RESUMEN
A proposed treatment for malaria is a combination of fosmidomycin and clindamycin. Both compounds inhibit the methylerythritol 4-phosphate (MEP) pathway, the parasitic source of farnesyl and geranylgeranyl pyrophosphate (FPP and GGPP, respectively). Both FPP and GGPP are crucial for the biosynthesis of several essential metabolites such as ubiquinone and dolichol, as well as for protein prenylation. Dietary prenols, such as farnesol (FOH) and geranylgeraniol (GGOH), can rescue parasites from MEP inhibitors, suggesting the existence of a missing pathway for prenol salvage via phosphorylation. In this study, we identified a gene in the genome of P. falciparum, encoding a transmembrane prenol kinase (PolK) involved in the salvage of FOH and GGOH. The enzyme was expressed in Saccharomyces cerevisiae, and its FOH/GGOH kinase activities were experimentally validated. Furthermore, conditional knockout parasites (Δ-PolK) were created to investigate the biological importance of the FOH/GGOH salvage pathway. Δ-PolK parasites were viable but displayed increased susceptibility to fosmidomycin. Their sensitivity to MEP inhibitors could not be rescued by adding prenols. Additionally, Δ-PolK parasites lost their capability to utilize prenols for protein prenylation. Experiments using culture medium supplemented with whole/delipidated human plasma in transgenic parasites revealed that human plasma has components that can diminish the effectiveness of fosmidomycin. Mass spectrometry tests indicated that both bovine supplements used in culture and human plasma contain GGOH. These findings suggest that the FOH/GGOH salvage pathway might offer an alternate source of isoprenoids for malaria parasites when de novo biosynthesis is inhibited. This study also identifies a novel kind of enzyme related to isoprenoid metabolism.
Asunto(s)
Diterpenos , Fosfomicina/análogos & derivados , Hemiterpenos , Parásitos , Pentanoles , Humanos , Animales , Bovinos , Parásitos/metabolismo , Fosfatos , Terpenos/farmacología , Terpenos/metabolismoRESUMEN
BACKGROUND: Considering the interesting role in the peptidoglycan biosynthesis pathway, the enzyme UDP-N-acetylglucosamine enolpyruvyl transferase is an attractive target to develop new antibacterial agents. It catalyzes the first key step of this pathway and its inhibition leads to bacterial cell death. Fosfomycin is known as the natural inhibitor of MurA. OBJECTIVE: The study aimed to introduce new inhibitors of MurA by virtual screening of different chemical compounds libraries, and test the best scored "virtual hits" against three pathogenic bacteria: Escherichia coli, Bacillus subtilis and Staphylococcus aureus. METHODS: A virtual screening of the structural analogues of fosfomycin downloaded from the Pub- Chem database was performed. Moreover, French National Chemical Library and ZINC database were also utilized to identify new structures different from fosfomycin. FlexX was the software used for this study. The antibacterial testing was divided into two methods: disk diffusion and broth dilution. RESULTS: A set of virtual hits was found to have better energy score than that of fosfomycin, seven of them were tested in vitro. In addition, the disk diffusion method explored four compounds that exhibited antibacterial activity: CID-21680357 (fosfomycin analogue), AB-00005001, ZINC04658565, and ZINC901335. The testing was continued by broth dilution method for both compounds CID-21680357 and ZINC901335 to determine their minimum inhibitory concentrations, and ZINC901335 had the best value with 457µg/ml against Staphylococcus aureus. CONCLUSION: Four compounds were found and proven in silico and in vitro to have antibacterial activity, namely CID-21680357, AB-00005001, ZINC04658565, and ZINC901335.
