title: Metabolism-based brain-targeting system for a thyrotropin-releasing hormone analogue.
abstract: Gln-Leu-Pro-Gly, a progenitor sequence for the thyrotropin-releasing hormone (TRH) analogue [Leu(2)]TRH (pGlu-Leu-Pro-NH(2)), was covalently and bioreversibly modified on its N- and C-termini (by a 1,4-dihydrotrigonellyl and a cholesteryl group, respectively) to create lipoidal brain-targeting systems for the TRH analogue. The mechanism of targeting and the recovery of the parent peptide at the target site involve several enzymatic steps, including the oxidation of the 1,4-dihydropyridine moiety. Due to the lipid insolublity of the peptide pyridinium conjugate obtained after this reaction, one of the rudimentary steps of brain targeting (i.e., trapping in the central nervous system) can be accomplished. Our design also included spacer amino acid(s) inserted between the N-terminal residue of the progenitor sequence and the dihydrotrigonellyl group to facilitate the posttargeting removal of the attached modification. The release of the TRH analogue in the brain is orchestrated by a sequential metabolism utilizing esterase/lipase, peptidyl glycine alpha-amidating monooxygenase (PAM), peptidase cleavage, and glutaminyl cyclase. In addition to in vitro experiments to prove the designed mechanism of action, the efficacy of brain targeting for [Leu(2)]TRH administered in the form of chemical-targeting systems containing the embedded progenitor sequence was monitored by the antagonistic effect of the peptide on the barbiturate-induced anesthesia (measure of the activational effect on cholinergic neurons) in mice, and considerable improvement was achieved over the efficacy of the parent peptide upon using this paradigm.
authors.affiliation: Center for Drug Discovery, College of Pharmacy, University of Florida, Gainesville, Florida 32610-0497, USA.
authors.forename: L
authors.initials: L
authors.lastname: Prokai
authors.forename: K
authors.initials: K
authors.lastname: Prokai-Tatrai
authors.forename: X
authors.initials: X
authors.lastname: Ouyang
authors.forename: H S
authors.initials: HS
authors.lastname: Kim
authors.forename: W M
authors.initials: WM
authors.lastname: Wu
authors.forename: A
authors.initials: A
authors.lastname: Zharikova
authors.forename: N
authors.initials: N
authors.lastname: Bodor
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism; D000697:Central Nervous System Stimulants / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D015195:Drug Design; D066298:In Vitro Techniques; D051379:Mice; D011355:Prodrugs / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D011761:Pyrrolidonecarboxylic Acid / Q000031:analogs & derivatives; D013973:Thyrotropin-Releasing Hormone / Q000031:analogs & derivatives* / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism
chemical_list: D000697:Central Nervous System Stimulants; D011355:Prodrugs; C401384:pyroglutamyl-leucyl-prolinamide; D013973:Thyrotropin-Releasing Hormone; D011761:Pyrrolidonecarboxylic Acid