Discovery of METTL3 Small Molecule Inhibitors by Virtual Screening of Natural Products.
Discovery of METTL3 Small Molecule Inhibitors by Virtual Screening of Natural Products.
复制标题
通过天然产物虚拟筛选发现 METTL3 小分子抑制剂
DOI：
10.3389/fphar.2022.878135
复制
发表时间：
2022
影响因子：
5.6
通讯作者：
Tian, Xin
中科院分区：
文献类型：
--
作者：
Du, Yue;Yuan, Yongliang;Xu, Le;Zhao, Fang;Wang, Wenbin;Xu, Yiping;Tian, Xin
N6-甲基腺苷（m6 A）是哺乳动物细胞中最常见的mRNA修饰，其主要由甲基转移酶样3和甲基转移酶样14（肌L3-肌L14）的甲基转移酶复合物催化。许多证据表明，胃L3在癌症和病毒感染等多种疾病中起着重要作用。本研究选取了1,042种市售天然产物建立筛选文库，并进行了基于对接的高通量筛选，以发现潜在的胃L3抑制剂。然后通过体外甲基转移酶抑制测定进一步验证所选化合物，其中通过LC-MS/MS测定m6 A含量。进行m6 A甲基化抑制的细胞测定以测定所选化合物的胃L3抑制活性。CCK-8测定用于评价所选化合物对肿瘤细胞活力的影响。此外，结合模式分析，分子动力学（MD）模拟，结合自由能分析，研究抑制剂结合的过程和特点。最后，槲皮素被鉴定为胃L3抑制剂，IC 50值为2.73 μM。m6 A甲基化抑制的细胞测定显示，槲皮素以剂量依赖性方式降低MIA PaCa-2胰腺癌细胞中m6 A的水平。CCK-8法显示槲皮素能有效抑制MIA PaCa-2和Huh 7肿瘤细胞的增殖，IC_（50）分别为73.51 ± 11.22 μM和99.97 ± 7.03 μM。分子对接研究表明，槲皮素填补了口袋的腺苷部分的SAM，但不是口袋的SAM蛋氨酸在胃L3蛋白，和氢键，疏水相互作用，和π堆积形成。均方根偏差（RMSD）、均方根波动（RMSF）和结合自由能表明槲皮素可以有效地结合到胃L3蛋白的口袋中，并形成稳定的蛋白-配体复合物。本研究首次从天然产物中鉴定出了胃L3抑制剂，为后续研究提供了基础，有利于开发以胃L3为靶点的疾病治疗药物。
N6-Methyladenosine (m6A) is the most prevalent mRNA modification in mammalian cells that is mainly catalyzed by the methyltransferase complex of methyltransferase-like 3 and methyltransferase-like 14 (METTL3-METTL14). Many lines of evidence suggest that METTL3 plays important roles in several diseases such as cancers and viral infection. In the present study, 1,042 natural products from commercially available sources were chosen to establish a screening library, and docking-based high-throughput screening was performed to discover potential METTL3 inhibitors. The selected compounds were then further validated by an in vitro methyltransferase inhibition assay in which m6A content was determined by LC-MS/MS. A cellular assay of the inhibition of m6A methylation was performed to determine the METTL3 inhibitory activity of the selected compound. CCK-8 assay was applied to evaluate the effects of the selected compound on tumor cell viability. Additionally, binding mode analysis, molecular dynamics (MD) simulation, and binding free energy analysis were performed to study the process and characteristics of inhibitor binding. Finally, quercetin was identified as a METTL3 inhibitor with an IC50 value of 2.73 μM. The cellular assay of m6A methylation inhibition showed that quercetin decreased m6A level in a dose-dependent manner in MIA PaCa-2 pancreatic cancer cells. CCK-8 assay showed quercetin efficiently inhibited the proliferation of MIA PaCa-2 and Huh7 tumor cells, with IC50 values 73.51 ± 11.22 μM and 99.97 ± 7.03 μM, respectively. Molecular docking studies revealed that quercetin filled the pocket of the adenosine moiety of SAM but not the pocket of the SAM methionine in the METTL3 protein, and hydrogen bonds, hydrophobic interactions, and pi-stacking were formed. The values of the root mean square deviation (RMSD), the root mean square fluctuations (RMSF), and binding free energy suggested that quercetin can efficiently bind to the pocket of the METTL3 protein and form a stable protein-ligand complex. The present study is the first to identify METTL3 inhibitors from natural products, thus providing a basis for subsequent research and facilitating the development of METTL3-targeting drugs for diseases.
登录
查看更多内容
n（6） - 甲基腺苷依赖性RNA结构开关调节RNA蛋白质相互作用。
影响因子：
64.8
作者：
Liu, Nian;Dai, Qing;Zheng, Guanqun;He, Chuan;Parisien, Marc;Pan, Tao
通讯作者：
Pan, Tao
影响因子：
3.4
作者：
Moroz-Omori EV;Huang D;Kumar Bedi R;Cheriyamkunnel SJ;Bochenkova E;Dolbois A;Rzeczkowski MD;Li Y;Wiedmer L;Caflisch A
通讯作者：
Caflisch A
m(6)A mRNA甲基化调控CTNNB1促进肝母细胞瘤增殖
影响因子：
37.3
作者：
Liu, Li;Wang, Jing;Pan, Qiuhui
通讯作者：
Pan, Qiuhui
影响因子：
7.3
作者：
Dolbois, Aymeric;Bedi, Rajiv K.;Caflisch, Amedeo
通讯作者：
Caflisch, Amedeo
DOI：
10.1080/01635581.2016.1145245
发表时间：
2016-02-17
影响因子：
2.9
作者：
Brito, Ana Filipa;Ribeiro, Marina;Botelho, Maria Filomena
通讯作者：
Botelho, Maria Filomena