Synthesis of human beta-endorphin in solution using benzyl-type side chain protective groups.
A solution synthesis of human beta-endorphin (beta-EP) was carried out by condensation of protected peptide segments bearing N alpha-tert.-butyloxycarbonyl groups and benzyl-derived groups for the protection of functionalities in amino acid side chains. Five intermediate segments were assembled in a stepwise manner starting at the carboxyl terminus. Thus, the segment of sequence region (27-31) was coupled to segment (22-26) by the azide method. Segment (19-21) was incorporated into the growing chain by azide coupling, and segment (10-18) by dicyclohexylcarbodiimide coupling in the presence of 1-hydroxybenzotriazole (DDC-HOBt). Solubility problems in condensing the ensuing 22-peptide with segment (1-9) by DDC-HOBt were overcome by using a dimethylformamidephenol mixture as a solvent. Protecting group cleavage by Na in liquid NH3 was much superior to liquid HF which gave rise to many decomposition products. Homogeneous betah-EP indistinguishable from authentic material in physiochemical and biological properties, was obtained in a single preparative reversed phase liquid chromatographic step after protecting group cleavage.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001593:Benzyl Compounds; D002942:Circular Dichroism; D004723:Endorphins / Q000138:chemical synthesis* / Q000302:isolation & purification / Q000494:pharmacology; D006168:Guinea Pigs; D001615:beta-Endorphin
D000596:Amino Acids; D001593:Benzyl Compounds; D004723:Endorphins; D001615:beta-Endorphin