Morisseau C

General

Full name : Morisseau Christophe

First name : Christophe

Mail : Department of Entomology and Nematology & UCD Comprehensive Cancer Center, University of California, One Shields Avenue, Davis, CA 95616

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Country : USA

Email : chmorisseau@ucdavis.edu

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References (221)

Title : Effects of sEH inhibition on the eicosanoid and cytokine storms in SARS-CoV-2-infected mice - Edin_2024_Faseb.j_38_e23692
Author(s) : Edin ML , Gruzdev A , Graves JP , Lih FB , Morisseau C , Ward JM , Hammock BD , Bosio CM , Zeldin DC
Ref : Faseb j , 38 :e23692 , 2024
Abstract : Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection involves an initial viral infection phase followed by a host-response phase that includes an eicosanoid and cytokine storm, lung inflammation and respiratory failure. While vaccination and early anti-viral therapies are effective in preventing or limiting the pathogenic host response, this latter phase is poorly understood with no highly effective treatment options. Inhibitors of soluble epoxide hydrolase (sEH) increase levels of anti-inflammatory molecules called epoxyeicosatrienoic acids (EETs). This study aimed to investigate the impact of sEH inhibition on the host response to SARS-CoV-2 infection in a mouse model with human angiotensin-converting enzyme 2 (ACE2) expression. Mice were infected with SARS-CoV-2 and treated with either vehicle or the sEH inhibitor 1-trifluoromethoxyphenyl-3-(1-propionylpiperidin-4-yl) urea (TPPU). At day 5 post-infection, SARS-CoV-2 induced weight loss, clinical signs, a cytokine storm, an eicosanoid storm, and severe lung inflammation with ~50% mortality on days 6-8 post-infection. SARS-CoV-2 infection induced lung expression of phospholipase A(2) (PLA(2)), cyclooxygenase (COX) and lipoxygenase (LOX) pathway genes, while suppressing expression of most cytochrome P450 genes. Treatment with the sEH inhibitor TPPU delayed weight loss but did not alter clinical signs, lung cytokine expression or overall survival of infected mice. Interestingly, TPPU treatment significantly reversed the eicosanoid storm and attenuated viral-induced elevation of 39 fatty acids and oxylipins from COX, LOX and P450 pathways, which suggests the effects at the level of PLA(2) activation. The suppression of the eicosanoid storm by TPPU without corresponding changes in lung cytokines, lung inflammation or mortality reveals a surprising dissociation between systemic oxylipin and cytokine signaling pathways during SARS-CoV-2 infection and suggests that the cytokine storm is primarily responsible for morbidity and mortality in this animal model.
ESTHER : Edin_2024_Faseb.j_38_e23692
PubMedSearch : Edin_2024_Faseb.j_38_e23692
PubMedID: 38786655

Title : Design and Synthesis of Dual-Targeting Inhibitors of sEH and HDAC6 for the Treatment of Neuropathic Pain and Lipopolysaccharide-Induced Mortality - Chen_2024_J.Med.Chem__
Author(s) : Chen Y , Sun J , Tong H , Wang J , Cao R , Xu H , Chen L , Morisseau C , Zhang M , Shi Y , Han C , Zhuang J , Jing Y , Liu Z , Hammock BD , Chen G
Ref : Journal of Medicinal Chemistry , : , 2024
Abstract : Epoxyeicosatrienoic acids with anti-inflammatory effects are inactivated by soluble epoxide hydrolase (sEH). Both sEH and histone deacetylase 6 (HDAC6) inhibitors are being developed as neuropathic pain relieving agents. Based on the structural similarity, we designed a new group of compounds with inhibition of both HDAC6 and sEH and obtained compound M9. M9 exhibits selective inhibition of HDAC6 over class I HDACs in cells. M9 shows good microsomal stability, moderate plasma protein binding rate, and oral bioavailability. M9 exhibited a strong analgesic effect in vivo, and its analgesic tolerance was better than gabapentin. M9 improved the survival time of mice treated with lipopolysaccharide (LPS) and reversed the levels of inflammatory factors induced by LPS in mouse plasma. M9 represents the first sEH/HDAC6 dual inhibitors with in vivo antineuropathic pain and anti-inflammation.
ESTHER : Chen_2024_J.Med.Chem__
PubMedSearch : Chen_2024_J.Med.Chem__
PubMedID: 38236416

Title : Inhibition of Soluble Epoxide Hydrolase Does Not Promote or Aggravate Pulmonary Hypertension in Rats - Leuillier_2023_Cells_12_
Author(s) : Leuillier M , Platel V , Tu L , Feugray G , Thuillet R , Groussard D , Messaoudi H , Ottaviani M , Chelgham M , Nicol L , Mulder P , Humbert M , Richard V , Morisseau C , Brunel V , Duflot T , Guignabert C , Bellien J
Ref : Cells , 12 : , 2023
Abstract : Inhibitors of soluble epoxide hydrolase (sEH), which catalyzes the hydrolysis of various natural epoxides to their corresponding diols, present an opportunity for developing oral drugs for a range of human cardiovascular and inflammatory diseases, including, among others, diabetes and neuropathic pain. However, some evidence suggests that their administration may precipitate the development of pulmonary hypertension (PH). We thus evaluated the impact of chronic oral administration of the sEH inhibitor TPPU (N-[1-(1-Oxopropyl)-4-piperidinyl]-N'-[4-(trifluoromethoxy)phenyl]-urea) on hemodynamics, pulmonary vascular reactivity, and remodeling, as well as on right ventricular (RV) dimension and function at baseline and in the Sugen (SU5416) + hypoxia (SuHx) rat model of severe PH. Treatment with TPPU started 5 weeks after SU5416 injection for 3 weeks. No differences regarding the increase in pulmonary vascular resistance, remodeling, and inflammation, nor the abolishment of phenylephrine-induced pulmonary artery constriction, were noted in SuHx rats. In addition, TPPU did not modify the development of RV dysfunction, hypertrophy, and fibrosis in SuHx rats. Similarly, none of these parameters were affected by TPPU in normoxic rats. Complementary in vitro data demonstrated that TPPU reduced the proliferation of cultured human pulmonary artery-smooth muscle cells (PA-SMCs). This study demonstrates that inhibition of sEH does not induce nor aggravate the development of PH and RV dysfunction in SuHx rats. In contrast, a potential beneficial effect against pulmonary artery remodeling in humans is suggested.
ESTHER : Leuillier_2023_Cells_12_
PubMedSearch : Leuillier_2023_Cells_12_
PubMedID: 36831332

Title : Direct targeting of sEH with alisol B alleviated the apoptosis, inflammation, and oxidative stress in cisplatin-induced acute kidney injury - Zhang_2023_Int.J.Biol.Sci_19_294
Author(s) : Zhang J , Luan ZL , Huo XK , Zhang M , Morisseau C , Sun CP , Hammock BD , Ma XC
Ref : Int J Biol Sci , 19 :294 , 2023
Abstract : Acute kidney injury (AKI) is a pathological condition characterized by a rapid decrease in glomerular filtration rate and nitrogenous waste accumulation during hemodynamic regulation. Alisol B, from Alisma orientale, displays anti-tumor, anti-complement, and anti-inflammatory effects. However, its effect and action mechanism on AKI is still unclear. Herein, alisol B significantly attenuated cisplatin (Cis)-induced renal tubular apoptosis through decreasing expressions levels of cleaved-caspase 3 and cleaved-PARP and the ratio of Bax/Bcl-2 depended on the p53 pathway. Alisol B also alleviated Cis-induced inflammatory response (e.g. the increase of ICAM-1, MCP-1, COX-2, iNOS, IL-6, and TNF-alpha) and oxidative stress (e.g. the decrease of SOD and GSH, the decrease of HO-1, GCLC, GCLM, and NQO-1) through the NF-kappaB and Nrf2 pathways. In a target fishing experiment, alisol B bound to soluble epoxide hydrolase (sEH) as a direct cellular target through the hydrogen bond with Gln384, which was further supported by inhibition kinetics and surface plasmon resonance (equilibrium dissociation constant, K (D) = 1.32 microM). Notably, alisol B enhanced levels of epoxyeicosatrienoic acids and decreased levels of dihydroxyeicosatrienoic acids, indicating that alisol B reduced the sEH activity in vivo. In addition, sEH genetic deletion alleviated Cis-induced AKI and abolished the protective effect of alisol B in Cis-induced AKI as well. These findings indicated that alisol B targeted sEH to alleviate Cis-induced AKI via GSK3beta-mediated p53, NF-kappaB, and Nrf2 signaling pathways and could be used as a potential therapeutic agent in the treatment of AKI.
ESTHER : Zhang_2023_Int.J.Biol.Sci_19_294
PubMedSearch : Zhang_2023_Int.J.Biol.Sci_19_294
PubMedID: 36594097

Title : Quantification of soluble epoxide hydrolase inhibitors in experimental and clinical samples using the nanobody-based ELISA - Yang_2023_J.Pharm.Anal_13_1013
Author(s) : Yang H , Qi M , He Q , Hwang SH , Yang J , McCoy M , Morisseau C , Zhao S , Hammock BD
Ref : J Pharm Anal , 13 :1013 , 2023
Abstract : To ensure proper dosage of a drug, analytical quantification of it in biofluid is necessary. Liquid chromatography mass spectrometry (LC-MS) is the conventional method of choice as it permits accurate identification and quantification. However, it requires expensive instrumentation and is not appropriate for bedside use. Using soluble epoxide hydrolase (sEH) inhibitors (EC5026 and TPPU) as examples, we report development of a nanobody-based enzyme-linked immunosorbent assay (ELISA) for such small molecules and its use to accurately quantify the drug chemicals in human samples. Under optimized conditions, two nanobody-based ELISAs were successfully established for EC5026 and TPPU with low limits of detection of 0.085 ng/mL and 0.31 ng/mL, respectively, and two order of magnitude linear ranges with high precision and accuracy. The assay was designed to detect parent and two biologically active metabolites in the investigation of a new drug candidate EC5026. In addition, the ELISAs displayed excellent correlation with LC-MS analysis and evaluation of inhibitory potency. The results indicate that nanobody-based ELISA methods can efficiently analyze drug like compounds. These methods could be easily implemented by the bedside, in the field in remote areas or in veterinary practice. This work illustrates that nanobody based assays offer alternative and supplementary analytical tools to mass spectrometry for monitoring small molecule medicines during clinical development and therapy. Attributes of nanobody based pharmaceutical assays are discussed.
ESTHER : Yang_2023_J.Pharm.Anal_13_1013
PubMedSearch : Yang_2023_J.Pharm.Anal_13_1013
PubMedID: 37842656

Title : Soluble epoxide hydrolase-targeting PROTAC activates AMPK and inhibits endoplasmic reticulum stress - Peyman_2023_Biomed.Pharmacother_168_115667
Author(s) : Peyman M , Barroso E , Turcu AL , Estrany F, Jr. , Smith D , Jurado-Aguilar J , Rada P , Morisseau C , Hammock BD , Valverde A M , Palomer X , Galdeano C , Vazquez S , Vazquez-Carrera M
Ref : Biomed Pharmacother , 168 :115667 , 2023
Abstract : Soluble epoxide hydrolase (sEH) is a drug target with the potential for therapeutic utility in the areas of inflammation, neurodegenerative disease, chronic pain, and diabetes, among others. Proteolysis-targeting chimeras (PROTACs) molecules offer new opportunities for targeting sEH, due to its capacity to induce its degradation. Here, we describe that the new ALT-PG2, a PROTAC that degrades sEH protein in the human hepatic Huh-7 cell line, in isolated mouse primary hepatocytes, and in the liver of mice. Remarkably, sEH degradation caused by ALT-PG2 was accompanied by an increase in the phosphorylated levels of AMP-activated protein kinase (AMPK), while phosphorylated extracellular-signal-regulated kinase 1/2 (ERK1/2) was reduced. Consistent with the key role of these kinases on endoplasmic reticulum (ER) stress, ALT-PG2 attenuated the levels of ER stress and inflammatory markers. Overall, the findings of this study indicate that targeting sEH with degraders is a promising pharmacological strategy to promote AMPK activation and to reduce ER stress and inflammation.
ESTHER : Peyman_2023_Biomed.Pharmacother_168_115667
PubMedSearch : Peyman_2023_Biomed.Pharmacother_168_115667
PubMedID: 37826940
Gene_locus related to this paper: human-EPHX2

