Preparation of S-acetylthioglycoloyl insulins based on separation by anion-exchange high-performance liquid chromatography.
A method for the preparation of insulin derivatives having protected sulfhydryl group(s) on definite site(s) on the molecule which uses anion-exchange high performance liquid chromatography on a TSKgel DEAE-2SW column for separation is described. Porcine insulin reacts with N-succinimidyl S-acetylthioacetate to afford four species of insulin derivatives that have S-acetylthioglycoloyl 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. An insulin derivative which has a group at the Lys(B29)-position is prepared by the S-acetylthioglycoloylation of Gly(A1), Phe(B1)-dicitraconyl insulin followed by decitraconylation. The five derivatives are readily deacetylated with hydroxylamine to yield the corresponding sulfhydryl insulin derivatives.
Nanami
M
M
Faculty of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
Zaitsu
K
K
Ohkura
Y
Y
D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D007328:Insulin / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000302:isolation & purification; D013864:Thioglycolates / Q000138:chemical synthesis* / Q000302:isolation & purification
D007328:Insulin; D013864:Thioglycolates