Ishiguro_2014_Drug.Metab.Dispos_42_250

Reference

Title : Impact of endogenous esterase activity on in vitro p-glycoprotein profiling of dabigatran etexilate in caco-2 monolayers - Ishiguro_2014_Drug.Metab.Dispos_42_250
Author(s) : Ishiguro N , Kishimoto W , Volz A , Ludwig-Schwellinger E , Ebner T , Schaefer O
Ref : Drug Metabolism & Disposition: The Biological Fate of Chemicals , 42 :250 , 2014
Abstract :

Dabigatran etexilate, a double prodrug of dabigatran, is a reversible, competitive, direct thrombin inhibitor that has been approved for use in many countries. A recent guideline from the European Medicines Agency on drug-drug interactions proposed dabigatran etexilate as a sensitive in vivo and in vitro probe substrate for intestinal P-glycoprotein (P-gp) inhibition. We therefore performed a series of in vitro studies to determine the best experimental conditions for evaluation of P-gp involvement on the transport process of dabigatran etexilate across colorectal adenocarcinoma Caco-2 cell monolayers. Experiments using expressed carboxylesterase 1 (CES1) and CES2 bactosomes revealed that dabigatran etexilate was hydrolyzed into BIBR 1087 by CES1 expressed in our Caco-2 cells. The impact of CES1-mediated BIBR 1087 formation during transcellular transport experiments was assessed by comparing several combinations of three experimental approaches: radioactivity detection using [(14)C]dabigatran etexilate as substrate, liquid chromatography-tandem mass spectrometry (LC-MS/MS) quantification of dabigatran etexilate, and in the presence and absence of a CES inhibitor bis(p-nitrophenyl) phosphate (BNPP). The experimental approach that was based on the use of nonlabeled dabigatran etexilate together with LC-MS/MS quantification and the addition of BNPP was selected as the most favorable condition in which to correctly evaluate the permeability coefficient (Papp) of dabigatran etexilate and its transcellular transport by P-gp. The in vitro Caco-2 study at the selected condition revealed that dabigatran etexilate is a P-gp substrate with an efflux ratio of 13.8 and an intrinsic Papp, which is the Papp under the condition of complete blockage of P-gp by P-gp inhibitor, of 29 x 10(-6) cm/s.

PubMedSearch : Ishiguro_2014_Drug.Metab.Dispos_42_250
PubMedID: 24212377

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Citations formats

Ishiguro N, Kishimoto W, Volz A, Ludwig-Schwellinger E, Ebner T, Schaefer O (2014)
Impact of endogenous esterase activity on in vitro p-glycoprotein profiling of dabigatran etexilate in caco-2 monolayers
Drug Metabolism & Disposition: The Biological Fate of Chemicals 42 :250

Ishiguro N, Kishimoto W, Volz A, Ludwig-Schwellinger E, Ebner T, Schaefer O (2014)
Drug Metabolism & Disposition: The Biological Fate of Chemicals 42 :250