RESUMEN
AIMS: Objective methods to determine statin adherence are requested to improve lipid management. We have recently established a method to detect reduced adherence to atorvastatin therapy with cut-off values based on the sum of atorvastatin and its major metabolites in the blood. We aimed to validate this method in patients with and without cardiovascular disease, and optimize previous cut-off values. METHODS AND RESULTS: The pharmacokinetic study included 60 participants treated with atorvastatin 20 mg (N = 20), 40 mg (N = 20), and 80 mg (N = 20). Atorvastatin was then stopped and blood samples collected from day zero to day four. Quantification of the parent drug and its metabolites in blood plasma was performed with a liquid chromatography-tandem mass spectrometry assay. The cut-off values for reduced adherence were validated and optimized by calculating diagnostic sensitivity and specificity. Our candidate cut-off value of dose-normalized six-component sum of atorvastatin plus metabolites <0.10 nM/mg provided a sensitivity of 97% and a specificity of 93% for detecting ≥2 omitted doses. An optimized cut-off <0.062 nM/mg provided a sensitivity of 90% and a specificity of 100%. An alternative simplified two-component metabolite sum with a cut-off value <0.05 nM/mg provided a sensitivity of 98% and a specificity of 76%. An optimized cut-off <0.02 nM/mg provided a sensitivity of 97% and a specificity of 98%. CONCLUSION: This validation study confirms that our direct method discriminates reduced adherence from adherence to atorvastatin therapy with high diagnostic accuracy. The method may improve lipid management in clinical practice and serve as a useful tool in future studies.
Asunto(s)
Atorvastatina , Inhibidores de Hidroximetilglutaril-CoA Reductasas , Cumplimiento de la Medicación , Atorvastatina/farmacocinética , Atorvastatina/uso terapéutico , Atorvastatina/sangre , Humanos , Masculino , Femenino , Persona de Mediana Edad , Inhibidores de Hidroximetilglutaril-CoA Reductasas/farmacocinética , Inhibidores de Hidroximetilglutaril-CoA Reductasas/uso terapéutico , Inhibidores de Hidroximetilglutaril-CoA Reductasas/sangre , Inhibidores de Hidroximetilglutaril-CoA Reductasas/administración & dosificación , Anciano , Ácidos Heptanoicos/farmacocinética , Ácidos Heptanoicos/administración & dosificación , Ácidos Heptanoicos/sangre , Ácidos Heptanoicos/uso terapéutico , Pirroles/farmacocinética , Pirroles/sangre , Pirroles/administración & dosificación , Espectrometría de Masas en Tándem , Cromatografía Liquida , Enfermedades Cardiovasculares/tratamiento farmacológico , Enfermedades Cardiovasculares/diagnóstico , Enfermedades Cardiovasculares/sangre , Enfermedades Cardiovasculares/prevención & control , Reproducibilidad de los Resultados , Relación Dosis-Respuesta a DrogaRESUMEN
The antihyerlipidemic drug atorvastatin (ATR) is used worldwide as part of the strategy to prevent cardiovascular events. The high prevalence of patient nonadherence remains an important challenge which could be addressed efficiently by precision pharmacotherapy based on therapeutic drug monitoring (TDM). ATR is metabolized to pharmacologically active metabolites, and evidence shows that the sums of ATR acid and lactone form concentrations (ATR + ATRL), or of ATR and hydroxylated metabolites (ATR + MET) should be assayed. A method is presented for the analysis of these substances in serum. Method validation included the estimation of the quantitative relationship between the concentrations and the standard deviations (SD), which supports the optimal incorporation of TDM results into nonparametric pharmacokinetic models. The concentrations of the analytes were evaluated in human subjects receiving ATR. The method's performance improved by taking the sums of acid and lactone concentrations into account. The concentration-SD relationship was linear, and we recommend applying Theil's regression for estimating the assay error. All analytes could be detected by 2 h post dose in the samples of human subjects. The changes in metabolite/parent drug concentration ratios in time depended on the dose. The method is suitable for the TDM of ATR with a focus on precision pharmacotherapy.
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Atorvastatina/sangre , Cromatografía Liquida/métodos , Monitoreo de Drogas/métodos , Ácidos Heptanoicos/sangre , Lactonas/sangre , Medicina de Precisión , Espectrometría de Masas en Tándem/métodos , HumanosRESUMEN
A systematic methodology was used to quantify ganoderic acid-A (GA-A) loaded nano-lipid carriers (NLC) in rat plasma using UPLC-MS/MS. Separation of the analyte was achieved using ACQUITY UPLC BEH C18 column (1.7 µm) and mobile phase as water containing 0.1% Acetonitrile (40: 60% v/v) at a flow rate of 0.4 mL·min-1. The analyte was detected using MRM mode to track precursor-to-product ion transitions of 515.37 â 285.31 m/z (time scan of 2 min) for GA-A, and 175.11 â 115.08 m/z (time scan of 4 min) for ascorbic acid as an internal standard (IS), respectively. The developed method was validated for linearity, accuracy, within and between day precisions, limit of quantification and recovery of the analyte. The results indicated intra and inter-day consistency and precision values were found to be within the acceptance limit for the plasma samples. The method applicability for determination of pharmacokinetic parameters of GA-A was assessed after oral administration of free GA-A solution and GA-A-loaded NLC, which indicated significant difference (p < 0.05) in the rate and extent of absorption parameters of GA-A from the NLC formulation vis-à-vis the plain solution. Overall, the studies construed successful development and application of UPLC-MS/MS method for estimation of GA-A in the lipidic formulation.
