Recently, the research team led by Huang Zhangjian and Wu Jianbing from our university published their latest research findings in the prestigious journal Journal of the American Chemical Society, titled “Warhead Strategy for Targeted Protein S-Nitrosation.” The paper's co-first authors are Zhang Chen, a 2022 doctoral student at our university, Ye Hui, a postdoctoral researcher, and Ji Duorui, a 2022 doctoral student. The corresponding authors are Huang Zhangjian, a researcher at our university, and Wu Jianbing, an associate researcher. China Pharmaceutical University is the first corresponding institution for the paper.
Research on protein post-translational modifications plays a crucial role in drug development. Nitric oxide (NO) is a gaseous free radical signaling molecule, and NO-mediated protein S-nitrosation (SNO) is an important post-translational modification, formed by the reaction between NO and cysteine sulfhydryl groups to generate SNO groups. Protein nitrosylation can influence protein function, protein-protein interactions, and other post-translational modifications, mediating diverse biological effects. Research into the relationship between the nitrosylation of specific protein key sites and disease onset and progression is one of the frontiers in the field of dynamic modifications of biomacromolecules. However, how to use medicinal chemistry methods to precisely intervene in the nitrosylation of specific protein key sites to produce disease-treating activity remains an unexplored area. The main challenge lies in the fact that NO has the property of gaseous free diffusion, making it difficult for existing therapies to restrict its action to specific proteins.
Addressing these key scientific issues, the team employed a covalent inhibitor strategy to design and synthesize a series of NO-releasing warheads, which were then incorporated into the small-molecule ligand cores targeting kinases BTK, FGFR4, and HER2. This enables the specific sulfhydryl groups of the target proteins to form covalent bonds with the warheads while releasing NO, which distributes within an extremely small range and selectively nitrosylates the target proteins themselves. The results showed that the selective nitrosation modulator TSNO1-8 exhibits superior in vitro stability, good pharmacokinetic properties, excellent kinase spectrum selectivity, and superior in vitro and in vivo anti-lymphoma activity. Further nitrosylation omics studies indicate that the selective nitrosylation modulator TSNO1 can selectively nitrosylate Cys527 of BTK both in vivo and in vitro. The nitrosylation modification at this site disrupts the conformation of the BTK kinase domain, causing Tyr551 to be hydrogen-bonded, thereby further impairing BTK's kinase signal transduction. In summary, this study not only provides a new method for selective nitrosylation regulation but also offers new insights into the development of selective post-translational modification drugs.
This research was supported by the National Natural Science Foundation of China, the “Double First-Class” Construction Fund of China Pharmaceutical University, and the Xingyao Leading Scholar Fund.
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