Asunto(s)
Transferasas Alquil y Aril/antagonistas & inhibidores , Antibacterianos/química , Inhibidores Enzimáticos/química , Fosfomicina/análogos & derivados , Simulación del Acoplamiento Molecular/métodos , Transferasas Alquil y Aril/metabolismo , Antibacterianos/farmacología , Pruebas Antimicrobianas de Difusión por Disco/métodos , Evaluación Preclínica de Medicamentos/métodos , Inhibidores Enzimáticos/farmacología , Escherichia coli/efectos de los fármacos , Escherichia coli/fisiología , Fosfomicina/farmacología , Pruebas de Sensibilidad Microbiana/métodosRESUMEN
A series of eleven double prodrug derivatives of a fosmidomycin surrogate were synthesized and investigated for their ability to inhibit in vitro growth of P. falciparum and M. tuberculosis. A pivaloyloxymethyl (POM) phosphonate prodrug modification was combined with various prodrug derivatisations of the hydroxamate moiety. The majority of compounds showed activity comparable with or inferior to fosmidomycin against P. falciparum. N-benzyl substituted carbamate prodrug 6f was the most active antimalarial analog with an IC50 value of 0.64⯵M. Contrary to fosmidomycin and parent POM-prodrug 5, 2-nitrofuran and 2-nitrothiophene prodrugs 6i and 6j displayed promising antitubercular activities.
Asunto(s)
Antimaláricos/química , Antituberculosos/química , Mycobacterium tuberculosis/efectos de los fármacos , Plasmodium falciparum/efectos de los fármacos , Profármacos/química , Antimaláricos/farmacología , Antituberculosos/farmacología , Carbamatos/química , Evaluación Preclínica de Medicamentos/métodos , Fosfomicina/análogos & derivados , Fosfomicina/farmacología , Humanos , Concentración 50 Inhibidora , Estructura Molecular , Nitrofuranos/química , Profármacos/farmacología , Transducción de Señal , Relación Estructura-ActividadRESUMEN
The continuous development of drug resistance by Plasmodium falciparum, the agent responsible for the most severe forms of malaria, creates the need for the development of novel drugs to fight this disease. Fosmidomycin is an effective antimalarial and potent antibiotic, known to act by inhibiting the enzyme 1-deoxy-d-xylulose-5-phosphate reductoisomerase (DXR), essential for the synthesis of isoprenoids in eubacteria and plasmodia, but not in humans. In this study, novel constrained cyclic prodrug analogues of fosmidomycin were synthesized. One, in which the hydroxamate function is incorporated into a six-membered ring, was found have higher antimalarial activity than fosmidomycin against the chloroquine and mefloquine resistant P. falciparum Dd2 strain. In addition, it showed very low cytotoxicity against cultured human cells.
Asunto(s)
Antimaláricos/síntesis química , Antimaláricos/farmacología , Fosfomicina/análogos & derivados , Profármacos/síntesis química , Profármacos/farmacología , Evaluación Preclínica de Medicamentos , Fosfomicina/síntesis química , Fosfomicina/farmacología , HumanosRESUMEN
The biochemical pathway to hypericin biosynthesis is presumed to be polyketide synthase (PKS) mediated, but it has not been experimentally validated, and no alternate route (chorismate/o-succinylbenzoate pathway) has been analyzed. We report here our earlier developed auxin inducible culture systems of Hypericum hookerianum as a model, to study the metabolic pathway to hypericin synthesis. Inhibitors of the alternate pathway at varying concentrations showed steady synthesis of total hypericins with means of 2.80±0.22, 18.75±0.01; 16.39±3.75, 29.60±1.90 (mevinolin) 2.53±0.10, 18.12±0.56; 0.14±0.01, 14.28±1.11 (fosmidomycin) and 2.7±0.35, 18.75±0.61; 0.14±0.01, 12.80±1.09 mg g(-1) DW (glyphosate) in the control and auxin-induced shoot and shoot-forming callus cultures, respectively. SSH analysis classified the differentially expressed sequences into protein synthesis (38%), modification (20%), electron transport (9%) and remaining as unclassified (11%) and unknown proteins (22%). Functional annotation of sequences indicates the presence of additional protein components besides PKS activity. Our results demonstrate direct biochemical and molecular evidence of PKS hypothesis of hypericin biosynthesis for the first time.
