AG205, a Novel Agent Directed against FabK of Streptococcus pneumoniae

ABSTRACT AG205 was identified from high-throughput screening as a potent inhibitor of FabK, the enoyl-ACP reductase in Streptococcus pneumoniae. Specific inhibition of lipid biosynthesis in a macromolecular biosynthesis assay and identification of an Ala141Ser substitution in FabK from spontaneous AG205-resistant mutants indicated that AG205 exerts antibacterial activity against S. pneumoniae through the specific inhibition of FabK.

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Discovery of a Novel and Potent Class of FabI-Directed Antibacterial Agents

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.46.10.3118-3124.2002 ◽

2002 ◽

Vol 46 (10) ◽

pp. 3118-3124 ◽

Cited By ~ 113

Author(s):

David J. Payne ◽

William H. Miller ◽

Valerie Berry ◽

John Brosky ◽

Walter J. Burgess ◽

...

Keyword(s):

Antibacterial Activity ◽

High Throughput ◽

High Throughput Screening ◽

Antibacterial Agents ◽

Fatty Acid Biosynthesis ◽

Acyl Carrier Protein ◽

Multidrug Resistant ◽

Infection Model ◽

Enoyl Acp Reductase

ABSTRACT Bacterial enoyl-acyl carrier protein (ACP) reductase (FabI) catalyzes the final step in each elongation cycle of bacterial fatty acid biosynthesis and is an attractive target for the development of new antibacterial agents. High-throughput screening of the Staphylococcus aureus FabI enzyme identified a novel, weak inhibitor with no detectable antibacterial activity against S. aureus. Iterative medicinal chemistry and X-ray crystal structure-based design led to the identification of compound 4 [(E)-N-methyl-N-(2-methyl-1H-indol-3-ylmethyl)-3-(7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3-yl)acrylamide], which is 350-fold more potent than the original lead compound obtained by high-throughput screening in the FabI inhibition assay. Compound 4 has exquisite antistaphylococci activity, achieving MICs at which 90% of isolates are inhibited more than 500 times lower than those of nine currently available antibiotics against a panel of multidrug-resistant strains of S. aureus and Staphylococcus epidermidis. Furthermore, compound 4 exhibits excellent in vivo efficacy in an S. aureus infection model in rats. Biochemical and genetic approaches have confirmed that the mode of antibacterial action of compound 4 and related compounds is via inhibition of FabI. Compound 4 also exhibits weak FabK inhibitory activity, which may explain its antibacterial activity against Streptococcus pneumoniae and Enterococcus faecalis, which depend on FabK and both FabK and FabI, respectively, for their enoyl-ACP reductase function. These results show that compound 4 is representative of a new, totally synthetic series of antibacterial agents that has the potential to provide novel alternatives for the treatment of S. aureus infections that are resistant to our present armory of antibiotics.

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Large-scale High-throughput Screening Revealed 5'-(carbonylamino)-2,3'- bithiophene-4'-carboxylate as Novel Template for Antibacterial Agents

Current Drug Discovery Technologies ◽

10.2174/1570163816666190603095521 ◽

2020 ◽

Vol 17 (5) ◽

pp. 716-724

Author(s):

Yan A. Ivanenkov ◽

Renat S. Yamidanov ◽

Ilya A. Osterman ◽

Petr V. Sergiev ◽

Vladimir A. Aladinskiy ◽

...

Keyword(s):

