Comparison of ¹²⁵I- and ¹¹¹In-labeled peptide probes for in vivo detection of oxidized low-density lipoprotein in atherosclerotic plaques

Oxidized low-density lipoprotein (OxLDL) plays a pivotal role in atherosclerotic plaque destabilization, which suggests its potential as a nuclear medical imaging target. We previously developed radioiodinated ¹²⁵I-AHP7, a peptide probe carrying a 7-residue sequence from the OxLDL-binding protein Asp-hemolysin, for specific OxLDL imaging. Although ¹²⁵I-AHP7 recognized OxLDL, it had low stability. Thus, to improve stability, we designed radiolabeled 22-residue peptide probes, ¹²⁵I-AHP22 and ¹¹¹In-AHP22, which include the entire AHP7 sequence, and evaluated the stability, activity, and applications of these probes in vitro and in vivo.

Probes consisting of a 21-residue peptide derived from the Asp-hemolysin sequence and an N-terminal Cys or aminohexanoic acid for labeling with ¹²⁵I-N-(3-iodophenyl)maleimide or ¹¹¹In diethylene triamine pentaacetic acid were termed ¹²⁵I-AHP22 and ¹¹¹In-AHP22. An in vitro-binding inhibition assay with OxLDL was performed using ¹²⁵I-AHP7 as a radiotracer. Radioactivity accumulation in the atherosclerotic aorta and plasma intact fraction was evaluated 30 min after intravenous administration of probes in myocardial infarction-prone Watanabe heritable hyperlipidemic (WHHLMI) rabbits.

¹²⁵I-AHP22 and ¹¹¹In-AHP22 were synthesized in ~ 360 and 60 min, respectively, with > 98% radiochemical purities after RP-HPLC purification. An in vitro-binding assay revealed similar or greater inhibition of OxLDL binding by both In-AHP22 and I-AHP22 compared to I-AHP7. The fraction of intact ¹²⁵I-AHP22 and ¹¹¹In-AHP22 in plasma was estimated to be approximately tenfold higher than that of ¹²⁵I-AHP7. Both probes were rapidly cleared from the blood. ¹¹¹In-AHP22 had a 2.3-fold higher accumulation in WHHLMI rabbit aortas compared to control rabbits, which was similar to ¹²⁵I-AHP7. However, ¹²⁵I-AHP22 accumulated to similar levels in aortas of WHHLMI and control rabbits due to high nonspecific accumulation in normal aortas that could be due to high lipophilicity.

¹¹¹In-AHP22, easily prepared within 1 h, showed moderate affinity for OxLDL, high stability in vivo, and high accumulation in atherosclerotic aortas. ¹¹¹In-AHP22 could be a potential lead compound to develop future effective OxLDL imaging probes.