The antifungal activity evaluation of new 2-((4-chlorophenoxy) methyl )-N- (arylcarbamothioyl) benzamides

Disciplina de Chimie farmaceutică, Facultatea de Farmacie , Universitatea de Medicină și Farmacie “Carol Davila”, București, România, Departamentul de Microbiologie, Facultatea de Biologie, Universitatea din București, București, RomâniaObiectivul studiului: Au fost proiectaţi, obţinuţi şi caracterizaţi noi derivați ai tioureei, în scopul evaluării activităţii antifungice. Materiale şi metode: Acidul 2-(4-clorofenoximetil)benzoic a fost refluxat cu clorură de tionil, obţinându-se clorura de 2-(4-clorofenoximetil)benzoil, care a reacționat cu tiocianatul de amoniu. Izotiocianatul de 2-(4-clorofenoximetil) benzoil rezultat, a fost tratat cu amine primare pentru a obţine noi derivaţi ai N-[2-(4-clorofenoximetil) benzoil]-tioureei N’-fenil substituite. Pentru evaluarea acţiunii antifungice s-au utilizat 6 tulpini de levuri izolate din surse industriale, 9 tulpini de levuri izolate din mediul clinic şi 11 tulpini de fungi filamentoşi izolaţi în urma procesului de control microbiologic al unor produse alimentare. Activitatea antifungică s-a testat calitativ, printr-o metodă difuzimetrică adaptată. Analiza cantitativă s-a realizat prin metoda microdiluţiilor seriale binare în mediu lichid. Studiul efectului sinergic al unor antifungice cu benzamidele nou sintetizate s-a realizat prin metoda E-Test. Rezultate: În urma screening-ului calitativ al activităţii antifungice a noilor bezamide testate pe levuri, s-a observat că eficienţa acestora a variat în funcţie de specia testată şi de compus. Dintre tulpinile industriale, cea mai sensibilă specie levurică s-a dovebit a fi Debariomyces hansenii. Sensibile au fost şi majoritatea tulpinilor clinice de Candida albicans. În metoda calitativă de determinare a activităţii noilor compuşi asupra tulpinilor de fungi filamentoşi, s-au observat modificări ale caracterelor de cultură, în sensul apariţiei unor colonii de dimensiuni mai reduse, ca urmare a afectării gradului de dezvoltare şi de maturare a hifelor miceliene. În determinarea cantitativă a activităţii antifungice prin stabilirea concentraţiei minime inhibitorii (CMI), în cazul levurilor, rezultate moderat vizibile s-au observat la unele specii de Candida albicans, compuşii cei mai activi fiind selectaţi pentru testarea efectului sinergic cu antifungice standard, prin metoda E-Test. În cazul testării pe tulpinile fungice de Aspergillus niger, pentru determinarea valorii CMI, s-a observat faptul că, în prezenţa unora dintre compuşi, la concentraţii mai mari, are loc persistenţa miceliului primar şi întârzierea fenomenului de maturare a miceliului secundar şi de sporulare, precum şi apariţia de corpi micelieni modificaţi. În cazul metodei E- Test, combinarea flucitozinei cu unii dintre compuși, a crescut gradul de sensibilitate al celulelor de Candida albicans la acţiunea antimicoticului. Concluzii: Au fost sintetizaţi și caracterizaţi prin spectrometrie IR, RMN și prin analiză elementală noi derivați ai benzamidei. Rezultatele testării acţiunii antifungice sugerează că acești compuși ar putea fi folosiţi în terapeutică. Studiile au fost finanţate prin contractul 13/23.12.2013 din competiţia „N. Testemiţanu”

Repurposing anti-inflammatory drugs for fighting planktonic and biofilm growth. New carbazole derivatives based on the NSAID carprofen: synthesis, in silico and in vitro bioevaluation

