S F Stinson1, K Hill, T J Siford, L R Phillips, T W Daw. 1. Laboratory of Drug Discovery Research and Development, Developmental Therapeutics Program, Division of Cancer Treatment Diagnosis and Centers, National Cancer Institute-FCRDC, Frederick, Maryland 21702, USA.
Abstract
PURPOSE: Flavopiridol is a flavone which inhibits several cyclin-dependent kinases, and exhibits potent growth-inhibitory activity against a number of human tumor cell lines both in vitro, and when grown as xenografts in mice. It is currently being evaluated in a phase I clinical trial at the National Cancer Institute. The objective of this project was to develop and validate an analytical method for the assay of flavopiridol in human plasma, with sufficient sensitivity to permit the plasma pharmacokinetics of flavopiridol to be studied during clinical trials. METHODS: Flavopiridol was isolated from human plasma samples by extraction with t-butylmethyl ether following alkalinization with borate buffer (pH 8.0). The extract was evaporated, the residue was dissolved in mobile phase, and analyzed by reversed-phase high-pressure liquid chromatography. Chromatography was accomplished with a polymer-based C18 column eluted with a mobile phase consisting of methanol-phosphate buffer, pH 11.0 (53:47 v/v). Electrochemical detection (ECD) was employed. RESULTS: Flavopiridol was recovered from human plasma with an efficiency of 85-87%. Calibration curves were linear over the concentration range 10-500 nM (4.4-219 ng/ml). Plasma standard concentrations were measured with an accuracy and precision ranging from 3.2% to 10%. Regression analysis of flavopiridol concentrations of 15 clinical trial plasma samples ranging in concentration from approximately 50 to 4000 microM quantitated by both ECD and mass spectrometry showed close agreement. The equation of the regression line was y = 1.02x + 8 with a correlation coefficient of 0.969. Continuous infusion of flavopiridol in four patients for 72 h at a rate of 50 mg/m2 per day, resulted in mean steady-state plasma concentrations of from 200 to 300 nM. Levels declined in a biexponential manner following termination of the infusion, falling to approximately 10 nM after 48 h. CONCLUSIONS: An analytical method for the assay of flavopiridol in human plasma was developed with sensitivity to at least 10 nM. The assay is accurate, precise and specific, and is suitable for determination of plasma flavopiridol concentrations for pharmacokinetic studies during clinical trials.
PURPOSE:Flavopiridol is a flavone which inhibits several cyclin-dependent kinases, and exhibits potent growth-inhibitory activity against a number of humantumor cell lines both in vitro, and when grown as xenografts in mice. It is currently being evaluated in a phase I clinical trial at the National Cancer Institute. The objective of this project was to develop and validate an analytical method for the assay of flavopiridol in human plasma, with sufficient sensitivity to permit the plasma pharmacokinetics of flavopiridol to be studied during clinical trials. METHODS:Flavopiridol was isolated from human plasma samples by extraction with t-butylmethyl ether following alkalinization with borate buffer (pH 8.0). The extract was evaporated, the residue was dissolved in mobile phase, and analyzed by reversed-phase high-pressure liquid chromatography. Chromatography was accomplished with a polymer-based C18 column eluted with a mobile phase consisting of methanol-phosphate buffer, pH 11.0 (53:47 v/v). Electrochemical detection (ECD) was employed. RESULTS:Flavopiridol was recovered from human plasma with an efficiency of 85-87%. Calibration curves were linear over the concentration range 10-500 nM (4.4-219 ng/ml). Plasma standard concentrations were measured with an accuracy and precision ranging from 3.2% to 10%. Regression analysis of flavopiridol concentrations of 15 clinical trial plasma samples ranging in concentration from approximately 50 to 4000 microM quantitated by both ECD and mass spectrometry showed close agreement. The equation of the regression line was y = 1.02x + 8 with a correlation coefficient of 0.969. Continuous infusion of flavopiridol in four patients for 72 h at a rate of 50 mg/m2 per day, resulted in mean steady-state plasma concentrations of from 200 to 300 nM. Levels declined in a biexponential manner following termination of the infusion, falling to approximately 10 nM after 48 h. CONCLUSIONS: An analytical method for the assay of flavopiridol in human plasma was developed with sensitivity to at least 10 nM. The assay is accurate, precise and specific, and is suitable for determination of plasma flavopiridol concentrations for pharmacokinetic studies during clinical trials.
Authors: John C Byrd; Thomas S Lin; James T Dalton; Di Wu; Mitch A Phelps; Beth Fischer; Mollie Moran; Kristie A Blum; Brad Rovin; Michelle Brooker-McEldowney; Sarah Broering; Larry J Schaaf; Amy J Johnson; David M Lucas; Nyla A Heerema; Gerard Lozanski; Donn C Young; Jose-Ramon Suarez; A Dimitrios Colevas; Michael R Grever Journal: Blood Date: 2006-09-26 Impact factor: 22.113
Authors: Qing Liu; Katherine L Farley; Amy J Johnson; Natarajan Muthusamy; Craig C Hofmeister; Kristie A Blum; Larry J Schaaf; Michael R Grever; John C Byrd; James T Dalton; Mitch A Phelps Journal: Ther Drug Monit Date: 2008-10 Impact factor: 3.681
Authors: Luis M Schang; Andrew Bantly; Marie Knockaert; Farida Shaheen; Laurent Meijer; Michael H Malim; Nathanael S Gray; Priscilla A Schaffer Journal: J Virol Date: 2002-08 Impact factor: 5.103
Authors: Mitch A Phelps; Darlene M Rozewski; Jeffrey S Johnston; Katherine L Farley; Katie A Albanese; John C Byrd; Thomas S Lin; Michael R Grever; James T Dalton Journal: J Chromatogr B Analyt Technol Biomed Life Sci Date: 2008-04-22 Impact factor: 3.205
Authors: J H Hooijberg; H J Broxterman; G L Scheffer; C Vrasdonk; M Heijn; M C de Jong; R J Scheper; J Lankelma; H M Pinedo Journal: Br J Cancer Date: 1999-09 Impact factor: 7.640