微通道连续流动高效绿色合成亮丙瑞林

doi:10.7503/cjcu20200032

摘要/Abstract

摘要：

开发了一种高效绿色的连续流动多肽合成方法, 并成功应用于一种含有9个氨基酸的促性腺激素释放激素类似物(亮丙瑞林)的合成. 该方法采用苄氧羰基(Cbz)保护氨基酸, 在微通道反应器中实现高效偶联与水洗萃取除杂, 并通过钯碳催化剂填充柱氢解反应实现快速洁净地脱除Cbz保护基, 使原料和溶剂的消耗量大大减少, 原子经济性大幅提高. 该高效绿色合成方法将在多肽制药工业中得到更多应用.

关键词: 微通道反应器, 亮丙瑞林, 连续流动, 液相合成

Abstract:

A high efficient and green approach was repored for the synthesis of leuprorelin which is a nonapeptide analogue of gonadotrophin releasing hormone(GnRH) used to treat a wide range of sex hormone-related disorders-based on a continuous-flow system. Benzyloxycarbonyl(Cbz)-protected amino acids were used in this approach and coupling reaction was carried out in micro-channel reactors efficiently followed by extraction to remove impurities. A rapid and clean deprotection of Cbz then successfully achieved through hydrogenolysis via a column packed with Pd/C catalyst. This approach enabled the stiochiometric amidation of peptide chain elongation and catalytic deprotection hence the consumption of raw materials were dramatically reduced. This new strategy will find more application in peptide manufacturing.

Key words: Microchannel reactor, Leuprorelin, Continuous-flow, Liquid phase synthesis

中图分类号:

O629.72

TrendMD:

李士杰,杨洋,崔营营,苏贤斌. 微通道连续流动高效绿色合成亮丙瑞林. 高等学校化学学报, 2020, 41(7): 1559.

LI Shijie,YANG Yang,CUI Yingying,SU Xianbin. High Efficient and Green Approach to the Synthesis of Leuprolide in Continuous-flow Microreactor^†. Chem. J. Chinese Universities, 2020, 41(7): 1559.

图/表 6

参考文献 35

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Compd.	¹H NMR, δ	ESI-MS, m/z
1	7.36—7.31(m, 5H, ArH), 5.21—5.07(m, 2H, CCH₂O), 4.31(d, J=7.0 Hz, 1H, CH), 3.54—	277.14[M+H]⁺
	3.23(m, 4H, NCH₂C), 2.36—1.88(m, 4H, CH₂CH₂), 1.05(t, J=8.4 Hz, 3H, CH₃)
2	4.10(t,J=7.8 Hz, 1H, NCHCO), 3.24—3.10(m, 4H, NCH₂), 2.27—1.80(m, 4H, CH₂CH₂),	143.22[M+H]⁺
	1.05(t, J=7.3 Hz, 3H, CH₃)
3	7.38—7.33(m, 5H, ArH), 5.1(m, 2H, CH₂O), 4.55—4.33(m, 2H, NCH), 3.71—3.24(m,	633.54[M+H]⁺
	6H, NCH₂), 2.37—2.02(m, 4H, CH₂CH₂), 1.53(m, 2H, CH₂), 1.5(s, 18H, CH₃), 1.28—
	1.25(m, 2H, CH₂), 1.09(t, J=7.2 Hz, 3H, CH₃)
17		2021.10[M+Na]⁺
18		1865.09[M+H]⁺
19	8.60—6.61(m, 23H, ArH, NHCO, Indole—H, Imidazole—H), 4.58—3.97(m, 9H, NCHCO),	1209.63[M+H]⁺
	3.66—3.45(m, 2H, OCH₂), 3.18—2.71(m, 12H, NCH₂), 2.28—1.33(m, 18H, CCH₂,
	CCH), 1.03—0.75(m, 15H, CH₃)

