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1.
Proc Natl Acad Sci U S A ; 121(2): e2314101120, 2024 Jan 09.
Artículo en Inglés | MEDLINE | ID: mdl-38165935

RESUMEN

Mycobacterium abscessus (Mab), a nontuberculous mycobacterial (NTM) species, is an emerging pathogen with high intrinsic drug resistance. Current standard-of-care therapy results in poor outcomes, demonstrating the urgent need to develop effective antimycobacterial regimens. Through synthetic modification of spectinomycin (SPC), we have identified a distinct structural subclass of N-ethylene linked aminomethyl SPCs (eAmSPCs) that are up to 64-fold more potent against Mab over the parent SPC. Mechanism of action and crystallography studies demonstrate that the eAmSPCs display a mode of ribosomal inhibition consistent with SPC. However, they exert their increased antimicrobial activity through enhanced accumulation, largely by circumventing efflux mechanisms. The N-ethylene linkage within this series plays a critical role in avoiding TetV-mediated efflux, as lead eAmSPC 2593 displays a mere fourfold susceptibility improvement against Mab ΔtetV, in contrast to the 64-fold increase for SPC. Even a minor shortening of the linkage by a single carbon, akin to 1st generation AmSPC 1950, results in a substantial increase in MICs and a 16-fold rise in susceptibility against Mab ΔtetV. These shifts suggest that longer linkages might modify the kinetics of drug expulsion by TetV, ultimately shifting the equilibrium towards heightened intracellular concentrations and enhanced antimicrobial efficacy. Furthermore, lead eAmSPCs were also shown to synergize with various classes of anti-Mab antibiotics and retain activity against clinical isolates and other mycobacterial strains. Encouraging pharmacokinetic profiles coupled with robust efficacy in Mab murine infection models suggest that eAmSPCs hold the potential to be developed into treatments for Mab and other NTM infections.


Asunto(s)
Antiinfecciosos , Infecciones por Mycobacterium no Tuberculosas , Mycobacterium abscessus , Humanos , Animales , Ratones , Espectinomicina/farmacología , Infecciones por Mycobacterium no Tuberculosas/tratamiento farmacológico , Infecciones por Mycobacterium no Tuberculosas/microbiología , Antibacterianos/farmacología , Micobacterias no Tuberculosas , Antiinfecciosos/farmacología , Etilenos/farmacología , Pruebas de Sensibilidad Microbiana
2.
Bioorg Med Chem Lett ; 28(18): 3105-3112, 2018 10 01.
Artículo en Inglés | MEDLINE | ID: mdl-30097368

RESUMEN

ß-Lactams are the most important class of antibiotics, for which the emergence of resistance threatens their utility. As such, we explored the extent to which the tetramic acid motif, frequently found in naturally occurring antibiotics, can be used to generate novel ß-lactam antibiotics with improved antibacterial activity. We synthesized new ampicillin - tetramic acid, cephalosporin - tetramic acid, and cephamycin - tetramic acid analogs and evaluated their activities against problematic Gram-positive and Gram-negative pathogens. Amongst the analogs, a 7-aminocephalosporanic acid analog, 3397, and a 7-amino-3-vinyl cephalosporanic acid, 3436, showed potent activities against S. aureus NRS 70 (MRSA) with MICs of 6.25 µg/mL and 3.13 µg/mL respectively. These new analogs were ≥16-fold more potent than cefaclor and cephalexin. Additionally, a Δ2 cephamycin - tetramic acid analog 3474 which contained a basic guanidinium substituent at the 5-position of the tetramic acid core displayed potent activity against several clinical strains of K. pneumoniae and E. coli.


