25-羟基胆固醇及其类似物的合成与抗神经炎活性研究

doi:10.6023/cjoc202312002

摘要/Abstract

25-羟基胆固醇(25HC)具有许多重要的生物活性, 也是合成骨化二醇、骨化三醇等药物的关键中间体, 开发其原料易得、工艺简单以及易于放大的新合成路线具有重要意义. 以廉价易得的猪去氧胆酸为原料, 经羟基酯化、脱羧碘代、与α-酮酸酯反应进而水解脱羧、羟基磺酸酯化及消除, 最后与格氏试剂反应共6步反应合成了3个25-羟基胆固醇及其类似物, 总产率为20.8%~23.0%. 新路线操作简单、易于放大, 有望进一步发展成工业化合成路线. 通过改变酮酸酯及格氏试剂可方便地得到不同的25-羟基胆固醇类似物. 合成所得到的15个化合物中, 有9个是新的化合物. 对15个化合物进行生物活性研究, 结果表明, 26,27-二乙基胆甾-5-烯-3β,25-二醇(8c)具有良好的抗神经炎活性, 可通过抑制核转录因子(NF-κB)和丝裂原活化蛋白激酶(MAPK/JNK)信号通路, 抑制LPS(脂多糖)诱导的小鼠小胶质瘤细胞(BV-2)细胞释放NO、下调白细胞介素6 (IL-6)、肿瘤坏死因子α (TNF-α)、白细胞介素1β (IL-1β)等炎症因子的信使核糖核酸(mRNA)水平, 表明8c是有潜力的抗神经炎症的化合物.

关键词: 25-羟基胆固醇, 25-羟基胆固醇类似物, 合成, 抗神经炎活性

25-Hydroxycholesterol (25HC) has many important biological activities and is also a key intermediate for the synthesis of calcifediol, calcitriol, etc. It is of great significance to develop a new synthetic route which is easy to available raw materials, simple process, and easy to scale up. In this report, three 25-hydroxycholesterol and its analogues were synthesized from hyodeoxycholic acid via esterification of hydroxyl groups, decarboxylation iodization of carboxyl groups, ketonic hydrolysis after nucleophilic substitution with α-ketoester, sulfonate esterification then elimination, and finally reaction with Grignard reagent. The total yield was 20.8%~23.0%. The new route is simple to operate and easy to scale up, and is expected to be further developed into an industrial synthetic route. Similar analogues of 25-hydroxycholesterol can be easily prepared by changing α-ketoesters and Grignard reagents. Among the 15 compounds synthesized, there are 9 new compounds. Their biological activities were evaluated. The results showed that 26,27-diethylcholest-5-ene-3β,25-diol (8c) has good anti-neuritis activity and can inhibit NO release from LPS (lipopolysaccharides)-induced BV-2 (mouse microglia) cells by inhibiting Nuclear Factor kappa-B (NF-κB) and Mitogen-activated protein kinase/c-Jun N-terminal Kinase (MAPK/JNK) signaling pathways, and down-regulating mRNA-levels of inflammatory cytokines such as Interleukin-6 (IL-6), Tumor Necrosis Factor-α (TNF-α) and Interleukin-1β (IL-1β), indicating that 8c is a potential anti-neuroinflammatory compound.

Key words: 25-hydroxycholesterol, 25-hydroxycholesterol analogues, synthesis, anti-neuritis activity

引用此文

蓝柳淞, 杨倩, 李永怡, 方淑君, 黄宇轩, 苏骏成, 潘成学, 苏桂发. 25-羟基胆固醇及其类似物的合成与抗神经炎活性研究[J]. 有机化学, 2024, 44(7): 2305-2314.

Liusong Lan, Qian Yang, Yongyi Li, Shujun Fang, Yuxuan Huang, Juncheng Su, Chengxue Pan, Guifa Su. Synthesis and Anti-neuroinflammatory Activities of 25-Hydroxycholesterol and Its Analogues[J]. Chinese Journal of Organic Chemistry, 2024, 44(7): 2305-2314.

