Technetium-99 m-PEGylated dendrimer-G2-(Dabcyle-Lys6,Phe7)-pHBSP: A novel Nano-Radiotracer for molecular and early detecting of cardiac ischemic region

Abstract

In cardiac ischemic disorder, pyroglutamate helix B surface peptide (pHBSP) which derived from erythropoietin causes to increase cell stability. To improve the serum stability of pHBSP, two lipophilic amino acids Arg⁶, Ala⁷ were replaced with Fmoc-(Dabcyle)-Lys-OH and Fmoc-Phe-OH during the peptide synthesis. This peptide was subsequently conjugated to PEGylated dendrimer-G₂ and labeled with ^99mTcO₄^- to detect cardiac ischemic region. Radiochemical purity (RCP) of ^99mTc-PEGylated dendrimer-G₂-(Dabcyle-Lys⁶,Phe⁷)-pHBSP was evaluated by ITLC method. In addition, the radiopeptide was investigated for stability in human serum and binding affinity to hypoxic cells in myocardium H9c2 cell lines. Biodistribution and SPECT/CT scintigraphy were assessed in cardiac ischemic rats. Radiochemical yield indicated that the anionic dendrimer has a very high potential to complex formation with ^99mTcO^-₄ (RCP > 94%) which was stable in human serum with RCP 89% up to 6 h. The binding of ^99mTc- nanoconjugate to hypoxic cells was significantly more than normoxic cells (3-fold higher compared to normoxic cells at 1 h). In biodistribution studies, erythropoietin receptor-Beta common receptor (EPO-BcR)-positive uptake in the cardiac ischemic region was 3.62 ± 0.44% ID/g 30 min post injection. SPECT imaging showed a prominent uptake of ^99mTc-nanoconjugate in EPO-BcR expressing ischemic heart.

Keywords: Ischemia; Labeling; PEGylated dendrimer-G(2); Technetium-99m; pHBSP.