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Ácidos Heptanoicos/sangre , Ácidos Heptanoicos/farmacocinética , Lanosterol/análogos & derivados , Espectrometría de Masas en Tándem/métodos , Animales , Portadores de Fármacos/química , Ácidos Heptanoicos/química , Lanosterol/sangre , Lanosterol/química , Lanosterol/farmacocinética , Límite de Detección , Modelos Lineales , Lípidos/química , Masculino , Nanopartículas/química , Ratas , Ratas Wistar , Reproducibilidad de los ResultadosRESUMEN
Short-chain fatty acids, the end products of fermentation of dietary fibers by the gut microbiota, have been shown to exert multiple effects on mammalian metabolism. For the analysis of short-chain fatty acids, gas chromatography-mass spectrometry is a very powerful and reliable method. Here, we describe a fast, reliable, and reproducible method for the separation and quantification of short-chain fatty acids in mouse feces, cecum content, and blood samples (i.e., plasma or serum) using gas chromatography-mass spectrometry. The short-chain fatty acids analyzed include acetic acid, propionic acid, butyric acid, valeric acid, hexanoic acid, and heptanoic acid.
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Ciego/química , Ácidos Grasos Volátiles/análisis , Heces/química , Metabolómica/métodos , Ácido Acético/análisis , Ácido Acético/sangre , Animales , Ácido Butírico/análisis , Ácido Butírico/sangre , Caproatos/análisis , Caproatos/sangre , Ácidos Grasos Volátiles/sangre , Cromatografía de Gases y Espectrometría de Masas , Ácidos Heptanoicos/análisis , Ácidos Heptanoicos/sangre , Ratones , Ácidos Pentanoicos/análisis , Ácidos Pentanoicos/sangre , Propionatos/análisis , Propionatos/sangre , Reproducibilidad de los ResultadosRESUMEN
We examined associations between prenatal exposure to perfluorohexane sulfonic acid (PFHxS), perfluoroheptanoic acid (PFHpA), perfluorononanoic acid (PFNA), and perfluorodecanic acid (PFDA) - and child behaviour (SDQ-total) and hyperactivity (sub-scale) at 5-9years of age in birth cohorts from Greenland and Ukraine. Pregnancy serum samples (N=1023) were analysed for perfluoroalkyl substances (PFASs) and categorised into tertiles and also used as continuous exposure variables. Problem behaviour and hyperactivity were assessed, using the Strength and Difficulties Questionnaire (SDQ) and categorised as normal/borderline and abnormal. Associations were analysed using multiple logistic and linear regression. High compared to low prenatal PFHxS exposure was associated with 1.16 (95% confidence interval (CI): 0.08; 2.25) point higher SDQ-total (more problem behaviour) in Greenland and 0.80 (CI: 0.06; 1.54) point higher SDQ-total in the combined analyses, whereas no association was present in Ukraine alone. One natural log-unit increase in prenatal PFNA exposure was associated with 0.90 (CI: 0.10; 1.71) points higher SDQ-total in Greenland and 0.72 (CI: 0.13; 1.31) points higher in the combined analysis and no association in Ukraine. Prenatal PFAS exposure was unrelated to problem behaviour (abnormal SDQ-total). In the combined analysis, odds ratio (OR) (CI) for hyperactivity was 1.8 (1.0; 3.2) for one natural log-unit increase in prenatal PFNA and 1.7 (1.0; 3.1) for one natural log-unit increase in prenatal PFDA exposure. Findings are compatible with weak effects on child behaviour of prenatal exposure to some PFASs although spurious results are not entirely unlikely. The associations were strongest in Greenland.
