Highly Efficient Continuous Flow Liquid-Phase Peptide Synthesis Using a Soluble Hydrophobic Tag

doi:10.6023/cjoc202403015

Abstract

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

Cite this article

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.

Export EndNote|Reference Manager|ProCite|BibTeX|RefWorks

share this article

Fig. & Tab. 16

References 41

[1]	(a) Lau, J. L.; Dunn, M. K. Bioorg. Med. Chem. Lett. 2018, 26, 2700.
	(b) Tanemura, Y.; Mochizuki, Y.; Kumachi, S. Biology 2015, 4, 161.
	(c) Lin, K. J. V.; Emrick, J. J.; Kelly, M. J. S.; Herzig, V. Cell 2019, 178, 1362.
	(d) Drucker, D. J. Nat. Rev. Drug Discovery 2020, 19, 277.
[2]	(a) Thomas, K.; Markus, M. Methods Mol. Bol. 2022, 2384, 175.
	(b) Ceccato, M. L.; Chenu, J.; Mery, J.; Follet, M.; Calas, B. Tetrahedron Lett. 1990, 31, 6189.
	(c) Nicolas, E.; Clemente, J.; Ferrer, T. Tetrahedron 1997, 53, 3179.
[3]	(a) Vincent, d. V.; Charlotte, R. J. Am. Chem. Soc. 1953, 75, 4879.
	(b) Fan, J. H.; Bian, Y. N.; Su, X. B. Chem. J. Chin. Univ. 2018, 39, 2679 (in Chinese).
	(范佳辉, 卞亚楠, 苏贤斌, 高等学校化学学报, 2018, 39, 2679.) doi: 10.7503/cjcu20180350
	(c) Lawrenson, S. B.; Arav, R.; North, M. Green Chem. 2017, 19, 1685.
[4]	Rasmussen, J. H. Bioorg. Med. Chem. 2018, 26, 2914.
[5]	(a) Jadhav, S.; Seufert, W.; Lechner, C. Chimia 2021, 75, 476.
	(b) Rasmussen, J. H. Bioorg. Med. Chem. 2018, 26, 2914.
[6]	Eggen, I. F.; Bakelaar, F. T.; Petersen, A. J. Pept. Sci. 2005, 11, 633. pmid: 15880381
[7]	Merrifield, R. B. J. Am. Chem. Soc. 1963, 85, 2149.
[8]	Yin, H.; Chen, X. T. Acta Chim. Sinica 2022, 80, 444 (in Chinese).
	(尹昊, 陈西同, 化学学报, 2022, 80, 444.) doi: 10.6023/A21120580
[9]	Zhuang, H.; Hao, H.; Qiu, Y. Chin. J. Chem. 2023, 41, 2010.
[10]	Li, T.; Zhang, Y. Peng, P. Chin. J. Chem. 2022, 40, 1571.
[11]	Al Musaimi, O.; de la Torre, B. G.; Albericio, F. Green Chem. 2020, 22, 996.
[12]	(a) Lawrenson, S. B.; Arav, R.; North, M. Green Chem. 2017, 19, 1685.
	(b) Bonnamour, J.; Metro, T. X.; Martinez, J.; Lamaty, F. Green Chem. 2013, 15, 1116.
[13]	(a) Egelund, P. H. G.; Jadhav, S. ACS Sustainable Chem. Eng. 2021, 9, 14202.
	(b) Martin, V.; Jadhav, S.; Egelund, P. H. G. Green Chem. 2021, 23, 3295.
	(c) Xu, B. F.; Yang, S.; Zhu, J. M.; Ma, Y. D.; Zhao, G.; Guo, Y.; Xu, L. Chem. Res. Chin. Univ. 2014, 30, 103.
[14]	Shemyakin, M. M.; Ovchinnikov, Y. A. Tetrahedron Lett. 1965, 6, 2323.
[15]	Pillai, V. N. R.; Mutter, M.; Bayer, E. J. Org. Chem. 1980, 45, 5364.
[16]	(a) Bayer, E.; Mutter, M. Nature 1972, 237, 512. pmid: 4427055
	(b) Bayer, E.; Mutter, M.; Uhmann, R.; Polster, J.; Mauser, H. J. Am. Chem. Soc. 1974, 96, 7333. pmid: 4427055
	(c) Musaimi, O.; Albericio, F. Green Chem. 2020, 22, 996. pmid: 4427055
	(d) Mutter, M.; Hagenmaier, H.; Bayer, E. Angew. Chem. Int. Ed. 1971, 10, 811. pmid: 4427055
