Garbagnoli Martina, Linciano Pasquale, Listro Roberta, Rossino Giacomo, Vasile Francesca, Collina Simona
Department of Drug Sciences, University of Pavia, viale Taramelli 12, Pavia 27100, Italy.
Department of Chemistry, University of Milan, Via Golgi 19, Milano 20133, Italy.
ACS Omega. 2024 Apr 10;9(16):17691-17705. doi: 10.1021/acsomega.4c01309. eCollection 2024 Apr 23.
Drug discovery is a lengthy and intricate process, and in its early stage, crucial steps are the selection of the therapeutic target and the identification of novel ligands. Most targets are dysregulated in pathogenic cells; typically, their activation or deactivation leads to the desired effect, while in other cases, interfering with the target-natural binder complex achieves the therapeutic results. Biophysical assays are a suitable strategy for finding new ligands or interferent agents, being able to evaluate ligand-protein interactions and assessing the effect of small molecules (SMols) on macromolecular complexes. This mini-review provides a detailed analysis of widely used biophysical methods, including fluorescence-based approaches, circular dichroism, isothermal titration calorimetry, microscale thermophoresis, and NMR spectroscopy. After a brief description of the methodologies, examples of interaction and competition experiments are described, together with an analysis of the advantages and disadvantages of each technique. This mini-review provides an overview of the most relevant biophysical technologies that can help in identifying SMols able not only to bind proteins but also to interfere with macromolecular complexes.
药物发现是一个漫长而复杂的过程，在其早期阶段，关键步骤是治疗靶点的选择和新型配体的鉴定。大多数靶点在致病细胞中表达失调；通常，它们的激活或失活会产生预期效果，而在其他情况下，干扰靶点-天然结合物复合物可实现治疗效果。生物物理测定是寻找新配体或干扰剂的合适策略，能够评估配体-蛋白质相互作用并评估小分子对大分子复合物的影响。本综述对广泛使用的生物物理方法进行了详细分析，包括基于荧光的方法、圆二色性、等温滴定量热法、微量热泳动和核磁共振光谱法。在简要描述这些方法后，介绍了相互作用和竞争实验的实例，并分析了每种技术的优缺点。本综述概述了最相关的生物物理技术，这些技术有助于鉴定不仅能够结合蛋白质而且能够干扰大分子复合物的小分子。