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Ácidos Alcanesulfónicos/toxicidad , Conducta Infantil/efectos de los fármacos , Desarrollo Infantil/efectos de los fármacos , Fluorocarburos/toxicidad , Efectos Tardíos de la Exposición Prenatal , Adulto , Ácidos Alcanesulfónicos/sangre , Análisis Químico de la Sangre , Niño , Preescolar , Femenino , Fluorocarburos/sangre , Ácidos Heptanoicos/sangre , Ácidos Heptanoicos/toxicidad , Humanos , Estudios Longitudinales , Masculino , Embarazo , Efectos Tardíos de la Exposición Prenatal/sangre , Efectos Tardíos de la Exposición Prenatal/inducido químicamente , Efectos Tardíos de la Exposición Prenatal/fisiopatología , Encuestas y Cuestionarios , Adulto JovenRESUMEN
Ganoderic acid A (GAA), an active triterpenoid of the traditional Chinese herbal medicine Lingzhi, has been reported to exhibit antinociceptive, antioxidative, and anti-cancer activities. The present study aims to establish a sensitive and rapid UPLC-MS/MS method for studying the plasma and brain pharmacokinetics of GAA in rats. The analytes were separated on a C18 column eluted with a gradient mobile phase consisting of acetonitrile and 0.1% aqueous formic acid at 0.3mL/min. The eluate was monitored by a mass detector using an MRM (m/z, 515.3-285.1) model in negative electrospray ionization. The calibration curve showed good linearity (r2>0.99), with limits of detection and quantification of 0.25 and 2.00 nmol/L, respectively. The intra- and inter-day precision and accuracy were less than 9.99% and ranged from 97.45% to 114.62%, respectively. The extraction recovery from plasma was between 92.89% and 98.87%. GAA was found to be stable in treated samples at room temperature (22°C) for 12h and in plasma at -20°C for 7d. The developed method was successfully applied to a pharmacokinetic study of GAA in rats. GAA could be rapidly absorbed into the circulation (Tmax, 0.15h) and eliminated relatively slowly (t1/2, 2.46h) after orally dosing, and could also be detected in the brain lateral ventricle (Tmax, 0.25h and t1/2, 1.40h) after intravenously dosing. The absolute oral bioavailability and brain permeability of GAA were estimated to be 8.68% and 2.96%, respectively.
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Encéfalo/metabolismo , Cromatografía Líquida de Alta Presión/métodos , Medicamentos Herbarios Chinos/farmacocinética , Ácidos Heptanoicos/sangre , Ácidos Heptanoicos/líquido cefalorraquídeo , Lanosterol/análogos & derivados , Espectrometría de Masas en Tándem/métodos , Analgésicos/sangre , Analgésicos/líquido cefalorraquídeo , Animales , Antineoplásicos Fitogénicos/sangre , Antineoplásicos Fitogénicos/líquido cefalorraquídeo , Antioxidantes/farmacocinética , Lanosterol/sangre , Lanosterol/líquido cefalorraquídeo , Límite de Detección , Masculino , Microdiálisis/métodos , Ratas Sprague-DawleyRESUMEN
Millions of individuals are treated with a variety of statins that are metabolized to a variety of active metabolites. A single assay capable of simultaneously quantifying commonly used statins and their major metabolites has not been previously reported. Herein we describe the development and validation of a novel and robust liquid chromatography-tandem mass spectrometry assay for simultaneously quantifying simvastatin, lovastatin, atorvastatin, and their metabolites, simvastatin acid, lovastatin acid, para-hydroxy atorvastatin, and ortho-hydroxy atorvastatin in human plasma. Plasma samples were processed with a simple protein precipitation technique using acetonitrile, followed by chromatographic separation using an Agilent Zorbax Extend C18 column. A 12.0min linear gradient elution was used at a flow rate of 400µL/min with a mobile phase of water and methanol, both modified with 2mM ammonium formate and 0.2% formic acid. The analytes and internal standard, hesperetin, were detected using the selected reaction monitoring mode on a TSQ Quantum Discovery mass spectrometer with positive electrospray ionization. The assay exhibited a linear range of 1-1000nM for simvastatin acid and lovastatin acid, and a linear range of 0.1-100nM for the other analytes in human plasma. The accuracy and the within- and between-day precisions of the assay were within acceptable ranges, and the method was successfully utilized to quantify the statins and their metabolites in human plasma samples collected from an ongoing pharmacokinetic study.
Asunto(s)
Cromatografía Liquida/métodos , Ácidos Heptanoicos/sangre , Lovastatina/sangre , Pirroles/sangre , Simvastatina/análogos & derivados , Espectrometría de Masas en Tándem/métodos , Adulto , Anciano , Atorvastatina , Calibración , Ácidos Heptanoicos/química , Humanos , Lovastatina/química , Persona de Mediana Edad , Pirroles/química , Control de Calidad , Reproducibilidad de los Resultados , Simvastatina/sangre , Simvastatina/químicaRESUMEN
OBJECTIVE: To assess California firefighters' blood concentrations of selected chemicals and compare with a representative US population. METHODS: We report laboratory methods and analytic results for cadmium, lead, mercury, and manganese in whole blood and 12 serum perfluorinated chemicals in a sample of 101 Southern California firefighters. RESULTS: Firefighters' blood metal concentrations were all similar to or lower than the National Health and Nutrition Examination Survey (NHANES) values, except for six participants whose mercury concentrations (range: 9.79 to 13.42 µg/L) were close to or higher than the NHANES reporting threshold of 10 µg/L. Perfluorodecanoic acid concentrations were elevated compared with NHANES and other firefighter studies. CONCLUSIONS: Perfluorodecanoic acid concentrations were three times higher in this firefighter group than in NHANES adult males. Firefighters may have unidentified sources of occupational exposure to perfluorinated chemicals.
