BML-284 (Wnt agonist 1) is a potent agonist of Wnt signaling, with an EC50 value of 0.7μM for inducing β-catenin- and TCF-dependent transcriptional activity[1]. BML-284 binds to the colchicine-site and forms a low barrier hydrogen bond with E198 of β-tubulin, to mediate αβ-tubulin degradation through the ubiquitin-proteasome pathway[2]. BML-284 has been widely used as a model compound to synthesize a series of photosensitive azo derivatives that undergo reversible cis-trans isomerization upon visible light irradiation, with the cis-isomer capable of activating Wnt signaling [3].
In vitro, BML-284 treatment for 72 hours significantly inhibited the proliferation of HeLa, HCT116, MCF-7 and K562 cells with IC50 values of 0.09µM, 0.52µM, 0.69µM and 0.10µM, respectively[4]. Treatment with 1µM BML-284 for 24 hours promoted invasion and inhibited apoptosis of A549 cells transfected with pcDNA-AOC4P[5]. Pre-incubation of human aortic vascular smooth muscle cells (HA-VSMCs) with 10µM BML-284 for 24 hours activated the Wnt signaling pathway, increased the protein expression of Wnt3a and β-catenin, and reversed the protective effects of si-SOX2 on calcification, osteogenic/contractile protein expression, and autophagy activity[6].
In vivo, BML-284 treatment (5μl; 0.5μg/eye) via injection into the conjunctiva adjacent to the limbus for 48 hours promoted corneal epithelial healing and corneal nerve repair in diabetic mice[7]. BML-284 treatment via daily intravenous injection at a dose of 500µl (2mg/ml) for 14 days antagonized the protective effect of Triptolide (TP) against myocardial fibrosis (MF) by activating the Wnt/β-catenin pathway, and aggravated cardiac dysfunction in rats[8].
References:
[1] Liu J, Wu X, Mitchell B, et al. A small‐molecule agonist of the Wnt signaling pathway[J]. Angewandte Chemie International Edition, 2005, 44(13): 1987-1990.
[2] Yang J, Li Y, Qiu Q, et al. Small molecules promote selective denaturation and degradation of tubulin heterodimers through a low-barrier hydrogen bond[J]. Journal of Medicinal Chemistry, 2022, 65(13): 9159-9173.
[3] Ahmad S, Hashim P K, Imajo M, et al. Photoswitchable agonists for visible-light activation of the Wnt signaling pathway[J]. Organic & Biomolecular Chemistry, 2025, 23(17): 4240-4245.
[4] Zhang C, Yan W, Liu Y, et al. Structure-based design and synthesis of BML284 derivatives: A novel class of colchicine-site noncovalent tubulin degradation agents[J]. European Journal of Medicinal Chemistry, 2024, 268: 116265.
[5] Li F, Rong T, Cao G, et al. AOC4P suppresses viability and invasion and induces apoptosis in NSCLC cells by inhibiting the Wnt/β-catenin pathway[J]. Chemico-Biological Interactions, 2020, 325: 109110.
[6] Xiang H, Hong J, Zhang Y, et al. Inhibition of SOX2 mitigates vascular calcification mediated by autophagy through Wnt signaling pathway[J]. Egyptian Journal of Medical Human Genetics, 2025, 26(1): 189.
[7] Yang S, Zhang Y, Zhang Z, et al. Insulin promotes corneal nerve repair and wound healing in type 1 diabetic mice by enhancing Wnt/β-catenin signaling[J]. The American Journal of Pathology, 2020, 190(11): 2237-2250.
[8] Zhang Y, Lu F. Molecular mechanism of triptolide in myocardial fibrosis through the Wnt/β-catenin signaling pathway[J]. Scandinavian Cardiovascular Journal, 2024, 58(1): 2295785.
BML-284 (Wnt agonist 1)是一种Wnt信号通路的强效激动剂，诱导β-catenin和TCF依赖性转录活性的EC50值为0.7μM[1]。BML-284结合在秋水仙碱位点，并与β-微管蛋白的E198形成低能氢键，从而通过泛素-蛋白酶体途径介导αβ-微管蛋白降解[2]。BML-284已广泛用作模型化合物，用于合成一系列在可见光照射下可发生可逆顺反异构化的光敏偶氮衍生物，其中顺式异构体能够激活Wnt信号通路[3]。
在体外，BML-284处理72小时显著抑制了HeLa、HCT116、MCF-7和K562细胞的增殖，IC50值分别为0.09µM、0.52µM、0.69µM和0.10µM[4]。用1µM的BML-284处理24小时促进了转染pcDNA-AOC4P的A549细胞的侵袭并抑制了细胞凋亡[5]。用10µM的BML-284预孵育人主动脉血管平滑肌细胞（HA-VSMCs）24小时激活了Wnt信号通路，增加了Wnt3a和β-catenin的蛋白表达，并逆转了si-SOX2对钙化、成骨/收缩蛋白表达和自噬活性的保护作用[6]。
在体内，向角膜缘附近的结膜注射BML-284（5μl；0.5μg/眼）处理48小时促进了糖尿病小鼠的角膜上皮愈合和角膜神经修复[7]。每日静脉注射500µl（2mg/ml）剂量的BML-284连续14天，通过激活Wnt/β-catenin通路拮抗了Triptolide（TP）对心肌纤维化（MF）的保护作用，并加重了大鼠的心脏功能障碍[8]。