Title : PROTAC-Mediated Selective Degradation of Cytosolic Soluble Epoxide Hydrolase Enhances ER Stress Reduction - Wang_2023_ACS.Chem.Biol__
Author(s) : Wang Y , Morisseau C , Takamura A , Wan D , Li D , Sidoli S , Yang J , Wolan DW , Hammock BD , Kitamura S
Ref : ACS Chemical Biology , : , 2023
Abstract : Soluble epoxide hydrolase (sEH) is a bifunctional enzyme responsible for lipid metabolism and is a promising drug target. Here, we report the first-in-class PROTAC small-molecule degraders of sEH. Our optimized PROTAC selectively targets the degradation of cytosolic but not peroxisomal sEH, resulting in exquisite spatiotemporal control. Remarkably, our sEH PROTAC molecule has higher potency in cellular assays compared to the parent sEH inhibitor as measured by the significantly reduced ER stress. Interestingly, our mechanistic data indicate that our PROTAC directs the degradation of cytosolic sEH via the lysosome, not through the proteasome. The molecules presented here are useful chemical probes to study the biology of sEH with the potential for therapeutic development. Broadly, our results represent a proof of concept for the superior cellular potency of sEH degradation over sEH enzymatic inhibition, as well as subcellular compartment-selective modulation of a protein by PROTACs.
ESTHER : Wang_2023_ACS.Chem.Biol__
PubMedSearch : Wang_2023_ACS.Chem.Biol__
PubMedID: 36947831
Gene_locus related to this paper: human-EPHX2

Title : Fluorine and chlorine substituted adamantyl-urea as molecular tools for inhibition of human soluble epoxide hydrolase with picomolar efficacy - Burmistrov_2023_J.Enzyme.Inhib.Med.Chem_38_2274797
Author(s) : Burmistrov VV , Morisseau C , Danilov DV , Gladkikh BP , D'Yachenko V S , Zefirov NA , Zefirova ON , Butov GM , Hammock BD
Ref : J Enzyme Inhib Med Chem , 38 :2274797 , 2023
Abstract : Series of 1,3-disubstituted ureas and diadamantyl disubstituted diureas with fluorinated and chlorinated adamantane residues were shown to inhibit human soluble epoxide hydrolase (sEH) with inhibition potency ranging from 40 pM to 9.2 nM. The measured IC(50) values for some molecules were below the accuracy limit of the existing in vitro assays. Such an increase in activity was achieved by minimal structural modifications to the molecules of known inhibitors, including 4-[trans-4-(1-adamantylcarbamoylamino)cyclohexyl]oxybenzoic acid. For the chlorinated homologue of the latter the sharp jump in inhibitory activity can be (according to molecular dynamics data) the result of interactions - Cl-Pi interaction. Considering the extremely high inhibitory activity, acceptable solubility and partial blockage of metabolically sensitive centres in their structures, some compounds are of interest for further in vivo biotesting.
ESTHER : Burmistrov_2023_J.Enzyme.Inhib.Med.Chem_38_2274797
PubMedSearch : Burmistrov_2023_J.Enzyme.Inhib.Med.Chem_38_2274797
PubMedID: 37975322

Title : Neuroprotective effect of herbal extracts inhibiting soluble epoxide hydrolase (sEH) and cyclooxygenase (COX) against chemotherapy-induced cognitive impairment in mice - Kulkarni_2023_Biochem.Biophys.Res.Commun_667_64
Author(s) : Kulkarni R , Mehta R , Goswami SK , Hammock BD , Morisseau C , Hwang SH , Mallappa O , Azeemuddin MM , Rafiq M , S NM
Ref : Biochemical & Biophysical Research Communications , 667 :64 , 2023
Abstract : Chemotherapy-induced cognitive impairment (CICI) is a novel clinical condition characterized by memory, learning, and motor function deficits. Oxidative stress and inflammation are potential factors contributing to chemotherapy's adverse effects on the brain. Inhibition of soluble epoxide hydrolase (sEH) has been proven effective in neuroinflammation and reversal of memory impairment. The research aims to evaluate the memory protective effect of sEH inhibitor and dual inhibitor of sEH and COX and compare its impact with herbal extracts with known nootropic activity in an animal model of CICI. In vitro sEH, the inhibitory activity of hydroalcoholic extracts of Sizygium aromaticum, Nigella sativa, and Mesua ferrea was tested on murine and human sEH enzyme as per the protocol, and IC(50) was determined. Cyclophosphamide (50 mg/kg), methotrexate (5 mg/kg), and fluorouracil (5 mg/kg) combination (CMF) were administered intraperitoneally to induce CICI. The known herbal sEH inhibitor, Lepidium meyenii and the dual inhibitor of COX and sEH (PTUPB) were tested for their protective effect in the CICI model. The herbal formulation with known nootropic activity viz Bacopa monnieri and commercial formulation (Mentat) were also used to compare the efficacy in the CICI model. Behavioral parameter such as cognitive function was assessed by Morris Water Maze besides investigating oxidative stress (GSH and LPO) and inflammatory (TNFalpha, IL-6, BDNF and COX-2) markers in the brain. CMF-induced CICI, which was associated with increased oxidative stress and inflammation in the brain. However, treatment with PTUPB or herbal extracts inhibiting sEH preserved spatial memory via ameliorating oxidative stress and inflammation. S. aromaticum and N. sativa inhibited COX2, but M. Ferrea did not affect COX2 activity. Lepidium meyenii was the least effective, and mentat showed superior activity over Bacopa monnieri in preserving memory. Compared to untreated animals, the mice treated with PTUPB or hydroalcoholic extracts showed a discernible improvement in cognitive function in CICI.
ESTHER : Kulkarni_2023_Biochem.Biophys.Res.Commun_667_64
PubMedSearch : Kulkarni_2023_Biochem.Biophys.Res.Commun_667_64
PubMedID: 37209564

Title : CRISPR\/Cas9-mediated inactivation of the phosphatase activity of soluble epoxide hydrolase prevents obesity and cardiac ischemic injury - Leuillier_2023_J.Adv.Res_43_163
Author(s) : Leuillier M , Duflot T , Menoret S , Messaoudi H , Djerada Z , Groussard D , Denis RGP , Chevalier L , Karoui A , Panthu B , Thiebaut PA , Schmitz-Afonso I , Nobis S , Campart C , Henry T , Sautreuil C , Luquet SH , Beseme O , Feliu C , Peyret H , Nicol L , Henry JP , Renet S , Mulder P , Wan D , Tesson L , Heslan JM , Duche A , Jacques S , Ziegler F , Brunel V , Rautureau GJP , Monteil C , do Rego JL , do Rego JC , Afonso C , Hammock B , Madec AM , Pinet F , Richard V , Anegon I , Guignabert C , Morisseau C , Bellien J
Ref : J Adv Res , 43 :163 , 2023
Abstract : INTRODUCTION: Although the physiological role of the C-terminal hydrolase domain of the soluble epoxide hydrolase (sEH-H) is well investigated, the function of its N-terminal phosphatase activity (sEH-P) remains unknown. OBJECTIVES: This study aimed to assess in vivo the physiological role of sEH-P. METHODS: CRISPR/Cas9 was used to generate a novel knock-in (KI) rat line lacking the sEH-P activity. RESULTS: The sEH-P KI rats has a decreased metabolism of lysophosphatidic acids to monoacyglycerols. KI rats grew almost normally but with less weight and fat mass gain while insulin sensitivity was increased compared to wild-type rats. This lean phenotype was more marked in males than in female KI rats and mainly due to decreased food consumption and enhanced energy expenditure. In fact, sEH-P KI rats had an increased lipolysis allowing to supply fatty acids as fuel to potentiate brown adipose thermogenesis under resting condition and upon cold exposure. The potentiation of thermogenesis was abolished when blocking PPARgamma, a nuclear receptor activated by intracellular lysophosphatidic acids, but also when inhibiting simultaneously sEH-H, showing a functional interaction between the two domains. Furthermore, sEH-P KI rats fed a high-fat diet did not gain as much weight as the wild-type rats, did not have increased fat mass and did not develop insulin resistance or hepatic steatosis. In addition, sEH-P KI rats exhibited enhanced basal cardiac mitochondrial activity associated with an enhanced left ventricular contractility and were protected against cardiac ischemia-reperfusion injury. CONCLUSION: Our study reveals that sEH-P is a key player in energy and fat metabolism and contributes together with sEH-H to the regulation of cardiometabolic homeostasis. The development of pharmacological inhibitors of sEH-P appears of crucial importance to evaluate the interest of this promising therapeutic strategy in the management of obesity and cardiac ischemic complications.
ESTHER : Leuillier_2023_J.Adv.Res_43_163
PubMedSearch : Leuillier_2023_J.Adv.Res_43_163
PubMedID: 36585106
Gene_locus related to this paper: human-EPHX2

Title : Structure-Directed Discovery of Potent Soluble Epoxide Hydrolase Inhibitors for the Treatment of Inflammatory Diseases - Chen_2023_J.Med.Chem__
Author(s) : Chen Y , Chen L , Xu H , Cao R , Morisseau C , Zhang M , Shi Y , Hammock BD , Wang J , Zhuang J , Liu Z , Chen G
Ref : Journal of Medicinal Chemistry , : , 2023
Abstract : Soluble epoxide hydrolase (sEH) has been identified as an attractive target for anti-inflammatory drug design in recent years. Picomolar level compound G1 against sEH was obtained by introducing the hydrophilic group homopiperazine and hydrophobic fragment propionyl onto the structure of lead compound A. G1 showed good microsomal stability, a moderate plasma protein binding rate, and good oral bioavailability and was well tolerated in rats. G1 has significant analgesic effects on CFA-induced AIA mice, ameliorated the pancreatic injury in acute pancreatitis induced by l-arginine, reversed pancreatic injury, edema, and neutrophil infiltration, and increased the survival time of C57BL/6 mice in a lipopolysaccharide (LPS)-induced sepsis model. Moreover the expression levels of sEH, COX-2, NOS-2, vascular cell adhesion molecule (VCAM), IL-6, MCP-5, and tumor necrosis factor alpha (TNF-alpha) were measured by Western blot or enzyme-linked immunosorbent assay (ELISA), with varying degrees of decrease. These results suggested that G1 is a drug candidate worthy of further evaluation for the treatment of inflammation-induced diseases such as arthritis, acute pancreatitis, and sepsis.
ESTHER : Chen_2023_J.Med.Chem__
PubMedSearch : Chen_2023_J.Med.Chem__
PubMedID: 36689364

