Revisión por pares / joruiz@clinic.ub.es / Furazolidone-resistant mutants were obtained from four clinical isolates of diarrhoeagenic Escherichia coli. The stability of the resistance and the frequency of mutation were established. The minimal inhibitory concentration of furazolidone, nitrofurantoin, nalidixic acid, ampicillin, chloramphenicol and tetracycline was established both in the presence and absence of the efflux pump inhibitor Phe-Arg-β-Naphtylamyde. The presence of mutations in the nitroreductase genes nfsA and nfsB was analysed by PCR; sequencing and their enzymatic activity was assessed by a spectrophotometric assay. Alterations in outer membrane proteins were studied by SDS-PAGE. The frequency of mutation ranged from <9.6 × 10-10 to 9.59 × 10-7 . Neither an effect on efflux pumps inhibited by Phe-Arg-β-Naphtylamyde nor cross-resistance with the antibiotics studied was observed. Nineteen mutants (52.94%) presented mutations in the nitroreductase-encoding genes: 17 in the nfsA gene (15 mutants with an internal stop codon, 2 with amino acid changes), 2 in the nfsB (all amino acid changes). Alterations in the outer membrane proteins OmpA and OmpW were also observed. Although more studies are necessary to find other resistance mechanisms, present data showed the low potential of selecting furazolidone-resistant mutants, together with the lack of cross-resistance with unrelated antimicrobial agents.

Food additives, drugs and growth-enhancing compounds are prominent tools in the production of sufficient quantities of affordable food. Thus, food safety has become a main concern of many countries for protection of their population's health. Analytical chemical measurements are increasingly important to ensure consumer protection, particularly in the field of antibiotic residues. International comparability and reliability of measurement results can be achieved by establishing and demonstrating the metrological traceability of those results to the International System of Units (SI). To ensure the metrological traceability of measurement results, reference methods and certified reference materials (CRMs) are needed. Nitrofuran antibiotic drugs used for the treatment of bacterial and protozoan infections in animals were studied in this thesis. A high accuracy reference method and an appropriate CRM for the detection of nitrofurans in prawns have been developed. A reference method for the measurement of mass fractions of nitrofuran metabolites 3-amino-5-methyl-morpholino-2-oxazolidinone (AMOZ), 3-amino-2-oxazolidinone (AOZ), semicarbazide (SEM) and 1-aminohydantoin (AHD) has been developed utilising an exact matching double isotope dilution mass spectrometry (IDMS) method. A feasibility study on a fortified reference material was carried out by preparing fortified samples containing the four nitrofuran metabolites. The stability testing results showed that the analytes in the freeze-dried matrix were more stable than in the wet form. Freeze-dried certified reference materials of nitrofurans in prawns have been produced; an incurred AOZ (CRM_P1) and incurred AOZ and fortified SEM (CRM_P2). The reference method developed was applied for the characterisation of certified reference materials for homogeneity, stability study and certification. The prepared certified reference materials were found to be homogeneous and remained stable under normal transport conditions. The measurement uncertainty of the measurement results obtained by the reference method developed was determined by thoroughly examining all possible sources of potential bias and precision effects. An initial measurement uncertainty for certified values of AOZ in both materials has also been estimated. Metrological traceability of measurement results obtained by the developed reference method and the reference value of the prepared certified reference materials has been established through the use of the traceable primary ratio method of exact matching double IDMS, the use of certified nitrofuran metabolite standards, and gravimetric preparation of samples.

