Búsqueda de inhibidores del transportador de membrana CtpF de Mycobacterium tuberculosis mediante screening virtual basado en farmacóforo y determinación in vitro de la actividad
ilustraciones, gráficas
- Autores:
-
Varón Vega, Henry Andrés
- Tipo de recurso:
- Fecha de publicación:
- 2022
- Institución:
- Universidad Nacional de Colombia
- Repositorio:
- Universidad Nacional de Colombia
- Idioma:
- spa
- OAI Identifier:
- oai:repositorio.unal.edu.co:unal/82682
- Palabra clave:
- 540 - Química y ciencias afines
Mycobacterium
Efecto de los fármacos
Inmunología
Drug effects
Immunology
Tuberculosis
Fármaco-resistencia
ATPasas tipo P
acoplamiento molecular
Cribado virtual
CtpF
- Rights
- openAccess
- License
- Reconocimiento 4.0 Internacional
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Universidad Nacional de Colombia |
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| dc.title.spa.fl_str_mv |
Búsqueda de inhibidores del transportador de membrana CtpF de Mycobacterium tuberculosis mediante screening virtual basado en farmacóforo y determinación in vitro de la actividad |
| dc.title.translated.eng.fl_str_mv |
Search for Mycobacterium tuberculosis membrane transporter CtpF inhibitors by pharmacophore-based virtual screening and in vitro activity determination |
| title |
Búsqueda de inhibidores del transportador de membrana CtpF de Mycobacterium tuberculosis mediante screening virtual basado en farmacóforo y determinación in vitro de la actividad |
| spellingShingle |
Búsqueda de inhibidores del transportador de membrana CtpF de Mycobacterium tuberculosis mediante screening virtual basado en farmacóforo y determinación in vitro de la actividad 540 - Química y ciencias afines Mycobacterium Efecto de los fármacos Inmunología Drug effects Immunology Tuberculosis Fármaco-resistencia ATPasas tipo P acoplamiento molecular Cribado virtual CtpF |
| title_short |
Búsqueda de inhibidores del transportador de membrana CtpF de Mycobacterium tuberculosis mediante screening virtual basado en farmacóforo y determinación in vitro de la actividad |
| title_full |
Búsqueda de inhibidores del transportador de membrana CtpF de Mycobacterium tuberculosis mediante screening virtual basado en farmacóforo y determinación in vitro de la actividad |
| title_fullStr |
Búsqueda de inhibidores del transportador de membrana CtpF de Mycobacterium tuberculosis mediante screening virtual basado en farmacóforo y determinación in vitro de la actividad |
| title_full_unstemmed |
Búsqueda de inhibidores del transportador de membrana CtpF de Mycobacterium tuberculosis mediante screening virtual basado en farmacóforo y determinación in vitro de la actividad |
| title_sort |
Búsqueda de inhibidores del transportador de membrana CtpF de Mycobacterium tuberculosis mediante screening virtual basado en farmacóforo y determinación in vitro de la actividad |
| dc.creator.fl_str_mv |
Varón Vega, Henry Andrés |
| dc.contributor.advisor.none.fl_str_mv |
Soto Ospina, Carlos Yesid López Vallejo, Fabián Harvey |
| dc.contributor.author.none.fl_str_mv |
Varón Vega, Henry Andrés |
| dc.contributor.researchgroup.spa.fl_str_mv |
Bioquímica y Biología Molecular de las Micobacterias |
| dc.subject.ddc.spa.fl_str_mv |
540 - Química y ciencias afines |
| topic |
540 - Química y ciencias afines Mycobacterium Efecto de los fármacos Inmunología Drug effects Immunology Tuberculosis Fármaco-resistencia ATPasas tipo P acoplamiento molecular Cribado virtual CtpF |
| dc.subject.decs.spa.fl_str_mv |
Mycobacterium Efecto de los fármacos Inmunología |
| dc.subject.decs.eng.fl_str_mv |
Drug effects |
| dc.subject.lemb.eng.fl_str_mv |
Immunology |
| dc.subject.proposal.spa.fl_str_mv |
Tuberculosis Fármaco-resistencia ATPasas tipo P acoplamiento molecular Cribado virtual CtpF |
| description |
ilustraciones, gráficas |
| publishDate |
