RESUMEN

Mizolastine is an antihistamine drug that is commonly used for treatment of chronic urticaria and allergic rhinitis. In this study, a facile, rapid, and sustainable fluorimetric method was established for the estimation of mizolastine in pharmaceutical and biological matrices for the first time. The approach methodology relied on the direct assessment of mizolastine's intrinsic fluorescence at 313 nm after excitation at 272 nm. This intrinsic fluorescence, stemming from the benzimidazole fluorophore moiety in mizolastine structure, serves as a distinctive marker for its precise quantification in the spiked human plasma and pharmaceutical formulations with high %recovery. The method exhibits reasonable sensitivity with lower limits of detection and quantification of 5.4 and 16.6 ng mL-1, respectively, across a concentration range of 25.0-2000.0 and 50-1000 ng mL-1 for the standard mizolastine analysis and mizolastine assay in the plasma sample, respectively. Moreover, the established method was applied to assess tablet content uniformity and mizolastine assay in plasma samples with high recoveries (98.50%-100.20%). Such applications underscore the method's potential applicability within quality control laboratories, preventing the need for sample preparation or laborious extraction steps. Finally, the method's sustainability and practicality were confirmed by applying different greenness and whiteness metrics, yielding excellent results.

Asunto(s)

Espectrometría de Fluorescencia , Humanos , Azetidinas/sangre , Azetidinas/análisis , Azetidinas/química , Antagonistas de los Receptores Histamínicos/sangre , Antagonistas de los Receptores Histamínicos/análisis , Antagonistas de los Receptores Histamínicos/química , Comprimidos , Bencimidazoles/química , Bencimidazoles/sangre , Bencimidazoles/análisis , Estructura Molecular , Límite de Detección

RESUMEN

The objective of this phase 1 study was to evaluate the pharmacokinetics, safety, and tolerability of baricitinib after single and multiple doses in healthy Chinese adults. Eligible subjects received a once-daily dose of baricitinib 2, 4, or 10 mg or placebo on day 1 (single dose) and days 4 through 10 for 7 consecutive days (multiple doses). Plasma pharmacokinetic samples were collected up to 48 hours after dosing on days 1 and 10, with predose samples collected before dosing on day 1 and days 4 through 10. Safety and tolerability were also assessed. Baricitinib was rapidly absorbed, reaching peak plasma concentrations within 0.5 to 1 hour (median). Plasma concentrations declined rapidly following the attainment of peak concentrations, with a mean terminal half-life of 5.7 to 7.3 hours. Steady-state plasma concentrations of baricitinib were achieved after the second day of once-daily dosing, with minimal accumulation of baricitinib in plasma (up to 10% increase in area under the plasma concentration-time curve). Single- and multiple-dose mean values for area under the plasma concentration-time curve from time zero to infinity and maximum plasma concentration appeared to increase in an approximately dose-proportional manner across the dose range. Single and multiple oral doses of once-daily baricitinib up to 10 mg were well tolerated by healthy Chinese subjects.

Asunto(s)

Artritis Reumatoide/tratamiento farmacológico , Azetidinas/farmacocinética , Voluntarios Sanos/estadística & datos numéricos , Oxazoles/farmacocinética , Purinas/farmacocinética , Pirazoles/farmacocinética , Sulfonamidas/farmacocinética , Administración Oral , Adulto , Área Bajo la Curva , Pueblo Asiatico/etnología , Azetidinas/administración & dosificación , Azetidinas/sangre , Estudios de Casos y Controles , Relación Dosis-Respuesta a Droga , Esquema de Medicación , Tolerancia a Medicamentos , Femenino , Semivida , Humanos , Masculino , Persona de Mediana Edad , Oxazoles/administración & dosificación , Oxazoles/sangre , Placebos/administración & dosificación , Purinas/administración & dosificación , Purinas/sangre , Pirazoles/administración & dosificación , Pirazoles/sangre , Seguridad , Sulfonamidas/administración & dosificación , Sulfonamidas/sangre

RESUMEN

PF-5190457 is a selective and potent ghrelin receptor inverse agonist presently undergoing clinical trials to treat alcohol use disorder (AUD). We describe the development and validation of a selective and sensitive liquid chromatography-tandem mass spectrometry-based method for quantification of PF-5190457 and its recently discovered hydroxy metabolite PF-6870961 in human plasma. Analytes were extracted after simple protein precipitation using methanol (2.5 ng mL-1 tacrine as an internal standard). A gradient liquid chromatography method was used to separate the analytes on an Acquity UPLC BEH C18 analytical column. The separation was achieved at a flow rate of 0.25 mL min-1 and the total chromatographic runtime was 11.30 min. Positive electrospray ionization and multiple reaction monitoring mode were used for the quantification of all the analytes. The calibration curves from six validation runs were linear with a correlation coefficient of ≥0.996 for the concentration range of 1-1000 ng mL-1 and 2-250 ng mL-1 for PF-5190457 and PF-6870961, respectively. The retention time for PF-5190457, PF-6870961 and tacrine were 4.4, 3.8, and 4.6 min, respectively. The lower limit of quantification for PF-5190457 and PF-6870961 was 1 and 2 ng mL-1, respectively. The inter-assay precision and accuracy results obtained were within the Food and Drug Administration recommended ±15% limit of nominal values. All the analytes were found to be stable under varied stability conditions. The recovery of PF-5190457 and PF-6870961 ranged from 95 to 103%. Further, the application of the method was demonstrated by measuring the concentration of PF-5190457 and its hydroxy metabolite in patient plasma samples from 100 mg dose.

