Volpe Serena Della, Listro Roberta, Ambrosio Francesca Alessandra, Garbagnoli Martina, Linciano Pasquale, Rossi Daniela, Costa Giosuè, Alcaro Stefano, Vasile Francesca, Hirsch Anna K H, Collina Simona
Department of Drug Sciences, University of Pavia, Via Taramelli 12, 27100 Pavia, Italy.
Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Centre for Infection Research (HZI), Campus E8.1, 66123 Saarbrücken, Germany.
ACS Med Chem Lett. 2023 Oct 6;14(11):1509-1516. doi: 10.1021/acsmedchemlett.3c00303. eCollection 2023 Nov 9.
The RNA binding protein HuR regulates the post-transcriptional process of different oncogenes and tumor suppressor genes, and its dysregulation is linked with cancer. Thus, modulating the complex HuR-RNA represents a promising anticancer strategy. To search for novel HuR ligands able to interfere with the HuR-RNA complex, the protein-templated dynamic combinatorial chemistry (pt-DCC) method was utilized. The recombinant RRM1+2 protein construct, which contains essential domains for ligand-HuR binding and exhibits enhanced solubility and stability compared to the native protein, was used for pt-DCC. Seven acylhydrazones with over 80% amplification were identified. The binding of the fragments to HuR extracted from DCC was validated using STD-NMR, and molecular modeling studies revealed the ability of the compounds to bind HuR at the mRNA binding pocket. Notably, three compounds effectively interfered with HuR-RNA binding in fluorescence polarization studies, suggesting their potential as foundational compounds for developing anticancer HuR-RNA interfering agents.
RNA结合蛋白HuR调节不同癌基因和肿瘤抑制基因的转录后过程，其失调与癌症相关。因此，调节HuR-RNA复合物是一种很有前景的抗癌策略。为了寻找能够干扰HuR-RNA复合物的新型HuR配体，采用了蛋白质模板动态组合化学（pt-DCC）方法。重组RRM1+2蛋白构建体用于pt-DCC，该构建体包含配体与HuR结合的必需结构域，并且与天然蛋白相比具有更高的溶解度和稳定性。鉴定出七种扩增率超过80%的酰腙。使用STD-NMR验证了从DCC中提取的片段与HuR的结合，分子建模研究揭示了这些化合物在mRNA结合口袋处结合HuR的能力。值得注意的是，在荧光偏振研究中，三种化合物有效地干扰了HuR-RNA结合，表明它们作为开发抗癌HuR-RNA干扰剂的基础化合物的潜力。
