A non-covalent inhibitor XMU-MP-3 overrides ibrutinib-resistant BtkC481S mutation in B-cell malignancies
A non-covalent inhibitor XMU-MP-3 overrides ibrutinib-resistant BtkC481S mutation in B-cell malignancies
复制标题
非共价抑制剂 XMU-MP-3 可以克服 B 细胞恶性肿瘤中的依鲁替尼耐药 Btk(C481S) 突变
DOI：
10.1111/bph.14809
复制
发表时间：
2019-12-09
影响因子：
7.3
通讯作者：
Deng, Xianming
中科院分区：
文献类型：
--
作者：
Gui, Fu;Jiang, Jie;Deng, Xianming
背景与目的布鲁顿酪氨酸激酶（Bruton's tyrosine kinase， BTK）在多种b细胞恶性肿瘤中通过调节细胞增殖和存活，在b细胞受体信号传导中起关键作用。共价低分子量BTK激酶抑制剂在b细胞恶性肿瘤中显示出令人印象深刻的临床疗效。然而，突变体Btk（C481S）通过破坏Btk与不可逆抑制剂（如伊鲁替尼）之间共价键的形成，对b细胞恶性肿瘤的管理构成了重大挑战。本研究旨在开发针对ibrutinib耐药BTK （C481S）突变的新型BTK抑制剂。实验方法采用BTK-Ba/F3、BTK(C481S)-Ba/F3细胞和人恶性b细胞JeKo-1、Ramos和NALM-6评价BTK抑制剂的细胞效价。通过细胞活力、菌落形成和btk介导的信号传导来检测体外药理功效和化合物选择性。采用Nu/ Nu BALB/c小鼠BTK-Ba/F3、Ramos和BTK(C481S)-Ba/F3细胞异种肿瘤移植模型，评价XMU-MP-3的体内疗效。XMU-MP-3是一组低分子量化合物之一，是有效的非共价BTK抑制剂。XMU-MP-3在体外和体内均抑制BTK和获得的突变体BTKC481S。进一步的计算模型、定点诱变分析和构效关系研究表明，XMU-MP-3表现出典型的ii型抑制剂结合模式。结论与意义XMU-MP-3在b细胞淋巴瘤中直接靶向BTK信号通路。这些发现表明XMU-MP-3是一种新的BTK抑制剂，可以作为BTK相关b细胞恶性肿瘤的工具化合物和进一步药物开发的导向，特别是那些获得性ibrutinib耐药C481S突变。
Background and Purpose Bruton's tyrosine kinase (BTK) plays a key role in B-cell receptor signalling by regulating cell proliferation and survival in various B-cell malignancies. Covalent low-MW BTK kinase inhibitors have shown impressive clinical efficacy in B-cell malignancies. However, the mutant Btk(C481S) poses a major challenge in the management of B-cell malignancies by disrupting the formation of the covalent bond between BTK and irreversible inhibitors, such as ibrutinib. The present studies were designed to develop novel BTK inhibitors targeting ibrutinib-resistant Btk(C481S) mutation. Experimental Approach BTK-Ba/F3, BTK(C481S)-Ba/F3 cells, and human malignant B-cells JeKo-1, Ramos, and NALM-6 were used to evaluate cellular potency of BTK inhibitors. The in vitro pharmacological efficacy and compound selectivity were assayed via cell viability, colony formation, and BTK-mediated signalling. A tumour xenograft model with BTK-Ba/F3, Ramos and BTK(C481S)-Ba/F3 cells in Nu/nu BALB/c mice was used to assess in vivo efficacy of XMU-MP-3. Key Results XMU-MP-3 is one of a group of low MW compounds that are potent non-covalent BTK inhibitors. XMU-MP-3 inhibited both BTK and the acquired mutant BTKC481S, in vitro and in vivo. Further computational modelling, site-directed mutagenesis analysis, and structure-activity relationships studies indicated that XMU-MP-3 displayed a typical Type-II inhibitor binding mode. Conclusion and Implications XMU-MP-3 directly targets the BTK signalling pathway in B-cell lymphoma. These findings establish XMU-MP-3 as a novel inhibitor of BTK, which could serve as both a tool compound and a lead for further drug development in BTK relevant B-cell malignancies, especially those with the acquired ibrutinib-resistant C481S mutation.