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Monitoreo del Ambiente/métodos , Bomberos , Incendios , Exposición Profesional/análisis , Adulto , Factores de Edad , Ácidos Alcanesulfónicos/sangre , Cadmio/sangre , California , Ácidos Decanoicos/sangre , Ácidos Grasos , Femenino , Fluorocarburos/sangre , Ácidos Heptanoicos/sangre , Humanos , Hidrocarburos Fluorados/sangre , Plomo/sangre , Masculino , Manganeso/sangre , Mercurio/sangre , Persona de Mediana Edad , Encuestas Nutricionales , Exposición Profesional/normas , Sulfonamidas/sangre , Adulto JovenRESUMEN
A novel and sensitive dispersive liquid-liquid microextraction method based on the solidification of the floating organic drop combined with high-performance liquid chromatography and ultraviolet detection was used for the determination of atorvastatine in blood serum samples. The chromatographic separation of atorvastatin was carried out using methanol as the mobile phase organic modifier. Various parameters affecting the extraction efficiency were optimized, such as the kind and volume of extraction solvent (1-undecanol) and disperser solvent (acetonitrile), pH, and the extraction time. The calibration curve was linear in the range of 0.2-6000 µg/L of atorvastatin (r(2) = 0.995) with a limit of detection of 0.07 µg/L. The relative standard deviation for 100 µg/L of atorvastatin in human plasma was 8.4% (n = 4). The recoveries of plasma samples spiked with atorvastatin were in the range of 98.8-113.8%. The obtained results showed that the proposed method is fast, simple, and reliable for the determination of very low concentrations of atorvastatin in human plasma samples.
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Cromatografía Líquida de Alta Presión/métodos , Ácidos Heptanoicos/sangre , Inhibidores de Hidroximetilglutaril-CoA Reductasas/sangre , Microextracción en Fase Líquida/métodos , Pirroles/sangre , Adulto , Atorvastatina , Femenino , Humanos , Concentración de Iones de Hidrógeno , Límite de Detección , Masculino , Cloruro de Sodio/química , Espectrofotometría Ultravioleta , Adulto JovenRESUMEN
The aim of the present study was to investigate the effect of Soluplus® on the solubility of atorvastatin calcium and to develop a solid dispersion formulation that can improve the oral bioavailability of atorvastatin calcium. We demonstrated that Soluplus® increases the aqueous solubility of atorvastatin calcium. Several solid dispersion formulations of atorvastatin calcium with Soluplus® were prepared at various drug : carrier ratios by spray drying. Physicochemical analysis demonstrated that atorvastatin calcium is amorphous in each solid dispersion, and the 2 : 8 drug : carrier ratio provided the highest degree of sustained atorvastatin supersaturation. Pharmacokinetic analysis in rats revealed that the 2 : 8 dispersion significantly improved the oral bioavailability of atorvastatin. This study demonstrates that spray-dried Soluplus® solid dispersions can be an effective method for achieving higher atorvastatin plasma levels.
Asunto(s)
Desecación , Ácidos Heptanoicos/química , Ácidos Heptanoicos/farmacocinética , Polietilenglicoles/química , Polietilenglicoles/farmacología , Polivinilos/química , Polivinilos/farmacología , Pirroles/química , Pirroles/farmacocinética , Administración Oral , Animales , Atorvastatina , Disponibilidad Biológica , Ácidos Heptanoicos/administración & dosificación , Ácidos Heptanoicos/sangre , Masculino , Pirroles/administración & dosificación , Pirroles/sangre , Ratas , Ratas Sprague-Dawley , Solubilidad/efectos de los fármacos , Agua/químicaRESUMEN
The combination of ultrahigh-resolution mass spectrometry imaging (UHRMSI) and ultrahigh-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC/MS/MS) was used for the identification and the spatial localization of atorvastatin (AT) and its metabolites in rat tissues. Ultrahigh-resolution and high mass accuracy measurements on a matrix-assisted laser desorption/ionization (MALDI)-Orbitrap mass spectrometer allowed better detection of desired analytes in the background of matrix and endogenous compounds. Tandem mass spectra were also used to confirm the identification of detected metabolites in complex matrices. The optimization of sample preparation before imaging experiments included the tissue cryogenic sectioning (thickness 20 µm), the transfer to stainless steel or glass slide, and the selection of suitable matrix and its homogenous deposition on the tissue slice. Thirteen matrices typically used for small molecule analysis, e.g., 2,5-dihydroxybenzoic acid (DHB), 1,5-diaminonaphthalene (DAN), 9-aminoacridine (AA), etc., were investigated for the studied drug and its metabolite detection efficiency in both polarity modes. Particular matrices were scored based on the strength of extracted ion current (EIC), relative ratio of AT molecular adducts, and fragment ions. The matrix deposition on the tissue for the most suitable matrices was done by sublimation to obtain the small crystal size and to avoid local variations in the ionization efficiency. UHPLC/MS profiling of drug metabolites in adjacent tissue slices with the previously optimized extraction was performed in parallel to mass spectrometry imaging (MSI) measurements to obtain more detailed information on metabolites in addition to the spatial information from MSI. The quantitation of atorvastatin in rat liver, serum, and feces was also performed.
