To develop a reproducible microdialysis-tumor homogenate method for the study of the intratumor distribution of a highly hydrophobic anticancer drug (SN-38; 7-ethyl-10-hydroxycamptothecin) in neuroblastoma patient-derived xenografts.
We studied the nonspecific binding of SN-38 to the microdialysis tubing in the presence of 2-hydroxypropyl-beta-cyclodextrin (HPBCD) in the perfusate. We calibrated the microdialysis probes by the zero flow rate (ZFR) method and calculated the enhancement factor (f = extrapolated SN-38 concentration at the ZFR / SN-38 concentration in the dialysed solution) of HPBCD. We characterized the extravasation of HPBCD to tumors engrafted in mice. In vivo microdialysis and terminal homogenate data at the steady state (subcutaneous pump infusions) were used to calculate the volume of distribution of unbound SN-38 (Vu,tumor) in neuroblastoma.
HPBCD (10% w/v) in the perfusate prevented the nonspecific binding of SN-38 to the microdialysis probe and enhanced SN-38 recovery (f = 1.86). The extravasation of HPBCD in the tumor during microdialysis was lower than 1%. Vu,tumor values were above 3 mL/g tumor for both neuroblastoma models and suggested efficient cellular penetration of SN-38.
The method contributes to overcome the limitations of the microdialysis technique in hydrophobic drugs and provides a powerful tool to characterize compartmental anticancer drug distribution in xenografts.
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- f :
Fluorinated ethylene propylene
- SN-38 C:
- SN-38 L:
Tumor extracellular fluid
Zero flow rate
Patel KJ, Tredan O, Tannock IF. Distribution of the anticancer drugs doxorubicin, mitoxantrone and topotecan in tumors and normal tissues. Cancer Chemother Pharmacol. 2013;72(1):127–38.
Minchinton AI, Tannock IF. Drug penetration in solid tumours. Nat Rev Cancer. 2006;6(8):583–92.
Kratz F, Warnecke A. Finding the optimal balance: challenges of improving conventional cancer chemotherapy using suitable combinations with nano-sized drug delivery systems. J Control Release. 2012;164(2):221–35.
Provenzano PP, Cuevas C, Chang AE, Goel VK, Von Hoff DD, Hingorani SR. Enzymatic targeting of the stroma ablates physical barriers to treatment of pancreatic ductal adenocarcinoma. Cancer Cell. 2012;21(3):418–29.
Fletcher JI, Haber M, Henderson MJ, Norris MD. ABC transporters in cancer: more than just drug efflux pumps. Nat Rev Cancer. 2010;10(2):147–56.
Holback H, Yeo Y. Intratumoral drug delivery with nanoparticulate carriers. Pharm Res. 2011;28(8):1819–30.
Tuntland T, Ethell B, Kosaka T, Blasco F, Zang RX, Jain M, et al. Implementation of pharmacokinetic and pharmacodynamic strategies in early research phases of drug discovery and development at Novartis Institute of Biomedical Research. Front Pharmacol. 2014;5:174.
Muller M. Microdialysis in clinical drug delivery studies. Adv Drug Deliv Rev. 2000;45(2–3):255–69.
Wang Y, Welty DF. The simultaneous estimation of the influx and efflux blood–brain barrier permeabilities of gabapentin using a microdialysis-pharmacokinetic approach. Pharm Res. 1996;13(3):398–403.
Carcaboso AM, Elmeliegy MA, Shen J, Juel SJ, Zhang ZM, Calabrese C, et al. Tyrosine kinase inhibitor gefitinib enhances topotecan penetration of gliomas. Cancer Res. 2010;70(11):4499–508.
Loos WJ, Zamboni WC, Engels FK, de Bruijn P, Lam MH, de Wit R, et al. Pitfalls of the application of microdialysis in clinical oncology: controversial findings with docetaxel. J Pharm Biomed Anal. 2007;45(2):288–94.
