Jiang Shilong, Liu Keyi, Jiang Ting, Li Hui, Wei Xiao, Wan Xiaoya, Zhong Changxin, Gong Rong, Chen Zonglin, Zou Chan, Zhang Qing, Cheng Yan, Cao Dongsheng
Department of Pharmacy, Xiangya Hospital, Central South University, Changsha, China.
The Hunan Institute of Pharmacy Practice and Clinical Research, Changsha, Hunan, China.
Nat Commun. 2025 Aug 22;16(1):7854. doi: 10.1038/s41467-025-62288-7.
Target identification in natural products plays a critical role in the development of innovative drugs. Bufalin, a compound derived from traditional medicines, has shown promising anti-cancer activity; however, its precise molecular mechanism of action remains unclear. Here, we employ artificial intelligence, molecular docking, and molecular dynamics simulations to elucidate the molecular mechanism of Bufalin. Using an integrated multi-predictive strategy, we identify CYP17A1, ESR1, mTOR, AR, and PRKCD as the potential targets of Bufalin. Subsequent validation via surface plasmon resonance, biotin pulldown, and thermal shift assays confirms Bufalin's direct binding to ESR1, which encodes estrogen receptor alpha (ERα). Molecular docking analyses pinpoint Bufalin's selective interaction with Arg394 on ERα. Molecular dynamic simulations further show that Bufalin acts as a molecular glue, enhancing the interaction between ERα and the E3 ligase STUB1, thereby promoting proteasomal degradation of ERα. Given the therapeutic potential of ERα degradation in overcoming endocrine resistance, we investigate the inhibitory effect of Bufalin on endocrine-resistant models and prove Bufalin reverses Tamoxifen resistance in vitro, in vivo, and in patient-derived breast cancer organoids from tamoxifen-relapsed cases. Collectively, our findings indicate that Bufalin functions as a molecular glue to degrade ERα, offering a potential therapeutic strategy for reversing Tamoxifen resistance.
天然产物中的靶点识别在创新药物研发中起着关键作用。蟾毒灵是一种从传统药物中提取的化合物，已显示出有前景的抗癌活性；然而，其确切的分子作用机制仍不清楚。在此，我们采用人工智能、分子对接和分子动力学模拟来阐明蟾毒灵的分子机制。通过综合多种预测策略，我们确定细胞色素P450 17A1（CYP17A1）、雌激素受体1（ESR1）、雷帕霉素靶蛋白（mTOR）、雄激素受体（AR）和蛋白激酶Cδ（PRKCD）为蟾毒灵的潜在靶点。随后通过表面等离子体共振、生物素下拉和热位移分析进行验证，证实蟾毒灵与编码雌激素受体α（ERα）的ESR1直接结合。分子对接分析确定了蟾毒灵与ERα上的精氨酸394（Arg394）的选择性相互作用。分子动力学模拟进一步表明，蟾毒灵作为一种分子胶水，增强了ERα与E3连接酶STUB1之间的相互作用，从而促进ERα的蛋白酶体降解。鉴于ERα降解在克服内分泌耐药性方面的治疗潜力，我们研究了蟾毒灵对内分泌耐药模型的抑制作用，并证明蟾毒灵在体外、体内以及来自他莫昔芬复发病例的患者来源的乳腺癌类器官中逆转了他莫昔芬耐药性。总的来说，我们的研究结果表明，蟾毒灵作为一种分子胶水来降解ERα，为逆转他莫昔芬耐药性提供了一种潜在的治疗策略。