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Cromatografía Líquida de Alta Presión/métodos , Heces/química , Ácidos Heptanoicos/metabolismo , Hígado/química , Pirroles/metabolismo , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Espectrometría de Masas en Tándem/métodos , Animales , Atorvastatina , Ácidos Heptanoicos/sangre , Masculino , Pirroles/sangre , Ratas , Ratas Wistar , Distribución Tisular/fisiologíaRESUMEN
PURPOSE: ACT-178882, a direct renin inhibitor, was used as a model compound in an elaborate drug-drug interaction study with atorvastatin and simvastatin to explore complex CYP3A4 inductive and inhibitory properties. METHODS: Thirty-two healthy male subjects received single doses of 20 mg atorvastatin and 20 mg simvastatin on days 1, 9, 31, and 41. On days 6 to 33, 500 mg ACT-178882 was administered once daily. Plasma concentrations of ACT-178882, simvastatin, and atorvastatin were measured by LC-MS/MS. Routine safety assessments were performed throughout the study. RESULTS: Exposure (as based on area under the curve) to simvastatin and 6ß-hydroxyacid simvastatin increased (90 % confidence interval) 4.63-fold (3.90, 5.50) and 3.71-fold (3.19, 4.32), respectively, when comparing day 9 and day 1. On day 9, exposure to atorvastatin was similar but Cmax decreased, while both variables decreased for ortho-hydroxy atorvastatin when compared to day 1. On day 31, after prolonged administration of ACT-178882, exposure to atorvastatin, ortho-hydroxy atorvastatin, simvastatin, and 6ß-hydroxyacid simvastatin decreased by 14, 19, 21, and 27 %, respectively, when compared to day 9. However, on this day, exposure to simvastatin and its metabolite was still markedly higher when compared to day 1. Effects of ACT-178882 had largely dissipated on day 41. CONCLUSIONS: This design enabled the study of complex time-dependent effects on CYP3A4 activity with clinically relevant substrates.
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Ciclopropanos/farmacología , Inductores del Citocromo P-450 CYP3A/farmacología , Inhibidores del Citocromo P-450 CYP3A/farmacología , Citocromo P-450 CYP3A/metabolismo , Inhibidores de Hidroximetilglutaril-CoA Reductasas/farmacocinética , Piridinas/farmacología , Adolescente , Adulto , Atorvastatina , Ciclopropanos/administración & dosificación , Ciclopropanos/efectos adversos , Ciclopropanos/sangre , Citocromo P-450 CYP3A/biosíntesis , Inductores del Citocromo P-450 CYP3A/administración & dosificación , Inductores del Citocromo P-450 CYP3A/efectos adversos , Inductores del Citocromo P-450 CYP3A/sangre , Inhibidores del Citocromo P-450 CYP3A/administración & dosificación , Inhibidores del Citocromo P-450 CYP3A/efectos adversos , Inhibidores del Citocromo P-450 CYP3A/sangre , Interacciones Farmacológicas , Ácidos Heptanoicos/administración & dosificación , Ácidos Heptanoicos/sangre , Ácidos Heptanoicos/farmacocinética , Humanos , Inhibidores de Hidroximetilglutaril-CoA Reductasas/administración & dosificación , Inhibidores de Hidroximetilglutaril-CoA Reductasas/sangre , Masculino , Persona de Mediana Edad , Piridinas/administración & dosificación , Piridinas/efectos adversos , Piridinas/sangre , Pirroles/administración & dosificación , Pirroles/sangre , Pirroles/farmacocinética , Simvastatina/administración & dosificación , Simvastatina/sangre , Simvastatina/farmacocinética , Especificidad por Sustrato , Adulto JovenRESUMEN
Atorvastatin is eliminated by CYP3A4 which follows carrier-mediated uptake into hepatocytes by OATP1B1, OATP1B3, and OATP2B1. Multiple clinical studies demonstrated that OATP inhibition by rifampin had a greater impact on atorvastatin systemic concentration than itraconazole-mediated CYP3A4 inhibition. If it is assumed that the blood and hepatocyte compartments are differentiated by the concentration gradient that is established by OATPs, and if the rate of uptake into the hepatocyte is rate-determining to the elimination of atorvastatin from the body, then it is hypothesized that blood concentrations may not necessarily reflect liver concentrations. In wild-type mice, rifampin had a greater effect on systemic exposure of atorvastatin than ketoconazole, as the blood area under the blood concentration-time curve increased 7- and 2-fold, respectively. In contrast, liver concentrations were affected more by ketoconazole than by rifampin, as liver levels increased 21- and 4-fold, respectively. Similarly, in Cyp3a knockout animals, 39-fold increases in liver concentrations were observed despite insignificant changes in the blood area under the blood concentration-time curve. Interestingly, blood and liver levels in Oatp1a/b knockout animals were similar to wild types, suggesting that Oatp1a/b knockout may be necessary but not sufficient to completely describe atorvastatin uptake in mice. Data presented in this work indicate that there is a substantial drug interaction when blocking atorvastatin metabolism, but the effects of this interaction are predominantly manifested in the liver and may not be captured when monitoring changes in the systemic circulation. Consequently, there may be a disconnect when trying to relate blood exposure to instances of hepatotoxicity because a pharmacokinetic-toxicity relationship may not be obvious from blood concentrations.
