review, we briefly present the metabolic pathways of 5-FU responsible for the efficacy and toxicity of this drug. This knowledge is also necessary to understand the target(s) of biomodulation. The second part is devoted to a review of the literature on three recent prodrugs of 5-FU, i.e., capecitabine, UFT (ftorafur [FTO] plus uracil), and S-1 (FTO plus 5-chloro-2,4-dihydroxypyridine plus potassium oxonate). The pharmacological principles that have influenced the development of these new drugs and our current knowledge of the clinical pharmacology of these new agents, focusing on antitumor activity and toxicity, are presented. The literature was analyzed until March 2002. This review is intended to be as exhaustive as possible since it was conceived as a work tool for readers wanting to go further.


Subject(s)
Antineoplastic Agents/therapeutic use , Deoxycytidine/analogs & derivatives , Deoxycytidine/therapeutic use , Fluorouracil/therapeutic use , Prodrugs/therapeutic use , Pyridines/therapeutic use , Tegafur/therapeutic use , Uracil/therapeutic use , Administration, Oral , Antineoplastic Agents/metabolism , Antineoplastic Agents/toxicity , Capecitabine , Deoxycytidine/metabolism , Deoxycytidine/toxicity , Fluorouracil/metabolism , Fluorouracil/toxicity , Humans , Neoplasms/drug therapy , Prodrugs/metabolism , Prodrugs/toxicity , Pyridines/metabolism , Pyridines/toxicity , Tegafur/metabolism , Tegafur/toxicity , Treatment Outcome , Uracil/metabolism , Uracil/toxicity
13.
Curr Med Chem Anticancer Agents ; 2(2): 267-310, 2002 Mar.
Article in English | MEDLINE | ID: mdl-12678747

ABSTRACT

Although 5-fluorouracil (FU) was first introduced in 1957, it remains an essential part of the treatment of a wide range of solid tumors. FU has antitumor activity against epithelial malignancies arising in the gastrointestinal tract, breast as well as the head and neck, with single-agent response rates of only 10-30%. Although FU is still the most widely prescribed agent for the treatment of colorectal cancer, less than a third of patients achieve objective responses. Recent research has focused on the biomodulation of FU to improve the cytotoxicity and therapeutic effectiveness of this drug in the treatment of advanced disease. As all the anticancer agents, FU leads to several toxicities. The toxicity profile of FU is schedule dependent. Myelotoxicity is the major toxic effect in patients receiving bolus doses. Hand-foot syndrome (palmar-plantar erythrodysesthesia), stomatitis, neuro- and cardiotoxicity are associated with continuous infusions. Other adverse effects associated with both bolus-dose and continuous infusion regimens include nausea and vomiting, diarrhea, alopecia and dermatitis. All these reasons explain the need of more effective and less toxic fluoropyrimidines. In the first part of the review, we briefly present the metabolic pathways of FU responsible for the efficacy and toxicity of the drug. This knowledge is also necessary to understand the target(s) of the biomodulation. The second part is devoted to a review of the literature on the various prodrugs of FU, including 5'-deoxy-5-fluorouridine, capecitabine, BOF-A2, ftorafur, UFT, and S-1. The promising approach of gene directed enzyme-prodrug therapy is also presented. A brief survey of antibody directed enzyme-prodrug therapy and some new FU prodrugs concludes the paper. The pharmacological principles that have influenced the development of these new drugs and our current knowledge of the clinical pharmacology of these new agents, focusing on antitumor activity and toxicity, are presented.


