In-vitro characterization of blood-brain barrier permeability to delta sleep-inducing peptide.
The diffusion of delta sleep-inducing peptide (DSIP) across the blood-brain barrier (BBB) has been investigated with an in-vitro model comprised of primary cultures of brain microvessel endothelial cell (BMEC) monolayers. The BMEC monolayers were mounted in a side-by-side diffusion apparatus and the transendothelial flux of DSIP analysed by HPLC with UV detection at 280 nm. The transendothelial flux of the peptide was linear with time and increasing concentrations of DSIP (non-saturable), but was not altered by reduced temperature. The apparent permeability coefficient for DSIP penetration of BMEC monolayers was in a range similar to water-soluble substances (e.g. fluorescein, fluorescein isothiocyanate dextrans) that penetrate the blood-brain barrier to a limited degree based on molecular weight. DSIP flux across the BMEC monolayers was also found to be bidirectional, insensitive to metabolic inhibitors, and not altered by high concentrations of tryptophan. Little degradation (apparent t1/2 about 10 h) of DSIP to major metabolites, tryptophan (trp) and des-trp DSIP, occurred over the time of the diffusion experiments. The results of these studies support and confirm observations in-vivo indicating that intact DSIP crosses the BBB by simple transmembrane diffusion.
Raeissi
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Department of Pharmaceutical Chemistry, University of Kansas, Lawrence 66045.
Audus
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KL
D000818:Animals; D001812:Blood-Brain Barrier; D002417:Cattle; D002478:Cells, Cultured; D003701:Delta Sleep-Inducing Peptide / Q000493:pharmacokinetics*; D004058:Diffusion; D066298:In Vitro Techniques; D008567:Membranes, Artificial; D008970:Molecular Weight; D010539:Permeability; D013696:Temperature; D014364:Tryptophan / Q000493:pharmacokinetics
D008567:Membranes, Artificial; D003701:Delta Sleep-Inducing Peptide; D014364:Tryptophan