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Sistema Enzimático del Citocromo P-450/deficiencia , Ácidos Heptanoicos/sangre , Cetoconazol/farmacocinética , Hígado/metabolismo , Proteína 1 de Transporte de Anión Orgánico/deficiencia , Pirroles/sangre , Rifampin/farmacocinética , Animales , Atorvastatina , Citocromo P-450 CYP3A , Interacciones Farmacológicas/fisiología , Femenino , Células HEK293 , Ácidos Heptanoicos/metabolismo , Humanos , Hígado/efectos de los fármacos , Ratones , Ratones Noqueados , Pirroles/metabolismoRESUMEN
Atorvastatin and ezetimibe are lipid-lowering drugs prescribed for the treatment of hypercholesterolemia. An LC-MS-MS method has been developed and validated for the simultaneous estimation of atorvastatin and ezetimibe in human plasma using pitavastatin as an internal standard. Liquid-liquid extraction was used for the purification and preconcentration of analytes from human plasma matrix. The chromatographic separation was achieved within 3.0 min by an isocratic mobile phase consisting of 0.2% formic acid in water-acetonitrile (30:70, v/v), flowing through Agilent Eclipse-plus C18, 100 × 4.6 mm, 3.5 µm analytical column, at a flow rate of 0.6 mL min(-1). Multiple reaction monitoring transitions were measured in the positive ion mode for atorvastatin and internal standard, while ezetimibe was measured in negative ion mode. A detailed validation of the method was performed as per US-FDA guidelines and the standard curves were found to be linear in the range of 0.2-30.0 ng mL(-1) with a mean correlation coefficient >0.999 for both drugs. In human plasma, atorvastatin and ezetimibe were stable for at least 36 days at -70 ± 5 °C and 6 h at ambient temperature. After extraction from plasma, the reconstituted samples of atorvastatin and ezetimibe were stable in an autosampler at ambient temperature for 6 h. Also, the cited drugs were stable in plasma samples upon subjecting to three freeze thaw cycles. The method is simple, specific, sensitive, precise, accurate and suitable for bioequivalence and pharmacokinetic studies of this combination.
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Azetidinas/sangre , Cromatografía Liquida/métodos , Ácidos Heptanoicos/sangre , Pirroles/sangre , Espectrometría de Masas en Tándem/métodos , Atorvastatina , Ezetimiba , Humanos , Reproducibilidad de los ResultadosRESUMEN
The present study aimed to evaluate the impact of CYP3A4*1G allele on the pharmacokinetics of atorvastatin in the Chinese Han patients with coronary heart disease (CHD). Twenty male patients of CHD with different CYP3A4*1G genotypes were orally administered a single 20 mg dose of atorvastatin. Plasma concentrations of atorvastatin and 2-hydroxyatorvastatin were measured by high-performance liquid chromatography tandem mass spectrometry. The mean area under the plasma concentration-time curve from 0 to infinity (AUC0-∞ ) of atorvastatin in subjects with the CYP3A4*1G/*1G genotype were 36% or 25% lower than in those with the wild-type or the *1/*1G genotype, respectively. The time to peak plasma concentration (Tmax ) and oral clearance of atorvastatin (CL/F) were significantly different between subjects with the CYP3A4*1G/*1G genotype and the wild-type. The AUC0-∞ for 2-hydroxyatorvastatin in subjects with the CYP3A4*1G/*1G genotype was 44% or 31% lower than in those with the wild-type or the *1/*1G genotype, respectively. The peak plasma concentration, Tmax and apparent clearance of 2-hydroxyatorvastatin (CL/Fm) were significantly different between subjects with the CYP3A4*1G/*1G genotype and the wild-type. This study indicates that the CYP3A4*1G allele is associated with the pharmacokinetics of atorvastatin and its metabolites in those Chinese Han patients with CHD after a single oral dose.
Asunto(s)
Enfermedad Coronaria/genética , Citocromo P-450 CYP3A/genética , Ácidos Heptanoicos/farmacocinética , Inhibidores de Hidroximetilglutaril-CoA Reductasas/farmacocinética , Pirroles/farmacocinética , Alelos , Pueblo Asiatico/genética , Atorvastatina , Enfermedad Coronaria/metabolismo , Genotipo , Ácidos Heptanoicos/sangre , Humanos , Inhibidores de Hidroximetilglutaril-CoA Reductasas/sangre , Masculino , Persona de Mediana Edad , Pirroles/sangreRESUMEN
PURPOSE: Atorvastatin calcium (ATC) is classified as class II (low solubility and high permeability) compound according to the biopharmaceutical classification system. The amorphous form of ATC possesses higher solubility, dissolution rate, and bioavailability than its crystalline form. Coamorphous drug system is a new and emerging method to prepare stable amorphous forms, in this case leading to the improved stability of ATC in dissolution medium. METHODS: In this study, coamorphous form of ATC and nicotinamide (ATC-NIC) was prepared from solvent evaporation method and characterized using differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FT-IR) and powder X-ray diffraction (PXRD). The intrinsic dissolution rate and solubility of ATC-NIC were determined along with plasma concentrations of ATC using HPLC after oral dosing in rats. RESULTS: The crystalline ATC was converted to coamorphous form revealing a molecular interaction between ATC and NIC. The intrinsic dissolution rate, solubility and plasma concentration of coamorphous ATC-NIC are higher than those of crystalline ATC. ATC-NIC coamorphous system showed greater solution stability than those reported in the literature for amorphous ATC. CONCLUSIONS: Coamorphous ATC-NIC has improved physicochemical and pharmacokinetic properties as compared to ATC. This article is open to POST-PUBLICATION REVIEW. Registered readers (see "For Readers") may comment by clicking on ABSTRACT on the issue's contents page.
