3,5-Dinitrobenzylsulfanyl tetrazoles and 1,3,4-oxadiazoles, previously identified as having high in vitro activities against both replicating and nonreplicating mycobacteria and favorable cytotoxicity and genotoxicity profiles were investigated. First we demonstrated that these compounds act in a deazaflavin-dependent nitroreduction pathway and thus require a nitro group for their activity. Second, we confirmed the necessity of both nitro groups for antimycobacterial activity through extensive structure-activity relationship studies using 32 structural types of analogues, each in a five-membered series. Only the analogues with shifted nitro groups, namely, 2,5-dinitrobenzylsulfanyl oxadiazoles and tetrazoles, maintained high antimycobacterial activity but in this case mainly as a result of DprE1 inhibition. However, these analogues also showed increased toxicity to the mammalian cell line. Thus, both nitro groups in 3,5-dinitrobenzylsulfanyl-containing antimycobacterial agents remain essential for their high efficacy, and further efforts should be directed at finding ways to address the possible toxicity and solubility issues, for example, by targeted delivery.
- MeSH
- antituberkulotika farmakologie chemie MeSH
- mikrobiální testy citlivosti MeSH
- Mycobacterium tuberculosis MeSH
- nitroreduktasy MeSH
- oxadiazoly farmakologie chemie MeSH
- savci MeSH
- tetrazoly farmakologie chemie MeSH
- vztahy mezi strukturou a aktivitou MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Tuberculosis (TB) is a global issue that poses a significant economic burden as a result of the ongoing emergence of drug-resistant strains. The urgent requirement for the development of novel antitubercular drugs can be addressed by targeting specific enzymes. One such enzyme, Mycobacterium tuberculosis (MTB) enoyl-acyl carrier protein (enoyl-ACP) reductase (InhA), plays a crucial role in the survival of the MTB bacterium. In this research study, a series of hybrid compounds combining quinolone and isatin were synthesized and assessed for their effectiveness against MTB, as well as their ability to inhibit the activity of the InhA enzyme in this bacterium. Among the compounds tested, 7a and 5g exhibited the most potent inhibitory activity against MTB, with minimum inhibitory concentration (MIC) values of 55 and 62.5 μg/mL, respectively. These compounds were further evaluated for their inhibitory effects on InhA and demonstrated significant activity compared to the reference drug Isoniazid (INH), with IC50 values of 0.35 ± 0.01 and 1.56 ± 0.06 μM, respectively. Molecular docking studies investigated the interactions between compounds 7a and 5g and the target enzyme, revealing hydrophobic contacts with important amino acid residues in the active site. To further confirm the stability of the complexes formed by 5g and 7a with the target enzyme, molecular dynamic simulations were employed, which demonstrated that both compounds 7a and 5g undergo minor structural changes and remain nearly stable throughout the simulated process, as assessed through RMSD, RMSF, and Rg values.
- MeSH
- antituberkulotika farmakologie chemie MeSH
- bakteriální proteiny metabolismus MeSH
- chinoliny farmakologie MeSH
- isatin farmakologie MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- Mycobacterium tuberculosis MeSH
- oxidoreduktasy metabolismus MeSH
- protein přenášející acyl farmakologie MeSH
- simulace molekulového dockingu MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Článek se zabývá aktuální problematikou tuberkulózy, nemoci, která doprovází lidstvo nejméně 40 tisíciletí. Koncem 20. století ftizeologové, experti na tuberkulózu, optimisticky doufali, že bude do konce druhého tisíciletí vymýcena. Přetrvávající nedobré sociální podmínky v chudých zemích tzv. třetího světa s nedostatečnou kalmetizací, špatným screeningem, a také nástup HIV onemocnění, které výrazně oslabuje imunitu, zvrátily situaci tak, že i dnes, ve 21. století, musíme bojovat s touto infekční nemocí, když navíc čelíme nové výzvě, kmenům mykobakterií tuberkulózy, které jsou odolné vůči běžným antituberkulotikám. V neposlední řadě je nutno zmínit, že do popředí se dostává problém migrace.
The article deals with the current issue of tuberculosis, a disease that has accompanied mankind for at least 40 millennia. In the late 20th century, phthisiologists, experts on tuberculosis, optimistically hoped that it would be eradicated by the end of the second millennium. The persistence of poor social conditions in the poor countries of the so-called Third World, with inadequate calmetisation, poor screening, and the emergence of HIV disease, which significantly weakens immunity, have reversed the situation so that even today, in the 21st century, we have to fight this infectious disease, when, moreover, we are faced with a new challenge, strains of mycobacterium tuberculosis that are resistant to conventional anti-tuberculosis drugs. Last but not least, the problem of migration is coming to the fore.
