Use of cultured cerebral capillary endothelial cells in modeling the central nervous system availability of 2',3'-dideoxyinosine.
The biochemical and physiological mechanisms responsible for the limited central nervous system (CNS) uptake of dideoxynucleoside reverse transcriptase inhibitors currently used to treat HIV-1 infection in humans are poorly understood. In vitro models of the blood-brain barrier (BBB) offer an attractive alternative to in vivo or in situ animal studies for understanding the role of the blood-brain barrier in regulating brain tissue concentrations of these agents. In the present study, the kinetics of 2', 3'-dideoxyinosine (ddI) uptake and purine nucleoside phosphorylase (PNP) mediated catabolism in primary cultures of bovine brain microvessel endothelial cells (BBMECs) were determined in order to ascertain the importance of both transport and metabolism governing the CNS availability of this purine dideoxynucleoside. Initial rates of ddI uptake as a function of ddI donor concentration suggest the involvement of both passive diffusion and carrier-mediated processes. These studies confirm earlier in vivo findings that transporters may play a role in regulating the CNS concentration of ddI. Analysis of ddI uptake and metabolite accumulation in BBMECs over longer time intervals (beyond the intial rate region) provide substantial in vitro evidence for an enzymatic BBB for ddI. Simulations of the CNS availability of ddI derived from in vitro estimates of parameters for passive diffusion, carrier-mediation, and metabolism indicate that the fraction of ddI entering the BBB cells which actually reaches the brain parenchyma may be quite low (< 2%) due to metabolism by PNP localized within the BBB, consistent with the low CNS delivery of ddI observed in vivo. Transporters and metabolic enzymes within the BBB may function in coordinated fashion to reduce the CNS concentrations of both rapidly metabolized and poorly metabolized dideoxynucleosides.
Johnson
M D
MD
Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, Salt Lake City, Utah 84112, USA.
Anderson
B D
BD
D000818:Animals; D019380:Anti-HIV Agents / Q000493:pharmacokinetics*; D001812:Blood-Brain Barrier; D002196:Capillaries / Q000166:cytology / Q000378:metabolism; D002417:Cattle; D002469:Cell Separation; D002478:Cells, Cultured; D002490:Central Nervous System / Q000378:metabolism*; D002560:Cerebrovascular Circulation / Q000502:physiology*; D002851:Chromatography, High Pressure Liquid; D016049:Didanosine / Q000493:pharmacokinetics*; D004730:Endothelium, Vascular / Q000166:cytology* / Q000378:metabolism*; D007700:Kinetics; D008954:Models, Biological; D012044:Regression Analysis; D012995:Solubility; D013056:Spectrophotometry, Ultraviolet
D019380:Anti-HIV Agents; D016049:Didanosine