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浙江大学学报(医学版)  2022, Vol. 51 Issue (4): 415-421    DOI: 10.3724/zdxbyxb-2022-0174
专题报道     
长链亲脂性氮氧自由基HPN衍生物的设计合成及对缺氧小鼠存活的影响
达清越1,2,张洁3,张朋朋3,石志群3,马慧萍3,*,景临林2,*
1. 兰州大学药学院,甘肃 兰州 730000
2. 西安交通大学第一附属医院脑科学研究中心,陕西 西安 710061
3. 中国人民解放军联勤保障部队第九四〇医院药剂科 全军高原医学实验室,甘肃 兰州 730050
Design, synthesis and anti-hypoxia activity of HPN derivatives containing lipophilic long chains
DA Qingyue1,2,ZHANG Jie3,ZHANG Pengpeng3,SHI Zhiqun3,MA Huiping3,*,JING Linlin2,*
1. School of Pharmacy, Lanzhou University, Lanzhou 730000, China;
2. Center for Brain Science, the First Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710061, China;
3. Department of pharmacy, the 940th Hospital of Joint Logistics Support Force of Chinese People’s Liberation Army, Key Laboratory of the Plateau Medicine, Lanzhou 730050, China
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摘要:

目的:设计合成长链取代的2-(4′-羟基苯基)-4,4,5,5-四甲基咪唑烷-1-氧基-3-氧化物(HPN)衍生物,以增强HPN的抗缺氧活性。 方法:以乙腈为溶剂、碳酸钾为缚酸剂, 60?℃反应条件下将HPN分别与6-溴己醇、6-溴己酸乙酯和6-溴己烷进行烷基化反应得到3个长链亲脂性HPN衍生物1、3和5;衍生物1在氢氧化钠甲醇/水体系下经过水解反应得到衍生物2;以二氯甲烷为溶剂, NN′-二异丙基碳二亚胺为脱水剂,HPN与己酸进行酯化反应,得到衍生物4。采用红外光谱、电子顺磁共振和高分辨质谱表征衍生物1~5的结构,高效液相色谱法检测衍生物的纯度,计算油水分配系数(log P值)评估衍生物的脂溶性,并通过缺氧小鼠常压密闭缺氧实验和急性减压耐受实验考察HPN及衍生物1~5的抗缺氧活性。 结果:5个衍生物的结构经红外光谱、电子顺磁共振和高分辨质谱分析确认,产率均在92%以上,纯度均在96%以上。衍生物1~5的log P值分别为2.78、2.00、2.04、2.88和3.10,均高于HPN(0.97);在给药剂量为 0.3?mmol/kg时均能显著延长常压密闭缺氧小鼠的生存时间,并将急性减压缺氧小鼠的死亡率分别下降至60%、70%、60%、70%、40%。 结论:衍生物1~5的合成方法简便,产率较高,且在剂量较小的情况下,仍然表现出类似或优于HPN的抗缺氧活性,其中衍生物5的活性最佳。

关键词: 氮氧自由基2-(4′-羟基苯基)-4,4,5,5-四甲基咪唑烷-1-氧基-3-氧化物亲脂性长链合成抗缺氧小鼠    
Abstract:

Objective: To design and synthesize long-chain substituted 2-[(4′-hydroxyethoxy) phenyl]-4,4,5,5-tetramethyl-2-imidazoline-1-oxyl 3-oxide (HPN) derivatives with enhanced anti-hypoxic activity. Methods: HPN derivatives 1, 3, 5 containing lipophilic long chains were synthesized via the alkylation of HPN with 6-bromohexan-1-ol, ethyl 6-bromohexanoate or 6-bromohexane, respectively using acetonitrile as the solvent and K 2CO 3 as the acid-binding agent at 60?℃. Derivative 2 was synthesized via hydrolysis reactions of derivative 1 in the NaOH/CH 3OH/H 2O system. Using dichloromethane as the solvent and N, N′-diisopropylcarbodiimide as the dehydrating agent, HPN underwent esterification with hexanoic acid to obtain derivative 4. The structures of derivatives 1-5 were characterized by infrared spectroscopy, electron paramagnetic resonance and high resolution mass spectrometry. The purities of derivatives were detected by high performance liquid chromatography, and the lipid solubilities of derivatives were evaluated by calculating the oil-water partition coefficients (log P). Anti-hypoxia activities of HPN and its long-chain lipophilic derivatives 1-5 were evaluated using normobaric hypoxia test and acute decompression hypoxia test. Results: The structures of the derivatives were confirmed by infrared spectroscopy, electron paramagnetic resonance and high resolution mass spectroscopy. The yields of target derivatives were all above 92%, and the purities were all above 96%. The log P values of derivatives 1-5 were 2.78, 2.00, 2.04, 2.88 and 3.10, which were higher than that of HPN (0.97). Derivatives 1-5 significantly prolonged the survival time of mice at the dose of 0.3?mmol/kg in normobaric hypoxic test and reduced the mortality rate of acute decompression hypoxic mice to 60%, 70%, 60%, 70% and 40%, respectively. Conclusions: The synthesis of derivatives 1-5 is convenient, and the yield is high. The synthesized derivatives especially derivative 5 show anti-hypoxic activity similar to or better than HPN at lower doses.

