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中国药学(英文版) ›› 2016, Vol. 25 ›› Issue (7): 517-525.DOI: 10.5246/jcps.2016.07.057

• 【研究论文】 • 上一篇    下一篇

大鼠血浆中新型α7-烟碱型乙酰胆碱受体正向变构调节剂LD486的液相色谱-质谱串联测定方法学的建立及其药代动力学特征

黄晓敏1,3, 焦文宣2, 孙崎2,3*, 王克威1,3*   

  1. 1. 北京大学医学部 药学院 分子与细胞药理学系, 北京 100191
    2. 北京大学医学部 药学院 化学生物学系, 北京 100191
    3. 北京大学医学部 天然药物及仿生药物国家重点实验室, 北京 100191
  • 收稿日期:2016-04-11 修回日期:2016-05-04 出版日期:2016-07-19 发布日期:2016-05-20
  • 通讯作者: Tel.: +86-010-82805605, E-mail: sunqi@bjmu.edu.cn, wangkw@bjmu.edu.cn
  • 基金资助:

    Research grants to KeWei Wang from Ministry of Science and Technology of China (Grant Nos. 2013CB531302, 2014ZX09507003-006-004, and 2013ZX09103001-015), and National Natural Science Foundation of China (Grant No. 21572011).

Pharmacokinetic characterization of a novel α7 nicotinic acetylcholine receptor (nAChR) positive allosteric modulator LD486 in rat plasma using a validated LC-MS/MS assay

Xiaomin Huang1,3, Wenxuan Jiao2, Qi Sun2,3*, Kewei Wang3*   

  1. 1. Department of Molecular and Cellular Pharmacology, School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing 100191, China
    2. Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing 100191, China
    3. State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing 100191, China
  • Received:2016-04-11 Revised:2016-05-04 Online:2016-07-19 Published:2016-05-20
  • Contact: Tel.: +86-010-82805605, E-mail: sunqi@bjmu.edu.cn, wangkw@bjmu.edu.cn
  • Supported by:

    Research grants to KeWei Wang from Ministry of Science and Technology of China (Grant Nos. 2013CB531302, 2014ZX09507003-006-004, and 2013ZX09103001-015), and National Natural Science Foundation of China (Grant No. 21572011).

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摘要：

神经型α7-烟碱型乙酰胆碱受体功能的不足与认知缺陷以及多种神经精神障碍相关。在动物模型中, 激活α7-烟碱型乙酰胆碱受体可以提高学习记忆能力以及感觉门控。为了增强新型的功能, 我们通过筛选发现了激动α7-烟碱型乙酰胆碱受体的正向调节剂LD486。在本研究中, 我们建立了一个简单可靠且高效的液相色谱质谱联用的方法(LC-MS/MS)并分析了这种新型的正向变构调节剂LD486在大鼠血浆中的浓度以及药代动力学参数。含有LD486的血浆样品用反向C18色谱柱分离, 流动相为乙腈和0.1%甲酸水溶液(80:20, v/v), 流速为0.5 mL/min, 内标为A1223。液相流出液使用电喷雾离子源(ESI, 正离子模式), 质谱多反应监测模式检测, 检测离子: LD486为m/z 419.0 [M+H]+→m/z 169.0, A1223为m/z 400.0 [M+H]+→m/z 129.9。LD486在4–4000 ng/mL范围内, 线性关系良好, 最低定量限为4 ng/mL。在最低定量限时, 依然有满意的日内日间精密度, 提取回收率, 以及基质效应。稳定性测试提示血浆中LD486在反复冻融, 22 ºC室温下放置2小时和12小时, 以及–20 ºC保存2周均保持稳定。我们将此方法成功地应用到测定大鼠血浆中LD486静脉注射(1 mg/kg)以及口服(3 mg/kg和30 mg/kg)后给药途径的药代动力学特征。

关键词: 乙酰胆碱受体正向变构调节剂, LD486, 液相色谱质谱联用, 药代动力学, 大鼠血浆, α7受体通道

Abstract:

The deficiency of neuronal α7-nicotinic acetylcholine receptor (α7 nAChR) channel is implicated in cognition deficit and neuropsychiatric disorders. Chemical activation of α7 nAChR improves learning, memory, and sensory gating in animal models. To identify novel α7 nAChR activators, we recently identified a lead compound LD486 that as a positive allosteric modulator (PAM) selectively activates α7 nAChR. In this study we developed a simple, reliable and efficient assay of high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) and analyzed the pharmacokinetics of this novel LD486 in rat plasma. Separation of compound LD486 from plasma was achieved using a reversed-phase C18 column with a mobile phase of 0.1% formic acid in H2O (80:20, v:v), using compound A1223 as an internal standard (IS) with a flow rate of 0.5 mL/min. The quantitative analysis was performed using multiple reaction monitoring (MRM) at the specific ion transition of m/z 419.0 [M+H]+→m/z 169.0 for LD486 and m/z 400.0 [M+H]+→m/z 129.9 for A1223 in the positive ion mode with electrospray ionization (ESI) source. This validated LC-MS/MS method shows good linearity over the range 4–4000 ng/mL with the lowest limit of quantitation (LLOQ) at 4 ng/mL as well as satisfied intra- and inter-day precision, accuracy, extraction recovery and matrix effect.The stability testing indicates that LD486 in rat blood is stable under three freeze-thaw cycles, in room temperature at 22 oC for 2 h and 12 h, and for 2 weeks at −20 oC. This developed method was successfully applied to the pharmacokinetic analysis of intravenous (1 mg/kg) and oral (3 mg/kg and 30 mg/kg) administration of LD486 in SD rats.

Key words: AChR PAM, LD486, LC-MS/MS, Pharmacokinetics, Rat blood, α7

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