Transport of human recombinant brain-derived neurotrophic factor (BDNF) through the rat blood-brain barrier in vivo using vector-mediated peptide drug delivery.
The blood-brain barrier (BBB) transport of brain-derived neurotrophic factor (BDNF) in anesthetized rats was examined in the present studies using vector-mediated peptide drug delivery. Following tritiation, the BDNF was biotinylated via a disulfide linker and was coupled to a covalent conjugate of neutral avidin (NLA), which binds the biotinylated peptide with a high affinity, and the murine OX26 monoclonal antibody to the rat transferrin receptor. Owing to the abundance of transferrin receptors on brain capillary endothelium, the OX26 monoclonal antibody undergoes receptor-mediated transcytosis through the BBB, and the NLA-OX26 conjugate transports biotinylated peptide therapeutics through the BBB. The present studies show that while unconjugated BDNF was not transported through the BBB in vivo, the conjugation of biotinylated BDNF to the NLA-OX26 vector resulted in a marked increase in the brain delivery of BDNF, as defined by measurements of the percentage of the injected dose (ID) delivered per gram of brain. Although BDNF was not transported through the BBB in vivo, this cationic peptide was avidly bound by isolated human brain capillaries via a low-affinity, high-capacity system that was inhibited by protamine and by serum protein binding of BDNF. In conclusion, these studies show that the delivery of unconjugated BDNF to brain is nil owing to the combined effects of negligible BBB transport and rapid systemic clearance of intravenous administered BDNF. The brain delivery of BDNF may be augmented by conjugation of BDNF to BBB drug delivery vectors, such as the NLA-OX26 conjugate.
Pardridge
W M
WM
Department of Medicine, UCLA School of Medicine 90024.
Kang
Y S
YS
Buciak
J L
JL
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D001360:Avidin / Q000737:chemistry; D001682:Biological Availability; D001710:Biotin; D001812:Blood-Brain Barrier / Q000502:physiology; D019208:Brain-Derived Neurotrophic Factor; D002196:Capillaries / Q000378:metabolism; D002352:Carrier Proteins; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004220:Disulfides; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D009419:Nerve Tissue Proteins / Q000008:administration & dosage / Q000493:pharmacokinetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000493:pharmacokinetics
D000911:Antibodies, Monoclonal; D019208:Brain-Derived Neurotrophic Factor; D002352:Carrier Proteins; D004220:Disulfides; D009419:Nerve Tissue Proteins; D011994:Recombinant Proteins; D001360:Avidin; D001710:Biotin