RESUMEN
BACKGROUND: Dasatinib, as an oral multi-targeted inhibitor of BCR-ABL and SRC family kinases, has been widely used for the treatment of Philadelphia Chromosome Positive Leukemias in imatinib-acquired resistance and intolerance. The study aimed to develop and validate a simple and robust assay with a small volume of plasma based on liquid chromatography coupled with tandem mass spectrometry to determine the concentration of dasatinib and to investigate the impact of the cytochrome 3A4 inhibitors, including ketoconazole, voriconazole, itraconazole and posaconazole, on the pharmacokinetics of dasatinib in rats. METHODS: Thirty rats were divided randomly into five groups, control group (0.5% carboxymethylcellulose sodium), ketoconazole (30 mg/kg) group, voriconazole group (30 mg/kg), itraconazole group (30 mg/kg) and posaconazole group (30 mg/kg). After 150 µL blood samples were collected at 0, 0.5, 1, 2, 4, 6, 8, 10, 12, 24, and 48 h and precipitated with acetonitrile, the plasma concentration of dasatinib was determined through Fluoro- Phenyl column (150 mm×2.1 mm, 3 µm) in a positive ionization mode. RESULTS: The results suggested that ketoconazole, voriconazole, and posaconazole could increase the AUC0-t of dasatinib to varying degrees while significantly reducing its clearance. However, there was no significant impact on the pharmacokinetics of dasatinib, co-administered with itraconazole except for the CL and MRT0-t of dasatinib. Additionally, voriconazole could significantly increase Cmax of dasatinib by approximately 4.12 fold. CONCLUSION: These data indicated that ketoconazole, posaconazole and voriconazole should be cautiously co-administered with dasatinib or close therapeutic drug monitoring of dasatinib concentration, which might cause the drug-drug interaction.
Asunto(s)
Antifúngicos/farmacocinética , Inhibidores del Citocromo P-450 CYP3A/farmacocinética , Dasatinib/farmacocinética , Monitoreo de Drogas/métodos , Leucemia Mielógena Crónica BCR-ABL Positiva/tratamiento farmacológico , Inhibidores de Proteínas Quinasas/farmacocinética , Administración Oral , Animales , Antifúngicos/administración & dosificación , Antifúngicos/aislamiento & purificación , Área Bajo la Curva , Cromatografía Líquida de Alta Presión/métodos , Citocromo P-450 CYP3A/metabolismo , Inhibidores del Citocromo P-450 CYP3A/administración & dosificación , Inhibidores del Citocromo P-450 CYP3A/aislamiento & purificación , Dasatinib/administración & dosificación , Dasatinib/aislamiento & purificación , Interacciones Farmacológicas , Humanos , Itraconazol/administración & dosificación , Itraconazol/aislamiento & purificación , Itraconazol/farmacocinética , Cetoconazol/administración & dosificación , Cetoconazol/aislamiento & purificación , Cetoconazol/farmacocinética , Masculino , Modelos Animales , Inhibidores de Proteínas Quinasas/administración & dosificación , Ratas , Espectrometría de Masas en Tándem/métodos , Triazoles/administración & dosificación , Triazoles/aislamiento & purificación , Triazoles/farmacocinética , Voriconazol/administración & dosificación , Voriconazol/aislamiento & purificación , Voriconazol/farmacocinéticaRESUMEN
The addition of matrix formers within a formulation provides a means for enhancing the redispersibility of nanoparticles (NPs) enabling them to retain their advantageous properties imparted onto them by their sub-micron size. In this work, NPs were isolated in the solid state via spray drying with a range of sugars. The processed powders were characterized, establishing that itraconazole (ITR) nanostructured microparticles (NMPs) spray dried in the presence of mannitol and trehalose had favorable redispersibility confirmed by dynamic light scattering and nanoparticle tracking analysis. Solid-state analysis confirmed the crystalline nature of NMPs based on mannitol and the amorphous character of trehalose-based NMPs. The NMPs powders were compacted at a range of pressures, producing tablets with high tensile strength without compromising their disintegration time. A greater amount of ITR was solubilized from trehalose NMPs compared to the mannitol-based compacts in 0.1 M HCl, showing a promise for enhanced in vivo activity. Overall, as trehalose exhibited superior carrier properties for ITR NMPs, this type of excipient included in the formulation warrants careful consideration. The structured approach to matrix former selection and tabletting studies can reduce the amount of material and time required for testing in the initial stages of product development.
