需要有效和高度选择性的小分子抑制剂来解开组蛋白甲基转移酶的生物学复杂性并揭示其治疗潜力。开发这些抑制剂的先决条件是鉴定经过验证的化学物质以启动药物化学活动。在大多数情况下,通过筛选大的,公正的化合物库。这些库的大小和性质,再加上利用组蛋白甲基转移酶的双底物的复杂性,需要仔细考虑主屏幕和随后的命中分类。在这一章中,以EZH2为例,我们描述了一项筛查和命中分诊活动,该活动确定了经过验证的化学物质,从而可以启动药物化学研究.此外,我们讨论了一种基于细胞的检测方法,以支持铅的识别和优化。这里描述的方法需要生化的混合物,生物物理和基于细胞的测定应适用于确定其他组蛋白甲基转移酶的验证起点.
Potent and highly selective small-molecule inhibitors are needed to unravel the biological complexities of histone methyltransferases and to reveal their therapeutic potential. A prerequisite to developing these inhibitors is the identification of validated chemical matter for initiating a medicinal chemistry campaign. For the most part, finding these initial starting points occurs through screening of large, unbiased compound libraries. The size and nature of these libraries, coupled with the complexities of the bisubstrate utilizing histone methyltransferases, necessitates that the primary screen and subsequent hit triage be carefully considered.In this chapter, using EZH2 as a representative example, we describe a screening and hit triage campaign that identified validated chemical matter allowing initiation of medicinal chemistry studies. Moreover, we discuss a cell-based assay to support lead identification and optimization. The approach described here entailing a mixture of biochemical, biophysical and cell-based assays should be applicable to identifying validated starting points for other histone methyltransferases.