Exploring the Antimycobacterial Potential of Podocarpusflavone A from <i>Kielmeyera membranacea</i>: In Vitro and In Vivo Insights

<b>Background/Objectives</b>: Tuberculosis (TB) is one of the leading infectious causes of death worldwide, highlighting the importance of identifying new anti-TB agents. In previous research, our team identified antimycobacterial activity in <i>Kielmeyera membranacea</i> lea...

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Main Authors: Marlon Heggdorne de Araujo, Salomé Muñoz Sánchez, Thatiana Lopes Biá Ventura Simão, Natalia Nowik, Stella Schuenck Antunes, Shaft Corrêa Pinto, Davide Sorze, Francesca Boldrin, Riccardo Manganelli, Nelilma Correia Romeiro, Elena B. Lasunskaia, Fons J. Verbeek, Herman P. Spaink, Michelle Frazão Muzitano
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Language:English
Published: MDPI AG 2024-11-01
Series:Pharmaceuticals
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Online Access:https://www.mdpi.com/1424-8247/17/12/1560
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author Marlon Heggdorne de Araujo
Salomé Muñoz Sánchez
Thatiana Lopes Biá Ventura Simão
Natalia Nowik
Stella Schuenck Antunes
Shaft Corrêa Pinto
Davide Sorze
Francesca Boldrin
Riccardo Manganelli
Nelilma Correia Romeiro
Elena B. Lasunskaia
Fons J. Verbeek
Herman P. Spaink
Michelle Frazão Muzitano
author_facet Marlon Heggdorne de Araujo
Salomé Muñoz Sánchez
Thatiana Lopes Biá Ventura Simão
Natalia Nowik
Stella Schuenck Antunes
Shaft Corrêa Pinto
Davide Sorze
Francesca Boldrin
Riccardo Manganelli
Nelilma Correia Romeiro
Elena B. Lasunskaia
Fons J. Verbeek
Herman P. Spaink
Michelle Frazão Muzitano
author_sort Marlon Heggdorne de Araujo
collection DOAJ
description <b>Background/Objectives</b>: Tuberculosis (TB) is one of the leading infectious causes of death worldwide, highlighting the importance of identifying new anti-TB agents. In previous research, our team identified antimycobacterial activity in <i>Kielmeyera membranacea</i> leaf extract; therefore, this study aims to conduct further exploration of its potential. <b>Methods</b>: Classical chromatography was applied for fractionation and spectrometric techniques were utilized for chemical characterization. For in vitro tests, samples were assessed against <i>Mycobacterium tuberculosis</i> and <i>Mycobacterium marinum</i>. The toxicity and efficacy of active samples were evaluated in vivo using different zebrafish models. Chemogenomics studies were applied to predict the isolated active compound’s potential mode of action. <b>Results</b>: We performed fractionation of <i>K. membranacea</i> ethanolic extract (EE) and then its dichloromethane fraction (DCM), and the biflavonoid podocarpusflavone A (PCFA) was isolated and identified as a promising active compound. The EE and PCFA were found to be non-toxic to zebrafish larvae and were able to inhibit <i>M. tuberculosis</i> growth extracellularly. Additionally, PCFA demonstrated antimycobacterial activity within infected macrophages, especially when combined with isoniazid. In addition, the EE, DCM, and PCFA have shown the ability to inhibit <i>M. marinum’s</i> growth during in vivo zebrafish larvae yolk infection. Notably, PCFA also effectively countered systemic infection established through the caudal vein, showing a similar inhibitory activity profile to rifampicin, both at 32 µM. A reduction in the transcriptional levels of pro-inflammatory cytokines confirmed the infection resolution. The protein tyrosine phosphatase B (PtpB) of <i>M</i>. <i>tuberculosis</i>, which inhibits the macrophage immune response, was predicted as a theoretical target of PCFA. This finding is in agreement with the higher activity observed for PCFA intracellularly and in vivo on zebrafish, compared with the direct action in <i>M. tuberculosis</i>. <b>Conclusions</b>: Here, we describe the discovery of PCFA as an intracellular inhibitor of <i>M. tuberculosis</i> and provide evidence of its in vivo efficacy and safety, encouraging its further development as a combination drug in novel therapeutic regimens for TB.
