Recent research certified that DOT1L and its mutations represented by R231Q were potential targets for the treatment of lung cancer. Herein, a series of adenosine-containing derivatives were identified with DOT1L inhibition through antiproliferation assay and Western blot analysis in the H460 cell. The most promising compound significantly reduced DOT1L mediated H3K79 methylation and effectively inhibited the proliferation, self-renewal, migration, and invasion of lung cancer cell lines at low micromolar concentrations. The cell permeability and cellular target engagement of were verified by both CETSA and DARTS assays. In the H460 cell-derived xenograft (CDX) model, displayed pronounced tumor growth inhibition after intraperitoneal administration at 20 mg/kg dose for 3 weeks (TGI = 54.38%), without obvious toxicities. A pharmacokinetic study revealed that possessed tolerable properties ( = 1.93 ± 0.91 h, = 97.2%) after intraperitoneal administration in rats. Mechanism study confirmed that suppressed malignant phenotypes of lung cancer carrying R231Q gain-of-function mutation the MAPK/ERK signaling pathway. Moreover, analysis of the binding modes between molecules and DOT1L proteins put forward the “Induced-fit” allosteric model in favor to the discovery of potent DOT1L candidates.

中文翻译：

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发现针对催化结构域中 R231Q 功能获得性突变的首创 DOT1L 抑制剂，具有肺癌治疗潜力

最近的研究证明，DOT1L及其以R231Q为代表的突变是肺癌治疗的潜在靶点。在此，通过 H460 细胞中的抗增殖测定和蛋白质印迹分析，鉴定了一系列含有腺苷的衍生物具有 DOT1L 抑制作用。最有前途的化合物可显着降低 DOT1L 介导的 H3K79 甲基化，并在低微摩尔浓度下有效抑制肺癌细胞系的增殖、自我更新、迁移和侵袭。通过 CETSA 和 DARTS 测定验证了细胞通透性和细胞靶标结合。在H460细胞来源的异种移植（CDX）模型中，以20 mg/kg剂量腹腔给药3周后表现出明显的肿瘤生长抑制（TGI = 54.38%），且无明显毒性。药代动力学研究表明，大鼠腹膜内给药后具有可耐受的特性（= 1.93 ± 0.91 h，= 97.2%）。机制研究证实，携带R231Q功能获得性突变的MAPK/ERK信号通路可抑制肺癌的恶性表型。此外，对分子与 DOT1L 蛋白之间的结合模式的分析提出了“诱导拟合”变构模型，有利于发现有效的 DOT1L 候选物。