Hydroxy Itraconazole Research Articles

Comprehensive evaluation of solvent in dispersive liquid-liquid microextraction for determination of itraconazole and hydroxy itraconazole by high performance liquid chromatography with fluorescence detection

This paper aims to propose a multicriteria decision analysis (MCDA) method to evaluate and select solvent in a dispersive liquid-liquid microextraction (DLLME) approach. The DLLME is applied to the determination of itraconazole and hydroxy itraconazole in plasma by high performance liquid chromatography with fluorescence detection (HPLC-FLD). To achieve this goal, extraction efficiency, chromatographic resolution, and greenness of solvent were identified as the three indicators in MCDA. Then, Technique for Order of Preference by Similarity to Ideal Solution (TOPSIS) was employed to evaluate and select the solvent. Weight assignment was set up by integrating the subjective scoring from experienced analysts and the objective approach using Shannon’s entropy weight method. Under chosen parameters (extraction solvent: 1-decanol 40 μL, dispersing solvent: methanol 400 μL), the method was validated satisfactorily and applied successfully to the analysis of itraconazole and hydroxy itraconazole in real human plasma samples. The results show that the comprehensive weighted TOPSIS analysis is a promising tool to choose experimental conditions toward an integrative goal. It can provide analysts with a decision making reference to maintain a balance between the analytical performance and eco-friendliness.

Bioequivalence study of two formulations of itraconazole 100 mg capsules in healthy volunteers under fed conditions: a randomized, three-period, reference-replicated, crossover study

ABSTRACTBackground: The aim of the study was to evaluate the bioequivalence of two itraconazole 100 mg capsule formulations.Research design and methods: The single-center, open-label, randomized, three-period, three-sequence, reference-replicated, cross-over study included 38 healthy subjects under fed conditions. In each study period (separated by a 14-day washout), a single oral dose of the test (T) or reference (R) product was administered. Blood samples were collected at pre-dose and up to 72.0 h after administration. The calculated pharmacokinetic parameters, based on the plasma concentrations of itraconazole and hydroxy itraconazole, were AUC0-72h, AUC0-∝, Cmax, Tmax, T1/2 and Kel.Results: The 90% CI for the test/reference geometric means ratio for the parent compound, itraconazole, was in the range from 85.29% to 116.07% for AUC0-72h. Since the coefficient of variation (CV) for the reference product was 44.95% for Cmax, the 90% CI for this parameter for itraconazole was 93.49–133.78%, which was within the proposed limits of the EMA for bioequivalence of 72.15–138.59%. During the study, 4 subjects encountered a total of 14 mild adverse events.Conclusions: The use of the reference-scaling approach with 3-period design (TRR, RTR, and RRT) was an efficient way to demonstrate that two commercially available oral itraconazole formulations met the predetermined bioequivalence criteria.

Population in vitro-in vivo pharmacokinetic model of first-pass metabolism: itraconazole and hydroxy-itraconazole.

The aim of this study was to develop a population in vitro-in vivo pharmacokinetic model that simultaneously describe the absorption and accumulation kinetics of itraconazole (ICZ) and hydroxy-itraconazole (HICZ) in healthy subjects. The model integrated meta-models of gastrointestinal pH and gastrointestinal transit time and in vitro dissolution models of ICZ with the absorption and disposition kinetics of ICZ and HICZ. Mean concentration intravenous data, and single- and multi-dose oral data were used for model development. Model development was conducted in NONMEM in a stepwise manner. First, a model of intravenous data (systemic kinetics) was established and then extended to include the oral data. The latter was then extended to establish the in vitro-in vivo pharmacokinetic model. The systemic disposition of ICZ was best described by a 3-compartment model with oral absorption described by 4-transit compartments and HICZ distribution by a 1-compartment model. ICZ clearance was best described using a mixed inhibition model that allowed HICZ concentrations to inhibit the clearance of parent drug. HICZ clearance was described by Michaelis-Menten elimination kinetics. An in vitro-in vivo model was successfully established for both formulations. The presented model was able to describe ICZ and HICZ plasma concentrations over a wide range of oral and intravenous doses and allowed the exploration of complexities associated with the non-linear ICZ and HICZ kinetics. The model may provide insight into the variability in exposure of ICZ with respect to relating in vivo dissolution characteristics with in vivo disposition kinetics.

