BackgroundGP‐2250, a novel analog of taurultam (TRLT), has emerged as a potent anti‐neoplastic drug; however, the mechanisms underlying its effects are not well understood. Here, we investigated the mechanism of action and the biological effects of GP‐2250 using in vitro and in vivo models.MethodsWe carried out a series of in vitro (MTT assay, Annexin V/PI assay, colony formation assay, reverse‐phase protein array [RPPA], and HRLC/IC analysis) to determine the biological activity of GP‐2250 and investigate the mechanism of action. In vivo experiments were carried out to determine the therapeutic efficacy of GP‐2250 alone and in combination with standard‐of‐care drugs (e.g., paclitaxel, cisplatin, topotecan, and poly ADP‐ribose polymerase [PARP] inhibitors).ResultsWe investigated the cytotoxic effect of GP‐2250 in 10 ovarian cancer cell lines and found GP‐2250 combined with a PARP inhibitor had the greatest synergy. RPPA revealed that GP‐2250 inhibited hypoxia‐inducible factor‐1α, AKT, and mammalian target of rapamycin (mTOR) activation and expression. High‐resolution mass spectrometry revealed that hexokinase2 activity and protein expression were significantly reduced by GP‐2250 exposure. Furthermore, GP‐2250 reduced glycolysis and ATP synthesis in cancer cells. An in vivo pharmacodynamic experiment using the OVCAR8 mouse model demonstrated that 500 mg/kg GP‐2250 was effective in downregulating AKT and mTOR activation and expression. In the in vivo therapy experiment using an orthotopic mouse model, a combination of GP‐2250 with either PARP inhibitors or bevacizumab showed a significant reduction of tumor weights and nodules compared to those treated with a vehicle, control IgG groups, or monotherapy groups.ConclusionsTaken together, our data indicate that GP‐2250 exerts profound effects on tumor metabolism and, in combination with PARP inhibitors or bevacizumab, showed promising anti‐tumor efficacy. These findings could have implications for the clinical development of GP‐2250.

中文翻译：

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新型恶噻嗪衍生物GP-2250治疗卵巢癌的作用机制及合理组合


背景GP-2250是一种新型的taurultam类似物（TRLT），已成为一种有效的抗肿瘤药物；然而，其影响的机制尚不清楚。在此，我们利用体外和体内模型研究了 GP-2250 的作用机制和生物学效应。方法我们进行了一系列体外实验（MTT 测定、Annexin V/PI 测定、集落形成测定、反相蛋白阵列[RPPA]和HRLC/IC分析）来确定GP-2250的生物活性并研究作用机制。进行体内实验以确定 GP-2250 单独使用以及与标准治疗药物（例如紫杉醇、顺铂、托泊替康和聚 ADP-核糖聚合酶 [PARP] 抑制剂）联合使用的治疗效果。 GP-2250 在 10 种卵巢癌细胞系中的细胞毒作用，发现 GP-2250 与 PARP 抑制剂联合具有最大的协同作用。 RPPA 显示 GP-2250 抑制缺氧诱导因子 1α、AKT 和哺乳动物雷帕霉素靶点 (mTOR) 的激活和表达。高分辨率质谱分析显示，接触 GP-2250 后，己糖激酶 2 活性和蛋白质表达显着降低。此外，GP-2250 减少癌细胞中的糖酵解和 ATP 合成。使用 OVCAR8 小鼠模型进行的体内药效学实验表明，500 mg/kg GP-2250 可有效下调 AKT 和 mTOR 激活和表达。在使用原位小鼠模型的体内治疗实验中，与媒介物、对照 IgG 组或单一治疗组相比，GP-2250 与 PARP 抑制剂或贝伐单抗的组合显示肿瘤重量和结节显着减少。结论综上所述，我们的数据表明 GP-2250 对肿瘤代谢产生深远影响，并且与 PARP 抑制剂或贝伐珠单抗联合使用，显示出有希望的抗肿瘤功效。这些发现可能对 GP-2250 的临床开发产生影响。