Facultad de Ciencias Químicas, Benemérita Universidad Autónoma de Puebla, Ciudad Universitaria, 72570 Puebla, PUE, Mexico; Departamento de Química Orgánica, Facultad de Química, Universidad de Sevilla, Apartado 1203, E-41071 Seville, Spain.
BioLab, Instituto Universitario de Bio-Orgánica "Antonio González", Universidad de la Laguna, C/ Astrofísico Francisco Sánchez 2, 38206 La Laguna, Spain.
Bioorg Chem. 2024 Apr;145:107168. doi: 10.1016/j.bioorg.2024.107168. Epub 2024 Feb 6.
Being aware of the need to develop more efficient therapies against cancer, herein we disclose an innovative approach for the design of selective antiproliferative agents. We have accomplished the conjugation of a coumarin fragment with lipophilic cations (triphenylphosphonium salts, guanidinium) for providing mitochondriotropic agents that simultaneously target also carbonic anhydrases IX and XII, involved in the development and progression of cancer. The new compounds prepared herein turned out to be strong inhibitors of carbonic anhydrases IX and XII of human origin (low-to-mid nM range), also endowed with high selectivity, exhibiting negligible activity towards cytosolic CA isoforms. Key interactions with the enzyme were analysed using docking and molecular dynamics simulations. Regarding their in vitro antiproliferative activities, an increase of the tether length connecting both pharmacophores led to a clear improvement in potency, reaching the submicromolar range for the lead compounds, and an outstanding selectivity towards tumour cell lines (S.I. up to >357). Cytotoxic effects were also analysed on MDR cell lines under hypoxic and normoxic conditions. Chemoresistance exhibited by phosphonium salts, and not by guanidines, against MDR cells was based on the fact that the former were found to be substrates of P-glycoprotein (P-gp), the pump responsible for extruding foreign chemicals; this situation was reversed by administrating tariquidar, a third generation P-gp inhibitor. Moreover, phosphonium salts provoked a profound depolarization of mitochondria membranes from tumour cells, thus probably compromising their oxidative metabolism. To gain insight into the mode of action of title compounds, continuous live cell microscopy was employed; interestingly, this technique revealed two different antiproliferative mechanisms for both families of mitocans. Whereas phosphonium salts had a cytostatic effect, blocking cell division, guanidines led to cell death via apoptosis.
意识到需要开发更有效的癌症治疗方法，我们在此披露了一种设计选择性抗增殖剂的创新方法。我们已经完成了香豆素片段与亲脂阳离子（三苯基膦盐、胍盐）的连接，以提供同时靶向碳酸酐酶 IX 和 XII 的促线粒体剂，这些酶参与癌症的发展和进展。本文制备的新化合物被证明是强效的人源碳酸酐酶 IX 和 XII 抑制剂（低至中 nM 范围），同时具有高选择性，对细胞溶质 CA 同工酶表现出可忽略的活性。使用对接和分子动力学模拟分析了与酶的关键相互作用。关于它们的体外抗增殖活性，连接两个药效团的连接体长度的增加导致活性明显提高，达到了先导化合物的亚微摩尔范围，并且对肿瘤细胞系具有出色的选择性（S.I.高达>357）。还在缺氧和常氧条件下分析了对 MDR 细胞系的细胞毒性作用。磷盐而非胍盐对 MDR 细胞的化学抗性基于这样一个事实，即前者被发现是 P-糖蛋白（P-gp）的底物，P-gp 是负责排出外来化学物质的泵；通过给予第三代 P-gp 抑制剂 tariquidar，可以逆转这种情况。此外，磷盐会引起肿瘤细胞线粒体膜的严重去极化，从而可能破坏其氧化代谢。为了深入了解标题化合物的作用方式，我们采用了连续活细胞显微镜技术；有趣的是，这项技术揭示了两种不同的促线粒体剂家族的两种不同的抗增殖机制。磷盐具有细胞停滞作用，阻止细胞分裂，而胍盐则通过细胞凋亡导致细胞死亡。
