title: A high-throughput cell-based method to predict the unbound drug fraction in the brain.
abstract: Optimization of drug efficacy in the brain requires understanding of the local exposure to unbound drug at the site of action. This relies on measurements of the unbound drug fraction (fu,brain), which currently requires access to brain tissue. Here, we present a novel methodology using homogenates of cultured cells for rapid estimation of fu,brain. In our setup, drug binding to human embryonic kidney cell (HEK293) homogenate was measured in a small-scale dialysis apparatus. To increase throughput, we combined drugs into cassettes for simultaneous measurement of multiple compounds. Our method estimated fu,brain with an average error of 1.9-fold. We propose that our simple method can be used as an inexpensive, easily available and high-throughput alternative to brain tissues excised from laboratory animals. Thereby, estimates of unbound drug exposure can now be implemented at a much earlier stage of the drug discovery process, when molecular property changes are easier to make.
authors.affiliation: Department of Pharmacy, Uppsala University , SE-751 23 Uppsala, Sweden.
authors.forename: André
authors.initials: A
authors.lastname: Mateus
authors.forename: Pär
authors.initials: P
authors.lastname: Matsson
authors.forename: Per
authors.initials: P
authors.lastname: Artursson
mesh_terms: D000818:Animals; D001921:Brain / Q000187:drug effects* / Q000378:metabolism*; D003956:Dialysis; D057809:HEK293 Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D004364:Pharmaceutical Preparations / Q000032:analysis* / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D004364:Pharmaceutical Preparations