Development of Fast and Simple LC-ESI-MS/MS Technique for the Quantification of Regorafenib; Application to Pharmacokinetics in Healthy Rabbits

Yatha       Ravi; Darna      Bhikshapathi; Shankar       Cheruku; Bigala       Rajkamal

doi:10.2174/1573412916666191111144707

Abstract

Background: A simple quantification technique by liquid chromatography electrospray ionization- tandem mass spectrometry (LC-ESI-MS/MS) is required for regorafenib in biological matrices with bioavailability studies in healthy rabbits, when compared with reported techniques.

Objective: The main aim of the research work was to develop a validated LC-ESI-MS/MS technique for the quantification of regorafenib and application to bioavailability studies in healthy rabbits.

Methods: Chromatographic separation was achieved with hypersil-C₁₈ analytical column (50mm×4.6 mm, 4μm) and mobile phase composition of acetonitrile and 5mM ammonium acetate in the proportion of 70:30. The mobile phase was infused into the column with high pressure to get a 0.7 ml/min flow rate. The total retention time of the analyte is promising when compared with the existed methods for regorafenib. Quantitation was processed by monitoring transitions of m/z -483.0/262.0 and 450.0/260.0 for regorafenib and internal standard respectively in multiple reaction monitoring.

Results: The linearity equation and correlation coefficient (R2) findings were y =0.9948x+2.6624 and 0.998 respectively. The intra and inter-day precision of the developed technique was found between 1.00 – 8.50% for the QC-samples (2, 4, 240 and 480ng/ml). From bioavailability study, the drug was shown T_max of 3.688 ± 0.754; average AUC_0→α and AUC_0→t were 6476.81 ± 259.59 and 6213.845 ± 257.892 respectively and C_max was found to be 676.91 ± 22.045 in healthy rabbits.

Conclusion: The developed technique was validated and successfully applied in the pharmacokinetic study of the drug (40 mg tablet) administered through the oral route in healthy rabbits.

Keywords: Regorafenib, tyrosine kinase inhibitor, bioavailability studies, LC-MS/MS, matrix effect, metabolites.

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[1] 

