Wang Yanqing, Yin Xiaoying, Chen Lingyan, Yin Zhixiang, Zuo Zhicheng
College of Chemistry and Chemical Engineering, Shanghai University of Engineering Science, Shanghai 201620, PR China.
College of Chemistry and Chemical Engineering, Shanghai University of Engineering Science, Shanghai 201620, PR China; Shanghai Frontiers Science Research Center for Druggability of Cardiovascular noncoding RNA, Institute for Frontier Medical Technology, Shanghai University of Engineering Science, Shanghai 201620, PR China; Shanghai Engineering Research Center for Pharmaceutical Intelligent Equipment, Shanghai University of Engineering Science, Shanghai 201620, China.
Bioorg Chem. 2022 Oct;127:105868. doi: 10.1016/j.bioorg.2022.105868. Epub 2022 Jul 2.
Aberrant signaling of EGFR (ErbB) family members, in particular epidermal growth factor receptor (EGFR) and human epidermal growth factor 2 (HER2), is associated with the occurrence and development of many types of human malignancies (e.g., breast, lung, and gastric cancers), and dual targeting of EGFR/HER2 by small-molecular inhibitors has proven to be an effective therapeutic approach for treating these cancers. Herein we extracted and isolated from the medicinal plant Sophora alopecuroides L. a new natural product, dubbed Cytisine N-methylene-(4',7-dihydroxy-3'-methoxy)-isoflavone (CNI1) that features a unique molecular framework. Our biochemical kinase assay suggested that one of its derivative CNI3 exhibited the best, micromolar (μM) inhibition activities against the EGFR (IC of 1.1 μM; K of 0.6 μM) and HER2 (IC of 3.5 μM; K of 1.8 μM) kinases. By contrast, another derivative CNI4 was most potent in inhibiting the EGFR-overexpressing A431 cancer cell line (IC of 45.5 μM) and the HER2-overexpressing BT-474 cancer cell line (IC of 32.9 μM), while the respective cellular activities of Lapatinib (a marketed drug) were 24.9 and 20.3 μM under the same assay condition. Moreover, both CNI3 and CNI4 showed desirable anti-metastatic efficacy in another two breast cancer models (viz., MDA-MB-231 and 4T1). In addition, we explored the inhibitory mechanisms of the CNIs against EGFR and HER2 by molecular dynamics simulation and revealed a novel mode of action that engages the cytisine and chromone moieties in CNIs. By combining structure- and ligand-based analysis, we further rationally engineered a new CNI compound that exhibits considerably improved cytotoxicity against both types of A431 and BT-474 cancer cells. Our study demonstrates the CNI compounds as a new class of EGFR/HER2 dual inhibitors and paves a way for their further development.
表皮生长因子受体（ErbB）家族成员，特别是表皮生长因子受体（EGFR）和人表皮生长因子2（HER2）的异常信号传导，与多种人类恶性肿瘤（如乳腺癌、肺癌和胃癌）的发生和发展相关，并且小分子抑制剂对EGFR/HER2的双重靶向作用已被证明是治疗这些癌症的有效治疗方法。在此，我们从药用植物苦豆子中提取并分离出一种新的天然产物，命名为金雀花碱N-亚甲基-（4'，7-二羟基-3'-甲氧基）-异黄酮（CNI1），其具有独特的分子框架。我们的生化激酶测定表明，其衍生物之一CNI3对EGFR（IC为1.1 μM；K为0.6 μM）和HER2（IC为3.5 μM；K为1.8 μM）激酶表现出最佳的微摩尔（μM）抑制活性。相比之下，另一种衍生物CNI4在抑制EGFR过表达的A431癌细胞系（IC为45.5 μM）和HER过表达的BT-474癌细胞系（IC为32.9 μM）方面最为有效，而在相同测定条件下，拉帕替尼（一种上市药物）的相应细胞活性分别为24.9和20.3 μM。此外，CNI3和CNI4在另外两种乳腺癌模型（即MDA-MB-231和4T1）中均显示出理想的抗转移疗效。此外，我们通过分子动力学模拟探索了CNIs对EGFR和HER2的抑制机制，并揭示了一种涉及CNIs中金雀花碱和色酮部分的新作用模式。通过结合基于结构和配体的分析，我们进一步合理设计了一种新的CNI化合物，该化合物对两种类型的A431和BT-474癌细胞均表现出显著提高的细胞毒性。我们的研究证明了CNI化合物是一类新型的EGFR/HER2双重抑制剂，并为其进一步开发铺平了道路。
