title: Overcoming bioanalytical challenges associated with the separation and quantitation of GSK1278863, a HIF-prolyl hydroxylase inhibitor, and its 14 stereoisomeric metabolites.
abstract: GSK1278863 is an investigative drug under investigation for treatment of anemia associated with chronic kidney disease. Its metabolism is primarily metabolized by P450 enzymes where 19 unique metabolic species have been identified. These include multiple products of mono-, di-, and tri-oxygenation. Initially, two separate and complex ultra high performance liquid chromatography (UHPLC) reverse phase methodologies were developed, validated and applied to measure parent and various predominant and circulating metabolites in numerous clinical studies. However, 5 of the 6 oxidative metabolites may exist in different stereoisomeric forms, resulting in 14 separate species; therefore a chiral methodology was required to determine which stereoisomeric forms circulated in human. A variety of conventional approaches were explored, where in the end a supercritical fluid chromatography (SFC) method was required to separate this complex mixture of 14 stereoisomeric metabolites; data from these experiments provided important information on which species circulate in human. The details of these methodologies will be discussed herein.
authors.affiliation: Bioanalytical Science and Toxicokinetics, Platform Science and Technology, GlaxoSmithkline Pharmaceuticals, 709 Swedeland Road, King of Prussia, PA 19406, USA. Electronic address: hermes.2.licea-perez@gsk.com.
authors.forename: Hermes
authors.initials: H
authors.lastname: Licea Perez
authors.affiliation: Bioanalytical Science and Toxicokinetics, Platform Science and Technology, GlaxoSmithkline Pharmaceuticals, 709 Swedeland Road, King of Prussia, PA 19406, USA.
authors.forename: Dana
authors.initials: D
authors.lastname: Knecht
authors.affiliation: Bioanalytical Science and Toxicokinetics, Platform Science and Technology, GlaxoSmithkline Pharmaceuticals, 709 Swedeland Road, King of Prussia, PA 19406, USA.
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Evans
mesh_terms: D001463:Barbiturates / Q000097:blood* / Q000302:isolation & purification / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D056148:Chromatography, Reverse-Phase / Q000379:methods; D025924:Chromatography, Supercritical Fluid / Q000379:methods*; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D005998:Glycine / Q000031:analogs & derivatives* / Q000097:blood / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D010084:Oxidation-Reduction; D064800:Prolyl-Hydroxylase Inhibitors / Q000097:blood* / Q000302:isolation & purification / Q000378:metabolism; D015203:Reproducibility of Results; D013237:Stereoisomerism
chemical_list: D001463:Barbiturates; C000599718:GSK1278863; D064800:Prolyl-Hydroxylase Inhibitors; D003577:Cytochrome P-450 Enzyme System; D005998:Glycine
keywords: GSK1278863; High performance fraction collector LCJet system; Stereoisomeric metabolite separation; Supercritical fluid chromatography (SFC); Ultra high performance liquid chromatography tandem mass spectrometry (UHPLC–MS/MS); Ultra performance convergence chromatography (UPC(2))