Title : Aflatoxin B(1) Increases Soluble Epoxide Hydrolase in the Brain and Induces Neuroinflammation and Dopaminergic Neurotoxicity - Wang_2023_Int.J.Mol.Sci_24_9938
Author(s) : Wang W , Wang Y , Wagner KM , Lee RD , Hwang SH , Morisseau C , Wulff H , Hammock BD
Ref : Int J Mol Sci , 24 :9938 , 2023
Abstract : Parkinson's disease (PD) is an increasingly common neurodegenerative movement disorder with contributing factors that are still largely unexplored and currently no effective intervention strategy. Epidemiological and pre-clinical studies support the close association between environmental toxicant exposure and PD incidence. Aflatoxin B(1) (AFB(1)), a hazardous mycotoxin commonly present in food and environment, is alarmingly high in many areas of the world. Previous evidence suggests that chronic exposure to AFB(1) leads to neurological disorders as well as cancer. However, whether and how aflatoxin B(1) contributes to the pathogenesis of PD is poorly understood. Here, oral exposure to AFB(1) is shown to induce neuroinflammation, trigger the alpha-synuclein pathology, and cause dopaminergic neurotoxicity. This was accompanied by the increased expression and enzymatic activity of soluble epoxide hydrolase (sEH) in the mouse brain. Importantly, genetic deletion or pharmacological inhibition of sEH alleviated the AFB(1)-induced neuroinflammation by reducing microglia activation and suppressing pro-inflammatory factors in the brain. Furthermore, blocking the action of sEH attenuated dopaminergic neuron dysfunction caused by AFB(1) in vivo and in vitro. Together, our findings suggest a contributing role of AFB(1) to PD etiology and highlight sEH as a potential pharmacological target for alleviating PD-related neuronal disorders caused by AFB(1) exposure.
ESTHER : Wang_2023_Int.J.Mol.Sci_24_9938
PubMedSearch : Wang_2023_Int.J.Mol.Sci_24_9938
PubMedID: 37373086

Title : Determinants of Meal-Induced Changes in Circulating FFA Epoxides, Diols, and Diol-to-Epoxide Ratios as Indices of Soluble Epoxide Hydrolase Activity - Oh_2023_Int.J.Mol.Sci_24_
Author(s) : Oh YT , Yang J , Morisseau C , He Q , Hammock B , Youn JH , Stefanovski D
Ref : Int J Mol Sci , 24 : , 2023
Abstract : Soluble epoxide hydrolase (sEH) is an important enzyme for metabolic and cardiovascular health. sEH converts FFA epoxides (EpFAs), many of which are regulators of various cellular processes, to biologically less active diols. In human studies, diol (sEH product) to EpFA (sEH substrate) ratios in plasma or serum have been used as indices of sEH activity. We previously showed these ratios profoundly decreased in rats during acute feeding, possibly reflecting decreases in tissue sEH activities. The present study was designed to test which tissue(s) these measurements in the blood represent and if factors other than sEH activity, such as renal excretion or dietary intake of EpFAs and diols, significantly alter plasma EpFAs, diols, and/or their ratios. The results show that postprandial changes in EpFAs and diols and their ratios in plasma were very similar to those observed in the liver but not in other tissues, suggesting that the liver is largely responsible for these changes in plasma levels. EpFAs and diols were excreted into the urine, but their levels were not significantly altered by feeding, suggesting that renal excretion of EpFAs and diols may not play a major role in postprandial changes in circulating EpFAs, diols, or their ratios. Diet intake had significant impacts on circulating EpFA and diol levels but not on diol-to-EpFA (D-to-E) ratios, suggesting that these ratios, reflecting sEH activities, may not be significantly affected by the availability of sEH substrates (i.e., EpFAs). In conclusion, changes in FFA D-to-E ratios in plasma may reflect those in the liver, which may in turn represent sEH activities in the liver, and they may not be significantly affected by renal excretion or the dietary intake of EpFAs and diols.
ESTHER : Oh_2023_Int.J.Mol.Sci_24_
PubMedSearch : Oh_2023_Int.J.Mol.Sci_24_
PubMedID: 37445935 || 38139180

Title : Screening and Biological Evaluation of Soluble Epoxide Hydrolase Inhibitors: Assessing the Role of Hydrophobicity in the Pharmacophore-Guided Search of Novel Hits - Vazquez_2023_J.Chem.Inf.Model__
Author(s) : Vazquez J , Ginex T , Herrero A , Morisseau C , Hammock BD , Luque FJ
Ref : J Chem Inf Model , : , 2023
Abstract : The human soluble epoxide hydrolase (sEH) is a bifunctional enzyme that modulates the levels of regulatory epoxy lipids. The hydrolase activity is carried out by a catalytic triad located at the center of a wide L-shaped binding site, which contains two hydrophobic subpockets at both sides. On the basis of these structural features, it can be assumed that desolvation is a major factor in determining the maximal achievable affinity that can be attained for this pocket. Accordingly, hydrophobic descriptors may be better suited to the search of novel hits targeting this enzyme. This study examines the suitability of quantum mechanically derived hydrophobic descriptors in the discovery of novel sEH inhibitors. To this end, three-dimensional quantitative structure-activity relationship (3D-QSAR) pharmacophores were generated by combining electrostatic and steric or alternatively hydrophobic and hydrogen-bond parameters in conjunction with a tailored list of 76 known sEH inhibitors. The pharmacophore models were then validated by using two external sets chosen (i) to rank the potency of four distinct series of compounds and (ii) to discriminate actives from decoys, using in both cases datasets taken from the literature. Finally, a prospective study was performed including a virtual screening of two chemical libraries to identify new potential hits, which were subsequently experimentally tested for their inhibitory activity on human, rat, and mouse sEH. The use of hydrophobic-based descriptors led to the identification of six compounds as inhibitors of the human enzyme with IC(50) < 20 nM, including two with IC(50) values of 0.4 and 0.7 nM. The results support the use of hydrophobic descriptors as a valuable tool in the search of novel scaffolds that encode a proper hydrophilic/hydrophobic distribution complementary to the target's binding site.
ESTHER : Vazquez_2023_J.Chem.Inf.Model__
PubMedSearch : Vazquez_2023_J.Chem.Inf.Model__
PubMedID: 37141492

Title : Mix-and-Read Nanobody-Based Sandwich Homogeneous Split-Luciferase Assay for the Rapid Detection of Human Soluble Epoxide Hydrolase - He_2023_Anal.Chem__
Author(s) : He Q , McCoy MR , Yang H , Lin M , Cui X , Zhao S , Morisseau C , Li D , Hammock BD
Ref : Analytical Chemistry , : , 2023
Abstract : The soluble epoxide hydrolase (sEH) is possibly both a marker for and target of numerous diseases. Herein, we describe a homogeneous mix-and-read assay for the detection of human sEH based on using split-luciferase detection coupled with anti-sEH nanobodies. Selective anti-sEH nanobodies were individually fused with NanoLuc Binary Technology (NanoBiT), which consists of a large and small portion of NanoLuc (LgBiT and SmBiT, respectively). Different orientations of the LgBiT and SmBiT-nanobody fusions were expressed and investigated for their ability to reform the active NanoLuc in the presence of the sEH. After optimization, the linear range of the assay could reach 3 orders of magnitude with a limit of detection (LOD) of 1.4 ng/mL. The assay has a high sensitivity to human sEH and reached a similar detection limit to our previously reported conventional nanobody-based ELISA. The procedure of the assay was faster (30 min total) and easy to operate, providing a more flexible and simple way to monitor human sEH levels in biological samples. In general, the immunoassay proposed here offers a more efficient detection and quantification approach that can be easily adapted to numerous macromolecules.
ESTHER : He_2023_Anal.Chem__
PubMedSearch : He_2023_Anal.Chem__
PubMedID: 36972550

Title : The Generation of a Nanobody-Based ELISA for Human Microsomal Epoxide Hydrolase - He_2023_Int.J.Mol.Sci_24_
Author(s) : He Q , McCoy MR , Qi M , Morisseau C , Yang H , Xu C , Shey R , Goodman MC , Zhao S , Hammock BD
Ref : Int J Mol Sci , 24 : , 2023
Abstract : A microsomal epoxide hydrolase (mEH) metabolizes in vivo in both xenobiotic and endogenous epoxides associated with signaling function. Findings in patients suggest that mEH might be a biomarker for several diseases, including metastatic cancer and viral hepatitis. To easily quantify mEH, nanobodies specific to the human mEH were isolated from a phage library of llama VHHs. Four unique clones were obtained and used for developing ELISAs. Three formats of double antibody sandwich assays were investigated using different detection strategies. Using PolyHRP, the signal was strongly amplified, yielding a 22-fold lower LOD (12 pg mL(-1)) than the 'conventional'. To further validate the performance of the immunoassays, human tissue samples were analyzed by nanobody-based ELISAs and compared to the enzyme activities (R(2) > 0.95). The results demonstrate that these nanobodies are powerful tools for the quantification of human mEH and could eventually result in a bedside assay.
ESTHER : He_2023_Int.J.Mol.Sci_24_
PubMedSearch : He_2023_Int.J.Mol.Sci_24_
PubMedID: 37834144

Title : Regulatory lipid vicinal diols counteract the biological activity of epoxy fatty acids and can act as biomarkers and mechanisms for disease progression - McReynolds_2023_Pharmacol.Ther__108454
Author(s) : McReynolds C , Hammock B , Morisseau C
Ref : Pharmacol Ther , :108454 , 2023
Abstract : Polyunsaturated fatty acids (PUFAs) are essential fatty acids required for human health and are obtained primarily from food or synthesized in the body by highly regulated processes. The metabolites of these lipids, formed largely through the action of cyclooxygenase, lipoxygenase, or cytochrome P450 (CYP450) enzymes, are responsible for multiple biological functions including inflammation, tissue repair, cell proliferation, blood vessel permeability, and immune cell behavior. The role of these regulatory lipids in disease has been well studied since their discovery as druggable targets; however, the metabolites generated downstream of these pathways have only recently gained attention for regulating biology. Specifically, the biological activity of lipid vicinal diols formed from the metabolism of CYP450-generated epoxy fatty acids (EpFA) by epoxide hydrolases were previously thought to have little biological activity but increasingly are recognized as promoting inflammation and brown fat adipogenesis, and exciting neurons through the regulation of ion channel activity at low concentrations. These metabolites also appear to balance the action of the EpFA precursor. For example, EpFA demonstrate the ability to resolve inflammation and reduce pain, while some lipid diols, through opposing mechanisms, promote inflammation and pain. This review describes recent studies that highlight the role of regulatory lipids, focusing on the balance between EpFA and their diol metabolites in promoting or resolving disease.
ESTHER : McReynolds_2023_Pharmacol.Ther__108454
PubMedSearch : McReynolds_2023_Pharmacol.Ther__108454
PubMedID: 37268114

Title : Macrophage Inactivation by Small Molecule Wedelolactone via Targeting sEH for the Treatment of LPS-Induced Acute Lung Injury - Zhang_2023_ACS.Cent.Sci_9_440
Author(s) : Zhang J , Zhang M , Huo XK , Ning J , Yu ZL , Morisseau C , Sun CP , Hammock BD , Ma XC
Ref : ACS Cent Sci , 9 :440 , 2023
Abstract : Soluble epoxide hydrolase (sEH) plays a critical role in inflammation by modulating levels of epoxyeicosatrienoic acids (EETs) and other epoxy fatty acids (EpFAs). Here, we investigate the possible role of sEH in lipopolysaccharide (LPS)-mediated macrophage activation and acute lung injury (ALI). In this study, we found that a small molecule, wedelolactone (WED), targeted sEH and led to macrophage inactivation. Through the molecular interaction with amino acids Phe362 and Gln384, WED suppressed sEH activity to enhance levels of EETs, thus attenuating inflammation and oxidative stress by regulating glycogen synthase kinase 3beta (GSK3beta)-mediated nuclear factor-kappa B (NF-kappaB) and nuclear factor E2-related factor 2 (Nrf2) pathways in vitro. In an LPS-stimulated ALI animal model, pharmacological sEH inhibition by WED or sEH knockout (KO) alleviated pulmonary damage, such as the increase in the alveolar wall thickness and collapse. Additionally, WED or sEH genetic KO both suppressed macrophage activation and attenuated inflammation and oxidative stress in vivo. These findings provided the broader prospects for ALI treatment by targeting sEH to alleviate inflammation and oxidative stress and suggested WED as a natural lead candidate for the development of novel synthetic sEH inhibitors.
ESTHER : Zhang_2023_ACS.Cent.Sci_9_440
PubMedSearch : Zhang_2023_ACS.Cent.Sci_9_440
PubMedID: 36968547

