NG25 is a potent and cell-permeable inhibitor of transforming growth factor-β-activated kinase 1 (TAK1) and mitogen-activated protein kinase kinase kinase kinase 2 (MAP4K2), with IC50 values of 149nM and 21.7nM, respectively[1]. NG25 is commonly used in studies related to oncology (e.g., KRAS-mutant colorectal cancer), immune evasion, and tumor microenvironment remodeling[2,3,4].
In vitro, treatment with NG25 (1, 2, 5, 10, and 20μM) for 72h in five breast cancer cell lines (T-47D, MCF7, HCC1954, MDA-MB-231, and BT-549) dose-dependently reduced cell viability in all tested cell lines[4]. Pre-incubation of human aortic vascular smooth muscle cells (HA-VSMCs) with NG25 (0.1, 0.3, 1μM) for 30min, followed by treatment with lipopolysaccharide (LPS, 100ng/mL) for 2h, dose-dependently inhibited LPS-induced phosphorylation of the Smad2 linker region at the Thr220 and Ser245/250/255 sites, with complete inhibition observed at a concentration of 1μM[5].
In vivo, intraperitoneal administration of NG25 (6mg/kg/day) for 2 weeks in mdx mice significantly reduced TAK1 phosphorylation levels and decreased inflammatory cell infiltration in the tibialis anterior and diaphragm muscles[6]. Intravenous administration of NG25 (4mg/kg/day) for 15 days in nude mice bearing Hop62-shCLU (CLU-knockdown Hop62 cell) xenograft tumors slowed tumor growth[7].
References:
[1] Basnet R, Khadka S, Basnet B B, et al. Xanthine oxidase and transforming growth factor beta-activated kinase 1: potential targets for gout intervention[J]. Current Rheumatology Reviews, 2021, 17(2): 153-161.
[2] Xiang Q, Mao Z, Ma Q, et al. NG25 enhances anti-tumor immunity in KRAS-mutant colorectal cancer[J]. OncoTargets and Therapy, 2026: 1-12.
[3] Ma Q, Gu L, Liao S, et al. NG25, a novel inhibitor of TAK1, suppresses KRAS-mutant colorectal cancer growth in vitro and in vivo[J]. Apoptosis, 2019, 24(1): 83-94.
[4] Wang Z, Zhang H, Shi M, et al. TAK1 inhibitor NG25 enhances doxorubicin-mediated apoptosis in breast cancer cells[J]. Scientific Reports, 2016, 6(1): 32737.
[5] Afroz R, Zhou Y, Little P J, et al. Toll-like receptor 4 stimulates gene expression via Smad2 linker region phosphorylation in vascular smooth muscle cells[J]. ACS Pharmacology & Translational Science, 2020, 3(3): 524-534.
[6] Xu D, Li S, Wang L, et al. TAK1 inhibition improves myoblast differentiation and alleviates fibrosis in a mouse model of Duchenne muscular dystrophy[J]. Journal of Cachexia, Sarcopenia and Muscle, 2021, 12(1): 192-208.
[7] Chen Z, Fan Z, Dou X, et al. Inactivation of tumor suppressor gene Clusterin leads to hyperactivation of TAK1-NF-κB signaling axis in lung cancer cells and denotes a therapeutic opportunity[J]. Theranostics, 2020, 10(25): 11520.