In vitro model for predicting the access and distribution of drugs in the brain using hCMEC/D3 cells

Abstract

The BBB is a protective entity that prevents external substances from reaching the CNS but it also hinders the delivery of drugs into the brain when they are needed. The main objective of this work was to improve a previously proposed in vitro cell-based model by using a more physiological cell line (hCMEC/D3) to predict the main pharmacokinetic parameters that describe the access and distribution of drugs in the CNS: Kp_uu,brain, f_u,plasma, f_u,brain and V_u,brain. The hCMEC/D3 permeability of seven drugs was studied in transwell systems under different conditions (standard, modified with albumin and modified with brain homogenate). From the permeability coefficients of those experiments, the parameters mentioned above were calculated and four linear IVIVCs were established. The best ones were those that relate the in vitro and in vivo V_u,brain and f_u,brain (r² = 0.961 and r² = 0.940) which represent the binding rate of a substance to the brain tissue, evidencing the importance of using brain homogenate to mimic brain tissue when an in vitro brain permeability assay is done. This methodology could be a high-throughput screening tool in drug development to select the CNS promising drugs in three different in vitro BBB models (hCMEC/D3, MDCK and MDCK-MDR1).

Keywords: Blood−brain barrier (BBB); Distribution volume in brain (V(u,brain)); IVIVC; Plasma−brain partition coefficient (Kp(uu,brain)); Unbound fraction (f(u)).