Development of a robust, sensitive and selective liquid chromatography-tandem mass spectrometry assay for the quantification of the novel macrocyclic peptide kappa opioid receptor antagonist [D-Trp]CJ-15,208 in plasma and application to an initial pharmacokinetic study.
Selective kappa opioid receptor (KOR) antagonists may have therapeutic potential as treatments for substance abuse and mood disorders. Since [D-Trp]CJ-15,208 (cyclo[Phe-d-Pro-Phe-d-Trp]) is a novel potent KOR antagonist in vivo, it is imperative to evaluate its pharmacokinetic properties to assist the development of analogs as potential therapeutic agents, necessitating the development and validation of a quantitative method for determining its plasma levels. A method for quantifying [D-Trp]CJ-15,208 was developed employing high performance liquid chromatography-tandem mass spectrometry in mouse plasma. Sample preparation was accomplished through a simple one-step protein precipitation method with acetonitrile, and [D-Trp]CJ-15,208 analyzed following HPLC separation on a Hypersil BDS C8 column. Multiple reaction monitoring (MRM), based on the transitions m/z 578.1→217.1 and 245.0, was specific for [D-Trp]CJ-15,208, and MRM based on the transition m/z 566.2→232.9 was specific for the internal standard without interference from endogenous substances in blank mouse plasma. The assay was linear over the concentration range 0.5-500ng/mL with a mean r(2)=0.9987. The mean inter-day accuracy and precision for all calibration standards were 93-118% and 8.9%, respectively. The absolute recoveries were 85±6% and 81±9% for [D-Trp]CJ-15,208 and the internal standard, respectively. The analytical method had excellent sensitivity with a lower limit of quantification of 0.5ng/mL using a sample volume of 20μL. The method was successfully applied to an initial pharmacokinetic study of [D-Trp]CJ-15,208 following intravenous administration to mice.
Department of Medicinal Chemistry, the University of Kansas, Lawrence, KS 66045, USA; Department of Medicinal Chemistry, University of Florida, Gainesville, FL 32610, USA.
Tanvir
T
Khaliq
Mass Spectrometry and Analytical Proteomics Laboratory, the University of Kansas, Lawrence, KS 66045, USA.
Todd D
TD
Williams
Department of Medicinal Chemistry, the University of Kansas, Lawrence, KS 66045, USA.
Sanjeewa N
SN
Senadheera
Department of Medicinal Chemistry, the University of Kansas, Lawrence, KS 66045, USA; Department of Medicinal Chemistry, University of Florida, Gainesville, FL 32610, USA. Electronic address: janealdrich@ufl.edu.
Jane V
JV
Aldrich
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D057230:Limit of Detection; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009292:Narcotic Antagonists / Q000097:blood*; D010456:Peptides, Cyclic / Q000097:blood*; D017473:Receptors, Opioid, kappa / Q000037:antagonists & inhibitors*; D053719:Tandem Mass Spectrometry / Q000379:methods*
C540936:CJ 15,208; D009292:Narcotic Antagonists; D010456:Peptides, Cyclic; D017473:Receptors, Opioid, kappa
Kappa opioid receptor antagonist; LC–MS/MS; Macrocyclic peptide; Multiple reaction monitoring; Peptide quantitation; [D-Trp]CJ-15,208