High Efficient and Green Approach to the Synthesis of Leuprolide in Continuous-flow Microreactor†

doi:10.7503/cjcu20200032

Abstract

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

CLC Number:

O629.72

TrendMD:

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

Figures/Tables 6

References 35

[1]	Hwang T. L., Ranganathan K., Fang Y. Q., Crockett R. D., Osgood S., Cui S., Org. Process. Res. Dev., 2018, 22(8), 1007—1014
[2]	Feng H. Y., Gao L., Ye X. H., Wang L., Xue Z. H., Kong J. M., Li L. Z., Chem. Res. Chinese Universities, 2017, 33(2), 155—159
[3]	Isidro-Llobet A., Kenworthy M. N., Mukherjee S., Kopach M. E., Wegner K., Gallou F., Smith A. G., Roschangar F., J. Org. Chem., 2019, 84(8), 4615—4628
[4]	Lawrenson S. B., Arav R., North M., Green Chem., 2017, 19(7), 1685—1691
[5]	Bonnamour J., Métro T. X., Martinez J., Lamaty F., Green Chem., 2013, 15(5), 1116—1120
[6]	Fan J. H., Bian Y. N., Su X. B., Chem. J. Chinese Universities, 2018, 39(12), 2679—2685
	( 范佳辉, 卞亚楠, 苏贤斌. 高等学校化学学报, 2018, 39(12), 2679—2685)
[7]	Xu B. F., Yang S., Zhu J. M., Ma Y. D., Zhao G., Guo Y., Xu L., Chem. Res. Chinese Universities, 2014, 30(1), 103—107
[8]	Sakakibara S., Biopolymers, 1999, 51(4), 279—296
[9]	Gravert D. J., Janda K. D., Chem. Rev., 1997, 97(2), 489—510 URL pmid: 11848880
[10]	Aihara K., Komiya C., Shigenaga A., Inokuma T., Takahashi D., Otaka A., Org. Lett., 2015, 17(3), 696—699
[11]	Sheng B. S., Li C., Liu Y. Y., Wang. A. J., Wang Y., Zhang J., Liu W. X., Chem. J. Chinese Universities, 2019, 40(7), 1365—137
	( 盛炳琛, 李从, 刘颖雅, 王安杰, 王瑶, 张箭, 刘伟旭. 高等学校化学学报, 2019, 40(7), 1365—1373)
[12]	Tetsu T., Hidekazu O., Shū K., Nature, 2015, 520(7547), 329—332 URL pmid: 25877201
[13]	Adamo A., Beingessner R. L., Behnam M., Chen J., Jamison T. F., Jensen K. F., Monbaliu J. C. M., Myerson A. S., Revalor E. M., Snead D. R., Stelzer T., Weeranoppanant N., Wong S. Y., Zhang P., Science, 2016, 352(6281), 61—67
[14]	Audubert C., Gamboa M. O. J., Lebel H., Angew. Chem., 2017, 129(22), 6391—6394
[15]	Cheng D., Chen F. E., ., Chemical Industry and Engineering Progress, 2019, 38(01), 556—574
	( 程荡, 陈芬儿. 化工进展, 2019, 38(01), 556—574)
[16]	Gordon C. P., Org. Biomol. Chem., 2018, 16(2), 180—196 URL pmid: 29255827
[17]	Fuse S., Otake Y., Nakamura H., Chem. Asian J., 2018, 13(24), 3818—3832 URL pmid: 30341812
[18]	Szloszár A., Fülöp F., Mándity I. M., Chemistry Select, 2017, 2(21), 6036—6039
[19]	Mándity I. M., Olasz B., Ötvös S. B., Fülöp F., ChemSusChem, 2014, 7(11), 3172—3176 URL pmid: 25196512
[20]	Spare L. K., Laude V., Harman D. G., Aldrich-Wright J. R., Gordon C. P., React. Chem. Eng., 2018, 3(6), 875—882
[21]	Mijalis A. J., Thomas D. A., Simon M. D., Adamo A., Beaumont R., Jensen K. F., Pentelute B. L., Nat. Chem. Biol., 2017, 13(5), 464—466 URL pmid: 28244989
[22]	Nishimura S., Higashi N., Koga T., J. Polym. Sci. Pol. Chem. Part A: Polym. Chem., 2019, 10(1), 71—76
[23]	Knudsen R. K., Ladlow M., Bandpey Z., Ley S. V., J. Flow Chem., 2014, 4(1), 18—21
[24]	Mifune Y., Nakamura H., Fuse S., Org. Biomol. Chem., 2016, 14(47), 11244—11249 URL pmid: 27849093
[25]	Fuse S., Mifune Y., Takahashi T., Angew. Chem., 2014, 53(3), 851—855
[26]	Elias Y., von Rohr P. R., Bonrath W., Medlock J., Buss A., Chem. Eng. Process, 2015, 95, 175—185
[27]	Avril A., Hornung C. H., Urban A., Fraser D., Horne M., Veder J. P., Tsanaktsidis J., Rodopoulos T., Henry C., Gunasegaram D. R., React. Chem. Eng., 2017, 2(2), 180—188 doi: 10.1039/C6RE00188B URL
[28]	Takashi O., Claudio B., Joel M. H., Steven V. L., Org. Process Res. Dev., 2014, 18(11), 1560—1566 doi: 10.1021/op500208j URL
[29]	Cossar P. J., Hizartzidis L., Simone M. I., McCluskey A., Gordon C. P., Org. Biomol. Chem., 2015, 13(26), 7119—7130 URL pmid: 26073166
[30]	Han W., Youn H. J., Asian Pac. J. Cancer P., 2019, 20(5), 1475—1479
[31]	Fratta P., Nirmalananthan N., Masset L., Skorupinska I., Collins T., Cortese A., Pemble S., Malaspina A., Fisher E. M. C., Greensmith L., Hanna M. G., Eur. Neurol., 2014, 82(23), 2077—2084
[32]	Masahiko F., Nonapeptide Amide Analogs of Luteinizing Releasing Hormone, US 4008209(A), 1977-02-15
[33]	Nicolas E., Clemente J., Ferrer T., Albericio F., Giralt E., Tetrahedron, 1997, 53(9), 3179—3194 doi: 10.1016/S0040-4020(97)00029-X URL
[34]	Zhang J., Cen T., Wang A. M., Zhou C., Du Z. Q., Wu X. M., Shen S. B., Chemical Reagents, 2009, 31(6), 401—404
	( 张俊, 岑涛, 王安明, 周成, 杜志强, 武秀明, 沈树宝. 化学试剂, 2009, 31(6), 401—404)
[35]	Cen T., Zhou C., Wu X. M., Jiang B., Zhu S. M., Shen S. B., Chinese J. Org. Chem., 2010, 30(6), 837—842
	( 岑涛, 周成, 武秀明, 姜波, 祝社民, 沈树宝. 有机化学, 2010, 30(6), 837—842)

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	—