Department of Nuclear Medicine, Peking University First Hospital, Beijing 100034, China.
Department of Nuclear Medicine, Beijing Chaoyang Hospital, Capital Medical University, Beijing 100020, China.
Bioorg Med Chem Lett. 2020 Jul 15;30(14):127253. doi: 10.1016/j.bmcl.2020.127253. Epub 2020 May 11.
To facilitate the discovery of FAP inhibitors, a convenient cell-based fluorescent assay was developed by using a commonly available U87MG cell line and a FAP-specific substrate Suc-Gly-Pro-AMC. The assay enabled the fast determination of multiple ICs by simply incubating a solution of phosphate-buffered saline in a 96-well plate within 30 min. The substrate specificity, cross-reaction and other related conditions were systematically optimized. This method was successfully applied to determine the ICs of seven known inhibitors. The results are in consistence with the trend reported, which indicating that this practical assay is a valuable method to accelerate the discovery of FAP inhibitor.
为了促进 FAP 抑制剂的发现，我们采用一种常见的 U87MG 细胞系和一种 FAP 特异性底物 Suc-Gly-Pro-AMC，开发了一种方便的基于细胞的荧光测定法。该测定法通过在 96 孔板中简单孵育磷酸盐缓冲盐水溶液，在 30 分钟内即可快速测定多个 IC。我们对底物特异性、交叉反应性和其他相关条件进行了系统优化。该方法成功应用于测定七种已知抑制剂的 IC。结果与报道的趋势一致，这表明这种实用的测定法是加速 FAP 抑制剂发现的有价值的方法。
