Investigating Intestinal Permeability of Bortezomib Using a Validated HPLC-UV Method

Abstract

Bortezomib (BTZ), as a proteasome inhibitor, has been used for treatment of patients with relapsed/refractory multiple myeloma and mantle cell lymphoma. BTZ is available for intravenous injection or subcutaneous administration. In this study, for evaluating the potential of BTZ oral delivery, intestinal permeability of BTZ was determined using in situ single-pass intestinal perfusion (SPIP) technique and the perfused solutions were analyzed using a validated HPLC-UV method. The chromatographic separation was performed using a C18 column via isocratic mode at a flow rate of 0.5 mL/min at 270 nm. The mobile phase was a mixture of methanol/deionized water (50:50% v/v) with 0.1% glacial acetic acid. The results indicated that calibration curves were linear (r² ˃0.99) in a concentration range of 1.65–5 µg/mL for BTZ and 8.33–25 µg/mL for phenol red. A limit of quantitation of 1.03 and 6.67 µg/mL was obtained for BTZ and phenol red, respectively. The recovery values were in the range of 96.5–105.4% for BTZ, and 88–99.2% for phenol red. The relative standard deviations (RSD) were ≤4.9% for BTZ and ≤7% for phenol red. Stability studies indicated that the working standard solution is stable over a period of 48 h at room temperature. Finally, an effective permeability (P_eff) value of (3.36±0.5)×10⁻⁵ cm/sec (mean±SEM) was achieved for BTZ. Moreover, it was predicted that BTZ belongs to the biopharmaceutical class ІІІ.

