A novel method for repetitive peptide synthesis in solution without isolation of intermediates
A novel method for repetitive peptide synthesis in solution without isolation of intermediates
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DOI：
10.1002/psc.670
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发表时间：
2005-10-01
影响因子：
2.1
通讯作者：
Ter Voert, EJM
中科院分区：
文献类型：
--
作者：
Eggen, IF;Bakelaar, FT;Ter Voert, EJM
开发了一种新的溶液中大规模生产多肽的方法，称为DioRaSSP-DiSynth多肽快速溶液合成法。该方法结合了经典溶液相合成法的均相特性和固相法固有的通用性和易于自动化的优点。该过程包括在一个永久性有机相中的偶联和去保护的重复循环，并且进一步的特点是中间体不是孤立的。偶联是由水溶性碳二亚胺介导的。根据实际多肽的序列，可以应用几种类型的功能来进行暂时的氨基保护，包括Z、Fmoc、MSC和NSC。在Z基团氢解过程中，For-mate是首选的供氢体，而1，8-二氮杂双环[5.4.0]Undec-7-ene用于脱保护Fmoc、MSC和NSC。在MSC和NSC脱保护过程中加入吗啉以清除产生的烯烃。根据这种高效的合成方法，工艺容易放大，并产生可重复使用的高纯度产品，这是由一种新的残留活性化合物的猝灭方法保证的，该方法应用了形成阴离子的胺，如β-丙氨酸酯。这种酯应该表现出类似于临时氨基保护功能的不稳定性，允许同时去保护生长肽和猝灭的化合物。DioRaSSP方法确保在下一个合成循环的耦合步骤之前，通过碱性水萃取(即活性)完全定量去除脱保护的猝灭化合物，而由于存在疏水保护功能，生长肽仍固定在有机相中。版权所有(C)2005欧洲肽协会和John Wiley&Sons，Ltd.
A novel method was developed for the large-scale manufacture of peptides in solution, called DioRaSSP - Diosynth Rapid Solution Synthesis of Peptides. This method combines the advantages of the homogeneous character of classical solution-phase synthesis with the universal character and the amenability to automation inherent to the solid-phase approach. The process consists of repetitive cycles of coupling and deprotection in a permanent organic phase and is further characterized by the fact that intermediates are not isolated. Couplings are mediated by water-soluble carbodiimide. Several types of function may be applied for temporary amino protection depending on the sequence of the actual peptide, including Z, Fmoc, Msc and Nsc. For-mate is the prefer-red hydrogen donor during hydrogenolysis of the Z function, while 1,8-diazabicyclo[5.4.0]undec-7-ene is used to deprotect Fmoc, Msc and Nsc. Morpholine is added during the deprotection of Msc and Nsc to scavenge the arising alkenes. Processes according to this highly efficient synthesis method are easy to scale up and yield products of reproducible high purity, which is guaranteed by a new quenching method for residual activated compounds, applying an anion-forming amine such as a beta-alanine ester. This ester should display a lability similar to that of the temporary amino-protecting function, allowing simultaneous deprotection of the growing peptide and the quenched compound. The DioRaSSP approach assures the completely quantitative removal of deprotected quenched compounds before the coupling step of the next cycle of the synthesis by basic aqueous (that is active) extraction, while the growing peptide remains anchored in the organic phase due to the presence of hydrophobic protecting functions. Copyright (c) 2005 European Peptide Society and John Wiley & Sons, Ltd.