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图/表 8

图1. 合成25-羟基胆固醇的原料

Figure 1. Raw material for the synthesis of 25-hydroxy-chole- sterol

图式1. 本文报道的25-羟基胆固醇及其类似物的合成路线

Scheme 1. Synthesis of 25HC and its analogues reported in this paper

Compd.	Survival rate/%	Compd.	Survival rate/%
4a	102.64±5.45	6c	121.65±3.41***
4b	3.84±0.36****	7a	120.48±3.71***
4c	3.26±0.04****	7b	98.38±4.58
5a	54.69±17.96*	7c	118.12±3.15***
5b	4.06±0.57****	8a	27.52±0.69****
5c	4.12±0.51****	8b	23.97±3.76****
6a	104.09±4.36	8c	112.82±8.91
6b	120.31±4.51**	Control	100±2.56
HDCA	102.18±8.06	DEX	85.05±2.84***

表1. 化合物BV-2小胶质细胞存活率(%)的影响(mean±SE)a

Table 1. Effect of compounds on the survival rate (%) of BV-2 microglia (mean±SE)

Compd.	Release of nitric oxide/%	Compd.	Release of nitric oxide/%
4a	76.18±13.74*	7a	59.70±8.25**
6a	61.16±11.65**	7b	61.16±5.53***
6b	45.78±23.93*	7c	71.78±8.25**
6c	39.55±16.41**	8c	12.98±6.31****
Control	12.82±8.25	DEX	30.03±8.32***
LPS	100±7.20^####	HDCA	56.62±6.42

表2. 化合物对LPS激活BV-2小胶质细胞释放一氧化氮(%)的影响(mean±SE)a

Table 2. Effects of compounds on the release of nitric oxide (%) by LPS-activated BV-2 microglia (mean±SE)

Compd.	Survival rate/%	Compd.	Survival rate/%
4a	43.54±2.98****	7a	1.90±0.12****
6a	25.64±2.75****	7b	1.33±0.41****
6b	26.14±5.74****	7c	1.60±0.21****
6c	6.47±1.30****	8c	56.22±1.30****
Control	100±1.19	DEX	14.37±2.93****
LPS	88.57±2.01

表3. 化合物对LPS活化的BV-2小胶质细胞存活率(%)的影响(mean±SE)a

Table 3. Effect of compounds on the survival rate (%) of LPS-activated BV-2 microglia (mean±SE)

Compd.	Control	LPS	Release of nitric oxide/%				IC₅₀/(μmol•L^－1)
Compd.	Control	LPS	6.25 μmol•L^－1	12.50 μmol•L^－1	25.00 μmol•L^－1	50.00 μmol•L^－1	IC₅₀/(μmol•L^－1)
8c	10.68±4.56	100±11.10	63.81±5.74	39.69±7.72	17.29±4.75	11.26±5.47	9.55
DEX	10.68±4.56	100±11.10	—	—	—	58.36±4.97	—

表4. 化合物8c对LPS激活BV-2小胶质细胞释放一氧化氮(%)的影响(mean±SE)a

Table 4. Effects of compound 8c on the release of nitric oxide (%) by LPS-activated BV-2 microglia (mean±SE)

图2. 化合物8c对LPS诱导的BV-2细胞IL-6、TNF-α和IL-1β的mRNA表达量的影响

Figure 2. Effects of compound 8c on mRNA expression of IL-6, TNF-α and IL-1β in LPS-induced BV-2 cells Error bars represent s.d., *p<0.05, **p<0.01, ***p<0.001 compared to LPS group. ####p<0.0001 compared to control group.

图3. 化合物8c通过抑制NF-κB和MAPK/JNK信号通路的激活抑制LPS诱导的炎症反应

Figure 3. Compound 8c inhibits LPS-induced inflammation by inhibiting the activation of NF-κB and MAPK/JNK signaling pathways

