Nonerythropoietic erythropoietins (EPOs) are investigated for their high antioxidant properties. A new drug candidate under clinical investigation to treat brain diseases is Neuro-EPO, produced by selecting EPO isoforms with low sialic acid content. Intranasal administration allows to bypass the blood-brain barrier to get a fast and concentrated delivery to the brain. The aims of this project were to characterize Neuro-EPO with anti-doping methods used to detect conventional recombinant EPOs (isoelectric focusing [IEF] and sodium dodecyl sulfate-polyacrylamide gel electrophoresis [SDS-PAGE]) and to evaluate the window of detection of Neuro-EPO in brain and blood (plasma) after a single intranasal administration in rats. Neuro-EPO drug analyzed by IEF-PAGE presented a very basic profile completely detected only when using a 2-8 or 2-10 pH gradient instead of the conventional 2-6 pH gradient. Its profile consisted in six main bands that did not interfere with endogenous EPO profile from human or rat. After SDS-PAGE, a broad band was detected for Neuro-EPO in the same area as endogenous EPO, making Neuro-EPO identification very difficult by this approach. Therefore, IEF was the method for identification chosen after administration in rats. Neuro-EPO was clearly identified in blood 2 and 6 h after the delivery. Fainter signals were obtained between 12 and 48 h, but some characteristic very basic bands remained detectable. Surprisingly, brain extracts did not show the presence of Neuro-EPO even 2 h after administration, indicating a fast degradation or elimination from the brain to the bloodstream. This experiment indicated that detection of Neuro-EPO after intranasal delivery should be possible for a few days.
Keywords: IEF-PAGE; Neuro-EPO; brain; detection; serum.
© 2020 John Wiley & Sons, Ltd.