Physiologically Based Pharmacokinetic Modeling to Understand the Observed Drug–Drug Interaction of LY2623091 with CYP3A Inhibitors Itraconazole and Diltiazem

Aims: Drug–drug interaction studies were used to estimate CYP3A4-dependent clearance of LY2623091. Methods: in a cross-over design, healthy adults received a single 6-mg dose of LY2623091 at baseline. Itraconazole (200 mg twice on day 1 then daily × 19 days; n = 16) and diltiazem (240 mg Extended Release daily × 13 days; n = 16) were given. On days 6 and 4, respectively, LY2623091 was dosed 1 h after itraconazole/diltiazem. Pharmacokinetic samples were obtained and static and dynamic models were used to assess interaction. Results: Area under the concentration–time curve for LY2623091 increased 2.2-fold with itraconazole and 1.4-fold with diltiazem. Maximum plasma concentration did not change. The physiologically based pharmacokinetic model overpredicted itraconazole and hydroxy–itraconazole concentrations. Extraction by CYP3A4 was ∼0.5; gut wall extraction was negligible. Conclusion: Interaction risk requires early clinical assessment to quantify the candidate drug. (Clinical trial registration number: NCT02300259). Data deposition: deposition pending.

Simultaneous determination of itraconazole, hydroxy itraconazole, keto itraconazole and N-desalkyl itraconazole concentration in human plasma using liquid chromatography with tandem mass spectrometry

A high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS) assay was developed and validated for simultaneous determination of itraconazole (ITZ), hydroxy-itraconazole (OH-ITZ), keto-itraconazole (keto-ITZ) and N-desalkyl itraconazole (ND-ITZ) concentration in human plasma. One hundred and fifty microliters of human plasma were extracted using a solid-supported liquid extraction (SLE) method and the final extracts were analyzed using reverse-phase chromatography and positive electrospray ionization mass spectrometry. The standard curve range is 5-2500 ng/mL for ITZ and OH-ITZ and 0.4-200 ng/mL for keto-ITZ and ND-ITZ. The curve was fitted to a 1/x(2) weighted linear regression model for all analytes. The precision and accuracy of the LC-MS/MS assay based on the five analytical quality control (QC) levels were well within the acceptance criteria from both FDA and EMA guidance for bioanalytical method validation. Average extraction recovery was 97.4% for ITZ, 112.9% for OH-ITZ, 103.4% for keto-ITZ, and 102.3% for ND-ITZ across their respective curve range. Matrix factor was close to 1.0 at both high and low QC levels of all 4 analytes, which indicates minimal ion suppression or enhancement in our validated assay. Itraconazole and all three metabolites are stable in human plasma for 145 days stored at -70 °C freezers. The validated assay was successfully applied to a clinical study, which has a drug-drug interaction (DDI) arm using ITZ as a cytochrome P450, family 3, subfamily A (CYP3A) inhibitor.

Ultra-performance liquid chromatography electrospray ionization–tandem mass spectrometry method for the simultaneous determination of itraconazole and hydroxy itraconazole in human plasma

A highly sensitive, selective, and precise ultra-performance liquid chromatography tandem mass spectrometry method was developed and validated for simultaneous quantification of itraconazole and hydroxy itraconazole in human plasma by a single liquid–liquid extraction step. The precursor to product ion transitions of m/z 705.3/392.3, m/z 721.2/408.3 and m/z 708.2/435.4 were used to detect and quantify itraconazole, hydroxy itraconazole and itraconazole-d3 respectively. The lower limit of quantitation was found to be 0.500ng/mL for itraconazole and 1.00ng/mL for hydroxy itraconazole. The mean recoveries for itraconazole and hydroxy itraconazole were found to be 100.045% and 100.021%, respectively. This developed method with a chromatographic run time of 2.0min was successfully applied to a bioequivalence study of 100mg itraconazole capsule.

