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

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• 作者：Michelle M. Armstrong^a ; Cody J. Freedman^a ; Joo Eun Jung^b ; Yi Zheng^b ; Chakrapani Kalyanaraman^c ; Matthew P. Jacobson^c ; ^&dagger ; ; Anton Simeonov^d ; David J. Maloney^d ; Klaus van Leyen^b ; Ajit Jadhav^d ; Theodore R. Holman^a ; ^{holman@ucsc.edu" class="auth_mail" title="E-mail the corresponding author}
• 关键词：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)-hydroperoxyoctadecadien
• 刊名：Bioorganic & Medicinal Chemistry
• 出版年：2016
• 出版时间：15 March 2016
• 年：2016
• 卷：24
• 期：6
• 页码：1183-1190
• 全文大小：2728 K

文摘

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 K_ic of 1.0 ± 0.08 μM and a K_iu 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.