Compd.	¹H NMR, δ	ESI-MS, m/z
1	7.36—7.31(m, 5H, ArH), 5.21—5.07(m, 2H, CCH₂O), 4.31(d, J=7.0 Hz, 1H, CH), 3.54—	277.14[M+H]⁺
	3.23(m, 4H, NCH₂C), 2.36—1.88(m, 4H, CH₂CH₂), 1.05(t, J=8.4 Hz, 3H, CH₃)
2	4.10(t,J=7.8 Hz, 1H, NCHCO), 3.24—3.10(m, 4H, NCH₂), 2.27—1.80(m, 4H, CH₂CH₂),	143.22[M+H]⁺
	1.05(t, J=7.3 Hz, 3H, CH₃)
3	7.38—7.33(m, 5H, ArH), 5.1(m, 2H, CH₂O), 4.55—4.33(m, 2H, NCH), 3.71—3.24(m,	633.54[M+H]⁺
	6H, NCH₂), 2.37—2.02(m, 4H, CH₂CH₂), 1.53(m, 2H, CH₂), 1.5(s, 18H, CH₃), 1.28—
	1.25(m, 2H, CH₂), 1.09(t, J=7.2 Hz, 3H, CH₃)
17		2021.10[M+Na]⁺
18		1865.09[M+H]⁺
19	8.60—6.61(m, 23H, ArH, NHCO, Indole—H, Imidazole—H), 4.58—3.97(m, 9H, NCHCO),	1209.63[M+H]⁺
	3.66—3.45(m, 2H, OCH₂), 3.18—2.71(m, 12H, NCH₂), 2.28—1.33(m, 18H, CCH₂,
	CCH), 1.03—0.75(m, 15H, CH₃)

Entry	Amino acid	Efficiency of one wash in a reaction caldron(%)	Efficiency of one wash in a microchannel reactor(%)
1	Z-Pro-OH	100	100
2	Z-Arg(Boc)₂-OH	32.5	56.4
3	H-Arg(Boc)₂-OH	100	100
4	Z-Leu-OH	100	100
5	Z-D-Leu-OH	100	100
6	Z-Tyr(tBu)-OH	35	52.2
7	H-Tyr(tBu)-OH	100	100
8	Z-Ser(tBu)-OH	100	100
9	Z-Trp(Boc)-OH	20.2	31.6
10	H-Trp(Boc)-OH	100	100
11	Z-His(Trt)-OH	8.2	15.4
12	H-His(Trt)-OH	100	100
13	Z-Pyr-OH	100	100

Entry	Amino acid	Efficiency of one wash in a reaction caldron(%)	Efficiency of one wash in a microchannel reactor(%)
1	Z-Pro-OH	100	100
2	Z-Arg(Boc)₂-OH	32.5	56.4
3	H-Arg(Boc)₂-OH	100	100
4	Z-Leu-OH	100	100
5	Z-D-Leu-OH	100	100
6	Z-Tyr(tBu)-OH	35	52.2
7	H-Tyr(tBu)-OH	100	100
8	Z-Ser(tBu)-OH	100	100
9	Z-Trp(Boc)-OH	20.2	31.6
10	H-Trp(Boc)-OH	100	100
11	Z-His(Trt)-OH	8.2	15.4
12	H-His(Trt)-OH	100	100
13	Z-Pyr-OH	100	100

Step	Peptide	First-round efficiency(%)	Second-round efficiency(%)	Third-round efficiency(%)
1	Z-Pro-NHEt	100	—	—
2	Z-Arg(Boc)₂-Pro-NHEt	97.3	100	—
3	Z-Leu-Arg(Boc)₂-Pro-NHEt	91.4	100	—
4	Z-D-Leu-Leu-Arg(Boc)₂-Pro-NHEt	89.7	100	—
5	Z-Tyr(tBu)-D-Leu-Leu-Arg(Boc)₂-Pro-NHEt	94.1	100	—
6	Z-Ser(tBu)-Tyr(tBu)-D-Leu-Leu-Arg(Boc)₂-Pro-NHEt	92.1	100	—
7	Z-Trp(Boc)-Ser(tBu)-Tyr(tBu)-D-Leu-Leu-Arg(Boc)₂-Pro-NHEt	82.6	100	—
8	Z-His(Trt)-Trp(Boc)-Ser(tBu)-Tyr(tBu)-D-Leu-Leu-Arg(Boc)₂-Pro-NHEt	33.5	76.2	100
9	Z-Pyr-His(Trt)-Trp(Boc)-Ser(tBu)-Tyr(tBu)-D-Leu-Leu-Arg(Boc)₂-Pro-NHEt	79.6	100	—