Subject(s)
Antimetabolites, Antineoplastic/chemical synthesis , Fluorouracil/chemical synthesis , Prodrugs/chemical synthesis , Animals , Antimetabolites, Antineoplastic/pharmacokinetics , Antimetabolites, Antineoplastic/pharmacology , Antimetabolites, Antineoplastic/therapeutic use , Biotransformation , Fluorouracil/pharmacokinetics , Fluorouracil/pharmacology , Fluorouracil/therapeutic use , Humans , Neoplasms/drug therapy , Neoplasms/pathology , Prodrugs/pharmacology , Prodrugs/therapeutic use
14.
Rev Neurol (Paris) ; 157(8-9 Pt 1): 858-62, 2001 Sep.
Article in French | MEDLINE | ID: mdl-11677409

ABSTRACT

Magnetic resonance spectroscopy (MRS) is a tool for a non-invasive monitoring of brain tumor metabolism. In vivo proton MRS became possible with the development of whole-body high-field magnets. First, it allows to distinguish brain tumors from abscesses. Second, along with other imaging techniques, it permits the differentiation of primary brain tumors, mainly gliomas, from tumors of various origins such as meningiomas or metastasis. However, its ability to give a grading of gliomas stays controversial. Choline, a marker of cell membrane proliferation, could give information on the degree of malignancy but reports on its role are somewhat contradictory. Brain tumor biopsy and histology stay mandatory in the management of brain tumors. In vitro MRS spectra obtained from tumor extracts show that the signal at 3.2ppm, the so-called "cholin peak", corresponds to several compounds among them, glycerophosphocholine, phosphocholine, and choline. Their repartition differs with the grade of the tumor. In vivo proton MRS is the only metabolic technique of non-invasive monitoring of treated brain tumors. It can separate recurrence from radionecrosis. Improved methodology and availability of MR imagers will strengthen its importance in the future.


Subject(s)
Aspartic Acid/analogs & derivatives , Brain Neoplasms/diagnosis , Energy Metabolism/physiology , Glioma/diagnosis , Magnetic Resonance Spectroscopy , Aspartic Acid/metabolism , Biopsy , Brain/pathology , Brain/physiopathology , Brain Neoplasms/pathology , Brain Neoplasms/physiopathology , Brain Neoplasms/secondary , Choline/metabolism , Diagnosis, Differential , Glioma/physiopathology , Humans , Predictive Value of Tests , Prognosis
15.
J Pharm Biomed Anal ; 25(3-4): 669-78, 2001 Jun.
Article in English | MEDLINE | ID: mdl-11377048

ABSTRACT

NMR (31P, 1H and 13C) spectroscopy was used to study the products of the degradation of isophosphoramide mustard (IPM) in buffered solutions at pH ranging from 1 to 13. At pH < or = 1, the only degradation compounds detected were phosphate ion (Pi) and chloroethylammonium chloride (CEA-HCl), resulting from the breakdown of the two P-N bonds (pathway Ia). At pH 9.3 and 13, only the products of 1,3-cyclization of the N-chloroethyl group (monoaziridinylIPM (monoAzIPM) and a very low level of bisaziridinylIPM (bisAzIPM)) were found after approximately 15 h of reaction (pathway II). At intermediate pH, the two pathways coexist. At pH 3.5 and 5.0, the P-N bond hydrolysis is the major pathway, but two final phosphorylated products were detected, Pi which represented 67% (pH 3.5) and 17% (pH 5.0) of all the IPM phosphorylated degradation products after approximately 15 h of reaction, and phosphorylethanolamine (PEA) which represented 16% (pH 3.5) and 46% (pH 5.0) of the same sum. PEA formation can be explained by the 1,5-cyclization of a transient compound giving a 1,3,2-oxazaphospholidine intermediate whose P-N bond is exclusively cleaved in acidic medium. The presence of monohydroxyIPM (monoOHIPM) (whose percentage increases with pH from 5% (pH 3.5) to approximately 28% (pH 5.0) of all the IPM phosphorylated degradation compounds), probably coming from the alkylation by water of an aziridine/aziridinium intermediate, demonstrates the occurrence of pathway II. At pH 7.0 and 7.4, the pathway II is initiated first, leading to 1,3-cyclization(s), followed by water alkylation of the aziridines formed. The sequences are IPM 1-->monoAzIPM 5-->bisAzIPM 9; IPM 1-->monoAzIPM 5-->monoOHIPM 6-->monoAzIPM with a N-hydroxyethylchain (presumed structure) 7-->dihydroxyIPM 8. Nevertheless, PEA and Pi are the final products observed, which implies the P-N bond hydrolysis of products 5-9 as demonstrated by the presence in the medium of CEA, aziridine and ethanolamine.