参考文献 21

[1]	Cyster, J. G.; Dang, E. V.; Reboldi, A.; Yi, T.-S. Nat. Rev. Immunol. 2014, 14, 731.
[2]	Adams, C. M.; Reitz, J.; De Brabander, J. K.; Feramisco, J. D.; Li, L.; Brown, M. S.; Goldstein, J. L. J. Biol. Chem. 2004, 279, 52772.
[3]	Cashikar, A. G.; Toral-Rios, D.; Timm, D.; Romero, J.; Strickland, M.; Long, J. M.; Han, X.-L.; Holtzman, D. M.; Paul, S. M. J. Lipid Res. 2023, 64, 100350.
[4]	a) Reboldi, A.; Dang, E. V.; McDonald, J. G.; Liang, G.-S.; Russell, D. W.; Cyster, J. G. Science 2014, 345, 679. doi: 10.1126/science.1254790 pmid: 25104388
	(b) Jang, J.; Park, S.; Hur, H. J.; Cho, H.-J.; Hwang, I.; Kang, Y. P.; Im, I.; Lee, H.; Lee, E.; Yang, W.; Kang, H.-C.; Kwon, S. W.; Yu, J.-W.; Kim, D.-W. Nat. Commun. 2016, 7, 13129. pmid: 25104388
	(c) Civra, A.; Cagno, V.; Donalisio, M.; Biasi, F.; Leonarduzzi, G.; Poli, G.; Lembo, D. Sci. Rep. 2014, 4, 7487. pmid: 25104388
[5]	a) Liu, S.-Y.; Aliyari, R.; Chikere, K.; Li, G.-M.; Marsden, M.D.; Smith, J. K.; Pernet, O.; Guo, H.-T.; Nusbaum, R.; Zack, J. A.; Freiberg, A. N.; Su, L.-S.; Lee, B.; Cheng, G.-H. Immunity 2013, 38, 92. doi: 10.1016/j.immuni.2012.11.005 pmid: 23273844
	(b) Mao, S.; Ren, J.; Xu, Y.; Lin, J.; Pan, C.; Meng, Y.; Xu, N. Eur. J. Pharmacol. 2022, 926, 175033. pmid: 23273844
	(c) Chen, Y.; Wang, S.; Yi, Z.; Tian, H.; Aliyari, R.; Li, Y.; Chen, G.; Liu, P.; Zhong, J.; Chen, X.; Du, P.; Su, L.; Qin, F. X.-F.; Deng, H.; Cheng, G. Sci. Rep. 2014, 4, 7242. pmid: 23273844
[6]	a) Lu, Z.; McBrearty, N.; Chen, J.-Y.; Tomar, V. S.; Zhang, H.-R.; De Rosa, G.; Tan, A.-W.; Weljie, A. M.; Beiting, D. P.; Miao, Z.; George, S. S.; Berger, A.; Saggu, G.; Diehl, J. A.; Koumenis, C.; Fuchs, S. Y. Cell Metab. 2022, 34, 1342.
	(b) You, J.-S.; Lim, H.; Kim, T.-H.; Oh, J.-S.; Lee, G.-J.; Seo, Y.-S.; Kim, D. K.; Yu, S.-K.; Kim, H.-J.; Kim, C. S.; Kim, J.-S. Anticancer Res. 2020, 40, 779
[7]	a) Cao, Q.; Luo, J.; Xiong, Y.; Liu, Z.-Z.; Ye, Q.-F. Int. Immunopharmacol. 2021, 96, 107643.
	(b) Lv, S.-Y.; Ju, C.-H.; Peng, J.-T.; Liang, M.-L.; Zhu, F.; Wang, C.; Huang, K.; Cheng, M.; Zhang, F.-X. Int. J. Biol. Sci. 2020, 16, 298.
[8]	Zhang, L.; Sun, B.; Su, W.-K.; Jin, C. Chin. Pharm. J. 2019, 50, 695 (in Chinese).
	(张凉, 孙彬, 苏为科, 金灿, 中国医药工业杂志, 2019, 50, 695.)
[9]	Zhao, Q.; Ji, L.; Qian, G.-P.; Liu, J.-G.; Wang, Z.-Q.; Yu, W.-F.; Chen, X.-Z. Steroids 2014, 85, 1. doi: 10.1016/j.steroids.2014.02.002 pmid: 24582707
[10]	a) Ogawa, S.; Kakiyama, G.; Muto, A.; Hosoda, A.; Mitamura, K.; Ikegawa, S.; Hofmann, A. F.; Iida, T. Steroids 2009, 74, 81.