A simple, sensitive HPLC-PDA method for the quantification of itraconazole and hydroxy itraconazole in human serum: a reference laboratory experience

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.

Determination of pharmacokinetics of itraconazole in healthy Indian subjects under fed condition and incurred sample analysis using a validated liquid chromatography tandem mass spectrometric method

This study aimed to determine Itraconazole Pharmacokinetics in healthy Indian subjects under fed condition and incurred sample analysis in human plasma by using a validated liquid chromatography tandem mass spectrometric method. A study in healthy human Indian subjects under fed condition was conducted to determine pharmacokinetics of two treatments of Itraconazole 100 mg capsules. Method validation was performed as per USFDA guidelines. Incurred Sample Reanalysis (ISR) was performed by random selection of subject samples. All the method validation parameters were found to be acceptable in terms of accuracy and precision. Results of all stability exercises in human plasma were within acceptable limits. The validated LCMS/MS method was used for sample analysis. A value of 96.7% for Itraconazole and 92.2% for Hydroxy Itraconazole was observed for incurred sample re-analysis. Arithmetic means for Cmax and AUC0→∞ were 103.699, 95.105 ng/ml and 1665.599, 1556.486 ng/ml.h, respectively, for Itraconazole and for...

A comparative bioavailability study of two formulations of itraconazole 100 mg capsule in healthy human Indian subjects under fasting conditions

The objective of the study was to evaluate the effect of time of administration of food post-dosing on the oral bioavailability of two formulations of itraconazole 100 mg capsule in 24 healthy adult Indian subjects under fasting conditions. Three different paradigms for time of administrations of food post-dose were selected: 4 h, 5 h, and 6 h. A randomized two-treatment, four-period, three-sequence, single dose bioavailability study was conducted. After dosing, serial blood samples were collected for up to 72 h. Plasma samples were analyzed for Itraconazole (ITZ) and its metabolite Hydroxy Itraconazole (OH-ITZ) by a sensitive and validated simultaneous liquid-chromatographic and mass-spectrometric (LC-MS/MS) method. A comparison of effect of post-dose fasting up to 4 h, 5 h, and 6 h on pharmacokinetic parameters of the two formulations was done, and it was found that pharmacokinetic parameters (Cmax, AUC0–t, AUC0–∞, Tmax) were consistent for both formulations in all the paradigms tested. This result sugg...

Sensitive Determination of Itraconazole and Its Active Metabolite in Human Plasma by Column-switching High-performance Liquid Chromatography With Ultraviolet Detection

A simple and sensitive column-switching high-performance liquid chromatographic method for the simultaneous determination of itraconazole (ITZ) and its active metabolite, hydroxyitraconazole (HIT) in human plasma is described. ITZ, HIT, and an internal standard, R051012, were extracted from 1 mL of alkalinized plasma sample using n-heptane-chloroform (60:40, vol/vol). The extract was injected onto column I (TSK precolumn BSA-ODS/S, 5 microm, 10 x 4.6 mm ID) for clean-up and column II (Develosil C8-5 column, 5 microm, 150 x 4.6 mm ID) for separation. The mobile phase consisted of phosphate buffer-acetonitrile (68:32 vol/vol, pH 6.0) for clean-up and phosphate buffer-acetonitrile (35:65 vol/vol, pH 6.0) for separation. The peaks were monitored with an ultraviolet detector set at a wavelength of 263 nm, and total time for chromatographic separation was about 24 minutes. The validated concentration ranges of this method were 3 to 500 ng/mL for ITZ and 3 to 1000 ng/mL for HIT. Mean recoveries were 59.7% for ITZ and 72.8% for HIT. Intraday and interday coefficients of variation were less than 4.6% and 5.0% for ITZ, and 4.6% and 4.9% for HIT at the different concentrations. The limit of quantification was 3 ng/mL for both ITZ and HIT. This method was suitable for therapeutic drug monitoring of ITZ and HIT, and was applied to pharmacokinetic studies in human volunteers.

