Sainas Stefano, Martino Elena, Garino Claudio, Circosta Paola, Luganini Anna, Giorgis Marta, Bavo Francesco, Piccinini Marco, Ramondetti Cristina, Alberti Marta, Miggiano Riccardo, Gribaudo Giorgio, Boschi Donatella, Frølund Bente, Lolli Marco Lucio
Department of Drug Science and Technology. University of Torino, Via Pietro Giuria 9, Torino 10125, Italy.
Department of Chemistry, University of Torino, Via Pietro Giuria 7, Torino 10125, Italy.
J Med Chem. 2025 Jul 10;68(13):13562-13590. doi: 10.1021/acs.jmedchem.5c00348. Epub 2025 Jun 18.
Fluorescent probes targeting proteins are used to investigate biological processes, requiring strong binding affinity and favorable fluorescence. In this study, we present the first with optimized fluorescence properties. We started exploring the fluorescence of acidic pyrazolo[1,5-]pyridin-2-ol, and, by the introduction of EWGs, π-conjugation, incorporation of push-pull systems and rigid structures, we optimized emission profiles and QY, providing a first (SFR) of the system. To provide proof of concept in biological applications, the established SFR was integrated with DHODH, an important oncology target, enabling the SAR designing fluorescent DHODH and , with being the most potent IC = 170 nM. These inhibitors were validated in vitro for their antileukemic and antiviral activity. As they are both environmentally sensitive fluorescent probes that can highlight their binding to the target, their fluorescence was found to colocalize in the mitochondria, where DHODH is located, in cellular experiments.
靶向蛋白质的荧光探针用于研究生物过程，这需要强大的结合亲和力和良好的荧光特性。在本研究中，我们首次展示了具有优化荧光特性的[具体内容未给出]。我们开始探索酸性吡唑并[1,5 - ]吡啶 - 2 - 醇的荧光，并通过引入电子吸电子基团（EWGs）、π共轭、引入推拉体系和刚性结构，优化了发射光谱和量子产率（QY），提供了该体系的首个[具体内容未给出]（SFR）。为了在生物应用中提供概念验证，将已建立的SFR与重要的肿瘤学靶点二氢乳清酸脱氢酶（DHODH）整合，实现了基于结构活性关系（SAR）设计荧光DHODH抑制剂[具体名称未给出]和[具体名称未给出]，其中[具体名称未给出]是最有效的，其半数抑制浓度（IC）= 170 nM。这些抑制剂在体外针对其抗白血病和抗病毒活性进行了验证。由于它们都是对环境敏感的荧光探针，能够突出显示它们与靶点的结合，在细胞实验中发现它们的荧光共定位于DHODH所在的线粒体中。