The popularity of capillary electrophoresis (CE) as a separation technique has been established over the years. CE offers the advantages of high resolution, high separation efficiency, fast approaches of method development, a range of operational modes and low consumption of reagents. The strategy employed here for the development of chromatographic separations involved the utilization of experimental designs, multi-linear regression and response surface methodology to build empirical models that related the chromatographic quality to the factors influencing the separation. Separation of Nitrofuran antibiotics (NFAs) and their metabolites (NFMs) by using micellar electrokinetic capillary chromatography was successfully completed. The best conditions found to give optimum resolution from the optimization study was pH 9.0, 80 mM SDC concentration, 16 kV with running buffer consisting of 20 mM borate and 20 mM phosphate concentration using a 73 cm x 75 ƒÝm column, resulting in completely resolved NFAs and NFMs within 16 min. It is interesting that all the compounds can be reliably separated with the one mixture, and single CE condition. Whilst all antibiotics have shorter migration time than their respective derivatised metabolites, as a group apart from nitrofurantoin the antibiotics elute before the metabolites. The analytical figures of merit for CE analysis exhibited excellent reproducibility of absolute and relative migration times, and acceptable reproducibility of relative response areas. Successful separation of metabolite derivatives was achieved when the developed method was applied to a spiked prawn sample. The chiral separation of Triadimenol was successfully completed using micellar electrokinetic capillary chromatography. The best conditions found to give optimum resolution from the optimisation study were pH 6.0, 20% methanol, 50 mM SDS concentration, 18 kV with running buffer consisting of 20 mM borate and 20 mM phosphate concentration using a 64.5 cm x 50 ƒÝm column, resulted in baseline resolution of all Triadimenol isomers within 18 min. The optimised separation conditions were applied to a blank grape sample and to a spiked grape sample. No peaks were observed in the blank grape sample whereas the spiked grape sample had two diastereoismer peaks with poor detection sensitivity. Increase in detection sensitivity is necessary to determine the possibility of resolution of all the isomers of Triadimenol, in the spiked grape sample and the blank. Online preconcentration techniques were attempted to for Triadimenol isomer separation. When using online preconcentration technique of sweeping, a 30-fold increase in detection sensitivity of Triadimenol was observed compared to MEKC mode. However enantiomer separation was not possible with sulfated-£]-CD chiral selector. The best conditions were found to be pH 2.5, 50 mM SDS concentration, -20 kV with running buffer consisting of 20 mM phosphate concentration, using a 64.5 cm x 50 ƒÝm column, resulting in diastereoisomer separation within 8 min. Final stage of the project was to create stationary phase beds in capillaries and micro-channels that could be removed and re-created, thus providing a fresh stationary phase. The replaceable stationary phase (RSP) can be used as an operating mode of CE/CEC. Preparation of reversible stationary phase (RSP) inside the capillary column was successfully performed using low methoxy pectin (LMP). LMP renders a capability of reversible thermogelation. Electroosmotic flow (EOF) and sufficient hydrophobicity of LMP gel allow separation of analyates. The porosity of LMP RSP was adequate to support EOF. Successful separation with good reproducibility of areas and migration times was obtained for Caffeine, Aspartame, Benzoic acid, Saccharine (CABS) mixture and NFAs. After performing continuous analyses, the aging of RSP was observed. Temperature was the ¡¥switch¡¦, which applied to remove aged RSP. RSP was recreated for further analysis of analytes. RSP was UV transparent, capable of handling various analytes and diff erent buffer electrolytes including aqueous-organic solvents.

