Rising standards for tuberculosis drug development

Balganesh, Tanjore S.; Alzari, Pedro M.; Cole, Stewart T.

doi:10.1016/j.tips.2008.08.001

Cited by 79 publications

(52 citation statements)

References 47 publications

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“…Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB), infects approximately one third of the world's population, and 1.7-1.8 million people die from this disease annually (1). Agents that are active against Mtb are urgently needed to combat this global epidemic that is heavily influenced by resistance to the available regimen of drugs, lengthy treatment, and co-infection with HIV.…”

Section: Introductionmentioning

confidence: 99%

Validating New Tuberculosis Computational Models with Public Whole Cell Screening Aerobic Activity Datasets

Ekins

Freundlich

2011

Pharm Res

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Section: Introductionmentioning

confidence: 99%

Validating New Tuberculosis Computational Models with Public Whole Cell Screening Aerobic Activity Datasets

Ekins

Freundlich

2011

Pharm Res

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“…Although a positive signal does not provide definitive MOA assignment, the selective induction of the reporter gene nevertheless affords useful insight into the broad cellular process or metabolic pathway disrupted, irrespective of whether the specific molecular target is known (19,20). Critically, this approach provides a means of biologically profiling compounds with whole-cell activity before extensive medicinal chemistry efforts are undertaken (21,22).…”

mentioning

confidence: 99%

Bioluminescent Reporters for Rapid Mechanism of Action Assessment in Tuberculosis Drug Discovery

Naran

Moosa

Barry

et al. 2016

Antimicrob Agents Chemother

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cThe tuberculosis (TB) drug discovery pipeline is fueled by compounds identified in whole-cell screens against the causative agent, Mycobacterium tuberculosis. Phenotypic screening enables the selection of molecules that inhibit essential cellular functions in live, intact bacilli grown under a chosen in vitro condition. However, deducing the mechanism of action (MOA), which is important to avoid promiscuous targets, often requires significant biological resources in a lengthy process that risks decoupling medicinal chemistry and biology efforts. Therefore, there is a need to develop methods enabling rapid MOA assessment of putative "actives" for triage decisions. Here, we describe a modified version of a bioluminescence reporter assay that allows nondestructive detection of compounds targeting either of two macromolecular processes in M. tuberculosis: cell wall biosynthesis or maintenance of DNA integrity. Coupling the luxCDABE operon from Photorhabdus luminescens to mycobacterial promoters driving expression of the iniBAC operon (PiniB-LUX) or the DNA damage-inducible genes, recA (PrecA-LUX) or radA (PradA-LUX), provided quantitative detection in real time of compounds triggering expression of any of these promoters over an extended 10-to 12-day incubation. Testing against known anti-TB agents confirmed the specificity of each reporter in registering the MOA of the applied antibiotic in M. tuberculosis, independent of bactericidal or bacteriostatic activity. Moreover, profiles obtained for experimental compounds indicated the potential to infer complex MOAs in which multiple cellular processes are disrupted. These results demonstrate the utility of the reporters for early triage of compounds based on the provisional MOA and suggest their application to investigate polypharmacology in known and experimental anti-TB agents.

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“…Tuberculosis remains a major public health problem, and antibiotic development is being actively pursued (4,5). Using transposon mutagenesis and DNA microarray analysis, we have identified genes that are essential for the growth of Mycobacterium tuberculosis, the causative organism of tuberculosis (6).…”

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confidence: 99%

Depletion of antibiotic targets has widely varying effects on growth

Wei

Krishnamoorthy

Murphy

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

145

201

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It is often assumed that antibiotics act on the most vulnerable cellular targets, particularly those that require limited inhibition to block growth. To evaluate this assumption, we developed a genetic method that can inducibly deplete targeted proteins and that mimics their chemical inactivation. We applied this system to current antibiotic targets in mycobacteria. Although depleting some antibiotic targets significantly perturbs bacterial growth, surprisingly, we found that reducing the levels of other targets by more than 97% had little or no effect on growth. For one of these targets, dihydrofolate reductase, metabolic analysis suggested that depletion mimics the use of subinhibitory concentrations of the antibiotic trimethroprim. These observations indicate that some drug targets can exist at levels much higher than are needed to support growth. However, protein depletion can be used to identify promising drug targets that are particularly vulnerable to inhibition.inducible proteolysis | HIV protease | trimethoprim

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Rising standards for tuberculosis drug development

Cited by 79 publications

References 47 publications

Validating New Tuberculosis Computational Models with Public Whole Cell Screening Aerobic Activity Datasets

Validating New Tuberculosis Computational Models with Public Whole Cell Screening Aerobic Activity Datasets

Bioluminescent Reporters for Rapid Mechanism of Action Assessment in Tuberculosis Drug Discovery

Depletion of antibiotic targets has widely varying effects on growth

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