IntroductionOne of the promising leads for the rapid discovery of alternative antimicrobial agents is to repurpose other drugs, such as nonsteroidal anti-inflammatory agents (NSAIDs) for fighting bacterial infections and antimicrobial resistance.MethodsA series of new carbazole derivatives based on the readily available anti-inflammatory drug carprofen has been obtained by nitration, halogenation and N-alkylation of carprofen and its esters. The structures of these carbazole compounds were assigned by NMR and IR spectroscopy. Regioselective electrophilic substitution by nitration and halogenation at the carbazole ring was assigned from H NMR spectra. The single crystal X-ray structures of two representative derivatives obtained by dibromination of carprofen, were also determined. The total antioxidant capacity (TAC) was measured using the DPPH method. The antimicrobial activity assay was performed using quantitative methods, allowing establishment of the minimal inhibitory/bactericidal/biofilm eradication concentrations (MIC/MBC/MBEC) on Gram-positive (Staphylococcus aureus, Enterococcus faecalis) and Gram-negative (Escherichia coli, Pseudomonas aeruginosa) strains. Computational assays have been performed to assess the drug- and lead-likeness, pharmacokinetics (ADME-Tox) and pharmacogenomics profiles.Results and discussionThe crystal X-ray structures of 3,8-dibromocarprofen and its methyl ester have revealed significant differences in their supramolecular assemblies. The most active antioxidant compound was 1i, bearing one chlorine and two bromine atoms, as well as the CO2Me group. Among the tested derivatives, 1h bearing one chlorine and two bromine atoms has exhibited the widest antibacterial spectrum and the most intensive inhibitory activity, especially against the Gram-positive strains, in planktonic and biofilm growth state. The compounds 1a (bearing one chlorine, one NO2 and one CO2Me group) and 1i (bearing one chlorine, two bromine atoms and a CO2Me group) exhibited the best antibiofilm activity in the case of the P. aeruginosa strain. Moreover, these compounds comply with the drug-likeness rules, have good oral bioavailability and are not carcinogenic or mutagenic. The results demonstrate that these new carbazole derivatives have a molecular profile which deserves to be explored further for the development of novel antibacterial and antibiofilm agents

N-Substituted (Hexahydro)-1<i>H</i>-isoindole-1,3(2<i>H</i>)-dione Derivatives: New Insights into Synthesis and Characterization

Novel phthalimide derivatives, namely N-(1,3-dioxoisoindolin-2-yl)-2-(2-methyl-4-oxoquinazolin-3(4H)-yl)acetamide (1a) and N-(1,3-dioxoisoindolin-2-yl)thiophene-2-carboxamide (1b), and hexahydrophthalimide derivative N-(1,3-dioxohexahydro-1H-isoindol-2(3H)-yl)-2-(2-methyl-4-oxoquinazolin-3(4H)-yl)acetamide (2), have been synthesized. The phthalimide derivatives were synthesized from phthalic anhydride and 2-(2-methyl-4-oxoquinazolin-3(4H)-yl)acetohydrazide or thiophene-2-carbohydrazide, and the hexahydrophthalimide derivative has been synthesized from hexahydrophthalic anhydride and 2-(2-methyl-4-oxoquinazolin-3(4H)-yl)acetohydrazide. The chemical structures of the compounds are elucidated by Nuclear Magnetic Resonance (NMR) and Infrared (IR) spectra. The new in vitro antioxidant activities of the obtained substances were evaluated using the DPPH method. All tested compounds showed antioxidative activity, the most active compound being 1b. Bioinformatics tools were used for the prediction of pharmacokinetics and pharmacodynamics profiles. Our results showedthat all compounds have a suitable intestinal absorption rate, good BBB and CNS permeabilities and have as molecular targets MAO B, COX-2 and NF-KB, important for antioxidant activities

Synthesis and Characterization of New N-acyl Hydrazone Derivatives of Carprofen as Potential Tuberculostatic Agents

N-acyl hydrazone (NAH) is recognized as a promising framework in drug design due to its versatility, straightforward synthesis, and attractive range of biological activities, including antimicrobial, antitumoral, analgesic, and anti-inflammatory properties. In the global context of increasing resistance of pathogenic bacteria to antibiotics, NAHs represent potential solutions for developing improved treatment alternatives. Therefore, this research introduces six novel derivatives of (EZ)-N’-benzylidene-2-(6-chloro-9H-carbazol-2-yl)propanehydrazide, synthesized using a microwave-assisted method. In more detail, we joined two pharmacophore fragments in a single molecule, represented by an NSAID-type carprofen structure and a hydrazone-type structure, obtaining a new series of NSAID-N-acyl hydrazone derivatives that were further characterized spectrally using FT-IR, NMR, and HRMS investigations. Additionally, the substances were assessed for their tuberculostatic activity by examining their impact on four strains of M. tuberculosis, including two susceptible to rifampicin (RIF) and isoniazid (INH), one susceptible to RIF and resistant to INH, and one resistant to both RIF and INH. The results of our research highlight the potential of the prepared compounds in fighting against antibiotic-resistant M. tuberculosis strains

Insights into the Microbicidal, Antibiofilm, Antioxidant and Toxicity Profile of New O-Aryl-Carbamoyl-Oxymino-Fluorene Derivatives

