Novel lipoxygenase inhibitors as neuroprotective reagents

Leyen, Klaus van; Arai, Ken; Jin, Guang; Kenyon, Victor; Gerstner, Bettina; Rosenberg, Paul A.; Holman, Theodore R.; Lo, Eng H.

doi:10.1002/jnr.21543

Cited by 70 publications

(70 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…OPCs were prepared as previously described with some modifications (Ness et al, 2005;van Leyen et al, 2008). Briefly, cerebral cortices from 1-to 2-d-old Sprague Dawley rats were dissected, minced, and digested.…”

Section: Methodsmentioning

confidence: 99%

An Oligovascular Niche: Cerebral Endothelial Cells Promote the Survival and Proliferation of Oligodendrocyte Precursor Cells

Arai

Lo²

2009

J. Neurosci.

220

230

View full text Add to dashboard Cite

We show that cerebral endothelial cells secrete trophic factors that support the survival and proliferation of rat oligodendrocyte precursor cells (OPCs). This OPC-supportive phenomenon was mediated by Akt and Src signaling pathways. Noncytotoxic levels of oxidative stress downregulate trophic factor production and disrupt the ability of cerebral endothelial cells to support OPCs. These data suggest that a novel oligovascular niche may be important for sustaining oligodendrocyte renewal and homeostasis in mammalian brain.

show abstract

Section: Methodsmentioning

confidence: 99%

An Oligovascular Niche: Cerebral Endothelial Cells Promote the Survival and Proliferation of Oligodendrocyte Precursor Cells

Arai

Lo²

2009

J. Neurosci.

220

230

View full text Add to dashboard Cite

show abstract

“…Further studies using 15-LO-1 inhibitors might reveal novel functions for this enzyme in a similar manner as it was described to play a role in cell death in both neurons and oligodendrocytes. 21 In summary, we have used enzyme and cell-based 15-LO-1 assays to successfully identify new potent 15-LO-1 inhibitors for the further development into unique lead series. …”

Section: Resultsmentioning

confidence: 99%

“…21 The importance of 15-LO-1 in several diseases has provoked numerous attempts to identify specific inhibitors, including screening of chemical compound libraries, virtual screening, and identification of natural products. [21][22][23][24][25][26][27][28][29][30][31][32] Especially interesting was the discovery of potent and cell-active 15-LO inhibitors that recently was described by Bristol-Myer-Squibb. 27 As part of our ongoing studies of the human 15-LO-1 enzyme, we have developed an enzyme-and a cell-based assay amenable for high-throughput screening (HTS).…”

mentioning

confidence: 99%

Development of a Fluorescent Intensity Assay Amenable for High-Throughput Screening for Determining 15-Lipoxygenase Activity

et al. 2010

View full text Add to dashboard Cite

15-Lipoxygenase-1 catalyzes the introduction of molecular oxygen into polyunsaturated fatty acids to form a lipid hydroperoxide. The authors have developed an assay for the detection of lipid hydroperoxides formed by human 15-lipoxygenase (15-LO) in enzyme or cellular assays using either a 96-well or a 384-well format. The assays described take advantage of the ability of lipid hydroperoxides to oxidize nonfluorescent diphenyl-1-pyrenylphosphine (DPPP) to a fluorescent phosphine oxide. Oxidation of DPPP yields a fluorescent compound, which is not sensitive to temperature and is stable for more than 2 h. The assay is sensitive toward inhibition and robust with a Z′ value of 0.79 and 0.4 in a 96-and 384-well format, respectively, and thus amenable for high-throughput screening. The utility of DPPP as a marker for 15-lipoxygenase activity was demonstrated with both enzyme-and cell-based assays for the identification of hits and to determine potency by IC 50 determinations. (Journal of Biomolecular Screening 2010:671-679)

show abstract

“…15-LOX-1 is an important mammalian lipoxygenase that plays a role in the biosynthesis of 15-HPETE, 15-HETE, leukotrienes, lipoxins and eoxins. Several studies describe a role for 15-LOX-1 in allergic airway diseases [12][13][14][15], chronic airway inflammation [16], atherosclerosis [17], cancer [18,19] and more recently in various CNS diseases [20][21][22][23][24][25][26][27][28][29] like Alzheimer's and Parkinson's diseases as well as stroke. Therefore, this enzyme gained attention as a potential drug target and several classes of inhibitors have been described.…”

