A dual-locked hepatic-targeting fluorescent probe for hepatocellular carcinoma imaging with HClO and β-galactosidase activation

Hepatocellular carcinoma (HCC) is one of the most common types of cancers. Identifying and validating specific and robust cancer biomarkers is paramount for the early screening, timely diagnosis, and accurate prognosis of a cancer. Overexpression of hypochlorous acid (HClO) and β -galactosidase ( β -gal) in malignant tumors is considered a key biomarker of cancer pathology. We herein present a groundbreaking approach: the design and synthesis of a tandem dual-locked fluorescent probe, named LDMTY. Compared to conventional single-locked probes, LDMTY demonstrates superior potential for enhancing the diagnostic specificity of HCC. LDMTY exhibits excellent sensitivity and specificity toward HClO and β -gal, with minimum detection limits of 0.046 μM and 2.54 × 10⁻⁴ U/mL, respectively. Because of the reaction between LDMTY and overexpressed β -gal and HClO present with in cancerous cells, the fluorescence of LDMTY significantly increases (by 220-fold), enabling precise differentiation of between cancerous cells from normal cells ( P < 0.0001). Moreover, the galactose modification on LDMTY enhances its hepatic-targeting capabilities, significantly enhancing its HCC imaging accuracy. LDMTY was successfully applied to in vivo imaging studies in mouse HCC models. It demonstrated promising potential as an innovative tandem dual-locked visual analysis tool and would offer value for HCC-specific detection.