	(e) Mutter, M.; Bayer, E. Angew. Chem. Int. Ed. 1974, 13, 88. pmid: 4427055
	(f) Bayer, E.; Mutter, M. Chem. Ber. 1974, 107, 1344. pmid: 4427055
[17]	Hitoshi, T.; Tomoyuki, O. Bull. Chem. Soc. Jpn. 2001, 74, 733.
[18]	Takahashi, D.; Yamamoto, T. Tetrahedron Lett. 2012, 53, 1936.
[19]	Takahashi, D. Org. Lett. 2012, 14, 4514. doi: 10.1021/ol302002g pmid: 22920411
[20]	(a) Li, H.; Chao, J.; Qin, C. G. J. Org. Chem. 2020, 85, 6271.
	(b) Li, H.; Chao, J.; Qin, C. G. Org. Chem. Front. 2020, 7, 689.
	(c) Li, H.; Chao, J.; Qin, C. G. Org. Lett. 2020, 22, 3323.
[21]	Lu, D.; Wang, S.; Yin, H. Synlett 2020, 31, 1163.
[22]	(a) Tetsu, T.; Hidekazu, O.; Shu, K. Nature 2015, 520, 329.
	(b) Sheng, B. S.; Li, C.; Liu, Y. Y.; Wang, A. J.; Wang, Y.; Zhang, J.; Liu, W. X. Chem. J. Chin. Univ. 2019, 40, 1365.
[23]	Adamo, A.; Beingessner, R. L.; Behnam, M. Science 2016, 352, 61.
[24]	Szloszar, A. ChemistrySelect 2017, 2, 6036.
[25]	Audubert, C.; Gamboa, M. O. J.; Lebel, H. Angew. Chem. 2017, 129, 6391.
[26]	(a) Cheng, D.; Chen, F. E. Chem. Ind. Eng. Prog. 2019, 38, 556 (in Chinese).
	(程荡, 陈芬儿, 化工进展, 2019, 38, 556.) doi: 10.16085/j.issn.1000-6613.2018-1174
	(b) Gordon, C. P. Org. Biomol. Chem. 2018, 16, 180.
[27]	Loubiere, K.; Oelgemoller, M.; Aillet, T.; Dechy-Cabaret, O.; Prat, L. Chem. Eng. Process. 2016, 104, 120.
[28]	Mizuno, K.; Nishiyama, Y.; Ogaki, T.; Terao, K.; Ikeda, H.; Kakiuchi, K. J. Photochem. Photobiol., C 2016, 29, 107.
[29]	Fuse, S.; Otake, Y.; Nakamura, H. Eur. J. Org. Chem. 2017, 6466.
[30]	Otake, Y.; Nakamura, H.; Fuse, S. Tetrahedron Lett. 2018, 59, 1691.
[31]	Politano, F.; Oksdath-Mansilla, G. Org. Process Res. Dev. 2018, 22, 1045.
[32]	Di Filippo, M.; Bracken, C.; Baumann, M. Molecules 2020, 25, 356.
[33]	Bacsa, B.; Kappe, C. O. Nat. Protoc. 2007, 2, 2222.
[34]	Szloszar, A.; Mandity, I. M. J. Flow Chem. 2018, 8, 21.
[35]	Hartrampf, N.; Saebi, A.; Poskus, M.; Gates, Z. P.; Pentelute, B. L. Science 2020, 368, 980. doi: 10.1126/science.abb2491 pmid: 32467387
[36]	Mandity, I. M.; Olasz, B.; Otvos, S. B.; Fulop, F. ChemSusChem 2014, 7, 3172.
[37]	Nishimura, S.; Higashi, N.; Koga, T. J. Polym. Sci., Part A: Polym. Chem. 2019, 10, 71.
[38]	Fuse, S.; Mifune, Y.; Takahashi, T. Angew. Chem. Int. Ed. 2014, 53, 851.
[39]	Otake, Y. Adachi, K. Fuse, S. React. Chem. Eng. 2023, 8, 863.
[40]	Li, S. J.; Yang, Y.; Cui, Y. Y.; Su, X. B. Chem. J. Chin. Univ. 2020, 41, 1559 (in Chinese).
	(李士杰, 杨洋, 崔营营, 苏贤斌, 高等学校化学学报, 2020, 41, 1559.) doi: 10.7503/cjcu20200032
[41]	Sabatini, M. T.; Boulton, L. T.; Sneddon, H. F. Nat. Catal. 2019, 2, 10. doi: 10.1038/s41929-018-0211-5

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	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

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

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

可溶性疏水性标签辅助的高效连续流液相多肽合成