Title : Soluble Epoxide Hydrolase Contributes to Cell Senescence and ER Stress in Aging Mice Colon - Wang_2023_Int.J.Mol.Sci_24_4570
Author(s) : Wang W , Wagner KM , Wang Y , Singh N , Yang J , He Q , Morisseau C , Hammock BD
Ref : Int J Mol Sci , 24 :4570 , 2023
Abstract : Aging, which is characterized by enhanced cell senescence and functional decline of tissues, is a major risk factor for many chronic diseases. Accumulating evidence shows that age-related dysfunction in the colon leads to disorders in multiple organs and systemic inflammation. However, the detailed pathological mechanisms and endogenous regulators underlying colon aging are still largely unknown. Here, we report that the expression and activity of the soluble epoxide hydrolase (sEH) enzyme are increased in the colon of aged mice. Importantly, genetic knockout of sEH attenuated the age-related upregulation of senescent markers p21, p16, Tp53, and beta-galactosidase in the colon. Moreover, sEH deficiency alleviated aging-associated endoplasmic reticulum (ER) stress in the colon by reducing both the upstream regulators Perk and Ire1 as well as the downstream pro-apoptotic effectors Chop and Gadd34. Furthermore, treatment with sEH-derived linoleic acid metabolites, dihydroxy-octadecenoic acids (DiHOMEs), decreased cell viability and increased ER stress in human colon CCD-18Co cells in vitro. Together, these results support that the sEH is a key regulator of the aging colon, which highlights its potential application as a therapeutic target for reducing or treating age-related diseases in the colon.
ESTHER : Wang_2023_Int.J.Mol.Sci_24_4570
PubMedSearch : Wang_2023_Int.J.Mol.Sci_24_4570
PubMedID: 36901999

Title : Genetic deletion or pharmacological inhibition of soluble epoxide hydrolase attenuated particulate matter 2.5 exposure mediated lung injury - Zhang_2023_J.Hazard.Mater_458_131890
Author(s) : Zhang J , Zhang WH , Morisseau C , Zhang M , Dong HJ , Zhu QM , Huo XK , Sun CP , Hammock BD , Ma XC
Ref : J Hazard Mater , 458 :131890 , 2023
Abstract : Air pollution represented by particulate matter 2.5 (PM2.5) is closely related to diseases of the respiratory system. Although the understanding of its mechanism is limited, pulmonary inflammation is closely correlated with PM2.5-mediated lung injury. Soluble epoxide hydrolase (sEH) and epoxy fatty acids play a vital role in the inflammation. Herein, we attempted to use the metabolomics of oxidized lipids for analyzing the relationship of oxylipins with lung injury in a PM2.5-mediated mouse model, and found that the cytochrome P450 oxidases/sEH mediated metabolic pathway was involved in lung injury. Furthermore, the sEH overexpression was revealed in lung injury mice. Interestingly, sEH genetic deletion or the selective sEH inhibitor TPPU increased levels of epoxyeicosatrienoic acids (EETs) in lung injury mice, and inactivated pulmonary macrophages based on the MAPK/NF-kappaB pathway, resulting in protection against PM2.5-mediated lung injury. Additionally, a natural sEH inhibitor luteolin from Inula japonica displayed a pulmonary protective effect towards lung injury mediated by PM2.5 as well. Our results are consistent with the sEH message and protein being both a marker and mechanism for PM2.5-induced inflammation, which suggest its potential as a pharmaceutical target for treating diseases of the respiratory system.
ESTHER : Zhang_2023_J.Hazard.Mater_458_131890
PubMedSearch : Zhang_2023_J.Hazard.Mater_458_131890
PubMedID: 37406527

Title : Adamantyl-ureas with pyrazoles substituted by fluoroalkanes as soluble epoxide hydrolase inhibitors - Burmistrov_2023_J.Fluor.Chem_266_
Author(s) : Burmistrov VV , Morisseau C , Shkineva TK , Danilov DV , Gladkikh B , Butov GM , Fayzullin RR , Dutova TY , Hammock BD , Dalinger IL
Ref : J Fluor Chem , 266 : , 2023
Abstract : A series of soluble epoxide hydrolase (sEH) inhibitors containing halogenated pyrazoles was developed. Inhibition potency of the obtained compounds ranges from 0.8 to 27.5 nM. 1-Adamantyl-3-[(4,5-dichloro-1-methyl-1-pyrazol-3-yl)methyl]urea (3f, IC(50) = 0.8 nM) and 1-[(Adamantan-1-yl)methyl]-3-[(4,5-dichloro-1-methyl-1-pyrazol-3-yl)methyl]urea (4f, IC(50) = 1.2 nM) were found to be the most potent sEH inhibitors within the described series.
ESTHER : Burmistrov_2023_J.Fluor.Chem_266_
PubMedSearch : Burmistrov_2023_J.Fluor.Chem_266_
PubMedID: 37638129

Title : Soluble Epoxide Hydrolase Is Associated with Postprandial Anxiety Decrease in Healthy Adult Women - Nguyen_2022_Int.J.Mol.Sci_23_
Author(s) : Nguyen N , Morisseau C , Li D , Yang J , Lam E , Woodside DB , Hammock BD , Shih PB
Ref : Int J Mol Sci , 23 : , 2022
Abstract : The metabolism of bioactive oxylipins by soluble epoxide hydrolase (sEH) plays an important role in inflammation, and sEH may be a risk modifier in various human diseases and disorders. The relationships that sEH has with the risk factors of these diseases remain elusive. Herein, sEH protein expression and activity in white blood cells were characterized before and after a high-fat meal in healthy women (HW) and women with anorexia nervosa (AN). sEH expression and sEH activity were significantly correlated and increased in both groups two hours after consumption of the study meal. Fasting sEH expression and activity were positively associated with body mass index (BMI) in both groups, while an inverse association with age was found in AN only (p value < 0.05). sEH was not associated with anxiety or depression in either group at the fasting timepoint. While the anxiety score decreased after eating in both groups, a higher fasting sEH was associated with a lower postprandial anxiety decrease in HW (p value < 0.05). sEH characterization using direct measurements verified the relationship between the protein expression and in vivo activity of this important oxylipin modulator, while a well-controlled food challenge study design using HW and a clinical control group of women with disordered eating elucidated sEH's role in the health of adult women.
ESTHER : Nguyen_2022_Int.J.Mol.Sci_23_
PubMedSearch : Nguyen_2022_Int.J.Mol.Sci_23_
PubMedID: 36233100

Title : Kurarinone alleviated Parkinson's disease via stabilization of epoxyeicosatrienoic acids in animal model - Sun_2022_Proc.Natl.Acad.Sci.U.S.A_119_
Author(s) : Sun CP , Zhou JJ , Yu ZL , Huo XK , Zhang J , Morisseau C , Hammock BD , Ma XC
Ref : Proc Natl Acad Sci U S A , 119 : , 2022
Abstract : Parkinson's disease (PD) is one of the most common neurodegenerative disorders and is characterized by loss of dopaminergic neurons in the substantia nigra (SN), causing bradykinesia and rest tremors. Although the molecular mechanism of PD is still not fully understood, neuroinflammation has a key role in the damage of dopaminergic neurons. Herein, we found that kurarinone, a unique natural product from Sophora flavescens, alleviated the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced behavioral deficits and dopaminergic neurotoxicity, including the losses of neurotransmitters and tyrosine hydroxylase (TH)-positive cells (SN and striatum [STR]). Furthermore, kurarinone attenuated the MPTP-mediated neuroinflammation via suppressing the activation of microglia involved in the nuclear factor kappa B signaling pathway. The proteomics result of the solvent-induced protein precipitation and thermal proteome profiling suggest that the soluble epoxide hydrolase (sEH) enzyme, which is associated with the neuroinflammation of PD, is a promising target of kurarinone. This is supported by the increase of plasma epoxyeicosatrienoic acids (sEH substrates) and the decrease of dihydroxyeicosatrienoic acids (sEH products), and the results of in vitro inhibition kinetics, surface plasmon resonance, and cocrystallization of kurarinone with sEH revealed that this natural compound is an uncompetitive inhibitor. In addition, sEH knockout (KO) attenuated the progression of PD, and sEH KO plus kurarinone did not further reduce the protection of PD in MPTP-induced PD mice. These findings suggest that kurarinone could be a potential natural candidate for the treatment of PD, possibly through sEH inhibition.
ESTHER : Sun_2022_Proc.Natl.Acad.Sci.U.S.A_119_
PubMedSearch : Sun_2022_Proc.Natl.Acad.Sci.U.S.A_119_
PubMedID: 35217618
Gene_locus related to this paper: human-EPHX2

Title : Aflatoxin B(1) exposure disrupts the intestinal immune function via a soluble epoxide hydrolase-mediated manner - Wang_2022_Ecotoxicol.Environ.Saf_249_114417
Author(s) : Wang W , Wang Y , Yang J , Wagner KM , Hwang SH , Cheng J , Singh N , Edwards P , Morisseau C , Zhang G , Panigrahy D , Hammock BD
Ref : Ecotoxicology & Environmental Safety , 249 :114417 , 2022
Abstract : Aflatoxin B(1) (AFB(1)) contamination in food and feed leads to severe global health problems. Acting as the frontier immunological barrier, the intestinal mucosa is constantly challenged by exposure to foodborne toxins such as AFB(1) via contaminated diets, but the detailed toxic mechanism and endogenous regulators of AFB(1) toxicity are still unclear. Here, we showed that AFB(1) disrupted intestinal immune function by suppressing macrophages, especially M2 macrophages, and antimicrobial peptide-secreting Paneth cells. Using an oxylipinomics approach, we identified that AFB(1) immunotoxicity is associated with decreased epoxy fatty acids, notably epoxyeicosatrienoic acids, and increased soluble epoxide hydrolase (sEH) levels in the intestine. Furthermore, sEH deficiency or inhibition rescued the AFB(1)-compromised intestinal immunity by restoring M2 macrophages as well as Paneth cells and their-derived lysozyme and alpha-defensin-3 in mice. Altogether, our study demonstrates that AFB(1) exposure impairs intestinal immunity, at least in part, in a sEH-mediated way. Moreover, the present study supports the potential application of pharmacological intervention by inhibiting the sEH enzyme in alleviating intestinal immunotoxicity and associated complications caused by AFB(1) global contamination.
ESTHER : Wang_2022_Ecotoxicol.Environ.Saf_249_114417
PubMedSearch : Wang_2022_Ecotoxicol.Environ.Saf_249_114417
PubMedID: 36525946

Title : Anti-Inflammatory Activity of Soluble Epoxide Hydrolase Inhibitors Based on Selenoureas Bearing an Adamantane Moiety - Burmistrov_2022_Int.J.Mol.Sci_23_
Author(s) : Burmistrov V , Morisseau C , Babkov DA , Golubeva T , Pitushkin D , Sokolova EV , Vasipov V , Kuznetsov Y , Bazhenov SV , Novoyatlova US , Bondarev NA , Manukhov IV , Osipova V , Berberova N , Spasov AA , Butov GM , Hammock BD
Ref : Int J Mol Sci , 23 : , 2022
Abstract : The inhibitory potency of the series of inhibitors of the soluble epoxide hydrolase (sEH) based on the selenourea moiety and containing adamantane and aromatic lipophilic groups ranges from 34.3 nM to 1.2 microM. The most active compound 5d possesses aliphatic spacers between the selenourea group and lipophilic fragments. Synthesized compounds were tested against the LPS-induced activation of primary murine macrophages. The most prominent anti-inflammatory activity, defined as a suppression of nitric oxide synthesis by LPS-stimulated macrophages, was demonstrated for compounds 4a and 5b. The cytotoxicity of the obtained substances was studied using human neuroblastoma and fibroblast cell cultures. Using these cell assays, the cytotoxic concentration for 4a was 4.7-18.4 times higher than the effective anti-inflammatory concentration. The genotoxicity and the ability to induce oxidative stress was studied using bacterial lux-biosensors. Substance 4a does not exhibit genotoxic properties, but it can cause oxidative stress at concentrations above 50 microM. Put together, the data showed the efficacy and safety of compound 4a.
ESTHER : Burmistrov_2022_Int.J.Mol.Sci_23_
PubMedSearch : Burmistrov_2022_Int.J.Mol.Sci_23_
PubMedID: 36142611

