High-Density Miniaturized Thermal Shift Assays as a General Strategy for Drug Discovery

Pantoliano, Michael W.; Petrella, Eugene C.; Kwasnoski, Joseph D.; Lobanov, Victor S.; Myslik, James C.; Graf, E. R.; Carver, Ted; Asel, Eric; Springer, Barry A.; Lane, Pamela; Salemme, F. R.

doi:10.1177/108705710100600609

Cited by 788 publications

(542 citation statements)

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“…This assay was used to screen the GSK proprietary collection of compounds and the hits identified were clustered by chemical similarity, screened to ensure no interference with the coupling reaction, and tested for stabilizing LDHA in a thermal shift stability assay [22]. From this campaign, we identified 3-((3-carbamoyl-7-(3,5-dimethylisoxazol-4-yl)-6-methoxyquinolin-4-yl)amino)benzoic acid as an NADH-competitive LDHA inhibitor.…”

Section: Resultsmentioning

confidence: 99%

Quinoline 3-sulfonamides inhibit lactate dehydrogenase A and reverse aerobic glycolysis in cancer cells

et al. 2013

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BackgroundMost normal cells in the presence of oxygen utilize glucose for mitochondrial oxidative phosphorylation. In contrast, many cancer cells rapidly convert glucose to lactate in the cytosol, a process termed aerobic glycolysis. This glycolytic phenotype is enabled by lactate dehydrogenase (LDH), which catalyzes the inter-conversion of pyruvate and lactate. The purpose of this study was to identify and characterize potent and selective inhibitors of LDHA.MethodsHigh throughput screening and lead optimization were used to generate inhibitors of LDHA enzymatic activity. Effects of these inhibitors on metabolism were evaluated using cell-based lactate production, oxygen consumption, and 13C NMR spectroscopy assays. Changes in comprehensive metabolic profile, cell proliferation, and apoptosis were assessed upon compound treatment.Results3-((3-carbamoyl-7-(3,5-dimethylisoxazol-4-yl)-6-methoxyquinolin-4-yl) amino) benzoic acid was identified as an NADH-competitive LDHA inhibitor. Lead optimization yielded molecules with LDHA inhibitory potencies as low as 2 nM and 10 to 80-fold selectivity over LDHB. Molecules in this family rapidly and profoundly inhibited lactate production rates in multiple cancer cell lines including hepatocellular and breast carcinomas. Consistent with selective inhibition of LDHA, the most sensitive breast cancer cell lines to lactate inhibition in hypoxic conditions were cells with low expression of LDHB. Our inhibitors increased rates of oxygen consumption in hepatocellular carcinoma cells at doses up to 3 microM, while higher concentrations directly inhibited mitochondrial function. Analysis of more than 500 metabolites upon LDHA inhibition in Snu398 cells revealed that intracellular concentrations of glycolysis and citric acid cycle intermediates were increased, consistent with enhanced Krebs cycle activity and blockage of cytosolic glycolysis. Treatment with these compounds also potentiated PKM2 activity and promoted apoptosis in Snu398 cells.ConclusionsRapid chemical inhibition of LDHA by these quinoline 3-sulfonamids led to profound metabolic alterations and impaired cell survival in carcinoma cells making it a compelling strategy for treating solid tumors that rely on aerobic glycolysis for survival.

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Section: Resultsmentioning

confidence: 99%

Quinoline 3-sulfonamides inhibit lactate dehydrogenase A and reverse aerobic glycolysis in cancer cells

et al. 2013

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“…Tools like "MED-SuMo" which detects surface properties of macromolecules and finds similarities among molecules [29], "CAVER" which analyzes the various tunnels and channels in the proteins [30], "FINDSITE"and "SiteMap" identifies the binding sites in the proteins (target and drug) [31,32].…”

Section: Binding Site Predictionmentioning

confidence: 99%

Bioinformatics in Drug Development

Nt¹

2017

Bioceram Dev Appl

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Bioinformatics in Drug DevelopmentKhan NT* Department of Biotechnology, Division of Life Sciences and Informatics, Balochistan University of Information Technology Engineering and Management Sciences (BUITEMS), Quetta, Pakistan AbstractIn today's world, where faster development of a new drug is crucial, not just for the patients but also for the pharmaceutical companies that are always in a competition for delivering a "new chemical entity (NCE)" to the market and the public. Bioinformatics, through various databases, web services, software and tools, has made a huge impact on the drug development process.

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“…This shift in unfolding temperature can be detected using sensitive dyes. A CCD camera was used for proper detection of the output [14].…”

Section: Detection Based On Protein Confirmationsmentioning

confidence: 99%

Molecular Nanobiotechnological Approaches for the Detection and Therapy of Prion Related Diseases

P¹,

Thampy²,

Venugopalan³

et al. 2012

Nano BioMed ENG

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Prion diseases are associated with the accumulation in the brain of an abnormal, protease resistant isoform of a host encoded glycoprotein known as prion protein (PrP). Nanotechnology in combination with biotechniques promises a broad spectrum of highly innovative approaches for overcoming the challenges posed by the prions. Recent advances in molecular nanobiotechnology have brought in the potential of molecular targeting in diagnosis and therapies of various diseases. Their high binding sensitivity and specificity added by their small size have favored the identification by in vitro protocols. Molecular targeting has initiated exciting technologies based on conjugation of biomolecules to nanoparticles. This review article is an extensive study of various research oriented nanobiotechnological protocols for rapid identification and cure for prion diseases both at in vivo and in vitro options.

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High-Density Miniaturized Thermal Shift Assays as a General Strategy for Drug Discovery

Cited by 788 publications

References 46 publications

Quinoline 3-sulfonamides inhibit lactate dehydrogenase A and reverse aerobic glycolysis in cancer cells

Quinoline 3-sulfonamides inhibit lactate dehydrogenase A and reverse aerobic glycolysis in cancer cells

Bioinformatics in Drug Development

Molecular Nanobiotechnological Approaches for the Detection and Therapy of Prion Related Diseases

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