Chia Nan University of Pharmacy & Science Institutional Repository:Item 310902800/27685
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    標題: An Activity-Based Near-Infrared Glucuronide Trapping Probe for Imaging beta-Glucuronidase Expression in Deep Tissues
    作者: Cheng, Ta-Chun
    Roffler, Steve R.
    Tzou, Shey-Cherng
    Chuang, Kuo-Hsiang
    Su, Yu-Cheng
    Chuang, Chih-Hung
    Kao, Chien-Han
    Chen, Chien-Shu
    Harn, I-Hong
    Liu, Kuan-Yi
    Cheng, Tian-Lu
    Leu, Yu-Ling
    貢獻者: 藥學系
    關鍵字: In-Vivo
    Reporter Gene
    Prodrug Monotherapy
    日期: 2012-02-15
    上傳時間: 2014-03-21 16:17:37 (UTC+8)
    出版者: Amer Chemical Soc
    摘要: beta-glucuronidase is an attractive reporter and prodrug-converting enzyme. The development of near-IR (NIR) probes for imaging of beta-glucuronidase activity would be ideal to allow estimation of reporter expression and for personalized glucuronide prodrug cancer therapy in preclinical studies. However, NIR glucuronide probes are not yet available. In this work, we developed two fluorescent probes for detection of beta-glucuronidase activity, one for the NIR range (containing IR-820 dye) and the other for the visible range [containing fluorescein isothiocyanate (FITC)], by utilizing a difluoromethylphenol-glucuronide moiety (TrapG) to trap the fluorochromes in the vicinity of the active enzyme. beta-glucuronidase-mediated hydrolysis of the glucuronyl bond of TrapG generates a highly reactive alkylating group that facilitates the attachment of the fluorochrome to nucleophilic moieties located near beta-glucuronidase-expressing sites. FITC-TrapG was selectively trapped on purified beta-glucuronidase or beta-glucuronidase-expressing CT26 cells (CT26/m beta G) but not on bovine serum albumin or non-beta-glucuronidase-expressing CT26 cells used as controls. beta-glucuronidase-activated FITC-TrapG did not interfere with beta-glucuronidase activity and could label bystander proteins near beta-glucuronidase. Both FITC-TrapG and NIR-TrapG specifically imaged subcutaneous CT26/m beta G tumors, but only NIR-TrapG could image CT26/m beta G tumors transplanted deep in the liver. Thus NIR-TrapG may provide a valuable tool for visualizing beta-glucuronidase activity in vivo.
    显示于类别:[藥學系(所)] 期刊論文


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