Validation of an Ex Vivo Permeation Method for the Intestinal Permeability of Different BCS Drugs and Its Correlation with Caco-2 In Vitro Experiments

Abstract

The absorption study of drugs through different biological membranes constitutes an essential step in the development of new pharmaceutical dosage forms. Concerning orally administered forms, methods based on monolayer cell culture of Caco-2 (Caucasian colon adenocarcinoma) have been developed to emulate intestinal mucosa in permeability studies. Although it is widely accepted, it has disadvantages, such as high costs or high technical complexity, and limitations related to the simplified structure of the monolayer or the class of molecules that can be permeated according to the transport mechanisms. The aim of this work was to develop a new ex vivo methodology which allows the evaluation of the intestinal apparent permeability coefficient (Papp) while using fewer resources and to assess the correlation with Caco-2. To this end, pig (Sus scrofa) duodenum segments were mounted in Franz diffusion cells and used to permeate four different drugs: ketorolac tromethamine (Kt), melatonin (Mel), hydrochlorothiazide (Htz), and furosemide (Fur). No statistically significant differences (p > 0.05) were observed corelating Papp values from Franz diffusion cells and Caco-2 cell experiments for Kt, Htz, and Fur. However, there were statistically significant differences (p < 0.05) correlating Papp values and Mel. The difference is explained by the role of Mel in the duodenal epithelial paracellular permeability reduction. Ex vivo permeation may be an equivalent method to Caco-2 for drugs that do not produce intestinal membrane phenomena that could affect absorption.

Study of Melatonin as Preventive Agent of Gastrointestinal Damage Induced by Sodium Diclofenac

“…As the previous stage of in vivo studies, we set and validated an ex vivo model [54], which allows the study of the local effect of SD and MLT in the small intestinal mucosa; at the same time, the model allows the evaluation of the intestinal apparent permeability coefficient (Papp) for SD. Papp of SD alone and in combination with MLT were compared to evaluate the influence of MLT in the intestinal absorption of SD.…”

“…Ex vivo experiments were performed as described a previous paper [54] in the duodenum, the most proximal portion of the small intestine, of young female pigs (Sus scrofa). Animals were sacrificed for other purposes in the Animal Facility at Bellvitge Campus (University of Barcelona, Barcelona, Spain) and intestinal samples were obtained according to the 3R (reduction, refinement and replacement) principle.…”

Transport Mechanisms of Polymannuronic Acid and Polyguluronic Acid Across Caco-2 Cell Monolayers

“…The Caco-2 cell monolayer model is widely used to mimic the absorption and transport of drugs across the intestinal epithelial layer [13]. In this paper, we used the method of Sevin et al to establish a fast 7 day Caco-2 cell monolayer model to save modeling time.…”

Development of a Hierarchical Support Vector Regression-Based In Silico Model for Caco-2 Permeability

“…Moreover, Caco-2, which can be adopted to evaluate the drug permeability through the cytoplasm (transcellular uptake) or between cells (paracellular uptake) and active transport [ 6 ], has become the golden standard for predicting intestinal drug permeability and absorption because of its similarity in morphology and function with human enterocytes [ 21 , 22 , 23 ]. The Caco-2 protocol has been clearly described in detail by Hubatsch et al As compared with the biological membrane, the Caco-2 system still suffers from a range of disadvantages such as high technical complexity, the limitations related to the differences between cell monolayers and intestinal membrane structurally and functionally [ 24 ], in addition to its long culture periods (21‒24 days) with the significantly extensive costs, contributing to the major concerns in practical applications [ 21 , 25 ].…”