Scarabelli Silvia, Tan Kui Thong, Griss Rudolf, Hovius Ruud, D'Alessandro Pier Luca, Vorherr Thomas, Johnsson Kai
École Polytechnique Fédérale de Lausanne , Institute of Chemical Sciences and Engineering, Lausanne CH-1015, Switzerland.
National Centre of Competence in Research in Chemical Biology , Lausanne, CH-1015, Switzerland.
ACS Sens. 2017 Aug 25;2(8):1191-1197. doi: 10.1021/acssensors.7b00331. Epub 2017 Aug 2.
We are introducing a new approach to evaluate cellular uptake of drugs and drug candidates into living cells. The approach is based on converting the protein target of a given class of compounds into a fluorescent biosensor. By measuring the binding of different compounds to their cognate biosensor in live cells and comparing these values to those measured in vitro, their cellular uptake and concentrations can be ranked. We demonstrate that our strategy enables the evaluation of the cellular uptake into the cytosol of 2 classes of inhibitors using two different sensor designs; first, sensors comprising the self-labeling protein SNAP conjugated with a chemically modified inhibitor shown for inhibitors of the enzyme human carbonic anhydrase II; and a label-free sensor for inhibitors of protein-protein interactions demonstrated for the protein pair p53-HDM2.
我们正在引入一种新方法来评估药物及候选药物进入活细胞的细胞摄取情况。该方法基于将特定类别的化合物的蛋白质靶点转化为荧光生物传感器。通过测量不同化合物与活细胞中其同源生物传感器的结合情况，并将这些值与体外测量的值进行比较，就可以对它们的细胞摄取和浓度进行排序。我们证明，我们的策略能够使用两种不同的传感器设计来评估两类抑制剂进入细胞质的细胞摄取情况；第一种是包含与化学修饰抑制剂偶联的自标记蛋白SNAP的传感器，用于人碳酸酐酶II抑制剂；第二种是用于蛋白质-蛋白质相互作用抑制剂的无标记传感器，以p53-HDM2蛋白对为例进行了展示。
