title: Development of a bladder instillation of the indoloquinone anticancer agent EO-9 using tert-butyl alcohol as lyophilization vehicle.
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 approximately 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 delta-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 WfI to a final volume of 40 mL.
authors.affiliation: Department of Pharmacy and Pharmacology, Slotervaart Hospital/The Netherlands Cancer Institute, Louwesweg 6, 1066 EC Amsterdam, The Netherlands.
authors.forename: Sabien C
authors.initials: SC
authors.lastname: van der Schoot
authors.forename: Bastiaan
authors.initials: B
authors.lastname: Nuijen
authors.forename: Frits M
authors.initials: FM
authors.lastname: Flesch
authors.forename: Ashok
authors.initials: A
authors.lastname: Gore
authors.forename: Dorla
authors.initials: D
authors.lastname: Mirejovsky
authors.forename: Luigi
authors.initials: L
authors.lastname: Lenaz
authors.forename: Jos H
authors.initials: JH
authors.lastname: Beijnen
mesh_terms: D000283:Administration, Intravesical; D000970:Antineoplastic Agents / Q000008:administration & dosage*; D001388:Aziridines / Q000008:administration & dosage* / Q000737:chemistry; D002152:Calorimetry, Differential Scanning; D002849:Chromatography, Gas; D002851:Chromatography, High Pressure Liquid; D004355:Drug Stability; D005612:Freeze Drying; D045563:Indolequinones / Q000008:administration & dosage* / Q000737:chemistry; D008855:Microscopy, Electron, Scanning; D014677:Pharmaceutical Vehicles; D012995:Solubility; D014961:X-Ray Diffraction; D020002:tert-Butyl Alcohol / Q000008:administration & dosage*
chemical_list: D000970:Antineoplastic Agents; D001388:Aziridines; D045563:Indolequinones; D014677:Pharmaceutical Vehicles; C060817:apaziquone; D020002:tert-Butyl Alcohol