A potent and selective inhibitor targeting human and murine 12/15-LOX. Issue 6 (15th March 2016)

• Record Type: Journal Article
• Authors: Armstrong, Michelle M.; Freedman, Cody J.; Jung, Joo Eun; Zheng, Yi; Kalyanaraman, Chakrapani; Jacobson, Matthew P.; Simeonov, Anton; Maloney, David J.; van Leyen, Klaus; Jadhav, Ajit; Holman, Theodore R.
• Is Part Of: Bioorganic & medicinal chemistry. Volume 24:Issue 6(2016)
• Page Start: 1183
• Page End: 1190
• Publication Date: 2016-03-15
• Subjects: LOX lipoxygenase -- hLOX human lipoxygenase -- h15-LOX-2 human epithelial 15-lipoxygenase-2 -- h12/15-LOX, h15-LOX-1 human reticulocyte 15-lipoxygenase-1 -- h12-LOX human platelet 12-lipoxygenase -- s15-LOX-1 soybean 15-lipoxygenase-1 -- h5-LOX human leukocyte 5-lipoxygenase -- AA arachidonic acid -- LA linoleic acid -- 12-HETE 12-hydroxy-5, 8, 10, 14-eicosatetraenoic acid -- 12-HpETE 12-hydroperoxyeicosatetraenoic acid -- 15-HpETE 15-hydroperoxyeicosatetraenoic acid -- 13-HpODE 13-(S)-hydroperoxyoctadecadienoic acid -- tPA tissue plasminogen activator -- AIF apoptosis-inducing factor -- HTS High Throughput Screen -- CHO Chinese hamster ovary -- MLSMR Molecular Libraries Small Molecule Repository -- 99089 MLS000099089 -- NIH National Institutes of Health -- ICP-MS inductively coupled plasma mass spectrometer -- BSA bovine serum albumin -- EDTA Ethylenediaminetetraacetic acid -- DMSO dimethyl sulfoxide -- SAR structure–activity relationship -- MCAO middle cerebral artery occlusion -- SDS–PAGE sodium dodecyl sulfate–polyacrylamide gel electrophoresis; Inhibitor -- Selective -- High-throughput -- Lipoxygenase -- Human -- Murine; Bioorganic chemistry -- Periodicals; Pharmaceutical chemistry -- Periodicals; Biochemistry -- Periodicals; Chemistry, Clinical -- Periodicals; Chemistry, Organic -- Periodicals; Chimie bio-organique -- Périodiques; Chimie pharmaceutique -- Périodiques; 615.19
• DOI: 10.1016/j.bmc.2016.01.042
• Languages: English
• ISSNs: 0968-0896

Abstract

Graphical abstract: Abstract: Human reticulocyte 12/15-lipoxygenase (h12/15-LOX) is a lipid-oxidizing enzyme that can directly oxidize lipid membranes in the absence of a phospholipase, leading to a direct attack on organelles, such as the mitochondria. This cytotoxic activity of h12/15-LOX is up-regulated in neurons and endothelial cells after a stroke and thought to contribute to both neuronal cell death and blood–brain barrier leakage. The discovery of inhibitors that selectively target recombinant h12/15-LOX in vitro, as well as possessing activity against the murine ortholog ex vivo, could potentially support a novel therapeutic strategy for the treatment of stroke. Herein, we report a new family of inhibitors discovered in a High Throughput Screen (HTS) that are selective and potent against recombinant h12/15-LOX and cellular mouse 12/15-LOX (m12/15-LOX). MLS000099089 (compound99089 ), the parent molecule, exhibits an IC50 potency of 3.4 ± 0.5 μM against h12/15-LOX in vitro and an ex vivo IC50 potency of approximately 10 μM in a mouse neuronal cell line, HT-22. Compound99089 displays greater than 30-fold selectivity versus h5-LOX and COX-2, 15-fold versus h15-LOX-2 and 10-fold versus h12-LOX, when tested at 20 μM inhibitor concentration. Steady-state inhibition kinetics reveals that the mode of inhibition of99089 against h12/15-LOX is that of a mixed inhibitor with a K ic of 1.0 ± 0.08 μM and a K iu of 6.0 ± 3.3 μM. These data indicate that99089 and related derivatives may serve as a starting point for the development of anti-stroke therapeutics due to their ability to selectively target h12/15-LOX in vitro and m12/15-LOX ex vivo.