The unprecedented increase in microbial resistance rates to all current drugs raises an acute need for the design of more effective antimicrobial strategies. Moreover, the importance of oxidative stress due to chronic inflammation in infections with resistant bacteria represents a key factor for the development of new antibacterial agents with potential antioxidant effects. Thus, the purpose of this study was to bioevaluate new O-aryl-carbamoyl-oxymino-fluorene derivatives for their potential use against infectious diseases. With this aim, their antimicrobial effect was evaluated using quantitative assays (minimum inhibitory/bactericidal/biofilms inhibitory concentrations) (MIC/MBC/MBIC), the obtained values being 0.156–10/0.312–10/0.009–1.25 mg/mL), while some of the involved mechanisms (i.e., membrane depolarization) were investigated by flow cytometry. The antioxidant activity was evaluated by studying the scavenger capacity of DPPH and ABTS•+ radicals and the toxicity was tested in vitro on three cell lines and in vivo on the crustacean Artemia franciscana Kellog. The four compounds derived from 9H-fluoren-9-one oxime proved to exhibit promising antimicrobial features and particularly, a significant antibiofilm activity. The presence of chlorine induced an electron-withdrawing effect, favoring the anti-Staphylococcus aureus and that of the methyl group exhibited a +I effect of enhancing the anti-Candida albicans activity. The IC50 values calculated in the two toxicity assays revealed similar values and the potential of these compounds to inhibit the proliferation of tumoral cells. Taken together, all these data demonstrate the potential of the tested compounds to be further used for the development of novel antimicrobial and anticancer agents

Bioevaluation of Novel Anti-Biofilm Coatings Based on PVP/Fe3O4 Nanostructures and 2-((4-Ethylphenoxy)methyl)-N- (arylcarbamothioyl)benzamides

Novel derivatives were prepared by reaction of aromatic amines with 2-(4-ethylphenoxymethyl)benzoyl isothiocyanate, affording the N-[2-(4-ethylphenoxymethyl) benzoyl]-Nꞌ-(substituted phenyl)thiourea. Structural elucidation of these compounds was performed by IR, NMR spectroscopy and elemental analysis. The new compounds were used in combination with Fe3O4 and polyvinylpyrrolidone (PVP) for the coating of medical surfaces. In our experiments, catheter pieces were coated by Matrix Assisted Pulsed Laser Evaporation (MAPLE) technique. The microbial adherence ability was investigated in 6 multi-well plates by using culture based methods. The obtained surfaces were also assessed for their cytotoxicity with respect to osteoblast cells, by using fluorescence microscopy and MTT assay. The prepared surfaces by advanced laser processing inhibited the adherence and biofilm development ability of Staphylococcus aureus and Pseudomonas aeruginosa tested strains while cytotoxic effects on the 3T3-E1 preosteoblasts embedded in layer shaped alginate hydrogels were not observed. These results suggest that the obtained medical surfaces, based on the novel thiourea derivatives and magnetic nanoparticles with a polymeric shell could represent a promising alternative for the development of new and effective anti-infective strategies

Bioevaluation of Novel Anti-Biofilm Coatings Based on PVP/Fe3O4 Nanostructures and 2-((4-Ethylphenoxy)methyl)-N- (arylcarbamothioyl)benzamides

Novel derivatives were prepared by reaction of aromatic amines with 2-(4-ethylphenoxymethyl)benzoyl isothiocyanate, affording the N-[2-(4-ethylphenoxymethyl) benzoyl]-Nꞌ-(substituted phenyl)thiourea. Structural elucidation of these compounds was performed by IR, NMR spectroscopy and elemental analysis. The new compounds were used in combination with Fe3O4 and polyvinylpyrrolidone (PVP) for the coating of medical surfaces. In our experiments, catheter pieces were coated by Matrix Assisted Pulsed Laser Evaporation (MAPLE) technique. The microbial adherence ability was investigated in 6 multi-well plates by using culture based methods. The obtained surfaces were also assessed for their cytotoxicity with respect to osteoblast cells, by using fluorescence microscopy and MTT assay. The prepared surfaces by advanced laser processing inhibited the adherence and biofilm development ability of Staphylococcus aureus and Pseudomonas aeruginosa tested strains while cytotoxic effects on the 3T3-E1 preosteoblasts embedded in layer shaped alginate hydrogels were not observed. These results suggest that the obtained medical surfaces, based on the novel thiourea derivatives and magnetic nanoparticles with a polymeric shell could represent a promising alternative for the development of new and effective anti-infective strategies