Drug Screening in Human Cells by NMR Spectroscopy Allows the Early Assessment of Drug Potency
Drug Screening in Human Cells by NMR Spectroscopy Allows the Early Assessment of Drug Potency
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DOI：
10.1002/anie.201913436
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发表时间：
2020-02-25
影响因子：
16.6
通讯作者：
Banci, Lucia
中科院分区：
文献类型：
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作者：
Luchinat, Enrico;Barbieri, Letizia;Banci, Lucia
基于结构的药物开发通常受到体外筛选的有希望的化合物缺乏体内活性的阻碍，这是由于低的膜渗透性或差的细胞内结合选择性。在此，我们表明，配体筛选可以在活的人类细胞中进行“细胞内蛋白质观察”NMR光谱，而不需要酶活性测量或其他细胞测定。通过快速、廉价的H-1 NMR实验获得定量结合信息，提供细胞内剂量和时间依赖性配体结合曲线，从中获得与细胞渗透性和结合亲和力和选择性相关的动力学和热力学参数。该方法适用于碳酸酐酶，原则上，可以扩展到任何NMR可观察到的细胞内目标。所获得的结果与候选药物的效力，即所需剂量直接相关。在药物设计管道的早期阶段应用这种方法可以大大提高现代药物开发的低成功率。
Structure-based drug development is often hampered by the lack of in vivo activity of promising compounds screened in vitro, due to low membrane permeability or poor intracellular binding selectivity. Herein, we show that ligand screening can be performed in living human cells by "intracellular protein-observed" NMR spectroscopy, without requiring enzymatic activity measurements or other cellular assays. Quantitative binding information is obtained by fast, inexpensive H-1 NMR experiments, providing intracellular dose- and time-dependent ligand binding curves, from which kinetic and thermodynamic parameters linked to cell permeability and binding affinity and selectivity are obtained. The approach was applied to carbonic anhydrase and, in principle, can be extended to any NMR-observable intracellular target. The results obtained are directly related to the potency of candidate drugs, that is, the required dose. The application of this approach at an early stage of the drug design pipeline could greatly increase the low success rate of modern drug development.