RESUMEN
A method coupling hydrophilic interaction chromatography (HILIC) with tandem mass spectrometry (MS/MS) has been developed for the simultaneous determination of three polar non-structurally related compounds--a carbapenem antibiotic, imipenem (IMP), a renal dehydropeptidase inhibitor, cilastatin (CIL), and an investigational beta-lactamase inhibitor, MK-4698 (BLI), in rat plasma, monkey plasma and mouse blood. The analytes were extracted through protein precipitation, chromatographed on a Waters Atlantis HILIC column, and detected on a Sciex API4000 mass spectrometer using a Turbo-Ion Spray ion source in positive ionization mode following multiple-reaction monitoring (MRM). The assay dynamic range was 0.1-100 microg/mL for IMP, CIL and BLI, respectively, using a total of 20-25 microL biologic samples, and the total HPLC/MS/MS run time was 4 min/injection. The assay was found to be sensitive, selective and reproducible. The challenges, namely, sample stability, blood sample processing, matrix effect in monkey study samples, and dilution re-assays for the limited mouse blood samples, are resolved and discussed. This technique allowed rapid analysis of polar compounds in biologic matrixes with satisfactory chromatographic retention and increased throughput.
Asunto(s)
Cromatografía/métodos , Cilastatina/química , Inhibidores Enzimáticos/química , Imipenem/química , Espectrometría de Masas en Tándem/métodos , Inhibidores de beta-Lactamasas , Animales , Antibacterianos/sangre , Antibacterianos/química , Cilastatina/sangre , Inhibidores Enzimáticos/sangre , Haplorrinos , Imipenem/sangre , Ratones , RatasRESUMEN
A series of tricyclic CGRP receptor antagonists was optimized in order to improve oral bioavailability. Attenuation of polar surface area and incorporation of a weakly basic indoline nitrogen led to compound 5, a potent antagonist with good oral bioavailability in three species.
Asunto(s)
Antagonistas del Receptor Peptídico Relacionado con el Gen de la Calcitonina , Compuestos Heterocíclicos con 3 Anillos/farmacocinética , Compuestos de Espiro/farmacocinética , Administración Oral , Animales , Disponibilidad Biológica , Línea Celular , Perros , Haplorrinos , Compuestos Heterocíclicos con 3 Anillos/administración & dosificación , Compuestos Heterocíclicos con 3 Anillos/química , Humanos , Ratas , Receptores de Péptido Relacionado con el Gen de Calcitonina/metabolismo , Compuestos de Espiro/administración & dosificaciónRESUMEN
In our effort to find potent, orally bioavailable CGRP receptor antagonists for the treatment of migraine, a novel series based on a pyridinone template was investigated. After optimizing the privileged structure and the placement of the attached phenyl ring, systematic SAR was carried out on both the N-alkyl and C-5 aryl substituents. Several analogs with good potency and pharmacokinetic profiles were identified.
Asunto(s)
Antagonistas del Receptor Peptídico Relacionado con el Gen de la Calcitonina , Piridonas/farmacología , Administración Oral , Animales , Disponibilidad Biológica , Semivida , Piridonas/administración & dosificación , Piridonas/química , Piridonas/farmacocinética , Ratas , Relación Estructura-ActividadRESUMEN
Calcitonin gene-related peptide (CGRP) has been implicated in the pathogenesis of migraine. Herein we describe optimization of CGRP receptor antagonists based on an earlier lead structure containing a (3R)-amino-(6S)-phenylcaprolactam core. Replacement of the phenylimidazolinone with an azabenzimidazolone gave stable derivatives with lowered serum shifts. Extensive SAR studies of the C-6 aryl moiety revealed the potency-enhancing effect of the 2,3-difluorophenyl group, and trifluoroethylation of the N-1 amide position resulted in improved oral bioavailabilities, ultimately leading to clinical candidate 38 (MK-0974).