Asunto(s)

Azetidinas/sangre , Cromatografía Líquida de Alta Presión/métodos , Compuestos de Espiro/sangre , Espectrometría de Masas en Tándem/métodos , Adulto , Azetidinas/química , Azetidinas/metabolismo , Azetidinas/farmacocinética , Femenino , Humanos , Límite de Detección , Modelos Lineales , Masculino , Receptores de Ghrelina/agonistas , Reproducibilidad de los Resultados , Compuestos de Espiro/química , Compuestos de Espiro/metabolismo , Compuestos de Espiro/farmacocinética

RESUMEN

PURPOSE: To evaluate the PK and safety of siponimod, a substrate of CYP2C9/3A4, in the presence or absence of a CYP3A4 inhibitor, itraconazole. METHODS: This was an open-label study in healthy subjects (aged 18-50 years; genotype: CYP2C9 *1*2 [cohort 1; n = 17] or *1*3 [cohort 2; n = 13]). Subjects received siponimod 0.25-mg single dose in treatment period 1 (days 1-14), itraconazole 100 mg twice daily in treatment period 2 (days 15-18), and siponimod 0.25-mg single dose (day 19) with itraconazole until day 31 (cohort 1) or day 35 (cohort 2) in treatment period 3. PK of siponimod alone and with itraconazole and safety were assessed. RESULTS: Overall, 29/30 subjects completed the study. In treatment period 1, geometric mean AUCinf, T1/2, and median Tmax were higher while systemic clearance was lower in cohort 2 than cohort 1. In treatment period 3, siponimod AUC decreased by 10% (geo-mean ratio [90% confidence intervals]: 0.90 [0.84; 0.96]) and 24% (0.76 [0.69; 0.82]) in cohorts 1 and 2, respectively. Siponimod Cmax was similar between treatment periods 1 and 3. In both cohorts, the Cmax and AUC of the metabolites (M17, M3, and M5) decreased in the presence of itraconazole. All adverse events were mild. CONCLUSIONS: The minor albeit significant reduction in plasma exposure of siponimod and its metabolites by itraconazole was unexpected. While the reason is unclear, the results suggest that coadministration of the two drugs would not cause a considerable increase of siponimod exposure independent of CYP2C9 genotype.

Asunto(s)

Azetidinas/farmacocinética , Compuestos de Bencilo/farmacocinética , Citocromo P-450 CYP2C9/genética , Inhibidores del Citocromo P-450 CYP3A/farmacología , Citocromo P-450 CYP3A , Itraconazol/farmacología , Moduladores de los Receptores de fosfatos y esfingosina 1/farmacocinética , Adolescente , Adulto , Área Bajo la Curva , Azetidinas/efectos adversos , Azetidinas/sangre , Compuestos de Bencilo/efectos adversos , Compuestos de Bencilo/sangre , Interacciones Farmacológicas , Electrocardiografía/efectos de los fármacos , Femenino , Genotipo , Voluntarios Sanos , Humanos , Recuento de Linfocitos , Masculino , Persona de Mediana Edad , Moduladores de los Receptores de fosfatos y esfingosina 1/efectos adversos , Moduladores de los Receptores de fosfatos y esfingosina 1/sangre , Adulto Joven

RESUMEN

A sensitive and selective method of high performance liquid chromatography (HPLC) coupled to tandem mass spectrometry (MS/MS) has been developed for the simultaneous quantification of six anticancer protein kinase inhibitors (PKIs), dabrafenib, trametinib, vemurafenib, cobimetinib, pazopanib, regorafenib, and two active metabolites (regorafenib-M2 and regorafenib-M5) in human plasma. Plasma protein precipitation with methanol enables the sample extraction of 100 µL aliquot of plasma. Analytes are detected by electrospray triple-stage quadrupole mass spectrometry and quantified using the calibration curves with stable isotope-labeled internal standards. The method was validated based on FDA recommendations, including assessment of extraction yield (74-104%), matrix effects, analytical recovery (94-104%) with low variability (<15%). The method is sensitive (lower limits of quantification within 1 to 200 ng/mL), accurate (intra- and inter-assay bias: -0.3% to +12.7%, and -3.2% to +6.3%, respectively) and precise (intra- and inter-assay CVs within 0.7-7.3% and 2.5-8.0%, respectively) over the clinically relevant concentration range (upper limits of quantification 500 to 100,000 ng/mL). This method is applied in our laboratory for both clinical research programs and routine therapeutic drug monitoring service of PKIs.

Asunto(s)

Antineoplásicos/sangre , Cromatografía Líquida de Alta Presión/métodos , Espectrometría de Masas en Tándem/métodos , Administración Oral , Antineoplásicos/administración & dosificación , Antineoplásicos/química , Antineoplásicos/farmacocinética , Azetidinas/administración & dosificación , Azetidinas/sangre , Azetidinas/química , Azetidinas/farmacocinética , Niño , Humanos , Imidazoles/administración & dosificación , Imidazoles/sangre , Imidazoles/química , Imidazoles/farmacocinética , Indazoles , Indoles/administración & dosificación , Indoles/sangre , Indoles/química , Indoles/farmacocinética , Límite de Detección , Modelos Lineales , Oximas/administración & dosificación , Oximas/sangre , Oximas/química , Oximas/farmacocinética , Compuestos de Fenilurea/administración & dosificación , Compuestos de Fenilurea/sangre , Compuestos de Fenilurea/química , Compuestos de Fenilurea/farmacocinética , Piperidinas/administración & dosificación , Piperidinas/sangre , Piperidinas/química , Piperidinas/farmacocinética , Piridinas/administración & dosificación , Piridinas/sangre , Piridinas/química , Piridinas/farmacocinética , Piridonas/administración & dosificación , Piridonas/sangre , Piridonas/química , Piridonas/farmacocinética , Pirimidinas/administración & dosificación , Pirimidinas/sangre , Pirimidinas/química , Pirimidinas/farmacocinética , Pirimidinonas/administración & dosificación , Pirimidinonas/sangre , Pirimidinonas/química , Pirimidinonas/farmacocinética , Reproducibilidad de los Resultados