Subject(s)
Antineoplastic Agents, Alkylating/chemistry , Phosphoramide Mustards/chemistry , Drug Stability , Hydrogen-Ion Concentration , Magnetic Resonance Spectroscopy , Solutions
16.
Cancer Chemother Pharmacol ; 46(6): 501-6, 2000.
Article in English | MEDLINE | ID: mdl-11138464

ABSTRACT

PURPOSE: The stability of 5-fluorouracil (FU) Roche solutions in a portable infusion pump under prolonged "in-use" conditions (32 degrees C, in the dark) was studied, especially with respect to the formation of the cardiotoxic compounds fluoroacetaldehyde (Facet) and fluoromalonic acid semialdehyde (FMASAld). METHODS: The solutions, prepared according to three protocols frequently used at the Anticancer Centre in Toulouse, were analysed by 19F NMR immediately after preparation (T0) and after 2, 3 or 10 days (TF) in the pump. RESULTS: The commercial solution already contained 64 fluorinated "impurities", among them fluoride ion (F-), FMASAld and Facet. The concentration of FU did not change significantly between T0 and TF, whatever the protocol. The levels of F- had not increased significantly after 2 or 3 days, but had increased by about 50% after 10 days. The increases in FMASAld levels were low (12-28%) albeit significant in the three protocols. The levels of Facet had increased by a factor of about 2 after 2 or 3 days, and by a factor of > 3 after 10 days. The levels of the other fluorinated compounds were constant during the first 2 or 3 days, but had increased by about 30% after 10 days. FU Dakota lyophilizates, analysed immediately after reconstitution, contained neither FMASAld nor Facet. After 2 days at 25 degrees C, low levels of FMASAld were present but Facet could still not be detected. CONCLUSION: This study showed that special attention must be paid to the risk of increasing concentrations of highly toxic FMASAld and Facet when FU is administered via a pump for long periods of time. It would be preferable not to exceed 3 days of treatment when patients receive FU from a portable infusion pump. This underlines the interest in using a lyophilized formulation of FU in clinical practice.


Subject(s)
Antimetabolites, Antineoplastic/chemistry , Fluorouracil/chemistry , Infusion Pumps , Antimetabolites, Antineoplastic/administration & dosage , Drug Stability , Fluorine Radioisotopes , Fluorouracil/administration & dosage , Hydrogen-Ion Concentration , Magnetic Resonance Spectroscopy , Time Factors
17.
Curr Drug Metab ; 1(3): 271-303, 2000 Nov.
Article in English | MEDLINE | ID: mdl-11465049

ABSTRACT

Fluorine-19 nuclear magnetic resonance (19F NMR) spectroscopy provides a highly specific tool for identifying fluorine-containing drugs and their metabolites in biological media. This article focuses on the application of 19F NMR to the metabolic studies of fluoropyrimidine drugs in clinical use. The value and difficulties encountered in investigations on drug metabolism are first discussed. The metabolism and disposition studies of the anticancer drug 5-fluorouracil, the mainstay of antimetabolite treatment for solid tumors, and its prodrugs, doxifluridine and capecitabine, are then extensively reviewed. The studies dealing with the antimycotic agent, 5-fluorocytosine, as well as the novel anticancer drug, gemcitabine, are also considered. From in vitro (biofluids or tissue extracts) 19F NMR analysis, seven new metabolites of 5-fluorouracil, doxifluridine, capecitabine and 5-fluorocytosine were identified. Except two, they were only detected using this technique. This emphasizes the high analytical potential of in vitro 19F NMR. In vivo 19F NMR is non-invasive and thus allows the quantitative monitoring of the metabolism of 5-fluorouracil in the target tissue, e.g. the tumor, as well as its biodistribution. Another promising application is its ability to estimate the level of yeast cytosine deaminase gene expression in human tumors from the quantitative monitoring of 5-fluorouracil formation from the non-cytotoxic drug 5-fluorocytosine. Notwithstanding these successes, the limited sensitivity and spectral resolution of 19F NMR precludes its extensive applicability to all the fluorinated drugs.