Part I: Target identification studies are one of the most difficult but rewarding challenges in chemical biology. Part I of this thesis describes target identification studies for 5-nitrofuran containing hits. The 5-nitrofurans used in this study were identified in a phenotypic screen for compounds that induced melanocyte cells death in zebrafish. Chapter 1 provides brief overviews on three related areas of the project: 1) the use of zebrafish as a model organism in drug discovery; 2) phenotypic screening using zebrafish and 3) the strategies used in target identification studies. Chapter 2 describes the synthesis of and SAR studies on two series of 5-nitrofuran containing analogues. The design and preparation of biotinylated chemical probes based on the SAR data is also described. These chemical tools are then used in affinity chromatography studies and genetic validation of a potential target (zebrafish Aldh2) of the 5-nitrofuran compounds is reported. Chapter 3 provides a review of the biological and chemical processes that human ALDHs are known to mediate. In addition, small molecules that modulate ALDH2 activity are reviewed. A detailed study of the interaction between 5-nitrofurans and human ALDH2 including in vitro enzymatic assays is described leading to the conclusion that the 5- nitrofurans under study are substrates of human ALDH2. Further mechanism of action investigations using model reactions are also presented. Chapter 4 introduces the use of 5-nitrofuran containing drugs in the clinic and highlights the reported side-effects. Further investigation of the interaction between ALDH2 and 5- nitrofurans in zebrafish and yeast using ALDH2 inhibitors is described. Based on these results, a combination therapy strategy is proposed. Finally, the trypanocidal activity of the newly synthesised 5-nitrofurans is discussed. Experimental details and future work for Part I are presented in Chapters 5 and 6 respectively. Part II: Human sirtuins are associated with various biological functions and diseases, including cancer and neurodegeneration. Previous work from the Westwood Lab has led to the discovery of the tenovins that act as inhibitors of SIRT1 and SIRT2. Part II of the thesis reports the development of potent fixed ring tenovin analogues with high SIRT2 selectivity. Chapter 7 provides a brief review of the biology of human SIRT2 and the reported SIRT2 inhibitors available to date. This is followed by a short summary of the previous work on the tenovins in the Westwood Lab and the design of the fixed ring tenovin analogues. Chapter 8 describes the synthesis of three series of fixed ring tenovin analogues. SAR data is generated based on in vitro enzymatic assays against both SIRT1 and SIRT2 and the prepared analogues showed relatively high potency and selectivity against SIRT2. Further cell-based deacetylation assay are also discussed. All the experimental details are reported in Chapter 9 and Chapter 10 provides with conclusions and proposed future work.

Nitrofurans have been widely used either in waterbath or feed additives for the prevention and treatment of aquatic products. The European Union was able to assign a maximum residue limit and prohibited nitrofurans used to animals in 1995, because of the potential carcinogenic effects of their residues on human health. This study is focusing on the analytical method of four kinds of commonly used nitrofurans and corresponding residual metabolites by LC-MS/MS. The detection limits of furazolidone, furaltadone, nitrofurazone and nitrofurntoin were 6.11, 3.63, 4.52 and 6.20 £gg kg-1,respectively. The detection limits of AOZ, AMOZ, SC and AH were 0.23, 0.30, 0.36, 0.53 £gg kg-1, respectively. The lightness is the main factor to cause the decomposition of nitrofurans. It is not significant for temperature to depredate nitrofurans. The adsorbtion of metabolites by the plastic tube was in the extraction procedure. Equipments in glass are suggested to be used for the sample pretreatment and plastic meterials are averted to be exercised. About the comparation of determination of AOZ by ELISA and LC-MS/MS. The result demonstrated that the ELISA method might overestimate the residual AOZ content at low concentrations. The detection limit and recovery of the known addition were 0.05 £gg kg-1 and 108% for the LC-MS/MS method and 0.31 £gg kg-1 and 305% for the ELISA method, respectively. The amounts of residual nitrofurans and metabolites in muscle, liver, gill and skin tissue of tilapia which were treated in different conditions were compared. The depletion data of bathing treatment group obtained showed similar be haviors of furazolidone, furaltadone, nitrofurazone, nitrofurantoin in tilapia which the residual time was less than 24 hr. The amounts of residual nitrofurans appeared the highest concentration in gill and the lowest concentration in muscle. Bonded residues of metabolites can be detected for at least 4 weeks after administration in muscle, skin, liver and gill. The concentrations of residual bonded metabolites were higher than non-bonded metabolites in gill and muscle besides liver during depletion periods. After bathing medication, there were more residual nitrofurans and corresponding metabolites in sea water tilapia than fresh water group, because sea water fish survives in high osmotic condition to reduce their urination. Nitrofurans and metabolites were deconstructed by enzyme in gills, livers, intestines and muscles. Then tissues of fish accumulated nitrofurans and metabolites soon after medication. The maturity of fish is one of facters to effect different residual concentration during depletion periods. Liver is the main tissue to deconstruct nitrofurans and metabolites for the bathing medication and intestine is the major tissue to decompose antibiotics for the feeding medicaton. In this research, we built a completed way to determine nitrofurans and corresponding metatbolites. Comparation of fish in different conditions and different medicative ways were in this investigation. These results could be helpful for aquacultures and government institutions.

