ATN-224, the bis-choline salt of tetrathiomolybdate (TM), is a superoxide dismutase inhibitor and specific, high-affinity copper binder that suppresses CuZn superoxide dismutase 1 (SOD1) activity in blood and endothelial cells, leading to antiangiogenic and antitumour effects[1-3]. ATN-224 inhibited Fibroblast growth factor-2 (FGF-2)-induced extracellular signal-regulated kinases 1 and 2 (ERK1/2) phosphorylation in a dose-dependent (0, 0.625, 1.25, 2.5, 5, 10μM) and time-dependent (0, 1, 2, 4, 6, 8, 16, 24, 48h) manner with an IC50 between 1.25 and 2.5μM[4].
In vitro, treatment of WEHI7.2 and its variants, namely, hydrogen-peroxide-resistant 200R cells and Bcl-2-overexpressing Hb12 cells, with 0-10nM ATN-224 for 48h decreased the number of viable cells in a concentration-dependent manner, yielding EC50 values of 3.17±0.27, 5.84±0.34, and 5.25±0.32nM, respectively. Concurrently, ATN-224 markedly elevated caspase-3 activity and propidium iodide (PI) uptake in all three cell lines[5]. Treatment of SUDHL-10 cells with 32.71±2.04nM ATN-224 caused a significant drop in mitochondrial membrane potential (ΔΨm) within 12h and a pronounced reduction in Mcl-1 protein levels by 48h[6].
In vivo, in Matrigel plug model female BALB/c nude mice, 500µL 94µM ATN-224, delivered subcutaneously within the Matrigel plug significantly inhibited angiogenesis[4]. In nude mice bearing CAL27 squamous cell carcinoma (SCC) xenografts, daily oral gavage of 0.7mg ATN-224 combined with intratumoral Phosphate-Buffered Saline (PBS) or RAMBO on days 7 and 16 resulted in a significantly smaller average tumor volume in the RAMBO group by day 21 after ATN-224 treatment initiation[7].
References:
[1] Trapp V, Lee K, Doñate F, et al. Redox-related antimelanoma activity of ATN-224. Melanoma Res. 2009;19(6):350-360.
[2] Lowndes SA, Sheldon HV, Cai S, et al. Copper chelator ATN-224 inhibits endothelial function by multiple mechanisms. Microvasc Res. 2009;77(3):314-326.
[3] Doñate F, Juarez JC, Burnett ME, et al. Identification of biomarkers for the antiangiogenic and antitumour activity of the superoxide dismutase 1 (SOD1) inhibitor tetrathiomolybdate (ATN-224). Br J Cancer. 2008;98(4):776-783.
[4] Juarez JC, Betancourt O Jr, Pirie-Shepherd SR, et al. Copper binding by tetrathiomolybdate attenuates angiogenesis and tumor cell proliferation through the inhibition of superoxide dismutase 1. Clin Cancer Res. 2006;12(16):4974-4982.
[5] Lee K, Briehl MM, Mazar AP, et al. The copper chelator ATN-224 induces peroxynitrite-dependent cell death in hematological malignancies. Free Radic Biol Med. 2013;60:157-167.
[6] Lee K, Hart MR, Briehl MM, et al. The copper chelator ATN-224 induces caspase-independent cell death in diffuse large B-cell lymphoma. Int J Oncol. 2014;45(1):439-447.
[7] Yoo JY, Yu JG, Kaka A, et al. ATN-224 enhances antitumor efficacy of oncolytic herpes virus against both local and metastatic head and neck squamous cell carcinoma. Mol Ther Oncolytics. 2015;2:15008.
ATN-224是四硫代钼酸铵（tetrathiomolybdate, TM）的双胆碱盐，为超氧化物歧化酶抑制剂和高亲和力铜螯合剂，可抑制血液及内皮细胞中的CuZn超氧化物歧化酶1（SOD1）活性，从而发挥抗血管生成和抗肿瘤作用[1-3]。ATN-224以剂量（0、0.625、1.25、2.5、5、10μM）和时间（0、1、2、4、6、8、16、24、48h）依赖方式抑制成纤维细胞生长因子-2（FGF-2）诱导的细胞外信号调节激酶1/2（ERK1/2）磷酸化，其IC50介于1.25-2.5μM之间[4]。
在体外，0-10nM的ATN-224处理WEHI7.2及其衍生细胞（过氧化氢耐受的200R细胞和Bcl-2过表达的Hb12细胞）48h后，活细胞数呈浓度依赖性下降，EC50分别为3.17±0.27、5.84±0.34和5.25±0.32nM；同时，caspase-3活性与碘化丙啶（PI）摄取显著升高[5]。以32.71±2.04nM的ATN-224处理SUDHL-10细胞，12h后线粒体膜电位（ΔΨm）显著下降，48h后Mcl-1蛋白水平明显降低[6]。
在体内，在雌性BALB/c裸鼠Matrigel栓模型中，500μL 94μM的ATN-224皮下注射在Matriger塞内显著抑制血管生成[4]。在CAL27鳞状细胞癌（SCC）裸鼠移植瘤模型中，每日口服0.7mg的ATN-224，并于第7和第16天瘤内注射PBS或RAMBO，治疗开始后第21天RAMBO组的平均肿瘤体积显著小于对照组[7]。