KCNT1基因的功能增益突变,编码钠激活钾通道SLACK,与罕见但破坏性的发育性脑病和癫痫性脑病有关,称为婴儿期癫痫伴迁移局灶性癫痫(EIMFS)。SLACK通道的小分子抑制剂的设计代表了治疗EIMFS的潜在治疗方法,其他儿童癫痫症,和发育障碍。在这里,我们描述了以通过高通量筛选发现的黄嘌呤SLACK抑制剂(8)为中心的命中优化努力。在化学型的三个不同区域,我们合成了58种新的类似物,并在全细胞自动化膜片钳试验中对每种类似物进行了测试,以建立抑制SLACK通道的构效关系.我们进一步评估了所选择的类似物对各种其他离子通道的选择性以及对临床相关SLACK突变体的活性。系列中的选择性相当不错,包括对hERG。类似物80(VU0948578)是WT的有效抑制剂,A934T,和G288S松弛,这些变体的IC50值在0.59和0.71µM之间。VU0948578代表来自化学型的有用的体外工具化合物,其不同于先前报道的SLACK通道的小分子抑制剂。
Gain-of-function mutations in the KCNT1 gene, which encodes the sodium-activated potassium channel known as SLACK, are associated with the rare but devastating developmental and epileptic encephalopathy known as epilepsy of infancy with migrating focal seizures (
EIMFS). The design of small molecule inhibitors of SLACK channels represents a potential therapeutic approach to the treatment of
EIMFS, other childhood epilepsies, and developmental disorders. Herein, we describe a hit optimization effort centered on a xanthine SLACK inhibitor (8) discovered via a high-throughput screen. Across three distinct regions of the chemotype, we synthesized 58 new analogs and tested each one in a whole-cell automated patch-clamp assay to develop structure-activity relationships for inhibition of SLACK channels. We further evaluated selected analogs for their selectivity versus a variety of other ion channels and for their activity versus clinically relevant SLACK mutants. Selectivity within the series was quite good, including versus hERG. Analog 80 (VU0948578) was a potent inhibitor of WT, A934T, and G288S SLACK, with IC50 values between 0.59 and 0.71 µM across these variants. VU0948578 represents a useful in vitro tool compound from a chemotype that is distinct from previously reported small molecule inhibitors of SLACK channels.