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spelling doaj-art-195b9fc75d8f42d0bbe63fdeabdcdeac2025-08-20T02:56:51ZengMDPI AGPharmaceuticals1424-82472024-11-011712156010.3390/ph17121560Exploring the Antimycobacterial Potential of Podocarpusflavone A from <i>Kielmeyera membranacea</i>: In Vitro and In Vivo InsightsMarlon Heggdorne de Araujo0Salomé Muñoz Sánchez1Thatiana Lopes Biá Ventura Simão2Natalia Nowik3Stella Schuenck Antunes4Shaft Corrêa Pinto5Davide Sorze6Francesca Boldrin7Riccardo Manganelli8Nelilma Correia Romeiro9Elena B. Lasunskaia10Fons J. Verbeek11Herman P. Spaink12Michelle Frazão Muzitano13Laboratório de Produtos Bioativos (LPBio), Instituto de Ciências Farmacêuticas, Universidade Federal do Rio de Janeiro, Campus Macaé, Macaé 27930-560, RJ, BrazilDepartment of Animal Sciences and Health, Institute of Biology (IBL), Leiden University, 2333 BE Leiden, The NetherlandsLaboratório de Biologia do Reconhecer (LBR), Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Campos dos Goytacazes 28013-602, RJ, BrazilDepartment of Animal Sciences and Health, Institute of Biology (IBL), Leiden University, 2333 BE Leiden, The NetherlandsLaboratório de Produtos Bioativos (LPBio), Instituto de Ciências Farmacêuticas, Universidade Federal do Rio de Janeiro, Campus Macaé, Macaé 27930-560, RJ, BrazilLaboratório de Produtos Bioativos (LPBio), Instituto de Ciências Farmacêuticas, Universidade Federal do Rio de Janeiro, Campus Macaé, Macaé 27930-560, RJ, BrazilDepartment of Molecular Medicine, University of Padova, 35121 Padova, ItalyDepartment of Molecular Medicine, University of Padova, 35121 Padova, ItalyDepartment of Molecular Medicine, University of Padova, 35121 Padova, ItalyLaboratório Integrado de Computação Científica (LICC), Universidade Federal do Rio de Janeiro, Campus Macaé, Macaé 27930-560, RJ, BrazilLaboratório de Biologia do Reconhecer (LBR), Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Campos dos Goytacazes 28013-602, RJ, BrazilLeiden Institute of Advanced Computer Science, Leiden University, 2333 CA Leiden, The NetherlandsDepartment of Animal Sciences and Health, Institute of Biology (IBL), Leiden University, 2333 BE Leiden, The NetherlandsLaboratório de Produtos Bioativos (LPBio), Instituto de Ciências Farmacêuticas, Universidade Federal do Rio de Janeiro, Campus Macaé, Macaé 27930-560, RJ, Brazil<b>Background/Objectives</b>: Tuberculosis (TB) is one of the leading infectious causes of death worldwide, highlighting the importance of identifying new anti-TB agents. In previous research, our team identified antimycobacterial activity in <i>Kielmeyera membranacea</i> leaf extract; therefore, this study aims to conduct further exploration of its potential. <b>Methods</b>: Classical chromatography was applied for fractionation and spectrometric techniques were utilized for chemical characterization. For in vitro tests, samples were assessed against <i>Mycobacterium tuberculosis</i> and <i>Mycobacterium marinum</i>. The toxicity and efficacy of active samples were evaluated in vivo using different zebrafish models. Chemogenomics studies were applied to predict the isolated active compound’s potential mode of action. <b>Results</b>: We performed fractionation of <i>K. membranacea</i> ethanolic extract (EE) and then its dichloromethane fraction (DCM), and the biflavonoid podocarpusflavone A (PCFA) was isolated and identified as a promising active compound. The EE and PCFA were found to be non-toxic to zebrafish larvae and were able to inhibit <i>M. tuberculosis</i> growth extracellularly. Additionally, PCFA demonstrated antimycobacterial activity within infected macrophages, especially when combined with isoniazid. In addition, the EE, DCM, and PCFA have shown the ability to inhibit <i>M. marinum’s</i> growth during in vivo zebrafish larvae yolk infection. Notably, PCFA also effectively countered systemic infection established through the caudal vein, showing a similar inhibitory activity profile to rifampicin, both at 32 µM. A reduction in the transcriptional levels of pro-inflammatory cytokines confirmed the infection resolution. The protein tyrosine phosphatase B (PtpB) of <i>M</i>. <i>tuberculosis</i>, which inhibits the macrophage immune response, was predicted as a theoretical target of PCFA. This finding is in agreement with the higher activity observed for PCFA intracellularly and in vivo on zebrafish, compared with the direct action in <i>M. tuberculosis</i>. <b>Conclusions</b>: Here, we describe the discovery of PCFA as an intracellular inhibitor of <i>M. tuberculosis</i> and provide evidence of its in vivo efficacy and safety, encouraging its further development as a combination drug in novel therapeutic regimens for TB.https://www.mdpi.com/1424-8247/17/12/1560biflavonepodocarpusflavone Azebrafishtuberculosis<i>Kielmeyera</i>cytokines
spellingShingle Marlon Heggdorne de Araujo
Salomé Muñoz Sánchez
Thatiana Lopes Biá Ventura Simão
Natalia Nowik
Stella Schuenck Antunes
Shaft Corrêa Pinto
Davide Sorze
Francesca Boldrin
Riccardo Manganelli
Nelilma Correia Romeiro
Elena B. Lasunskaia
Fons J. Verbeek
Herman P. Spaink
Michelle Frazão Muzitano
Exploring the Antimycobacterial Potential of Podocarpusflavone A from <i>Kielmeyera membranacea</i>: In Vitro and In Vivo Insights
Pharmaceuticals
biflavone
podocarpusflavone A
zebrafish
tuberculosis
<i>Kielmeyera</i>
cytokines
title Exploring the Antimycobacterial Potential of Podocarpusflavone A from <i>Kielmeyera membranacea</i>: In Vitro and In Vivo Insights
title_full Exploring the Antimycobacterial Potential of Podocarpusflavone A from <i>Kielmeyera membranacea</i>: In Vitro and In Vivo Insights
title_fullStr Exploring the Antimycobacterial Potential of Podocarpusflavone A from <i>Kielmeyera membranacea</i>: In Vitro and In Vivo Insights
title_full_unstemmed Exploring the Antimycobacterial Potential of Podocarpusflavone A from <i>Kielmeyera membranacea</i>: In Vitro and In Vivo Insights
title_short Exploring the Antimycobacterial Potential of Podocarpusflavone A from <i>Kielmeyera membranacea</i>: In Vitro and In Vivo Insights
title_sort exploring the antimycobacterial potential of podocarpusflavone a from i kielmeyera membranacea i in vitro and in vivo insights
topic biflavone
podocarpusflavone A
zebrafish
tuberculosis
<i>Kielmeyera</i>
cytokines
url https://www.mdpi.com/1424-8247/17/12/1560
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