The bone marrow tyrosine kinase in chromosome X (BMX) is pursued as a drug target because of its role in various pathophysiological processes. We designed BMX covalent inhibitors with single-digit nanomolar potency with unexploited topological pharmacophore patterns. Importantly, we reveal the first X-ray crystal structure of covalently inhibited BMX at Cys496, which displays key interactions with Lys445, responsible for hampering ATP catalysis and the DFG-out-like motif, typical of an inactive conformation. Molecular dynamic simulations also showed this interaction for two ligand/BMX complexes. Kinome selectivity profiling showed that the most potent compound is the strongest binder, displays intracellular target engagement in BMX-transfected cells with two-digit nanomolar inhibitory potency, and leads to BMX degradation PC3 in cells. The new inhibitors displayed anti-proliferative effects in androgen-receptor positive prostate cancer cells that where further increased when combined with known inhibitors of related signaling pathways, such as PI3K, AKT and Androgen Receptor. We expect these findings to guide development of new selective BMX therapeutic approaches.

中文翻译：

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对合理设计的有效 BMX 抑制剂共价结合模式的结构和生物物理学见解

X 染色体 (BMX) 中的骨髓酪氨酸激酶因其在各种病理生理过程中的作用而被视为药物靶点。我们设计了具有个位数纳摩尔效力的 BMX 共价抑制剂和未开发的拓扑药效团模式。重要的是，我们揭示了 Cys496 处共价抑制 BMX 的第一个 X 射线晶体结构，它显示与 Lys445 的关键相互作用，负责阻碍 ATP 催化和 DFG-out 样基序，典型的非活性构象。分子动力学模拟也显示了两种配体/BMX 复合物的这种相互作用。激酶组选择性分析表明，最有效的化合物是最强的结合剂，在 BMX 转染细胞中显示出细胞内靶标参与，具有两位数的纳摩尔抑制效力，并导致 BMX 降解细胞中的 PC3。新抑制剂在雄激素受体阳性前列腺癌细胞中显示出抗增殖作用，当与已知的相关信号通路抑制剂（如 PI3K、AKT 和雄激素受体）结合时，这种作用会进一步增加。我们希望这些发现能够指导新的选择性 BMX 治疗方法的开发。