Song Yuqing, Zhang Feng, Guo Jia, Fan Yufan, Zeng Hairong, Sun Mengru, Qian Jun, Qi Shenglan, Chen Zihan, Jin Xudong, Song Yunqing, Tian Tian, Qian Zhi, Sun Yao, Tian Zhenhao, Yu Baoqing, Ge Guangbo
State Key Laboratory of Discovery and Utilization of Functional Components in Traditional Chinese Medicine, Shanghai Frontiers Science Center of TCM Chemical Biology, Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
Department of Orthopedics, the Seventh People's Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200137, China.
Acta Pharm Sin B. 2025 Aug;15(8):4174-4192. doi: 10.1016/j.apsb.2025.06.004. Epub 2025 Jun 9.
Notum, a negative feedback regulator of the Wnt signaling, has emerged as a promising target for treating glucocorticoid-induced osteoporosis (GIOP). This study showcases an efficient strategy for discovering the anti-Notum constituents from herbal medicines (HMs) as novel anti-GIOP agents. Firstly, a rapid-responding near-infrared fluorogenic substrate for Notum was rationally engineered for high-throughput identifying the anti-Notum HMs. The results showed that Bu-Gu-Zhi (BGZ), a known anti-osteoporosis herb, potently inhibited Notum in a competitive-inhibition manner. To uncover the key anti-Notum constituents in BGZ, an efficient strategy was adapted integrating biochemical, phytochemical, computational, and pharmacological assays. Among all identified BGZ constituents, three furanocoumarins were validated as strong Notum inhibitors, while 5-methoxypsoralen (5-MP) showed the most potent anti-Notum activity and favorable safety profiles. Mechanistically, 5-MP acted as a competitive inhibitor of Notum creating strong hydrophobic interactions with Trp128 and Phe268 in the catalytic cavity of Notum. Cellular assays showed that 5-MP remarkably promoted osteoblast differentiation and activated Wnt signaling in dexamethasone (DXMS)-challenged MC3T3-E1 osteoblasts. In dexamethasone-induced osteoporotic mice, 5-MP strongly elevated bone mineral density (BMD) and improved cancellous and cortical bone thickness. Collectively, this study constructs a high-efficient platform for discovering key anti-Notum constituents from HMs, while 5-MP emerges as a promising anti-GIOP agent.
Notum作为Wnt信号通路的负反馈调节因子，已成为治疗糖皮质激素诱导的骨质疏松症（GIOP）的一个有前景的靶点。本研究展示了一种从草药（HMs）中发现抗Notum成分作为新型抗GIOP药物的有效策略。首先，合理设计了一种用于Notum的快速响应近红外荧光底物，用于高通量鉴定抗Notum的草药。结果表明，已知的抗骨质疏松草药补骨脂（BGZ）以竞争性抑制方式有效抑制Notum。为了揭示BGZ中的关键抗Notum成分，采用了一种整合生化、植物化学、计算和药理学分析的有效策略。在所有鉴定出的BGZ成分中，三种呋喃香豆素被证实为强效的Notum抑制剂，而5-甲氧基补骨脂素（5-MP）表现出最有效的抗Notum活性和良好的安全性。从机制上讲，5-MP作为Notum的竞争性抑制剂，与Notum催化腔中的Trp128和Phe268形成强烈的疏水相互作用。细胞实验表明，5-MP显著促进了地塞米松（DXMS）刺激的MC3T3-E1成骨细胞的成骨细胞分化并激活了Wnt信号通路。在去甲肾上腺素诱导的骨质疏松小鼠中，5-MP显著提高了骨密度（BMD），并改善了松质骨和皮质骨厚度。总的来说，本研究构建了一个从草药中发现关键抗Notum成分的高效平台，而5-MP成为一种有前景的抗GIOP药物。