Key words

bortezomib - intestinal permeability - HPLC

• References

• 1 Adams J. The proteasome: A suitable antineoplastic target. Nature Reviews Cancer. 2004; 4: 349-360
• 2 Adams J, Palombella VJ, Sausville EA. et al. Proteasome inhibitors: A novel class of potent and effective antitumor agents. Cancer Research. 1999; 59: 2615-2622
• 3 Orlowski RZ, Kuhn DJ. Proteasome inhibitors in cancer therapy: Lessons from the first decade. Clinical Cancer Research. 2008; 14: 1649-1657
• 4 LeBlanc R, Catley LP, Hideshima T. et al. Proteasome inhibitor PS-341 inhibits human myeloma cell growth in vivo and prolongs survival in a murine model. Cancer Research. 2002; 62: 4996-5000
• 5 Sunwoo JB, Chen Z, Dong G. et al. Novel proteasome inhibitor PS-341 inhibits activation of nuclear factor-κB, cell survival, tumor growth, and angiogenesis in squamous cell carcinoma. Clinical Cancer Research. 2001; 7: 1419-1428
• 6 Shah SA, Potter MW, McDade TP. et al. 26S proteasome inhibition induces apoptosis and limits growth of human pancreatic cancer. Journal of Cellular Biochemistry. 2001; 82: 110-122
• 7 Frankel A, Man S, Elliott P. et al. Lack of multicellular drug resistance observed in human ovarian and prostate carcinoma treated with the proteasome inhibitor PS-341. Clinical Cancer Research. 2000; 6: 3719-3728
• 8 Imajohohmi S, Kawaguchi T, Sugiyama S. et al. Lactacystin, a specific inhibitor of the proteasome, induces apoptosis in human monoblast U937 cells. Biochemical and Biophysical Research Communications 1995; 217: 1070-1077
• 9 Chen D, Frezza M, Schmitt S. et al. Bortezomib as the first proteasome inhibitor anticancer drug: Current status and future perspectives. Current Cancer Drug Targets. 2011; 11: 239
• 10 Bold R. “Development of the Proteasome Inhibitor Velcade™(Bortezomib)” by Julian Adams, Ph. D., and Michael Kauffman, MD, Ph. D. Cancer Investigation 2004; 22: 328-329
• 11 Pennisi A, Li X, Ling W. et al. The proteasome inhibitor, bortezomib suppresses primary myeloma and stimulates bone formation in myelomatous and nonmyelomatous bones in vivo. American Journal of Hematology. 2009; 84: 6-14
• 12 Mazzaferro S, Bouchemal K, Ponchel G. Oral delivery of anticancer drugs I: general considerations. Drug Discovery Today. 2013; 18: 25-34
• 13 Mei L, Zhang Z, Zhao L. et al. Pharmaceutical nanotechnology for oral delivery of anticancer drugs. Advanced Drug Delivery Reviews. 2013; 65: 880-890
• 14 Thanki K, Gangwal RP, Sangamwar AT. et al. Oral delivery of anticancer drugs: challenges and opportunities. Journal of Controlled Release. 2013; 170: 15-40
• 15 Bardelmeijer HA, van Tellingen O, Schellens JH. et al. The oral route for the administration of cytotoxic drugs: Strategies to increase the efficiency and consistency of drug delivery. Investigational New Drugs. 2000; 18: 231-241
• 16 Varma MV, Khandavilli S, Ashokraj Y. et al. Biopharmaceutic classification system: A scientific framework for pharmacokinetic optimization in drug research. Current Drug Metabolism. 2004; 5: 375-388
• 17 Veber DF, Johnson SR, Cheng H-Y. et al. Molecular properties that influence the oral bioavailability of drug candidates. Journal of Medicinal Chemistry 2002; 45: 2615-2623
• 18 Aungst BJ. Intestinal permeation enhancers. Journal of pharmaceutical sciences 2000; 89: 429-442
• 19 Incecayir T, Tsume Y, Amidon GL. Comparison of the permeability of metoprolol and labetalol in rat, mouse, and Caco-2 cells: Use as a reference standard for BCS classification. Molecular Pharmaceutics. 2013; 10: 958-966
• 20 Zur M, Gasparini M, Wolk O. et al. The low/high BCS permeability class boundary: Physicochemical comparison of metoprolol and labetalol. Molecular Pharmaceutics. 2014; 11: 1707-1714
• 21 Gupta D, Varghese Gupta S, Dahan A. et al. Increasing oral absorption of polar neuraminidase inhibitors: A prodrug transporter approach applied to oseltamivir analogue. Molecular Pharmaceutics. 2013; 10: 512-522
• 22 Cao F, Jia J, Yin Z. et al. Ethylene glycol-linked amino acid diester prodrugs of oleanolic acid for PepT1-mediated transport: Synthesis, intestinal permeability and pharmacokinetics. Molecular Pharmaceutics. 2012; 9: 2127-2135
• 23 Huber L. Validation of analytical methods. Agilent Technologies 2010; 13-28
• 24 Olfert ED, Cross BM, McWilliam AA. Guide to the care and use of experimental animals: Canadian council on animal care ottawa. vol 1, 1993
• 25 Islambulchilar Z, Ghanbarzadeh S, Emami S. et al. Development and validation of an HPLC method for the analysis of sirolimus in drug products. Advanced Pharmaceutical Bulletin 2012; 2: 135-139
• 26 Zakeri-Milani P, Valizadeh H, Azarmi Y. et al. Simultaneous determination of metoprolol, propranolol and phenol red in samples from rat in situ intestinal perfusion studies. DARU Journal of Pharmaceutical Sciences 2006; 14: 102-108
• 27 Salphati L, Childers K, Pan L. et al. Evaluation of a single-pass intestinal-perfusion method in rat for the prediction of absorption in man. Journal of Pharmacy and Pharmacology 2001; 53: 1007-1013
• 28 Song N-N, Li Q-S, Liu C-X. Intestinal permeability of metformin using single-pass intestinal perfusion in rats. World Journal of Gastroenterology 2006; 12: 4064
• 29 Zakeri-Milani P, Valizadeh H, Islambulchilar Z. et al. Investigation of the intestinal permeability of ciclosporin using the in situ technique in rats and the relevance of P-glycoprotein. Arzneimittel-Forschung 2008; 58: 188-192
• 30 Zakeri-Milani P, Valizadeh H, Tajerzadeh H. et al. The utility of rat jejunal permeability for biopharmaceutics classification system. Drug Development and Industrial Pharmacy. 2009; 35: 1496-1502
• 31 Suh KS, Goy A. Bortezomib in mantle cell lymphoma. Future Oncology 2008; 4: 149-168
• 32 Zakeri-Milani P, Valizadeh H, Tajerzadeh H. et al. Predicting human intestinal permeability using single-pass intestinal perfusion in rat. Journal of Pharmacy and Pharmaceutical Siences 2007; 10: 368-379