Atherosclerosis (AS) remains a major global health concern, contributing significantly to cardiovascular morbidity and mortality. Recent research into enzymes like aspartyl aminopeptidase (DNPEP) has shed light on its potential role in regulating inflammation and protein metabolism, offering new avenues for targeted diagnostics and therapies. In light of these challenges, this study aims to develop a novel optical imaging probe (HD-DNPEP) specifically designed to detect DNPEP expression in atherosclerotic lesions. By addressing the current limitations in sensitivity, specificity, and in vivo detection, we seek to create a highly selective and sensitive imaging tool that can provide real-time, high-resolution insights into the role of DNPEP in atherosclerosis. This innovative approach has the potential in the diagnosis of atherosclerosis, enabling more precise intervention strategies and improving patient outcomes. In AS patients, the significantly higher DNPEP levels in the cerebral vasculature and CSF of AS patients suggest that DNPEP may serve as an important biomarker for detecting cerebrovascular arteriosclerosis.