title: Evaluation of in vitro brain penetration: optimized PAMPA and MDCKII-MDR1 assay comparison.
abstract: Parallel artificial membrane permeability assay (PAMPA) is arising in ADMET screening as a powerful tool to determine the passive permeability of new potential chemical entities. In an attempt to set up a sensitive high throughput method to assess passive blood-brain barrier (BBB) penetration we focused our attention on the effect of solvent and the influence of phospholipids on the permeability in PAMPA. Moreover, the high throughput nature of the assay was maximized by decreasing the incubation time and performing the assay in a cassette mode. UPLC system coupled with a mass spectrometer enormously reduces the analytical time, contemporaneously increasing the sensitivity of the method. P(app) values obtained from PAMPA were compared to permeability values from MDCKII-MDR1 assay. Evaluation of the two in vitro models with in vivo data was performed to test the predicting capacity of the two methods. Their contemporary assessment was shown to be an helpful tool in understanding the prevalent mechanism of penetration through the BBB.
authors.affiliation: Analytical and Developability Department, NiKem Research S.r.l., Via Zambeletti 25, 20021 Baranzate, Milan, Italy.
authors.forename: Simone
authors.initials: S
authors.lastname: Carrara
authors.forename: Veronica
authors.initials: V
authors.lastname: Reali
authors.forename: Paola
authors.initials: P
authors.lastname: Misiano
authors.forename: Giulio
authors.initials: G
authors.lastname: Dondio
authors.forename: Chiara
authors.initials: C
authors.lastname: Bigogno
mesh_terms: D001812:Blood-Brain Barrier / Q000187:drug effects / Q000378:metabolism*; D001921:Brain / Q000378:metabolism*; D002460:Cell Line; D002463:Cell Membrane Permeability; D002851:Chromatography, High Pressure Liquid; D003627:Data Interpretation, Statistical; D005079:Excipients; D006801:Humans; D013058:Mass Spectrometry; D008567:Membranes, Artificial; D010539:Permeability; D004364:Pharmaceutical Preparations / Q000378:metabolism; D010743:Phospholipids / Q000737:chemistry / Q000494:pharmacology; D015203:Reproducibility of Results; D012996:Solutions; D012997:Solvents
chemical_list: D005079:Excipients; D008567:Membranes, Artificial; D004364:Pharmaceutical Preparations; D010743:Phospholipids; D012996:Solutions; D012997:Solvents