PROCESS FOR THE MANUFACTURE OF HUMAN INSULIN

摘要

1521919 Human insulin HOECHST AG 15 Dec 1975 [21 Dec 1974] 51191/75 Heading C3H A process for the manufacture of human insulin, which comprises condensing des-B23-30- octapeptide porcine insulin of the general Formula I wherein R represents a protective amino group eliminable by proton solvolysis ot #-elimination with a peptide of Formula II H-Gly-Phe-Phe- Tyr(X) - Thr(X) - Pro - Lys(Boc) - Thr(But) - OBut wherein X is H or t-butyl, and eliminating the protective groups by proton solvolysis or #-elimination. R is preferably t-butyloxycarbonyl, t-amyloxycarbonyl or methylsulphonylethyloxycarbonyl and the protective groups may be removed by trifluoroacetic acid or an alkali. When R is Boc, equimolar amounts of I and II may be reacted with 1-hydroxybenzotriazole as condensation agent in the presence of a less than equimolar amount of dicyclohexylcarbodiimide. Prior to removal of protective groups, the human insulin may be purified by partition chromatography, preferably in n-butanal/glacial acetic acid/water (2 : 1 : 10 by volume) or gel chromatography. Peptide intermediates isolated are:- N, N-bis-Boc-des-B23-30-octapeptide porcine insulin. Octapeptides: Z and H-Gly-Phe-Phe-Tyr- (But) -Thr-Pro-Lys(Boc) -Thr(But) -OBut. Pentapeptides: Z-Gly-Phe-Phe-Tyr(But)-Thr- OCH3 and OH Tripeptides: Z-Gly-Phe-Phe-OBut and OH; H and Z-Pro-Lys(Boc)-Thr(But)-OBut. Dipeptide intermediates isolated are: Z and H-Phe-Phe-OBut; Z and H-Tyr(But)-Thr-OCH3; Z and H-Lys(Boc)-Thr(But)-OBut.