Rubini-Dias Lucas, Fernandes Tácio V A, de Souza Michele P, Hottz Déborah, Arruda Afonso T, Borges Amanda de A, Ouverney Gabriel, da Silva Fernando de C, Forezi Luana da S M, Limaverde-Sousa Gabriel, Robbs Bruno K
Programa de Pós-Graduação em Ciências Morfológicas, Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Fundão, Rio de Janeiro 21941-590, RJ, Brazil.
Departamento de Síntese de Fármacos, Instituto de Tecnologia em Fármacos, Farmanguinhos-Fiocruz, Manguinhos, Rio de Janeiro 21041-250, RJ, Brazil.
Biomedicines. 2024 Dec 21;12(12):2916. doi: 10.3390/biomedicines12122916.
Pyruvate kinase M2, a central regulator of cancer cell metabolism, has garnered significant attention as a promising target for disrupting the metabolic adaptability of tumor cells. This study explores the potential of the Mannich base derived from lawsone () to interfere with PKM2 enzymatic activity both in vitro and in silico. The antiproliferative potential of was tested using MTT assay in various cell lines, including SCC-9, Hep-G2, HT-29, B16-F10, and normal human gingival fibroblast (HGF). The inhibition of PKM2 mediated by was assessed using an LDH-coupled assay and by measuring ATP production. Docking studies and molecular dynamics calculations were performed using Autodock 4 and GROMACS, respectively, on the tetrameric PKM2 crystallographic structure. The Mannich base demonstrated selective cytotoxicity against all cancer cell lines tested without affecting cell migration, with the highest selectivity index (SI) of 4.63 in SCC-9, followed by B16-F10 (SI = 3.9), Hep-G2 (SI = 3.4), and HT-29 (SI = 2.03). The compound effectively inhibited PKM2 glycolytic activity, leading to a reduction of ATP production both in the enzymatic reaction and in cells treated with this naphthoquinone derivative. showed favorable binding to PKM2 in the ATP-bound monomers through docking studies (PDB ID: 4FXF; binding affinity scores ranging from -6.94 to -9.79 kcal/mol) and MD simulations, revealing binding affinities stabilized by key interactions including hydrogen bonds, halogen bonds, and hydrophobic contacts. The findings suggest that exerts its antiproliferative activity by disrupting cell glucose metabolism, consequently reducing ATP production and triggering energetic collapse in cancer cells. This study highlights the potential of as a lead compound targeting PKM2 and warrants further investigation into its mechanism of action and potential clinical applications.
丙酮酸激酶M2是癌细胞代谢的核心调节因子，作为破坏肿瘤细胞代谢适应性的一个有前景的靶点已受到广泛关注。本研究探讨了源自胡桃醌的曼尼希碱（）在体外和计算机模拟中干扰丙酮酸激酶M2酶活性的潜力。使用MTT法在包括SCC - 9、Hep - G2、HT - 29、B16 - F10和正常人牙龈成纤维细胞（HGF）在内的多种细胞系中测试了的抗增殖潜力。使用LDH偶联测定法并通过测量ATP生成来评估介导的对丙酮酸激酶M2的抑制作用。分别使用Autodock 4和GROMACS对四聚体丙酮酸激酶M2晶体结构进行对接研究和分子动力学计算。曼尼希碱对所有测试的癌细胞系均表现出选择性细胞毒性，且不影响细胞迁移，在SCC - 9中选择性指数（SI）最高，为4.63，其次是B16 - F10（SI = 3.9）、Hep - G2（SI = 3.4）和HT - 29（SI = 2.03）。该化合物有效抑制丙酮酸激酶M2的糖酵解活性，导致酶促反应和用该萘醌衍生物处理的细胞中ATP生成减少。通过对接研究（PDB ID：4FXF；结合亲和力得分范围为 - 6.94至 - 9.79 kcal/mol）和分子动力学模拟显示，在ATP结合的单体中与丙酮酸激酶M2具有良好的结合，揭示了通过包括氢键、卤键和疏水接触在内的关键相互作用稳定的结合亲和力。研究结果表明，通过破坏细胞葡萄糖代谢发挥其抗增殖活性，从而减少ATP生成并引发癌细胞的能量崩溃。本研究突出了作为靶向丙酮酸激酶M2的先导化合物的潜力，值得进一步研究其作用机制和潜在的临床应用。