Whitaker G, Lunte CE. Investigation of microdialysis sampling calibration approaches for lipophilic analytes: doxorubicin. J Pharm Biomed Anal. 2010;53(3):490–6.
Lindberger M, Tomson T, Lars S. Microdialysis sampling of carbamazepine, phenytoin and phenobarbital in subcutaneous extracellular fluid and subdural cerebrospinal fluid in humans: an in vitro and in vivo study of adsorption to the sampling device. Pharmacol Toxicol. 2002;91(4):158–65.
Khramov AN, Stenken JA. Enhanced microdialysis recovery of some tricyclic antidepressants and structurally related drugs by cyclodextrin-mediated transport. Analyst. 1999;124(7):1027–33.
Elmeliegy MA, Carcaboso AM, Tagen M, Bai F, Stewart CF. Role of ATP-binding cassette and solute carrier transporters in Erlotinib CNS penetration and intracellular accumulation. Clin Cancer Res. 2011;17(1):89–99.
Müller M, Schmid R, Wagner O, Osten B, Shayganfar H, Eichler HG. In vivo characterization of transdermal drug transport by microdialysis. J Control Release. 1995;37(1–2):49–57.
Duo J, Fletcher H, Stenken JA. Natural and synthetic affinity agents as microdialysis sampling mass transport enhancers: current progress and future perspectives. Biosens Bioelectron. 2006;22(3):449–57.
Araujo BV, Silva CF, Haas SE, Dalla CT. Microdialysis as a tool to determine free kidney levels of voriconazole in rodents: a model to study the technique feasibility for a moderately lipophilic drug. J Pharm Biomed Anal. 2008;47(4–5):876–81.
Warner DL, Burke TG. Simple and versatile high-performance liquid chromatographic method for the simultaneous quantitation of the lactone and carboxylate forms of camptothecin anticancer drugs. J Chromatogr B Biomed Sci Appl. 1997;691(1):161–71.
Khramov AN, Stenken JA. Enhanced microdialysis extraction efficiency of ibuprofen in vitro by facilitated transport with beta-cyclodextrin. Anal Chem. 1999;71(7):1257–64.
Ao X, Stenken JA. Water-soluble cyclodextrin polymers for enhanced relative recovery of hydrophobic analytes during microdialysis sampling. Analyst. 2003;128(9):1143–9.
Menacherry S, Hubert W, Justice Jr JB. In vivo calibration of microdialysis probes for exogenous compounds. Anal Chem. 1992;64(6):577–83.
Jacobson I, Sandberg M, Hamberger A. Mass transfer in brain dialysis devices–a new method for the estimation of extracellular amino acids concentration. J Neurosci Methods. 1985;15(3):263–8.
D'argenio DZ, Schumitzky A. ADAPT 5 User’s guide: pharmacokinetic/ pharmacodynamic systems analysis software. Los Angeles: Biomedical Simulations Resource; 2009.
Schuck VJ, Rinas I, Derendorf H. In vitro microdialysis sampling of docetaxel. J Pharm Biomed Anal. 2004;36(4):807–13.
May M, Batkai S, Zoerner AA, Tsikas D, Jordan J, Engeli S. Enhanced human tissue microdialysis using hydroxypropyl-ss-cyclodextrin as molecular carrier. PLoS One. 2013;8(4):e60628.
Ward KW, Medina SJ, Portelli ST, Mahar Doan KM, Spengler MD, Ben MM, et al. Enhancement of in vitro and in vivo microdialysis recovery of SB-265123 using Intralipid and Encapsin as perfusates. Biopharm Drug Dispos. 2003;24(1):17–25.
Irie T, Uekama K. Pharmaceutical applications of cyclodextrins. III. Toxicological issues and safety evaluation. J Pharm Sci. 1997;86(2):147–62.
