Mannich Base Derived from Lawsone Inhibits PKM2 and Induces Neoplastic Cell Death

<b>Background/Objectives:</b> 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 (...

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Main Authors: Lucas Rubini-Dias, Tácio V. A. Fernandes, Michele P. de Souza, Déborah Hottz, Afonso T. Arruda, Amanda de A. Borges, Gabriel Ouverney, Fernando de C. da Silva, Luana da S. M. Forezi, Gabriel Limaverde-Sousa, Bruno K. Robbs
Format: Article
Language:English
Published: MDPI AG 2024-12-01
Series:Biomedicines
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Online Access:https://www.mdpi.com/2227-9059/12/12/2916
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Summary:<b>Background/Objectives:</b> 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 (<b>MB-6a</b>) to interfere with PKM2 enzymatic activity both in vitro and in silico. <b>Methods:</b> The antiproliferative potential of <b>MB-6a</b> 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 <b>MB-6a</b> 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. <b>Results:</b> The Mannich base <b>6a</b> 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. <b>MB-6a</b> 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. <b>Conclusions:</b> The findings suggest that <b>MB-6a</b> 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 <b>MB-6a</b> as a lead compound targeting PKM2 and warrants further investigation into its mechanism of action and potential clinical applications.
ISSN:2227-9059