title: Determination of CH330331, a novel 4-anilinoquinazoline inhibitor of epidermal growth factor receptor tyrosine kinase, in human Caco-2 monolayers by high performance liquid chromatography with ultraviolet detection: application to a trans-epithelial transport study.
abstract: 4-Anilinoquinazolines (e.g. Iressa and Glivec) are a class of epidermal growth factor receptor tyrosine kinase (EGFR-TK) inhibitors widely used to treat non-small cell lung cancer and other tumors. However, low clinical response rate, resistance, and host toxicity of currently available EGFR-TK inhibitors prompt the development of second generation of TK inhibitors with improved efficacy, selectivity, and less resistance. CH330331 is a recently synthesized novel 4-anilinoquinazoline analog with confirmed anticancer activity in vitro and in vivo. To predict its oral pharmacokinetic behavior and transport nature in the intestine before entering clinical trials, we have developed and validated a high performance liquid chromatographic (HPLC) method for the determination of CH330331 in Caco-2 (a human colon cancer cell line) monolayers. The developed HPLC method was sensitive and reliable, with acceptable accuracy (90-110% of nominal values) and precision (intra- and inter-assay R.S.D.<10%). The total running time was within 10 min, with acceptable separation of the target analytes. The lower limit of quantitation (LLOQ) value for CH330331 was 200 ng/ml when an aliquot of 100 microl sample was injected onto the HPLC. The validated HPLC method was applied to characterize the epithelial transport of CH330331 in Caco-2 monolayers. The transport of CH330331 across the Caco-2 monolayers from the apical to basolateral side was 8- to 10-fold higher than that from the basolateral to apical side. Co-incubation of sodium azide or MK-571, but not verapamil, significantly inhibited the apical to basolateral transport of CH330331. These findings provide initial evidence that the intestinal absorption of CH330331 is mediated by an active mechanism. Further studies are required to explore the interaction of CH330331 with ATP-binding cassette transporters and the possible influence on its pharmacokinetics and pharmacodynamics.
authors.affiliation: Institute of Clinical Pharmacology, School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou 510080, China.
authors.forename: Hai-Yan
authors.initials: HY
authors.lastname: Sun
authors.forename: Su
authors.initials: S
authors.lastname: Guan
authors.forename: Hui-Chang
authors.initials: HC
authors.lastname: Bi
authors.forename: Qi-Biao
authors.initials: QB
authors.lastname: Su
authors.forename: Wen-Lin
authors.initials: WL
authors.lastname: Huang
authors.forename: Balram
authors.initials: B
authors.lastname: Chowbay
authors.forename: Min
authors.initials: M
authors.lastname: Huang
authors.forename: Xiao
authors.initials: X
authors.lastname: Chen
authors.forename: Chun-Guang
authors.initials: CG
authors.lastname: Li
authors.forename: Shu-Feng
authors.initials: SF
authors.lastname: Zhou
mesh_terms: D001692:Biological Transport; D018938:Caco-2 Cells; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004848:Epithelium / Q000378:metabolism; D066246:ErbB Receptors / Q000037:antagonists & inhibitors*; D006801:Humans; D047428:Protein Kinase Inhibitors / Q000493:pharmacokinetics / Q000494:pharmacology*; D011799:Quinazolines / Q000493:pharmacokinetics / Q000494:pharmacology*; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*
chemical_list: C523326:CH330331 compound; D047428:Protein Kinase Inhibitors; D011799:Quinazolines; D066246:ErbB Receptors