Kapoor Tarun M, Miller Rand M
Laboratory of Chemistry and Cell Biology, The Rockefeller University, 1200 York Ave., New York, NY 10065, USA.
Laboratory of Chemistry and Cell Biology, The Rockefeller University, 1200 York Ave., New York, NY 10065, USA.
Trends Pharmacol Sci. 2017 Dec;38(12):1100-1109. doi: 10.1016/j.tips.2017.09.003. Epub 2017 Oct 13.
Identifying the direct physiological targets of drugs and chemical probes remains challenging. Here we describe how resistance can be used to achieve 'gold-standard' validation of a chemical inhibitor's direct target in human cells. This involves demonstrating that a silent mutation in the target that suppresses inhibitor activity in cell-based assays can also reduce inhibitor potency in biochemical assays. Further, phenotypes due to target inhibition can be identified as those observed in the inhibitor-sensitive cells, across a range of inhibitor concentrations, but not in genetically matched cells with a silent resistance-conferring mutation in the target. We propose that chemotype-specific resistance, which is generally considered to be a limitation of molecularly targeted agents, can be leveraged to deconvolve the mechanism of action of drugs and to properly use chemical probes.
确定药物和化学探针的直接生理靶点仍然具有挑战性。在此，我们描述了如何利用抗性来实现化学抑制剂在人类细胞中直接靶点的“金标准”验证。这涉及证明靶点中的沉默突变在基于细胞的实验中抑制抑制剂活性，同时在生化实验中也能降低抑制剂效力。此外，由于靶点抑制导致的表型可被识别为在一系列抑制剂浓度下在抑制剂敏感细胞中观察到的表型，但在靶点具有沉默抗性赋予突变的基因匹配细胞中则未观察到。我们提出，通常被认为是分子靶向药物局限性的化学型特异性抗性，可被用于解析药物的作用机制并正确使用化学探针。
