Phenylpyrazoles as Inhibitors of the m<sup>6</sup>A RNA-Binding Protein YTHDF2
Xiaqiu Qiu, Claus Kemker, Georg L. Goebel, Philipp Lampe, Nadav Wallis, Damian Schiller, Katrin Bigler, Mao Jiang, Sonja Sievers, G Yeo, Peng Wu
Abstract
High Resolution Image Download MS PowerPoint Slide The N 6-methyladenosine (m 6 A) modification, which is the most common RNA modification in eukaryotes, is regulated by the “writer” methyltransferases, the “reader” m 6 A binding proteins, and the “eraser” demethylases. m 6 A plays a multifunctional role in physiological and pathological processes, regulating all aspects of RNA metabolism and function, including RNA splicing, translation, transportation, and degradation. Accumulating evidence suggests that the YT521-B homology domain family 2 (YTHDF2), one of the m 6 A “readers,” is associated with various biological processes in cancers and noncancerous disorders, impacting migration, invasion, metastasis, proliferation, apoptosis, and cell cycle. Here, we describe our work in the identification of a series of functionalized pyrazoles, such as CK-75, as new YTHDF2 inhibitors, which potentially bind to a small hydrophobic pocket on the YTH domain. Cellular evaluations revealed that the small-molecule YTHDF2 inhibitors induced cell cycle arrest, induced apoptosis, and significantly inhibited the cell viability of cancer cells. Furthermore, we evaluated the transcriptome-wide change in the global RNA-binding protein and RNA-binding patterns of CK-75 via an enhanced cross-linking and immunoprecipitation assay. Our work demonstrated the feasibility of targeting the YTH domain of YTHDF2 with small molecules. The phenylpyrazoles studied in this work provided a lead structure for the further development of small molecules targeting YTHDF2 for both biological and therapeutic applications.