Loading patent…
AssigneeConselho Nacional de Desenvolvimento Cientifico E Technologico-CNPQ
InventorsClovis Ryuichi Nakaie, Simone Dos Reis Barbosa, Eduardo Maffud Cilli, Maria Teresa Lamy-Freund, Armando Siuiti Ito, Maria Aparecida Visconti, Ana Maria De Lauro Castrucci
Estimating the term…
The present invention refers to the chemical synthesis and potentiality for application in several biochemical assays, of a potent agonist of the α-melanocyte stimulating hormone (α-MSH), labeled with an amino acid-type paramagnetic spin probe (Toac, or 2,2,6,6-tetramethylpiperidine-1-oxyl-amino-4-carboxylic acid), valuable for use in electron spin resonance and fluorescence allowing biochemical-clinical investigation of relevant physiological roles of α-MSH in animal organism. The referred analogue of the present invention, acetil-Toac0-(Nle4, DPhe7)-α-MSH, where (Nle4, DPhe7)-α-(MSH is nominated commercially as Melanotan™ and although containing a non-natural compound in its structure such as Toac, maintained entirely its potent agonist potency. Its integral activity associated with the fact that it is naturally fluorescent (due to the Trp residue of its sequence) give to this labeled analogue a unique potential for a great variety of investigations regarding relevant physiological roles already known of this neuroimmuno-modulator.
“SYNTHESIS OF A POTENT PARAMAGNETIC AGONIST (EPM-3) OF THE α-MELANOCYTE STIMULATING HORMONE CONTAINING AMINO ACID-TYPE STABLE FREE RADICAL”, characterized by presenting the chemical structure Acetil-Toac-Ser-Tyr-Ser-Nle-Glu-His-DPhe-Arg-Trp-Gly-Lys-Pro-Val-CONH2, denoted as Ac-Toac0-(Nle4,D-Phe7)-α-MSH and where Toac is the paramagnetic 2,2,6,6-tetramethylpiperidine-1-oxyl-4-amino-4-carboxylic acid, for application in any type of physical-chemical, biochemical or clinical assays of the α-melanocyte stimulating hormone (α-MSH).
“SYNTHESIS OF A POTENT PARAMAGNETIC AGONIST (EPM-3) OF THE α-MELANOCYTE STIMULATING HORMONE CONTAINING AMINO ACID-TYPE STABLE FREE RADICAL”, according to the preceding claims and characterized by the coupling of the paramagnetic probe Toac using its 9-fluorenylmethyloxycarbonyl (Fmoc)-derivative which allows the introduction of this spin probe internally to any peptide sequence followed by final acetylation step where a large amount of acetic anhydride in dimethylformamide was used for 1 hour.
“SYNTHESIS OF A POTENT PARAMAGNETIC AGONIST (EPM-3) OF THE α-MELANOCYTE STIMULATING HORMONE CONTAINING AMINO ACID-TYPE STABLE FREE RADICAL”, according to the preceding claims, wherein the synthesized peptide was cleavage from the resin with anhydrous HF for 90 min at 0° C., being added the ethanedithiol for the removal of the formyl-protecting group of the tryptophan residue and of o-cresol and dimethylsulfide to minimize side reactions during this treatment in strong anhydrous acid with the proportion HF:o-cresol:dimethylsulfide:ethanedithiol (8.5:0.5:0,5:0.5), followed by resin washing with ethyl acetate and peptide extraction from the resin with 5% acetic acid in water followed by lyophilization.
Dependencies are inferred from the claim text.