title: Stability experiments in human urine with EO9 (apaziquone): a novel anticancer agent for the intravesical treatment of bladder cancer.
abstract: EO9 (apaziquone) is a novel, promising anticancer agent, which is currently being investigated for the intravesical treatment of bladder cancer. EO9 contains a highly reactive aziridine ring in its structure that limits its chemical stability in acidic aqueous solutions. The stability of the pharmaceutically formulated EO9 in human urine, including the effects of several parameters such as temperature, buffer strength and pH have been investigated. Urine extracts were analyzed by high-performance liquid chromatography coupled to electrospray tandem mass spectrometry (HPLC-MS/MS) using a TurboIonspray interface and positive-ion multiple reaction monitoring. EO9 was unstable in urine at 43 degrees C during the instillation for longer than 1 h. However, the drug was stable in human urine for 3 h at 37 degrees C. EO9 is stable in urine stabilized with TRIS buffer (pH 9.0; 5 mM) for up to three freeze/thaw cycles at -20 and -70 degrees C and 3 months of storage at -70 degrees C. The results also illustrated that with the lower pH in urine, EO9 became more unstable. Furthermore, a new degradation product of EO9 was discovered and successfully identified as EO9-Cl. The outcomes of these stability experiments will be implemented to insure proper sample handling at the clinical sites, transport, storage, and sample handling during analysis in the forthcoming preclinical studies of EO9 in superficial bladder cancer, supported by bioanalysis and pharmacokinetic monitoring.
authors.affiliation: Department of Pharmacy & Pharmacology, Slotervaart Hospital, The Netherlands Cancer Institute, Louwesweg 6, 1066 EC Amsterdam, The Netherlands. apliv@slz.nl
authors.forename: Liia D
authors.initials: LD
authors.lastname: Vainchtein
authors.forename: Hilde
authors.initials: H
authors.lastname: Rosing
authors.forename: Dorla
authors.initials: D
authors.lastname: Mirejovsky
authors.forename: Luigi
authors.initials: L
authors.lastname: Lenaz
authors.forename: Jan H M
authors.initials: JH
authors.lastname: Schellens
authors.forename: Jos H
authors.initials: JH
authors.lastname: Beijnen
mesh_terms: D000970:Antineoplastic Agents / Q000627:therapeutic use / Q000652:urine*; D001388:Aziridines / Q000627:therapeutic use / Q000652:urine*; D002021:Buffers; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D006863:Hydrogen-Ion Concentration; D045563:Indolequinones / Q000627:therapeutic use / Q000652:urine*; D013058:Mass Spectrometry; D012015:Reference Standards; D013048:Specimen Handling; D013696:Temperature; D001749:Urinary Bladder Neoplasms / Q000188:drug therapy*
chemical_list: D000970:Antineoplastic Agents; D001388:Aziridines; D002021:Buffers; D045563:Indolequinones; C060817:apaziquone