mentioning

confidence: 99%

“…Therefore, this enzyme gained attention as a potential drug target and several classes of inhibitors have been described. [22,[24][25][26][27][30][31][32][33][34][35][36][37][38] Development of activity-based probes for target enzymes start from irreversible mechanismbased enzyme inhibitors. For soybean lipoxygenase, structural analogs of PUFAs have been reported in which the cis-alkenes are replaced by alkynes that proved to be irreversible inactivators of LOX enzymes.…”

mentioning

confidence: 99%

Activity‐Based Probes for 15‐Lipoxygenase‐1

et al. 2016

View full text Add to dashboard Cite

Human 15-lipoxygenase-1 (15-LOX-1) plays an important role in several inflammatory lung diseases such as asthma, COPD and chronic bronchitis as well as in various CNS diseases like Alzheimer's, Parkinson's and stroke. Activity-based probes of 15-LOX-1 are required to explore the role of this enzyme further and to enable drug discovery. In this study, we developed the first 15-LOX-1 activity-based probe as an efficient chemical tool for activity-based labeling of recombinant 15-LOX-1 that also provides 15-LOX-1 dependent labeling in cell lysates and tissue samples. Mimicking the natural substrate of the enzyme, we designed activity-based probes that covalently bind to the active enzyme and include a terminal alkene as chemical reporter for bioorthogonal linkage of a detectable functionality via the oxidative Heck reaction. We believe that the activity-based labeling of 15-LOX-1 will enable the investigation and identification of this enzyme in complex biological samples, which opens completely new opportunities for drug discovery. Keywords15-LOX-1; activity-based probes; Kitz-Wilson plots; enzyme kinetics; irreversible inhibition Activity-based protein profiling (ABPP) has become a powerful method for the analysis of enzyme function and the selectivity of enzyme inhibitors in complex disease models.[1] In ABPP, small molecule substrate analogues, known as activity-based probes, are used to covalently bind to the active site of enzymes depending on their activity. Currently, many research groups use ABPP to investigate various enzyme classes, like cysteine proteases [2][3][4], serine hydrolases [5,6] Development of activity-based probes for target enzymes start from irreversible mechanismbased enzyme inhibitors. For soybean lipoxygenase, structural analogs of PUFAs have been reported in which the cis-alkenes are replaced by alkynes that proved to be irreversible inactivators of LOX enzymes. [39,40] Inactivation is expected to proceed through single electron oxidation of the bis-propargylic carbon resulting in an allene radical that is highly reactive and binds covalently to the enzyme's active site. We aimed to use this type of inhibitor as starting structure to develop activity based probes for LOX enzymes.Mimicking the natural 15-LOX-1 substrate, linoleic acid, inhibitors were designed incorporating a bis-alkyne core structure and their binding properties were investigated.[39,40] After modeling studies, in contrast with the previous inhibitors we shift the position of the bis-alkyne moiety from 9,12 to the 5,8 position due to structural differences in the active sites between the two enzymes ( Figure S9) but also aiming to yield less lipophilic compounds. Next, we developed ABPP probes that include both a bis-alkyne functionality for covalent linkage to the active enzyme and a terminal alkene as chemical reporter for bioorthogonal linkage of a detectable functionality (Figure 1). Application of a terminal alkene as chemical reporter and not the more commonly used terminal alkyne enables straightforward ...

show abstract

Novel lipoxygenase inhibitors as neuroprotective reagents

Cited by 70 publications

References 20 publications

An Oligovascular Niche: Cerebral Endothelial Cells Promote the Survival and Proliferation of Oligodendrocyte Precursor Cells

An Oligovascular Niche: Cerebral Endothelial Cells Promote the Survival and Proliferation of Oligodendrocyte Precursor Cells

Development of a Fluorescent Intensity Assay Amenable for High-Throughput Screening for Determining 15-Lipoxygenase Activity

Activity‐Based Probes for 15‐Lipoxygenase‐1

Contact Info

Product

Resources

About