P21-activated kinase 1 (PAK1) is associated with cell proliferation, survival and migration. Deregulation of PAK1 activity is involved in various human diseases, including cancer, inflammation, and neurological disorders. Using a high-throughput virtual screening, we identified the 1H-pyrazolo [3,4-d]pyrimidine scaffold as a promising lead for targeting PAK1. We designed and synthesized a focused library through a structure-based strategy. A novel potent PAK1 inhibitor, ZMF-10, was discovered, which presented an IC₅₀ value of 174 nM with a good selectivity. In addition, ZMF-10 could inhibit PAK1-ERK signaling to suppress MDA-MB-231 cells proliferation with an IC₅₀ value of 3.48 μM for 48 h. Subsequently, ZMF-10 was documented to induce cell apoptosis. Interestingly, according to the RNASeq-based analyses, we substantiated that ZMF-10 induced significant ER-Stress, suppressed migration via FOXO3 activation, JNK1/2, ERK1/2 and AKT signaling inhibition. Together, these results demonstrate that ZMF-10 is a novel PAK1 inhibitor triggering apoptosis, ER stress and anti-migration effect in MDA-MB-231 cells, which may provide a candidate lead for the development of novel potent inhibitors of PAK1.

P21激活的激酶1（PAK1）与细胞增殖，存活和迁移有关。PAK1活性的失调涉及多种人类疾病，包括癌症，炎症和神经系统疾病。使用高通量虚拟筛选，我们确定了1 H-吡唑并[3,4- d ]嘧啶支架是靶向PAK1的有前途的线索。我们通过基于结构的策略设计并综合了一个重点库。发现了一种新型的有效PAK1抑制剂ZMF-10，其IC ₅₀值为174 nM，具有良好的选择性。此外，ZMF-10可以通过IC ₅₀抑制PAK1-ERK信号传导，从而抑制MDA-MB-231细胞增殖。值3.48μM持续48小时。随后，据报道ZMF-10诱导细胞凋亡。有趣的是，根据基于RNASeq的分析，我们证实ZMF-10诱导了显着的ER-应力，通过FOXO3激活，JNK1 / 2，ERK1 / 2和AKT信号传导抑制抑制了迁移。总之，这些结果表明，ZMF-10是一种新型的PAK1抑制剂，可在MDA-MB-231细胞中触发凋亡，内质网应激和抗迁移作用，这可能为开发新型有效的PAK1抑制剂提供了候选线索。