title: Development of Cassette PAMPA for Permeability Screening.
abstract: The parallel artificial membrane permeability assay (PAMPA) is widely used in early-stage drug discovery to discriminate compounds by intestinal permeability. The purpose of the current study was to establish a cassette (n-in-1) PAMPA to enable permeability screening of lipophilic compounds. A double-sink PAMPA consisting of a pH gradient (i.e., pH 6.5 and 7.4 for the donor and receiver compartments, respectively) and a lipophilic sink (i.e., a surfactant in the receiver solution) was utilized with cassette incubation of 10 reference compounds. Sample analysis was conducted using selected reaction monitoring (SRM) with a triple quadrupole LC-MS/MS system. Correlation between PAMPA permeability and human intestinal absorption (HIA) of the reference compounds yielded two false negatives, namely propranolol (PPN) and verapamil (VER); these two compounds showed a substantially lower recovery (ca. 10%) than other reference compounds (>69%). This cassette PAMPA was repeated subsequently with polysorbate 80 added to the donor compartments, which resulted in a significant increase in both the recovery and the permeability of the false negatives. Accordingly, the permeability class of all reference compounds could be unambiguously differentiated using this cassette PAMPA. Also, a strong linear correlation (r=0.9845) was observed between the cassette and discrete permeability of all reference compounds.
authors.affiliation: College of Pharmacy, Korea University.
authors.forename: Mun Hwan
authors.initials: MH
authors.lastname: Oh
authors.forename: Hye Jin
authors.initials: HJ
authors.lastname: Lee
authors.forename: Sang Hyun
authors.initials: SH
authors.lastname: Jo
authors.forename: Bernie Byunghoon
authors.initials: BB
authors.lastname: Park
authors.forename: Sae-Bom
authors.initials: SB
authors.lastname: Park
authors.forename: Eun-Yeong
authors.initials: EY
authors.lastname: Kim
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Zhou
authors.forename: Young Ho
authors.initials: YH
authors.lastname: Jeon
authors.forename: Kiho
authors.initials: K
authors.lastname: Lee
mesh_terms: D018938:Caco-2 Cells; D002463:Cell Membrane Permeability / Q000187:drug effects / Q000502:physiology; D002626:Chemistry, Pharmaceutical / Q000379:methods*; D004353:Drug Evaluation, Preclinical / Q000379:methods; D005456:Fluorescent Dyes / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D007408:Intestinal Absorption / Q000187:drug effects / Q000502:physiology; D008567:Membranes, Artificial; D010539:Permeability / Q000187:drug effects*; D011136:Polysorbates / Q000378:metabolism* / Q000494:pharmacology; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D005456:Fluorescent Dyes; D008567:Membranes, Artificial; D011136:Polysorbates