Early Stage Efficacy and Toxicology Screening for Antibiotics and Enzyme Inhibitors

Sarver, Jeffrey; Trendel, Jill A.; Bearss, Nicole R.; Wang, Lin; Luniwal, Amarjit; Erhardt, Paul; Viola, Ronald E.

doi:10.1177/1087057112438769

Cited by 11 publications

(8 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The relatively high MIC values measured for some Salmonella strains have raised concerns about the toxicity and safety of using MccJ25 in livestock production or for therapeutic purposes (Sarver et al, 2012). Using human keratinocytes, we found that neither cell viability nor LDH release was affected by MccJ25 at concentrations up to 400 μg/ml.…”

Section: Cytotoxicity and Hemolytic Assaysmentioning

confidence: 76%

Phenomic and genomic approaches to studying the inhibition of multiresistant Salmonella enterica by microcin J25

Said

Emond-Rhéault

Soltani

et al. 2020

Environmental Microbiology

View full text Add to dashboard Cite

Summary In livestock production, antibiotics are used to promote animal growth, control infections and thereby increase profitability. This practice has led to the emergence of multiresistant bacteria such as Salmonella, of which some serovars are disseminated in the environment. The objective of this study is to evaluate microcin J25 as an inhibitor of Salmonella enterica serovars of various origins including human, livestock and food. Among the 116 isolates tested, 37 (31.8%) were found resistant to at least one antibiotic, and 28 were multiresistant with 19 expressing the penta‐resistant phenotype ACSSuT. Microcin J25 inhibited all isolates, with minimal inhibitory concentration values ranging from 0.06 μg/ml (28.4 nM) to 400 μg/ml (189 μM). Interestingly, no cross‐resistance was found between microcin J25 and antibiotics. Multiple sequence alignments of genes encoding for the different proteins involved in the recognition and transport of microcin J25 showed that only ferric‐hydroxamate uptake is an essential determinant for susceptibility of S. enterica to microcin J25. Examination of Salmonella strains exposed to microcin J25 by transmission electronic microscopy showed for the first‐time involvement of a pore formation mechanism. Microcin J25 was a strong inhibitor of several multiresistant isolates of Salmonella and may have a great potential as an alternative to antibiotics.

show abstract

Section: Cytotoxicity and Hemolytic Assaysmentioning

confidence: 76%

Phenomic and genomic approaches to studying the inhibition of multiresistant Salmonella enterica by microcin J25

Said

Emond-Rhéault

Soltani

et al. 2020

Environmental Microbiology

View full text Add to dashboard Cite

show abstract

“…Here, activity against 3T3 cells stands as an initial determination of cytotoxicity. In drug discovery programs, it is common to employ a panel of mammalian cells when considering the potential toxicity of a compound ( Sarver et al., 2012 ). This is particularly important in Chagas disease, whereby the safety and tolerability of current drugs is an issue.…”

Section: Discussionmentioning

confidence: 99%

Development and application of a sensitive, phenotypic, high-throughput image-based assay to identify compound activity against Trypanosoma cruzi amastigotes

Sykes

Avery

2015

International Journal for Parasitology: Drugs and Drug Resistan

View full text Add to dashboard Cite

We have developed a high content 384-well, image-based assay to estimate the effect of compound treatment on Trypanosoma cruzi amastigotes in 3T3 fibroblasts. In the same well, the effect of compound activity on host cells can also be determined, as an initial indicator of cytotoxicity. This assay has been used to identify active compounds from an in-house library of compounds with either known biological activity or that are FDA approved, and separately, from the Medicines for Malaria Venture Malaria Box collection. Active compounds were screened against T. cruzi trypomastigotes, utilising an assay developed with the viability dye resazurin. Twelve compounds with reconfirmed solid sample activity, with IC50 values of less than 10 μM and selectivity indices to T. cruzi amastigotes over 3T3 host cells of between >22 and 319 times were identified from these libraries. As 3T3 cells are contact inhibited, with limited proliferation in the assay, selective compounds of interest were profiled in a separate assay to estimate the viability of compound treated, replicating HEK293 cells. Selective compounds that were not previously reported in the literature were further profiled by extending the incubation time against amastigote infected 3T3 cells to determine if there were residual amastigotes post-treatment, important for the consideration of the exposure time required for further biological characterisation. The assay development process and the suitability of identified compounds as hit molecules for Chagas disease research are discussed.

show abstract

“…Gao and colleagues established the first molecular assay to identify the ASADH inhibitors of Streptococcus pneumoniae (SP), Vibrio cholerae (VC) and Candida albicans (CA), by screening from a fragment library containing 378 compounds diluted as 94 cocktails [ 32 ]. They further eliminated the ineffective and toxic compounds, and finally found potent and selective ASADH inhibitors subsequently [ 33 , 34 ]. At the same time, molecular modelling and docking approach was used to identify the ASADH inhibitors of Streptococcus pneumoniae and Vibrio cholerae [ 35 ].…”

Section: Discussionmentioning

confidence: 99%

IMB-XMA0038, a new inhibitor targeting aspartate-semialdehyde dehydrogenase of Mycobacterium tuberculosis

Wang

Yang

Liu

et al. 2021

Emerging Microbes & Infections

View full text Add to dashboard Cite

The emergence of drug-resistant tuberculosis (TB) constitutes a major challenge to TB control programmes. There is an urgent need to develop effective anti-TB drugs with novel mechanisms of action. Aspartate-semialdehyde dehydrogenase (ASADH) is the second enzyme in the aspartate metabolic pathway. The absence of the pathway in humans and the absolute requirement of aspartate in bacteria make ASADH a highly attractive drug target. In this study, we used ASADH coupled with Escherichia coli type III aspartate kinase (LysC) to establish a high-throughput screening method to find new anti-TB inhibitors. IMB-XMA0038 was identified as an inhibitor of Mt ASADH with an IC 50 value of 0.59 μg/mL through screening. The interaction between IMB-XMA0038 and Mt ASADH was confirmed by surface plasmon resonance (SPR) assay and molecular docking analysis. Furthermore, IMB-XMA0038 was found to inhibit various drug-resistant MTB strains potently with minimal inhibitory concentrations (MICs) of 0.25–0.5 μg/mL. The conditional mutant strain MTB:: asadh cultured with different concentrations of inducer (10 −5 or 10 −1 μg/mL pristinamycin) resulted in a maximal 16 times difference in MICs. At the same time, IMB-XMA0038 showed low cytotoxicity in vitro and vivo . In mouse model, it encouragingly declined the MTB colony forming units (CFU) in lung by 1.67 log10 dosed at 25 mg/kg for 15 days. In conclusion, our data demonstrate that IMB-XMA0038 is a promising lead compound against drug-resistant tuberculosis.

show abstract

Early Stage Efficacy and Toxicology Screening for Antibiotics and Enzyme Inhibitors

Cited by 11 publications

References 25 publications

Phenomic and genomic approaches to studying the inhibition of multiresistant Salmonella enterica by microcin J25

Phenomic and genomic approaches to studying the inhibition of multiresistant Salmonella enterica by microcin J25

Development and application of a sensitive, phenotypic, high-throughput image-based assay to identify compound activity against Trypanosoma cruzi amastigotes

IMB-XMA0038, a new inhibitor targeting aspartate-semialdehyde dehydrogenase of Mycobacterium tuberculosis

Contact Info

Product

Resources

About