RESUMEN

Baricitinib, an oral selective Janus kinase 1 and 2 inhibitor, undergoes active renal tubular secretion. Baricitinib was not predicted to inhibit hepatic and renal uptake and efflux drug transporters, based on the ratio of the unbound maximum eliminating-organ inlet concentration and the in vitro half-maximal inhibitory concentrations (IC50 ). In vitro, baricitinib was a substrate for organic anion transporter (OAT)3, multidrug and toxin extrusion protein (MATE)2-K, P-glycoprotein (P-gp), and breast cancer resistance protein (BCRP). Probenecid, a strong OAT3 inhibitor, increased the area under the concentration-time curve from time zero to infinity (AUC[0-∞] ) of baricitinib by twofold and decreased renal clearance to 69% of control in healthy subjects. Physiologically based pharmacokinetic (PBPK) modeling reproduced the renal clearance of baricitinib and the inhibitory effect of probenecid using the in vitro IC50 value of 4.4 µM. Using ibuprofen and diclofenac in vitro IC50 values of 4.4 and 3.8 µM toward OAT3, 1.2 and 1.0 AUC(0-∞) ratios of baricitinib were predicted. These predictions suggest clinically relevant drug-drug interactions (DDIs) with ibuprofen and diclofenac are unlikely.

Asunto(s)

Azetidinas/farmacología , Proteínas de Transporte de Membrana/metabolismo , Sulfonamidas/farmacología , Adulto , Área Bajo la Curva , Azetidinas/sangre , Azetidinas/farmacocinética , Interacciones Farmacológicas , Células HEK293 , Humanos , Masculino , Persona de Mediana Edad , Purinas , Pirazoles , Sulfonamidas/sangre , Sulfonamidas/farmacocinética , Factores de Tiempo , Adulto Joven

RESUMEN

Targeted therapies for cancers are fast-growing therapies. For instance kinase inhibitors such as BRAF inhibitors (BRAFi) and MEK inhibitors (MEKi) are increasingly used to treat malignant melanoma. The metabolic profile of these drugs can result in great interindividual variability, justifying therapeutic drug monitoring (TDM). We describe a rapid and specific method for quantification of 2 BRAFi (vemurafenib, dabrafenib) and 3 MEKi (cobimetinib, trametinib and binimetinib). Chromatography was performed on a Waters Acquity-UPLC system with CORTECS C18+ column, under a gradient of 10% acetic acid in water/acetonitrile. An Acquity-TQD® with electrospray ionization was used for detection. Samples were prepared by solid phase extraction (Oasis® MCX microElution) before injection in the system. Calibration curves ranges from 0.4 to 100µg/ml for vemurafenib, from 1 to 1000ng/ml for dabrafenib, from 0.5 to 500ng/ml for cobimetinib and binimetinib, and from 0.75 to 250ng/ml for trametinib. At all concentrations the bias was within ±15% of the nominal concentrations and precision was ≤15%. All results were within the acceptance criteria of the EMA guidelines on method validation. This rapid, sensitive and specific UPLC-MS/MS method can perform simultaneous quantification of targeted therapies used in malignant melanoma and is usable for routine TDM.

Asunto(s)

Cromatografía Líquida de Alta Presión/métodos , Melanoma/sangre , Melanoma/tratamiento farmacológico , Quinasas de Proteína Quinasa Activadas por Mitógenos/antagonistas & inhibidores , Inhibidores de Proteínas Quinasas/sangre , Proteínas Proto-Oncogénicas B-raf/antagonistas & inhibidores , Espectrometría de Masas en Tándem/métodos , Azetidinas/sangre , Bencimidazoles/sangre , Humanos , Imidazoles/sangre , Imidazoles/uso terapéutico , Indoles/sangre , Indoles/uso terapéutico , Límite de Detección , Modelos Lineales , Melanoma/enzimología , Oximas/sangre , Oximas/uso terapéutico , Piperidinas/sangre , Inhibidores de Proteínas Quinasas/uso terapéutico , Piridonas/sangre , Piridonas/uso terapéutico , Pirimidinonas/sangre , Pirimidinonas/uso terapéutico , Sulfonamidas/sangre , Sulfonamidas/uso terapéutico , Vemurafenib

RESUMEN

OBJECTIVE: To assess the pharmacokinetics (PK), safety, and tolerability of siponimod and major metabolites in subjects with mild, moderate, and severe hepatic impairment (HI) compared with demographically-matched healthy subjects (HS). METHODS: This open-label, parallel-group study enrolled 40 subjects (each HI group, n = 8; HS group, n = 16). A staged design was employed starting with the enrollment of subjects with mild HI, followed by those with moderate and severe HI. All subjects received single oral doses of 0.25 mg siponimod on day 1; PK and safety data were collected during the 21-day follow-up. RESULTS: All subjects had similar baseline characteristics and completed the study. No significant differences were observed in the plasma exposure of siponimod in mild, moderate, and severe HI groups vs. HS: Cmax changed by 16%, -13%, and -16%; AUC by 5%, -13%, and 15%, respectively. The unbound siponimod PK parameters vs. HS were similar in the mild HI, and increased in the moderate (Cmax, 15%; AUC, 17%) and severe HI groups (Cmax, 11%; AUC, 50%). Exposure of M3 and M5 also showed 2- to 5-fold increase, particularly in the moderate and severe HI groups vs HS. There were no clinically-relevant safety findings. CONCLUSIONS: Single oral doses of 0.25 mg siponimod were well tolerated, and HI did not significantly alter exposure to siponimod. Increase in the M3 and M5 metabolites requires further evaluation. These results do not warrant any dose adjustments of siponimod in subjects with HI.
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Asunto(s)

Azetidinas/efectos adversos , Azetidinas/farmacocinética , Compuestos de Bencilo/efectos adversos , Compuestos de Bencilo/farmacocinética , Insuficiencia Hepática/metabolismo , Hígado/efectos de los fármacos , Administración Oral , Adolescente , Adulto , Anciano , Área Bajo la Curva , Azetidinas/sangre , Azetidinas/metabolismo , Compuestos de Bencilo/sangre , Compuestos de Bencilo/metabolismo , Femenino , Semivida , Insuficiencia Hepática/sangre , Insuficiencia Hepática/diagnóstico , Humanos , Hígado/metabolismo , Masculino , Persona de Mediana Edad , Receptores de Lisoesfingolípidos/metabolismo , Índice de Severidad de la Enfermedad , Adulto Joven