Asunto(s)
Antibacterianos/farmacología , Escherichia coli/efectos de los fármacos , Klebsiella pneumoniae/efectos de los fármacos , Lactamas/farmacología , Pirrolidinonas/farmacología , Staphylococcus aureus/efectos de los fármacos , Antibacterianos/síntesis química , Antibacterianos/química , Relación Dosis-Respuesta a Droga , Lactamas/química , Pruebas de Sensibilidad Microbiana , Estructura Molecular , Pirrolidinonas/química , Relación Estructura-Actividad
3.
ACS Infect Dis ; 7(12): 3210-3223, 2021 12 10.
Artículo en Inglés | MEDLINE | ID: mdl-34786940

RESUMEN

Fungal fatty acid (FA) synthase and desaturase enzymes are essential for the growth and virulence of human fungal pathogens. These enzymes are structurally distinct from their mammalian counterparts, making them attractive targets for antifungal development. However, there has been little progress in identifying chemotypes that target fungal FA biosynthesis. To accomplish this, we applied a whole-cell-based method known as Target Abundance-based FItness Screening using Candida albicans. Strains with varying levels of FA synthase or desaturase expression were grown in competition to screen a custom small-molecule library. Hit compounds were defined as preferentially inhibiting the growth of the low target-expressing strains. Dose-response experiments confirmed that 16 hits (11 with an acyl hydrazide core) differentially inhibited the growth of strains with an altered desaturase expression, indicating a specific chemical-target interaction. Exogenous unsaturated FAs restored C. albicans growth in the presence of inhibitory concentrations of the most potent acyl hydrazides, further supporting the primary mechanism being inhibition of FA desaturase. A systematic analysis of the structure-activity relationship confirmed the acyl hydrazide core as essential for inhibitory activity. This collection demonstrated broad-spectrum activity against Candida auris and mucormycetes and retained the activity against azole-resistant candida isolates. Finally, a preliminary analysis of toxicity to mammalian cells identified potential lead compounds with desirable selectivities. Collectively, these results establish a scaffold that targets fungal FA biosynthesis with a potential for development into novel therapeutics.


Asunto(s)
Candida auris , Candida , Animales , Antifúngicos/farmacología , Antifúngicos/uso terapéutico , Candida albicans , Ácidos Grasos , Humanos
4.
ACS Infect Dis ; 5(11): 1915-1925, 2019 11 08.
Artículo en Inglés | MEDLINE | ID: mdl-31588734

RESUMEN

Acyldepsipeptides are a unique class of antibiotics that act via allosterically dysregulated activation of the bacterial caseinolytic protease (ClpP). The ability of ClpP activators to kill nongrowing bacteria represents a new opportunity to combat deep-seated biofilm infections. However, the acyldepsipeptide scaffold is subject to rapid metabolism. Herein, we explore alteration of the potentially metabolically reactive α,ß unsaturated acyl chain. Through targeted synthesis, a new class of phenyl urea substituted depsipeptide ClpP activators with improved metabolic stability is described. The ureadepsipeptides are potent activators of Staphylococcus aureus ClpP and show activity against Gram-positive bacteria, including S. aureus biofilms. These studies demonstrate that a phenyl urea motif can successfully mimic the double bond, maintaining potency equivalent to acyldepsipeptides but with decreased metabolic liability. Although removal of the double bond from acyldepsipeptides generally has a significant negative impact on potency, structural studies revealed that the phenyl ureadepsipeptides can retain potency through the formation of a third hydrogen bond between the urea and the key Tyr63 residue in the ClpP activation domain. Ureadepsipeptides represent a new class of ClpP activators with improved drug-like properties, potent antibacterial activity, and the tractability to be further optimized.


Asunto(s)
Antibacterianos/farmacología , Proteínas Bacterianas/metabolismo , Depsipéptidos/química , Endopeptidasa Clp/metabolismo , Activadores de Enzimas/química , Staphylococcus aureus/enzimología , Antibacterianos/química , Antibacterianos/metabolismo , Proteínas Bacterianas/agonistas , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Depsipéptidos/metabolismo , Depsipéptidos/farmacología , Endopeptidasa Clp/química , Endopeptidasa Clp/genética , Activadores de Enzimas/metabolismo , Activadores de Enzimas/farmacología , Dominios Proteicos , Staphylococcus aureus/química , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/genética , Urea/química , Urea/metabolismo
5.
J Antibiot (Tokyo) ; 70(1): 65-72, 2017 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-27189120

