通过结合虚拟筛选和NMR光谱方法鉴定了一种新的FGF2 / FGFR信号通路的细胞外抑制剂。
Identification of a novel extracellular inhibitor of FGF2/FGFR signaling axis by combined virtual screening and NMR spectroscopy approach.
发表日期：2023 Apr 07
作者：
Katiuscia Pagano, Roberta Listro, Pasquale Linciano, Daniela Rossi, Elisa Longhi, Giulia Taraboletti, Henriette Molinari, Simona Collina, Laura Ragona
来源：
BIOORGANIC CHEMISTRY
摘要：
非正常激活成纤维细胞生长因子2（FGF2）/成纤维细胞生长因子受体（FGFR）信号通路会导致严重病理学问题，包括癌症发展和血管生成驱动的病理学问题。通过外部变构配体小分子抑制剂干预FGF2/FGFR轴是开发癌症治疗的FGFR抑制剂的有希望的策略，因为这可以提高它们的安全性和有效性。我们之前研究了新的细胞外抑制剂（如迷迭香酸（RA）），它们与FGFR-D2结构域结合并且直接与FGF2竞争相同的结合位点，使功能性FGF2/FGFR相互作用被中断。为了选择先前鉴定的FGF2/FGFR RA结合位点的配体，我们在一套非商业小分子和代谢物的内部库上使用NMR数据驱动虚拟筛选。我们发现了一种新的药物类衍生物，一种叫做RBA4的间苯二酚衍生物。NMR相互作用研究表明，RBA4与FGF2/FGFR复合物结合，与对接预测相符。残基水平的NMR扰动分析强调RBA4与RA的作用机制类似，都能靶向FGF2/FGFR-D2复合物，引起两个蛋白质的扰动，并触发复合物解离。生物学实验证明，RBA4能够抑制FGF2的增殖活性，与先前报道的天然产物RA相当。确定RBA4化学基团参与直接相互作用是进一步优化药物类细胞外抑制剂的起点，以提高其活性。版权所有© 2023 Elsevier Inc.
The aberrant activation of the fibroblast growth factor 2 (FGF2)/fibroblast growth factor receptor (FGFR) signalling pathway drives severe pathologies, including cancer development and angiogenesis-driven pathologies. The perturbation of the FGF2/FGFR axis via extracellular allosteric small inhibitors is a promising strategy for developing FGFR inhibitors with improved safety and efficacy for cancer treatment. We have previously investigated the role of new extracellular inhibitors, such as rosmarinic acid (RA), which bind the FGFR-D2 domain and directly compete with FGF2 for the same binding site, enabling the disruption of the functional FGF2/FGFR interaction. To select ligands for the previously identified FGF2/FGFR RA binding site, NMR data-driven virtual screening has been performed on an in-house library of non-commercial small molecules and metabolites. A novel drug-like compound, a resorcinol derivative named RBA4 has been identified. NMR interaction studies demonstrate that RBA4 binds the FGF2/FGFR complex, in agreement with docking prediction. Residue-level NMR perturbations analysis highlights that the mode of action of RBA4 is similar to RA in terms of its ability to target the FGF2/FGFR-D2 complex, inducing perturbations on both proteins and triggering complex dissociation. Biological assays proved that RBA4 inhibited FGF2 proliferative activity at a level comparable to the previously reported natural product, RA. Identification of RBA4 chemical groups involved in direct interactions represents a starting point for further optimization of drug-like extracellular inhibitors with improved activity.Copyright © 2023 Elsevier Inc. All rights reserved.