五环三萜葡萄糖缀合物的设计、合成及体外抗流感病毒活性研究

doi:10.6023/cjoc202111008

摘要/Abstract

前期研究发现齐墩果酸(OA)的C28位葡萄糖偶联物在细胞水平具有显著抗流感病毒的活性, 但不同五环三萜母核偶联葡萄糖抗流感病毒活性的影响并未阐明. 本研究采用不同五环三萜母核, 通过Cu(I)催化的炔烃-叠氮化物环加成反应(CuAAC), 与葡萄糖偶联制备得到化合物物4c~8d, 通过体外抗流感病毒A/WSN/33 (H1N1)活性研究, 发现化合物4d具有显著的抗流感病毒活性(IC₅₀为9.09 μmol/L), 且对MDCK (Madin-Darbey Canine Kidney)细胞无明显毒性(CC₅₀>100 μmol/L). 构效关系研究明确了葡萄糖模块以及五环三萜母核对化合物抗流感病毒活性的提升均具有重要作用. 本研究进一步完善了五环三萜及其衍生物抗流感病毒的构效关系, 为这类天然产物抗病毒的深入研究奠定了基础.

关键词: 五环三萜, 流感病毒, 进入抑制剂, 构效关系

Previous studies have revealed that oleanolic acid (OA) C28-glucose conjugate displayed potent anti-influenza viral activity in vitro. However, the effect of pentacyclic triterpene scaffold coupled with glucose on anti-influenza virus activity has not been clarified. Herein, as a continuous further study, a series of pentacyclic triterpene-glucose conjugates were prepared via CuAAC reaction, and the anti-influenza activity of these compounds was evaluated in vitro. Among them, compound 4d, an betulinic acid (BA)-glucose conjugate, showed potent anti-influenza virus activity (IC₅₀ value of 9.09 μmol/L), and no obvious cytotoxic effect on MDCK (Madin-Darbey Canine Kidney) cells was observed (CC₅₀>100 μmol/L). Structure-activity relationship studies showed that both triterpene scaffolds and glucose moiety have important effects on the anti-viral activity. Overall, this finding further extends the utility of triterpene derivatives in anti-influenza virus drug design.

Key words: pentacyclic triterpene, influenza virus, entry inhibitors, structure-activity relationship

引用此文

蔡铭, 邵亮, 杨帆, 张继虹, 俞飞. 五环三萜葡萄糖缀合物的设计、合成及体外抗流感病毒活性研究[J]. 有机化学, 2022, 42(5): 1453-1462.

Ming Cai, Liang Shao, Fan Yang, Jihong Zhang, Fei Yu. Design, Synthesis of Pentacyclic Triterpenoid Glucose Conjugate and in vitro Activity against Influenza Virus[J]. Chinese Journal of Organic Chemistry, 2022, 42(5): 1453-1462.

导出引用 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

分享此文

图/表 7

图1. 齐墩果酸及其衍生物的结构式

Figure 1. Structural formulas of oleanolic acid and its derivatives

图式1. 白桦脂酸葡萄糖缀合物的合成

Scheme 1. Synthesis of betulinic acid (BA)-glucose conjugate Reagents and conditions: (a) TBTU, DIPEA, THF, r.t.; (b) 2-propynylamine, Na2CO3, DMF, r.t., 2 h; (c) CuSO4•5H2O, Na-L-ascorbate, CH2Cl2/H2O, r.t.; (d) MeONa, MeOH, r.t., 3 h

化合物结构式	化合物		细胞存活率/%		流感病毒抑制率/%
	4		106.12±8.38		59.00±10.18
	4c		91.27±0.03		8.46±23.27
	4d		106.53±9.50		92.94±1.45
	5		4.08±0.34		99.48±1.08
	5c		110.24±0.002		NT
	5d		105.15±11.41		54.16±11.54
	6		3.98±1.90		95.14±7.36
	6c		56.04±0.008		58.51±4.72
	6d		74.02±6.92		80.65±16.47
化合物结构式		化合物		细胞存活率/%		流感病毒抑制率/%
		7		3.30±0.06		99.85±0.31
		7c		103.68±0.02		10.43±5.20
		7d		4.18±1.45		97.41±3.72
		8		102.05±10.33		70.99±14.04
		8c		73.23±3.62		96.62±15.17
		8d		101.63±6.89		91.68±8.14

