Fmoc-Lys(Pac)-OH和青霉素G酰化酶在新型半合成胰岛素类似物制备中的应用。

The use of Fmoc-Lys(Pac)-OH and penicillin G acylase in the preparation of novel semisynthetic insulin analogs.

作者信息

Záková Lenka, Zyka Daniel, Jezek Jan, Hanclová Ivona, Sanda Miloslav, Brzozowski Andrzej M, Jirácek Jirí

机构信息

Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Flemingovo nám. 2, 166 10 Praha 6, Czech Republic.

出版信息

J Pept Sci. 2007 May;13(5):334-41. doi: 10.1002/psc.847.

DOI:10.1002/psc.847

PMID:17436342

Abstract

In this paper, we present the detailed synthetic protocol and characterization of Fmoc-Lys(Pac)-OH, its use for the preparation of octapeptides H-Gly-Phe-Tyr-N-MePhe-Thr-Lys(Pac)-Pro-Thr-OH and H-Gly-Phe-Phe-His-Thr-Pro-Lys(Pac)-Thr-OH by solid-phase synthesis, trypsin-catalyzed condensation of these octapeptides with desoctapeptide(B23-B30)-insulin, and penicillin G acylase catalyzed cleavage of phenylacetyl (Pac) group from Nepsilon-amino group of lysine to give novel insulin analogs [TyrB25, N-MePheB26,LysB28,ProB29]-insulin and [HisB26]-insulin. These new analogs display 4 and 78% binding affinity respectively to insulin receptor in rat adipose membranes.

摘要

在本文中，我们展示了芴甲氧羰基-赖氨酸（2,2,2-三氯苯甲酰基）-OH（Fmoc-Lys(Pac)-OH）的详细合成方案及表征，其通过固相合成用于制备八肽H-甘氨酸-苯丙氨酸-酪氨酸-N-甲基苯丙氨酸-苏氨酸-赖氨酸（2,2,2-三氯苯甲酰基）-脯氨酸-苏氨酸-OH（H-Gly-Phe-Tyr-N-MePhe-Thr-Lys(Pac)-Pro-Thr-OH）和H-甘氨酸-苯丙氨酸-苯丙氨酸-组氨酸-苏氨酸-脯氨酸-赖氨酸（2,2,2-三氯苯甲酰基）-苏氨酸-OH（H-Gly-Phe-Phe-His-Thr-Pro-Lys(Pac)-Thr-OH），这些八肽与去八肽（B23 - B30）-胰岛素经胰蛋白酶催化缩合，以及青霉素G酰化酶催化从赖氨酸的Nε-氨基上裂解掉苯乙酰（Pac）基团以得到新型胰岛素类似物[TyrB25, N-MePheB26,LysB28,ProB29]-胰岛素和[HisB26]-胰岛素。这些新的类似物分别对大鼠脂肪细胞膜中的胰岛素受体显示出4%和78%的结合亲和力。