2022 |
| dc.date.accessioned.none.fl_str_mv |
2022-11-10T20:03:28Z |
| dc.date.available.none.fl_str_mv |
2022-11-10T20:03:28Z |
| dc.date.issued.none.fl_str_mv |
2022-09-13 |
| dc.type.spa.fl_str_mv |
Trabajo de grado - Maestría |
| dc.type.driver.spa.fl_str_mv |
info:eu-repo/semantics/masterThesis |
| dc.type.version.spa.fl_str_mv |
info:eu-repo/semantics/acceptedVersion |
| dc.type.content.spa.fl_str_mv |
Text |
| dc.type.redcol.spa.fl_str_mv |
http://purl.org/redcol/resource_type/TM |
| status_str |
acceptedVersion |
| dc.identifier.uri.none.fl_str_mv |
https://repositorio.unal.edu.co/handle/unal/82682 |
| dc.identifier.instname.spa.fl_str_mv |
Universidad Nacional de Colombia |
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Repositorio Institucional Universidad Nacional de Colombia |
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https://repositorio.unal.edu.co/ |
| url |
https://repositorio.unal.edu.co/handle/unal/82682 https://repositorio.unal.edu.co/ |
| identifier_str_mv |
Universidad Nacional de Colombia Repositorio Institucional Universidad Nacional de Colombia |
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spa |
| language |
spa |
| dc.relation.indexed.spa.fl_str_mv |
Bireme RedCol LaReferencia |
| dc.relation.references.spa.fl_str_mv |
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The bacillary and macrophage response to hypoxia in tuberculosis and the consequences for T cell antigen recognition. Microbes Infect. 19, 177–192 (2017). Novoa-Aponte, L. et al. In silico identification and characterization of the ion transport specificity for P-type ATPases in the Mycobacterium tuberculosis complex. BMC Struct. Biol. 12, 1–12 (2012). Ward, S. K., Abomoelak, B., Hoye, E. A., Steinberg, H. & Talaat, A. M. CtpV: a putative copper exporter required for full virulence of Mycobacterium tuberculosis. Mol. Microbiol. 77, 1096–1110 (2010). Raimunda, D., Long, J. E., Padilla-Benavides, T., Sassetti, C. M. & Argüello, J. M. Differential roles for the Co2+/Ni2+ transporting ATPases , CtpD and CtpJ , in Mycobacterium tuberculosis virulence. Mol. Microbiol. 91, 185–197 (2014). Maya-Hoyos, M., Rosales, C., Novoa-Aponte, L., Castillo, E. & Soto, C. Y. The P-type ATPase CtpF is a plasma membrane transporter mediating calcium efflux in Mycobacterium tuberculosis cells. 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Antibacterial and Hemolytic Activity of a new Lectin Purified from the Seeds of Sterculia Foetida L. Appl. Biochem. Biotechnol. 175, 1689–1699 (2015). Di Marino, D., D’Annessa, I., Coletta, A., Via, A. & Tramontano, A. Characterization of the differences in the cyclopiazonic acid binding mode to mammalian and P. Falciparum Ca2+ pumps: A computational study. Proteins Struct. Funct. Bioinforma. 83, 564–574 (2015). Rodríguez, Y. & Májeková, M. Structural Changes of Sarco/Endoplasmic Reticulum Ca2+-ATPase Induced by Rutin Arachidonate: A Molecular Dynamics Study. Biomolecules 10, (2020). Sahakyan, H. Improving virtual screening results with MM/GBSA and MM/PBSA rescoring. J. Comput. Aided. Mol. Des. 35, 731–736 (2021). Gilson, M. K. & Zhou, H.-X. Calculation of protein-ligand binding affinities. Annu. Rev. Biophys. Biomol. Struct. 36, 21–42 (2007). Aparna, V., Dineshkumar, K., Mohanalakshmi, N., Velmurugan, D. & Hopper, W. 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Toxicology 252, 72–77 (2008). Ge, F. et al. In vitro synergistic interactions of oleanolic acid in combination with isoniazid, rifampicin or ethambutol against Mycobacterium tuberculosis. J. Med. Microbiol. 59, 567–572 (2010). Mohyeldin, S. M., Mehanna, M. M. & Elgindy, N. A. The relevancy of controlled nanocrystallization on rifampicin characteristics and cytotoxicity. Int. J. Nanomedicine 11, 2209–2222 (2016). Smitha, K. T., Nisha, N., Maya, S., Biswas, R. & Jayakumar, R. Delivery of rifampicin-chitin nanoparticles into the intracellular compartment of polymorphonuclear leukocytes. Int. J. Biol. Macromol. 74, 36–43 (2015). International Standard Organization. ISO 10993-5:2009 Biological evaluation of medical devices — Part 5: Tests for in vitro cytotoxicity. 34 (2009). Clara, A.-P. et al. Synergy between Circular Bacteriocin AS-48 and Ethambutol against Mycobacterium tuberculosis. Antimicrob. Agents Chemother. 62, (2022). |
