可溶性疏水性标签辅助的高效连续流液相多肽合成

doi:10.6023/cjoc202403015

摘要/Abstract

开发绿色、高效的合成工艺是实现治疗性多肽商业化生产的迫切要求. 报道了在微反应器中一种新型疏水性硅基载体双(4-((叔丁基二甲基硅基)氧基)苯基)甲胺(SPPM)辅助进行连续流动液相多肽合成(LPPS)的方法, 该方案由酰胺化模块(微通道反应器, 偶联时间9.0 s)、脱保护模块(填充床反应器, 脱保护时间31.4 s)和萃取洗涤模块(微通道混合器, 洗涤18.0 s)组成. 采用绿色溶剂(用乙酸乙酯代替被限制使用的N,N-二甲基甲酰胺)和N-苄氧羰基氨基酸(用氢解脱保护代替易制毒管制化学品哌啶脱保护), 合成了高纯度的(粗品收率>90%)五肽-3. 这种精确控制反应时间和温度的连续流方法, 从绿色和可持续的角度出发, 为大规模多肽合成开辟了新的前景.

关键词: 连续流动, 微反应器, 可溶性载体, 液相多肽合成

The development of a green and efficient synthetic process is an urgent requirement for the commercial production of therapeutic peptides. In a microreactor, a novel hydrophobic silicon-based carrier bis(4-((tert-butyldimethylsilyl)oxy)- phenyl)methylamine (SPPM) assisted continuous flow liquid phase polypeptide synthesis (LPPS) method was reported. The protocol consists of an amidation module (microchannel reactor, coupling time 9.0 s), a deprotection module (packed bed reactor, deprotection time 31.4 s) and an extraction and washing module (microchannel mixer, washing 18.0 s). High-purity (crude yield >90%) pentapeptide-3 was synthesized by using green solvent (ethyl acetate instead of restricted N,N-dimethyl- formamide) and N-benzyloxycarbonyl-amino acid (hydrogenolysis instead of piperidine deprotection). This continuous flow method with precise control of short reaction time and temperature opens up new prospects for large-scale peptide synthesis from a green and sustainable perspective.

Key words: continuous flow, microreactor, soluble carrier, liquid phase peptide synthesis

引用此文

彭伟, 程蓉, 刘豪, 刘冬梅, 苏贤斌. 可溶性疏水性标签辅助的高效连续流液相多肽合成[J]. 有机化学, 2024, 44(9): 2876-2888.

Wei Peng, Rong Cheng, Hao Liu, Dongmei Liu, Xianbin Su. Highly Efficient Continuous Flow Liquid-Phase Peptide Synthesis Using a Soluble Hydrophobic Tag[J]. Chinese Journal of Organic Chemistry, 2024, 44(9): 2876-2888.

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Entry	Cbz-AA-OH	Solvent	Flow rate/(mL•min^－1)	Reaction time^a/s	Conversion^b/%	Yield^c/%
1	Cbz-Ala-OH	EA	2.0	45.0	>99	96
2	Cbz-Ala-OH	THF	2.0	45.0	27	22
3	Cbz-Ala-OH	DCM	2.0	45.0	87	72

Entry	Cbz-AA-OH	Solvent	Flow rate/(mL•min^－1)	Reaction time^a/s	Conversion^b/%	Yield^c/%
1	Cbz-Ala-OH	EA	2.0	45.0	>99	96
2	Cbz-Ala-OH	THF	2.0	45.0	27	22
3	Cbz-Ala-OH	DCM	2.0	45.0	87	72

Entry	Cbz-AA-OH	Solvent	Flow rate/(mL•min^－1)	Reaction time^a/s	Conversion^b/%	Yield^c/%
1	Cbz-Tyr(^tBu)-OH	EA	2.0	45.0	>99	94
2	Cbz-Ser(^tBu)-OH	EA	2.0	45.0	>99	94
3	Cbz-Lys(Boc)-OH	EA	2.0	45.0	>99	92

Entry	Cbz-AA-OH	Solvent	Flow rate/(mL•min^－1)	Reaction time^a/s	Conversion^b/%	Yield^c/%
1	Cbz-Tyr(^tBu)-OH	EA	2.0	45.0	>99	94
2	Cbz-Ser(^tBu)-OH	EA	2.0	45.0	>99	94
3	Cbz-Lys(Boc)-OH	EA	2.0	45.0	>99	92

Entry	Cbz-AA-OH	Temp./℃	Flow rate/(mL•min^－1)	Reaction time^a/s	Conversion^b/%	Yield^c/%
1	Cbz-Ser(^tBu)-OH	r.t.	1.0	90.0	>99	94
2	Cbz-Ser(^tBu)-OH	r.t.	2.0	45.0	>99	94
3	Cbz-Ser(^tBu)-OH	r.t.	5.0	18.0	>99	92
4	Cbz-Ser(^tBu)-OH	r.t.	10.0	9.0	>99	95
5	Cbz-Ser(^tBu)-OH	r.t.	15.0	6.0	>99	93
6	Cbz-Ser(^tBu)-OH	30	1.0	90.0	>99	92
7	Cbz-Ser(^tBu)-OH	30	2.0	45.0	>99	93
8	Cbz-Ser(^tBu)-OH	30	5.0	18.0	>99	96
9	Cbz-Ser(^tBu)-OH	30	10.0	9.0	>99	98
10	Cbz-Ser(^tBu)-OH	30	15.0	6.0	>99	93
11	Cbz-Ser(^tBu)-OH	40	1.0	90.0	>99	92
12	Cbz-Ser(^tBu)-OH	40	2.0	45.0	>99	94
13	Cbz-Ser(^tBu)-OH	40	5.0	18.0	>99	92
14	Cbz-Ser(^tBu)-OH	40	10.0	9.0	>99	97
15	Cbz-Ser(^tBu)-OH	40	15.0	6.0	>99	93
16	Cbz-Ser(^tBu)-OH	50	1.0	90.0	>99	91
17	Cbz-Ser(^tBu)-OH	50	2.0	45.0	>99	93
18	Cbz-Ser(^tBu)-OH	50	5.0	18.0	>99	92
19	Cbz-Ser(^tBu)-OH	50	10.0	9.0	>99	95
20	Cbz-Ser(^tBu)-OH	50	15.0	6.0	>99	93