Asunto(s)
Hypericum/metabolismo , Perileno/análogos & derivados , Antracenos , Emodina/química , Emodina/metabolismo , Fosfomicina/administración & dosificación , Fosfomicina/análogos & derivados , Glicina/administración & dosificación , Glicina/análogos & derivados , Hypericum/efectos de los fármacos , Ácidos Indolacéticos/farmacología , Lovastatina/administración & dosificación , Perileno/química , Perileno/aislamiento & purificación , Perileno/metabolismo , Brotes de la Planta/efectos de los fármacos , Brotes de la Planta/crecimiento & desarrollo , Brotes de la Planta/metabolismo , Sintasas Poliquetidas/metabolismo , Técnicas de Cultivo de Tejidos , GlifosatoRESUMEN
The methylerythritol phosphate (MEP) pathway in plants produces the prenyl precursors for all plastidic isoprenoids, including carotenoids and quinones. The MEP pathway is also responsible for synthesis of approximately 600 Tg of isoprene per year, the largest non-methane hydrocarbon flux into the atmosphere. There have been few studies of the regulation of the MEP pathway in plants under physiological conditions. In this study, we combined gas exchange techniques and high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS-MS) and measured the profile of MEP pathway metabolites under different conditions. We report that in the MEP pathway, metabolites immediately preceding steps requiring reducing power were in high concentration. Inhibition of the MEP pathway by fosmidomycin caused deoxyxylulose phosphate accumulation in leaves as expected. Evidence is presented that accumulation of MEP pathway intermediates, primarily methylerythritol cyclodiphosphate, is responsible for the post-illumination isoprene burst phenomenon. Pools of intermediate metabolites stayed at approximately the same level 10 min after light was turned off, but declined eventually under prolonged darkness. In contrast, a strong inhibition of the second-to-last step of the MEP pathway caused suppression of isoprene emission in pure N(2). Our study suggests that reducing equivalents may be a key regulator of the MEP pathway and therefore isoprene emission from leaves.
Asunto(s)
Butadienos/metabolismo , Eritritol/análogos & derivados , Eritritol/metabolismo , Hemiterpenos/metabolismo , Luz , Redes y Vías Metabólicas/efectos de la radiación , Metaboloma , Pentanos/metabolismo , Hojas de la Planta/metabolismo , Populus/metabolismo , Aclimatación/efectos de los fármacos , Aclimatación/efectos de la radiación , Cromatografía Líquida de Alta Presión , Oscuridad , Eritritol/química , Fosfomicina/análogos & derivados , Fosfomicina/farmacología , Espectrometría de Masas , Redes y Vías Metabólicas/efectos de los fármacos , Metaboloma/efectos de los fármacos , Metaboloma/efectos de la radiación , Nitrógeno/farmacología , Extractos Vegetales , Hojas de la Planta/efectos de los fármacos , Hojas de la Planta/efectos de la radiación , Populus/efectos de la radiación , Estándares de Referencia , Factores de TiempoRESUMEN
Fosmidomycin (100 micromol x L(-1)) which is the effective inhibitor of DXR, key enzyme in terpenoid MEP pathway, was used to treat with hairy roots of Salvia miltiorrhiza. The treated roots were harvested at 2, 4, 6, 8, 10, 16 and 21 d, mRNA level of SmDXR and tanshinone content in treated and negative control groups were detected. Results found that, after treated with fosmidomycin, color of S. miltiorrhiza hairy roots grew pale gradually comparing with controls; mRNA level of SmDXR in hairy roots varied as a shape of parabolic and the highest value achieved at the sixth day after treatment, then it decreased gradually; Content of four kinds of tanshinones were detected. Among of the four kinds of tanshinones, Tanshinone I content changed relatively little, while content of dihydrotanshinone I, cryptotanshinone and tanshinone II (A) decreased gradually in 21 days. The content of total tanshinones in NC groups was 5, 63 times more than FOS-treated roots in the 21th day. The previous results showed that SmDXR played an important role in the accumulation of tanshinone content in MEP pathway. Once the mRNA level of SmDXR was suppressed, the accumulation of secondary metabolites will be significantly affected.
