In Vitro Intrinsic Permeability: A Transporter-Independent Measure of Caco-2 Cell Permeability in Drug Design and Development.
In vitro permeability data have a central place in absorption risk assessments in drug discovery and development. For compounds where active efflux impacts permeability in vitro, the inherent passive membrane permeability ("intrinsic permeability") gives a concentration-independent measure of the compound's permeability. This work describes the validation of an in vitro intrinsic permeability assay and application of the data in a predictive in silico model. Apparent intrinsic permeability (Papp) across Caco-2 cell monolayers is determined in the presence of an optimized cocktail of chemical inhibitors toward the three major efflux transporters ABCB1, ABCC2, and ABCG2. The intrinsic Pappvalue gives an estimate of passive permeability, which is independent of transporter expression levels and not limited by solubility or cell toxicity. An in silico model has been established to predict the Caco-2 intrinsic permeability and shown to consistently identify highly permeable compounds. The new intrinsic permeability assay is useful for early absorption estimates and suitable for absorption risk assessment in DMPK and pharmaceutical development.
Molecular Screening and Profiling, Discovery Sciences, ‡Predictive Compound ADME and Safety, Discovery Safety, Drug Safety and Metabolism, and §ADME and Biotransformation, DMPK Cardiovascular and Metabolic Diseases, Innovative Medicines Biotech Unit, AstraZeneca R&D Gothenburg , Mölndal 431 83, Sweden.
Linda
0000-0002-2104-5617
L
Fredlund
Molecular Screening and Profiling, Discovery Sciences, ‡Predictive Compound ADME and Safety, Discovery Safety, Drug Safety and Metabolism, and §ADME and Biotransformation, DMPK Cardiovascular and Metabolic Diseases, Innovative Medicines Biotech Unit, AstraZeneca R&D Gothenburg , Mölndal 431 83, Sweden.
Susanne
S
Winiwarter
Molecular Screening and Profiling, Discovery Sciences, ‡Predictive Compound ADME and Safety, Discovery Safety, Drug Safety and Metabolism, and §ADME and Biotransformation, DMPK Cardiovascular and Metabolic Diseases, Innovative Medicines Biotech Unit, AstraZeneca R&D Gothenburg , Mölndal 431 83, Sweden.
Constanze
C
Hilgendorf
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2 / Q000378:metabolism; D001693:Biological Transport, Active / Q000502:physiology; D018938:Caco-2 Cells; D015195:Drug Design; D055808:Drug Discovery; D006801:Humans; D007408:Intestinal Absorption / Q000502:physiology; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000378:metabolism
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins; C000715087:ABCC2 protein, human
Caco-2; active transport; drug discovery; in silico modeling; in vitro−in vivo correlation; intestinal absorption; intrinsic permeability; passive diffusion