Citation: Zhihui Zhang, Ru Sun, Chong Bian, Hongbo Wang, Zhen Zhao, Panpan Lv, Jianzhong Lu, Haixin Zhang, Hulie Zeng, Yuanyuan Chen, Zhijuan Cao. A dual-protease-triggered chemiluminescent probe for precise tumor imaging[J]. Chinese Chemical Letters, ;2025, 36(2): 109784. doi: 10.1016/j.cclet.2024.109784 shu

A dual-protease-triggered chemiluminescent probe for precise tumor imaging

Zhihui Zhang ^{a, 1} ,
Ru Sun ^{a, 1} ,
Chong Bian ^{b, 1} ,
Hongbo Wang ^c ,
Zhen Zhao ^b ,
Panpan Lv ^b ,
Jianzhong Lu ^a ,
Haixin Zhang ^a ,
Hulie Zeng ^a ,
Yuanyuan Chen ^d ,
Zhijuan Cao ^{a, b, *,}

a.
Department of Pharmaceutical Analysis, School of Pharmacy, Fudan University, Shanghai 201203, China
b.
Minhang Branch, Zhongshan Hospital, Fudan University, Shanghai 201199, China
c.
China State Institute of Pharmaceutical Industry, Shanghai 200040, China
d.
Shanghai Jahwa United Co., Ltd., Shanghai 200082, China

zjcao@fudan.edu.cn

Received Date: 15 December 2023
Revised Date: 9 March 2024
Accepted Date: 15 March 2024
Available Online: 22 March 2024

Figures(6)

Imaging detection of interlinked dual proteases is imperative for precise tumor imaging, which remains challenging due to limited modification position of specific substrate and possible steric hindrance. Herein, we have developed a unimolecular chemiluminescent probe (LGP-CL) tandemly activated by two proteases interlinked with liver cancer to achieve precise tumor imaging. Probe LGP-CL consists of a phenoxy-dioxetane scaffold caged by a tripeptide substrate (LGP, leucine-glycine-proline) as the sensing layer, which can be cleaved sequentially by aminopeptidase N (APN) and dipeptidyl peptidase Ⅳ (DPPIV) to turn on a strong chemiluminescent signal, and silenced by specific inhibitor of each enzyme, which accounts for an integrated logic gate (AND, OR and INHIBIT). The successful cleavage of dual proteases on the metabolic site depends on the proper structure of the tripeptide substrate, as confirmed by two probes design. Probe LGP-CL (LGP as the substrate) enables the excellent "dual-lock-dual-key" fit with a 382-fold enhancement of chemiluminescent emission while no obvious signal is observed by using GPL-CL (GPL as the substrate). By virtue of its rapid response (several minutes), high sensitivity and good cell viability, probe LGP-CL has been utilized to evaluate upregulated levels of proteases in vitro and in living systems, especially to distinguish liver tumor cells (HepG2) from others (LO2, MCF-7, MCF-10a and RAW264.7). Overall, the newly developed CL probe may facilitate rapid investigation into the role played by proteases in liver diseases, enabling timely selection appropriate treatment. Therefore, our work not only sheds light on the rational design of optical probes for dual protease imaging, but provides a promising tool for clinical diagnosis and even drug discovery.
- Dual-protease imaging,
- Tripeptide substrate,
- Chemiluminescent,
- Phenoxy-dioxetane scaffold,
- Logic gates
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