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Universidad Nacional de Colombia |
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Facultad de Ciencias |
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Reconocimiento 4.0 Internacionalhttp://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccesshttp://purl.org/coar/access_right/c_abf2Soto Ospina, Carlos Yesida5b5bbad5c61bb4a64f522a1f05590adLópez Vallejo, Fabián Harveyded3062059fd87d5db691f0ac84dde83Varón Vega, Henry Andrés3abea2eb4098cf0c003acce5c2dc560aBioquímica y Biología Molecular de las Micobacterias2022-11-10T20:03:28Z2022-11-10T20:03:28Z2022-09-13https://repositorio.unal.edu.co/handle/unal/82682Universidad Nacional de ColombiaRepositorio Institucional Universidad Nacional de Colombiahttps://repositorio.unal.edu.co/ilustraciones, gráficasLa aparición de cepas de Mycobacterium tuberculosis (Mtb), multirresistente y extremadamente resistente a los fármacos, y la coinfección con el virus de inmunodeficiencia humana (VIH), han aumentado dramáticamente la mortalidad por tuberculosis (TB) en el mundo. Esta situación obliga a la necesidad de encontrar dianas terapéuticas alternativas y la identificación de nuevos fármacos más efectivos contra la TB. En este sentido, las proteínas de membrana ATPasas tipo P, podrían ser blancos relevantes, toda vez que se conoce son relevantes en la homeostasis iónica y en la supervivencia de Mtb durante la infección. CtpF, una Ca2+-ATPasa de membrana plasmática, es una proteína que se activa en condiciones de hipoxia, estrés oxidativo e infección, sugiriendo qué este transportador hace parte de las estrategias usadas por el bacilo tuberculoso para evadir la respuesta inmune. Considerando la importancia de CtpF en la viabilidad del bacilo tuberculoso, en el presente trabajo se planteó la búsqueda de nuevos compuestos con potencial actividad inhibitoria del transporte de Ca2+ mediado por CtpF. En el presente trabajo se partió del análisis in silico de los modos de unión y las interacciones con los residuos funcionalmente activos entre CtpF y el ácido ciclopiazónico (CPA, un reconocido inhibidor de la actividad Ca2+ ATPasa en eucariotas) además de tres compuestos adicionales que en un estudio previo mostraron inhibición del crecimiento de Mtb y de la actividad ATPasa mediada por CtpF. Para la identificación de los nuevos inhibidores se llevó a cabo un cribado virtual basado en diferentes métodos quimioinformáticos que incluyeron: dinámica molecular, modelado del farmacóforo, huellas digitales moleculares, acoplamiento molecular automatizado y cálculos de energía de libre de unión. La estrategia propuesta en este trabajo permitió identificar once compuestos con potencial actividad inhibitoria de CtpF, los que se probaron in vitro. Ensayos de concentración mínima inhibitoria (CMI) permitieron seleccionar cuatro nuevos inhibidores del crecimiento de Mtb (ZINC57418826, ZINC12547355, ZINC09731847 y ZINC04030361). Experimentos para determinar el efecto de dos de estos compuestos sobre la actividad Ca2+-ATPasa de vesículas de membrana, mostraron que los ligandos ZINC09731847 (IC50 = 7,6 μM) y ZINC04030361 (IC50 = 3,3 μM) inhiben de manera efectiva la actividad de CtpF en membrana plasmática. Los resultados de CMI (25,0 μg/mL) actividad Ca2+-ATPasa, actividad citotóxica (27,2, %) sobre células vero y hemólisis