High throughput screening of a library based on kinase inhibitor scaffolds against Mycobacterium tuberculosis H37Rv

Reynolds, Robert C.; Ananthan, Subramaniam; Faaleolea, Ellen; Hobrath, Judith V.; Kwong, Cecil D.; Maddox, Clinton; Rasmussen, Lynn; Sosa, Melinda; Thammasuvimol, Elizabeth; White, E. Lucile; Zhang, Wei; Secrist, John A.

doi:10.1016/j.tube.2011.05.005

Cited by 87 publications

(109 citation statements)

References 57 publications

Supporting

Mentioning

107

Contrasting

Order By: Relevance

“…Intriguingly, KKL-35 was also identified in a HTS for molecules that prevent growth of Mycobacterium tuberculosis (26). Based on the data presented here, we suggest that KKL-35 kills M. tuberculosis by inhibiting trans-translation.…”

Section: Discussionsupporting

confidence: 52%

Small molecule inhibitors of trans -translation have broad-spectrum antibiotic activity

Ramadoss

Alumasa

Cheng

et al. 2013

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

View full text Add to dashboard Cite

The trans-translation pathway for protein tagging and ribosome release plays a critical role for viability and virulence in a wide range of pathogens but is not found in animals. To explore the use of trans-translation as a target for antibiotic development, a highthroughput screen and secondary screening assays were used to identify small molecule inhibitors of the pathway. Compounds that inhibited protein tagging and proteolysis of tagged proteins were recovered from the screen. One of the most active compounds, KKL-35, inhibited the trans-translation tagging reaction with an IC 50 = 0.9 μM. KKL-35 and other compounds identified in the screen exhibited broad-spectrum antibiotic activity, validating trans-translation as a target for drug development. This unique target could play a key role in combating strains of pathogenic bacteria that are resistant to existing antibiotics.antibiotic target | tmRNA | non-stop translation T he increasing prevalence of antibiotic-resistant bacterial pathogens has spurred a search for new pathways that can be targeted for antibiotic development (1, 2). One pathway that has not been exploited is the trans-translation pathway, which resolves nonstop translation complexes. The components of trans-translation have been identified in every sequenced bacterial genome, and mutations in these components affect viability or virulence in a wide range of bacteria (3, 4), suggesting that inhibitors of trans-translation might be effective broad-spectrum antibiotics. In addition, the trans-translation pathway is not found in animals, so specific inhibitors are expected to have few side effects on the host.The purpose of trans-translation is to remove nonstop translation complexes, i.e., translation reactions in which the ribosome has reached the 3′ end of the mRNA without terminating at a stop codon (4-6). These complexes are prevalent in bacteria because bacterial ribosomes do not require any information from the 3′ end of the mRNA to initiate translation, and bacteria lack most of the mechanisms for mRNA proofreading found in eukaryotes (7). Because hydrolysis of peptidyl-tRNA by release factors requires a stop codon in the A site, normal translation termination cannot occur when the ribosome reads to the 3′ end of the mRNA and there is no in-frame stop codon. Transtranslation resolves nonstop translation complexes using a ribonucleoprotein complex containing transfer-messenger RNA (tmRNA) and the small protein SmpB (4-6). tmRNA-SmpB recognizes nonstop translation complexes and enters the ribosomal A site mimicking a tRNA (8). The nascent polypeptide is transferred to tmRNA, and a specialized reading frame within tmRNA is inserted into the mRNA channel (4-6). Translation resumes using this sequence as a message and terminates at a stop codon at the end of the reading frame, releasing the ribosome and a tagged protein (4-6). The tag sequence is recognized by proteases, and the tagged protein is rapidly degraded (9-12). The net reaction of trans-translation is the removal of all components o...

show abstract

Section: Discussionsupporting

confidence: 52%

Small molecule inhibitors of trans -translation have broad-spectrum antibiotic activity

Ramadoss

Alumasa

Cheng

et al. 2013

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

View full text Add to dashboard Cite

show abstract

“…In that time, more than 9 million compounds have been screened against 36 infectious agents, at both the BSL-2 and the BSL-3 level. [9][10][11][12][13][14][15][16][17] Many assays were adapted to the automation platform and screened successfully. But there were assays that could only achieve semi-high throughput due to the technical limitations of conventional liquid handling.…”

Section: Resultsmentioning

confidence: 99%

Acoustic Droplet Ejection Applications for High-Throughput Screening of Infectious Agents

2016

Self Cite

View full text Add to dashboard Cite

When acoustic droplet ejection technology was first introduced for high-throughput applications, it was used primarily for dispensing compounds dissolved in DMSO. The high precision and accuracy achieved for low-volume transfers in this application were noted by those working outside of the compound management area, and interest was generated in expanding the scope of the technology to include other liquid types. Later-generation instruments included calibrations for several aqueous buffers that were applicable to the life sciences. The High Throughput Screening Center at Southern Research has made use of this range of liquid calibrations for the Infectious Disease Program. The original calibration for DMSO has allowed the preparation of assay-ready plates that can be sent to remote locations. This process was used as part of the collaboration between Southern Research and Galveston National Laboratory, University of Texas Medical Branch, to develop high-throughput screening for biological safety level 4 containment and to provide compounds for two pilot screens that were run there with BSL-4-level pathogens. The aqueous calibrations have been instrumental in miniaturizing assays used for infectious disease, such as qPCR, tissue culture infectious dose 50, and bacterial motility, to make them compatible with HTS operations.

show abstract

“…38 Four of the five recently identified inhibitors of trans-translation contain the core structure, 39 but these four compounds are not the same as the ones identified in this study. As noted previously, 34 'there are numerous literature examples of the 2,5-disubstituted-1,3,4-oxadiazole system that have shown biological activities including anticancer, anti-inflammatory, antifungal, antiviral, and antibacterial or antimycobacterial activity'. Amongst the compounds with this core structure that have appeared in literature, only one compound described here (LC5) has appeared in literature.…”

Section: Tablementioning

confidence: 91%

“…A component of the core structure is the 1,3,4-oxadiazole moiety. Compounds containing the 1,3,4-oxadiazole moiety have been identified in several studies (e.g., [33][34][35][36][37], and reviewed recently. 38 Four of the five recently identified inhibitors of trans-translation contain the core structure, 39 but these four compounds are not the same as the ones identified in this study.…”

Section: Tablementioning

confidence: 99%

Identification of Bacillus anthracis PurE inhibitors with antimicrobial activity

Kim

Wolf

Zhu

et al. 2015

Bioorganic & Medicinal Chemistry

View full text Add to dashboard Cite

High throughput screening of a library based on kinase inhibitor scaffolds against Mycobacterium tuberculosis H37Rv

Cited by 87 publications

References 57 publications

Small molecule inhibitors of trans -translation have broad-spectrum antibiotic activity

Small molecule inhibitors of trans -translation have broad-spectrum antibiotic activity

Acoustic Droplet Ejection Applications for High-Throughput Screening of Infectious Agents

Identification of Bacillus anthracis PurE inhibitors with antimicrobial activity

Contact Info

Product

Resources

About