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

doi:10.3724/zdxbyxb-2022-0174

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 Qingyue^1,²,ZHANG Jie³,ZHANG Pengpeng³,SHI Zhiqun³,MA Huiping^3,*,JING Linlin^2,*

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， N， N′-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 或 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.

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.

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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