Title : Synthesis, In Vitro Profiling, and In Vivo Evaluation of Benzohomoadamantane-Based Ureas for Visceral Pain: A New Indication for Soluble Epoxide Hydrolase Inhibitors - Codony_2022_J.Med.Chem_65_13660
Author(s) : Codony S , Entrena JM , Calvo-Tusell C , Jora B , Gonzalez-Cano R , Osuna S , Corpas R , Morisseau C , Perez B , Barniol-Xicota M , Grinan-Ferre C , Perez C , Rodriguez-Franco MI , Martinez AL , Loza MI , Pallas M , Verhelst SHL , Sanfeliu C , Feixas F , Hammock BD , Brea J , Cobos EJ , Vazquez S
Ref : Journal of Medicinal Chemistry , 65 :13660 , 2022
Abstract : The soluble epoxide hydrolase (sEH) has been suggested as a pharmacological target for the treatment of several diseases, including pain-related disorders. Herein, we report further medicinal chemistry around new benzohomoadamantane-based sEH inhibitors (sEHI) in order to improve the drug metabolism and pharmacokinetics properties of a previous hit. After an extensive in vitro screening cascade, molecular modeling, and in vivo pharmacokinetics studies, two candidates were evaluated in vivo in a murine model of capsaicin-induced allodynia. The two compounds showed an anti-allodynic effect in a dose-dependent manner. Moreover, the most potent compound presented robust analgesic efficacy in the cyclophosphamide-induced murine model of cystitis, a well-established model of visceral pain. Overall, these results suggest painful bladder syndrome as a new possible indication for sEHI, opening a new range of applications for them in the visceral pain field.
ESTHER : Codony_2022_J.Med.Chem_65_13660
PubMedSearch : Codony_2022_J.Med.Chem_65_13660
PubMedID: 36222708

Title : Structure-activity relationship studies of benzothiazole-phenyl analogs as multi-target ligands to alleviate pain without affecting normal behavior - Angelia_2022_Prostaglandins.Other.Lipid.Mediat__106702
Author(s) : Angelia J , Weng X , Solomatov A , Chin C , Fernandez A , Hudson PK , Morisseau C , Hammock BD , Kandasamy R , Pecic S
Ref : Prostaglandins Other Lipid Mediat , :106702 , 2022
Abstract : Soluble epoxide hydrolase (sEH) and fatty acid amide hydrolase (FAAH) are potential targets for several diseases. Previous studies have reported that concomitant selective inhibition of sEH and FAAH produced antinociception effects in an animal model of pain. However, the co-administration of a selective sEH inhibitor and a selective FAAH inhibitor might produce serious side effects due to drug-drug interactions that could complicate drug development in the long term. Thus, discovering dual sEH/FAAH inhibitors, single small molecules that can simultaneously inhibit both sEH and FAAH, would be a significant accomplishment in the medicinal chemistry field. Herein, we report the synthesis and biological evaluation of benzothiazole-phenyl-based analogs as potential dual sEH/FAAH inhibitors. This work represents a follow-up structure-activity relationship (SAR) and metabolic-stability studies of our best dual sEH/FAAH inhibitor identified previously, as well as in vivo evaluation of its effects on voluntary locomotor behavior in rats. Our SAR study indicates that trifluoromethyl groups on the aromatic rings are well tolerated by the targeted enzymes when placed at the ortho and para positions; however, they, surprisingly, did not improve metabolic stability in liver microsomes. Our behavioral studies indicate that doses of dual sEH/FAAH inhibitors that alleviate pain do not depress voluntary behavior in naive rats, which is a common side effect of currently available analgesic drugs (e.g., opioids). Thus, dual sEH/FAAH inhibitors may be a safe and effective approach to treat pain.
ESTHER : Angelia_2022_Prostaglandins.Other.Lipid.Mediat__106702
PubMedSearch : Angelia_2022_Prostaglandins.Other.Lipid.Mediat__106702
PubMedID: 36529320

Title : Discovery and In Vivo Proof of Concept of a Highly Potent Dual Inhibitor of Soluble Epoxide Hydrolase and Acetylcholinesterase for the Treatment of Alzheimer's Disease - Codony_2022_J.Med.Chem_65_4909
Author(s) : Codony S , Pont C , Grinan-Ferre C , Di Pede-Mattatelli A , Calvo-Tusell C , Feixas F , Osuna S , Jarne-Ferrer J , Naldi M , Bartolini M , Loza MI , Brea J , Perez B , Bartra C , Sanfeliu C , Juarez-Jimenez J , Morisseau C , Hammock BD , Pallas M , Vazquez S , Munoz-Torrero D
Ref : Journal of Medicinal Chemistry , 65 :4909 , 2022
Abstract : With innumerable clinical failures of target-specific drug candidates for multifactorial diseases, such as Alzheimer's disease (AD), which remains inefficiently treated, the advent of multitarget drug discovery has brought a new breath of hope. Here, we disclose a class of 6-chlorotacrine (huprine)-TPPU hybrids as dual inhibitors of the enzymes soluble epoxide hydrolase (sEH) and acetylcholinesterase (AChE), a multitarget profile to provide cumulative effects against neuroinflammation and memory impairment. Computational studies confirmed the gorge-wide occupancy of both enzymes, from the main site to a secondary site, including a so far non-described AChE cryptic pocket. The lead compound displayed in vitro dual nanomolar potencies, adequate brain permeability, aqueous solubility, human microsomal stability, lack of neurotoxicity, and it rescued memory, synaptic plasticity, and neuroinflammation in an AD mouse model, after low dose chronic oral administration.
ESTHER : Codony_2022_J.Med.Chem_65_4909
PubMedSearch : Codony_2022_J.Med.Chem_65_4909
PubMedID: 35271276

Title : Discovery of memantyl urea derivatives as potent soluble epoxide hydrolase inhibitors against lipopolysaccharide-induced sepsis - Du_2021_Eur.J.Med.Chem_223_113678
Author(s) : Du F , Sun W , Morisseau C , Hammock BD , Bao X , Liu Q , Wang C , Zhang T , Yang H , Zhou J , Xiao W , Liu Z , Chen G
Ref : Eur Journal of Medicinal Chemistry , 223 :113678 , 2021
Abstract : Sepsis, a systemic inflammatory response, caused by pathogenic factors including microorganisms, has high mortality and limited therapeutic approaches. Herein, a new soluble epoxide hydrolase (sEH) inhibitor series comprising a phenyl ring connected to a memantyl moiety via a urea or amide linkage has been designed. A preferential urea pharmacophore that improved the binding properties of the compounds was identified for those series via biochemical assay in vitro and in vivo studies. Molecular docking displayed that 3,5-dimethyl on the adamantyl group in B401 could make van der Waals interactions with residues at a hydrophobic pocket of sEH active site, which might indirectly explain the subnanomolar level activities of memantyl urea derivatives in vitro better than AR-9281. Among them, compound B401 significantly improved the inhibition potency with human and murine sEH IC(50) values as 0.4 nM and 0.5 nM, respectively. Although the median survival time of C57BL/6 mice in LPS-induced sepsis model was slightly increased, the survival rate did not reach significant efficacy. Based on safety profile, metabolic stability, pharmacokinetic and in vivo efficacy, B401 demonstrated the proof of potential for this class of memantyl urea-based sEH inhibitors as therapeutic agents in sepsis.
ESTHER : Du_2021_Eur.J.Med.Chem_223_113678
PubMedSearch : Du_2021_Eur.J.Med.Chem_223_113678
PubMedID: 34218083

Title : Inhibition of the Soluble Epoxide Hydrolase as an Analgesic Strategy: A Review of Preclinical Evidence - Wang_2021_J.Pain.Res_14_61
Author(s) : Wang Y , Wagner KM , Morisseau C , Hammock BD
Ref : J Pain Res , 14 :61 , 2021
Abstract : Chronic pain is a complicated condition which causes substantial physical, emotional, and financial impacts on individuals and society. However, due to high cost, lack of efficacy and safety problems, current treatments are insufficient. There is a clear unmet medical need for safe, nonaddictive and effective therapies in the management of pain. Epoxy-fatty acids (EpFAs), which are natural signaling molecules, play key roles in mediation of both inflammatory and neuropathic pain sensation. However, their molecular mechanisms of action remain largely unknown. Soluble epoxide hydrolase (sEH) rapidly converts EpFAs into less bioactive fatty acid diols in vivo; therefore, inhibition of sEH is an emerging therapeutic target to enhance the beneficial effect of natural EpFAs. In this review, we will discuss sEH inhibition as an analgesic strategy for pain management and the underlying molecular mechanisms.
ESTHER : Wang_2021_J.Pain.Res_14_61
PubMedSearch : Wang_2021_J.Pain.Res_14_61
PubMedID: 33488116

Title : Ibuprofen alters epoxide hydrolase activity and epoxy-oxylipin metabolites associated with different metabolic pathways in murine livers - Tiwari_2021_Sci.Rep_11_7042
Author(s) : Tiwari S , Yang J , Morisseau C , Durbin-Johnson B , Hammock BD , Gomes AV
Ref : Sci Rep , 11 :7042 , 2021
Abstract : Over the last decade oxylipins have become more recognized for their involvement in several diseases. Non-steroidal anti-inflammatory drugs (NSAIDs) such as ibuprofen are known to inhibit cyclooxygenase (COX) enzymes, but how NSAIDs affect oxylipins, in addition to COX products, in animal tissues is not well understood. Oxylipins in livers from male and female mice treated with 100 mg/kg/day of ibuprofen for 7 days were investigated. The results showed that ibuprofen treated male livers contained 7 times more altered oxylipins than ibuprofen treated female livers. In male and female livers some prostaglandins were altered, while diols, hydroxy fatty acids and epoxides were significantly altered in male livers. Some soluble epoxide hydrolase (sEH) products, such as 9,10-DiHODE were found to be decreased, while sEH substrates (such as 9(10)-EpODE and 5(6)-EpETrE) were found to be increased in male livers treated with ibuprofen, but not in ibuprofen treated female livers. The enzymatic activities of sEH and microsomal epoxide hydrolase (mEH) were elevated by ibuprofen in both males and females. Analyzing the influence of sex on the effect of ibuprofen on oxylipins and COX products showed that approximately 27% of oxylipins detected were influenced by sex. The results reveal that ibuprofen disturbs not only the COX pathway, but also the CYP450 and lipoxygenase pathways in male mice, suggesting that ibuprofen is likely to generate sex related differences in biologically active oxylipins. Increased sEH activity after ibuprofen treatment is likely to be one of the mechanisms by which the liver reduces the higher levels of EpODEs and EpETrEs.
ESTHER : Tiwari_2021_Sci.Rep_11_7042
PubMedSearch : Tiwari_2021_Sci.Rep_11_7042
PubMedID: 33782432