Bayer announces New data on oncology portfolio to be presented at the ECCO-ESMO congress., 2009 2009-09-19;, 
[2] 
Zhang, G.; Kodani, S.; Hammock, B.D. Stabilized epoxygenated fatty acids regulate inflammation, pain, angiogenesis and cancer. Prog. Lipid Res.,  2014, 53, 108-123.
[http://dx.doi.org/10.1016/j.plipres.2013.11.003] [PMID:  24345640] 
[3] 
Hwang, S.H.; Wecksler, A.T.; Zhang, G.; Morisseau, C.; Nguyen, L.V.; Fu, S.H.; Hammock, B.D. Synthesis and biological evaluation of sorafenib- and regorafenib-like sEH inhibitors. Bioorg. Med. Chem. Lett.,  2013, 23(13), 3732-3737.
[http://dx.doi.org/10.1016/j.bmcl.2013.05.011] [PMID:  23726028] 
[4] 
Kim, J.; Ulu, A.; Wan, D.; Yang, J.; Hammock, B.D.; Weiss, R.H. Addition of DHA synergistically enhances the efficacy of reforafenib for kidney cancer therapy. Mol. Cancer Ther.,  2016, 15(5), 890-898.
[http://dx.doi.org/10.1158/1535-7163.MCT-15-0847] [PMID:  26921392] 
[5] 
Kim, J.; Ulu, A.; Wan, D.; Yang, J.; Hammock, B.D.; Weiss, R.H. Addition of dha synergistically enhances the efficacy of regorafenib for kidney cancer therapy. Mol. Cancer Ther.,  2016, 15(5), 890-898.
[http://dx.doi.org/10.1158/1535-7163.MCT-15-0847] [PMID:  26921392] 
[6] 
Hotta, K.; Ueyama, J.; Tatsumi, Y.; Tsukiyama, I.; Sugiura, Y.; Saito, H.; Matsuura, K.; Hasegawa, T. Lack of Contribution of multidrug resistance-associated protein and organic anion-transporting polypeptide to pharmacokinetics of regorafenib, a novel multi-kinase inhibitor, in rats. Anticancer Res.,  2015, 35(9), 4681-4689.
[PMID:  26254357] 
[7] 
Allard, M.; Khoudour, N.; Rousseau, B.; Joly, C.; Costentin, C.; Blanchet, B.; Tournigand, C.; Hulin, A. Simultaneous analysis of regorafenib and sorafenib and three of their metabolites in human plasma using LC-MS/MS. J. Pharm. Biomed. Anal.,  2017, 142(142), 42-48.
[http://dx.doi.org/10.1016/j.jpba.2017.04.053] [PMID:  28494338] 
[8] 
Kazuma, F. Masatomo, Miura.; shibata, Hiroyuki. Quantitative determination of regorafenib and its two major metabolites in human plasma with high-performance liquid chromatography and ultraviolet detection. Biomed. Chromatogr.,  2016, 30(10), 112-119.
[9] 
Mevlut, A. Development and validation of a new spectrophotometric method for the determination of regorafenib in pure and tablet dosage form. Lat. Am. J. Pharm.,  2018, 37(7), 1349-1353.
[10] 
Weiping, J.I.; Qingwei, Z.H.A.N.G.; Lufeng, H.U. Development of a Simple LC-MS Assay for determination of regorafenib in rat plasma and its application to a pharmacokinetic study. Lat. Am. J. Pharm.,  2014, 33(4), 607-612.
[11] 
