Baker Lisa M, Aimon Anthony, Murray James B, Surgenor Allan E, Matassova Natalia, Roughley Stephen D, Collins Patrick M, Krojer Tobias, von Delft Frank, Hubbard Roderick E
Vernalis (R&D) Ltd, Granta Park, Cambridge, CB21 6GB, UK.
Diamond Light Source Ltd., Harwell Science and Innovation Campus, Didcot, OX11 0QX, UK.
Commun Chem. 2020 Sep 2;3(1):122. doi: 10.1038/s42004-020-00367-0.
Fragment based methods are now widely used to identify starting points in drug discovery and generation of tools for chemical biology. A significant challenge is optimization of these weak binding fragments to hit and lead compounds. We have developed an approach where individual reaction mixtures of analogues of hits can be evaluated without purification of the product. Here, we describe experiments to optimise the processes and then assess such mixtures in the high throughput crystal structure determination facility, XChem. Diffraction data for crystals of the proteins Hsp90 and PDHK2 soaked individually with 83 crude reaction mixtures are analysed manually or with the automated XChem procedures. The results of structural analysis are compared with binding measurements from other biophysical techniques. This approach can transform early hit to lead optimisation and the lessons learnt from this study provide a protocol that can be used by the community.
基于片段的方法目前广泛应用于药物发现的起始点识别以及化学生物学工具的生成。一个重大挑战是将这些弱结合片段优化为活性化合物和先导化合物。我们开发了一种方法，无需纯化产物即可评估命中类似物的各个反应混合物。在此，我们描述了优化这些过程的实验，然后在高通量晶体结构测定设施XChem中评估此类混合物。分别用83种粗反应混合物浸泡的热休克蛋白90（Hsp90）和丙酮酸脱氢酶激酶2（PDHK2）晶体的衍射数据通过手动或XChem自动化程序进行分析。结构分析结果与其他生物物理技术的结合测量结果进行比较。这种方法可以将早期的活性化合物转化为先导化合物优化，并且从本研究中吸取的经验教训提供了一个可供同行使用的方案。