I hypothesise that cancer cells with high aldehyde dehydrogenase (ALDHhigh) activity present a new therapeutic target and will be selectively sensitive to 5-nitrofuran pro-drugs. Cancers are heterogeneous and contain subpopulations of ALDHhigh cells with tumour initiating potential. ALDH enzymes metabolize toxic aldehydes, and are highly expressed in somatic and cancer stem cells (CSCs), although their function in CSCs is not fully understood. In a small molecule screen coupled with target ID, Zhou et al. (2012) recently discovered that clinically active 5-nitrofurans (5-NFNs) are substrates of ALDH2. 5-NFNs are a class of pro-drug widely used to treat bacterial and parasitic infections, where their relative specificity is driven by nitroreductases, but little is known about the enzymes that bio-activate 5-NFNs in humans. Recent clinical cancer research has found that the 5-NFN, nifurtimox, has anti-cancer properties and it is currently in Phase 2 clinical trials for neuroblastoma and medulloblastoma (ClinicalTrials.gov Identifier: NCT00601003), however the mechanism underlying this anti-cancer activity is unknown. In melanoma and other cancers, ALDH1A1 and ALDH1A3 are highly expressed in CSCs. I demonstrate the anti-cancer activity of 5-NFNs in cancer cell lines, where they express high sensitivity to 5-NFNs in cell viability assays (A375 melanoma cells EC50 = 867nM). To test if ALDH1 enzymes are substrates of 5-NFNs, I performed in vitro activity assays by monitoring NADH production (λ = 340nm). I found that the clinically available 5-NFNs, nifuroxazide and nifurtimox, in addition to our own newly synthesised 5-NFNs, are competitive substrates for human ALDH1A3 activity in vitro (P < 0.05). Notably, nifuroxazide is not a substrate for ALDH2, suggesting that nifuroxazide may show selectivity toward ALDH1. Enzymatic assays with purified human ALDH2, demonstrate that ALDH2 requires NAD+ for bio-activation of 5-NFNs. Consistent with these assays, I found that 5-NFNs are competitive substrates for ALDH activity in melanoma cells by Aldefluor™, with 5-NFNs displaying a prolonged competitive inhibition of ALDH activity compared with the known inhibitor, DEAB. Importantly, no-nitro control compounds show no activity toward ALDH enzymes in vitro or in culture. Kinetic living-cell imaging (IncuCyte ZOOM®) reveals that a subpopulation of ALDH1A3 siRNA transfected A375 cells are protected from 5-NFN toxicity (P > 0.05) and cell death (DRAQ7™: P < 0.0001), demonstrating a functional role for ALDH1A3 in mediating 5-NFN activity in cancer cells. In contrast, A375 cells overexpressing ALDH1A3 by cDNA transient transfection were hypersensitive to 5-NFNs (P < 0.001), determined by Muse™ cell viability. Computational docking studies reveal that 5-NFNs have the potential to fit within the interior of the ALDH enzymatic cavity and interact with the catalytic cysteine, thereby offering a potential mechanism for 5-NFN bio-activation. Finally, in collaboration, we show a unique interaction between 5-NFNs and ALDH using mass spectrometry and have initiated protein crystallography trials. My work demonstrates a novel and biologically relevant 5-NFN-ALDH interaction in cancer cells. I propose 5-NFNs have the potential to target ALDHhigh CSCs within a tumour and advance the repurposing of clinical 5-NFN pro-drug antibiotics as anti-cancer therapeutics.

