- Piperazine and DBU: A safer alternative for rapid and efficient Fmoc deprotection in solid phase peptide synthesis
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In Solid Phase Peptide Synthesis (SPPS), contamination with deletion sequences which often co-elute with the target peptide continues to be a major challenge as these impurities can significantly affect the target peptide's properties. Here, we report an efficient Fmoc-deprotection solution containing piperazine and DBU which can cause complete removal of the Fmoc group in less than a minute. This combination rivals piperidine in speediness as revealed by kinetic studies. We demonstrate the efficiency of the piperazine/DBU solution by synthesizing the polyAla stretch with a significant reduction of deletion products occurring due to partial Fmoc deprotection. We verify the utility of the deprotection solution by successfully synthesizing four aggregation prone difficult peptide sequences. We further demonstrate that this combination can also be used to synthesize aspartimide and epimerization prone sequences when supplemented with 1% formic acid and is compatible with 2-chlorotrityl chloride resin. We conclude that piperazine/DBU can be used as a safer and effective alternative to piperidine in Fmoc-SPPS.
- Ralhan, Krittika,KrishnaKumar, V. Guru,Gupta, Sharad
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p. 104417 - 104425
(2015)
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- Evaluation of acid-labile S-protecting groups to prevent Cys racemization in Fmoc solid-phase peptide synthesis
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Phosphonium and uronium salt-based reagents enable efficient and effective coupling reactions and are indispensable in peptide chemistry, especially in machine-assisted SPPS. However, after the activating and coupling steps with these reagents in the presence of tertiary amines, Fmoc derivatives of Cys are known to be considerably racemized during their incorporation. To avoid this side reaction, a coupling method mediated by phosphonium/uronium reagents with a weaker base, such as 2,4,6-trimethylpyridine, than the ordinarily used DIEA or that by carbodiimide has been recommended. However, these methods are appreciably inferior to the standard protocol applied for SPPS, that is, a 1 min preactivation procedure of coupling with phosphonium or uronium reagents/DIEA in DMF, in terms of coupling efficiency, and also the former method cannot reduce racemization of Cys(Trt) to an acceptable level (a high temperature.
- Hibino, Hajime,Miki, Yasuyoshi,Nishiuchi, Yuji
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