RESUMO
Antimicrobial resistance is a major global threat to human health due to the rise, spread and persistence of multi-drug-resistant bacteria or 'superbugs'. There is an urgent need to develop novel chemotherapeutics to overcome this overarching challenge. The authors derivatized a clinically used fluoroquinolone antibiotic ciprofloxacin (Cip), and complexed it to a copper phenanthrene framework. This resulted in the development of two novel metallo-antibiotics of general formula [Cu(N,N)(CipHA)]NO3 where N,N represents a phenanthrene ligand and CipHA represents a hydroxamic acid of Cip derivative. Comprehensive studies, including a detailed proteomic study in which Staphylococcus aureus cells were exposed to the complexes, were undertaken to gain an insight into their mode of action. These new complexes possess potent antibacterial activity against S. aureus and methicillin-resistant S. aureus. In addition, they were found to be well tolerated in vivo in Galleria mellonella larvae, which has both functional and structural similarities to the innate immune system of mammals. These findings suggest that proteins involved in virulence, pathogenesis, and the synthesis of nucleotides and DNA repair mechanisms are most affected. In addition, both complexes affected similar cell pathways when compared with clinically used Cip, including cationic antimicrobial peptide resistance. The Cu-DPPZ-CipHA (DPPZ = dipyrido[3,2-a:2',3'-c]phenazine) analogue also induces cell leakage, which leads to an altered proteome indicative of reduced virulence and increased stress.
Assuntos
Antibacterianos/farmacologia , Ciprofloxacina/análogos & derivados , Ciprofloxacina/farmacologia , Cobre/farmacologia , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Animais , Cobre/química , Desenho de Fármacos , Avaliação Pré-Clínica de Medicamentos , Farmacorresistência Bacteriana Múltipla/genética , Humanos , Ácidos Hidroxâmicos/química , Staphylococcus aureus Resistente à Meticilina/genética , Mariposas/efeitos dos fármacos , Fenantrenos/química , Fenantrenos/farmacologia , Infecções Estafilocócicas/tratamento farmacológicoRESUMO
A fast and automated separation and quantification method for bromide and the artificial nucleoside 5-bromo-2'-deoxyuridine (5-BrdU) via hyphenation of ion exchange chromatography (IC) and inductively coupled plasma-mass spectrometry (ICP-MS) is presented. The analysis of these two species is relevant to monitor the transfer of electrons along metal-mediated DNA base pairs. Charge transfer in DNA is of high interest for the implementation in nanotechnological applications like molecular wires. 5-BrdU as part of the DNA sequence releases bromide upon one electron reduction after efficient electron transfer along the DNA. The concentrations of 5-BrdU and bromide in enzymatically digested DNA samples can therefore be used as a marker for the efficiency of electron transfer along the DNA helix. A large number of samples was analyzed using an automated IC system. This platform enables time-efficient external calibration by inline dilution of a stock solution. Due to the fast separation of the two bromine species in less than 90 s, the developed method is suitable for screening applications with a multitude of samples. Despite the isobaric interferences and a low degree of ionization for bromine detection via ICP-MS the method has a limit of detection (LOD) of 30 ng/L which is approximately an order of magnitude lower than a comparable method using reversed phase high performance liquid chromatography (RP-HPLC) and ICP-MS.