A surgimento de bactérias multirresistentes é uma ameaça global. Essas bactérias têm sido associadas com infecções hospitalares, no entanto, diversos casos de infecções multirresistentes adquiridas na comunidade vêm sendo relatadas, o que acendo um alerta quanto a propagação destes micro-organismos para além do ambiente hospitalar. Em todo o mundo o consumo indiscriminado de antibióticos tem aumentado significativamente, sendo este o principal fator para o surgimento e propagação de novas formas de resistência. Outro fator preocupante é a velocidade com que estas novas formas de resistência cruzam fronteiras internacionais se disseminando facilmente em todo o mundo. Este fato tem preocupado líderes mundiais, os quais consideram o aparecimento de \"superbactérias\" um pesadelo, o que pode vir a ser em um futuro próximo uma catástrofe mundial. Diante desse quadro preocupante, a necessidade de desenvolvimento de novos agentes antimicrobianos para combater essas infecções se torna iminente. Neste contexto, os compostos nitrofurânicos tem se destacado por sua atividade contra bactérias com caráter de multirresistência. Por isto, este trabalho teve como objetivo o desenvolvimento de novos compostos nitrofurânicos, tendo como composto-protótipo a nifuroxazida. O processo de desenvolvimento das estruturas análogas a nifuroxazida foi realizado a partir do planejamento da série de compostos 5-nitro-2-furfurilidênicos, com base nos parâmetros estabelecidos por Lipinski para obtenção de compostos com características farmacocinéticas e farmacodinâmicas que favorecem a biodisponibilidade, com vistas a administração por via oral. Os ensaios de avaliação da atividade antimicrobiana dos compostos foram realizados com base no método de teste de sensibilidade a agentes antimicrobianos por diluição para bactérias de crescimento aeróbio, norma M07-A9, e como base no método de padrões de crescimento para teste de suscetibilidade a antimicrobianos, vigésimo terceiro suplemento informativo, M100-S23, aprovados pelo Clinical and Laboratory Standards Institute. Os resultados de avaliação da atividade antimicrobiana indicam que o composto mais ativo c24, 6-amino-N\'-((5-nitrofurano-2-il)metileno)-2-naftohidrazida, teve atividade significativa frente a todas cepas, sendo superior a nifuroxazida, NF; nitrofurantoína, NTF; oxacilina, OXA; e vancomicina, VAN; [ (Staphylococcus aureus ATCC 29213, c24 - IC90 = 0.31 &#181;M ± 0.06; NF - IC90 = 2.39 &#181;M ± 0.08; NTF - IC90 = 5.26 &#181;M ± 0.39; OXA - IC90 = 1.14 &#181;M ± 0.18; VAN - IC90 = 0.31 &#181;M ± 0.06); (Staphylococcus aureus resistente à oxacilina, c24 - IC90 = 0.52 &#181;M ± 0.26; NF - IC90 = 5.37 &#181;M ± 0.67; NTF - IC90 = 8.20 &#181;M ± 1.66; OXA - IC90 = s. a.; VAN - IC90 = 0.50 &#181;M ± 0.20); (Staphylococcus aureus cepa heterogênea com resistência intermediária à vancomicina, hVISA-FCFHV36, c24 - IC90 = 0.82 &#181;M ± 0.07; NF - IC90 = 7.22 &#181;M ± 0.29; NTF - IC90 = 13.14 &#181;M ± 0.94; OXA - IC90 = s. a.; VAN - IC90 = 0.88 &#181;M ± 0.05); (Staphylococcus epidermidis com perfil de resistência a linezolida, c24 - IC90 = 0.74 &#181;M ± 0.02; NF - IC90 = 4.36 &#181;M ± 0.54; NTF - IC90 = 8.46 &#181;M ± 0.60; OXA - IC90 = 12.66 &#181;M ± 0.36; VAN - IC90 = 1.40 &#181;M ± 0.28); e (Enterococcus faecalis resistente à vancomicina fenótipo vanA, c24 - IC90 = 0.72 &#181;M ± 0.02; NF - IC90 = 5.09 &#181;M ± 0.08; NTF - IC90 = 9.28 &#181;M ± 0.32; OXA - IC90 = 12.26 &#181;M ± 0.72, VAN - IC90 = s. a.) ]. Entre as propriedades de maior influência na atividade do composto c24 estão ClogP e PSA. / The emergence of multidrug-resistant bacteria is a global threat. These bacteria have been associated with nosocomial infections, however, many cases of community-acquired multidrug resistant infections have been reported which light a warning about the spread of these microorganisms other than the hospital environment. Worldwide the indiscriminate use of antibiotics has increased significantly, which is the main factor for the emergence and spread of new forms of resistance. Another factor of concern is the speed with which these new forms of resistance cross international boundaries easily spreading worldwide. This has worried world leaders, who consider the emergence of \"superbugs\" a nightmare, which might be in the near future a global catastrophe. Faced with this alarming situation, the need for development of new antimicrobial agents to combat such infections becomes imminent. In this context, nitrofuran compounds has been noted for its activity against bacteria with multidrug resistance character. Therefore, this study aimed to develop new nitrofuran compounds, with the composite prototype to nifuroxazida. The process of developing similar structures nifuroxazida was performed using the designed series of substituted 5-nitro-2-furfurilidênicos, based on parameters set by Lipinski to obtain compounds with pharmacokinetic and pharmacodynamic properties that promote the bioavailability in order administration orally. The evaluation of the antimicrobial activity tests of the compounds were carried out based on the method of the antimicrobial susceptibility testing by dilution for bacterial aerobic growth, M07-A9 standard, and based on the method of growth standards for antimicrobial susceptibility testing , twenty-third informational supplement M100-S23, approved by the Clinical and Laboratory Standards Institute. The results of evaluation of the antimicrobial activity indicate that the compound more active c24, 6-amino-N\'-((5-nitrofuran-2-yl) methylene)-2-naftohidrazida had significant activity against all strains, exceeding nifuroxazida, NF; nitrofurantoin, NTF; oxacillin, OXA; and vancomycin, VAN; [(Staphylococcus aureus ATCC 29213, c24 - IC90 = 0:31 uM ± 12:06; NF - IC90 = 2.39 uM ± 12:08; NTF - IC90 = 5.26 uM ± 12:39; OXA - IC90 = 1.14 uM ± 00:18; VAN - IC90 = 0:31 uM ± 12:06); (Staphylococcus aureus resistant to methicillin, c24 - IC90 = 0:52 uM ± 00:26; NF - IC90 = 5:37 uM ± 0.67; NTF - IC90 = 8.20 uM ± 1.66; OXA - IC90 = s to .; VAN - IC90 = 0:50 uM ± 00:20.); (Staphylococcus aureus heterogeneous strain with intermediate resistance to vancomycin, hVISA-FCFHV36, c24 - IC90 = 0.82 uM ± 00:07; NF - IC90 = 7.22 uM ± 00:29; NTF - IC90 = 13:14 uM ± 0.94; OXA - IC90 = sa; VAN - IC90 = 0.88 ± 0.05 uM); (Staphylococcus epidermidis with linezolid resistance profile, c24 - IC90 = 0.74 uM ± 12:02; NF - IC90 = 4:36 uM ± 00:54; NTF - IC90 = 8:46 uM ± 0.60; OXA - IC90 = 12.66 uM ± 12:36; VAN - IC90 = uM 1:40 ± 0:28); and (Enterococcus faecalis Vancomycin-resistant vanA phenotype, c24 - IC90 = 0.72 uM ± 00:02; NF - IC90 = 9.5 uM ± 12:08; NTF - IC90 = 9.28 uM ± 00:32; OXA - IC90 = 12.26 uM ± 0.72, VAN - IC90 = sa)]. Among the most influential properties in the activity of the compound c24 are ClogP and PSA

