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      Antibacterial activity of novel dual bacterial DNA type II topoisomerase inhibitors

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          Abstract

          In this study, a drug discovery programme that sought to identify novel dual bacterial topoisomerase II inhibitors (NBTIs) led to the selection of six optimized compounds. In enzymatic assays, the molecules showed equivalent dual-targeting activity against the DNA gyrase and topoisomerase IV enzymes of Staphylococcus aureus and Escherichia coli. Consistently, the compounds demonstrated potent activity in susceptibility tests against various Gram -positive and Gram-negative reference species, including ciprofloxacin-resistant strains. The activity of the compounds against clinical multidrug-resistant isolates of S. aureus, Clostridium difficile, Acinetobacter baumannii, Neisseria gonorrhoeae, E. coli and vancomycin-resistant Enterococcus spp. was also confirmed. Two compounds ( 1 and 2) were tested in time-kill and post-antibiotic effect (PAE) assays. Compound 1 was bactericidal against all tested reference strains and showed higher activity than ciprofloxacin, and compound 2 showed a prolonged PAE, even against the ciprofloxacin-resistant S. aureus BAA-1720 strain. Spontaneous development of resistance to both compounds was selected for in S. aureus at frequencies comparable to those obtained for quinolones and other NBTIs. S. aureus BAA-1720 mutants resistant to compounds 1 and 2 had single point mutations in gyrA or gyrB outside of the quinolone resistance-determining region (QRDR), confirming the distinct site of action of these NBTIs compared to that of quinolones. Overall, the very good antibacterial activity of the compounds and their optimizable in vitro safety and physicochemical profile may have relevant implications for the development of new broad-spectrum antibiotics.

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          Challenges of antibacterial discovery.

          The discovery of novel small-molecule antibacterial drugs has been stalled for many years. The purpose of this review is to underscore and illustrate those scientific problems unique to the discovery and optimization of novel antibacterial agents that have adversely affected the output of the effort. The major challenges fall into two areas: (i) proper target selection, particularly the necessity of pursuing molecular targets that are not prone to rapid resistance development, and (ii) improvement of chemical libraries to overcome limitations of diversity, especially that which is necessary to overcome barriers to bacterial entry and proclivity to be effluxed, especially in Gram-negative organisms. Failure to address these problems has led to a great deal of misdirected effort.
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            Acinetobacter baumannii: epidemiology, antimicrobial resistance, and treatment options.

            Multidrug-resistant Acinetobacter baumannii is recognized to be among the most difficult antimicrobial-resistant gram-negative bacilli to control and treat. Increasing antimicrobial resistance among Acinetobacter isolates has been documented, although definitions of multidrug resistance vary in the literature. A. baumannii survives for prolonged periods under a wide range of environmental conditions. The organism causes outbreaks of infection and health care-associated infections, including bacteremia, pneumonia, meningitis, urinary tract infection, and wound infection. Antimicrobial resistance greatly limits the therapeutic options for patients who are infected with this organism, especially if isolates are resistant to the carbapenem class of antimicrobial agents. Because therapeutic options are limited for multidrug-resistant Acinetobacter infection, the development or discovery of new therapies, well-controlled clinical trials of existing antimicrobial regimens and combinations, and greater emphasis on the prevention of health care-associated transmission of multidrug-resistant Acinetobacter infection are essential.
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              Survey of infections due to Staphylococcus species: frequency of occurrence and antimicrobial susceptibility of isolates collected in the United States, Canada, Latin America, Europe, and the Western Pacific region for the SENTRY Antimicrobial Surveillance Program, 1997-1999.

              Between January 1997 and December 1999, bloodstream isolates from 15,439 patients infected with Staphylococcus aureus and 6350 patients infected with coagulase-negative Staphylococcus species (CoNS) were referred by SENTRY-participating hospitals in the United States, Canada, Latin America, Europe, and the Western Pacific region. S. aureus was found to be the most prevalent cause of bloodstream infection, skin and soft-tissue infection, and pneumonia in almost all geographic areas. A notable increase in methicillin (oxacillin) resistance among community-onset and hospital-acquired S. aureus strains was observed in the US centers. The prevalence of methicillin (oxacillin)-resistant S. aureus varied greatly by region, site of infection, and whether the infection was nosocomial or community onset. Rates of methicillin resistance were extremely high among S. aureus isolates from centers in Hong Kong and Japan. Uniformly high levels of methicillin resistance were observed among CoNS isolates. Given the increasing multidrug resistance among staphylococci and the possible emergence of vancomycin-resistant strains, global strategies are needed to control emergence and spread of multiply resistant staphylococci.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Project administrationRole: Writing – original draft
                Role: Investigation
                Role: Resources
                Role: Investigation
                Role: Writing – original draft
                Role: Resources
                Role: Resources
                Role: Resources
                Role: Writing – review & editing
                Role: ConceptualizationRole: SupervisionRole: Writing – original draftRole: Writing – review & editing
                Role: ConceptualizationRole: Writing – original draft
                Role: Conceptualization
                Role: Supervision
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                19 February 2020
                2020
                : 15
                : 2
                : e0228509
                Affiliations
                [1 ] Angelini RR&D (Regulatory, Research & Development)–Angelini S.p.A., S. Palomba-Pomezia (Rome), Italy
                [2 ] Department of Life and Environmental Sciences, Polytechnic University of Marche, Ancona, Italy
                University of Nebraska Medical Center, UNITED STATES
                Author notes

                Competing Interests: The authors have read the journal's policy and the authors of this manuscript have the following competing interests: NDA, ACJ, GaM, RO, MV, CB, GM, CM, FPDG and ST are employees of Angelini S.p.A. LDS reports a grant in the context of the EUREKA project co-funded by Angelini S.p.A. and by Regione Marche, Universita Politecnica delle Marche (UNIVPM - Decreti Rettorali no. 740 July 2013 and no. 812 September 2013). The authors would like to declare the following patents/patent applications associated with this research: ACJ, GaM, RO and GM are inventors in patent WO2017/211759Al pending. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

                Author information
                http://orcid.org/0000-0003-0507-5717
                Article
                PONE-D-19-21722
                10.1371/journal.pone.0228509
                7029851
                32074119
                53c9bb08-6da9-4290-9d17-f2e04d4f1825
                © 2020 D’Atanasio et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 1 August 2019
                : 16 January 2020
                Page count
                Figures: 3, Tables: 5, Pages: 21
                Funding
                Funded by: EUREKA
                Award ID: UNIVPM – Decreti Rettorali no. 740 July 2013 and no. 812 September 2013
                Award Recipient :
                This work was funded by Angelini S.p.A. Angelini S.p.A. provided support in the form of salaries for NDA, ACJ, GM, RO, MV, CB, GM, CM, FPDG and ST. LDS was supported by a grant in the context of the EUREKA project co-funded by Angelini S.p.A. and by Regione Marche, Universita Politecnica delle Marche (UNIVPM - Decreti Rettorali no. 740 July 2013 and no. 812 September 2013). The specific roles of these authors are articulated in the "author contributions" section. The funder had a role in the study design, data collection and analysis, decision to publish and preparation of the manuscript through the provision of expert decision-making input from the preclinical management teams.
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