Studies on the Sulfated Tyrosine Containing Peptides Using a Novel Solid-Phase Synthetic Method

新型固相合成方法研究硫酸化酪氨酸肽

基本信息

批准号：
10672008
负责人：
KITAGAWA Kouki
金额：
$ 2.37万
依托单位：
Niigata College of Pharmacy
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2000
项目状态：
已结题

项目摘要

1. Based on the kinetic studies concerning the desulfation of Tyr (SO_3Na) and the deprotection of the protecting groups, Arg (Pbf) and Ser (^tBu), in TFA, we proposed 90% aqueous TFA treatment at low temperature is a suitable deprotection/cleavage protocol for the direct synthesis of Tyr (SO_3H)-containing peptides.2. An efficient Fmoc-based solid-phase method for the synthesis of Tyr (SO_3H)-containing peptides was developed. This approach involves two key features : (1) use of the 2-chlorotrityl resin as a solid support, and (2) a S_N1-type deprotection/cleavage protocol based on the TFA-mediated acidolysis at low temperature. Various molecular forms of gastrin-II and cholecystokinin (CCK) involving big gastrin-II and CCK-39 were prepared by this approach without notable difficulty.3. Application of the Fmoc-based solid-phase segment condensation approach to the synthesis of human big gastrin-II and its C-terminal Gly-extended form (G34-Gly sulfate) was investigated. In these synthe … More ses, 2-chlorotrityl resin was exclusively employed for an anchor resin to prepare the three peptide segments having the C-terminal Pro residue and the Tyr (SO_3H)-containing resin-bound segment.4. Based on the mass spectrometric behaviors of the various Tyr (SO_3H)-containing peptides, we indicated that an anionic Tyr (SO_3H) residue tends to form a stable conjugate acid-base pair with cationin functional groups, especially the guanidine function of the Arg residues, in aqueous solutions and under nonpolar conditions.5. The big-molecular-form cholecystokinin (CCK)-peptides, CCK-39 and CCK-58, were prepared by the thioester segment condensation method using two or three peptide segments (the C-terminal Tyr (SO_3H)-containing segment and the partially protected thioester segments having C-terminal Pro residues). A brief TFA treatment of the final condensation product afforded objective CCK-39 and CCK-58, respectively6. Deprotection methods of the protecting groups for Cys and disulfide bond formation methods, in which the desulfation from the Tyr (SO_3H) residue was not associated, were examined using oxytocin sulfate as a model peptide. Corn-snail toxin, a-conotoxin Epl, was synthesized using Cys (Trt) protecting group and TFA-mediated acidolysis at low temperature followed by the air oxidation to form two disulfide bonds. Less

1.基于TFA中Tyr（SO_3Na）脱硫和保护基Arg（Pbf）和Ser（^tBu）脱保护的动力学研究，我们提出低温下90％TFA水溶液处理是合适的直接合成含Tyr (SO_3H)肽的脱保护/裂解方案。2.一种基于Fmoc的高效固相合成Tyr方法。开发了含 (SO_3H) 的肽，该方法涉及两个关键特征：(1) 使用 2-氯三苯甲基树脂作为固体支持物，以及 (2) 基于 TFA 介导的酸解的 S_N1 型脱保护/裂解方案。通过这种方法在低温下制备了各种分子形式的胃泌素-II 和缩胆囊素 (CCK)，其中包括大胃泌素-II 和 CCK-39。困难.3. 研究了基于 Fmoc 的固相片段缩合方法在人大胃泌素-II 及其 C 末端甘氨酸延伸形式（G34-甘氨酸硫酸盐）的合成中的应用。 2-氯三苯甲基树脂专门用于锚定树脂，以制备具有C端Pro残基和含有Tyr（SO_3H）的树脂结合的三个肽段4.基于各种含Tyr (SO_3H)的肽的质谱行为，我们表明阴离子Tyr (SO_3H)残基倾向于与阳离子官能团，尤其是胍官能团形成稳定的共轭酸碱对。水溶液中和非极性条件下的 Arg 残基。5。 CCK-39和CCK-58是使用两个或三个肽片段（C端含有Tyr（SO_3H）的片段和具有C端Pro残基的部分受保护的硫酯片段）通过硫酯片段缩合方法制备的。最终缩合产物的 TFA 处理分别提供了针对 Cys 和二硫键形成的保护基团的目标 CCK-39 和 CCK-58 脱保护方法6。使用硫酸催产素作为模型肽，使用 Cys (Trt) 保护基和 TFA 介导合成玉米蜗牛毒素 a-芋螺毒素 Epl，对其中 Tyr (SO_3H) 残基的脱硫不相关的方法进行了检查。低温酸解，随后空气氧化，形成两个二硫键。