In vivo cleavability of a disulfide-based chimeric opioid peptide in rat brain.
Brain delivery of systemically administered neuropeptide drugs may be achieved by the synthesis of chimeric peptides, wherein the peptide is coupled to transport vectors via avidin-biotin technology. The present study focuses on factors that optimize the linkage of drugs to transport vectors. The vector is the OX26 monoclonal antibody to the transferrin receptor, and the model peptide used in these studies is [Lys7]dermorphin (K7DA). The K7DA is monobiotinylated at the epsilon-amino group of the Lys7 residue with either a cleavable linker, e.g., disulfide, using NHS-SS-biotin, or a noncleavable linker, e.g., amide, using NHS-XX-biotin. Disulfide cleavage of the biotinylated derivative yields the desbiotinylated peptide, which is thiolated. Structures of the K7DA analogues were confirmed by secondary ion mass spectrometry. The biotinylated peptides were coupled to a thiol-ether conjugate of the OX26 antibody and either neutral avidin (NLA) or streptavidin. The binding constants (Ki) of the K7DA, the biotinylated K7DA (bio-XX-K7DA), the desbiotinylated K7DA, and the bio-XX-KD7A conjugated to NLA-OX26 were 0.62 +/- 0.14, 1.59 +/- 0.27, 1.24 +/- 0.24, and > 10 nM, respectively, and were determined with a mu-opioid peptide radioreceptor assay. Comparable results were obtained with in vivo tail-flick analgesia testing following intracerebroventricular (icv) injection of opioid chimeric peptides. Reversibility of pharmacologic action of thiolated peptide was demonstrated by icv naloxone administration. The cleavability of the disulfide linker in vivo in rat plasma and brain was assessed with gel filtration HPLC and internal carotid artery perfusion of labeled opioid chimeric peptides.(ABSTRACT TRUNCATED AT 250 WORDS)
Bickel
U
U
Department of Medicine, UCLA School of Medicine 90024, USA.
Kang
Y S
YS
Pardridge
W M
WM
D000700:Analgesics / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D001360:Avidin / Q000737:chemistry; D001710:Biotin / Q000737:chemistry; D001921:Brain / Q000378:metabolism*; D002339:Carotid Arteries / Q000187:drug effects / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D004220:Disulfides / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D007276:Injections, Intraventricular; D007457:Iodine Radioisotopes; D009270:Naloxone / Q000494:pharmacology; D009842:Oligopeptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D018847:Opioid Peptides / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D010147:Pain Measurement / Q000187:drug effects; D011869:Radioligand Assay; D051381:Rats; D017450:Receptors, Opioid, mu / Q000037:antagonists & inhibitors / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology
D000700:Analgesics; D004220:Disulfides; D007457:Iodine Radioisotopes; D009842:Oligopeptides; D018847:Opioid Peptides; D017450:Receptors, Opioid, mu; D011993:Recombinant Fusion Proteins; C091812:dermorphin, Lys(7)-; C063076:tyrosyl-arginyl-phenylalanyl-lysinamide; D001360:Avidin; D009270:Naloxone; D001710:Biotin