Asunto(s)
Itraconazol/aislamiento & purificación , Nanoestructuras/química , Desecación , Excipientes/química , Itraconazol/química , Manitol/química , Tamaño de la Partícula , Polvos , Comprimidos , Resistencia a la Tracción , Trehalosa/químicaRESUMEN
Nowadays in animal studies, it is important to comply with the so-called Three Rs rule by replacing or reducing the number of tested animals. Volumetric absorptive microsampling (VAMS) can be used to collect small quantities (10 or 20µL) of whole blood, thereby limiting the amount of animals needed. In this study, a quantitative method was developed and subsequently validated for the poorly soluble drug itraconazole (ITZ) using VAMS and ultra-high performance liquid chromatography (UHPLC) coupled to tandem mass spectrometry (MS). A proof of concept study showed that the optimized method is applicable to test the bioavailability of drug formulations containing ITZ. Using VAMS, smaller blood volumes can be taken per sampling point (10-20µL instead of the conventional 0.2-0.5mL) avoiding the sacrifice of animals. Moreover, the same rats can be used to compare different drug formulations which strengthens the validity of the results. In long-term bioavailability studies, it is necessary to guarantee the stability of the tested drugs supported on VAMS devices. In this study, we show that ITZ was only stable for 24h after collection with VAMS, but for at least two weeks by the storage of extracted samples at -80°C.
Asunto(s)
Itraconazol/sangre , Espectrometría de Masas en Tándem , Animales , Disponibilidad Biológica , Cromatografía Líquida de Alta Presión , Composición de Medicamentos , Semivida , Itraconazol/aislamiento & purificación , Itraconazol/farmacocinética , Masculino , Ratas , Ratas Wistar , Solventes/químicaRESUMEN
cis-Itraconazole is a chiral antifungal drug administered as a racemate. The knowledge of properties of individual cis-itraconazole stereoisomers is vital information for medicine and biosciences as different stereoisomers of cis-itraconazole may possess different affinity to certain biological pathways in the human body. For this purpose, either chiral synthesis of enantiomers or chiral separation of racemate can be used. This paper presents a two-step high-performance liquid chromatography approach for the semipreparative isolation of four stereoisomers (two enantiomeric pairs) of itraconazole using polysaccharide stationary phases and volatile organic mobile phases without additives in isocratic mode. The approach used involves the separation of the racemate into three fractions (i.e. two pure stereoisomers and one mixed fraction containing the remaining two stereoisomers) in the first run and consequent separation of the collected mixed fraction in the second one. For this purpose, combination of cellulose tris-(4-methylbenzoate) and cellulose tris-(3,5-dimehylphenylcarbamate) columns with complementary selectivity for cis-itraconazole provided full separation of all four stereoisomers (with purity of each isomer > 97%). The stereoisomers were collected, their optical rotation determined and their identity confirmed based on the results of a previously published study. Pure separated stereoisomers are subjected to further biological studies.
Asunto(s)
Antifúngicos/aislamiento & purificación , Celulosa/química , Itraconazol/aislamiento & purificación , Antifúngicos/química , Cromatografía Líquida de Alta Presión , Itraconazol/química , Conformación Molecular , EstereoisomerismoRESUMEN
A chromatographic method (high-performance liquid chromatography/photodiode array detection) for the simultaneous determination of itraconazole and its mayor metabolite, hydroxy itraconazole, was developed. The validation data demonstrated acceptable values for linearity (dynamic range between 0.25-16 µg/mL for hydroxy itraconazole, 0.125-4 µg/mL for itraconazole), precision (Coefficient of Variation, 2.23-4.28% and 1.61-9.52% respectively), accuracy (Relative Error, -2.89 to -17.26% and -3.11 to 14.44%), selectivity, sensitivity (Lower limit of Quantification, 0.125 and 0.25 µg/mL), and recovery (97.30-105.82%) for both compounds. Its clinical suitability was also investigated in 104 trough human serum samples. The assay resulted to be rapid, sensitive, and simple for simultaneously quantification of these azole compounds in serum samples and should make a positive contribution to optimize therapies on an individual basis.
