Tau tubulin kinase 1 and 2 (TTBK1/2) are highly homologous kinases that are expressed and mediate disease-relevant pathways predominantly in the brain. Distinct roles for TTBK1 and TTBK2 have been delineated. While efforts have been devoted to characterizing the impact of TTBK1 inhibition in diseases like Alzheimer’s disease and amyotrophic lateral sclerosis, TTBK2 inhibition has been less explored. TTBK2 serves a critical function during cilia assembly. Given the biological importance of these kinases, we designed a targeted library from which we identified several chemical tools that engage TTBK1 and TTBK2 in cells and inhibit their downstream signaling. Indolyl pyrimidinamine 10 significantly reduced the expression of primary cilia on the surface of human induced pluripotent stem cells (iPSCs). Furthermore, analog 10 phenocopies TTBK2 knockout in iPSCs, confirming a role for TTBK2 in ciliogenesis.

Tau 微管蛋白激酶 1 和 2 (TTBK1/2) 是高度同源的激酶，主要在大脑中表达并介导疾病相关通路。 TTBK1 和 TTBK2 的不同作用已被描述。虽然人们一直致力于表征 TTBK1 抑制对阿尔茨海默病和肌萎缩侧索硬化症等疾病的影响，但对 TTBK2 抑制的研究较少。 TTBK2 在纤毛组装过程中发挥着关键作用。考虑到这些激酶的生物学重要性，我们设计了一个靶向文库，从中鉴定了几种与细胞中的 TTBK1 和 TTBK2 结合并抑制其下游信号传导的化学工具。吲哚基嘧啶胺10显着降低人诱导多能干细胞 (iPSC) 表面初级纤毛的表达。此外，类似物10在 iPSC 中表现出 TTBK2 敲除的表型，证实了 TTBK2 在纤毛发生中的作用。