Page 51 - 中国药房2021年11期

Foretinib衍生物LWK-126在不同种属肝微粒体中的代谢稳定性

研究 Δ

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王忠元 ，杨华丽 ，崔 杏 ，朱高峰 ，汤 磊 ，廖伟科 （1.贵州省人民医院药剂科，贵阳 550002；2.贵州省化
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学合成药物研发利用工程技术研究中心，贵阳 550004）
中图分类号 969.1；R917 文献标志码 A 文章编号 1001-0408（2021）11-1325-05
DOI 10.6039/j.issn.1001-0408.2021.11.08
摘 要 目的：建立测定肝微粒体中Foretinib衍生物LWK-126质量浓度的方法，研究其在大鼠、比格犬、人肝微粒体中的代谢稳
定性。方法：采用体外肝微粒体孵育体系，将LWK-126分别溶于由还原型烟酰胺腺嘌呤二核苷酸磷酸溶液启动的大鼠、比格犬、
人肝微粒体孵育体系中，于37 ℃水浴下分别孵育0、5、10、20、30、60 min时加入含内标（1 μg/mL甲苯磺丁脲）的乙腈终止反应，并
采用超高效液相色谱-串联质谱法检测各孵育体系中LWK-126的质量浓度。以Waters BEH C18为色谱柱，以水（含0.01％甲酸）-乙
腈（含0.01％甲酸）为流动相进行梯度洗脱，流速为0.4 mL/min，柱温为30 ℃，进样量为2 μL；离子源为电喷雾电离源，采用正离子
模式采集，扫描范围为m/z 50→1 200。以孵育0 min时LWK-126的质量浓度为参照，计算其在不同孵育体系中的剩余百分率与酶
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动力学参数。结果：LWK-126检测质量浓度的线性范围为0.05～15 μg/mL（R ＝0.999 2），定量下限为0.05 μg/mL，最低检测限为
0.01 μg/mL，精密度、准确度、提取回收率和基质效应均符合生物样品定量分析的要求。LWK-126在人、大鼠、比格犬肝微粒体中
孵育60 min后的剩余百分率分别为（33.17±4.52）％、（3.14±6.73）％、（1.38±5.85）％，半衰期分别为33.15、11.76、5.62 min，清除
率分别为38.45、118.81、245.76 μL/（min·mg）。结论：成功建立了肝微粒体中LWK-126质量浓度的测定方法。LWK-126在不同种
属肝微粒体中的代谢稳定性排序依次为人＞大鼠＞比格犬。
关键词 LWK-126；超高效液相色谱-串联质谱法；肝微粒体；代谢稳定性

Study on Metabolic Stability of Foretinib Derivative LWK-126 in Liver Microsomes of Different Species
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WANG Zhongyuan ，YANG Huali ，CUI Xing ，ZHU Gaofeng ，TANG Lei ，LIAO Weike（1. Dept. of Pharmacy，
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Guizhou Provincial People’s Hospital，Guiyang 550004，China；2. Guizhou Provincial Engineering Technology
Research Center for Chemical Drug R&D，Guiyang 550004，China）
ABSTRACT OBJECTIVE：To establish the method for the concentration determination of foretinib derivative LWK-126 in liver
microsomes，and to study its metabolism stability in liver microsomes of rats，Beagle dogs and human. METHODS：In the in vitro
incubation system of liver microsomes，LWK-126 was dissolved in liver microsomal incubation systems of rats，Beagle dog and
human initiated by reduced nicotinamide adenine dinucleotide phosphate solution. After incubation in water at 37 ℃ for 0，5，10，20，
30 and 60 min，the reaction was terminated with acetonitrile containing internal standard（1 μg/mL tolbutamide）. UPLC-MS/MS
method was applied to determine the concentration of LWK-126 in the incubation systems. The determination was performed on
Waters BEH C18 column with mobile phase consisted of water（containing 0.1％ formic acid）-acetonitrile（containing 0.1％ formic
acid）by gradient elution at the flow rate of 0.4 mL/min. The column temperature was 30 ℃，and the sample size was 2 μL. The
mass spectral analysis was performed in a positive electrospray ionization mode，and the full MS experiment was run with the
selective reaction monitoring mode with a scanning range of m/z 50→1 200. Taking the concentration of LWK-126 at 0 min as
reference，the remaining percentage and the enzyme kinetic parameters were calculated. RESULTS：The linear range of LWK-126
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was 0.05-15 μg/mL（R ＝0.999 2）；the lower limit of quantification was 0.05 μg/mL，and the lowest detection limit was 0.01
μg/mL. The precision，accuracy，extraction recovery and matrix effect all met the analysis requirements of biological samples. The
remaining percentage of LWK-126 in liver microsomes of human，rats and Beagle dogs for 60 min were（33.17±4.52）％，（3.14±
6.73）％，（1.38±5.85）％；t 1/2 of them were 33.15，11.76，5.62 min；the clearance rates were 38.45，118.81，245.76 μL/（min·mg），
respectively. CONCLUSIONS： The method for the content
Δ 基金项目：国家自然科学基金青年科学基金项目（No.81903472）； determination of LWK-126 in liver microsomes is established
贵州省高层次创新型人才-百层次人才项目（No.黔科合平台人才
successfully. The order of metabolic stability of LWK-126 in
〔2016〕4015） liver microsomes of different species is human＞rats＞Beagle
＊主管药师，硕士。研究方向：药物分析、医院药学。E-mail：
dogs.
154366202@qq.com
# 通信作者：副教授，博士。研究方向：创新药物。电话：0851- KEYWORDS LWK-126；UPLC-MS/MS；Liver microsomes；
86908318。E-mail：641212891@qq.com Metabolic stability

中国药房 2021年第32卷第11期 China Pharmacy 2021 Vol. 32 No. 11 ·1325 ·