Capillary electrophoresis evidence for the stereoselective metabolism of itraconazole in man.

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.

Determination of itraconazole and hydroxyitraconazole in human serum and plasma by micellar electrokinetic chromatography

The electrokinetic separation of the hydrophobic antimycotic drug itraconazole (ITC) and its major metabolite, hydroxyitraconazole (HITC), by a binary aqueous–organic solvent medium containing sodium dodecylsulfate, by microemulsion electrokinetic chromatography (MEEKC) and by micellar electrokinetic chromatography (MEKC) was studied. The results suggest that the first approach is difficult to apply and that there is no substantial difference between separations performed using MEEKC and MEKC modified with n-butanol. The simpler MEKC method is more than adequate and was thus employed for the analysis of ITC and HITC in human serum and plasma. Separation was achieved in plain fused-silica capillaries having a low-pH buffer (pH 2.2) with sodium dodecyl sulfate micelles and reversed polarity. The addition of 2-propanol and n-butanol enhanced analyte solubility and altered the selectivity of the separation by influencing the magnitude of the electrophoretic component in the separation mechanism. Under optimised conditions and using head-column field-amplified sample stacking, an internal standard, ITC and two forms of HITC could be separated in under 9 min, with detection limits less than 0.01 μg/mL. Analysis of samples from patients currently prescribed ITC revealed a different HITC peak area ratio to that of the standards, suggesting a stereoselective component of ITC metabolisation. Comparison of MEKC data with those of a HPLC method employed on a routine basis showed excellent agreement, indicating the potential of this approach for therapeutic drug monitoring of ITC.

Expedient microdetermination of itraconazole and hydroxyitraconazole in plasma by high-performance liquid chromatography with fluorescence detection.

The authors describe a useful and rapid micromethod for the analysis of itraconazole (ITZ) and its active metabolite hydroxyitraconazole (HIT) in human plasma. After a simple deproteinization of 100 microL plasma with acetonitrile, the drug, its metabolite, and an internal standard (IS, ketoconazole) were separated on an 8 mm x 10 cm NovaPak (Waters Associates; Milford, MA) C(18) 4-microm particle-size radial compression cartridge. The compounds of interest were detected using a fluorescence detector with the excitation wavelength set at 260 nm and the emission at 365 nm. The mobile phase consisted of 420 mL water adjusted to a pH of 2.5 with phosphoric acid, 580 mL acetonitrile, and 100 microL triethylamine, which was delivered at a flow rate of 3.0 mL/min. This expedient and rugged method is being used to monitor therapeutic levels in bone marrow transplant recipients who are taking the drug for prophylaxis.

Discrepancies in bioassay and chromatography determinations explained by metabolism of itraconazole to hydroxyitraconazole: studies of interpatient variations in concentrations

Pharmacologic studies of itraconazole (IZ), a triazole antifungal, indicated unexplained differences between bioassay and chromatographic determinations and large variations in steady-state blood concentrations. We show that concentrations of a hydroxylated metabolite, hydroxyitraconazole (HIZ), are approximately twofold higher than IZ over a range of concentrations. Though HIZ and IZ appear equipotent against selected pathogens, HIZ is two to three times more active against a commonly used bioassay fungus but minimally affects IZ activity. Hence, HIZ probably contributes importantly to the therapeutic activity attributed to IZ and contributes approximately four to six times the activity of IZ in bioassays, explaining discrepancies observed between assay methods.