Title : Soluble Epoxide Hydrolase Hepatic Deficiency Ameliorates Alcohol-Associated Liver Disease - Mello_2021_Cell.Mol.Gastroenterol.Hepatol_11_815
Author(s) : Mello A , Hsu MF , Koike S , Chu B , Cheng J , Yang J , Morisseau C , Torok NJ , Hammock BD , Haj FG
Ref : Cell Mol Gastroenterol Hepatol , 11 :815 , 2021
Abstract : BACKGROUND & AIMS: Alcohol-associated liver disease (ALD) is a significant cause of liver-related morbidity and mortality worldwide and with limited therapies. Soluble epoxide hydrolase (sEH; Ephx2) is a largely cytosolic enzyme that is highly expressed in the liver and is implicated in hepatic function, but its role in ALD is mostly unexplored. METHODS: To decipher the role of hepatic sEH in ALD, we generated mice with liver-specific sEH disruption (Alb-Cre; Ephx2(fl/fl)). Alb-Cre; Ephx2(fl/fl) and control (Ephx2(fl/fl)) mice were subjected to an ethanol challenge using the chronic plus binge model of ALD and hepatic injury, inflammation, and steatosis were evaluated under pair-fed and ethanol-fed states. In addition, we investigated the capacity of pharmacologic inhibition of sEH in the chronic plus binge mouse model. RESULTS: We observed an increase of hepatic sEH in mice upon ethanol consumption, suggesting that dysregulated hepatic sEH expression might be involved in ALD. Alb-Cre; Ephx2(fl/fl) mice presented efficient deletion of hepatic sEH with corresponding attenuation in sEH activity and alteration in the lipid epoxide/diol ratio. Consistently, hepatic sEH deficiency ameliorated ethanol-induced hepatic injury, inflammation, and steatosis. In addition, targeted metabolomics identified lipid mediators that were impacted significantly by hepatic sEH deficiency. Moreover, hepatic sEH deficiency was associated with a significant attenuation of ethanol-induced hepatic endoplasmic reticulum and oxidative stress. Notably, pharmacologic inhibition of sEH recapitulated the effects of hepatic sEH deficiency and abrogated injury, inflammation, and steatosis caused by ethanol feeding. CONCLUSIONS: These findings elucidated a role for sEH in ALD and validated a pharmacologic inhibitor of this enzyme in a preclinical mouse model as a potential therapeutic approach.
ESTHER : Mello_2021_Cell.Mol.Gastroenterol.Hepatol_11_815
PubMedSearch : Mello_2021_Cell.Mol.Gastroenterol.Hepatol_11_815
PubMedID: 33068774

Title : Inhibition of sEH via stabilizing the level of EETs alleviated Alzheimer's disease through GSK3beta signaling pathway - Sun_2021_Food.Chem.Toxicol_156_112516
Author(s) : Sun CP , Zhang XY , Zhou JJ , Huo XK , Yu ZL , Morisseau C , Hammock BD , Ma XC
Ref : Food & Chemical Toxicology , 156 :112516 , 2021
Abstract : Alzheimer's disease (AD) is the most common neurodegenerative disorder characterized by dementia. Inhibition of soluble epoxide hydrolase (sEH) regulates inflammation involving in central nervous system (CNS) diseases. However, the exactly mechanism of sEH in AD is still unclear. In this study, we evaluated the vital role of sEH in amyloid beta (Abeta)-induced AD mice, and revealed a possible molecular mechanism for inhibition of sEH in the treatment of AD. The results showed that the sEH expression and activity were remarkably increased in the hippocampus of Abeta-induced AD mice. Chemical inhibition of sEH by TPPU, a selective sEH inhibitor, alleviated spatial learning and memory deficits, and elevated levels of neurotransmitters in Abeta-induced AD mice. Furthermore, inhibition of sEH could ameliorate neuroinflammation, neuronal death, and oxidative stress via stabilizing the in vivo level of epoxyeicosatrienoic acids (EETs), especially 8,9-EET and 14,15-EET, further resulting in the anti-AD effect through the regulation of GSK3beta-mediated NF-kappaB, p53, and Nrf2 signaling pathways. These findings revealed the underlying mechanism of sEH as a potential therapeutic target in treatment of AD.
ESTHER : Sun_2021_Food.Chem.Toxicol_156_112516
PubMedSearch : Sun_2021_Food.Chem.Toxicol_156_112516
PubMedID: 34411643

Title : Soluble Epoxide Hydrolase Deletion Limits High-Fat Diet-Induced Inflammation - Wagner_2021_Front.Pharmacol_12_778470
Author(s) : Wagner KM , Yang J , Morisseau C , Hammock BD
Ref : Front Pharmacol , 12 :778470 , 2021
Abstract : The soluble epoxide hydrolase (sEH) enzyme is a major regulator of bioactive lipids. The enzyme is highly expressed in liver and kidney and modulates levels of endogenous epoxy-fatty acids, which have pleiotropic biological effects including limiting inflammation, neuroinflammation, and hypertension. It has been hypothesized that inhibiting sEH has beneficial effects on limiting obesity and metabolic disease as well. There is a body of literature published on these effects, but typically only male subjects have been included. Here, we investigate the role of sEH in both male and female mice and use a global sEH knockout mouse model to compare the effects of diet and diet-induced obesity. The results demonstrate that sEH activity in the liver is modulated by high-fat diets more in male than in female mice. In addition, we characterized the sEH activity in high fat content tissues and demonstrated the influence of diet on levels of bioactive epoxy-fatty acids. The sEH KO animals had generally increased epoxy-fatty acids compared to wild-type mice but gained less body weight on higher-fat diets. Generally, proinflammatory prostaglandins and triglycerides were also lower in livers of sEH KO mice fed HFD. Thus, sEH activity, prostaglandins, and triglycerides increase in male mice on high-fat diet but are all limited by sEH ablation. Additionally, these changes also occur in female mice though at a different magnitude and are also improved by knockout of the sEH enzyme.
ESTHER : Wagner_2021_Front.Pharmacol_12_778470
PubMedSearch : Wagner_2021_Front.Pharmacol_12_778470
PubMedID: 34975481
Gene_locus related to this paper: mouse-hyes

Title : Species Differences in Metabolism of Soluble Epoxide Hydrolase Inhibitor, EC1728, Highlight the Importance of Clinically Relevant Screening Mechanisms in Drug Development - McReynolds_2021_Molecules_26_
Author(s) : McReynolds CB , Yang J , Guedes A , Morisseau C , Garcia R , Knych H , Tearney C , Hamamoto B , Hwang SH , Wagner K , Hammock BD
Ref : Molecules , 26 : , 2021
Abstract : There are few novel therapeutic options available for companion animals, and medications rely heavily on repurposed drugs developed for other species. Considering the diversity of species and breeds in companion animal medicine, comprehensive PK exposures in the companion animal patient is often lacking. The purpose of this paper was to assess the pharmacokinetics after oral and intravenous dosing in domesticated animal species (dogs, cats, and horses) of a novel soluble epoxide hydrolase inhibitor, EC1728, being developed for the treatment of pain in animals. Results: Intravenous and oral administration revealed that bioavailability was similar for dogs, and horses (42 and 50% F) but lower in mice and cats (34 and 8%, respectively). Additionally, clearance was similar between cats and mice, but >2x faster in cats vs. dogs and horses. Efficacy with EC1728 has been demonstrated in mice, dogs, and horses, and despite the rapid clearance of EC1728 in cats, analgesic efficacy was demonstrated in an acute pain model after intravenous but not oral dosing. Conclusion: These results demonstrate that exposures across species can vary, and investigation of therapeutic exposures in target species is needed to provide adequate care that addresses efficacy and avoids toxicity.
ESTHER : McReynolds_2021_Molecules_26_
PubMedSearch : McReynolds_2021_Molecules_26_
PubMedID: 34443621

Title : Activity of sEH and Oxidant Status during Systemic Bovine Coliform Mastitis - Mavangira_2021_Antioxidants.(Basel)_10_
Author(s) : Mavangira V , Kuhn MJ , Abuelo A , Morisseau C , Hammock BD , Sordillo LM
Ref : Antioxidants (Basel) , 10 : , 2021
Abstract : Bovine coliform mastitis presents treatment challenges because of systemic inflammation and oxidative stress. Soluble epoxide hydrolase (sEH) is a promising therapeutic target in conditions characterized by inflammation and oxidative stress but has not been evaluated in cattle. We compared sEH activity and oxidant status in healthy Holstein dairy cows to those with systemic coliform mastitis (n = 5/group) using complementary approaches. First, the activity of sEH on [(3)H]-trans-diphenyl-propene oxide (tDPPO) was assessed ex vivo using tissue homogenates (mammary, liver, and kidney). Second, the concentrations of sEH substrates and metabolites in plasma, milk, and urine were determined as an index of in vivo sEH activity. Oxidant status was assessed in serum and milk. Data were analyzed by non-parametric methods. Metabolism of tDPPO was greater in mammary tissues from cows with coliform mastitis compared to controls. In contrast, ratios of sEH substrates and metabolites predicted lower sEH activity in cows with coliform mastitis than controls. Milk oxidant status showed greater prooxidant levels in coliform mastitis cows. Cows with coliform mastitis exhibit increased sEH activity in mammary tissue; at the same time, milk oxidant status is increased. Future studies should characterize sEH activity and oxidant status patterns and explore therapies targeting sEH during coliform mastitis.
ESTHER : Mavangira_2021_Antioxidants.(Basel)_10_
PubMedSearch : Mavangira_2021_Antioxidants.(Basel)_10_
PubMedID: 34065244

Title : Relative Importance of Soluble and Microsomal Epoxide Hydrolases for the Hydrolysis of Epoxy-Fatty Acids in Human Tissues - Morisseau_2021_Int.J.Mol.Sci_22_
Author(s) : Morisseau C , Kodani SD , Kamita SG , Yang J , Lee KSS , Hammock BD
Ref : Int J Mol Sci , 22 : , 2021
Abstract : Epoxy-fatty acids (EpFAs) are endogenous lipid mediators that have a large breadth of biological activities, including the regulation of blood pressure, inflammation, angiogenesis, and pain perception. For the past 20 years, soluble epoxide hydrolase (sEH) has been recognized as the primary enzyme for degrading EpFAs in vivo. The sEH converts EpFAs to the generally less biologically active 1,2-diols, which are quickly eliminated from the body. Thus, inhibitors of sEH are being developed as potential drug therapeutics for various diseases including neuropathic pain. Recent findings suggest that other epoxide hydrolases (EHs) such as microsomal epoxide hydrolase (mEH) and epoxide hydrolase-3 (EH3) can contribute significantly to the in vivo metabolism of EpFAs. In this study, we used two complementary approaches to probe the relative importance of sEH, mEH, and EH3 in 15 human tissue extracts: hydrolysis of 14,15-EET and 13,14-EDP using selective inhibitors and protein quantification. The sEH hydrolyzed the majority of EpFAs in all of the tissues investigated, mEH hydrolyzed a significant portion of EpFAs in several tissues, whereas no significant role in EpFAs metabolism was observed for EH3. Our findings indicate that residual mEH activity could limit the therapeutic efficacy of sEH inhibition in certain organs.
ESTHER : Morisseau_2021_Int.J.Mol.Sci_22_
PubMedSearch : Morisseau_2021_Int.J.Mol.Sci_22_
PubMedID: 34066758

