<br>METTL3–14 m6A-RNA 甲基转移酶嵌合体降解剂的蛋白水解作用,JACS Au

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METTL3–14 m6A-RNA 甲基转移酶嵌合体降解剂的蛋白水解作用
JACS Au ( IF 8.5 ) Pub Date : 2024-02-12 , DOI: 10.1021/jacsau.4c00040
Francesco Errani ₁ , Annalisa Invernizzi ₁ , Marcin Herok ₁ , Elena Bochenkova ₁ , Fiona Stamm ₁ , Ivan Corbeski ₁ , Valeria Romanucci ₂ , Giovanni Di Fabio ₂ , František Zálešák ₁ , Amedeo Caflisch ₁

Affiliation

腺嘌呤 N6 (m ⁶ A) 的甲基化是最常见的 RNA 修饰。在 mRNA 上，它由 METTL3-14 异二聚体复合物催化，该复合物在急性髓系白血病 (AML) 以及其他类型的血癌和实体瘤中发挥着关键作用。在这里，我们公开了第一个针对表观转录组蛋白的蛋白水解靶向嵌合体（PROTAC）。为了设计 PROTAC，我们利用了 METTL3-14 与有效的选择性小分子抑制剂（称为 UZH2）复合物的晶体结构。连接子的优化从UZH2的脱氟前体开始，其合成比UZH2更有效。前九个 PROTAC 分子具有基于 PEG 或烷基的接头，但只有后者显示出细胞渗透性。有了这些信息，我们合成了 26 个基于 UZH2 和不同长度和刚性的烷基连接体的 PROTAC。三元复合物的形成通过基于 FRET 的生化测定和体外泛素化测定进行了验证。通过蛋白质印迹在MOLM-13细胞中测量，具有不同接头类型和长度的PROTAC 14、20、22、24和30显示出50%或更高的METTL3和/或METTL14降解。它们还显示出对其他三种 AML 细胞系和前列腺癌细胞系 PC3 的显着降解。

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Proteolysis Targeting Chimera Degraders of the METTL3–14 m6A-RNA Methyltransferase

Methylation of adenine N6 (m⁶A) is the most frequent RNA modification. On mRNA, it is catalyzed by the METTL3–14 heterodimer complex, which plays a key role in acute myeloid leukemia (AML) and other types of blood cancers and solid tumors. Here, we disclose the first proteolysis targeting chimeras (PROTACs) for an epitranscriptomics protein. For designing the PROTACs, we made use of the crystal structure of the complex of METTL3–14 with a potent and selective small-molecule inhibitor (called UZH2). The optimization of the linker started from a desfluoro precursor of UZH2 whose synthesis is more efficient than that of UZH2. The first nine PROTAC molecules featured PEG- or alkyl-based linkers, but only the latter showed cell penetration. With this information in hand, we synthesized 26 PROTACs based on UZH2 and alkyl linkers of different lengths and rigidity. The formation of the ternary complex was validated by a FRET-based biochemical assay and an in vitro ubiquitination assay. The PROTACs 14, 20, 22, 24, and 30, featuring different linker types and lengths, showed 50% or higher degradation of METTL3 and/or METTL14 measured by Western blot in MOLM-13 cells. They also showed substantial degradation on three other AML cell lines and prostate cancer cell line PC3.

更新日期：2024-02-12

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