	(b) Wang, J.-T.; Chen, W.-Z.; Xia, J.; Zhao, P.; Xie, J.; Tang, L. Chem. React. 2017, 39, 668 (in Chinese).
	(王建塔, 陈文章, 夏晶, 赵平, 谢珺, 汤磊, 化学试剂, 2017, 39, 668.)
[11]	a) Miyamoto, K.; Kubodera, N.; Murayama, E.; Ochi, K.; Mori, T.; Matsunaga, I. Synth. Commun. 1986, 16, 513. pmid: 28300584
	(b) Brownholland, D. P.; Covey, D. F. Steroids 2017, 121, 22. doi: S0039-128X(17)30037-5 pmid: 28300584
[12]	a) Liu, S.; Wang, G.-Q.; Liu, Z.-K.; Wang, Q.-A. Chin. J. Org. Chem. 2013, 33, 2216 (in Chinese).
	(刘双, 汪钢强, 刘张坤, 汪秋安, 有机化学, 2013, 33, 2216.)
	(b) Wang, Z.-Q.; Jiang, L.-Z.; Rong, S.-H. Chin. J. Med. Chem. 1994, 4, 89 (in Chinese).
	(王钟麒, 姜立中, 容士宏, 中国药物化学杂志, 1994, 4, 89.)
	(c) Wang, Z.-Q.; Yuan, B.-F. Acta Chim. Sinica 1994, 52, 403 (in Chinese).
	(王钟麒, 阮奔放, 化学学报, 1994, 52, 403.)
	(d) Ruan, B.-F.; Wang, Z.-Q. Acta Chim. Sinica 1993, 51, 612 (in Chinese).
	(阮奔放, 王钟麒, 化学学报, 1993, 51, 612.)
	(e) Wang, Z.-Q.; Jiang, L.-Z.; Zhou, W.-S.; Gong, Y.-M.; Qian, Y. Sci. China, Ser. B: Chem., ife Sci., Earth Sci. 1991, 34, 680 (in Chinese).
	(王钟麒, 姜立中, 周维善, 龚逸民, 钱涌, 中国科学,
	(B辑, 化学. 生命科学, 地学), 1991, 34, 680.)
	(f) Wang, Z.-Q.; Jiang, L.-Z.; Zhou, W.-S. Acta Chim. Sinica 1990, 48, 99 (in Chinese).
	(王钟麒, 姜立中, 周维善, 化学学报, 1990, 48, 99.)
	(g) Zhou, W.-S.; Wang, Z.-Q.; Jiang, B. Acta Chim. Sinica 1988, 46, 1150 (in Chinese).
	(周维善, 王钟麒, 姜标, 化学学报, 1988, 46, 1150.)
[13]	a) Peng, Y.-H.; Wen, Y.-C.; Chen, Y.-Q. Acta Chim. Sinica 1985, 43, 698 (in Chinese).
	(彭逸华, 温业淳, 陈毓群, 化学学报, 1985, 43, 698.)
	(b) Wang, Z.-Q.; Jiang, L.-Z.; Zhou, W.-S. Chin. Chem. Lett. 1992, 3, 409.
	(c) Jin, C.; Wang, Y.-L.; Sun, B.; Su, W.-K. J. Chem. Res. 2018, 42, 96.
[14]	Nakai, Y.; Moriyama, K.; Togo, H. Eur. J. Org. Chem. 2016, 2016, 768.
[15]	a) Tan, X. Q.; Song, T.; Wang, Z. T.; Chen, H.; Cui, L.; Li, C. Z. Org. Lett. 2017, 19, 1634.
	(b) Wang, Z. T.; Zhu, L.; Yin, F.; Su, Z. Q.; Li, Z. D.; Li, C. Z. J. Am. Chem. Soc. 2012, 134, 425.
[16]	Camps, P.; Lukach, A. E.; Pujol, X.; Vazquez, S. Tetrahedron 2000, 56, 2703.
[17]	Candish, L.; Standley, E. A.; Gomez-Suarez, A.; Mukherjee, S.; Glorius, F.Chem.-Eur. J. 2016, 22, 9971.
[18]	Partridge, J. J.; Faber, S.; Uskokovic, M. R. Helv. Chim. Acta 1974, 57, 84.
[19]	Xiao, J.; Song, J.-Y.; Lin, B.; Li, W.; Yang, Y.-Q.; Liu, J.-Y.; Hou, Y.; Chen, G.; Li, N. J. Nat. Prod. 2020, 83, 864.
[20]	Liu, Y.-C.; Feng, N.; Li, W.-W.; Tu, P.-F.; Chen, J.-P.; Han, J.-Y.; Zeng, K.-W. Molecules 2020, 25, 2840.
[21]	Eguchi, T.; Sai, H.; Takatsuto, S.; Hara, N.; Ikekawa, N. Chem. Pharm. Bull. 1988, 36, 2303.