组蛋白脱乙酰酶抑制剂 (HDACi) 贝利司他由于代谢稳定性差，对结肠癌等实体瘤的治疗作用有限。在这里，我们在体外和离体评估了一种新型贝利司他前药铜-双贝利司他 (Cubisbel)，旨在克服贝利司他的药代动力学挑战。使用质量数在人肝微粒体 (HLM) 中评估了每种 HDACi 的体外代谢光谱测定法。接下来，在三种结肠癌细胞系中评估了 belinostat 和 Cubisbel 对细胞生长、HDAC 活性、细胞凋亡和细胞周期的影响。使用 RNA-Seq 确定两种 HDAC 诱导的基因表达改变，然后进行计算机分析以确定差异表达基因 (DEG) 的主调节因子 (MR)。还检查了两种 HDACis 对结肠癌患者来源的肿瘤类器官 (PDTO) 活力的影响。Belinostat 和 Cubisbel 通过 HDAC 抑制和凋亡诱导显着降低了结肠癌细胞的生长。有趣的是，Cubisbel 的体外半衰期明显长于 belinostat。 Belinostat 及其 Cu 衍生物通常会失调许多信号传导和代谢途径，而 Cubisbel 下调的基因可能受到 VEGFA、ERBB2 和 DUSP2 MR 的控制。用 HDACis 治疗结肠癌 PDTO 导致细胞活力显着降低以及干细胞和增殖标记物的下调。 belinostat 与 Cu(II) 的复合不会改变 belinostat 的 HDAC 活性，而是显着增强其代谢稳定性。体外并通过扰乱结肠癌中的关键 MR 来靶向抗癌途径。 HDACis 与金属离子的络合可能会提高临床上使用的 HDACis 对结肠癌患者的疗效。© 2023。作者。

The histone deacetylase inhibitor (HDACi), belinostat, has had limited therapeutic impact in solid tumors, such as colon cancer, due to its poor metabolic stability. Here we evaluated a novel belinostat prodrug, copper-bis-belinostat (Cubisbel), in vitro and ex vivo, designed to overcome the pharmacokinetic challenges of belinostat.The in vitro metabolism of each HDACi was evaluated in human liver microsomes (HLMs) using mass spectrometry. Next, the effect of belinostat and Cubisbel on cell growth, HDAC activity, apoptosis and cell cycle was assessed in three colon cancer cell lines. Gene expression alterations induced by both HDACis were determined using RNA-Seq, followed by in silico analysis to identify master regulators (MRs) of differentially expressed genes (DEGs). The effect of both HDACis on the viability of colon cancer patient-derived tumor organoids (PDTOs) was also examined.Belinostat and Cubisbel significantly reduced colon cancer cell growth mediated through HDAC inhibition and apoptosis induction. Interestingly, the in vitro half-life of Cubisbel was significantly longer than belinostat. Belinostat and its Cu derivative commonly dysregulated numerous signalling and metabolic pathways while genes downregulated by Cubisbel were potentially controlled by VEGFA, ERBB2 and DUSP2 MRs. Treatment of colon cancer PDTOs with the HDACis resulted in a significant reduction in cell viability and downregulation of stem cell and proliferation markers.Complexation of belinostat to Cu(II) does not alter the HDAC activity of belinostat, but instead significantly enhances its metabolic stability in vitro and targets anti-cancer pathways by perturbing key MRs in colon cancer. Complexation of HDACis to a metal ion might improve the efficacy of clinically used HDACis in patients with colon cancer.© 2023. The Author(s).