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A high throughput screen identifies potent and selective inhibitors to human epithelial 15-lipoxygenase-2.

Jameson JB, Kantz A, Schultz L, Kalyanaraman C, Jacobson MP, Maloney DJ, Jadhav A, Simeonov A, Holman TR - PLoS ONE (2014)

Bottom Line: MLS000545091 is a mixed-type inhibitor of 15-LOX-2 with a Ki of 0.9+/-0.4 µM and has a 20-fold selectivity over 5-LOX, 12-LOX, 15-LOX-1, COX-1, and COX-2.MLS000536924 is a competitive inhibitor with a Ki of 2.5+/-0.5 µM and also possesses 20-fold selectivity toward 15-LOX-2 over the other oxygenases, listed above.Finally, neither compound possesses reductive activity towards the active-site ferrous ion.

View Article: PubMed Central - PubMed

Affiliation: Chemistry and Biochemistry Department, University of California Santa Cruz, Santa Cruz, California, United States of America.

ABSTRACT
Lipoxygenase (LOX) enzymes catalyze the hydroperoxidation of arachidonic acid and other polyunsaturated fatty acids to hydroxyeicosatetraenoic acids with varying positional specificity to yield important biological signaling molecules. Human epithelial 15-lipoxygenase-2 (15-LOX-2) is a highly specific LOX isozyme that is expressed in epithelial tissue and whose activity has been correlated with suppression of tumor growth in prostate and other epithelial derived cancers. Despite the potential utility of an inhibitor to probe the specific role of 15-LOX-2 in tumor progression, no such potent/specific 15-LOX-2 inhibitors have been reported to date. This study employs high throughput screening to identify two novel, specific 15-LOX-2 inhibitors. MLS000545091 is a mixed-type inhibitor of 15-LOX-2 with a Ki of 0.9+/-0.4 µM and has a 20-fold selectivity over 5-LOX, 12-LOX, 15-LOX-1, COX-1, and COX-2. MLS000536924 is a competitive inhibitor with a Ki of 2.5+/-0.5 µM and also possesses 20-fold selectivity toward 15-LOX-2 over the other oxygenases, listed above. Finally, neither compound possesses reductive activity towards the active-site ferrous ion.

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Related in: MedlinePlus

Discovered 15-LOX-2 Selective inhibitors.
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pone-0104094-g002: Discovered 15-LOX-2 Selective inhibitors.

Mentions: To identify novel small molecule inhibitors of 15-LOX-2, we tested a diverse collection of 107,261 compounds arrayed as dilution series (57 µM to 0.73 nM). The 15-LOX-2 qHTS assay utilized the Xylenol Orange colorimetric method for detecting the hydroperoxide reaction products of lipoxygenase as described elsewhere [14], [28]. Complete screening results have been provided in the PubChem public database under Assay Identifier 882. Following the screen and data analysis, screening hits were cherry-picked and retested in the original XO assay; further re-confirmation of inhibitory activity was performed using the orthogonal cuvette-based assay as described in Methods. The compounds shown in Figure 2 were identified as the most promising hits based on their re-confirmed activity and demonstrated selectivity in the cuvette-based lipoxygenase assay.


A high throughput screen identifies potent and selective inhibitors to human epithelial 15-lipoxygenase-2.

Jameson JB, Kantz A, Schultz L, Kalyanaraman C, Jacobson MP, Maloney DJ, Jadhav A, Simeonov A, Holman TR - PLoS ONE (2014)

Discovered 15-LOX-2 Selective inhibitors.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4128814&req=5

pone-0104094-g002: Discovered 15-LOX-2 Selective inhibitors.
Mentions: To identify novel small molecule inhibitors of 15-LOX-2, we tested a diverse collection of 107,261 compounds arrayed as dilution series (57 µM to 0.73 nM). The 15-LOX-2 qHTS assay utilized the Xylenol Orange colorimetric method for detecting the hydroperoxide reaction products of lipoxygenase as described elsewhere [14], [28]. Complete screening results have been provided in the PubChem public database under Assay Identifier 882. Following the screen and data analysis, screening hits were cherry-picked and retested in the original XO assay; further re-confirmation of inhibitory activity was performed using the orthogonal cuvette-based assay as described in Methods. The compounds shown in Figure 2 were identified as the most promising hits based on their re-confirmed activity and demonstrated selectivity in the cuvette-based lipoxygenase assay.

Bottom Line: MLS000545091 is a mixed-type inhibitor of 15-LOX-2 with a Ki of 0.9+/-0.4 µM and has a 20-fold selectivity over 5-LOX, 12-LOX, 15-LOX-1, COX-1, and COX-2.MLS000536924 is a competitive inhibitor with a Ki of 2.5+/-0.5 µM and also possesses 20-fold selectivity toward 15-LOX-2 over the other oxygenases, listed above.Finally, neither compound possesses reductive activity towards the active-site ferrous ion.

View Article: PubMed Central - PubMed

Affiliation: Chemistry and Biochemistry Department, University of California Santa Cruz, Santa Cruz, California, United States of America.

ABSTRACT
Lipoxygenase (LOX) enzymes catalyze the hydroperoxidation of arachidonic acid and other polyunsaturated fatty acids to hydroxyeicosatetraenoic acids with varying positional specificity to yield important biological signaling molecules. Human epithelial 15-lipoxygenase-2 (15-LOX-2) is a highly specific LOX isozyme that is expressed in epithelial tissue and whose activity has been correlated with suppression of tumor growth in prostate and other epithelial derived cancers. Despite the potential utility of an inhibitor to probe the specific role of 15-LOX-2 in tumor progression, no such potent/specific 15-LOX-2 inhibitors have been reported to date. This study employs high throughput screening to identify two novel, specific 15-LOX-2 inhibitors. MLS000545091 is a mixed-type inhibitor of 15-LOX-2 with a Ki of 0.9+/-0.4 µM and has a 20-fold selectivity over 5-LOX, 12-LOX, 15-LOX-1, COX-1, and COX-2. MLS000536924 is a competitive inhibitor with a Ki of 2.5+/-0.5 µM and also possesses 20-fold selectivity toward 15-LOX-2 over the other oxygenases, listed above. Finally, neither compound possesses reductive activity towards the active-site ferrous ion.

Show MeSH
Related in: MedlinePlus