RESUMEN

Exploiting iron-uptake pathways by conjugating ß-lactam antibiotics with iron-chelators, such as catechol and hydroxamic acid is a proven strategy to overcome permeability-related resistance in Gram-negative bacteria. As naturally occurring iron-chelating tetramic acids have not been previously examined for this purpose, an exploratory series of novel ampicillin-tetramic acid hybrids that structurally resemble ureidopenicillins was designed and synthesized. The new analogs were evaluated for the ability to chelate iron and their MIC activities determined against a representative panel of clinically significant bacterial pathogens. The tetramic acid ß-lactam hybrids demonstrated a high affinity to iron in the order of 10-30 M3. The hybrids were less active against Gram-positive bacteria. However, against Gram-negative bacteria, their activity was species dependent with several hybrids displaying improved activity over ampicillin against wild-type Pseudomonas aeruginosa. The anti-Gram-negative activities of the hybrids improved in the presence of clavulanic acid revealing that the tetramic acid moiety did not provide added protection against ß-lactamases. In addition, the hybrids were found to be efflux pump substrates as their activities markedly improved against pump-inactivated strains. Unlike the catechol and hydroxamic acid siderophore ß-lactam conjugates, the activities of the hybrids did not improve under iron-deficient conditions. These results suggest that the tetramic acid hybrids gain permeability via different membrane receptors, or they are outcompeted by native bacterial siderophores with stronger affinities for iron. This study provides a foundation for the further exploitation of the tetramic acid moiety to achieve novel ß-lactam anti-Gram-negative agents, providing that efflux and ß-lactamase mediated resistance is addressed.


Asunto(s)
Ampicilina/farmacología , Antibacterianos/farmacología , Bacterias Gramnegativas/efectos de los fármacos , Bacterias Grampositivas/efectos de los fármacos , Pirrolidinonas/farmacología , Ampicilina/administración & dosificación , Ampicilina/síntesis química , Antibacterianos/administración & dosificación , Antibacterianos/síntesis química , Diseño de Fármacos , Farmacorresistencia Bacteriana , Pruebas de Sensibilidad Microbiana , Pirrolidinonas/administración & dosificación , Pirrolidinonas/síntesis química , Sideróforos/metabolismo , Especificidad de la Especie , beta-Lactamasas/metabolismo , beta-Lactamas/administración & dosificación , beta-Lactamas/síntesis química , beta-Lactamas/farmacología
6.
PLoS One ; 10(10): e0139823, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26445268

RESUMEN

We have used RNASeq and qRT-PCR to study mRNA levels for all σ-factors in different Mycobacterium marinum strains under various growth and stress conditions. We also studied their levels in M. marinum from infected fish and mosquito larvae. The annotated σ-factors were expressed and transcripts varied in relation to growth and stress conditions. Some were highly abundant such as sigA, sigB, sigC, sigD, sigE and sigH while others were not. The σ-factor mRNA profiles were similar after heat stress, during infection of fish and mosquito larvae. The similarity also applies to some of the known heat shock genes such as the α-crystallin gene. Therefore, it seems probable that the physiological state of M. marinum is similar when exposed to these different conditions. Moreover, the mosquito larvae data suggest that this is the state that the fish encounter when infected, at least with respect to σ-factor mRNA levels. Comparative genomic analysis of σ-factor gene localizations in three M. marinum strains and Mycobacterium tuberculosis H37Rv revealed chromosomal rearrangements that changed the localization of especially sigA, sigB, sigD, sigE, sigF and sigJ after the divergence of these two species. This may explain the variation in species-specific expression upon exposure to different growth conditions.


Asunto(s)
Proteínas Bacterianas/genética , Respuesta al Choque Térmico/genética , ARN Mensajero/genética , Factor sigma/genética , Estrés Fisiológico/genética , Animales , Culicidae/microbiología , Regulación Bacteriana de la Expresión Génica/genética , Larva/microbiología , Mycobacterium marinum/genética , Mycobacterium tuberculosis/genética , Especificidad de la Especie , Transcripción Genética/genética , alfa-Cristalinas/genética
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