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 H460R231Q 细胞中的抗增殖测定和蛋白质印迹分析，鉴定了一系列含腺苷衍生物具有 DOT1LR231Q 抑制作用。最有前途的化合物37可显着降低DOT1LR231Q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。 37 的细胞渗透性和细胞靶标结合通过 CETSA 和 DARTS 测定进行了验证。在 H460R231Q OE 细胞来源的异种移植（CDX）模型中，37 例以 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。一项药代动力学研究表明，37 在大鼠腹膜内给药后具有可耐受的特性（t 1/2 = 1.93 ± 0.91 h，F = 97.2%）。机制研究证实，37种通过MAPK/ERK信号通路抑制携带R231Q功能获得性突变的肺癌的恶性表型。此外，对分子与 DOT1LWT/R231Q 蛋白之间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。© 2024 作者。

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 DOT1LR231Q inhibition through antiproliferation assay and Western blot analysis in the H460R231Q cell. The most promising compound 37 significantly reduced DOT1LR231Q 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 37 were verified by both CETSA and DARTS assays. In the H460R231Q OE cell-derived xenograft (CDX) model, 37 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 37 possessed tolerable properties (t 1/2 = 1.93 ± 0.91 h, F = 97.2%) after intraperitoneal administration in rats. Mechanism study confirmed that 37 suppressed malignant phenotypes of lung cancer carrying R231Q gain-of-function mutation via the MAPK/ERK signaling pathway. Moreover, analysis of the binding modes between molecules and DOT1LWT/R231Q proteins put forward the "Induced-fit" allosteric model in favor to the discovery of potent DOT1L candidates.© 2024 The Authors.