- MeSH
- antituberkulotika aplikace a dávkování farmakologie terapeutické užití MeSH
- lidé MeSH
- tuberkulinový test MeSH
- tuberkulóza diagnóza epidemiologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
- MeSH
- antituberkulotika farmakologie terapeutické užití MeSH
- atypické mykobakteriální infekce diagnóza terapie MeSH
- BCG vakcína terapeutické užití MeSH
- diagnostické zobrazování metody MeSH
- epidemiologické monitorování MeSH
- epidemiologové MeSH
- lidé MeSH
- lymfadenitida diagnóza etiologie farmakoterapie MeSH
- mikrobiologické techniky metody MeSH
- mykobakteriózy diagnóza farmakoterapie MeSH
- test pomocí interferonu gama metody MeSH
- tuberkulinový test metody MeSH
- tuberkulóza diagnóza farmakoterapie prevence a kontrola MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
Background: Molecular hybridization and isostery are proven approaches in medicinal chemistry, and as such we used them to design novel compounds that we investigated as potential antimycobacterials to combat drug-resistant strains. Methods & results: Prepared N-alkyl-2-(pyrimidine-5-carbonyl)hydrazine-1-carboxamides were cyclized to N-alkyl-5-(pyrimidin-5-yl)-1,3,4-oxadiazol-2-amines along with their analogues. A total of 48 compounds were tested against Mycobacterium tuberculosis H37Rv, Mycobacterium avium and Mycobacterium kansasii, with oxadiazoles and C8-C12 alkyls being the most effective from a concentration of 2 μM. Multidrug-resistant strains were inhibited at same concentrations as the susceptible strain. For the most potent N-dodecyl-5-(pyrimidin-5-yl)-1,3,4-oxadiazol-2-amine, the mechanism of action related to cell wall biosynthesis was investigated. Conclusion: Pyrimidine-1,3,4-oxadiazole hybrids are unique antimycobacterial agents inhibiting mainly M. tuberculosis strains without cross-resistance to current drugs and are thus promising drug candidates.
- MeSH
- aminy farmakologie MeSH
- antibakteriální látky farmakologie MeSH
- antituberkulotika farmakologie chemie MeSH
- mikrobiální testy citlivosti MeSH
- Mycobacterium tuberculosis MeSH
- oxadiazoly farmakologie chemie MeSH
- pyrimidiny farmakologie MeSH
- vztahy mezi strukturou a aktivitou MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- absces etiologie MeSH
- antituberkulotika farmakologie terapeutické užití MeSH
- diagnostické zobrazování MeSH
- glukokortikoidy farmakologie terapeutické užití MeSH
- kojenec MeSH
- lidé MeSH
- mikrobiologické techniky MeSH
- plicní tuberkulóza diagnóza farmakoterapie MeSH
- tuberkulom diagnóza etiologie MeSH
- tuberkulóza páteře chirurgie farmakoterapie MeSH
- tuberkulóza chirurgie diagnóza farmakoterapie MeSH
- výsledek terapie MeSH
- Check Tag
- kojenec MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
Tuberculosis remains a serious killer among infectious diseases due to its incidence, mortality, and occurrence of resistant mycobacterial strains. The challenge to discover new antimycobacterial agents forced us to prepare a series of N-(1-hydroxy-1,3-dihydrobenzo[c][1,2]oxaborol-6-yl)(hetero)aryl-2-carboxamides 1-19 via the acylation of 6-aminobenzo[c][1,2]oxaborol-1(3H)-ol with various activated (hetero)arylcarboxylic acids. These novel compounds have been tested in vitro against a panel of clinically important fungi and bacteria, including mycobacteria. Some of the compounds inhibited the growth of mycobacteria in the range of micromolar concentrations and retained this activity also against multidrug-resistant clinical isolates. Half the maximal inhibitory concentrations against the HepG2 cell line indicated an acceptable toxicological profile. No growth inhibition of other bacteria and fungi demonstrated selectivity of the compounds against mycobacteria. The structure-activity relationships have been derived and supported with a molecular docking study, which confirmed a selectivity toward the potential target leucyl-tRNA synthetase without an impact on the human enzyme. The presented compounds can become important materials in antimycobacterial research.