RESUMEN

OBJECTIVE: To investigate the pharmacokinetics (PK), safety, and tolerability of siponimod and selected inactive metabolites (M3 and M5) in subjects with varying degrees of renal impairment (RI) compared to demographically matched healthy subjects (HS). METHODS: The study enrolled subjects with severe RI (n = 8) and matched HS (n = 8). Subjects with moderate and mild RI were to be enrolled only if interim analysis showed ≥ 50% increase in maximum plasma concentration (Cmax) or area under the curve (AUC) of total and/or unbound siponimod in severe RI subjects vs. HS. All subjects received a single oral dose of siponimod 0.25 mg on day 1; PK and safety were evaluated during the follow-up (~ 13 days). RESULTS: PK of siponimod was marginally affected in severe RI subjects vs. HS: Cmax decreased by 8%, and AUClast and AUCinf increased by 23% and 24%, respectively; half-life (37 vs. 26 hours) and systemic clearance (2.9 vs. 3.4 L/h) were comparable. Siponimod plasma unbound (u) fraction at 4 hours post-dose was similar between the two groups (range: 0.0172 - 0.0550%). Cmax(u) was comparable while AUClast(u) and AUCinf(u) were increased by 33% compared to HS. M3 exposure was similar (Cmax decreased by 9%; AUClast and AUCinf increased by 11%) and M5 exposure was slightly lower (Cmax decreased by 26%; AUClast decreased by 16%) in subjects with severe renal impairment (RI) compared with matched HS. No adverse events were reported during this study. CONCLUSIONS: Changes in the plasma exposure of total and unbound siponimod and metabolites M3 and M5 were not considered to be clinically relevant. Further to severe RI, investigation of PK in subjects with mild and moderate RI was not warranted.
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Asunto(s)

Azetidinas/efectos adversos , Azetidinas/farmacocinética , Compuestos de Bencilo/efectos adversos , Compuestos de Bencilo/farmacocinética , Insuficiencia Renal/metabolismo , Administración Oral , Adolescente , Adulto , Anciano , Área Bajo la Curva , Azetidinas/sangre , Azetidinas/metabolismo , Compuestos de Bencilo/sangre , Compuestos de Bencilo/metabolismo , Femenino , Semivida , Humanos , Masculino , Persona de Mediana Edad , Receptores de Lisoesfingolípidos/metabolismo , Insuficiencia Renal/sangre , Insuficiencia Renal/diagnóstico , Índice de Severidad de la Enfermedad , Adulto Joven

RESUMEN

UNLABELLED: This was a first-in-human study of the novel phosphodiesterase-2A (PDE2A) PET ligand (18)F-PF-05270430. The primary goals were to determine the appropriate tracer kinetic model to quantify brain uptake and to examine the within-subject test-retest variability. METHODS: In advance of human studies, radiation dosimetry was determined in nonhuman primates. Six healthy male subjects participated in a test-retest protocol with dynamic scans and metabolite-corrected input functions. Nine brain regions of interest were studied, including the striatum, white matter, neocortical regions, and cerebellum. Multiple modeling methods were applied to calculate volume of distribution (VT) and binding potentials relative to the nondisplaceable tracer in tissue (BPND), concentration of tracer in plasma (BPP), and free tracer in tissue (BPF). The cerebellum was selected as a reference region to calculate binding potentials. RESULTS: The dosimetry study provided an effective dose of less than 0.30 mSv/MBq, with the gallbladder as the critical organ; the human target dose was 185 MBq. There were no adverse events or clinically detectable pharmacologic effects reported. Tracer uptake was highest in the striatum, followed by neocortical regions and white matter, and lowest in the cerebellum. Regional time-activity curves were well fit by multilinear analysis-1, and a 70-min scan duration was sufficient to quantify VT and the binding potentials. BPND, with mean values ranging from 0.3 to 0.8, showed the best intrasubject and intersubject variability and reliability. Test-retest variability in the whole brain (excluding the cerebellum) of VT, BPND, and BPP were 8%, 16%, and 17%, respectively. CONCLUSION: (18)F-PF-05270430 shows promise as a PDE2A PET ligand, albeit with low binding potential values.

Asunto(s)

Compuestos de Azabiciclo/farmacocinética , Azetidinas/farmacocinética , Encéfalo/diagnóstico por imagen , Encéfalo/metabolismo , Fosfodiesterasas de Nucleótidos Cíclicos Tipo 2/metabolismo , Modelos Biológicos , Tomografía de Emisión de Positrones/métodos , Animales , Compuestos de Azabiciclo/sangre , Azetidinas/sangre , Simulación por Computador , Estudios de Factibilidad , Femenino , Humanos , Marcaje Isotópico , Macaca mulatta , Masculino , Tasa de Depuración Metabólica , Imagen Molecular/métodos , Especificidad de Órganos , Proyectos Piloto , Radiofármacos/sangre , Radiofármacos/farmacocinética , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Distribución Tisular

