title: Modification of a PAMPA model to predict passive gastrointestinal absorption and plasma protein binding.
abstract: The Parallel Artificial Membrane Permeability Assay (PAMPA) is a well-known high throughput screening (HTS) technique for predicting in vivo passive absorption. In this technique, two compartments are separated by an artificial membrane that mimics passive permeability through biological membranes such as the dermal layer, the gastrointestinal tract (GIT), and the blood brain barrier (BBB). In the present study, a hexadecane artificial membrane (HDM)-PAMPA was used to predict the binding of compounds towards the human plasma using a mixture of human serum albumin (HSA) and alpha-1-acid glycoprotein (AGP). The ratio of HSA and AGP was equivalent to that found in the human plasma for both proteins (∼20:1). A pH gradient (5.0-7.4) was performed to increase the screening capacity and overcome the issue of passive permeability for acidic and amphoteric compounds. With this assay, the prediction of passive GIT absorption was maintained and the compounds were discriminated according to their permeability (on a no-to-high scale). The plasma protein binding (PPB) was estimated via the correlation of the differences between the amount of compound crossing the artificial membrane in assays conducted with and without protein using only a two end-point measurement. The use of a mixture of HSA and AGP to modulate drug permeation was compared to the use of the same concentrations of HSA and AGP used separately. The addition of HSA alone in the acceptor compartment was sufficient for estimating PPB, while it was demonstrated that AGP alone could enable the estimation of AGP binding.
authors.affiliation: School of Pharmaceutical Sciences, EPGL, University of Geneva, 30 Quai Ernest Ansermet, CH 1211 Geneva 4, Switzerland.
authors.forename: Alban
authors.initials: A
authors.lastname: Bujard
authors.affiliation: School of Pharmaceutical Sciences, EPGL, University of Geneva, 30 Quai Ernest Ansermet, CH 1211 Geneva 4, Switzerland.
authors.forename: Hervé
authors.initials: H
authors.lastname: Voirol
authors.affiliation: School of Pharmaceutical Sciences, EPGL, University of Geneva, 30 Quai Ernest Ansermet, CH 1211 Geneva 4, Switzerland.
authors.forename: Pierre-Alain
authors.initials: PA
authors.lastname: Carrupt
authors.affiliation: School of Pharmaceutical Sciences, EPGL, University of Geneva, 30 Quai Ernest Ansermet, CH 1211 Geneva 4, Switzerland. Electronic address: julie.schappler@unige.ch.
authors.forename: Julie
authors.initials: J
authors.lastname: Schappler
mesh_terms: D001798:Blood Proteins / Q000378:metabolism*; D001812:Blood-Brain Barrier; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007408:Intestinal Absorption; D008954:Models, Biological; D011485:Protein Binding; D013056:Spectrophotometry, Ultraviolet
chemical_list: D001798:Blood Proteins
keywords: AGP; Alpha-1-acid glycoprotein; HSA; Human serum albumin; PAMPA; PPB; Passive permeability; Plasma protein binding