Hafner, F.T.; Werner, D.; Kaiser, M. Determination of regorafenib (BAY 73-4506) and its major human metabolites BAY 75-7495 (M-2) and BAY 81-8752 (M-5) in human plasma by stable-isotope dilution liquid chromatography-tandem mass spectrometry. Bioanalysis,  2014, 6(14), 1923-1937.
[http://dx.doi.org/10.4155/bio.14.52] [PMID:  25158964] 
[12] 
Dino, Luethi; Selvi, Durmus Alfred, H.; Schinkel, Jan, H. M.;
	Schellens, Jos, H.; Beijnen, Rolf, W. Liquid chromatography–
	tandem mass spectrometric assay for the multikinase inhibitor
	regorafenib in plasma. SparidansBiomedical chromatography.,
	2014, 28(10), 1366-70.. 
[13] 
Gudisa, K. A validated RP-HPLC method for estimation of regorafenib in bulk and tablet dosage form. Experiment,  2012, 3(1), 174-181.
[14] 
Ramesh, Jayaprakash Senthil, Kumar Natesan. A validated stability
	indicating rp-hplc method for the estimation of an anti-cancer
	drug regorafenib in pure and pharmaceutical dosage form. J Pharm
	chem., 2017, 4, 81-9., 
[15] 
Fujita, K.; Miura, M.; Shibata, H. Quantitative determination of regorafenib and its two major metabolites in human plasma with high-performance liquid chromatography and ultraviolet detection. Biomed. Chromatogr.,  2016, 30(10), 1611-1617.
[http://dx.doi.org/10.1002/bmc.3730] [PMID:  27037679] 
[16] 
Jithendra, D. Pratap, Kumar Patra.; Suresh, P. RP-HPLC method development and validation of Regorafenib in pure form and pharmaceutical dosage form. Asian J. Res. Chem,  2018, 11(4), 763-767.
[http://dx.doi.org/10.5958/0974-4150.2018.00134.7] 
[17] 
Kumar, S. Moola, BalaSekhara Reddy Challa.; Chandrasekhar, KothapalliBannoth. Quantification of tolvaptan in rabbit plasma by LC–MS/MS: Application to a pharmacokinetic study. J. Pharm. Anal.,  2015, 5, 371-377.
[http://dx.doi.org/10.1016/j.jpha.2014.09.001] 
[18] 
Tejas, D. Komal, Chaudhary.; Priti, Mehta. Development of LC-MS/MS method for determination of iloperidone in rabbit plasma: application to a pharmacokinetic study. Int. J. Pharm. Pharm. Sci.,  2013, 7(4), 294-297.
[19] 
Kumar Moola, S. BalaSekhara Reddy, Challa; Chandrasekhar, K.B. Bioanalytical method development and validation of eszopiclone in rabbit plasma by HPLC-MS/MS and its application to pharmacokinetic study. International Journal of Biological & Pharmaceutical Research,  2014, 5(9), 719-727.
[20] 
Neelima, P. Chandanam, Sreedhar.; SreenivasRao, T.; Veera Reddy, S. Development and validation of liquid chromatography- tandem mass pectrometry for determination of olanzapine in rabbit plasma. DHR International Journal of Pharmaceutical Sciences,  2014, 5(1), 151-159.
[21] 
Chandrasekar, M.J.N.; Chandrasekar, A.J.; Krishnaraj, K.; Muralidharan, S.; Rajan, S.; Suresh, B. Liquid chromatography-mass spectrometry determination of cetrizine hydrochloride in rabbit plasma. Asian J. Chem.,  2009, 21(8), 5821-5828.
[22] 

