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. 2007 Aug 3;8(3):E78–E87. doi: 10.1208/pt0803061

Development of a bladder instillation of the indoloquinone anticancer agent EO-9 using tert-butyl alcohol as lyophilization vehicle

Sabien C van der Schoot 1,✉, Bastiaan Nuijen 1,✉, Frits M Flesch 2, Ashok Gore 3, Dorla Mirejovsky 3, Luigi Lenaz 3, Jos H Beijnen 1,2
PMCID: PMC2750557  PMID: 17915811

Abstract

The purpose of this research was to develop a stable bladder instillation of EO-9 for the treatment of superficial bladder cancer. First, stability and dissolution studies were performed. Subsequently, the freeze-drying process was optimized by determination of the freeze-drying characteristics of the selected cosolvent/water system and differential scanning calorimetry analysis of the formulation solution. Furthermore, the influence of the freeze-drying process on crystallinity and morphology of the freeze-dried product was determined with x-ray diffraction analysis and scanning electron microscopy, respectively. Subsequently, a reconstitution solution was developed. This study revealed that tert-butyl alcohol (TBA) can be used to both dramatically improve the solubility and stability of EO-9 and to shorten the freeze-drying cycle by increasing the sublimation rate. During freeze drying, 3 TBA crystals were found: TBA hydrate-ice crystals, crystals of TBA hydrate, and a third crystal, probably composed of TBA hydrate crystals containing ≈90% to 95% TBA. Furthermore, it was shown that crystallization of TBA hydrate was inhibited in the presence of both sodium bicarbonate (NaHCO3) and mannitol. Addition of an annealing step resulted in a minor increase in the crystallinity of the freeze-dried product and formation of the δ-polymorph of mannitol. A stable bladder instillation was obtained after reconstitution of the freeze-dried product (containing 8 mg of EO-9, 20 mg of NaHCO3, and 50 mg of mannitol per vial) to 20 mL with a reconstitution solution composed of propylene glycol/water for injection (WfI)/NaHCO3/sodium edetate 60%/40%/2%/0.02% vol/vol/wt/wt, followed by dilution with Wfl to a final volume of 40 mL.

Keywords: Mitomycin analogue, EO-9, formulation development, freeze drying, tert-butyl alcohol

