Background: Currently, there is a significant lack of effective pharmacological agents for the treatment of breast cancer. Acylglycerol Kinase (AGK), a lipid kinase, has been found to be aberrantly expressed in breast cancer and is closely associated with tumor proliferation, migration, and invasion. However, no clinical anti-tumor drugs specifically targeting this kinase have been developed.Methods: siRNA was utilized to knock down the AGK gene; CCK8 and colony formation assays were employed to evaluate the in vitro proliferative capacity of tumor cells. Molecular dynamics simulations and BIL assays were conducted to analyze drug binding affinity. Annexin V/PI staining was used to assess apoptotic phenomena; subcutaneous xenograft tumor experiments in nude mice were performed to confirm the in vivo anti-tumor efficacy of the drug.Results: Netupitant exhibited stable binding affinity for AGK and interacted with amino acids within the ATP-binding region of the enzyme. The IC50values for the SK-BR-3 and MDA-MB-231 cell lines were determined as 16.15 ± 4.25 µmol/L and 24.02 ± 4.19 µmol/L, respectively; at a concentration of 2.5 µmol/L, Netupitant effectively inhibited clonogenic capacity in breast cancer cells; furthermore, treatment with 10 µmol/L significantly induced apoptosis in these cells. Doses of 50 mg/kg and 100 mg/kg Netupitant markedly suppressed growth rates of subcutaneous xenograft tumors in nude mice while also promoting apoptotic processes. Both in vivo and in vitro studies indicated that Netupitant could inhibit the activation of the PI3K/AKT/mTOR signaling pathway.Conclusions: By targeting AGK, Netupitant inhibits its kinase activity, which leads to reduced phosphorylation levels of PTEN, thereby suppressing the activation of the PI3K/AKT/mTOR signaling pathway and ultimately resulting in apoptosis in breast cancer cells.
背景：目前，针对乳腺癌的治疗仍缺乏有效的药物靶点。脂质激酶酰基甘油激酶（AGK）在乳腺癌中异常表达，与肿瘤增殖、迁移及侵袭密切相关，但尚未有靶向该激酶的临床抗肿瘤药物问世。 方法：利用siRNA敲低AGK基因；采用CCK8与克隆形成实验评估肿瘤细胞体外增殖能力；通过分子动力学模拟与BIL实验分析药物结合力；使用Annexin V/PI染色检测细胞凋亡现象；开展裸鼠皮下移植瘤实验验证药物体内抗肿瘤效果。 结果：奈妥匹坦与AGK具有稳定结合力，并与该酶ATP结合区氨基酸发生相互作用；其对SK-BR-3和MDA-MB-231细胞系的IC50值分别为16.15±4.25 µmol/L和24.02±4.19 µmol/L；2.5 µmol/L浓度可有效抑制乳腺癌细胞克隆形成能力；10 µmol/L处理能显著诱导细胞凋亡；50 mg/kg与100 mg/kg剂量的奈妥匹坦可明显抑制裸鼠皮下移植瘤生长速率并促进凋亡进程。体内外实验均表明奈妥匹坦能抑制PI3K/AKT/mTOR信号通路激活。 结论：奈妥匹坦通过靶向AGK抑制其激酶活性，降低PTEN磷酸化水平，进而抑制PI3K/AKT/mTOR信号通路激活，最终诱导乳腺癌细胞发生凋亡。
Netupitant Inhibits the Proliferation of Breast Cancer Cells by Targeting AGK
肿瘤(癌症)患者之家