Title : Adrenic Acid-Derived Epoxy Fatty Acids Are Naturally Occurring Lipids and Their Methyl Ester Prodrug Reduces Endoplasmic Reticulum Stress and Inflammatory Pain - Singh_2021_ACS.Omega_6_7165
Author(s) : Singh N , Barnych B , Wagner KM , Wan D , Morisseau C , Hammock BD
Ref : ACS Omega , 6 :7165 , 2021
Abstract : Adrenic acid (AdA, 22:4) is an omega-6 polyunsaturated fatty acid (PUFA), derived from arachidonic acid. Like other PUFAs, it is metabolized by cytochrome P450s to a group of epoxy fatty acids (EpFAs), epoxydocosatrienoic acids (EDTs). EpFAs are lipid mediators with various beneficial bioactivities, including exertion of analgesia and reduction of endoplasmic reticulum (ER) stress, that are degraded to dihydroxy fatty acids by the soluble epoxide hydrolase (sEH). However, the biological characteristics and activities of EDTs are relatively unexplored, and, alongside dihydroxydocosatrienoic acids (DHDTs), they had not been detected in vivo. Herein, EDT and DHDT regioisomers were synthesized, purified, and used as standards for analysis with a selective and quantitative high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method. Biological verification in AdA-rich tissues suggests that basal metabolite levels are highest in the liver, with 16,17-EDT concentrations consistently being the greatest across the analyzed tissues. Enzyme hydrolysis assessment revealed that EDTs are sEH substrates, with greatest relative rate preference for the 13,14-EDT regioisomer. Pretreatment with an EDT methyl ester regioisomer mixture significantly reduced the onset of tunicamycin-stimulated ER stress in human embryonic kidney cells. Finally, administration of the regioisomeric mixture effectively alleviated carrageenan-induced inflammatory pain in rats. This study indicates that EDTs and DHDTs are naturally occurring lipids, and EDTs could be another therapeutically relevant group of EpFAs.
ESTHER : Singh_2021_ACS.Omega_6_7165
PubMedSearch : Singh_2021_ACS.Omega_6_7165
PubMedID: 33748630

Title : From the Design to the In Vivo Evaluation of Benzohomoadamantane-Derived Soluble Epoxide Hydrolase Inhibitors for the Treatment of Acute Pancreatitis - Codony_2021_J.Med.Chem__
Author(s) : Codony S , Calvo-Tusell C , Valverde E , Osuna S , Morisseau C , Loza MI , Brea J , Perez C , Rodriguez-Franco MI , Pizarro-Delgado J , Corpas R , Grinan-Ferre C , Pallas M , Sanfeliu C , Vazquez-Carrera M , Hammock BD , Feixas F , Vazquez S
Ref : Journal of Medicinal Chemistry , : , 2021
Abstract : The pharmacological inhibition of soluble epoxide hydrolase (sEH) is efficient for the treatment of inflammatory and pain-related diseases. Numerous potent sEH inhibitors (sEHIs) present adamantyl or phenyl moieties, such as the clinical candidates AR9281 or EC5026. Herein, in a new series of sEHIs, these hydrophobic moieties have been merged in a benzohomoadamantane scaffold. Most of the new sEHIs have excellent inhibitory activities against sEH. Molecular dynamics simulations suggested that the addition of an aromatic ring into the adamantane scaffold produced conformational rearrangements in the enzyme to stabilize the aromatic ring of the benzohomoadamantane core. A screening cascade permitted us to select a candidate for an in vivo efficacy study in a murine model of cerulein-induced acute pancreatitis. The administration of 22 improved the health status of the animals and reduced pancreatic damage, demonstrating that the benzohomoadamantane unit is a promising scaffold for the design of novel sEHIs.
ESTHER : Codony_2021_J.Med.Chem__
PubMedSearch : Codony_2021_J.Med.Chem__
PubMedID: 33945278

Title : Genetic deficiency or pharmacological inhibition of soluble epoxide hydrolase ameliorates high fat diet-induced pancreatic beta-cell dysfunction and loss - Koike_2021_Free.Radic.Biol.Med__
Author(s) : Koike S , Hsu MF , Bettaieb A , Chu B , Matsumoto N , Morisseau C , Havel PJ , Huising MO , Hammock BD , Haj FG
Ref : Free Radic Biol Med , : , 2021
Abstract : Pancreatic beta-cells are crucial regulators of systemic glucose homeostasis, and their dysfunction and loss are a central feature in type 2 diabetes. Interventions that rectify beta-cell dysfunction and loss are essential to combat this deadly malady. In the current study, we sought to delineate the role of soluble epoxide hydrolase (sEH) in beta-cells under diet-induced metabolic stress. The expression of sEH was upregulated in murine and macaque diabetes models and islets of diabetic human patients. We postulated that hyperglycemia-induced elevation in sEH leads to a reduction in its substrates, epoxyeicosatrienoic acids (EETs), and attenuates the function of beta-cells. Genetic deficiency of sEH potentiated glucose-stimulated insulin secretion in mice, likely in a cell-autonomous manner, contributing to better systemic glucose control. Consistent with this observation, genetic and pharmacological inactivation of sEH and the treatment with EETs exhibited insulinotropic effects on isolated murine islets ex vivo. Additionally, sEH deficiency enhanced glucose sensing and metabolism with elevated ATP and cAMP concentrations. This phenotype was associated with attenuated oxidative stress and diminished beta-cell death in sEH deficient islets. Moreover, pharmacological inhibition of sEH in vivo mitigated, albeit partly, high fat diet-induced beta-cell loss and dedifferentiation. The current observations provide new insights into the role of sEH in beta-cells and information that may be leveraged for the development of a mechanism-based intervention to rectify beta-cell dysfunction and loss.
ESTHER : Koike_2021_Free.Radic.Biol.Med__
PubMedSearch : Koike_2021_Free.Radic.Biol.Med__
PubMedID: 34038767

Title : EPHX1 mutations cause a lipoatrophic diabetes syndrome due to impaired epoxide hydrolysis and increased cellular senescence - Gautheron_2021_Elife_10_
Author(s) : Gautheron J , Morisseau C , Chung WK , Zammouri J , Auclair M , Baujat G , Capel E , Moulin C , Wang Y , Yang J , Hammock BD , Cerame B , Phan F , Feve B , Vigouroux C , Andreelli F , Jeru I
Ref : Elife , 10 : , 2021
Abstract : Epoxide hydrolases (EHs) regulate cellular homeostasis through hydrolysis of epoxides to less-reactive diols. The first discovered EH was EPHX1, also known as mEH. EH functions remain partly unknown, and no pathogenic variants have been reported in humans. We identified two de novo variants located in EPHX1 catalytic site in patients with a lipoatrophic diabetes characterized by loss of adipose tissue, insulin resistance, and multiple organ dysfunction. Functional analyses revealed that these variants led to the protein aggregation within the endoplasmic reticulum and to a loss of its hydrolysis activity. CRISPR-Cas9-mediated EPHX1 knockout (KO) abolished adipocyte differentiation and decreased insulin response. This KO also promoted oxidative stress and cellular senescence, an observation confirmed in patient-derived fibroblasts. Metreleptin therapy had a beneficial effect in one patient. This translational study highlights the importance of epoxide regulation for adipocyte function and provides new insights into the physiological roles of EHs in humans.
ESTHER : Gautheron_2021_Elife_10_
PubMedSearch : Gautheron_2021_Elife_10_
PubMedID: 34342583
Gene_locus related to this paper: human-EPHX1

Title : New Alkoxy- Analogues of Epoxyeicosatrienoic Acids Attenuate Cisplatin Nephrotoxicity In Vitro via Reduction of Mitochondrial Dysfunction, Oxidative Stress, Mitogen-Activated Protein Kinase Signaling, and Caspase Activation - Singh_2021_Chem.Res.Toxicol__
Author(s) : Singh N , Vik A , Lybrand DB , Morisseau C , Hammock BD
Ref : Chemical Research in Toxicology , : , 2021
Abstract : The usage of cisplatin, a highly potent chemotherapeutic, is limited by its severe nephrotoxicity. Arachidonic acid (ARA)-derived epoxyeicosatrienoic acids (EETs) and soluble epoxide hydrolase (sEH) inhibitors were shown to ameliorate this dose-limiting side effect, but both approaches have some pharmacological limitations. Analogues of EETs are an alternative avenue with unique benefits, but the current series of analogues face concerns regarding their structure and mimetic functionality. Hence, in this study, regioisomeric mixtures of four new ARA alkyl ethers were synthesized, characterized, and assessed as EET analogues against the concentration- and time-dependent toxicities of cisplatin in porcine proximal tubular epithelial cells. All four ether groups displayed bioisostere activity, ranging from marginal for methoxy- (1), good for n-propoxy- (4), and excellent for ethoxy- (2) and i-propoxy- (3). Compounds 2 and 3 displayed cytoprotective effects comparable to that of an EET regioisomeric mixture (5) against high, acute cisplatin exposures but were more potent against low to moderate, chronic exposures. Compounds 2 and 3 (and 5) acted through stabilization of the mitochondrial transmembrane potential and attenuation of reactive oxygen species, leading to reduced phosphorylation of mitogen-activated protein kinases p38 and JNK and decreased activation of caspase-9 and caspase-3. This study demonstrates that alkoxy- groups are potent and more metabolically stable bioisostere alternatives to the epoxide within EETs that enable sEH-independent activity. It also illustrates the potential of ether-based mimics of EETs and other epoxy fatty acids as promising nephroprotective agents to tackle the clinically relevant side effect of cisplatin without compromising its antineoplastic function.
ESTHER : Singh_2021_Chem.Res.Toxicol__
PubMedSearch : Singh_2021_Chem.Res.Toxicol__
PubMedID: 34817988

Title : Discovery of Soluble Epoxide Hydrolase Inhibitors from Chemical Synthesis and Natural Products - Sun_2021_J.Med.Chem_64_184
Author(s) : Sun CP , Zhang XY , Morisseau C , Hwang SH , Zhang ZJ , Hammock BD , Ma XC
Ref : Journal of Medicinal Chemistry , 64 :184 , 2021
Abstract : Soluble epoxide hydrolase (sEH) is an alpha/beta hydrolase fold protein and widely distributed in numerous organs including the liver, kidney, and brain. The inhibition of sEH can effectively maintain endogenous epoxyeicosatrienoic acids (EETs) levels and reduce dihydroxyeicosatrienoic acids (DHETs) levels, resulting in therapeutic potentials for cardiovascular, central nervous system, and metabolic diseases. Therefore, since the beginning of this century, the development of sEH inhibitors is a hot research topic. A variety of potent sEH inhibitors have been developed by chemical synthesis or isolated from natural sources. In this review, we mainly summarized the interconnected aspects of sEH with cardiovascular, central nervous system, and metabolic diseases and then focus on representative inhibitors, which would provide some useful guidance for the future development of potential sEH inhibitors.
ESTHER : Sun_2021_J.Med.Chem_64_184
PubMedSearch : Sun_2021_J.Med.Chem_64_184
PubMedID: 33369424

Title : Differential Effects of 17,18-EEQ and 19,20-EDP Combined with Soluble Epoxide Hydrolase Inhibitor t-TUCB on Diet-Induced Obesity in Mice - Yang_2021_Int.J.Mol.Sci_22_
Author(s) : Yang Y , Xu X , Wu H , Yang J , Chen J , Morisseau C , Hammock BD , Bettaieb A , Zhao L
Ref : Int J Mol Sci , 22 : , 2021
Abstract : 17,18-Epoxyeicosatetraenoic acid (17,18-EEQ) and 19,20-epoxydocosapentaenoic acid (19,20-EDP) are bioactive epoxides produced from n-3 polyunsaturated fatty acid eicosapentaenoic acid and docosahexaenoic acid, respectively. However, these epoxides are quickly metabolized into less active diols by soluble epoxide hydrolase (sEH). We have previously demonstrated that an sEH inhibitor, t-TUCB, decreased serum triglycerides (TG) and increased lipid metabolic protein expression in the brown adipose tissue (BAT) of diet-induced obese mice. This study investigates the preventive effects of t-TUCB (T) alone or combined with 19,20-EDP (T + EDP) or 17,18-EEQ (T + EEQ) on BAT activation in the development of diet-induced obesity and metabolic disorders via osmotic minipump delivery in mice. Both T + EDP and T + EEQ groups showed significant improvement in fasting glucose, serum triglycerides, and higher core body temperature, whereas heat production was only significantly increased in the T + EEQ group. Moreover, both the T + EDP and T + EEQ groups showed less lipid accumulation in the BAT. Although UCP1 expression was not changed, PGC1alpha expression was increased in all three treated groups. In contrast, the expression of CPT1A and CPT1B, which are responsible for the rate-limiting step for fatty acid oxidation, was only increased in the T + EDP and T + EEQ groups. Interestingly, as a fatty acid transporter, CD36 expression was only increased in the T + EEQ group. Furthermore, both the T + EDP and T + EEQ groups showed decreased inflammatory NFkappaB signaling in the BAT. Our results suggest that 17,18-EEQ or 19,20-EDP combined with t-TUCB may prevent high-fat diet-induced metabolic disorders, in part through increased thermogenesis, upregulating lipid metabolic protein expression, and decreasing inflammation in the BAT.
ESTHER : Yang_2021_Int.J.Mol.Sci_22_
PubMedSearch : Yang_2021_Int.J.Mol.Sci_22_
PubMedID: 34361032