FDA Guidance for Industry, Bioanalytical Method Validation, US
	Department of Health and Human Services, Food and Drug Administration,
	Centre for Drug Evaluation and Research (CDER),
	Center for Veterinary Medicine (CVM) May, , 2001.
[23] 

Guideline on bioanalytical method validation,, 2011.
[24] 

 Harmonized Tripartite Guideline, Validation of Analytical Procedure:
	Methodology, IFPMA Proceedings of the International Conference
	on Harmonization, Geneva, 1996.
[25] 
del Mar Ramírez Fernández, M.; Wille, S.M.R.; Samyn, N. Quantitative method validation for the analysis of 27 antidepressants and metabolites in plasma with ultraperformance liquid chromatography-tandem mass spectrometry. Ther. Drug Monit.,  2012, 34(1), 11-24.
[http://dx.doi.org/10.1097/FTD.0b013e31823bf0fd] [PMID:  22210095] 
[26] 
Woźniakiewicz, M.; Wietecha-Posłuszny, R.; Moos, A.; Wieczorek, M.; Knihnicki, P.; Kościelniak, P. Development of microextraction by packed sorbent for toxicological analysis of tricyclic antidepressant drugs in human oral fluid. J. Chromatogr. A,  2014, 1337, 9-16.
[http://dx.doi.org/10.1016/j.chroma.2014.02.037PMID: 24636563] 
[27] 
Chambers, E.E.; Woodcock, M.J.; Wheaton, J.P.; Pekol, T.M.; Diehl, D.M. Systematic development of an UPLC-MS/MS method for the determination of tricyclic antidepressants in human urine. J. Pharm. Biomed. Anal.,  2014, 88, 660-665.
[http://dx.doi.org/10.1016/j.jpba.2013.09.001] [PMID:  24239905] 
[28] 
Patel, D.S.; Sharma, N.; Patel, M.C.; Patel, B.N.; Shrivastav, P.S.; Sanyal, M. Development and validation of a selective and sensitive LC-MS/MS method for determination of cycloserine in human plasma: application to bioequivalence study. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci.,  2011, 879(23), 2265-2273.
[http://dx.doi.org/10.1016/j.jchromb.2011.06.011] [PMID:  21727043] 
[29] 
Yadav, M.; Shrivastav, P.S. Incurred sample reanalysis (ISR): a decisive tool in bioanalytical research. Bioanalysis,  2011, 3(9), 1007-1024.
[http://dx.doi.org/10.4155/bio.11.76] [PMID:  21545349] 
[30] 
Hewavitharana, A.K. Matrix matching in liquid chromatography-mass spectrometry with stable isotope labelled internal standards--is it necessary? J. Chromatogr. A,  2011, 1218(2), 359-361.
[http://dx.doi.org/10.1016/j.chroma.2010.11.047] [PMID:  21159347] 
[31] 
Bonfiglio, R.; King, R.C.; Olah, T.V.; Merkle, K. The effects of sample preparation methods on the variability of the electrospray ionization response for model drug compounds. Rapid Commun. Mass Spectrom.,  1999, 13(12), 1175-1185.
[http://dx.doi.org/10.1002/(SICI)1097-0231(19990630)13:12<1175:AID-RCM639>3.0.CO;2-0] [PMID:  10407294] 
[32] 
Cheng, Y.; Lu, Z.; Neue, U. Ultrafast liquid chromatography/ultraviolet and liquid chromatography/tandem mass spectrometric analysis. Rapid Commun. Mass Spectrom.,  2001, 15(2), 141-151.
[http://dx.doi.org/10.1002/1097-0231(20010130)15:2<141:AID-RCM201>3.0.CO;2-I] [PMID:  11180543] 
[33] 
Hayward, M.J.; Munson, J.L.; Conneely, G.; Hargiss, L.O. What Limits Productivity in High Flow Fast LC/MS? Proceedings of the 49th ASMS Conference on Mass Spectrometry and Allied Topics, 2001.Chicago, IL
[34] 
Buhrman, D.L.; Price, P.I.; Rudewiczcor, P.J. Quantitation of SR 27417 in human plasma using electrospray liquid chromatography-tandem mass spectrometry: A study of ion suppression. J. Am. Soc. Mass Spectrom.,  1996, 7(11), 1099-1105.
[http://dx.doi.org/10.1016/S1044-0305(96)00072-4] [PMID:  24203071] 
[35] 
Jemal, M.; Ouyang, Z. The need for chromatographic and mass resolution in liquid chromatography/tandem mass spectrometric methods used for quantitation of lactones and corresponding hydroxy acids in biological samples. Rapid Commun. Mass Spectrom.,  2000, 14(19), 1757-1765.