Asunto(s)
Antifúngicos/sangre , Itraconazol/análogos & derivados , Adulto , Antifúngicos/aislamiento & purificación , Análisis Químico de la Sangre/normas , Niño , Cromatografía Líquida de Alta Presión/normas , Estabilidad de Medicamentos , Humanos , Itraconazol/sangre , Itraconazol/aislamiento & purificación , Límite de Detección , Control de Calidad , Estándares de ReferenciaRESUMEN
Itraconazole is an antifungal drug widely used in a variety of fungal infections, which have become a significant public-health problem in recent decades. Itraconazole is a chiral drug consisting of two diastereoisomeric racemates, i.e., four stereoisomers. Data in the literature suggests that stereochemistry may play a significant role in the action and disposition of the drug and therefore stereoselective analytical methods for the determination of the drug in biological fluids are needed for the elucidation of that role. We report a stereoselective HPLC method that incorporates solvent extraction, the use of an internal standard, two chiral stationary phases in series, and fluorescence detection. The procedure is enantioselective and partially diastereoselective and provides the concentrations in blood plasma of the two epimer mixtures 2R,4S,2'R/2R,4S2'S and 2S,4R,2'R/2S,4R,2'S, respectively, each of which is a combination of the two epimers that differ in the configuration at the sec-butyl group. The analytical method has suitable sensitivity, recovery, precision, and accuracy. Analysis of the plasma of a human subject six hours after the oral administration of a single 200-mg dose of itraconazole showed a 3.4-fold difference between the concentrations of the epimer mixtures. The method has certain advantages over the published alternative procedure that uses LC-MS.
Asunto(s)
Análisis Químico de la Sangre/métodos , Itraconazol/sangre , Itraconazol/química , Análisis Químico de la Sangre/normas , Cromatografía Líquida de Alta Presión , Humanos , Itraconazol/aislamiento & purificación , Límite de Detección , Modelos Lineales , Estándares de Referencia , Espectrometría de Fluorescencia , EstereoisomerismoRESUMEN
BACKGROUND: A steady-state trough plasma itraconazole concentration greater than 500 ng/mL is a therapeutic target for itraconazole. A simple, rapid and sensitive high-performance liquid chromatography-based method was developed for quantitation of itraconazole and hydroxyitraconazole in human plasma. METHODS: Itraconazole and hydroxyitraconazole were separated using a mobile phase of 0.5% KH2PO4 (pH 6.0)-acetonitrile (30:70, v/v) on a CAPCELLPAK C18 MGII column at a flow rate of 0.5 mL/min and ultraviolet absorbance at 260 nm. RESULTS: The analysis required 200 microL of plasma and involved a rapid, simple solid-phase extraction with an Oasis HLB cartridge, which resulted in recoveries of 87-92% for itraconazole and 91-94% for hydroxyitraconazole. The lower limit of quantification for itraconazole and hydroxyitraconazole was 5 ng/mL each. Intra- and interday coefficients of variation for itraconazole and hydroxyitraconazole were less than 11.3% and 12.2%, respectively, and accuracies were within 11.7% and 4.5% over the linear range, respectively. Although the steady-state plasma concentrations of itraconazole and hydroxyitraconazole ranged from 506 to 2482 ng/mL and from 766 to 2444 ng/mL, respectively, after a two-day loading dose of 400 mg/day intravenous itraconazole followed by the administration of 200 mg/day itraconazole oral solution, calibration curves of itraconazole and hydroxyitraconazole showed positive linearity in a concentration range of 5-2500 and 50-2500 ng/mL, respectively. CONCLUSIONS: Our results indicate that this method is applicable for the monitoring of plasma levels of itraconazole and hydroxyitraconazole in a clinical setting. Furthermore, the regimen presented here might also be effective in preventing infection, but further studies with large sample sizes are necessary to investigate this avenue.
Asunto(s)
Análisis Químico de la Sangre/métodos , Cromatografía Líquida de Alta Presión/métodos , Itraconazol/análogos & derivados , Itraconazol/sangre , Itraconazol/farmacocinética , Rayos Ultravioleta , Calibración , Estabilidad de Medicamentos , Humanos , Itraconazol/aislamiento & purificación , Extracción en Fase SólidaRESUMEN
The four stereoisomers of itraconazole were resolved for the first time by EKC using a CD as chiral selector. A study on the enantiomeric separation ability of different neutral CDs was carried out. Heptakis-2,3,6-tri-O-methyl-beta-CD was shown to provide the highest values for the enantiomeric resolution. The influence of some experimental conditions, such as pH, chiral selector concentration, and temperature, on the enantiomeric separation was also studied. The use of a 100 mM phosphate buffer (pH 2.5), 30 mM in heptakis-2,3,6-tri-O-methyl-beta-CD together with an applied voltage of 30 kV and a temperature of 20 degrees C enabled the separation of the enantiomers of itraconazole with high resolutions (Rs > 3.0). Finally, the method was validated and successfully applied to the quantitation of itraconazole in three pharmaceutical formulations.