Asunto(s)
Abietanos/metabolismo , Fosfomicina/análogos & derivados , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/metabolismo , Salvia miltiorrhiza/efectos de los fármacos , Salvia miltiorrhiza/metabolismo , Isomerasas Aldosa-Cetosa/genética , Fosfomicina/farmacología , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Raíces de Plantas/crecimiento & desarrollo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Salvia miltiorrhiza/genética , Salvia miltiorrhiza/crecimiento & desarrollo , Factores de TiempoAsunto(s)
Farmacorresistencia Bacteriana Múltiple/efectos de los fármacos , Fosfomicina/análogos & derivados , Chryseobacterium/efectos de los fármacos , Chryseobacterium/metabolismo , Enterobacter cloacae/efectos de los fármacos , Enterobacter cloacae/metabolismo , Eritritol/análogos & derivados , Eritritol/metabolismo , Escherichia coli/efectos de los fármacos , Escherichia coli/metabolismo , Fosfomicina/farmacología , Klebsiella pneumoniae/efectos de los fármacos , Klebsiella pneumoniae/metabolismo , Pruebas de Sensibilidad Microbiana , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/metabolismo , Fosfatos de Azúcar/metabolismo , Terpenos/metabolismoRESUMEN
Fosmidomycin, which acts through inhibition of 1-deoxy-D-xylulose phosphate reductoisomerase (DXR) in the non-mevalonate pathway, represents a valuable recent addition to the armamentarium against uncomplicated malaria. In this paper, we describe the synthesis and biological evaluation of E- and Z-alpha,beta-unsaturated alpha-aryl-substituted analogues of FR900098, a fosmidomycin congener, utilizing a Stille or a Suzuki coupling to introduce the aryl group. In contrast with our expectations based on the promising activity earlier observed for several alpha-substituted fosmidomycin analogues, all synthesized analogues exhibited much lower binding affinity for DXR than fosmidomycin.
Asunto(s)
Isomerasas Aldosa-Cetosa/antagonistas & inhibidores , Antimaláricos/síntesis química , Química Farmacéutica/métodos , Inhibidores Enzimáticos/farmacología , Fosfomicina/análogos & derivados , Malaria/tratamiento farmacológico , Complejos Multienzimáticos/antagonistas & inhibidores , Oxidorreductasas/antagonistas & inhibidores , Animales , Antimaláricos/farmacología , Diseño de Fármacos , Evaluación Preclínica de Medicamentos , Fosfomicina/síntesis química , Fosfomicina/química , Concentración 50 Inhibidora , Modelos Químicos , Estructura Molecular , Plasmodium falciparum/enzimología , Relación Estructura-ActividadRESUMEN
Fosmidomycin is effective against malaria, but it needs to be given for > or =4 days when used alone. We conducted a study of 50 children with Plasmodium falciparum malaria to evaluate the safety and efficacy of consecutively shortened regimens of artesunate-fosmidomycin (1 to 2 mg/kg of body weight and 30 mg/kg of body weight, respectively; doses given every 12 hours). All dosing regimens were well tolerated. Artesunate-fosmidomycin acted rapidly, resulting in consolidated geometric mean parasite and fever clearance times of 24 h and 15 h, respectively. Treatment regimens of > or =2 days led to cure ratios of 100% by day 14 (39/39; 95% confidence interval [95% CI], 91% to 100%). Most importantly, the 3-day regimen achieved 100% cure on day 28 (10/10; 95% CI, 69% to 100%). Treatment with artesunate-fosmidomycin was associated with transient grade I or II neutropenia (absolute neutrophil counts of 750 to 1,200/microl and 400 to 749/microl, respectively) in six or two patients, respectively. Artesunate-fosmidomycin demonstrates the feasibility and potential value of short-course artemisinin-based combination chemotherapy with rapidly eliminated combination partners.
Asunto(s)
Antimaláricos/uso terapéutico , Artemisininas/uso terapéutico , Fosfomicina/análogos & derivados , Malaria Falciparum/tratamiento farmacológico , Sesquiterpenos/uso terapéutico , Antimaláricos/efectos adversos , Artemisininas/efectos adversos , Artesunato , Química Farmacéutica , Niño , Estudios de Cohortes , Relación Dosis-Respuesta a Droga , Combinación de Medicamentos , Determinación de Punto Final , Femenino , Fosfomicina/efectos adversos , Fosfomicina/uso terapéutico , Humanos , Malaria Falciparum/sangre , Malaria Falciparum/parasitología , Masculino , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Sesquiterpenos/efectos adversosRESUMEN
The aim of our study was to verify if the empiric therapy with a single dose of 3 g fosfomycin tromethamine in patients with uncomplicated urinary tract infections (UTIs) was able to clinically resolveand to microbiologically eradicate the infection. A total of 387 out of the 400 patients (274 cases with acute and 113 cases with recurrent uncomplicated UTIs) were enrolled in the clinical study. Clinical and microbiological assessments were performed before and at 8-10 days after the administration. At follow-up high clinical recovery (88.9%) and bacteriological (94.9%) eradication rates were achieved. Gastrointestinal side effects were found in only 4.3% of patients. In conclusion, a single-dose administration regimen of fosfomycin tromethamine should be encouraged as a first choice of drug therapy for uncomplicated UTIs.