[http://dx.doi.org/10.1002/1097-0231(20001015)14:19<1757:AID-RCM90>3.0.CO;2-C] [PMID:  11006583] 
[36] 
FDA Guidance for Industry.Center for Drug Evaluation and Research.
	Biopharmaceutics, BioanalyticalMethod Validation. Issued 5/, . 2001.http://www.fda.gov/cder/guidance/index.htm
[37] 
Schultz, G.A.; Corso, T.N.; Prosser, S.J.; Zhang, S. A fully integrated monolithic microchip electrospray device for mass spectrometry. Anal. Chem.,  2000, 72(17), 4058-4063.
[http://dx.doi.org/10.1021/ac000325y ] [PMID:  10994965] 
[38] 
Temesi, D.; Law, B. The Effect of LC Eluent Composition on MS Responses Using Electrospray Ionization. LC GC,  1999, 17, 626-632.
[39] 
King, R.; Bonfiglio, R.; Fernandez-Metzler, C. Miller- Stein, C.; Olah, T. mechanistic investigationof ionization suppression in electrospray ionization. J. Am. Soc. Mass Spectrom.,  2000, 11, 942-950.
[http://dx.doi.org/10.1016/S1044-0305(00)00163-X] [PMID:  11073257] 
[40] 
Doerge, D.R.; Fogle, C.M.; Paule, M.G.; McCullagh, M.; Bajic, S. Analysis of methylphenidate and its metabolite ritalinicacid in monkey plasma by liquid chromatography/electrospray ionizationmass spectrometry. Rapid Commun. Mass Spectrom.,  2000, 14, 610-623.
[http://dx.doi.org/10.1002/(SICI)1097-0231(20000430)14:8<619:AID-RCM916>3.0.CO;2-2] 
[41] 
Murphy, A.T.; Kasper, S.C.; Gillespie, T.A.; Delong, A.F. Determination of xanomeline and active metabolite, N-desmethylxanomeline, in human plasma by liquid chromatography-atmospheric pressure chemical ionization mass spectrometry. J. Chromatogr. B Biomed. Appl.,  1995, 668(2), 273-280.
[http://dx.doi.org/10.1016/0378-4347(95)00080-3 ] [PMID:  7581862] 
[42] 
Tiller, P.R.; Cunniff, J.; Land, A.P.; Schwartz, J.; Jardine, I.; Wakefield, M.; Lopez, L.; Newton, J.F.; Burton, R.D.; Folk, B.M.; Buhrman, D.L.; Price, P.; Wu, D. Drug quantitation on a benchtop liquid chromatography-tandem mass spectrometry system. J. Chromatogr. A,  1997, 771(1-2), 119-125.
[http://dx.doi.org/10.1016/S0021-9673(97)00147-7 ] [PMID:  9210315] 
[43] 
Schweingruber, H.; Dunyach, J.J.; Olney, T.N.; Taylor, D.; Churchill, M.; Amad, M.; Winnik, W.; Paul, G.; Schoen, A.E.; Campbell, C. Advantages and Limitations of Increased Mass Resolution for Quantitative SRM Analysis on a Triple Stage QuadrupoleMass Spectrometer. Proceedings of the 49th ASMS Conference on Mass Spectrometry and Allied Topics, 2001.Chicago,IL
[44] 
Henion, J.; Brewer, E.; Rule, G. Sample preparation for LC/MS/MS: knowing the basic requirements andthe big picture of an lc/ms system can ensure success in most instances. Anal. Chem.,  1998, 70, 650A-656A.
[http://dx.doi.org/10.1021/ac981991q] [PMID:  9784742] 
[45] 
Simpson, H.; Berthemy, A.; Buhrman, D.; Burton, R.; Newton, J.; Kealy, M.; Wells, D.; Wu, D. High throughput liquid chromatography/mass spectrometry bioanalysis using 96-well disk solid phase extraction plate for the sample preparation. Rapid Commun. Mass Spectrom.,  1998, 12(2), 75-82.
[http://dx.doi.org/10.1002/(SICI)1097-0231(19980131)12:2<75:AID-RCM112>3.0.CO;2-C] [PMID:  9470219] 
[46] 
Watt, A.P.; Morrison, D.; Locker, K.L.; Evans, D.C. Higher throughput bioanalysis by automation of a protein precipitation assay using a 96-well format with detection by LC-MS/MS. Anal. Chem.,  2000, 72(5), 979-984.
[http://dx.doi.org/10.1021/ac9906633 ] [PMID:  10739201] 
[47] 
Allanson, J.P.; Biddlecombe, R.A.; Jones, A.E. Pleasance. The use of automated solid-phase extraction in the 96-well format for high throughput bioanalysis using liquid chromtography coupled to tandem mass spectrometry. Rapid Commun. Mass Spectrom.,  1996, 10, 811-816.
[http://dx.doi.org/10.1002/(SICI)1097-0231(199605)10:7<811:AID-RCM561>3.0.CO;2-Q] 
[48] 
