Ys. Kang et al., Stability of the disulfide bond in an avidin-biotin linked chimeric peptide during in vivo transcytosis through brain endothelial cells, J DRUG TAR, 8(6), 2000, pp. 425-434
Drug delivery of potential neuropharmaceuticals with poor intrinsic permeab
ility through the blood-brain barrier (BBB). such as peptides, is facilitat
ed by coupling to a vector that undergoes receptor-mediated transcytosis th
rough the endothelial cells of brain microvessels. When cleavable disulfide
linkers are used in the synthesis of such "chimeric peptides", it is cruci
al that the S-S-bridge is stable during transcytosis. Cleavage within endot
helial cells could result in sequestration of the drug moiety instead of pa
ssage through the BBB. In the present study the metabolically stable opioid
peptide [H-3]DALDA ([H-3]Tyr-DArg-Phe-Lys-NH2) was used as a model drug. I
t was monobiotinylated with the cleavable biotin reagent sulfosuccinimidyl
2-(biotinamido)ethyl-1,3'-dithiopropionate (NHS-SS-biotin) to obtain bio-[H
-3]DALDA. The biotinylated peptide was then bound to a vector for brain del
ivery after intravenous injection in rats, a covalent conjugate of streptav
idin and the transferrin receptor monoclonal antibody, OX26. Compared to pe
ptide without vector, brain uptake of bio-[H-3]DALDA after was increased 18
-fold to reach 0.12% of the injected dose per g tissue. Transcranial microd
ialysis was performed for 60 min after an intravenous bolus of chimeric pep
tide. followed by reverse phase HPLC of dialysate. Stability of the chimeri
c peptide during transport through the BBB into brain extracellular fluid w
as concluded from the absence of a peptide peak generated by disulfide clea
vage.