標題: Toward sustainable solid-phase peptide synthesis strategy – in situ Fmoc removal

發(fā)表時間: 2024

發(fā)表作者: Ndumiso Mthethwa, K. P. Nandhini, Ashish Kumar, Anamika Sharma, Beatriz G. de la Torre, Fernando Albericio

發(fā)表機構(gòu):

• Peptide Science Laboratory, School of Chemistry and Physics, University of KwaZulu-Natal, Durban, South Africa
• KwaZulu-Natal Research Innovation and Sequencing Platform (KRISP), School of Laboratory Medicine and Medical Sciences, College of Health Sciences, University of KwaZulu-Natal, Durban, South Africa
• Department of Organic Chemistry, University of Barcelona, Barcelona, Spain

關(guān)鍵詞: green chemistry, in situ coupling deprotection, peptide, 4-methylpiperidine, sustainable chemistry

ABSTRACT
Solid-phase peptide synthesis (SPPS) is the method of choice for the synthesis of peptides for research and production purposes. Despite having several positive features, it remains a challenge to reduce the amount of solvent waste generated during the synthesis. We proposed a 3-step protocol (in-situ Fmoc removal) where washing after coupling was eliminated. Here, the in-situ Fmoc removal protocol was optimized by adding an extra 4-methylpiperidine (4-MP) treatment to ensure the removal of the Fmoc. Additionally, a second addition of the  carbodiimide during the coupling step is performed to form an in situ more active ester of the Fmoc-amino acid. The number of washings has been kept to a minimum of three adding in two of them 1% of OxymaPure to ensure the total removal of the 4-MP. This strategy, which saves up to 60% of solvent, was successfully demonstrated in two important Active Pharmaceutical Ingredients (APIs), angiotensin II and afamelanotide synthesis, with high purity. This strategy will be added to the green toolbox of SPPS making the approach more sustainable.


摘要：
固相肽合成（SPPS）是合成研究和生產(chǎn)用肽的首選方法。盡管具有幾個積極特征，但在合成過程中減少溶劑廢物的生成仍然是一個挑戰(zhàn)。我們提出了一種3步協(xié)議（原位Fmoc去除），其中消除了偶聯(lián)后的洗滌步驟。在這里，通過添加額外的4-甲基哌啶（4-MP）處理來優(yōu)化原位Fmoc去除協(xié)議，以確保去除Fmoc。此外，在偶聯(lián)步驟中進行第二次加入碳二亞胺以形成原位更活躍的Fmoc-氨基酸酯。洗滌次數(shù)最少為三次，其中兩次添加1%的OxymaPure以確保完全去除4-MP。這種策略可以節(jié)省高達60%的溶劑，并已成功應(yīng)用于兩種重要的活性藥物成分（APIs），即血管緊張素II和阿法諾肽的合成，并且純度很高。這一策略將被添加到SPPS的綠色工具箱中，使該方法更加可持續(xù)。