RT Journal Article
T1 Boldine-Derived Alkaloids Inhibit the Activity of DNA Topoisomerase I and Growth of Mycobacterium tuberculosis
A1 García Esteban, María Teresa
A1 Carreño, David
A1 Tirado Vélez, José Manuel
A1 Ferrándiz, María José
A1 Rodrigues, Liliana
A1 Gracia, Begoña
A1 Amblar, Mónica
A1 Ainsa, José A.
A1 Gonzalez de la Campa, Adela
AB The spread of multidrug-resistant isolates of Mycobacterium tuberculosis requires the discovery of new drugs directed to new targets. In this study, we investigated the activity of two boldine-derived alkaloids, seconeolitsine (SCN) and N-methyl-seconeolitsine (N-SCN), against M. tuberculosis. These compounds have been shown to target DNA topoisomerase I enzyme and inhibit growth of Streptococcus pneumoniae. Both SCN and N-SCN inhibited M. tuberculosis growth at 1.95–15.6 μM, depending on the strain. In M. smegmatis this inhibitory effect correlated with the amount of topoisomerase I in the cell, hence demonstrating that this enzyme is the target for these alkaloids in mycobacteria. The gene coding for topoisomerase I of strain H37Rv (MtbTopoI) was cloned into pQE1 plasmid of Escherichia coli. MtbTopoI was overexpressed with an N-terminal 6-His-tag and purified by affinity chromatography. In vitro inhibition of MtbTopoI activity by SCN and N-SCN was tested using a plasmid relaxation assay. Both SCN and N-SCN inhibited 50% of the enzymatic activity at 5.6 and 8.4 μM, respectively. Cleavage of single-stranded DNA was also inhibited with SCN. The effects on DNA supercoiling were also evaluated in vivo in plasmid-containing cultures of M. tuberculosis. Plasmid supercoiling densities were −0.060 in cells untreated or treated with boldine, and −0.072 in 1 × MIC N-SCN treated cells, respectively, indicating that the plasmid became hypernegatively supercoiled in the presence of N-SCN. Altogether, these results demonstrate that the M. tuberculosis topoisomerase I enzyme is an attractive drug target, and that SCN and N-SCN are promising lead compounds for drug development.
PB Frontiers Media
SN 1664-302X
YR 2018
FD 2018-07-24
LK https://hdl.handle.net/20.500.14352/112403
UL https://hdl.handle.net/20.500.14352/112403
LA eng
NO García MT, Carreño D, Tirado-Vélez JM, Ferrándiz MJ, Rodriguez L, Gracia B, Mónica A, Ainsa JA, De la Campa AG. Boldine-Derived Alkaloids Inhibit the Activity of DNA Topoisomerase I and Growth of Mycobacterium tuberculosis. Front. Microbiol. 2018. 9 (1659)): 1-9.
NO Ministerio de Economía y Competitividad (España)
NO Centro de Investigación Biomédica en Red de Enfemedades Respiratoiras (CIBERES)
NO European Commission
DS Docta Complutense
RD 12 abr 2025