The effect of beta-turn structure on the permeation of peptides across monolayers of bovine brain microvessel endothelial cells.
OBJECTIVE
To investigate the effects of the beta-turn structure of a peptide on its permeation via the paracellular and transcellular routes across cultured bovine brain microvessel endothelial cell (BBMEC) monolayers, an in vitro model of the blood-brain barrier (BBB).


METHODS
The effective permeability coefficients (Peff) of the model peptides were determined across BBMEC monolayers. The dimensions of the aqueous pores in the tight junctions (TJs) of the BBMEC monolayers were determined using a series of hydrophilic permeants. This value and the molecular radius of each peptide were used to calculate the theoretical paracellular (PP*) and transcellular (PT*) permeability coefficients for each peptide.


RESULTS
A comparison of the theoretical PP* values with the observed Peff values was made for a series of model peptides. For the most hydrophobic peptides (Ac-PheProXaaIle-NH2 and Ac-PheProXaaIleVal-NH2; Xaa = Gly, Ile), it was concluded that the Gly-containing peptide of each pair more readily permeates BBMEC monolayers via the transcellular pathway than the Ile-containing analog. In addition, the Gly-containing peptides, which exhibit more beta-turn structure, were shown to be more lipophilic than the Ile-containing peptides as estimated by the log of their 1-octanol:HBSS partition coefficients (log Po/w). However, the three hydrophilic peptide pairs (Ac-TyrProXaaAspVal-NH2, Ac-TyrProXaaAsnVal-NH2, and Ac-TyrProXaaIleVal-NH2; Xaa = Gly, Ile) were found to permeate BBMEC monolayers predominantly via the paracellular pathway. No differences were observed in the Peff values of the hydrophilic peptides having higher beta-turn structures as compared to the peptides lacking these structural features. In addition, the Ile-containing peptides exhibited significantly higher log Po/w values than the Gly-containing hydrophilic peptides.


CONCLUSIONS
Hydrophobic peptides that exhibit significant beta-turn structure in solution are more lipophilic as measured by log Po/w, and more readily permeate BBMEC monolayers via the transcellular route than hydrophobic peptides that lack this type of solution structure. Similar secondary structural features in hydrophilic peptides do not appear to sufficiently alter the physicochemical properties of the peptides so as to alter their paracellular flux through BBMEC monolayers.
Sorensen
M
M
Department of Pharmaceutics, Royal Danish School of Pharmacy, Copenhagen, Denmark.
Steenberg
B
B
Knipp
G T
GT
Wang
W
W
Steffansen
B
B
Frokjaer
S
S
Borchardt
R T
RT
D000818:Animals; D001812:Blood-Brain Barrier; D001921:Brain / Q000098:blood supply*; D002417:Cattle; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D008833:Microcirculation; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism*; D010539:Permeability; D017433:Protein Structure, Secondary; D012996:Solutions; D013329:Structure-Activity Relationship; D019108:Tight Junctions / Q000378:metabolism
D009842:Oligopeptides; D012996:Solutions