Synthesis, characterisation, and mass spectrometric detection of a pegylated EPO-mimetic peptide for sports drug testing purposes

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Bibliographic Details
Title translated into German:Synthese, Charakterisierung und massenspektrometrischer Nachweis eines pegylierten EPO-mimetischen Peptids zu Doping-Testzwecken
Author:Möller, Ines; Thomas, Andreas; Geyer, Hans; Schänzer, Wilhelm; Thevis, Mario
Published in:Rapid communications in mass spectrometry
Published:25 (2011), 15, S. 2115-2123, Lit.
Format: Publications (Database SPOLIT)
Publication Type: Journal article
Media type: Electronic resource (online) Print resource
Language:English
ISSN:0951-4198
Keywords:
Online Access:
Identification number:PU201410009726
Source:BISp

Author's abstract

Erythropoietin (EPO) and other erythropoiesis-stimulating agents possess a high misuse potential in elite sport due to their ability to increase the oxygen transport capacity, which plays a vital role in enhancing endurance performance. Currently, a new generation of EPO-mimetic peptides is under development from which peginesatide (also referred to as Hematide™), a pegylated homodimeric peptide of approximately 45 kDa with no structural relationship to erythropoietin, is the most advanced drug candidate undergoing phase-III clinical trials. Since preventive doping research aims at the development of detection methods before a drug receives clinical approval, a selective and sensitive assay has to be established knowing that conventional doping control assays for EPO will not succeed in detecting peginesatide. Thus, a pegylated EPO-mimetic peptide simulating the structure and properties of the lead drug candidate peginesatide was synthesised as a model compound for this new class of emerging drugs and characterised by means of gel electrophoresis, matrix-assisted laser desorption/ionisation (MALDI) mass spectrometry, as well as liquid chromatography/electrospray ionisation tandem mass spectrometry (LC/ESI-MS/MS) after proteolytic digestion. Based on these results, a mass spectrometric detection method of the product in plasma was developed targeting a pentapeptide fragment after protein precipitation and subtilisin digestion. Its fitness for purpose was evaluated by the determination of selected method characteristics focusing particularly on specificity, recovery (ca. 60%), and limit of detection (1 ng/mL). Verf.-Referat