四川大学杨胜勇团队报道了DNA 6mA去甲基化酶ALKBH1有效细胞活性抑制剂的发现。相关研究成果于2024年3月4日发表在《美国化学会杂志》。

DNA的N6-甲基腺嘌呤（6mA）已成为真核生物中一种新的表观遗传学标记，并已鉴定出几种6mA效应蛋白。然而，迄今为止，用小分子选择性抑制这些效应蛋白的生物功能的努力是不成功的。

该文中，研究人员报道了AlkB同源物1（ALKBH1）的第一个有效和选择性小分子抑制剂（13h），这是唯一经过验证的6mA去甲基酶。13h在荧光偏振（FP）和酶活性测定中的IC50分别为0.026±0.013μM和1.39±0.13μM，在等温滴定量热法（ITC）测定中的KD为0.112±0.017μM。13h-ALKBH₁配合物的共晶结构很好地解释了13h的效力。此外，13h对ALKBH₁表现出优异的选择性。在细胞中，化合物13h及其衍生物16能够与ALKBH₁结合并调节6mA水平。

总之，该研究确定了第一种有效的、同种型选择性的、具有细胞活性的ALKBH1抑制剂，为探索ALKBH1和DNA 6mA的生物学功能提供了一种工具化合物。

附：原文原文

Title: Discovery of a Potent and Cell-Active Inhibitor of DNA 6mA Demethylase ALKBH1

Author: Liang Xiong, Feng Li, Yinping Guo, Jian Zhang, Ke Xu, Zijie Xiong, Aiping Tong, Linli Li, Shengyong Yang

Issue&Volume: March 4, 2024

Abstract: N6-Methyladenine (6mA) of DNA has emerged as a novel epigenetic mark in eukaryotes, and several 6mA effector proteins have been identified. However, efforts to selectively inhibit the biological functions of these effector proteins with small molecules are unsuccessful to date. Here we report the first potent and selective small molecule inhibitor (13h) of AlkB homologue 1 (ALKBH1), the only validated 6mA demethylase. 13h showed an IC50 of 0.026 ± 0.013 μM and 1.39 ± 0.13 μM in the fluorescence polarization (FP) and enzyme activity assay, respectively, and a KD of 0.112 ± 0.017 μM in the isothermal titration calorimetry (ITC) assay. The potency of 13h was well explained by the cocrystal structure of the 13h-ALKBH1 complex. Furthermore, 13h displayed excellent selectivity for ALKBH1. In cells, compound 13h and its derivative 16 were able to engage ALKBH1 and modulate the 6mA levels. Collectively, our study identified the first potent, isoform selective, and cell-active ALKBH1 inhibitor, providing a tool compound for exploring the biological functions of ALKBH1 and DNA 6mA.

DOI: 10.1021/jacs.4c00194

Source: https://pubs.acs.org/doi/abs/10.1021/jacs.4c00194