Janiszewski, J.; Schneider, P.; Hoffmaster, K.; Swyden, M.; Wells, D.; Fouda, H. Automated sample preparation using membrane microtiter extraction for bioanalytical mass spectrometry. Rapid Commun. Mass Spectrom.,  1997, 11(9), 1033-1037.
[http://dx.doi.org/10.1002/(SICI)1097-0231(19970615)11:9<1033:AID-RCM951>3.0.CO;2-5] [PMID:  9204578] 
[49] 
Jemal, M.; Teitz, D.; Ouyang, Z.; Khan, S. Comparison of plasma sample purification by manual liquid-liquid extraction, automated 96-well liquid-liquid extraction and automated 96-well solid-phase extraction for analysis by high-performance liquid chromatography with tandem mass spectrometry. J. Chromatogr. B Biomed. Sci. Appl.,  1999, 732(2), 501-508.
[http://dx.doi.org/10.1016/s0378-4347(99)00323-0.]] 
[50] 
Jemal, M.; Teitz, D.; Ouyang, Z.; Khan, S. Comparison of plasma sample purification by manual liquid-liquid extraction, automated 96-well liquid-liquid extraction and automated 96-well solid-phase extraction for analysis by high-performance liquid chromatography with tandem mass spectrometry. J. Chromatogr. B Biomed. Sci. Appl.,  1999, 732(2), 501-508.
[http://dx.doi.org/10.1016/S0378-4347(99)00323-0] [PMID:  10517373] 
[51] 
Zhang, N.; Hoffman, K.L.; Li, W.; Rossi, D.T. Semi-automated 96-well liquid-liquid extraction for quantitation of drugs in biological fluids. J. Pharm. Biomed. Anal.,  2000, 22(1), 131-138.
[http://dx.doi.org/10.1016/S0731-7085(99)00247-2] [PMID:  10727132] 
[52] 
Murphy, A.T.; Johnson, J.T.; Ackermann, B.L.; Gillespie, T.A.; Garner, C.O. Bioavailability of zyprexa lead generation formulations in dog plasma using LC/MS/MS Detection of protein precipitation supernatants injected From 96-Well microtiterplates. Proceedings of the 46th ASMS Conference on Mass Spectrometry and Allied Topics, 1998.Orlando, FL
[53] 
Sanson, A.L.; Silva, S.C.R.; Martins, M.C.G.; Giusti-Paiva, A.; Maia, P.P.; Martins, I. Liquid-liquid extraction combined with high performance liquid chromatography-diode arrayultra-violet for simultaneous determination of antineoplastic drugs inplasma. Braz. J. Pharm. Sci.,  2011, 47, 363-371.
[http://dx.doi.org/10.1590/S1984-82502011000200017] 
[54] 
Ahmad, S.; Kalra, H.; Gupta, A.; Raut, B.; Hussain, A.; Rahman, M.A. HybridSPE: A novel technique to reduce phospholipid-based matrix effect in LC-ESI-MS Bioanalysis. J. Pharm. Bioallied Sci.,  2012, 4(4), 267-275.
[http://dx.doi.org/10.4103/0975-7406.168023] [PMID:  23248558] 
[55] 
Zhou, J.; Gao, S.; Zhang, F.; Jiang, B.; Zhan, Q.; Cai, F.; Li, J.; Chen, W. Liquid chromatography-tandem mass spectrometry method for simultaneous determination of seven commonly used anticancer drugs in human plasma. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci.,  2012, 906, 1-8.
[http://dx.doi.org/10.1016/j.jchromb.2012.07.033] [PMID:  22959676] 
[56] 
Liu, G.; Aubry, A.F. Best Practices.Biological Sample Preparation
	For LC-MS Bioanalysis. In: Handbook of LC-MS Bioanalysis.,
	2013, 165-184.
[57] 
Jessome, L.L.; Volmer, D.A. Ion Suppression: A Major Concern for Mass Spectrometry. LC GC N. Am.,  2006, 24, 498-510.
[58] 
Buszewski, B.; Noga, S. Hydrophilic interaction liquid chromatography (HILIC)--a powerful separation technique. Anal. Bioanal. Chem.,  2012, 402(1), 231-247.
[http://dx.doi.org/10.1007/s00216-011-5308-5] [PMID:  21879300] 
[59] 
Bowers, L.D. Analytical goals in therapeutic drug monitoring. Clin. Chem.,  1998, 44(2), 375-380.
[PMID:  9474048] 
[60] 

U.S. Department of Health and Human Services Food and Drug
	Administration, Guidance for Industry: Bioanalytical Method Validation,
	2001, 1-22., 

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Development of Fast and Simple LC-ESI-MS/MS Technique for the Quantification of Regorafenib; Application to Pharmacokinetics in Healthy Rabbits

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