- MeSH
- amidy chemie farmakologie MeSH
- aminoacyl-tRNA-synthetasy MeSH
- antibakteriální látky farmakologie MeSH
- antiinfekční látky farmakologie MeSH
- antituberkulotika farmakologie MeSH
- houby MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- Mycobacterium tuberculosis MeSH
- simulace molekulového dockingu MeSH
- vztahy mezi strukturou a aktivitou MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Multidrug-resistant tuberculosis (MDR-TB) poses a significant threat to mankind and as such earned its place on the WHO list of priority pathogens. New antimycobacterials with a mechanism of action different to currently used agents are highly required. This study presents the design, synthesis, and biological evaluation of 3-acylaminopyrazine-2-carboxamides derived from a previously reported inhibitor of human prolyl-tRNA synthetase. Compounds were evaluated in vitro against various strains of mycobacteria, pathogenic bacteria, and fungi of clinical significance. In general, high activity against mycobacteria was noted, while the antibacterial and antifungal activity was minimal. The most active compounds were 4'-substituted 3-(benzamido)pyrazine-2-carboxamides, exerting MIC (Minimum Inhibitory Concentration) from 1.95 to 31.25 μg/mL. Detailed structure-activity relationships were established and rationalized in silico with regard to mycobacterial ProRS as a probable target. The active compounds preserved their activity even against multidrug-resistant strains of Mycobacterium tuberculosis. At the same time, they were non-cytotoxic against HepG2 human hepatocellular carcinoma cells. This project is the first step in the successful repurposing of inhibitors of human ProRS to inhibitors of mycobacterial ProRS with antimycobacterial activity.
The emergence and spread of resistant tuberculosis (TB) pose a threat to public health, so it is necessary to diagnose the drug-resistant forms in a clinically short time frame and closely monitor their transmission. In this study, we carried out a first whole genome sequencing (WGS)-based analysis of multidrug resistant (MDR) M. tuberculosis strains to explore the phylogenetic lineages diversity, drug resistance mechanisms, and ongoing transmission chains within the country. In total, 65 isolates phenotypically resistant to at least rifampicin and isoniazid collected in the Czech Republic in 2005-2020 were enrolled for further analysis. The agreement of the results obtained by WGS with phenotypic drug susceptibility testing (pDST) in the determination of resistance to isoniazid, rifampicin, pyrazinamide, streptomycin, second-line injectables and fluoroquinolones was more than 80%. Phylogenetic analysis of WGS data revealed that the majority of MDR M. tuberculosis isolates were the Beijing lineage 2.2.1 (n = 46/65; 70.8%), while the remaining strains belonged to Euro-American lineage. Cluster analysis with a predefined cut-off distance of less than 12 single nucleotide polymorphisms between isolates showed 19 isolates in 6 clusters (clustering rate 29.2%), located mainly in the region of the capital city of Prague. This study highlights the utility of WGS as a high-resolution approach in the diagnosis, characterization of resistance patterns, and molecular-epidemiological analysis of resistant TB in the country.
- MeSH
- antituberkulotika farmakologie terapeutické užití MeSH
- fylogeneze MeSH
- genotyp MeSH
- isoniazid MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- mnohočetná bakteriální léková rezistence genetika MeSH
- multirezistentní tuberkulóza diagnóza farmakoterapie epidemiologie MeSH
- mutace MeSH
- Mycobacterium tuberculosis MeSH
- rifampin MeSH
- sekvenování celého genomu metody MeSH
- tuberkulóza farmakoterapie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
Mycobacterium tuberculosis is an intracellular pathogen and the uptake of the antimycobacterial compounds by host cells is limited. Novel antimycobacterials effective against intracellular bacteria are needed. New N-substituted derivatives of 4-aminosalicylic acid have been designed and evaluated. To achieve intracellular efficacy and selectivity, these compounds were conjugated to tuftsin peptides via oxime or amide bonds. These delivery peptides can target tuftsin- and neuropilin receptor-bearing cells, such as macrophages and various other cells of lung origin. We have demonstrated that the in vitro antimycobacterial activity of the 4-aminosalicylic derivatives against M. tuberculosis H37Rv was preserved in the peptide conjugates. The free drugs were ineffective on infected cells, but the conjugates were active against the intracellular bacteria and have the selectivity on various types of host cells. The intracellular distribution of the carrier peptides was assessed, and the peptides internalize and display mainly in the cytosol in a concentration-dependent manner. The penetration ability of the most promising carrier peptide OT5 was evaluated using Transwell-inserts and spheroids. The pentapeptide exhibited time- and concentration-dependent penetration across the non-contact monolayers. Also, the pentapeptide has a fair penetration rate towards the center of spheroids formed of EBC-1 cells.
- MeSH
- antibakteriální látky farmakologie MeSH
- antituberkulotika chemie farmakologie MeSH
- kyselina aminosalicylová farmakologie MeSH
- mikrobiální testy citlivosti MeSH
- Mycobacterium tuberculosis MeSH
- peptidy chemie MeSH
- pomocné látky farmakologie MeSH
- tuftsin chemie farmakologie MeSH
- Publikační typ
- časopisecké články MeSH
