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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author 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
author_facet 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
author_sort Lucas Rubini-Dias
collection DOAJ
description <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.
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spelling doaj-art-bbb8f9e3e025481b85b3ad332170a0d42025-08-20T02:00:55ZengMDPI AGBiomedicines2227-90592024-12-011212291610.3390/biomedicines12122916Mannich Base Derived from Lawsone Inhibits PKM2 and Induces Neoplastic Cell DeathLucas Rubini-Dias0Tácio V. A. Fernandes1Michele P. de Souza2Déborah Hottz3Afonso T. Arruda4Amanda de A. Borges5Gabriel Ouverney6Fernando de C. da Silva7Luana da S. M. Forezi8Gabriel Limaverde-Sousa9Bruno K. Robbs10Programa de Pós-Graduação em Ciências Morfológicas, Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Fundão, Rio de Janeiro 21941-590, RJ, BrazilDepartamento de Síntese de Fármacos, Instituto de Tecnologia em Fármacos, Farmanguinhos–Fiocruz, Manguinhos, Rio de Janeiro 21041-250, RJ, BrazilPostgraduate Program in Applied Science for Health Products, Faculty of Pharmacy, Fluminense Federal University, Niterói 24020-141, RJ, BrazilDepartamento de Ciência Básica, Instituto de Saúde de Nova Fribrugo, Universidade Federal Fluminense, Nova Friburgo 28625-650, RJ, BrazilPrograma de Pós-Graduação em Ciências Morfológicas, Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Fundão, Rio de Janeiro 21941-590, RJ, BrazilDepartamento de Química Orgânica, Instituto de Química, Campus do Valonguinho, Universidade Federal Fluminense, Niterói 24020-150, RJ, BrazilPrograma de Pós-Graduação em Ciências Morfológicas, Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Fundão, Rio de Janeiro 21941-590, RJ, BrazilDepartamento de Química Orgânica, Instituto de Química, Campus do Valonguinho, Universidade Federal Fluminense, Niterói 24020-150, RJ, BrazilDepartamento de Química Orgânica, Instituto de Química, Campus do Valonguinho, Universidade Federal Fluminense, Niterói 24020-150, RJ, BrazilInstituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro 21040-900, RJ, BrazilDepartamento de Ciência Básica, Instituto de Saúde de Nova Fribrugo, Universidade Federal Fluminense, Nova Friburgo 28625-650, RJ, Brazil<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.https://www.mdpi.com/2227-9059/12/12/2916oral squamous cell carcinomaMannich basesM2-type pyruvate kinaseautophagymolecular docking simulation
spellingShingle 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
Mannich Base Derived from Lawsone Inhibits PKM2 and Induces Neoplastic Cell Death
Biomedicines
oral squamous cell carcinoma
Mannich bases
M2-type pyruvate kinase
autophagy
molecular docking simulation
title Mannich Base Derived from Lawsone Inhibits PKM2 and Induces Neoplastic Cell Death
title_full Mannich Base Derived from Lawsone Inhibits PKM2 and Induces Neoplastic Cell Death
title_fullStr Mannich Base Derived from Lawsone Inhibits PKM2 and Induces Neoplastic Cell Death
title_full_unstemmed Mannich Base Derived from Lawsone Inhibits PKM2 and Induces Neoplastic Cell Death
title_short Mannich Base Derived from Lawsone Inhibits PKM2 and Induces Neoplastic Cell Death
title_sort mannich base derived from lawsone inhibits pkm2 and induces neoplastic cell death
topic oral squamous cell carcinoma
Mannich bases
M2-type pyruvate kinase
autophagy
molecular docking simulation
url https://www.mdpi.com/2227-9059/12/12/2916
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