Antibacterial Activity ◽

High Throughput ◽

High Throughput Screening ◽

Large Scale ◽

Antibacterial Agents ◽

Sos Response ◽

Luciferase Assay ◽

Translational Machinery ◽

E Coli ◽

New Class

Background: The key issue in the development of novel antimicrobials is a rapid expansion of new bacterial strains resistant to current antibiotics. Indeed, World Health Organization has reported that bacteria commonly causing infections in hospitals and in the community, e.g. E. Coli, K. pneumoniae and S. aureus, have high resistance vs the last generations of cephalosporins, carbapenems and fluoroquinolones. During the past decades, only few successful efforts to develop and launch new antibacterial medications have been performed. This study aims to identify new class of antibacterial agents using novel high-throughput screening technique. Methods: We have designed library containing 125K compounds not similar in structure (Tanimoto coeff.< 0.7) to that published previously as antibiotics. The HTS platform based on double reporter system pDualrep2 was used to distinguish between molecules able to block translational machinery or induce SOS-response in a model E. coli system. MICs for most active chemicals in LB and M9 medium were determined using broth microdilution assay. Results: In an attempt to discover novel classes of antibacterials, we performed HTS of a large-scale small molecule library using our unique screening platform. This approach permitted us to quickly and robustly evaluate a lot of compounds as well as to determine the mechanism of action in the case of compounds being either translational machinery inhibitors or DNA-damaging agents/replication blockers. HTS has resulted in several new structural classes of molecules exhibiting an attractive antibacterial activity. Herein, we report as promising antibacterials. Two most active compounds from this series showed MIC value of 1.2 (5) and 1.8 μg/mL (6) and good selectivity index. Compound 6 caused RFP induction and low SOS response. In vitro luciferase assay has revealed that it is able to slightly inhibit protein biosynthesis. Compound 5 was tested on several archival strains and exhibited slight activity against gram-negative bacteria and outstanding activity against S. aureus. The key structural requirements for antibacterial potency were also explored. We found, that the unsubstituted carboxylic group is crucial for antibacterial activity as well as the presence of bulky hydrophobic substituents at phenyl fragment. Conclusion: The obtained results provide a solid background for further characterization of the 5'- (carbonylamino)-2,3'-bithiophene-4'-carboxylate derivatives discussed herein as new class of antibacterials and their optimization campaign.

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Abstract 3414: High-throughput screening identifies mithramycin as a potent inhibitor of the EWS-FLI1 transcription factor and a strong cytotoxic agent in Ewing's sarcoma cells

10.1158/1538-7445.am10-3414 ◽

2010 ◽

Author(s):

Patrick J. Grohar ◽

Girma M. Woldemichael ◽

Laurie B. Griffin ◽

Choh Yeung ◽

Qing-Rong Chen ◽

...

Keyword(s):

Transcription Factor ◽

High Throughput ◽

High Throughput Screening ◽

Ewing’S Sarcoma ◽

Ewing's Sarcoma ◽

Potent Inhibitor ◽

Cytotoxic Agent ◽

Ewing's Sarcoma Cells ◽

Sarcoma Cells

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Common features of antibacterial compounds: an analysis of 10000 compounds library

Biomeditsinskaya Khimiya ◽

10.18097/pbmc20156106785 ◽

2015 ◽

Vol 61 (6) ◽

pp. 785-790 ◽

Cited By ~ 1

Author(s):

M.S. Veselov ◽

P.V. Sergiev ◽

I.A. Osterman ◽

D.A. Skvortsov ◽

A.Ya. Golovina ◽

...

Keyword(s):

Escherichia Coli ◽

Antibacterial Activity ◽

Organic Compounds ◽

High Throughput ◽

High Throughput Screening ◽

Chemical Space ◽

Molecular Target ◽

Antibacterial Compounds ◽

Common Features ◽

Automated Screening

Antibacterial compounds are one of the essential classes of clinically important drugs. High throughput screening allowed revealing potential antibiotics active towards any molecular target in bacterial cell. We used a library of 9820 organic compounds with highly diversified structures to screen for antibacterial activity. As the result of automated screening, 103 compounds were found to possess antibacterial activity against Escherichia coli. The properties of these compounds were compared with those of initial library. Non-linear Kohonen mapping was used to analyze the differences between non-active molecules from initial library, identified antibacterial hits and compounds with reported antibacterial activity. It was found that identified antibacterial compounds are located in the separated area of chemical space. It can be therefore suggested that these molecules belong to novel classes of antibacterial compounds and could be studied further.

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Repurposing cytarabine for treating primary effusion lymphoma by targeting KSHV latent and lytic replications

10.1101/299339 ◽

2018 ◽

Author(s):

Marion Gruffaz ◽

Shenghua Zhou ◽

Karthik Vasan ◽

Teresa Rushing ◽

Qing Liu Michael ◽

...

Keyword(s):