RESUMEN

The pharmacokinetics, metabolism, and excretion of cobimetinib, a MEK inhibitor, were characterized in healthy male subjects (n = 6) following a single 20 mg (200 µCi) oral dose. Unchanged cobimetinib and M16 (glycine conjugate of hydrolyzed cobimetinib) were the major circulating species, accounting for 20.5% and 18.3% of the drug-related material in plasma up to 48 hours postdose, respectively. Other circulating metabolites were minor, accounting for less than 10% of drug-related material in plasma. The total recovery of the administered radioactivity was 94.3% (±1.6%, S.D.) with 76.5% (±2.3%) in feces and 17.8% (±2.5%) in urine. Metabolite profiling indicated that cobimetinib had been extensively metabolized with only 1.6% and 6.6% of the dose remaining as unchanged drug in urine and feces, respectively. In vitro phenotyping experiments indicated that CYP3A4 was predominantly responsible for metabolizing cobimetinib. From this study, we concluded that cobimetinib had been well absorbed (fraction absorbed, Fa = 0.88). Given this good absorption and the previously determined low hepatic clearance, the systemic exposures were lower than expected (bioavailability, F = 0.28). We hypothesized that intestinal metabolism had strongly attenuated the oral bioavailability of cobimetinib. Supporting this hypothesis, the fraction escaping gut wall elimination (Fg) was estimated to be 0.37 based on F and Fa from this study and the fraction escaping hepatic elimination (Fh) from the absolute bioavailability study (F = Fa × Fh × Fg). Physiologically based pharmacokinetics modeling also showed that intestinal clearance had to be included to adequately describe the oral profile. These collective data suggested that cobimetinib was well absorbed following oral administration and extensively metabolized with intestinal first-pass metabolism contributing to its disposition.

Asunto(s)

Antineoplásicos/administración & dosificación , Antineoplásicos/farmacocinética , Azetidinas/administración & dosificación , Azetidinas/farmacocinética , Absorción Intestinal , Mucosa Intestinal/metabolismo , Quinasas de Proteína Quinasa Activadas por Mitógenos/antagonistas & inhibidores , Piperidinas/administración & dosificación , Piperidinas/farmacocinética , Inhibidores de Proteínas Quinasas/administración & dosificación , Inhibidores de Proteínas Quinasas/farmacocinética , Eliminación Renal , Administración Oral , Adulto , Antineoplásicos/sangre , Antineoplásicos/orina , Azetidinas/sangre , Azetidinas/orina , Disponibilidad Biológica , Biotransformación , Radioisótopos de Carbono , Citocromo P-450 CYP3A/metabolismo , Heces/química , Glicina/metabolismo , Voluntarios Sanos , Humanos , Hidrólisis , Intestinos/enzimología , Masculino , Microsomas Hepáticos/metabolismo , Persona de Mediana Edad , Quinasas de Proteína Quinasa Activadas por Mitógenos/metabolismo , Modelos Biológicos , Piperidinas/sangre , Piperidinas/orina , Inhibidores de Proteínas Quinasas/sangre , Inhibidores de Proteínas Quinasas/orina , Especificidad por Sustrato , Adulto Joven

RESUMEN

PURPOSE: To characterize cobimetinib pharmacokinetics and evaluate impact of clinically relevant covariates on cobimetinib pharmacokinetics. METHODS: Plasma samples (N = 4886) were collected from 487 patients with various solid tumors (mainly melanoma) in three clinical studies (MEK4592g, NO25395, GO28141). Cobimetinib was administered orally, once daily on either a 21-day-on/7-day-off, 14-day-on/14-day-off or 28-day-on schedule in a 28-day dosing cycle as single agent or in combination with vemurafenib. Cobimetinib doses ranged from 2.1 to 125 mg. NONMEM was used for pharmacokinetic analysis. RESULTS: A linear two-compartment model with first-order absorption, lag time and first-order elimination described cobimetinib pharmacokinetics. The typical estimates (inter-individual variability) of apparent clearance (CL/F), central volume of distribution (V2/F) and terminal half-life were 322 L/day (58 %), 511 L (49 %) and 2.2 days, respectively. Inter-occasion variability on relative bioavailability was estimated at 46 %. CL/F decreased with age. V2/F increased with body weight (BWT). However, the impact of age and BWT on cobimetinib steady-state exposure (peak and trough concentrations and AUC following the recommended daily dose of 60 mg 21-day-on/7-day-off) was limited (<25 % changes across the distribution of age and BWT). No significant difference in cobimetinib pharmacokinetics or steady-state exposure was observed between patient subgroups based on sex, renal function, ECOG score, hepatic function tests, race, region, cancer type, and co-administration of moderate and weak CYP3A inducers or inhibitors and vemurafenib. CONCLUSION: A population pharmacokinetic model was developed for cobimetinib in cancer patients. Covariates had minimal impact on steady-state exposure, suggesting no need for dose adjustments and supporting the recommended dose for all patients.

Asunto(s)

Antineoplásicos/farmacocinética , Azetidinas/farmacocinética , Neoplasias/tratamiento farmacológico , Piperidinas/farmacocinética , Inhibidores de Proteínas Quinasas/farmacocinética , Adulto , Anciano , Anciano de 80 o más Años , Antineoplásicos/sangre , Antineoplásicos/uso terapéutico , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Azetidinas/sangre , Azetidinas/uso terapéutico , Peso Corporal , Citocromo P-450 CYP3A/metabolismo , Esquema de Medicación , Femenino , Glucuronosiltransferasa/metabolismo , Semivida , Humanos , Inactivación Metabólica/efectos de los fármacos , Indoles/administración & dosificación , Riñón/fisiopatología , Hígado/fisiopatología , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Masculino , Persona de Mediana Edad , Modelos Biológicos , Proteínas de Neoplasias/antagonistas & inhibidores , Neoplasias/sangre , Piperidinas/sangre , Piperidinas/uso terapéutico , Inhibidores de Proteínas Quinasas/sangre , Inhibidores de Proteínas Quinasas/uso terapéutico , Sulfonamidas/administración & dosificación , Vemurafenib , Adulto Joven