Asunto(s)
Azepinas/síntesis química , Antagonistas del Receptor Peptídico Relacionado con el Gen de la Calcitonina , Imidazoles/síntesis química , Trastornos Migrañosos/tratamiento farmacológico , Administración Oral , Animales , Azepinas/farmacocinética , Azepinas/farmacología , Disponibilidad Biológica , Células CACO-2 , Permeabilidad de la Membrana Celular , Perros , Humanos , Imidazoles/farmacocinética , Imidazoles/farmacología , Macaca mulatta , Ratas , Flujo Sanguíneo Regional/efectos de los fármacos , Piel/irrigación sanguínea , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
Thrombin-activatable fibrinolysis inhibitor (TAFI) is an important regulator of fibrinolysis, and inhibitors of this enzyme have potential use in antithrombotic and thrombolytic therapy. Appropriately substituted imidazole acetic acids such as 10j were found to be potent inhibitors of activated TAFI and selective versus the related carboxypeptidases CPA, CPN, and CPM but not CPB. Further, 10j accelerated clot lysis in vitro and was shown to be efficacious in a primate model of thrombosis.
Asunto(s)
Acetatos/síntesis química , Aminopiridinas/síntesis química , Carboxipeptidasa B2/antagonistas & inhibidores , Fibrinolíticos/síntesis química , Imidazoles/síntesis química , Propionatos/síntesis química , Inhibidores de Proteasas/síntesis química , Acetatos/farmacocinética , Acetatos/farmacología , Aminopiridinas/farmacocinética , Aminopiridinas/farmacología , Animales , Sitios de Unión , Carboxipeptidasa B2/química , Perros , Fibrinolíticos/farmacocinética , Fibrinolíticos/farmacología , Humanos , Imidazoles/farmacocinética , Imidazoles/farmacología , Técnicas In Vitro , Microsomas/metabolismo , Modelos Moleculares , Propionatos/farmacocinética , Propionatos/farmacología , Inhibidores de Proteasas/farmacocinética , Inhibidores de Proteasas/farmacología , Ratas , Relación Estructura-ActividadRESUMEN
A series of N-(1,3-thiazol-2-yl)pyridin-2-amine KDR kinase inhibitors have been developed that possess optimal properties. Compounds have been discovered that exhibit excellent in vivo potency. The particular challenges of overcoming hERG binding activity and QTc increases in vivo in addition to achieving good pharmacokinetics have been acomplished by discovering a unique class of amine substituents. These compounds have a favorable kinase selectivity profile that can be accentuated with appropriate substitution.
Asunto(s)
Aminopiridinas/síntesis química , Canales de Potasio con Entrada de Voltaje/metabolismo , Piridinas/síntesis química , Receptores de Factores de Crecimiento Endotelial Vascular/antagonistas & inhibidores , Tiazoles/síntesis química , Administración Oral , Aminopiridinas/farmacocinética , Aminopiridinas/farmacología , Animales , Disponibilidad Biológica , Línea Celular , Perros , Canal de Potasio ERG1 , Electrocardiografía/efectos de los fármacos , Canales de Potasio Éter-A-Go-Go , Técnicas In Vitro , Pulmón/enzimología , Macaca mulatta , Masculino , Ratones , Microsomas Hepáticos/metabolismo , Fosforilación , Piridinas/farmacocinética , Piridinas/farmacología , Ensayo de Unión Radioligante , Ratas , Ratas Sprague-Dawley , Tiazoles/farmacocinética , Tiazoles/farmacología , Distribución Tisular , Receptor 2 de Factores de Crecimiento Endotelial Vascular/metabolismoRESUMEN
INTRODUCTION: Telcagepant (MK-0974) is a novel, orally active and selective CGRP receptor antagonist being investigated for acute treatment of migraine. Early clinical data suggested greater than dose proportional increases in exposure following oral administration. The aim of the present studies was to definitively characterize the oral and IV dose proportionality of telcagepant. METHODS: Healthy adult subjects were enrolled in two separate open-label randomized dose proportionality studies: 1) single oral dose crossover from 50 to 600 mg (N = 19); 2) single IV dose parallel group from 5 to 250 mg (N = 10 per dose). Blood samples were collected at time points from 0 to 48 hours postdose. RESULTS: Telcagepant was rapidly absorbed with a T(max) of approximately 1 to 2 hours after oral administration. The terminal half-life was approximately 8 to 9 hours after IV dosing and approximately 4 to 7 hours after oral dosing. Oral administration of telcagepant resulted in greater than dose proportional increases in exposure, while IV administration resulted in approximately dose proportional increases in exposure. CONCLUSIONS: Telcagepant was generally well tolerated. Oral telcagepant exhibits non-linear pharmacokinetics.
