Manabe Yoshiyuki, Takebe Tomoyuki, Kasahara Satomi, Hizume Koki, Kabayama Kazuya, Kamada Yoshihiro, Asakura Akiko, Shinzaki Shinichiro, Takamatsu Shinji, Miyoshi Eiji, García-García Ana, Vakhrushev Sergey Y, Hurtado-Guerrero Ramón, Fukase Koichi
Department of Chemistry, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka, 560-0043, Japan.
Forefront Research Center, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka, 560-0043, Japan.
Angew Chem Int Ed Engl. 2024 Dec 20;63(52):e202414682. doi: 10.1002/anie.202414682. Epub 2024 Nov 6.
Core fucosylation is catalyzed by α-1,6-fucosyltransferase (FUT8), which fucosylates the innermost GlcNAc of N-glycans. Given the association of FUT8 with various diseases, including cancer, selective FUT8 inhibitors applicable to in vivo or cell-based systems are highly sought-after. Herein, we report the discovery of a compound that selectively inhibits FUT8 in cell-based assays. High-throughput screening revealed a FUT8-inhibiting pharmacophore, and further structural optimization yielded an inhibitor with a K value of 49 nM. Notably, this binding occurs only in the presence of GDP (a product of the enzymatic reaction catalyzed by FUT8). Mechanistic studies suggested that this inhibitor generates a highly reactive naphthoquinone methide derivative at the binding site in FUT8, which subsequently reacts with FUT8. Furthermore, prodrug derivatization of this inhibitor improved its stability, enabling suppression of core fucose expression and subsequent EGFR and T-cell signaling in cell-based assays, paving the way for the development of drugs targeting core fucosylation.
核心岩藻糖基化由α-1,6-岩藻糖基转移酶（FUT8）催化，该酶可将岩藻糖基连接到N-聚糖最内层的N-乙酰葡糖胺上。鉴于FUT8与包括癌症在内的多种疾病相关，因此人们迫切需要适用于体内或基于细胞系统的选择性FUT8抑制剂。在此，我们报告了一种在基于细胞的实验中能选择性抑制FUT8的化合物的发现。高通量筛选揭示了一种FUT8抑制药效团，进一步的结构优化得到了一种K值为49 nM的抑制剂。值得注意的是，这种结合仅在GDP（FUT8催化的酶促反应的产物）存在时发生。机理研究表明，该抑制剂在FUT8的结合位点产生一种高反应性的萘醌甲基化物衍生物，随后与FUT8发生反应。此外，该抑制剂的前药衍生化提高了其稳定性，使其能够在基于细胞的实验中抑制核心岩藻糖表达以及随后的表皮生长因子受体（EGFR）和T细胞信号传导，为开发针对核心岩藻糖基化的药物铺平了道路。