RESUMEN

PF-5190457 is a ghrelin receptor inverse agonist that is currently undergoing clinical development for the treatment of alcoholism. Our aim was to develop and validate a simple and sensitive assay for quantitative analysis of PF-5190457 in human or rat plasma and rat brain using liquid chromatography-tandem mass spectrometry. The analyte and stable isotope internal standard were extracted from 50 µL plasma or rat brain homogenate by protein precipitation using 0.1% formic acid in acetonitrile. Chromatography was carried out on an Acquity UPLC BEH C18 (2.1 mm × 50 mm) column with 1.7 µm particle size and 130 Å pore size. The flow rate was 0.5 mL/min and total chromatographic run time was 2.2 min. The mobile phase consisted of a gradient mixture of water: acetonitrile 95:5% (v/v) containing 0.1% formic acid (solvent A) and 100% acetonitrile containing 0.1% formic acid (solvent B). Multiple reaction monitoring was carried out in positive electro-spray ionization mode using m/z 513.35 → 209.30 for PF-5190457 and m/z 518.47 → 214.43 for the internal standard. The recovery ranged from 102 to 118% with coefficient of variation (CV) less than 6% for all matrices. The calibration curves for all matrices were linear over the studied concentration range (R(2) ≥ 0.998, n = 3). The lower limit of quantification was 1 ng/mL in rat or human plasma and 0.75 ng/g in rat brain. Intra- and inter-run mean percent accuracies were between 85 and 115% and percent imprecision was ≤15%. The assays were successfully utilized to measure the concentration of PF-5190457 in pre-clinical and clinical pharmacology studies of the compound.

Asunto(s)

Encéfalo/metabolismo , Cromatografía Liquida/métodos , Receptores de Ghrelina/antagonistas & inhibidores , Espectrometría de Masas en Tándem/métodos , Animales , Azetidinas/sangre , Azetidinas/metabolismo , Humanos , Límite de Detección , Ratas , Compuestos de Espiro/sangre , Compuestos de Espiro/metabolismo

RESUMEN

This randomized, double-blind, placebo-controlled, 6-arm, parallel-design study investigated cardiac and hematological pharmacodynamic effects of ceralifimod (ONO-4641), a selective sphingosine-1-phosphate (S1P) receptor modulator, over a broad dose range in direct comparison with the nonselective S1P modulator fingolimod. Healthy subjects were assigned to ceralifimod (0.01, 0.025, 0.05, or 0.10 mg), fingolimod (0.5 mg), or placebo once daily for 14 days (n = 24 per group). After 14 days of treatment, mean absolute lymphocyte count percentage change from baseline was greatest in the fingolimod (-62%) and ceralifimod 0.10 mg (-56%) groups. On treatment cessation, lymphocyte recovery was faster in the ceralifimod versus the fingolimod group. Ceralifimod showed dose- and concentration-dependent chronotropic effect. Cardiac effects in the fingolimod group were dependent on fingolimod-P concentrations. Maximum mean heart rate (HR) effect on day 1 was larger with fingolimod (placebo-adjusted change from time-matched baseline HR [ΔΔHR], -14.9 beats per minute [bpm]) versus ceralifimod (ΔΔHR, -6.2 and -12.0 bpm for the 0.05- and 0.10-mg doses, respectively). Ceralifimod's effect on the PR interval was minor. Safety biomarker results suggest that potential therapeutic doses of ceralifimod, in particular the 0.05-mg dose, might result in reduced occurrence of bradycardia, atrioventricular block absolute lymphocyte count and grade 3/4 lymphopenia compared with fingolimod 0.5 mg.

Asunto(s)

Azetidinas/farmacocinética , Clorhidrato de Fingolimod/farmacocinética , Corazón/efectos de los fármacos , Inmunosupresores/farmacocinética , Linfocitos/efectos de los fármacos , Naftalenos/farmacocinética , Adolescente , Adulto , Área Bajo la Curva , Azetidinas/administración & dosificación , Azetidinas/sangre , Relación Dosis-Respuesta a Droga , Método Doble Ciego , Femenino , Clorhidrato de Fingolimod/administración & dosificación , Clorhidrato de Fingolimod/sangre , Semivida , Humanos , Inmunosupresores/administración & dosificación , Inmunosupresores/sangre , Masculino , Persona de Mediana Edad , Naftalenos/administración & dosificación , Naftalenos/sangre , Adulto Joven

RESUMEN

A liquid chromatography-tandem mass spectrometry method (LC-MS/MS) was developed to quantify ezetimibe (EZM) and its major glucuronide (ezetimibe glucuronide, EZM-G) in human plasma simultaneously. The analytes were purified by solid phase extraction (SPE) without hydrolysis. Separation of the analytes was achieved using acetonitrile-water (0.08% formic acid) (70:30, v/v) as the mobile phase at a flow rate of 0.8 mL/min on an Agilent Extend C18 column. The analytes were detected by LC-MS/MS using negative ionization in multiple reaction monitoring (MRM) mode. The mass transition pairs of m/z 408.4→271.0 and m/z 584.5→271.0 were used to detect EZM and EZM-G, respectively. The analytical method was linear over the concentration range of 0.1-20 ng/mL for EZM and 0.5-200 ng/mL for EZM-G. Within- and between-run precision for EZM was no more than 8.6% and 12.8%; and for EZM-G was no more than 9.0% and 8.7%, respectively. This method was reproducible and reliable, and was successfully used to analyze human plasma samples for application in a bioequivalence study.

