Highly Sensitive Chemiluminescent Probe for CYP 2J2 Detection and Image-Guided Liver Cancer Surgery

Developing chemiluminescent (CL) probes with a high tumor-to-normal tissue ratio is crucial for liver tumor visualization and image-guided surgery. Cytochrome P450 2J2 (CYP 2J2), an extrahepatic enzyme, is highly expressed in liver tumors but not in healthy tissues. However, no prior reports have documented the development of CL probes specifically targeting CYP 2J2. In this study, we designed a CYP 2J2-responsive CL probe, termed CYP-PD, by grafting methoxybenzyl alcohol onto Schaap's dioxetanes. In vitro and cellular experiments confirmed that CYP-PD selectively and sensitively responds to both exogenous and endogenous CYP 2J2, exhibiting an extraordinary limit of detection of 1.21 pM and a cellular detection threshold as low as 235 cells. Moreover, CYP-PD demonstrated the highest CL intensity in HepG-2 cells, with an ∼59.6-fold enhancement relative to normal liver cells (L02). Notably, the probe enabled a high tumor-to-normal tissue visualization ratio of tumor lesions from normal liver tissues (∼44.7-fold), successfully guiding the surgical resection of an orthotopic liver cancer mouse model. We envision that this work may provide a powerful tool for CYP 2J2 detection and image-guided liver cancer surgery in the future.