表1. 五环三萜葡萄糖缀合物结构式及抗流感病毒活性初步评价结果($\overline{x}\pm \text{s},$n=3)a

Table 1. Structural formula of pentacyclic triterpene-glucose conjugate and preliminary evaluation results of its anti-influenza virus activity ($\overline{x}\pm \text{s},$n=3)

Compound	CC₅₀^a/(μmol•L^–1)	IC₅₀^b/(μmol•L^–1)	SI^c
OSV-p^d	>100	1.82	>54.59
2^d	>100	5.47	>18.28
4d	>100	9.09	>10.99
8d	>100	12.56	>7.96

表2. 五环三萜葡萄糖缀合物抗A/WSN/33 (H1N1)活性($\overline{x}\pm \text{s},$ n=3)

Table2. Activity of the pentacyclic triterpene-glucose conjugate anti- influenza virus A/WSN/33 ($\overline{x}\pm \text{s},$ n=3)

图2. (A)化合物4d有效抑制A/WSN/33诱导的细胞病变; (B)化合物4d和血凝素抗体可以抑制A/WSN/33引起的鸡红细胞凝集 Figure 2 (A) Compound 4d can effectively inhibit CPE induced by A/WSN/33; (B) Compound 4d and HA antibody can inhibit hemagglutination caused by A/WSN/33

Compd.	Binding energy^a/(kJ•mol^–1)	Chemical feature^b
4c	–22.01	CHB^c, π-Alkyl
5c	–14.69	CHB, π-DHB^d, Alkyl
6c	–22.30	CHB, Alkyl
7c	–21.46	CHB, π-DHB, π-Alkyl
8c	–21.42	CHB, π-Sigma, π-Alkyl
4d	–24.98	CHB, π-Cation, π-Alkyl
5d	–21.55	CHB, π-Anion, π-Alkyl
6d	–22.93	CHB, π-Cation, π-Alkyl
7d	–22.55	CHB, π-Alkyl
8d	–23.10	CHB, π-Anion, Alkyl

表3. 五环三萜葡萄糖缀合物与血凝素蛋白最低自由结合能与功能基团特征

Table 3. Minimum free binding energy and chemical feature of pentacyclic triterpene-glucose conjugates binding to HA protein

图3. 化合物4d与血凝素蛋白分子对接模拟: (A)化合物4d在血凝素蛋白中的结合位点; (B)化合物4d与血凝素蛋白相互作用的三维模型图; (C)化合物4d与血凝素蛋白相互作用的二维模型图

Figure 3. Molecular docking simulation of compound 4d targeting HA protein: (A) the binding site of compound 4d in HA protein; (B) the 3D model diagram of the interaction between compound 4d and HA protein; (C) the 2D model diagram of the interaction between compound 4d and HA protein

