Aberrant activation of casein kinase 1 (CK1) has been demonstrated to be implicated in the pathogenesis of cancer and various central nervous system disorders. Discovery of CK1 inhibitors has thus attracted much attention in recent years. In this account, we describe the discovery of N6-phenyl-1H-pyrazolo[3,4-d]pyrimidine-3,6-diamine derivatives as novel CK1 inhibitors. An optimal common-feature pharmacophore hypothesis, termed Hypo2, was firstly generated, followed by virtual screening using Hypo2 against several chemical databases. One of the best hit compounds, N6-(4-chlorophenyl)-1H-pyrazolo[3,4-d]pyrimidine-3,6-diamine, was chosen for the subsequent hit-to-lead optimization under the guide of Hypo2, which led to the discovery of a new lead compound (1-(3-(3-amino-1H-pyrazolo[3,4-d]pyrimidin-6-ylamino)phenyl)-3-(3-chloro-4-fluorophenyl)urea) that potently inhibits CK1 with an IC₅₀ value of 78 nM.

酪蛋白激酶1（CK1）的异常激活已被证明与癌症和各种中枢神经系统疾病的发病机制有关。因此，近年来CK1抑制剂的发现引起了很多关注。在此说明中，我们描述了作为新型CK1抑制剂的N6-苯基-1 H-吡唑并[3,4-d]嘧啶-3,6-二胺衍生物的发现。首先生成了最佳的共同特征药效基团假说，称为Hypo2，然后使用Hypo2对几种化学数据库进行虚拟筛选。选择了最佳命中化合物之一，即N6-（4-氯苯基）-1 H-吡唑并[3,4-d]嘧啶-3,6-二胺，用于随后在Hypo2指导下进行命中优化。 ，导致发现了新的先导化合物（1-（3-（3-amino-1 H-吡唑并[3,4-d]嘧啶-6-基氨基）苯基）-3-（3-氯-4-氟苯基）脲，可有效抑制CK1，IC ₅₀值为78 nM。