Asunto(s)
Antiinfecciosos Urinarios/uso terapéutico , Fosfomicina/análogos & derivados , Trometamina/análogos & derivados , Infecciones Urinarias/tratamiento farmacológico , Adolescente , Adulto , Antiinfecciosos Urinarios/administración & dosificación , Esquema de Medicación , Femenino , Fosfomicina/administración & dosificación , Fosfomicina/uso terapéutico , Humanos , Masculino , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Trometamina/administración & dosificación , Trometamina/uso terapéuticoRESUMEN
BACKGROUND: Isopentenyl diphosphate (IPP), a common biosynthetic precursor to the labdane diterpene forskolin, has been biosynthesised via a non-mevalonate pathway. Geranylgeranyl diphosphate (GGPP) synthase is an important branch point enzyme in terpenoid biosynthesis. Therefore, GGPP synthase is thought to be a key enzyme in biosynthesis of forskolin. Herein we report the first confirmation of the GGPP synthase gene in Coleus forskohlii Briq. RESULTS: The open reading frame for full-length GGPP synthase encodes a protein of 359 amino acids, in which 1,077 nucleotides long with calculated molecular mass of 39.3 kDa. Alignments of C. forskohlii GGPP synthase amino acid sequences revealed high homologies with other plant GGPP synthases. Several highly conserved regions, including two aspartate-rich motifs were identified. Transient expression of the N-terminal region of C. forskohlii GGPP synthase-GFP fusion protein in tobacco cells demonstrated subcellular localization in the chloroplast. Carotenoid production was observed in Escherichia coli harboring pACCAR25DeltacrtE from Erwinia uredovora and plasmid carrying C. forskohlii GGPP synthase. These results suggested that cDNA encoded functional GGPP synthase. Furthermore, C. forskohlii GGPP synthase expression was strong in leaves, decreased in stems and very little expression was observed in roots. CONCLUSION: This investigation proposed that forskolin was synthesised via a non-mevalonate pathway. GGPP synthase is thought to be involved in the biosynthesis of forskolin, which is primarily synthesised in the leaves and subsequently accumulates in the stems and roots.
Asunto(s)
Transferasas Alquil y Aril/genética , Coleus/genética , Fosfomicina/análogos & derivados , Transferasas Alquil y Aril/metabolismo , Secuencia de Aminoácidos , Línea Celular , Clonación Molecular , Coleus/enzimología , Coleus/metabolismo , Colforsina/metabolismo , ADN Complementario/química , ADN Complementario/genética , Electroforesis en Gel de Poliacrilamida , Escherichia coli/genética , Fosfomicina/farmacología , Eliminación de Gen , Regulación Enzimológica de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Prueba de Complementación Genética , Geranilgeranil-Difosfato Geranilgeraniltransferasa , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Microscopía Fluorescente , Datos de Secuencia Molecular , Hojas de la Planta/genética , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/genética , Raíces de Plantas/metabolismo , Tallos de la Planta/genética , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Alineación de Secuencia , Análisis de Secuencia de ADN , Homología de Secuencia de Aminoácido , Nicotiana/citología , TransfecciónRESUMEN
Approximately 40% of the world population live in areas with the risk of malaria. Each year, 300-500 million people suffer from acute malaria, and 0.5-2.5 million die from the disease. Although malaria has been widely eradicated in many parts of the world, the global number of cases continues to rise. The most important reason for this alarming situation is the rapid spread of malaria parasites that are resistant to antimalarial drugs, especially chloroquine, which is by far the most frequently used. The development of new antimalarial drugs has been neglected since the 1970s owing to the end colonialism, changes in the areas of military engagement, and the restricted market potential. Only in recent years, in part supported by public funding programs, has interest in the development of antimalarial drugs been renewed. New data available from the recently sequenced genome of the malaria parasite Plasmodium falciparum and the application of methods of modern drug design promise to bring significant development in the fight against this disease.
