Preparation of acetylmercapto-3-carboxypropanoyl insulins using preparative high-performance liquid chromatography on an anion-exchange column.
A method for the preparation of insulin derivatives which have protected sulfhydryl group(s) at definite site(s) on the molecule is described. Porcine insulin reacts with S-acetylmercaptosuccinic anhydride to afford four species of insulin derivatives that have 2 (or 3)-acetylmercapto-3-carboxypropanoyl group(s) at i) Gly(A1), ii) Gly(A1) and Phe(B1), iii) Gly(A1) and Lys(B29), and iv) Gly(A1), Phe(B1) and Lys(B29) positions. The derivatives are efficiently separated in a preparative scale by anion-exchange high-performance liquid chromatography on a TSKgel DEAE-2SW column. The four derivatives are all readily deacetylated with hydroxylamine to give the corresponding sulfhydryl insulin derivatives.
Nanami
M
M
Faculty of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
Zaitsu
K
K
Ohkura
Y
Y
D015097:3-Mercaptopropionic Acid / Q000031:analogs & derivatives* / Q000138:chemical synthesis; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D007328:Insulin / Q000031:analogs & derivatives* / Q000138:chemical synthesis; D013552:Swine
D007328:Insulin; D015097:3-Mercaptopropionic Acid