Mouse Model ◽

High Throughput ◽

High Throughput Screening ◽

Therapeutic Agent ◽

Potent Inhibitor ◽

Primary Effusion Lymphoma ◽

Viral Dna ◽

Dna And Rna ◽

Virion Production ◽

Novel Therapeutic

AbstractOncogenic Kaposi’s sarcoma-associated herpesvirus (KSHV) is etiologically linked to primary effusion lymphoma (PEL), an aggressive and non-treatable malignancy commonly found in AIDS patients. In this study, we performed a high throughput screening of 3,731 characterized compounds, and identified cytarabine approved by FDA for treating numerous types of cancer as a potent inhibitor of KSHV-induced PEL. We showed the high efficacy of cytarabine in the growth inhibition of various PEL cells by inducing cell cycle arrest and apoptosis. Cytarabine inhibited host DNA and RNA syntheses and therefore induced cellular cytotoxicity. Furthermore, cytarabine inhibited viral DNA and RNA syntheses and induced the the rapid degradation of KSHV major latent protein LANA, leading to the suppression of KSHV latent replication. Importantly, cytarabine effectively inhibited active KSHV replication and virion production in PEL cells. Finally, cytarabine treatments not only effectively inhibited the initiation and progression of PEL tumors, but also induced regression of grown PEL tumors in a xenograft mouse model. Together, our study has identified cytarabine as novel therapeutic agent for treating PEL as well as eliminating KSHV persistent infection.ImportancePrimary effusion lymphoma is an aggressive malignancy caused by Kaposi’s sarcoma-associated herpesvirus. The outcome of primary effusion lymphoma is dismal without specific treatment. Through a high throughput screening of characterized compounds, we identified a FDA-approved compound cytarabine as a potent inhibitor of primary effusion lymphoma. We showed that cytarabine induced regression of PEL tumors in a xenograft mouse model. Cytarabine inhibited host and viral DNA and RNA syntheses, resulting in the induction of cytotoxicity. Of interest, cytarabine induced the degradation of KSHV major latent protein LANA, hence suppressing KSHV latent replication, which is required for PEL survival. Furthermore, cytarabine inhibited KSHV lytic replication program, preventing virion production. Our findings identified cytarabine as novel therapeutic agent for treating PEL as well as for eliminating KSHV persistent infection. Since cytarabine is already approved by the FDA, it might be an ideal candidate for repurposing for PEL therapy and for further evaluation in advanced clinical trials.

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High throughput screening to determine the antibacterial activity of Terminalia ferdinandiana (Kakadu plum): A proof of concept

Journal of Microbiological Methods ◽

10.1016/j.mimet.2021.106169 ◽

2021 ◽

Vol 182 ◽

pp. 106169

Author(s):

Saleha Akter ◽

Yasmina Sultanbawa ◽

Daniel Cozzolino

Keyword(s):

Antibacterial Activity ◽

High Throughput ◽

High Throughput Screening ◽

Proof Of Concept ◽

Kakadu Plum ◽

Terminalia Ferdinandiana

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Inhibitors of Streptococcus pneumoniae Surface Endonuclease EndA Discovered by High-Throughput Screening Using a PicoGreen Fluorescence Assay

CrossRef Listing of Deleted DOIs ◽

10.1177/1087057112461153 ◽

2012 ◽

Vol 18 (3) ◽

pp. 247-257 ◽

Cited By ~ 10

Author(s):

Eliza J. R. Peterson ◽

Dmitri Kireev ◽

Andrea F. Moon ◽

Marika Midon ◽

William P. Janzen ◽

...

Keyword(s):

Streptococcus Pneumoniae ◽

Small Molecules ◽

Virulence Factors ◽

High Throughput ◽

High Throughput Screening ◽

Antimicrobial Agents ◽

Pneumococcal Infection ◽

Dna Uptake ◽

Activity Data ◽

High Throughput Screening Assay

The human commensal pathogen Streptococcus pneumoniae expresses a number of virulence factors that promote serious pneumococcal diseases, resulting in significant morbidity and mortality worldwide. These virulence factors may give S. pneumoniae the capacity to escape immune defenses, resist antimicrobial agents, or a combination of both. Virulence factors also present possible points of therapeutic intervention. The activities of the surface endonuclease, EndA, allow S. pneumoniae to establish invasive pneumococcal infection. EndA’s role in DNA uptake during transformation contributes to gene transfer and genetic diversification. Moreover, EndA’s nuclease activity degrades the DNA backbone of neutrophil extracellular traps (NETs), allowing pneumococcus to escape host immune responses. Given its potential impact on pneumococcal pathogenicity, EndA is an attractive target for novel antimicrobial therapy. Herein, we describe the development of a high-throughput screening assay for the discovery of nuclease inhibitors. Nuclease-mediated digestion of double-stranded DNA was assessed using fluorescence changes of the DNA dye ligand, PicoGreen. Under optimized conditions, the assay provided robust and reproducible activity data ( Z′= 0.87) and was used to screen 4727 small molecules against an imidazole-rescued variant of EndA. In total, six small molecules were confirmed as novel EndA inhibitors, some of which may have utility as research tools for understanding pneumococcal pathogenesis and for drug discovery.

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