Asunto(s)
Ácidos Heptanoicos/farmacocinética , Inhibidores de Hidroximetilglutaril-CoA Reductasas/farmacocinética , Niacinamida/farmacocinética , Pirroles/farmacocinética , Animales , Atorvastatina , Femenino , Ácidos Heptanoicos/sangre , Ácidos Heptanoicos/química , Inhibidores de Hidroximetilglutaril-CoA Reductasas/sangre , Inhibidores de Hidroximetilglutaril-CoA Reductasas/química , Niacinamida/sangre , Niacinamida/química , Pirroles/sangre , Pirroles/química , Ratas , Ratas Wistar , SolubilidadRESUMEN
BACKGROUND: A barrier to statin therapy is myopathy associated with elevated systemic drug exposure. Our objective was to examine the association between clinical and pharmacogenetic variables and statin concentrations in patients. METHODS AND RESULTS: In total, 299 patients taking atorvastatin or rosuvastatin were prospectively recruited at an outpatient referral center. The contribution of clinical variables and transporter gene polymorphisms to statin concentration was assessed using multiple linear regression. We observed 45-fold variation in statin concentration among patients taking the same dose. After adjustment for sex, age, body mass index, ethnicity, dose, and time from last dose, SLCO1B1 c.521T>C (P<0.001) and ABCG2 c.421C>A (P<0.01) were important to rosuvastatin concentration (adjusted R(2)=0.56 for the final model). Atorvastatin concentration was associated with SLCO1B1 c.388A>G (P<0.01) and c.521T>C (P<0.05) and 4ß-hydroxycholesterol, a CYP3A activity marker (adjusted R(2)=0.47). A second cohort of 579 patients from primary and specialty care databases were retrospectively genotyped. In this cohort, genotypes associated with statin concentration were not differently distributed among dosing groups, implying providers had not yet optimized each patient's risk-benefit ratio. Nearly 50% of patients in routine practice taking the highest doses were predicted to have statin concentrations greater than the 90th percentile. CONCLUSIONS: Interindividual variability in statin exposure in patients is associated with uptake and efflux transporter polymorphisms. An algorithm incorporating genomic and clinical variables to avoid high atorvastatin and rosuvastatin levels is described; further study will determine whether this approach reduces incidence of statin myopathy.
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Fluorobencenos/uso terapéutico , Ácidos Heptanoicos/uso terapéutico , Inhibidores de Hidroximetilglutaril-CoA Reductasas/uso terapéutico , Hipercolesterolemia/tratamiento farmacológico , Pirimidinas/uso terapéutico , Pirroles/uso terapéutico , Sulfonamidas/uso terapéutico , Transportador de Casetes de Unión a ATP, Subfamilia G, Miembro 2 , Transportadoras de Casetes de Unión a ATP/genética , Adulto , Anciano , Anciano de 80 o más Años , Alelos , Atorvastatina , Colesterol/sangre , Estudios de Cohortes , Citocromo P-450 CYP3A/genética , Bases de Datos Factuales , Femenino , Fluorobencenos/sangre , Fluorobencenos/farmacocinética , Genotipo , Ácidos Heptanoicos/sangre , Ácidos Heptanoicos/farmacocinética , Humanos , Hidroxicolesteroles/sangre , Modelos Lineales , Transportador 1 de Anión Orgánico Específico del Hígado , Masculino , Persona de Mediana Edad , Proteínas de Neoplasias/genética , Transportadores de Anión Orgánico/genética , Polimorfismo Genético , Polimorfismo de Nucleótido Simple , Estudios Prospectivos , Pirimidinas/sangre , Pirimidinas/farmacocinética , Pirroles/sangre , Pirroles/farmacocinética , Estudios Retrospectivos , Rosuvastatina Cálcica , Sulfonamidas/sangre , Sulfonamidas/farmacocinéticaRESUMEN
1. 5-(N-(4-((4-ethylbenzyl)thio)phenyl)sulfamoyl)-2-methyl benzoic acid (CP-778875), an agonist of the peroxisome proliferator-activated receptor alpha, has been evaluated in the clinic to treat dyslipidemia and type 2 diabetes mellitus. Herein, we investigate the effect of CP-778875 on the pharmacokinetics of atorvastatin acid and its metabolites in humans. 2. The study incorporated a fixed-sequence design conducted in two groups. Group A was designed to estimate the effects of multiple doses of CP-778875 on the single dose pharmacokinetics of atorvastatin. Subjects in group A (n = 26) received atorvastatin (40 mg) on days 1 and 9 and CP-778875 (1.0 mg QD) on days 5-12. Group B was designed to examine the effects of multiple doses of atorvastatin on the single dose pharmacokinetics of CP-778875. Subjects in group B (n = 29) received CP-778875 (0.3 mg) on days 1 and 9 and atorvastatin (40 mg QD) on days 5-12. 3. Mean maximum serum concentration (Cmax) and area under the curve of atorvastatin were increased by 45% and 20%, respectively, upon co-administration with CP-778875. Statistically significant increases in the systemic exposure of ortho- and para-hydroxyatorvastatin were also observed upon concomitant dosing with CP-778875. CP-778875 pharmacokinetics, however, were not impacted upon concomitant dosing with atorvastatin. 4. Inhibition of organic anion transporting polypeptide 1B1 by CP-778875 (IC50 = 2.14 ± 0.40 µM) could be the dominant cause of the pharmacokinetic interaction as CP-778875 did not exhibit significant inhibition of cytochrome P450 3A4/3A5, multidrug resistant protein 1 or breast cancer resistant protein, which are also involved in the hepatobiliary disposition of atorvastatin.