RESUMEN
To eliminate the diastereomer interference on Telcagepant (MK-0974) determination during clinical study support, on-line high turbulent-flow liquid chromatography (HTLC) methods, HTLC-A and HTLC-B that covered dynamic range of 0.5-500 nM and 5-5000 nM, respectively, were developed. To meet the requirement of rapid assay transfer among multiple laboratories and analysts, a solid-phase extraction (SPE) assay was derived from the existing HTLC-B assay under the same dynamic range. The on-line HTLC assays were achieved through direct injection of plasma samples, extraction of analyte with a Cohesive C18 column (50 mm x 0.5 mm, 50 microm), followed by HPLC separation on a FluoPhase RP column (100 mm x 2.1 mm, 5 microm) and MS/MS detection. The off-line SPE assay used Waters Oasis HLB microElution plate to extract the analytes from plasma matrix before injecting on a FluoPhase RP column (150 mm x 2.1 mm, 5 microm) for LC-MS/MS analysis. Under both on-line and off-line assay conditions, the diastereomer 1c was chromatographically separated from MK-0974. Cross-validation with the pooled samples demonstrated that both on-line and off-line assays provided comparable data with a difference of < 2.6%. The assays were proved to be specific, accurate and reliable, and have been used to support multiple clinical studies. The pros and cons of on-line and off-line assays with regard to man power involved in sample preparation, total analysis time, carryover, cost efficiency, and the requirement for assay transfer are discussed.
Asunto(s)
Azepinas/sangre , Cromatografía Liquida/métodos , Análisis de Inyección de Flujo/métodos , Imidazoles/sangre , Espectrometría de Masas/métodos , Extracción en Fase Sólida/métodos , Espectrometría de Masas en Tándem/métodos , Azepinas/uso terapéutico , Humanos , Imidazoles/uso terapéutico , Trastornos Migrañosos/tratamiento farmacológicoRESUMEN
Telcagepant (MK-0974) is a novel oral calcitonin gene-related peptide (CGRP) receptor antagonist and is currently under clinical development. Results from phases II and III clinical trials have suggested that telcagepant is effective for migraine treatment. A reliable and high throughput protein precipitation (PPT) method for determination of telcagepant in human plasma using liquid chromatography coupled with atmospheric pressure chemical ionization (APCI) tandem mass spectrometry has been developed. Clinical samples, internal standard (IS) and acetonitrile are transferred into 96-well plates using a robotic liquid handling system. An aliquot of 10 microL supernatant is directly injected into the LC-MS/MS system where separation is performed on a FluoPhase RP (150 x 2.1mm, 5 microm) column with an isocratic mobile phase (60% acetonitrile with 0.1% formic acid and 40% water with 0.1% formic acid) at 0.2 mL/min. The interfering 3S-diastereomer of telcagepant, which is observed in clinical samples, is chromatographically resolved from telcagepant. The PPT procedure significantly reduces the time required for sample processing and the assay is sufficiently sensitive for detection using both API 4000 and API 3000 mass spectrometers. The linear calibration range is 5-5000 nM using 200 microL of plasma. Assay intraday validation was conducted using six calibration curves derived from six lots of human control plasma. Calibration standard accuracy did not deviate by more than 3% and 6% of nominal values, and precision did not exceed 4% coefficient of variation (CV) and 10% CV, respectively on the API 4000 and API 3000. Several clinical phases IIb and III studies have been successfully supported with this assay.
Asunto(s)
Azepinas/sangre , Cromatografía Líquida de Alta Presión/métodos , Imidazoles/sangre , Espectrometría de Masas en Tándem/métodos , Humanos , Trastornos Migrañosos/sangre , Trastornos Migrañosos/tratamiento farmacológicoRESUMEN
Pyrimidino-thiazolyl carbonitriles were prepared that are potent VEGFR-2 (KDR) kinase inhibitors. The modification of lead structures resulted in 3m which exhibited the best overall profile in KDR inhibitory activity, iv/po pharmacokinetics, and reduced hERG affinity.