Asunto(s)
Antifúngicos/análisis , Electroforesis Capilar/métodos , Itraconazol/análisis , beta-Ciclodextrinas/química , Antifúngicos/química , Antifúngicos/aislamiento & purificación , Cromatografía/métodos , Formas de Dosificación , Itraconazol/química , Itraconazol/aislamiento & purificación , EstereoisomerismoRESUMEN
BACKGROUND: A practical, simple, high-performance liquid chromatographic (HPLC) method is needed for the determination of itraconazole in clinical plasma samples. METHODS: Itraconazole and bifonazole (internal standard) were extracted from plasma using a C8-bonded solid-phase cartridge, separated by C8 reversed-phase HPLC, and quantified by ultraviolet absorption at 263 nm. RESULTS: This new method enabled the determination of itraconazole in the concentration range of 10.0-500.0 microg/L. The detection limit of itraconazole was 5.0 microg/L. The mean recovery of itraconazole added to plasma was more than 89.1%, with a coefficient of variation of less than 6.9%. We applied this method for the determination of plasma itraconazole in volunteers treated daily with a 200 mg oral capsule of itraconazole for four days. We monitored the plasma level of itraconazole for the following 24 h and obtained the mean area under the plasma concentration-time curve from 0 to 24 h (AUC(0-24) ) value of 4358.9 +/- 1933.4 microg h/L. CONCLUSION: Our new method will be clinically useful for accurately monitoring the plasma concentration of itraconazole in patients under treatment.
Asunto(s)
Itraconazol/sangre , Adulto , Cromatografía Líquida de Alta Presión , Monitoreo de Drogas , Humanos , Itraconazol/aislamiento & purificación , Itraconazol/farmacocinética , Masculino , Persona de Mediana Edad , Reproducibilidad de los ResultadosRESUMEN
Itraconazole (ITC) is a hydrophobic antimycotic drug with three chiral centers that is used clinically as a stereoisomeric mixture. A chiral capillary electrophoretic method for the separation of ITC stereoisomers and those of its main metabolite hydroxyitraconazole (HITC) was developed to determine the stereoselective nature of the ITC to HITC biotransformation. The method is based on the formation of inclusion complexes of the target analytes with the negatively charged sulfated beta-cyclodextrin in the presence of moderate concentrations of methanol in a low-pH phosphate buffer. The addition of polyethylene glycol 4000 was found to be critical in obtaining baseline resolution of eight peaks, two from ITC, four from HITC, and two from R051012 (internal standard), in under 20 min. Application of the developed procedure to serum samples from patients being treated with ITC showed clearly the presence of a stereoselective component in the metabolism of this antimycotic drug. This could be shown from in vitro incubations with single enzyme Supersomes to be in part due to the stereoselective formation of HITC by the human CYP3A4 enzyme. For one patient, monitoring of the ITC and HITC concentrations and peak ratios over a 103 day period of treatment with ITC showed a strong dependency of the chiral ITC ratio to the concentration of ITC, while the dominant enantiomeric ratio of HITC was largely independent of the total HITC concentration.
Asunto(s)
Electroforesis Capilar/métodos , Itraconazol/análogos & derivados , Itraconazol/sangre , Itraconazol/aislamiento & purificación , beta-Ciclodextrinas , Ciclodextrinas , Citocromo P-450 CYP3A , Sistema Enzimático del Citocromo P-450/metabolismo , Monitoreo de Drogas , Humanos , Metanol , Polietilenglicoles , EstereoisomerismoRESUMEN
A simple and robust solid-phase extraction (SPE) procedure for the cleanup and sample preconcentration of antifungals (ketoconazole, clotrimazole, itraconazole, fluconazole, and voriconazole) and their metabolites after incubation with human liver microsomes, as well as a simplified capillary zone electrophoresis (CZE) method for their rapid analysis, have been developed to determine the stability of these compounds in in vitro samples. Three different sample pretreatment procedures using SPE with reversed-phase sorbents (100 mg C8, 100 mg C18, and 30 mg Oasis-HLB) were studied. The highest and most reproducible recoveries were obtained using a 30 mg Oasis-HLB sorbent and methanol containing 2% acetic acid as eluent. Enrichment by a factor of about four times was achieved by reconstituting the final SPE eluates to a small volume. For the CZE separation, good separations without interfering peaks due to the in vitro matrix were obtained with a simple running electrolyte using a fused-silica capillary. The best separation for all components originated by each tested drug after incubation with human liver microsomes (unmetabolized parent drug and its metabolites) was obtained using a 0.05 M phosphate running buffer (pH 2.2) without additives. The effect of the injection volume was also investigated in order to obtain the best sensitivity. Performance levels in terms of precision, linearity, limits of detection, and robustness were determined.