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Benzoatos/farmacología , Ácido Benzoico/farmacología , Ácidos Heptanoicos/farmacología , PPAR alfa/agonistas , Pirroles/farmacología , Sulfonamidas/farmacología , Animales , Atorvastatina , Benzoatos/química , Ácido Benzoico/química , Inhibidores Enzimáticos del Citocromo P-450 , Sistema Enzimático del Citocromo P-450/metabolismo , Perros , Interacciones Farmacológicas , Células HEK293 , Ácidos Heptanoicos/sangre , Ácidos Heptanoicos/farmacocinética , Humanos , Hidroxilación/efectos de los fármacos , Células de Riñón Canino Madin Darby , Proteínas de Transporte de Membrana/metabolismo , Oxidación-Reducción/efectos de los fármacos , Pirroles/sangre , Pirroles/farmacocinética , Sulfonamidas/química , Factores de TiempoRESUMEN
BACKGROUND: Atorvastatin is commonly used to reduce cholesterol. Atorvastatin acid is converted to its corresponding lactone form spontaneously or via glucuronidation mediated by uridine diphosphate glucuronosyltransferase (UGT) 1A1 and 1A3. Atorvastatin lactone is pharmacologically inactive, but is suspected to be muscle toxic and cause statin-induced myopathy (SIM). A several fold increase in systemic exposure of atorvastatin lactone has previously been observed in patients with SIM compared with healthy control subjects. In this study we aimed to investigate the association between polymorphisms in the UGT1A gene locus and plasma atorvastatin lactone levels. METHODS: DNA was extracted from whole blood obtained from a previous pharmacokinetic study of patients carefully diagnosed as having true SIM (n = 13) and healthy control subjects (n = 15). The UGT1A1*28(TA) 7 , UGT1A3*2, UGT1A3*3, and UGT1A3*6 polymorphisms were detected by pyrosequencing. RESULTS: Carriers of the low-expression allele UGT1A1*28(TA) 7 tended to have lower levels of atorvastatin lactone (p < 0.05) than carriers with the normal-activity allele (TA) 6 . CONCLUSION: The low-expression UGT1A1*28(TA) 7 allele seems to be associated with decreased systemic exposure of the suspected muscle-toxic metabolite atorvastatin lactone.
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Glucuronosiltransferasa/genética , Ácidos Heptanoicos/efectos adversos , Lactonas/efectos adversos , Enfermedades Musculares/genética , Pirroles/efectos adversos , Adulto , Alelos , Atorvastatina , Femenino , Genotipo , Ácidos Heptanoicos/administración & dosificación , Ácidos Heptanoicos/sangre , Humanos , Lactonas/administración & dosificación , Lactonas/sangre , Masculino , Persona de Mediana Edad , Enfermedades Musculares/inducido químicamente , Enfermedades Musculares/patología , Polimorfismo Genético , Pirroles/administración & dosificación , Pirroles/sangreRESUMEN
In this paper, we describe a new combination method based on polytetrafluorethylene (PTFE) film-based liquid three-phase micro extraction coupled with differential pulse voltammetry (DPV) for the micro extraction and quantification of atorvastatin calcium (ATC) at the ultra-trace level. Different factors affecting the liquid-three phases micro extraction of atorvastatin calcium, including organic solvent, pH of the donor and acceptor phases, concentration of salt, extraction time, stirring rate and electrochemical factors, were investigated, and the optimal extraction conditions were established. The final stable signal was achieved after a 50 min extraction time, which was used for analytical applications. An enrichment factor of 21 was achieved, and the relative standard deviation (RSD) of the method was 4.5% (n = 4). Differential pulse voltammetry exhibited two wide linear dynamic ranges of 20.0-1000.0 pmol L(-1) and 0.001-11.0 µmol L(-1) of ATC. The detection limit was found to be 8.1 pmol L(-1) ATC. Finally, the proposed method was used as a new combination method for the determination of atorvastatin calcium in real samples, such as human urine and plasma.