title: High-performance liquid chromatographic method for sensitive determination of the alkylating agent CB1954 in human plasma.
abstract: A high-performance liquid chromatography (HPLC) method is described for the measurement of the weak alkylating agent CB1954 in human plasma. CB1954 can be used as an innocuous prodrug designed for activation by bacterial nitroreductases in strategies of gene-directed enzyme-prodrug therapy, and becomes activated to a potent bifunctional alkylating agent. The HPLC method involves precipitation and solvent extraction and uses Mitomycin C (MMC) as an internal standard, with a retention time for MMC of 5.85 +/- 0.015 min, and for CB1954 of 10.72 +/- 0.063 min. The limit of detection for CB1954 is 2.9 ng/ml, and this compares favourably with systems involving direct analysis of plasma (limit of detection 600 ng/ml, approximately). The method is now being used for pharmacokinetic measurements in plasma samples from cancer patients entering phase I clinical trials of CB1954. Results using serial plasma samples from one patient are presented. The patient was treated intravenously with CB1954 (6 mg/m2), and plasma clearance of the drug showed biphasic kinetics with alpha half-life 14.6 min, and beta half-life 170.5 min.
authors.affiliation: CRC Institute for Cancer Studies, University of Birmingham, UK.
authors.forename: D
authors.initials: D
authors.lastname: Anderson
authors.forename: D R
authors.initials: DR
authors.lastname: Ferry
authors.forename: R J
authors.initials: RJ
authors.lastname: Knox
authors.forename: S J
authors.initials: SJ
authors.lastname: Andrews
authors.forename: A J
authors.initials: AJ
authors.lastname: Downes
authors.forename: D J
authors.initials: DJ
authors.lastname: Kerr
authors.forename: L W
authors.initials: LW
authors.lastname: Seymour
mesh_terms: D018906:Antineoplastic Agents, Alkylating / Q000097:blood* / Q000493:pharmacokinetics; D001388:Aziridines / Q000097:blood* / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet
chemical_list: D018906:Antineoplastic Agents, Alkylating; D001388:Aziridines; C100099:tretazicar