A practical reagent for reversible amino-protection of insulin, 3,4,5,6-tetrahydrophthalic anhydride.
Of the examined dicarboxylic acid anhydrides, 3,4,5,6-tetrahydrophthalic anhydride was found to be the best reagent in practical use for reversible amino-protection of insulin in terms of the rapidity of acid-deprotection. Twelve Gly(A1),Phe(B1),Lys(B29)-triacyl-insulins were prepared by reaction of porcine insulin with the dicarboxylic acid anhydrides and time courses for the deprotection of the acylated insulins with dilute acetic acid were investigated by means of capillary zone electrophoresis, and the tetrahydrophthalyl(THP)-insulin obtained with the reagent was the most rapidly deacylated (6 h, 25 degrees C, 0.1 M acetic acid). Isolation of triacyl-insulins and Gly(A1)-THP-, Gly(A1),Phe(B1)-diTHP- and Gly(A1),Lys(B29)-diTHP-insulins using DEAE anion-exchange high-performance liquid chromatography is also described.
Kimura
Y
Y
Faculty of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
Zaitsu
K
K
Motomura
Y
Y
Ohkura
Y
Y
D002851:Chromatography, High Pressure Liquid; D007202:Indicators and Reagents; D007328:Insulin; D010796:Phthalic Anhydrides; D013696:Temperature
D007202:Indicators and Reagents; D007328:Insulin; D010796:Phthalic Anhydrides