Development of a high-performance liquid chromatographic-mass spectrometric method for the determination of cellular levels of the tyrosine kinase inhibitors lapatinib and dasatinib

被引:36
作者
Roche, Sandra [1 ]
McMahon, Gillian [2 ]
Clynes, Martin [1 ]
O'Connor, Robert [1 ]
机构
[1] Dublin City Univ, Natl Inst Cellular Biotechnol, Dublin 9, Ireland
[2] Dublin City Univ, Sch Chem Sci, Dublin 9, Ireland
来源
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 2009年 / 877卷 / 31期
基金
爱尔兰科学基金会;
关键词
Lapatinib; Dasatinib; Liquid-liquid extraction; Liquid chromatography mass spectrometry; Tyrosine kinase inhibitor; QUANTITATIVE BIOANALYSIS; ANTITUMOR-ACTIVITY; CANCER-THERAPY; MOUSE PLASMA; IN-VITRO; RESISTANCE; IMATINIB; GW572016; TRANSPORTERS; VALIDATION;
D O I
10.1016/j.jchromb.2009.10.008
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A highly sensitive and selective liquid chromatography tandem mass spectrometry (LC-MS/MS) method has been developed to quantify cellular levels of the tyrosine kinase inhibitors (TKIs) dasatinib (Sprycel (TM)) and lapatinib (Tykerb (TM), Tyverb (TM)). Cellular samples were extracted with a tert-butyl methyl ether: acetonitrile (3: 1, v/v): 1 M ammonium formate pH 3.5 (8: 1, v/v) mixture. Separation was achieved on a Hyperclone BDS C18 (150 mm x 2.0 mm 3 mu m) column with isocratic elution using a mobile phase of acetonitirile-10 mM ammonium formate. pH 4 (54:46, v/v), at a flow rate of 0.2 mL/min. The TKIs were quantified using a triple quadrupole mass spectrometer which was operated in multi-reaction-monitoring mode employing positive electrospray ionisation. The limit of detection and limit of quantification for lapatinib was determined to be 15 and 31 pg on column, respectively. The limit of detection and quantification for dasatinib was 3 and 15 pg on column, respectively. The method allowed for sensitive and accurate determination of cellular levels of dasatinib and lapatinib. in addition, we examined the potential for this method to be utilised to quantitate other TKIs, using gefitinib, erlotinib, imatinib and sorafenib as examples. In principle, these agents were also quantifiable by this method, however, no drug specific validation studies were undertaken with these TKIs. The data indicates that in the cancer cell-line model, DLKP, significantly more lapatinib accumulates in cells in comparison to dasatinib. Additionally, over-expression of the membrane protein drug transporter, P-glycoprotein (P-gp) a common cancer drug resistance mechanism, greatly reduces the cellular accumulation of dasatinib but not of lapatinib. (C) 2009 Elsevier B.V. All rights reserved.
引用
收藏
页码:3982 / 3990
页数:9
相关论文
共 29 条
[11]   Hydrophilic interaction liquid chromatography/tandem mass spectrometry for the simultaneous determination of dasatinib, imatinib and nilotinib in mouse plasma [J].
Hsieh, Yunsheng ;
Galviz, Gerica ;
Zhou, Qiao ;
Duncan, Christine .
RAPID COMMUNICATIONS IN MASS SPECTROMETRY, 2009, 23 (09) :1364-1370
[12]   Protein tyrosine kinase structure and function [J].
Hubbard, SR ;
Till, JH .
ANNUAL REVIEW OF BIOCHEMISTRY, 2000, 69 :373-398
[13]  
Jemal M, 2000, BIOMED CHROMATOGR, V14, P422, DOI 10.1002/1099-0801(200010)14:6<422::AID-BMC25>3.0.CO
[14]  
2-I
[15]   Preclinical pharmacokinetics and in vitro metabolism of dasatinib (BMS-354825): a potent oral multi-targeted kinase inhibitor against SRC and BCR-ABL [J].
Kamath, Amrita V. ;
Wang, Jian ;
Lee, Francis Y. ;
Marathe, Punit H. .
CANCER CHEMOTHERAPY AND PHARMACOLOGY, 2008, 61 (03) :365-376
[16]   Tyrosine kinases as targets for cancer therapy [J].
Krause, DS ;
Van Etten, RA .
NEW ENGLAND JOURNAL OF MEDICINE, 2005, 353 (02) :172-187
[17]   Discovery of N-(2-chloro-6-methylphenyl)-2-(6-(4-(2-hydroxyethyl)-piperazin-1-yl)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide (BMS-354825), a dual Src/Abl kinase inhibitor with potent antitumor activity in preclinical assays [J].
Lombardo, LJ ;
Lee, FY ;
Chen, P ;
Norris, D ;
Barrish, JC ;
Behnia, K ;
Castaneda, S ;
Cornelius, LAM ;
Das, J ;
Doweyko, AM ;
Fairchild, C ;
Hunt, JT ;
Inigo, I ;
Johnston, K ;
Kamath, A ;
Kan, D ;
Klei, H ;
Marathe, P ;
Pang, SH ;
Peterson, R ;
Pitt, S ;
Schieven, GL ;
Schmidt, RJ ;
Tokarski, J ;
Wen, ML ;
Wityak, J ;
Borzilleri, RM .
JOURNAL OF MEDICINAL CHEMISTRY, 2004, 47 (27) :6658-6661
[18]   Human lung carcinoma cell line DLKP contains 3 distinct subpopulations with different growth and attachment properties [J].
McBride, S ;
Meleady, P ;
Baird, A ;
Dinsdale, D ;
Clynes, M .
TUMOR BIOLOGY, 1998, 19 (02) :88-103
[19]   Drug resistance in cancer - searching for mechanisms, markers and therapeutic agents [J].
O'Connor, Robert ;
Clynes, Martin ;
Dowling, Paul ;
O'Donovan, Norma ;
O'Driscoll, Lorraine .
EXPERT OPINION ON DRUG METABOLISM & TOXICOLOGY, 2007, 3 (06) :805-817
[20]  
O'Connor R, 2007, ANTICANCER RES, V27, P1267