Abstract
A rapid and sensitive HPLC–MS/MS assay was established and validated to quantify LXK4, a novel potent B-RafV600E inhibitor in dog plasma. The samples prepared by liquid–liquid extraction using ethyl acetate were separated on a Phenomenex C8 column with mobile phase of water–methanol. LXK4 and IS were all detected by monitoring the precursor → product ion transition of m/z 413.4 → 345.4 and 401.3 → 333.3, respectively, using multiple reaction monitoring (MRM) scan mode with positive ionization mode. The liner calibration curves for LXK4 in dog plasma ranged from 0.5 to 1000 ng mL−1 with the lower limit of quantification of 0.5 ng mL−1. The intra- and inter-day precisions were lower than 12.0%, and the accuracy ranged from −4.6 to 9.5%. The validated assay was successfully used as a new approach to study the pharmacokinetics of LXK4 in beagle dogs and the oral bioavailability of LXK4 was determined to be 30.5%.
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Heath E, Kaufman K, Christopherson R (2011) Int J Biochem Cell Biol 43:29–32
Dhillon AS, Hagan S, Rath O, Kolch W (2007) Oncogene 26:3279–3290
Marais R, Marshall CJ (1996) Cancer Surv 27:101–125
Peyssonnaux C, Eychène A (2001) Biol Cell 93:53–62
Nazarian R, Shi H, Wang Q, Kong X, Koya RC, Lee H, Chen Z, Lee M-K, Attar N, Sazegar H (2010) Nature 468:973–977
Yuen ST, Davies H, Chan TL, Ho JW, Bignell GR, Cox C, Stephens P, Edkins S, Tsui WW, Chan AS (2002) Cancer Res 62:6451–6455
Kimura ET, Nikiforova MN, Zhu Z, Knauf JA, Nikiforov YE, Fagin JA (2003) Cancer Res 63:1454–1457
Dhomen N, Marais R (2007) Curr Opin Genet Dev 17:31–39
Karasarides M, Chiloeches A, Hayward R, Niculescu-Duvaz D, Scanlon I, Friedlos F, Ogilvie L, Hedley D, Martin J, Marshall CJ (2004) Oncogene 23:6292–6298
Holderfield M, Deuker MM, McCormick F, McMahon M (2014) Nat Rev Cancer 14:455–467
Davies H, Bignell GR, Cox C, Stephens P, Edkins S, Clegg S, Teague J, Woffendin H, Garnett MJ, Bottomley W (2002) Nature 417:949–954
Fiskus W, Mitsiades N (2016) Annu Rev Med 67:29–43
Wang X, Kim J (2012) J Med Chem 55:7332–7341
Wilhelm SM, Carter C, Tang L, Wilkie D, McNabola A, Rong H, Chen C, Zhang X, Vincent P, McHugh M (2004) Cancer Res 64:7099–7109
Eisen T, Ahmad T, Flaherty K, Gore M, Kaye S, Marais R, Gibbens I, Hackett S, James M, Schuchter L (2006) Br J Cancer 95:581–586
Yang H, Higgins B, Kolinsky K, Packman K, Go Z, Iyer R, Kolis S, Zhao S, Lee R, Grippo JF (2010) Cancer Res 70:5518–5527
Falchook G, Long G, Kurzrock R, Kim K, Arkenau H, Brown M, Hamid O, Infante J, Millward M, Pavlick A (2012) Lancet 379:1893–1901
Rheault TR, Stellwagen JC, Adjabeng GM, Hornberger KR, Petrov KG, Waterson AG, Dickerson SH, Mook RA Jr, Laquerre SG, King AJ (2013) ACS Med Chem Lett 4:358–362
Bollag G, Tsai J, Zhang J, Zhang C, Ibrahim P, Nolop K, Hirth P (2012) Nat Rev Drug Discov 11:873–886
Chapman PB, Hauschild A, Robert C, Haanen JB, Ascierto P, Larkin J, Dummer R, Garbe C, Testori A, Maio M (2011) N Engl J Med 364:2507–2516
Young K, Minchom A, Larkin J (2012) Future Oncol 8:499–507
Hauschild A, Grob J-J, Demidov LV, Jouary T, Gutzmer R, Millward M, Rutkowski P, Blank CU, Miller WH, Kaempgen E (2012) Lancet 380:358–365
Chu EY, Wanat KA, Miller CJ, Amaravadi RK, Fecher LA, Brose MS, McGettigan S, Giles LR, Schuchter LM, Seykora JT (2012) J Am Acad Dermatol 67:1265–1272
Anforth R, Fernandez-Peñas P, Long GV (2013) Lancet Oncol 14:e11–e18
Li X, Shen J, Tan L, Zhang Z, Gao D, Luo J, Cheng H, Zhou X, Ma J, Ding K (2016) Bioorg Med Chem Lett 26:2760–2763
FDA (2001) Guidance for industry: bioanalytical methods validation, 2001. http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/UCM070107.pdf. Accessed May 2001
Acknowledgements
This study was partially supported by the National Natural Science Foundation of China (#21302186); the Guangdong Province Natural Science Foundation (#2014A030313735); the Guangdong Science and Technology Planning Founding (#2013A022100019); and Zhongshan Introducing Talent Award.
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E. Jiang and J. Deng contributed equally to the work.
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Jiang, E., Deng, J., Guo, J. et al. Determination of A Novel Selective B-RafV600E Inhibitor (LXK4) in Dog Plasma by HPLC–MS/MS and its Application in a Pharmacokinetic Study. Chromatographia 80, 71–76 (2017). https://doi.org/10.1007/s10337-016-3195-9
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DOI: https://doi.org/10.1007/s10337-016-3195-9