sobre glóbulos rojos humanos (0,2 %), ayudaron a seleccionar el compuesto ZINC04030361 como el candidato más activo contra la ATPasa CtpF de membrana plasmática. Los resultados obtenidos en el presente trabajo representan un avance en la búsqueda y desarrollo de nuevos compuestos anti-TB dirigidos a dianas alternativas, como son las proteínas de membrana involucradas en la homeóstasis iónica de Mtb. (Texto tomado de la fuente)The appearance of multidrug-resistant and extremely drug-resistant Mycobacterium tuberculosis (Mtb) strains, and co-infection with the human immunodeficiency virus (HIV), have dramatically increased mortality from tuberculosis (TB) in the world. This situation forces the need to find alternative therapeutic targets and the identification of new and more effective drugs against TB. In this way, P-type ATPase membrane proteins are known to be relevant in ionic homeostasis and in the survival of Mtb during infection. CtpF, a Ca2+-ATPase, is a protein whose encoding gene is activated under conditions of hypoxia, oxidative stress, and infection, suggesting that this transporter is part of the strategies used by the tubercle bacillus to evade the immune response. Considering the importance of CtpF in the viability of the tubercle bacillus, was proposed in the present work the search for new compounds with potential inhibitory activity of Ca2+ transport mediated by CtpF. In the present work, we started from the in silico analysis of the binding modes and the interactions with the functionally active residues between CtpF and cyclopiazonic acid (CPA, a recognized inhibitor of Ca2+ ATPase activity in eukaryotes) and three additional compounds, which in a previous study showed inhibition of Mtb growth, as well as of CtpF mediated ATPase activity. For the identification of the new inhibitors, a virtual screening was carried out based on different chemoinformatic methods that included: molecular dynamics, pharmacophore modeling, molecular fingerprinting, automated molecular docking, and binding free energy calculations. The strategy proposed in this work allowed the identification of eleven compounds with potential CtpF inhibitory activity, these compounds were tested in vitro. Minimum inhibitory concentration (MIC) assays allowed the selection of four new Mtb growth inhibitors (ZINC57418826, ZINC12547355, ZINC09731847 and ZINC04030361). Experiments to determine the effect of two of these compounds on the Ca2+-ATPase activity of membrane vesicles, showed that the ligands ZINC09731847 (IC50 = 7,6 μM) and ZINC04030361 (IC50 = 3,3 μM) effectively inhibit CtpF activity. The results of MIC (25,0 μg/mL) Ca2+-ATPase activity, in addition to cytotoxic activity assays on Vero cells (27,2 %) and hemolysis on human red blood cells (0,2%), helped to select the compound ZINC04030361 as the most active candidate against the plasma membrane ATPase CtpF. 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Agents Chemother. 62, (2022).Bioquímica y Biología Molecular de las MicobacteriasEstudiantesInvestigadoresMaestrosPúblico generalLICENSElicense.txtlicense.txttext/plain; charset=utf-85879https://repositorio.unal.edu.co/bitstream/unal/82682/1/license.txteb34b1cf90b7e1103fc9dfd26be24b4aMD51ORIGINAL1118558029.2022.pdf1118558029.2022.pdfTesis de Maestría en Ciencias - Bioquímicaapplication/pdf2007232https://repositorio.unal.edu.co/bitstream/unal/82682/2/1118558029.2022.pdfb5b6135f1fef2b09e8c3ed47c68c4ee2MD52THUMBNAIL1118558029.2022.pdf.jpg1118558029.2022.pdf.jpgGenerated Thumbnailimage/jpeg4672https://repositorio.unal.edu.co/bitstream/unal/82682/3/1118558029.2022.pdf.jpg384b8d42cf741d4c6a16855dbbd14d6dMD53unal/82682oai:repositorio.unal.edu.co:unal/826822023-08-10 23:04:27.919Repositorio Institucional Universidad Nacional de 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