Asunto(s)

Azetidinas/sangre , Azetidinas/farmacocinética , Cromatografía Liquida/métodos , Ezetimiba/sangre , Ezetimiba/farmacocinética , Glucurónidos/sangre , Glucurónidos/farmacocinética , Espectrometría de Masas en Tándem/métodos , Azetidinas/química , Ezetimiba/química , Glucurónidos/química , Humanos , Modelos Lineales , Masculino , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Extracción en Fase Sólida

RESUMEN

Data from the clinical absolute bioavailability (F) study with cobimetinib suggested that F was lower than predicted based on its low hepatic extraction and good absorption. The CYP3A4 transgenic (Tg) mouse model with differential expression of CYP3A4 in the liver (Cyp3a(-/-)Tg-3A4Hep) or intestine (Cyp3a(-/-)Tg-3A4Int) and both liver and intestine (Cyp3a(-/-)Tg-3A4Hep/Int) were used to study the contribution of intestinal metabolism to the F of cobimetinib. In addition, the effect of CYP3A4 inhibition and induction on cobimetinib exposures was tested in the Cyp3a(-/-)Tg-3A4Hep/Int and PXR-CAR-CYP3A4/CYP3A7 mouse models, respectively. After i.v. administration of 1 mg/kg cobimetinib to wild-type [(WT) FVB], Cyp3a(-/-)Tg-3A4Hep, Cyp3a(-/-)Tg-3A4Int, or Cyp3a(-/-)Tg-3A4Hep/Int mice, clearance (CL) (26-35 ml/min/kg) was similar in the CYP3A4 transgenic and WT mice. After oral administration of 5 mg/kg cobimetinib, the area under the curve (AUC) values of cobimetinib in WT, Cyp3a(-/-)Tg-3A4Hep, Cyp3a(-/-)Tg-3A4Int, or Cyp3a(-/-)Tg-3A4Hep/Int mice were 1.35, 3.39, 1.04, and 0.701 µM⋅h, respectively. The approximately 80% lower AUC of cobimetinib in transgenic mice when intestinal CYP3A4 was present suggested that the intestinal first pass contributed to the oral CL of cobimetinib. Oxidative metabolites observed in human circulation were also observed in the transgenic mice. In drug-drug interaction (DDI) studies using Cyp3a(-/-)Tg-3A4Hep/Int mice, 8- and 4-fold increases in oral and i.v. cobimetinib exposure, respectively, were observed with itraconazole co-administration. In PXR-CAR-CYP3A4/CYP3A7 mice, rifampin induction decreased cobimetinib oral exposure by approximately 80%. Collectively, these data support the conclusion that CYP3A4 intestinal metabolism contributes to the oral disposition of cobimetinib and suggest that under certain circumstances the transgenic model may be useful in predicting clinical DDIs.

Asunto(s)

Antineoplásicos/farmacocinética , Azetidinas/farmacocinética , Citocromo P-450 CYP3A/metabolismo , Mucosa Intestinal/enzimología , MAP Quinasa Quinasa 1/antagonistas & inhibidores , Microsomas Hepáticos/enzimología , Piperidinas/farmacocinética , Inhibidores de Proteínas Quinasas/farmacocinética , Administración Oral , Animales , Antineoplásicos/administración & dosificación , Antineoplásicos/sangre , Hidrocarburo de Aril Hidroxilasas/genética , Hidrocarburo de Aril Hidroxilasas/metabolismo , Azetidinas/administración & dosificación , Azetidinas/sangre , Disponibilidad Biológica , Citocromo P-450 CYP3A/química , Citocromo P-450 CYP3A/genética , Inductores del Citocromo P-450 CYP3A/efectos adversos , Inhibidores del Citocromo P-450 CYP3A/efectos adversos , Evaluación Preclínica de Medicamentos , Interacciones Farmacológicas , Femenino , Semivida , Humanos , Inyecciones Intravenosas , Mucosa Intestinal/efectos de los fármacos , Mucosa Intestinal/metabolismo , MAP Quinasa Quinasa 1/metabolismo , Tasa de Depuración Metabólica , Ratones Noqueados , Ratones Transgénicos , Microsomas Hepáticos/efectos de los fármacos , Microsomas Hepáticos/metabolismo , Piperidinas/administración & dosificación , Piperidinas/sangre , Inhibidores de Proteínas Quinasas/administración & dosificación , Inhibidores de Proteínas Quinasas/sangre , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo

RESUMEN

Inhibition of MAP/ERK kinase (MEK) is a promising strategy to control the growth of tumors that are dependent on aberrant signaling in the MEK pathway. Cobimetinib (GDC-0973) (S)-[3,4-Difluoro-2-(2-fluoro-4-iodo-phenylamino)-phenyl]-((S)-3-hydroxy-3-piperidin-2-yl-azetidin-1-yl)-methanone) inhibits proliferation of a variety of human tumor cell lines by inhibiting MEK1 and MEK2. A specific high performance liquid chromatography-mass spectrometric assay was developed and validated for the determination of cobimetinib in human plasma. The overall mean recovery using protein precipitation extraction with acetonitrile was found to be 54.1%. The calibration curve was ranged from 0.20 to 100ng/mL. The LLOQ was sensitive enough to detect terminal phase concentrations of the drug. The intra- and inter-assay precision (%CV) was within 10.3% and 9.5% for cobimetinib. The assay accuracy (%RE) was within ±13.7% of the nominal concentration values for cobimetinib with the normal analytical QCs. The developed assay was successfully used to analyze the human plasma samples (for pharmacokinetic analysis) from clinical trials.

Asunto(s)

Azetidinas/sangre , Cromatografía Líquida de Alta Presión/métodos , MAP Quinasa Quinasa 1/antagonistas & inhibidores , Espectrometría de Masas/métodos , Piperidinas/sangre , Inhibidores de Proteínas Quinasas/sangre , Proteínas/química , Azetidinas/farmacocinética , Línea Celular Tumoral , Estudios Cruzados , Humanos , Límite de Detección , Piperidinas/farmacocinética , Inhibidores de Proteínas Quinasas/farmacocinética , Reproducibilidad de los Resultados

RESUMEN

The anticancer activity of a novel pure 1,4-Diaryl-2-azetidinone (1), endowed with a higher solubility than the well known Combretastatin A4, is tested in mice. We previously reported that Compound (1) showed specific antiproliferative activity against duodenal and colon cancer cells, inducing activation of AMP-activated protein kinase and apoptosis. Here we estimate that the maximum tolerated dose in a mouse model is 40 mg/kg; the drug is well tolerated both in single dose and in repeated administration schedules. The drug displays a significant antitumor activity and a tumor growth delay when administered at the MTD both in single and fractionated i.v. administration in a mouse xenograft model of colorectal cancer. Arrest of tumor growth and relapse after drug suspension are parallel to modification in glucose demand as shown by PET studies with [(18)F] FDG. These data strongly support Compound (1) as a promising molecule for in vivo treatment of colorectal cancer.

