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...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 a382-fold enhancement of chemiluminescent emission while no obvious signal is observed by using GPLCL(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(Hep G2)from others(LO_(2),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.展开更多
基金financial support by National Key R&D Program of China,MOST(No.2023YFC2510000)Shanghai Science and Technology(No.21N31900500)+2 种基金Shanghai Municipal Health Commission Project(No.202140016)Training Program for Outstanding Young Medical and Pharmaceutical Talents of Minhang District Health System(No.mwyjyx08)the Project of Basic Medicine funded by Fudan-Minhang Health Consortium(Nos.2021MHJC10 and 2023FM09)。
文摘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 a382-fold enhancement of chemiluminescent emission while no obvious signal is observed by using GPLCL(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(Hep G2)from others(LO_(2),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.
