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.

基于片段的方法现在广泛用于确定药物发现和化学生物学工具生成的起点。一个重大挑战是优化这些弱结合片段以命中和先导化合物。我们开发了一种方法，可以在不纯化产品的情况下评估命中类似物的单个反应混合物。在这里，我们描述了优化流程的实验，然后在高通量晶体结构测定设备 XChem 中评估此类混合物。手动或使用自动 XChem 程序分析用 83 种粗反应混合物单独浸泡的蛋白质 Hsp90 和 PDHK2 晶体的衍射数据。将结构分析的结果与其他生物物理技术的结合测量结果进行比较。