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References

  • 1.Bailey SM, Lewis AD, Patterson LH, Fisher GR, Knox RJ, Workman P. Involvement of NADPH: cytochrome P450 reductase in the activation of indoloquinone EO9 to free radical and DNA damaging species. Biochem Pharmacol. 2001;62:461–468. doi: 10.1016/S0006-2952(01)00674-8. [DOI] [PubMed] [Google Scholar]
  • 2.Walton MI, Smith PJ, Workman P. The role of NAD(P)H: quinone reductase (EC 1.6.99.2, DT-diaphorase) in the reductive bioactivation of the novel indoloquinone antitumor agent EO9. Cancer Commun. 1991;3:199–206. doi: 10.3727/095535491820873164. [DOI] [PubMed] [Google Scholar]
  • 3.Hendriks HR, Pizao PE, Berger DP, et al. EO9: a novel bioreductive alkylating indo loquinone with preferential solid tumour activity and lack of bone marrow toxicity in preclinical models. Eur J Cancer. 1993;29:897–906. doi: 10.1016/S0959-8049(05)80434-4. [DOI] [PubMed] [Google Scholar]
  • 4.Dass CR. Improving anti-angiogenic therapy via selective delivery of cationic liposomes to tumour vasculature. Int J Pharm. 2003;267:1–12. doi: 10.1016/j.ijpharm.2003.08.010. [DOI] [PubMed] [Google Scholar]
  • 5.Jonkman-de Vries JD, Talsma H, Henrar REC, Kettenes.-van den Bosch JJ, Bult A, Beijnen JH. Pharmaceutical formulation development of a parenteral lyophilized formulation of the novel indoloquinone antitumor, agent EO9. Cancer Chemother Pharmacol. 1994;34:416–422. doi: 10.1007/BF00685567. [DOI] [PubMed] [Google Scholar]
  • 6.Dirix LY, Tonnesen F, Cassidy J, et al. EO9 phase II study in advanced breast, gastric, pancreatic and colorectal carcinoma by the EORTC, Early Clinical Studies Group. Eur J Cancer. 2007;32:2019–2022. doi: 10.1016/0959-8049(96)00226-2. [DOI] [PubMed] [Google Scholar]
  • 7.Schellens JH, Dombernowsky P, Cassidy J, et al. Population pharmacokinetics and dynamics in phase II studies of the novel bioreductive alkylating cytotoxic indoloquinone EO9. Anticancer Drugs. 2001;12:583–590. doi: 10.1097/00001813-200108000-00004. [DOI] [PubMed] [Google Scholar]
  • 8.Phillips RM, Hulbert PB, Bibby MC, Sleigh NR, Double JA. In vitro activity of the novel indoloquinone EO-9 and the influence of pH on cytotoxicity. Br J Cancer. 1992;65:359–364. doi: 10.1038/bjc.1992.73. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9.Teagarden DL, Baker DS. Practical aspects of lyophilization using non-aqueous co-solvent systems. Eur J Pharm Sci. 2002;15:115–133. doi: 10.1016/S0928-0987(01)00221-4. [DOI] [PubMed] [Google Scholar]
  • 10.Ni N, Tesconi M, Tabibi SE, Gupta S, Yalkowsky SH. Use of pure t-butanol as a solvent for freeze-drying: a case study. Int. J Pharm. 2001;226:39–46. doi: 10.1016/S0378-5173(01)00757-8. [DOI] [PubMed] [Google Scholar]
  • 11.Kasraian K, DeLuca PP. Thermal analysis of the tertiary butyl alcohol-water system and its implications on freeze-drying. Pharm Res. 1995;12:484–490. doi: 10.1023/A:1016233408831. [DOI] [PubMed] [Google Scholar]
  • 12.Wittaya-Areekul S, Nail SL. Freeze-drying of tert-butyl alcohol/water cosolvent systems: effects of formulation and process variables on residual solvents. J Pharm Sci. 1998;87:491–495. doi: 10.1021/js9702832. [DOI] [PubMed] [Google Scholar]
  • 13.Wittaya-Areekul S, Needham GF, Milton N, Roy ML, Nail SL. Freeze-drying of tert-butanol/water cosolvent systems: a case report on formation of a friable freeze-dried powder of tobramycin sulfate. J Pharm Sci. 2002;91:1147–1155. doi: 10.1002/jps.10113. [DOI] [PubMed] [Google Scholar]
  • 14.Kasraian K, DeLuca PP. The effect of tertiary butyl alcohol on the resistance of the dry product layer during primary drying. Pharm Res. 1995;12:491–495. doi: 10.1023/A:1016285425670. [DOI] [PubMed] [Google Scholar]
  • 15.Nuijen B, Bouma M, Henrar REC, et al. Pharmaceutical development of a parenteral lyophilized formulation of the novel antitumor agent aplidine. PDA J Pharm Sci Technol. 2000;54:193–208. [PubMed] [Google Scholar]
  • 16.de Vries JD, Winkelhorst J, Underberg WJM, Henrar REC, Beijnen JH. A systematic study on the chemical stability of the novel indoloquinone antitumor agent EO9. Int J Pharm. 1993;100:181–188. doi: 10.1016/0378-5173(93)90089-X. [DOI] [Google Scholar]
  • 17.Au JLS, Badalament RA, Guillaume Wientjes M, et al. Methods to improve efficacy of intravesical mitomycin C: results of a randomized phase III trial. J Natl Cancer Inst. 2001;93:597–604. doi: 10.1093/jnci/93.8.597. [DOI] [PubMed] [Google Scholar]
  • 18.Telang C, Suryanarayanan R. Crystallization of cephalothin sodium during lyophilization from tert-butyl alcohol-water cosolvent system. Pharm Res. 2005;22:153–160. doi: 10.1007/s11095-004-9021-3. [DOI] [PubMed] [Google Scholar]
  • 19.Telang C, Yu L, Suryanarayanan R. Effective inhibition of mannitol crystallization in frozen solutions by sodium chloride. Pharm Res. 2003;20:660–667. doi: 10.1023/A:1023263203188. [DOI] [PubMed] [Google Scholar]
  • 20.Yu L, Milton N, Groleau EG, Mishra DS, Vansickle RE. Existence of a mannitol hydrate during freeze-drying and practical implications. J Pharm Sci. 1999;88:196–198. doi: 10.1021/js980323h. [DOI] [PubMed] [Google Scholar]
  • 21.Lindamood C, Farnell DR, Giles HD, et al. Subchronic toxicity studies of t-butyl alcohol in rats and mice. Fundam Appl Toxicol. 1992;19:91–100. doi: 10.1016/0272-0590(92)90032-D. [DOI] [PubMed] [Google Scholar]
  • 22.Pikal MJ, Shah S, Roy ML, Putman R. The secondary drying stage of freeze drying: drying kinetics as a function of temperature and chamber pressure. Int J Pharm. 1990;60:203–217. doi: 10.1016/0378-5173(90)90074-E. [DOI] [Google Scholar]
  • 23.Impurities: Guideline for Residual Solvents. ICH Topic Q3C. International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use; Gaithersburg, MD: NIST; 1997
  • 24.Telang C, Suryanarayanan R, Yu L. Crystallization of D-mannitol in binary mixtures with NaCl: phase diagram and polymorphism. Pharm Res. 2003;20:1939–1945. doi: 10.1023/B:PHAM.0000008040.14958.bc. [DOI] [PubMed] [Google Scholar]

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