Asunto(s)
Antimaláricos/farmacología , Antimaláricos/uso terapéutico , Fosfomicina/análogos & derivados , Proguanil/análogos & derivados , Aminoquinolinas/farmacología , Aminoquinolinas/uso terapéutico , Animales , Artemisininas/farmacología , Artemisininas/uso terapéutico , Artesunato , Atovacuona , Ensayos Clínicos como Asunto , Dapsona/farmacología , Dapsona/uso terapéutico , Combinación de Medicamentos , Diseño de Fármacos , Resistencia a Medicamentos/genética , Etanolaminas/farmacología , Etanolaminas/orina , Flavonoides/farmacología , Flavonoides/uso terapéutico , Fluorenos/farmacología , Fluorenos/orina , Fosfomicina/farmacología , Fosfomicina/uso terapéutico , Salud Global , Humanos , Lumefantrina , Malaria/tratamiento farmacológico , Malaria/epidemiología , Malaria/prevención & control , Naftoquinonas/farmacología , Naftoquinonas/uso terapéutico , Naftiridinas/farmacología , Naftiridinas/uso terapéutico , Plasmodium falciparum/efectos de los fármacos , Plasmodium falciparum/genética , Plasmodium falciparum/fisiología , Proguanil/farmacología , Proguanil/uso terapéutico , Sesquiterpenos/farmacología , Sesquiterpenos/uso terapéuticoRESUMEN
In malaria parasites, isoprenoids are synthesised by the mevalonate independent 1-deoxy- D-xylulose 5-phosphate (DOXP) pathway. Fosmidomycin, a natural antibiotic originally developed for the treatment of bacterial infections, represents an inhibitor of DOXP reductoisomerase, an essential enzyme of this pathway. In recent clinical studies it was shown that fosmidomycin is effective in curing uncomplicated Plasmodium falciparum malaria in humans. The treatment was well tolerated and resulted in a fast parasite and fever clearance. However, the high rate of recrudescence precludes the use of fosmidomycin as a monotherapy. In drug combination studies, synergy of fosmidomycin with clindamycin was observed. Clinical studies with a fosmidomycin-clindamycin combination are currently ongoing.
Asunto(s)
Antimaláricos/uso terapéutico , Fosfomicina/análogos & derivados , Fosfomicina/uso terapéutico , Malaria Falciparum/tratamiento farmacológico , Animales , Antimaláricos/farmacocinética , Antimaláricos/farmacología , Química Farmacéutica , Clindamicina/farmacología , Clindamicina/uso terapéutico , Ensayos Clínicos como Asunto , Evaluación Preclínica de Medicamentos , Sinergismo Farmacológico , Quimioterapia Combinada , Fosfomicina/farmacocinética , Fosfomicina/farmacología , Humanos , Pentosafosfatos/metabolismo , Plasmodium/efectos de los fármacos , Plasmodium falciparum/efectos de los fármacos , Plasmodium falciparum/metabolismo , Terpenos/metabolismoRESUMEN
The recently discovered 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway for the biosynthesis of plastid isoprenoids (including carotenoids) is not fully elucidated yet despite its central importance for plant life. It is known, however, that the first reaction completely specific to the pathway is the conversion of 1-deoxy-D-xylulose 5-phosphate (DXP) into MEP by the enzyme DXP reductoisomerase (DXR). We have identified a tomato cDNA encoding a protein with homology to DXR and in vivo activity, and show that the levels of the corresponding DXR mRNA and encoded protein in fruit tissues are similar before and during the massive accumulation of carotenoids characteristic of fruit ripening. The results are consistent with a non-limiting role of DXR, and support previous work proposing DXP synthase (DXS) as the first regulatory enzyme for plastid isoprenoid biosynthesis in tomato fruit. Inhibition of DXR activity by fosmidomycin showed that plastid isoprenoid biosynthesis is required for tomato fruit carotenogenesis but not for other ripening processes. In addition, dormancy was reduced in seeds from fosmidomycin-treated fruit but not in seeds from the tomato yellow ripe mutant (defective in phytoene synthase-1, PSY1), suggesting that the isoform PSY2 might channel the production of carotenoids for abscisic acid biosynthesis. Furthermore, the complete arrest of tomato seedling development using fosmidomycin confirms a key role of the MEP pathway in plant development.