Cwiertnia B, Hladon T, Stobiecki M. Stability of diclofenac sodium in the inclusion complex with beta-cyclodextrin in the solid state. J Pharm pharmacol. 1999;51(11):1213–8.
Vangara KK, Ali HI, Lu D, Liu JL, Kolluru S, Palakurthi S. SN-38-cyclodextrin complexation and its influence on the solubility, stability, and in vitro anticancer activity against ovarian cancer. AAPS PharmSciTech. 2014;15(2):472–82.
Metz MZ, Gutova M, Lacey SF, Abramyants Y, Vo T, Gilchrist M, et al. Neural stem cell-mediated delivery of irinotecan-activating carboxylesterases to glioma: implications for clinical use. Stem Cells Transl Med. 2013;2(12):983–92.
Dodds HM, Tobin PJ, Stewart CF, Cheshire P, Hanna S, Houghton P, et al. The importance of tumor glucuronidase in the activation of irinotecan in a mouse xenograft model. J Pharmacol Exp Ther. 2002;303(2):649–55.
Ngim KK, Gu Z, Catalano T. Characterization and resolution of reversed phase HPLC chromatography failure attributed to sulfobutylether-beta-cyclodextrin in a pharmaceutical sample preparation. J Pharm Biomed Anal. 2009;49(3):660–9.
Hirayama F, Uekama K. Cyclodextrin-based controlled drug release system. Adv Drug Deliv Rev. 1999;36(1):125–41.
Zamboni WC, Houghton PJ, Hulstein JL, Kirstein M, Walsh J, Cheshire PJ, et al. Relationship between tumor extracellular fluid exposure to topotecan and tumor response in human neuroblastoma xenograft and cell lines. Cancer Chemother Pharmacol. 1999;43(4):269–76.
Dukic S, Kaltenbach ML, Gourdier B, Marty H, Vistelle R. Determination of free extracellular levels of methotrexate by microdialysis in muscle and solid tumor of the rabbit. Pharm Res. 1998;15(1):133–8.
Matsumura Y, Maeda H. A new concept for macromolecular therapeutics in cancer chemotherapy: mechanism of tumoritropic accumulation of proteins and the antitumor agent smancs. Cancer Res. 1986;46(12 Pt 1):6387–92.
Zhou Q, Lv H, Mazloom AR, Xu H, Ma'ayan A, Gallo JM. Activation of alternate prosurvival pathways accounts for acquired sunitinib resistance in U87MG glioma xenografts. J Pharmacol Exp Ther. 2012;343(2):509–19.
Evrard PA, Ragusi C, Boschi G, Verbeeck RK, Scherrmann JM. Simultaneous microdialysis in brain and blood of the mouse: extracellular and intracellular brain colchicine disposition. Brain Res. 1998;786(1–2):122–7.
Hammarlund-Udenaes M, Friden M, Syvanen S, Gupta A. On the rate and extent of drug delivery to the brain. Pharm Res. 2008;25(8):1737–50.
AMC acknowledges funding from the AECC Scientific Foundation, MINECO (SAF2011-22660), Fundacion BBVA, European Union Seventh Framework Programme (FP7/2007-2013) under Marie Curie International Reintegration Grant (PIRG-08-GA-2010-276998) and ISCIII-FEDER (CP13/00189). AS thanks the European Union's - Seventh Framework Programme under grant agreement #612675-MC-NANOTAR. Work supported by the Xarxa de Bancs de Tumors de Catalunya (XBTC) sponsored by Pla Director d’Oncologia de Catalunya. We thank Dr. Mireia Camos for performing erythrocyte counts.
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Monterrubio, C., Paco, S., Vila-Ubach, M. et al. Combined Microdialysis-Tumor Homogenate Method for the Study of the Steady State Compartmental Distribution of a Hydrophobic Anticancer Drug in Patient-Derived Xenografts. Pharm Res 32, 2889–2900 (2015). https://doi.org/10.1007/s11095-015-1671-9