Asunto(s)
Antineoplásicos/uso terapéutico , Drogas en Investigación/uso terapéutico , Oncología Médica , Neoplasias/tratamiento farmacológico , Antineoplásicos/administración & dosificación , Antineoplásicos/química , Antineoplásicos/farmacología , Ensayos Clínicos como Asunto , Drogas en Investigación/administración & dosificación , Drogas en Investigación/química , Drogas en Investigación/farmacología , Humanos , Estructura MolecularAsunto(s)
Arritmias Cardíacas/inducido químicamente , Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos , Electrocardiografía/métodos , Animales , Arritmias Cardíacas/diagnóstico , Evaluación Preclínica de Medicamentos/métodos , Electrofisiología/métodos , Guías como Asunto , Humanos , Síndrome de QT Prolongado/inducido químicamente , Síndrome de QT Prolongado/diagnósticoAsunto(s)
Farmacología/tendencias , Animales , Antiinflamatorios/farmacología , Antineoplásicos/farmacología , Antivirales/farmacología , Enfermedades Autoinmunes/tratamiento farmacológico , Enfermedades del Sistema Nervioso Central/tratamiento farmacológico , Industria Farmacéutica , Receptores ErbB/antagonistas & inhibidores , Inhibidores de Integrasa VIH/farmacología , Humanos , Enfermedades Metabólicas/tratamiento farmacológico , Enfermedades del Sistema Nervioso/tratamiento farmacológico , InvestigaciónRESUMEN
PURPOSE: This study used a Box-Behnken experimental design to optimise the experimental conditions in the Caco-2 assay for a series of p-hydroxybenzoate ester compounds (log P 1.96-5.69), as highly lipophilic compounds are not handled well in this system. METHODS: Caco-2 cells, passage 55-70, were cultured on Transwelltrade mark cell culture supports and permeability assays were performed on day 21. A three level three factorial experimental design was used to optimise the experimental conditions. RESULTS: Addition of BSA (4% w/v) in the medium increased the apparent permeability coefficients (Papp) of each of the parabens except the octyl ester. Increasing the stirring rate by 100 rpm increased Papp for all the parabens. Use of simulated intestinal fluid either increased (fasted state) or decreased (fed state) the Papp of methyl-butyl parabens. CONCLUSIONS: The optimised conditions were; 1.55% w/v BSA, 215 rpm stirring rate and 3.02 mM sodium taurocholate in the simulated intestinal fluid; where octyl paraben (log P 5.69) had an Papp of 33.93 +/- 1.84 x 10(-6) cm/s, reflecting its rapid absorption in man. This study provides a systematic optimisation of the Caco-2 permeability assay to avoid the underestimation of the intestinal permeability of compounds with log P > 3.
Asunto(s)
Células CACO-2/fisiología , Absorción Intestinal/fisiología , Algoritmos , Líquidos Corporales/fisiología , Química Farmacéutica , Cromatografía Líquida de Alta Presión , Humanos , Lípidos/química , Parabenos/análisis , Parabenos/farmacocinética , Permeabilidad , Albúmina Sérica Bovina/químicaRESUMEN
Noncellular and cellular in vitro models for predicting intestinal absorption were used to investigate the transport and metabolism of parabens. The biomimetic artificial membrane permeability assay (BAMPA) membrane was constructed by impregnating a lipid solution on a hydrophobic filter. Caco-2 cells at passage numbers 65 to 80 were cultured in either the accelerated 3-day Biocoat system or the standard 21-day Transwell cell culture system. Paraben transport across the BAMPA system showed a parabolic relationship. The lowest log P (p-hydroxybenzoic acid) and highest log P compounds (heptyl and octyl parabens) had apparent permeabilities (Papp) less than 1.0 x 10(-6) cm/s and Papp was maximal at approximately 8.5 x 10(-6)cm/s for the intermediate log P (ethylparaben) compound. With the Biocoat, a similar parabolic relationship was found. In the 21-day Caco-2 cells, the parabens were metabolized by esterases at to p-hydroxybenzoic acid. In conclusion, the in vitro models added complementary insight into the absorption process, such as the transport route, intrinsic permeability, and extent of metabolism of the parabens. This study indicated that presystemic metabolism of orally ingested parabens to the p-hydroxybenzoic acid in the intestine may limit systemic exposure to alkyl-paraben esters in vivo.