Asunto(s)
Isomerasas Aldosa-Cetosa/biosíntesis , Fosfomicina/análogos & derivados , Complejos Multienzimáticos/biosíntesis , Oxidorreductasas/biosíntesis , Plastidios/metabolismo , Solanum lycopersicum/fisiología , Terpenos/metabolismo , Isomerasas Aldosa-Cetosa/química , Isomerasas Aldosa-Cetosa/genética , Secuencia de Aminoácidos , Secuencia de Bases , Cartilla de ADN , ADN Complementario , Fosfomicina/farmacología , Hibridación in Situ , Solanum lycopersicum/metabolismo , Datos de Secuencia Molecular , Complejos Multienzimáticos/química , Complejos Multienzimáticos/genética , Oxidorreductasas/química , Oxidorreductasas/genética , ARN Mensajero/genética , Homología de Secuencia de AminoácidoRESUMEN
As the antibiotic arsenal expands with the development of new oral agents, selecting appropriate therapy is becoming more difficult. What are the therapeutic indications for the newer antimicrobials, and what advantages do they have over traditional agents? Drs Gleckman and De La Rosa discuss current issues related to antibiotic treatment and offer specific recommendations for the best use of the newer agents.
Asunto(s)
Antiinfecciosos/uso terapéutico , Infecciones/tratamiento farmacológico , Administración Oral , Antibacterianos/uso terapéutico , Antiinfecciosos/administración & dosificación , Cefalosporinas/uso terapéutico , Fluoroquinolonas , Fosfomicina/análogos & derivados , Fosfomicina/uso terapéutico , Humanos , Macrólidos , Cooperación del Paciente , Trometamina/análogos & derivados , Trometamina/uso terapéuticoRESUMEN
Fosfomycin tromethamine is an orally administered fosfomycin that may be used for single-dose therapy of uncomplicated urinary tract infections. At breakpoint concentrations [< or = 128 micrograms/ml plus 25 micrograms/ml glucose-6-phosphate (G-6-P)], fosfomycin tromethamine inhibited > 90% of the 350 bacterial isolates tested. When testing Escherichia coli, Klebsiella spp., and Enterobacter spp., we note that the performance of fosfomycin disks improved when G-6-P was added to the disks. The interpretive error rates were minimized when 200-micrograms fosfomycin disks were supplemented with either 50 or 100 micrograms G-6-P. Using < or = 128 and > or = 256 micrograms/ml as the susceptible and resistant MIC breakpoints, respectively, the regression-analysis-derived disk diffusion zone diameter breakpoints for the 200-micrograms fosfomycin disk supplemented with 50 micrograms of G-6-P are as follows: susceptible, > or = 16 mm; intermediate, 13-15 mm; and resistant, < or = 12 mm.
Asunto(s)
Bacterias/efectos de los fármacos , Fosfomicina/análogos & derivados , Pruebas de Sensibilidad Microbiana , Trometamina/análogos & derivados , Estudios de Evaluación como Asunto , Fosfomicina/farmacología , Glucosa-6-Fosfato , Glucofosfatos , Análisis de Regresión , Trometamina/farmacología , Sistema Urinario/microbiologíaRESUMEN
The response to fosmidomycin of four strains of Escherichia coli was studied in an in vitro model of the treatment of bacterial cystitis. Three susceptible strains of E. coli responded well to relatively low concentrations of fosmidomycin: doses achieving peak concentrations of 50 or 250 mg/l suppressed bacterial growth for 13 h or more; however, when the surviving bacteria were challenged with a second dose, a reduced response was observed. When a fully resistant strain was exposed to fosmidomycin, bacterial growth was also suppressed for 13 h or more, even when the peak concentration achieved was below the conventionally determined minimum inhibitory concentration. Resistant variants which emerged after exposure to fosmidomycin were also resistant to fosfomycin in the absence of the potentiating agent, glucose-6-phosphate. In the presence of glucose-6-phosphate, complete (or partial) susceptibility to fosmidomycin and fosfomycin was retained by three of the four strains. These results suggest that fosmidomycin and fosfomycin are transported into E. coli by a similar mechanism, and that deletion of the hexose phosphate transport system does not occur following exposure to fosmidomycin in the absence of glucose-6-phosphate.