Center for the Science of Therapeutics, Broad Institute, Cambridge, MA, 02142, USA.
Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, 02215, USA.
ChemMedChem. 2016 Dec 6;11(23):2575-2581. doi: 10.1002/cmdc.201600502. Epub 2016 Nov 15.
Evaluating the engagement of a small molecule ligand with a protein target in cells provides useful information for chemical probe optimization and pharmaceutical development. While several techniques exist that can be performed in a low-throughput manner, systematic evaluation of large compound libraries remains a challenge. In-cell engagement measurements are especially useful when evaluating compound classes suspected to target multiple cellular factors. In this study we used a bioluminescent resonant energy transfer assay to assess bromodomain engagement by a compound series containing bromodomain- and kinase-biasing polypharmacophores based on the known dual BRD4 bromodomain/PLK1 kinase inhibitor BI2536. With this assay, we discovered several novel agents with bromodomain-selective specificity profiles and cellular activity. Thus, this platform aids in distinguishing molecules whose cellular activity is difficult to assess due to polypharmacologic effects.
评估小分子配体与细胞内蛋白质靶标的结合情况，可为化学探针优化和药物开发提供有用信息。虽然有几种技术可以以低通量方式进行，但对大型化合物库进行系统评估仍然是一个挑战。当评估疑似靶向多种细胞因子的化合物类别时，细胞内结合测量特别有用。在这项研究中，我们使用生物发光共振能量转移测定法来评估一系列包含基于已知双重 BRD4 溴结构域/PLK1 激酶抑制剂 BI2536 的溴结构域和激酶偏向多药效团的化合物对溴结构域的结合情况。通过该测定法，我们发现了几种具有溴结构域选择性特异性谱和细胞活性的新型试剂。因此，该平台有助于区分由于多药效学作用而难以评估细胞活性的分子。