Key words: Nitronyl nitroxide    2-[(4′-Hydroxyethoxy) phenyl]-4,4,5,5-tetramethyl-2-imidazoline-1-oxyl 3-oxide    Lipophilic    Long chain    Synthesis    Anti-hypoxia    Mouse
收稿日期: 2022-04-16 出版日期: 2022-11-16
CLC:  R914.4  
基金资助: 国家自然科学基金(81202458); 全军医药卫生科研基金(CLZ12JA04); 中国博士后科学基金(2012M521926)
通讯作者: 马慧萍,景临林   
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引用本文:

达清越,张洁,张朋朋,石志群,马慧萍,景临林. 长链亲脂性氮氧自由基HPN衍生物的设计合成及对缺氧小鼠存活的影响[J]. 浙江大学学报(医学版), 2022, 51(4): 415-421.

DA Qingyue,ZHANG Jie,ZHANG Pengpeng,SHI Zhiqun,MA Huiping,JING Linlin. Design, synthesis and anti-hypoxia activity of HPN derivatives containing lipophilic long chains. J Zhejiang Univ (Med Sci), 2022, 51(4): 415-421.

链接本文:

https://www.zjujournals.com/med/CN/10.3724/zdxbyxb-2022-0174        https://www.zjujournals.com/med/CN/Y2022/V51/I4/415

图1  HPN衍生物1~5的合成路线 DIC: , ′-二异丙基碳二亚胺;DMAP:4-二甲氨基吡啶.

衍生物

质量(mg)

性 状

产率(%)

熔点(℃)

红外光谱波数(cm –1)

电子顺磁共振

高分辨质谱(m/s)

衍生物1

720

深蓝色固体

93

57.7~59.0

2986、2935、1720、1606、1490、1363、1311、1300、1254、1192、1012、828

五重峰, g=2.0082, a=7.74?G

理论值:C 21H 31N 2O 5{[M+H] +}392.2311;测量值:392.3171

衍生物2

345

深蓝色固体

95

166.8~168.8

2945、2869、1730、1604、1494、1379、1300、1260、1187、1035、838

五重峰, g=2.0082, a=7.74?G

理论值:C 19H 27N 2O 5{[M+H] +}364.1998;测量值:364.2844

衍生物3

330

深蓝色固体

95

135.1~136.8

3432、2941、2865、1605、1481、1360、1299、1258、1186、1030、833

五重峰, g=2.0082, a=7.74?G

理论值:C 19H 29N 2O 4{[M+H] +}350.2206;测量值:350.3041

衍生物4

320

深蓝色固体

92

72.7~73.6

2933、2852、1768、1601、1485、1390、1363、1300、1175、1100、1073、843

五重峰, g=2.0082, a=7.74?G

理论值:C 19H 27N 2O 4{[M+Na] +} 370.1854;测量值:370.1863

衍生物5

317

深蓝色固体

95

68.4~69.0

2937、2868、1604、1531、1485、1391、1360、1253、1184、1022、837

五重峰, g=2.0082, a=7.74?G

理论值:C 19H 29N 2O 3{[M+Na] +} 356.2076;测量值:356.2059

表1  HPN衍生物1~5的结构表征
图2  HPN衍生物1~5的高效液相色谱图谱 A:衍生物1;B:衍生物2;C:衍生物3;D:衍生物4;E:衍生物5. HPN:2-(4′-羟基苯基)-4,4,5,5-四甲基咪唑烷-1-氧基-3-氧化物; :保留时间.

组 别

n

生存时间(min)

延长率(%)

模型对照组

10

36.5±3.1

HPN组

10

46.1±4.8 **

26.5

衍生物1组

10

50.4±5.4 **

38.3

衍生物2组

10

47.6±4.7 **

30.6

衍生物3组

10

49.6±4.7 **

36.1

衍生物4组

10

45.7±5.1 **

25.3

衍生物5组

10

58.4±6.1 **#

60.3

表2  HPN衍生物1~5对常压密闭缺氧小鼠生存时间的影响
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