Subject(s)
Fluorine/chemistry , Magnetic Resonance Spectroscopy/methods , Pyrimidines/chemistry , Animals , Antimetabolites, Antineoplastic/chemistry , Antimetabolites, Antineoplastic/metabolism , Fluorouracil/analogs & derivatives , Fluorouracil/chemistry , Fluorouracil/metabolism , Humans , Pyrimidines/metabolism
18.
Curr Pharm Des ; 5(8): 561-86, 1999 Aug.
Article in English | MEDLINE | ID: mdl-10469892

ABSTRACT

Cyclophosphamide (CP) has been in clinical use for the treatment of malignant disease for over 40 years. CP is inactive until it undergoes complex metabolic pathways leading to the ultimate alkylating agent, phosphoramide mustard, but also to inactive and toxic metabolites. Sensitive and specific methods are now available for the measurement of CP and its enantiomers, its metabolites and their stereoisomers, in biological matrices. An overview is given of the methods of analysis of CP and its metabolites described in literature since 1993 as well as the current knowledge about its metabolism. Five classes of methods are described: (1) thin-layer chromatography-photographic densitometry, (2) high performance liquid chromatography, (3) gas chromatography and gas chromatography coupled to mass spectrometry, (4) phosphorus-31 nuclear magnetic resonance and (5) enantiomeric separation. In each case, sample clean up and preparation are described. Precision and limits of quantification of the assays are indicated. A table summarizes all the analytical methods for assaying each metabolite.


Subject(s)
Antineoplastic Agents, Alkylating/metabolism , Cyclophosphamide/metabolism , Antineoplastic Agents, Alkylating/analysis , Antineoplastic Agents, Alkylating/history , Cyclophosphamide/analysis , Cyclophosphamide/history , History, 20th Century , Humans
19.
J Med Chem ; 42(14): 2542-60, 1999 Jul 15.
Article in English | MEDLINE | ID: mdl-10411475

ABSTRACT

31P NMR spectroscopy was used to study the products of the decomposition of the antitumor drug ifosfamide (IF, 1d) and its N-dechloroethylated metabolites, namely, 2,3-didechloroethylIF (1a) and 2- (1b) and 3-dechloroethylIF (1c), in buffered solutions at acidic pH. The first stage of acid hydrolysis of these four oxazaphosphorines is a P-N bond cleavage of the six-membered ring leading to the phosphoramidic acid monoesters (2a-d) of type R'HN(CH(2))(3)OP(O)(OH)NHR, with R and/or R' = H or (CH(2))(2)Cl. The electron-withdrawing chloroethyl group at the endocyclic and/or exocyclic nitrogens counteracts the endocyclic P-N bond hydrolysis. This effect is even more marked when the N-chloroethyl group is in the exocyclic position since the order of stability is 1d > 1c > 1b > 1a. In the second stage of hydrolysis, the remaining P-N bond is cleaved together with an intramolecular attack at the phosphorus atom by the non-P-linked nitrogen of the compounds 2a-d. This leads to the formation of a 2-hydroxyoxazaphosphorine ring with R = H (3a coming from compounds 2a,c) or (CH(2))(2)Cl (3b coming from compounds 2b,d) and to the release of ammonia or chloroethylamine. The third step is the P-N ring opening of the oxazaphosphorines 3a,b leading to the phosphoric acid monoesters, H(2)N(CH(2))(3)OP(O)(OH)(2) (4a) and Cl(CH(2))(2)HN(CH(2))(3)OP(O)(OH)(2) (4b-1), respectively. For the latter compound, the chloroethyl group is partially (at pH 5.5) or totally (at pH 7.0) cyclized into aziridine (4b-2), which is then progressively hydrolyzed into an N-hydroxyethyl group (4b-3). Compounds 3a,b are transient intermediates, which in strongly acidic medium are not observed with (31)P NMR. In this case, cleavage of the P-N bond of the type 2 phosphoramidic acid monoesters leads directly to the type 4 phosphoric acid monoesters. The phosphate anion, derived from P-O bond cleavage of these latter compounds, is only observed at low levels after a long period of hydrolysis. Compounds 1a-c and some of their hydrolytic degradation products (4b-1, 4b-2, diphosphoric diester [Cl(CH(2))(2)NH(CH(2))(3)OP(O)(OH)](2)O (5), and chloroethylamine) did not exhibit, as expected, any antitumor efficacy in vivo against P388 leukemia. (31)P NMR determination of the N-dechloroethylated metabolites of IF or its structural isomer, cyclophosphamide (CP), and their degradation compounds could provide an indirect and accurate estimation of chloroacetaldehyde amounts formed from CP or IF.