Asunto(s)

Proteínas Quinasas Activadas por AMP/metabolismo , Antineoplásicos , Azetidinas , Neoplasias Colorrectales/tratamiento farmacológico , Glucosa/metabolismo , Guayacol/análogos & derivados , Animales , Antineoplásicos/sangre , Antineoplásicos/farmacocinética , Antineoplásicos/farmacología , Antineoplásicos/uso terapéutico , Azetidinas/sangre , Azetidinas/farmacocinética , Azetidinas/farmacología , Azetidinas/uso terapéutico , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Neoplasias Colorrectales/metabolismo , Neoplasias Colorrectales/patología , Guayacol/sangre , Guayacol/farmacocinética , Guayacol/farmacología , Guayacol/uso terapéutico , Humanos , Masculino , Ratones , Ensayos Antitumor por Modelo de Xenoinjerto

RESUMEN

Baricitinib (also known as LY3009104 or INCB028050), a novel and potent small molecule inhibitor of Janus kinase family of enzymes (JAKs) with selectivity for JAK1 and JAK2, is currently in clinical development for the treatment of rheumatoid arthritis (RA) and other inflammatory disorders. Two double-blind, randomized, and placebo-controlled studies were conducted to evaluate single ascending doses of 1-20 mg and multiple ascending doses of 2-20 mg QD and 5 mg BID for 10 or 28 days in healthy volunteers. Following oral administration, baricitinib plasma concentration typically attains its peak value within 1.5 hours postdose and subsequently declines in a bi-exponential fashion. Baricitinib demonstrates dose-linear and time-invariant pharmacokinetics, with low oral-dose clearance (17 L/h) and minimal systemic accumulation observed following repeat dosing. The mean renal clearance of baricitinib was determined to be ∼2 L/h. The effect of a high-fat meal on baricitinib pharmacokinetics was insignificant. The pharmacodynamics of baricitinib, evaluated by the inhibition of STAT3 phosphorylation following cytokine stimulation in the whole blood ex vivo, was well correlated with baricitinib plasma concentrations. Baricitinib was generally safe and well tolerated, with no serious treatment-related adverse events (AEs) reported from either of the studies. An expected rapidly reversible, dose-related decline in absolute neutrophil count was seen with baricitinib.

Asunto(s)

Azetidinas , Inhibidores de Proteínas Quinasas , Sulfonamidas , Adolescente , Adulto , Azetidinas/efectos adversos , Azetidinas/sangre , Azetidinas/farmacocinética , Azetidinas/farmacología , Relación Dosis-Respuesta a Droga , Método Doble Ciego , Femenino , Interacciones Alimento-Droga , Voluntarios Sanos , Humanos , Janus Quinasa 1/antagonistas & inhibidores , Janus Quinasa 2/antagonistas & inhibidores , Masculino , Persona de Mediana Edad , Inhibidores de Proteínas Quinasas/efectos adversos , Inhibidores de Proteínas Quinasas/sangre , Inhibidores de Proteínas Quinasas/farmacocinética , Inhibidores de Proteínas Quinasas/farmacología , Purinas , Pirazoles , Factor de Transcripción STAT3/antagonistas & inhibidores , Sulfonamidas/efectos adversos , Sulfonamidas/sangre , Sulfonamidas/farmacocinética , Sulfonamidas/farmacología , Adulto Joven

RESUMEN

Glucokinase activators (GKAs) are being developed for the treatment of type 2 diabetes. The toxicity of 4 GKAs (PF-04279405, PF-04651887, piragliatin, and PF-04937319) was assessed in mice, rats, dogs, and/or monkeys. GKAs were administered for 2 to 8 weeks. Standard endpoints, glucose, and insulin were assessed. All compounds produced varying degrees of hypoglycemia in all species. Brain neuronal necrosis and/or peripheral neuropathy were observed with most compounds. These findings are consistent with literature reports linking hypoglycemia with nervous system effects. Arteriopathy, mainly of cardiac vessels, was observed at a low frequency in monkey and/or dog. Arteriopathy occurred only at doses that produced severe and prolonged periods of repeated hypoglycemia. Since this lesion occurred in multiple studies with structurally distinct GKAs, these results suggested arteriopathy was related to GKA pharmacology. The morphological characteristics of the arteriopathy were consistent with that produced by experimental catecholamine administration. We hypothesize that the prolonged periods of hypoglycemia resulted in increased local and/or systemic concentrations of catecholamines via a counterregulatory and/or stress-related mechanism. Alternatively, prolonged hypoglycemia may have resulted in endothelial dysfunction leading to arteriopathy. This risk can be managed in human patients in clinical studies by careful glucose monitoring and intervention to avoid prolonged episodes of hypoglycemia.

Asunto(s)

Azetidinas/efectos adversos , Bencenoacetamidas/efectos adversos , Benzofuranos/efectos adversos , Hipoglucemia/patología , Necrosis/patología , Enfermedades del Sistema Nervioso Periférico/patología , Pirimidinas/efectos adversos , Animales , Azetidinas/sangre , Bencenoacetamidas/sangre , Benzofuranos/sangre , Cromatografía Líquida de Alta Presión , Perros , Evaluación Preclínica de Medicamentos , Femenino , Hipoglucemia/inducido químicamente , Hipoglucemiantes/efectos adversos , Insulina/sangre , Macaca fascicularis , Masculino , Ratones , Ratones Endogámicos ICR , Necrosis/inducido químicamente , Neuronas/efectos de los fármacos , Neuronas/patología , Enfermedades del Sistema Nervioso Periférico/inducido químicamente , Pirimidinas/sangre , Ratas , Ratas Sprague-Dawley