A ferrocene-containing analogue of the MCU inhibitor Ru265 with increased cell permeability
A ferrocene-containing analogue of the MCU inhibitor Ru265 with increased cell permeability
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MCU 抑制剂 Ru265 的含二茂铁类似物，具有增加的细胞通透性
DOI：
10.1039/d2qi02183h
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发表时间：
2023
影响因子：
7
通讯作者：
Wilson, Justin J.
中科院分区：
文献类型：
--
作者：
Huang, Zhouyang;Spivey, Jesse A.;MacMillan, Samantha N.;Wilson, Justin J.
线粒体钙单转运蛋白（MCU）是一种介导线粒体钙（mCa2+）摄取的跨膜蛋白。MCU抑制剂作为研究mCa2+摄取对细胞功能的作用的工具而备受关注。在这项研究中，我们报道了两种有效的MCU抑制剂，[Ru2(μ-N)(NH3)8(FcCO2)2](OTf)3 （RuOFc, Fc =二茂铁，OTf =三氟酸盐）和[Ru2(μ-N)(NH3)8(PhCO2)2](OTf)3 （RuOBz）。这些化合物是先前报道的抑制剂[Ru2(μ-N)(NH3)8(Cl)2](Cl)3 （Ru265）的类似物，它们分别与二茂铁羧酸酯和苯甲酸酯配体衍生。合成了两种化合物，并用核磁共振光谱、红外光谱和x射线晶体学对其进行了表征。在生理条件下，RuOFc和RuOBz水解生成[Ru2(μ-N)(NH3)8(H2O)2](OTf)5 （Ru265 '）和游离羧酸盐，半衰期分别为2.9和6.5 h。若ofc在N，N ' -二甲基甲酰胺（DMF）中的循环伏安法显示，与SCE相比，在0.64 V时发生了显著的可逆2e -转移事件，对应于两种含二茂铁的轴向配体同时氧化。与SCE相比，这三种配合物在低于- 1 V的电位下也表现出不可逆的ru基还原。Ru265′、RuOFc和RuOBz的DFT计算证实，RuOFc的氧化还原活性来源于二茂铁配体。此外，三种化合物的LUMO能量与其不可逆还原势相关。对Ru265、RuOFc和RuOBz的生物学特性进行了系统的比较。RuOFc和RuOBz均能抑制HEK293T细胞对mCa2+的摄取，但其抑制作用比Ru265弱5-7倍。在完整细胞中，RuOBz被细胞吸收，抑制MCU的程度与Ru265相似。然而，RuOFc的细胞摄取比Ru265增加了10倍，这反过来也导致完整细胞中mcu抑制活性的适度增强。此外，与Ru265相比，RuOFc对HEK293T和HeLa细胞具有细胞毒性，其50%的生长抑制浓度分别为23.2 μM和33.9 μM，这一特性可用于开发靶向mcu的抗癌药物。这些结果表明RuOFc是一种有效的MCU抑制剂，也是Ru265的轴向配体功能化如何导致具有不同物理和生物特性的该类新化合物的另一个例子。
The mitochondrial calcium uniporter (MCU) is a transmembrane protein that mediates mitochondrial calcium (mCa2+) uptake. Inhibitors of the MCU are of interest for their applications as tools to study the role of mCa2+ uptake on cellular function. In this study, we report two potent MCU inhibitors, [Ru2(μ-N)(NH3)8(FcCO2)2](OTf)3 (RuOFc, Fc = ferrocene, OTf = triflate) and [Ru2(μ-N)(NH3)8(PhCO2)2](OTf)3 (RuOBz). These compounds are analogues of the previously reported inhibitor [Ru2(μ-N)(NH3)8(Cl)2](Cl)3 (Ru265) that has been derivatized with ferrocenecarboxylate and benzoate ligands, respectively. Both compounds were synthesized and fully characterized by NMR spectroscopy, infrared spectroscopy and X-ray crystallography. Under physiological conditions, RuOFc and RuOBz aquate with half-lives of 2.9 and 6.5 h, respectively, to produce [Ru2(μ-N)(NH3)8(H2O)2](OTf)5 (Ru265′) and the free carboxylates. Cyclic voltammetry of RuOFc in N,N′-dimethylformamide (DMF) reveals a prominent reversible 2e− transfer event at 0.64 V vs. SCE, corresponding to the simultaneous oxidation of both ferrocene-containing axial ligands. All three complexes also exhibit irreversible Ru-based reductions at potentials below −1 V vs. SCE. DFT calculations of Ru265′, RuOFc and RuOBz confirm that the redox activity of RuOFc arises from the ferrocene ligands. Furthermore, LUMO energies of the three compounds correlate with their irreversible reduction potentials. A systematic comparison on the biological properties of Ru265, RuOFc and RuOBz was carried out. Both RuOFc and RuOBz inhibit mCa2+ uptake in permeabilized HEK293T cells, but are 5–7 fold less potent than Ru265. In intact cells, RuOBz is taken up by cells and inhibits the MCU to a similar extent as Ru265. RuOFc, however, exhibits a 10-fold increase in cellular uptake over Ru265, which in turn also leads to a modest enhancement in MCU-inhibitory activity in intact cells. Moreover, in contrast to Ru265, RuOFc is cytotoxic to HEK293T and HeLa cells with 50% growth inhibitory concentration values of 23.2 and 33.9 μM, respectively, a property that could be leveraged to develop MCU-targeting anticancer agents. These results establish RuOFc as a potent MCU inhibitor and another example of how axial ligand functionalization of Ru265 can lead to new compounds within this class with diverse physical and biological properties.
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