Madder Annemieke, Li Liu, De Muynck Hilde, Farcy Nadia, Van Haver Dirk, Fant Franky, Vanhoenacker Gerd, Sandra Pat, Davis Anthony P, De Clercq Pierre J
Laboratory of Organic Synthesis, Department of Organic Chemistry, Ghent University, Krijgslaan 281, B-9000 Gent, Belgium.
J Comb Chem. 2002 Nov-Dec;4(6):552-62. doi: 10.1021/cc020016g.
A series of peptidosteroid derivatives containing two independent peptide chains in which Ser and His are incorporated were synthesized by solid-phase peptide synthesis. The activity of the different compounds in the hydrolysis of the activated substrate NF31 was assessed in a stepwise fashion. First, the different resin-bound derivatives 6a-l and 6x-z were individually assayed for serine esterification in the absence of water. The use of a colored substrate allowed for a visual identification of the most active compounds. Through the inclusion of control substances, the involvement of histidine in the mechanism for serine acylation was shown. Second, the hydrolysis and methanolysis of the different acylated derivatives 8a-l and 8x were evaluated using UV spectroscopy, again indicating the involvement of histidine. The feasibility of applying the above procedures in a combinatorial context was proven via the screening of artificial libraries, created by mixing the different resin-bound peptidosteroid compounds. In this respect, the use of a photocleavable linker allowed for the unambiguous structural characterization of the selected members via application of single-bead electrospray tandem mass spectrometry.
通过固相肽合成法合成了一系列含有两条独立肽链且引入了丝氨酸（Ser）和组氨酸（His）的肽甾体衍生物。以逐步方式评估了不同化合物对活化底物NF31的水解活性。首先，在无水条件下分别测定不同的树脂结合衍生物6a-l和6x-z的丝氨酸酯化反应。使用有色底物可直观鉴定出活性最高的化合物。通过加入对照物质，证明了组氨酸参与丝氨酸酰化机制。其次，使用紫外光谱法评估了不同酰化衍生物8a-l和8x的水解和甲醇解反应，再次表明组氨酸的参与。通过筛选由不同树脂结合的肽甾体化合物混合创建的人工文库，证明了上述程序在组合环境中应用的可行性。在这方面，使用光可裂解连接子可通过单珠电喷雾串联质谱法对所选成员进行明确的结构表征。
