干旱区生态环境知识资源中心(Arid): Development and Validation of a Sensitive and Specific LC-MS/MS Method for IWR-1-Endo, a Wnt Signaling Inhibitor: Application to a Cerebral Microdialysis Study

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DOI	10.3390/molecules27175448
	Development and Validation of a Sensitive and Specific LC-MS/MS Method for IWR-1-Endo, a Wnt Signaling Inhibitor: Application to a Cerebral Microdialysis Study
	Nair, Sreenath; Davis, Abigail; Campagne, Olivia; Schuetz, John D.; Stewart, Clinton F.
通讯作者	Stewart, CF
来源期刊	MOLECULES
EISSN	1420-3049
出版年	2022
卷号	27 期号:17
英文摘要	IWR-1-endo, a small molecule that potently inhibits the Wnt/beta-catenin signaling pathway by stabilizing the AXIN2 destruction complex, can inhibit drug efflux at the blood-brain barrier. To conduct murine cerebral microdialysis research, validated, sensitive, and reliable liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods were used to determine IWR-1-endo concentration in the murine plasma and brain microdialysate. IWR-1-endo and the internal standard (ISTD) dabrafenib were extracted from murine plasma and microdialysate samples by a simple solid-phase extraction protocol performed on an Oasis HLB mu Elution plate. Chromatographic separation was executed on a Kinetex C-18 (100A, 50 x 2.1 mm, 4 mu m particle size) column with a binary gradient of water and acetonitrile, each having 0.1% formic acid, pumped at a flow rate of 0.6 mL/min. Detection by mass spectrometry was conducted in the positive selected reaction monitoring ion mode by monitoring mass transitions 410.40 > 344.10 (IWR-1-endo) and 520.40 > 307.20 (ISTD). The validated curve range of IWR-1-endo was 5-1000 ng/mL for the murine plasma method (r(2) >= 0.99) and 0.5-500 ng/mL for the microdialysate method (r(2) >= 0.99). The lower limit of quantification (LLOQ) was 5 ng/mL and 0.5 ng/mL for the murine plasma and microdialysate sample analysis method, respectively. Negligible matrix effects were observed in murine plasma and microdialysate samples. IWR-1-endo was extremely unstable in murine plasma. To improve the stability of IWR-1-endo, pH adjustments of 1.5 were introduced to murine plasma and microdialysate samples before sample storage and processing. With pH adjustment of 1.5 to the murine plasma and microdialysate samples, IWR-1-endo was stable across several tested conditions such as benchtop, autosampler, freeze-thaw, and long term at -80 degrees C. The LC-MS/MS methods were successfully applied to a murine pharmacokinetic and cerebral microdialysis study to characterize the unbound IWR-1-endo exposure in brain extracellular fluid and plasma.
英文关键词	IWR-1-endo Wnt signaling inhibitor LC-MS MS solid-phase extraction pharmacokinetics cerebral microdialysis bioanalysis
类型	Article
语种	英语
开放获取类型	gold, Green Published
收录类别	SCI-E
WOS记录号	WOS:000851775900001
WOS关键词	PLASMA ; MEDULLOBLASTOMA ; EXPRESSION ; MOLECULES
WOS类目	Biochemistry & Molecular Biology ; Chemistry, Multidisciplinary
WOS研究方向	Biochemistry & Molecular Biology ; Chemistry
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/393818
推荐引用方式 GB/T 7714	Nair, Sreenath,Davis, Abigail,Campagne, Olivia,et al. Development and Validation of a Sensitive and Specific LC-MS/MS Method for IWR-1-Endo, a Wnt Signaling Inhibitor: Application to a Cerebral Microdialysis Study[J],2022,27(17).
APA	Nair, Sreenath,Davis, Abigail,Campagne, Olivia,Schuetz, John D.,&Stewart, Clinton F..(2022).Development and Validation of a Sensitive and Specific LC-MS/MS Method for IWR-1-Endo, a Wnt Signaling Inhibitor: Application to a Cerebral Microdialysis Study.MOLECULES,27(17).
MLA	Nair, Sreenath,et al."Development and Validation of a Sensitive and Specific LC-MS/MS Method for IWR-1-Endo, a Wnt Signaling Inhibitor: Application to a Cerebral Microdialysis Study".MOLECULES 27.17(2022).