Subject(s)
Antineoplastic Agents/chemistry , Ifosfamide/analogs & derivatives , Ifosfamide/chemistry , Phosphoramide Mustards/chemistry , Amides/chemistry , Animals , Antineoplastic Agents/pharmacology , Antineoplastic Agents/toxicity , Antineoplastic Agents/urine , Drug Stability , Female , Hydrogen-Ion Concentration , Hydrolysis , Ifosfamide/pharmacology , Ifosfamide/toxicity , Ifosfamide/urine , Kinetics , Lethal Dose 50 , Leukemia P388/drug therapy , Magnetic Resonance Spectroscopy , Male , Mice , Neoplasm Transplantation , Phosphoramide Mustards/pharmacology , Phosphoramide Mustards/toxicity , Phosphoric Acids/chemistry , Phosphorus Isotopes , Rats , Tumor Cells, Cultured , Water
20.
Invest Radiol ; 34(3): 230-5, 1999 Mar.
Article in English | MEDLINE | ID: mdl-10084669

ABSTRACT

RATIONALE AND OBJECTIVES: The authors sought to compare 1H magnetic resonance spectroscopy (MRS) spectra from extracts of low-grade and high-grade gliomas, especially with respect to the signals of choline-containing compounds. METHODS: Perchloric acid extracts of six high-grade and six low-grade gliomas were analyzed by 1H MRS at 9.4 Tesla. RESULTS: The signals of glycerophosphocholine (GPC) at 3.23 ppm, phosphocholine (PC) at 3.22 ppm, and choline (Cho) at 3.21 ppm were identified in both types of tumors. The absolute concentrations of all Cho-containing compounds (GPC + PC + Cho) in high-grade and low-grade gliomas were significantly different. The relative contributions of each of the Cho-containing compounds to the total choline signal were also statistically different. For high-grade gliomas, the choline signal is composed of GPC, PC, and Cho in a well-balanced contribution, whereas in low-grade gliomas, the signal is largely due to GPC with a small involvement of PC and Cho. CONCLUSIONS: The differences in the concentration and the repartition of Cho-containing compounds seem to be a marker of high-grade gliomas. They could also help to discriminate between high- and low-grade gliomas in some difficult cases, especially if there is histologic uncertainty between anaplastic astrocytomas and low-grade oligodendrogliomas.


Subject(s)
Brain Neoplasms/metabolism , Choline/metabolism , Glioma/metabolism , Magnetic Resonance Imaging , Adult , Aged , Brain Neoplasms/pathology , Female , Glioma/pathology , Humans , In Vitro Techniques , Male , Middle Aged , Phosphorylcholine/metabolism
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