A surgimento de bactérias multirresistentes é uma ameaça global. Essas bactérias têm sido associadas com infecções hospitalares, no entanto, diversos casos de infecções multirresistentes adquiridas na comunidade vêm sendo relatadas, o que acendo um alerta quanto a propagação destes micro-organismos para além do ambiente hospitalar. Em todo o mundo o consumo indiscriminado de antibióticos tem aumentado significativamente, sendo este o principal fator para o surgimento e propagação de novas formas de resistência. Outro fator preocupante é a velocidade com que estas novas formas de resistência cruzam fronteiras internacionais se disseminando facilmente em todo o mundo. Este fato tem preocupado líderes mundiais, os quais consideram o aparecimento de \"superbactérias\" um pesadelo, o que pode vir a ser em um futuro próximo uma catástrofe mundial. Diante desse quadro preocupante, a necessidade de desenvolvimento de novos agentes antimicrobianos para combater essas infecções se torna iminente. Neste contexto, os compostos nitrofurânicos tem se destacado por sua atividade contra bactérias com caráter de multirresistência. Por isto, este trabalho teve como objetivo o desenvolvimento de novos compostos nitrofurânicos, tendo como composto-protótipo a nifuroxazida. O processo de desenvolvimento das estruturas análogas a nifuroxazida foi realizado a partir do planejamento da série de compostos 5-nitro-2-furfurilidênicos, com base nos parâmetros estabelecidos por Lipinski para obtenção de compostos com características farmacocinéticas e farmacodinâmicas que favorecem a biodisponibilidade, com vistas a administração por via oral. Os ensaios de avaliação da atividade antimicrobiana dos compostos foram realizados com base no método de teste de sensibilidade a agentes antimicrobianos por diluição para bactérias de crescimento aeróbio, norma M07-A9, e como base no método de padrões de crescimento para teste de suscetibilidade a antimicrobianos, vigésimo terceiro suplemento informativo, M100-S23, aprovados pelo Clinical and Laboratory Standards Institute. Os resultados de avaliação da atividade antimicrobiana indicam que o composto mais ativo c24, 6-amino-N\'-((5-nitrofurano-2-il)metileno)-2-naftohidrazida, teve atividade significativa frente a todas cepas, sendo superior a nifuroxazida, NF; nitrofurantoína, NTF; oxacilina, OXA; e vancomicina, VAN; [ (Staphylococcus aureus ATCC 29213, c24 - IC90 = 0.31 &#181;M ± 0.06; NF - IC90 = 2.39 &#181;M ± 0.08; NTF - IC90 = 5.26 &#181;M ± 0.39; OXA - IC90 = 1.14 &#181;M ± 0.18; VAN - IC90 = 0.31 &#181;M ± 0.06); (Staphylococcus aureus resistente à oxacilina, c24 - IC90 = 0.52 &#181;M ± 0.26; NF - IC90 = 5.37 &#181;M ± 0.67; NTF - IC90 = 8.20 &#181;M ± 1.66; OXA - IC90 = s. a.; VAN - IC90 = 0.50 &#181;M ± 0.20); (Staphylococcus aureus cepa heterogênea com resistência intermediária à vancomicina, hVISA-FCFHV36, c24 - IC90 = 0.82 &#181;M ± 0.07; NF - IC90 = 7.22 &#181;M ± 0.29; NTF - IC90 = 13.14 &#181;M ± 0.94; OXA - IC90 = s. a.; VAN - IC90 = 0.88 &#181;M ± 0.05); (Staphylococcus epidermidis com perfil de resistência a linezolida, c24 - IC90 = 0.74 &#181;M ± 0.02; NF - IC90 = 4.36 &#181;M ± 0.54; NTF - IC90 = 8.46 &#181;M ± 0.60; OXA - IC90 = 12.66 &#181;M ± 0.36; VAN - IC90 = 1.40 &#181;M ± 0.28); e (Enterococcus faecalis resistente à vancomicina fenótipo vanA, c24 - IC90 = 0.72 &#181;M ± 0.02; NF - IC90 = 5.09 &#181;M ± 0.08; NTF - IC90 = 9.28 &#181;M ± 0.32; OXA - IC90 = 12.26 &#181;M ± 0.72, VAN - IC90 = s. a.) ]. Entre as propriedades de maior influência na atividade do composto c24 estão ClogP e PSA. / The emergence of multidrug-resistant bacteria is a global threat. These bacteria have been associated with nosocomial infections, however, many cases of community-acquired multidrug resistant infections have been reported which light a warning about the spread of these microorganisms other than the hospital environment. Worldwide the indiscriminate use of antibiotics has increased significantly, which is the main factor for the emergence and spread of new forms of resistance. Another factor of concern is the speed with which these new forms of resistance cross international boundaries easily spreading worldwide. This has worried world leaders, who consider the emergence of \"superbugs\" a nightmare, which might be in the near future a global catastrophe. Faced with this alarming situation, the need for development of new antimicrobial agents to combat such infections becomes imminent. In this context, nitrofuran compounds has been noted for its activity against bacteria with multidrug resistance character. Therefore, this study aimed to develop new nitrofuran compounds, with the composite prototype to nifuroxazida. The process of developing similar structures nifuroxazida was performed using the designed series of substituted 5-nitro-2-furfurilidênicos, based on parameters set by Lipinski to obtain compounds with pharmacokinetic and pharmacodynamic properties that promote the bioavailability in order administration orally. The evaluation of the antimicrobial activity tests of the compounds were carried out based on the method of the antimicrobial susceptibility testing by dilution for bacterial aerobic growth, M07-A9 standard, and based on the method of growth standards for antimicrobial susceptibility testing , twenty-third informational supplement M100-S23, approved by the Clinical and Laboratory Standards Institute. The results of evaluation of the antimicrobial activity indicate that the compound more active c24, 6-amino-N\'-((5-nitrofuran-2-yl) methylene)-2-naftohidrazida had significant activity against all strains, exceeding nifuroxazida, NF; nitrofurantoin, NTF; oxacillin, OXA; and vancomycin, VAN; [(Staphylococcus aureus ATCC 29213, c24 - IC90 = 0:31 uM ± 12:06; NF - IC90 = 2.39 uM ± 12:08; NTF - IC90 = 5.26 uM ± 12:39; OXA - IC90 = 1.14 uM ± 00:18; VAN - IC90 = 0:31 uM ± 12:06); (Staphylococcus aureus resistant to methicillin, c24 - IC90 = 0:52 uM ± 00:26; NF - IC90 = 5:37 uM ± 0.67; NTF - IC90 = 8.20 uM ± 1.66; OXA - IC90 = s to .; VAN - IC90 = 0:50 uM ± 00:20.); (Staphylococcus aureus heterogeneous strain with intermediate resistance to vancomycin, hVISA-FCFHV36, c24 - IC90 = 0.82 uM ± 00:07; NF - IC90 = 7.22 uM ± 00:29; NTF - IC90 = 13:14 uM ± 0.94; OXA - IC90 = sa; VAN - IC90 = 0.88 ± 0.05 uM); (Staphylococcus epidermidis with linezolid resistance profile, c24 - IC90 = 0.74 uM ± 12:02; NF - IC90 = 4:36 uM ± 00:54; NTF - IC90 = 8:46 uM ± 0.60; OXA - IC90 = 12.66 uM ± 12:36; VAN - IC90 = uM 1:40 ± 0:28); and (Enterococcus faecalis Vancomycin-resistant vanA phenotype, c24 - IC90 = 0.72 uM ± 00:02; NF - IC90 = 9.5 uM ± 12:08; NTF - IC90 = 9.28 uM ± 00:32; OXA - IC90 = 12.26 uM ± 0.72, VAN - IC90 = sa)]. Among the most influential properties in the activity of the compound c24 are ClogP and PSA