参考文献 18

[1]	Li, H.; Li, M.; Xu, R.; Wang, S.; Zhang, Y.; Zhang, L.; Zhou, D.; Xiao, S. Eur. J. Med. Chem. 2019, 163, 560. doi: 10.1016/j.ejmech.2018.12.006
[2]	Chintakrindi, A. S.; Gohil, D. J.; Chowdhary, A. S.; Kanyalkar, M. A. Bioorg. Med. Chem. 2020, 28, 115191. doi: 10.1016/j.bmc.2019.115191
[3]	Cline, T. D.; Karlsson, E. A.; Seufzer, B. J.; Schultz-Cherry, S. J. Virol. 2013, 87, 1411. doi: 10.1128/JVI.02682-12 pmid: 23152519
[4]	Chintakrindi, A. S.; Gohil, D. J.; Kothari, S. T.; Chowdhary, A. S.; Kanyalkar, M. A. Med. Chem. Res. 2018, 27, 1013. doi: 10.1007/s00044-017-2124-2
[5]	Naesens, L.; Stevaert, A.; Vanderlinden, E. Curr. Opin. Pharmacol. 2016, 30, 106. doi: S1471-4892(16)30088-1 pmid: 27570127
[6]	Lee, N.; Hurt, A. C. Curr. Opin. Infect. Dis. 2018, 31, 520. doi: 10.1097/QCO.0000000000000498
[7]	Yen, H. L.; Hoffmann, E.; Taylor, G.; Scholtissek, C.; Monto, A. S.; Webster, R. G.; Govorkova, E. A. J. Virol. 2006, 80, 8787. doi: 10.1128/JVI.00477-06
[8]	Tian, Z.; Si, L.; Meng, K.; Meng, K.; Zhou, X.; Zhang, Y.; Zhou, D.; Xiao, S. Eur. J. Med. Chem. 2017, 134, 133. doi: 10.1016/j.ejmech.2017.03.087
[9]	Yu, M.; Si, L.; Wang, Y.; Wu, Y.; Yu, F.; Jiao, P.; Shi, Y.; Wang, H.; Xiao, S.; Fu, G.; Tian, K.; Wang, Y.; Guo, Z.; Ye, X.; Zhang, L.; Zhou, D. J. Med. Chem. 2014, 57, 10058. doi: 10.1021/jm5014067
[10]	Luo, M.; Wu, X.; Li, Y.; Guo, F. Molecules 2021, 26, 895. doi: 10.3390/molecules26040895
[11]	Su, Y.; Meng, L.; Sun, J.; Li, W.; Shao, L.; Chen, K.; Zhou, D., Yang, F.; Yu, F. Eur. J. Med. Chem. 2019, 182, 111622. doi: 10.1016/j.ejmech.2019.111622
[12]	Shao, L.; Yang, F.; Li, W. J.; Yu, F. Chin. J. Org. Chem. 2021, 41, 2454. (in Chinese) doi: 10.6023/cjoc202010029
	(邵亮, 杨帆, 李维嘉, 俞飞, 有机化学, 2021, 41, 2454.)
[13]	Nouri-Felekori, M.; Nezafati, N.; Moraveji, M.; Hesaraki, S.; Ramezani, T. Int. J. Biol. Macromol. 2021, 183, 2030. doi: 10.1016/j.ijbiomac.2021.06.005 pmid: 34097959
[14]	Agrahari, A. K.; Bose, P.; Jaiswal, M. K.; Rajkhowa, S.; Singh, A. S.; Hotha, S.; Mishra, N.; Tiwari, V. K. Chem. Rev. 2021, 121, 7638. doi: 10.1021/acs.chemrev.0c00920 pmid: 34165284
[15]	Devaraj, N. K.; Finn, M. G. Chem. Rev. 2021, 121, 6697. doi: 10.1021/acs.chemrev.1c00469 pmid: 34157843
[16]	Chen, D.; Zhang, P.; Sun, Y.; Wang, P.; Zhang, C.; Kong, L.; Zhang, J.; Sun, H.; Wen, X. Org. Biomol. Chem. 2016, 14, 11154. doi: 10.1039/C6OB02126C
[17]	Li, H.; Li, M.; Xu, R.; Wang, S.; Zhang, Y.; Zhang, L.; Zhou, D.; Xiao, S. Eur. J. Med. Chem. 2019, 163, 560. doi: 10.1016/j.ejmech.2018.12.006
[18]	Spackman, E.; Sitaras, I. Methods. Mol. Biol. 2020, 2123, 11. doi: 10.1007/978-1-0716-0346-8_2 pmid: 32170677