基于黄酮的长波长荧光探针用...体外和体内生物硫醇（英文）_周思仪.pdf

有机化学有机化学 Chinese Journal of Organic Chemistry ARTICLE *Corresponding authors.E-mail:; Received June 11,2022;revised August 10,2022;published online September 15,2022.Project supported by the Key Science and Technology Program of Henan Province(Nos.212102311030,222102310671)and the Foundation of Henan Edu-cational Committee(No.22A350012).河南省科技攻关(Nos.212102311030,222102310671)和河南省教育厅基金(No.22A350012)资助项目.178 http:/sioc- Shanghai Institute of Organic Chemistry,Chinese Academy of Sciences Chin.J.Org.Chem.2023,43,178185 DOI:10.6023/cjoc202206016 研究论文研究论文 基于黄酮的长波长荧光探针用于检测体外和体内生物硫醇 周思仪a 丁 旭a 赵永梅b 李景华*,a 罗 稳*,a(a河南大学 天然药物与免疫工程重点实验室 河南开封 475004)(b河南应用技术职业学院 制药工程学院 河南开封 475004)摘要摘要 半胱氨酸(Cys)、同型半胱氨酸(Hcy)和谷胱甘肽(GSH)等生物硫醇在生理和病理过程中起着至关重要的作用,尤其是 GSH 在肿瘤细胞中过度表达,可以作为肿瘤的生物标志物.然而,同时区分活的正常细胞和肿瘤细胞存在着困难和挑战,因此设计并合成了一种基于 N,N-二甲氨基萘黄酮(3)的肉眼可见“关-开”型长波长荧光探针(4).该探针对生物硫醇具有较高的选择性和较低的检测限,机理研究表明,生物硫醇催化探针的酯键断裂,生成了强荧光的黄酮 3.该探针可用于活细胞和小鼠体内生物硫醇成像,并且对正常肝细胞 HL-7702 和肝癌细胞 HepG2 进行选择性成像.关键词关键词 长波长;荧光探针;黄酮;生物硫醇 A Flavone-Based Long-Wavelength Fluorescent Probe to Detect Biothiols in vitro and in vivo Zhou,Siyia Ding,Xua Zhao,Yongmeib Li,Jinghua*,a Luo,Wen*,a(a Key Laboratory of Natural Medicine and Immuno-Engineering,Henan University,Kaifeng,Henan 475004)(b Pharmaceutical Engineering Department,Henan Technical Institute,Kaifeng,Henan 475004)Abstract Biothiols,such as cysteine(Cys),homocysteine(Hcy),and glutathione(GSH)play crucial roles in physiological and pathological processes.GSH,especially,is overexpressed in numerous types of tumor cells and could be considered as a biomarker.However,owing to the difficulty and challenge in sensing normal and cancer cells simultaneously,it is important to solve this problem in biological processes.Herein,a naked-eye visible,“off-on”long-wavelength fluorescent probe(4)based on N,N-(dimethylamino)naphthalenyl flavone(3)was designed and synthesized.This probe showed high selectivity for biothi-ols and low detection limits.The mechanism study revealed that biothiols catalyzed the cleavage of the ester bond,and strongly fluorescent flavone 3 was released.Probe 4 could be used for the imaging of biothiols in living cells and mice.In addition,the probe was optimized to achieve the selective imaging of normal hepatocyte HL-7702 and heptocarcinoma cell HepG2.Keywords long-wavelength;fluorescence probe;flavone;biothiol 1 Introduction Biological thiols(biothiols),including cysteine(Cys),homocysteine(Hcy),and glutathione(GSH),play vital rules in several biological systems.1 For example,a defi-ciency of Cys is associated with albinism,edema,and liver damage.2 Aberrant levels of Hcy are found in cardiovas-cular disease and Alzheimers disease.3 GSH,the most important antioxidant and antidote in mammalian cells,is closely related to the occurrence and development of vari-ous diseases.Pronounced changes in GSH levels affect the intracellular redox homeostasis.4-5 In addition,higher lev-els of GSH have always been found in several cancer cells compared to normal cells.Therefore,GSH can be consid-ered as a biomarker to differentiate between cancerous and normal cell.6-7 The detection and imaging of the cancer biomarker,GSH,may greatly improve the diagnosis of patients with cancer in the early stages and contribute to precision cancer therapy.Hence,efficient methods to de-tect GSH in bio-system,especially for cancer diagnosis,are much needed.In recent years,many methods for biothiol detection have been established,which include electrochemical de-tection and chromatography.8-9 Among existing methods,fluorescence detection is one of the most efficient methods Chinese Journal of Organic Chemistry ARTICLE Chin.J.Org.Chem.2023,43,178185 2023 Shanghai Institute of Organic Chemistry,Chinese Academy of Sciences http:/sioc- owing to its simplicity and non-invasiveness.It is also used for real-time in situ analysis in living systems.10-12 Con-sidering the advantages of the fluorescence method of de-tection,several fluorescent probes based on traditional dyes,such as coumarin,13 naphthalimide,14 rhodamine,15 have been developed,which are widely used for the detec-tion of biothiol in cells and cellular sub-organelles.Compared with other fluorescent dyes,long-wavelength dyes(em600900 nm)have several merits as bioim-aging agents.Long-wavelength dyes have been used as probe for deep tissue penetration,as they cause minimum photo-damage to biological samples and also contribute minimally to background auto-fluorescence interference during the analysis of biomolecules in living systems.16 Therefore,in recent years,numerous long-wavelength flu-orescent probes based on cyanine,17 naphthalene poly-ene,18 and dicyanoisophorone pharmacophores,19 have been developed and used for imaging live systems.How-ever,many of the long-wavelength probes reported are based on the known traditional fluorescent dyes by linking various recognition groups.Novel fluorophore scaffolds have seldom been studied.Flavone,a class of natural products with 2-aryl chro-mone skel