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Bioorg Med Chem. 2016 Mar 15;24(6):1183-90. doi: 10.1016/j.bmc.2016.01.042. Epub 2016 Jan 21.

A potent and selective inhibitor targeting human and murine 12/15-LOX.

Author information

1
Chemistry and Biochemistry Department, University of California, Santa Cruz, CA 95060, United States.
2
Neuroprotection Research Laboratory, Department of Radiology, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA 02129, United States.
3
Department of Pharmaceutical Chemistry, School of Pharmacy, University of California San Francisco, San Francisco, CA 94143, United States.
4
National Center for Advancing Translational Sciences, National Institutes of Health, 9800 Medical Center Drive, MSC 3370, Bethesda, MD 20892, United States.
5
Chemistry and Biochemistry Department, University of California, Santa Cruz, CA 95060, United States. Electronic address: holman@ucsc.edu.

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 (compound 99089), 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. Compound 99089 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 of 99089 against h12/15-LOX is that of a mixed inhibitor with a Kic of 1.0±0.08 μM and a Kiu of 6.0±3.3 μM. These data indicate that 99089 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.

KEYWORDS:

High-throughput; Human; Inhibitor; Lipoxygenase; Murine; Selective

PMID:
26899595
PMCID:
PMC4778748
DOI:
10.1016/j.bmc.2016.01.042
[Indexed for MEDLINE]
Free PMC Article

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