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Updated: May 28, 2025
Antibody-Free Assay for RNA Methyltransferase Activity Analysis
Published on: July 9, 2019
Discovery of a Novel Selective and Cell-Active N6-Methyladenosine RNA Demethylase ALKBH5 Inhibitor
Xianyuan Yang1, Kaitao Huang1, Xu-Nian Wu1
1Guangdong Provincial Key Laboratory of Chiral Molecule and Drug Discovery, State Key Laboratory of Anti-Infective Drug Discovery and Development, School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou 510006, China.
Abstract:
N6-methyladenosine (m6A), the most abundant methylation on mRNA, plays pivotal roles in regulating mRNA biological functions, which affect cell functions. ALKBH5, an m6A demethylase, was found to be an oncogene in several cancer types, including triple-negative breast cancer (TNBC). Here, we report a novel and selective ALKBH5 covalent inhibitor, W23-1006, through virtual screening and structure optimization. It covalently bonds to the ALKBH5 C200 residue with an IC50 value of 3.848 μM, representing roughly 30- and 8-fold stronger inhibitory activity than that against FTO and ALKBH3, respectively. Cellular experiments demonstrated that W23-1006 could efficiently enhance the m6A level on fibronectin 1 (FN1) mRNA, leading to strong suppression of TNBC cell proliferation and migration in vitro as well as tumor growth and metastasis in vivo. Collectively, our study developed a novel, selective, and cell-active ALKBH5 covalent inhibitor, W23-1006, which could be a potential therapeutic option for cancer, such as TNBC treatment.
Insights
Researchers developed W23-1006, a selective inhibitor targeting ALKBH5, an oncogene in triple-negative breast cancer (TNBC). This compound suppresses TNBC cell proliferation and tumor growth by increasing mRNA methylation.
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- N 6-methyladenosine (m 6 A) is the most prevalent mRNA modification, regulating gene expression and cellular functions.
- ALKBH5, an m 6 A demethylase, functions as an oncogene in various cancers, including triple-negative breast cancer (TNBC).
Purpose of the Study:
- To develop a novel, selective, and potent ALKBH5 covalent inhibitor.
- To evaluate the therapeutic potential of the inhibitor in TNBC models.
Main Methods:
- Virtual screening and structure-based optimization were employed to identify and refine the inhibitor.
- In vitro assays determined the inhibitory activity (IC 50) against ALKBH5, FTO, and ALKBH3.
- Cellular and in vivo experiments assessed the compound's efficacy in suppressing TNBC progression.
Main Results:
- A selective ALKBH5 covalent inhibitor, W23-1006, was identified with an IC 50 of 3.848 μM.
- W23-1006 demonstrated significantly higher potency against ALKBH5 compared to FTO and ALKBH3.
- The inhibitor effectively increased m 6 A levels on fibronectin 1 (FN1) mRNA, inhibiting TNBC cell proliferation, migration, tumor growth, and metastasis.
Conclusions:
- W23-1006 is a novel, selective, and cell-active covalent inhibitor of ALKBH5.
- This inhibitor shows promise as a potential therapeutic agent for TNBC and potentially other cancers driven by ALKBH5.