Title : Further exploration of the structure-activity relationship of dual soluble epoxide hydrolase\/fatty acid amide hydrolase inhibitors - Wilt_2021_Bioorg.Med.Chem_51_116507
Author(s) : Wilt S , Kodani S , Valencia L , Hudson PK , Sanchez S , Quintana T , Morisseau C , Hammock BD , Kandasamy R , Pecic S
Ref : Bioorganic & Medicinal Chemistry , 51 :116507 , 2021
Abstract : Fatty acid amide hydrolase (FAAH) is a membrane protein that hydrolyzes endocannabinoids, and its inhibition produces analgesic and anti-inflammatory effects. The soluble epoxide hydrolase (sEH) hydrolyzes epoxyeicosatrienoic acids (EETs) to dihydroxyeicosatetraenoic acids. EETs have anti-inflammatory and inflammation resolving properties, thus inhibition of sEH consequently reduces inflammation. Concurrent inhibition of both enzymes may represent a novel approach in the treatment of chronic pain. Drugs with multiple targets can provide a superior therapeutic effect and a decrease in side effects compared to ligands with single targets. Previously, microwave-assisted methodologies were employed to synthesize libraries of benzothiazole analogs from which high affinity dual inhibitors (e.g. 3, sEH IC(50) = 9.6 nM; FAAH IC(50) = 7 nM) were identified. Here, our structure-activity relationship studies revealed that the 4-phenylthiazole moiety is well tolerated by both enzymes, producing excellent inhibition potencies in the low nanomolar range (e.g. 6o, sEH IC(50) = 2.5 nM; FAAH IC(50) = 9.8 nM). Docking experiments show that the new class of dual inhibitors bind within the catalytic sites of both enzymes. Prediction of several pharmacokinetic/pharmacodynamic properties suggest that these new dual inhibitors are good candidates for further in vivo evaluation. Finally, dual inhibitor 3 was tested in the Formalin Test, a rat model of acute inflammatory pain. The data indicate that 3 produces antinociception against the inflammatory phase of the Formalin Test in vivo and is metabolically stable following intraperitoneal administration in male rats. Further, antinociception produced by 3 is comparable to that of ketoprofen, a traditional nonsteroidal anti-inflammatory drug. The results presented here will help toward the long-term goal of developing novel non-opioid therapeutics for pain management.
ESTHER : Wilt_2021_Bioorg.Med.Chem_51_116507
PubMedSearch : Wilt_2021_Bioorg.Med.Chem_51_116507
PubMedID: 34794001

Title : Role of epoxide hydrolases and cytochrome P450s on metabolism of KZR-616, a first-in-class selective inhibitor of the immunoproteasome - Fang_2021_Drug.Metab.Dispos__
Author(s) : Fang Y , Johnson H , Anderl JL , Muchamuel T , McMinn D , Morisseau C , Hammock BD , Kirk C , Wang J
Ref : Drug Metabolism & Disposition: The Biological Fate of Chemicals , : , 2021
Abstract : KZR-616 is an irreversible tripeptide epoxyketone-based selective inhibitor of the human immunoproteasome. Inhibition of the immunoproteasome results in anti-inflammatory activity in vitro and, based on promising therapeutic activity in animal models of rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE), KZR-616 is being developed for potential treatment of multiple autoimmune and inflammatory diseases. The presence of a ketoepoxide pharmacophore presents unique challenges in the study of drug metabolism during lead optimization and clinical candidate profiling. This study presents a thorough and systematic in vitro and cell-based enzymatic metabolism and kinetic investigation to identify the major enzymes involved in the metabolism and elimination of KZR-616. Upon exposure to liver microsomes in the absence of NADPH, KZR-616 and its analogs were converted to their inactive diol derivatives with varying degrees of stability. Diol formation was also shown to be the major metabolite in pharmacokinetic studies in monkeys and correlated with in vitro stability results for individual compounds. Further study in intact hepatocytes and a hepatocellular carcinoma cell line revealed that KZR-616 metabolism was sensitive to an inhibitor of microsomal epoxide hydrolase (mEH) but not inhibitors of cytochrome P450 (CYP) or soluble epoxide hydrolase (sEH). Primary human hepatocytes were determined to be the most robust source of mEH activity for study in vitro These findings also suggest that the exposure of KZR-616 in vivo is unlikely to be affected by co-administration of inhibitors or inducers of CYP and sEH. Significance Statement This work presents a thorough and systematic investigation of metabolism and kinetic of KZR-616 and other peptide epoxyketones in in vitro and cell-based enzymatic systems. Gained information could be useful in assessing novel covalent proteasome inhibitors during lead compound optimization. The study also demonstrates a robust source of in vitro metabolism identification that correlated very well with in vivo PK metabolism for peptide epoxyketones.
ESTHER : Fang_2021_Drug.Metab.Dispos__
PubMedSearch : Fang_2021_Drug.Metab.Dispos__
PubMedID: 34234005
Gene_locus related to this paper: human-EPHX1

Title : 2-(Piperidin-4-yl)acetamides as Potent Inhibitors of Soluble Epoxide Hydrolase with Anti-Inflammatory Activity - Martin-Lopez_2021_Pharmaceuticals.(Basel)_14_
Author(s) : Martin-Lopez J , Codony S , Bartra C , Morisseau C , Loza MI , Sanfeliu C , Hammock BD , Brea J , Vazquez S
Ref : Pharmaceuticals (Basel) , 14 : , 2021
Abstract : The pharmacological inhibition of soluble epoxide hydrolase (sEH) has been suggested as a potential therapy for the treatment of pain and inflammatory diseases through the stabilization of endogenous epoxyeicosatrienoic acids. Numerous potent sEH inhibitors (sEHI) have been developed, however many contain highly lipophilic substituents limiting their availability. Recently, a new series of benzohomoadamantane-based ureas endowed with potent inhibitory activity for the human and murine sEH was reported. However, their very low microsomal stability prevented further development. Herein, a new series of benzohomoadamantane-based amides were synthetized, fully characterized, and evaluated as sEHI. Most of these amides were endowed with excellent inhibitory potencies. A selected compound displayed anti-inflammatory effects with higher effectiveness than the reference sEHI, TPPU.
ESTHER : Martin-Lopez_2021_Pharmaceuticals.(Basel)_14_
PubMedSearch : Martin-Lopez_2021_Pharmaceuticals.(Basel)_14_
PubMedID: 34959721

Title : Pharmacological Inhibition of Soluble Epoxide Hydrolase as a New Therapy for Alzheimer's Disease - Grinan-Ferre_2020_Neurotherapeutics__
Author(s) : Grinan-Ferre C , Codony S , Pujol E , Yang J , Leiva R , Escolano C , Puigoriol-Illamola D , Companys-Alemany J , Corpas R , Sanfeliu C , Perez B , Loza MI , Brea J , Morisseau C , Hammock BD , Vazquez S , Pallas M , Galdeano C
Ref : Neurotherapeutics , : , 2020
Abstract : The inhibition of the enzyme soluble epoxide hydrolase (sEH) has demonstrated clinical therapeutic effects in several peripheral inflammatory-related diseases, with 3 compounds in clinical trials. However, the role of this enzyme in the neuroinflammation process has been largely neglected. Herein, we disclose the pharmacological validation of sEH as a novel target for the treatment of Alzheimer's disease (AD). Evaluation of cognitive impairment and pathological hallmarks were used in 2 models of age-related cognitive decline and AD using 3 structurally different and potent sEH inhibitors as chemical probes. sEH is upregulated in brains from AD patients. Our findings supported the beneficial effects of central sEH inhibition, regarding reducing cognitive impairment, neuroinflammation, tau hyperphosphorylation pathology, and the number of amyloid plaques. This study suggests that inhibition of inflammation in the brain by targeting sEH is a relevant therapeutic strategy for AD.
ESTHER : Grinan-Ferre_2020_Neurotherapeutics__
PubMedSearch : Grinan-Ferre_2020_Neurotherapeutics__
PubMedID: 32488482

Title : Soluble epoxide hydrolase inhibitor, TPPU, increases regulatory T cells pathway in an arthritis model - Trindade-da-Silva_2020_FASEB.J__
Author(s) : Trindade-da-Silva CA , Clemente-Napimoga JT , Abdalla HB , Rosa SM , Ueira-Vieira C , Morisseau C , Verri WA, Jr. , Montalli VAM , Hammock BD , Napimoga MH
Ref : FASEB Journal , : , 2020
Abstract : Epoxyeicosatrienoic acids (EET) and related epoxy fatty acids (EpFA) are endogenous anti-inflammatory compounds, which are converted by the soluble epoxide hydrolase (sEH) to dihydroxylethersatrienoic acids (DHETs) with lessened biological effects. Inhibition of sEH is used as a strategy to increase EET levels leading to lower inflammation. Rheumatoid arthritis is a chronic autoimmune disease that leads to destruction of joint tissues. This pathogenesis involves a complex interplay between the immune system, and environmental factors. Here, we investigate the effects of inhibiting sEH with 1-trifluoromethoxyphenyl-3-(1-propionylpiperidin-4-yl) urea (TPPU) on a collagen-induced arthritis model. The treatment with TPPU ameliorates hyperalgesia, edema, and decreases the expression of important pro-inflammatory cytokines of Th1 and Th17 profiles, while increasing Treg cells. Considering the challenges to control RA, this study provides robust data supporting that inhibition of the sEH is a promising target to treat arthritis.
ESTHER : Trindade-da-Silva_2020_FASEB.J__
PubMedSearch : Trindade-da-Silva_2020_FASEB.J__
PubMedID: 32400048

Title : Protostane-type triterpenoids as natural soluble epoxide hydrolase inhibitors: Inhibition potentials and molecular dynamics - Sun_2020_Bioorg.Chem_96_103637
Author(s) : Sun CP , Zhang J , Zhao WY , Yi J , Yan JK , Wang YL , Morisseau C , Liu ZB , Hammock BD , Ma XC
Ref : Bioorg Chem , 96 :103637 , 2020
Abstract : The inhibition of soluble epoxide hydrolase (sEH) is a promising therapeutic approach to treat inflammation and other disorders. In our present investigation on searching for sEH inhibitors from traditional Chinese medicines, we found that Alisma orientale displayed inhibition of sEH. We constructed a small library of protostane-type triterpenoids (1-25) isolated from A. orientale, and screened their inhibitory activities. Alismanin B (1), 11-deoxy-25-anhydro alisol E (4), 11-deoxy alisol B (5), and 25-O-ethyl alisol A (15) displayed concentration-dependently inhibitory activities against sEH with IC50 values from 3.40 +/- 0.57 muM to 9.57 +/- 0.88 muM. 11-Deoxy-25-anhydro alisol E (4) and 11-deoxy alisol B (5) were defined as mixed-type competitive inhibitors with Ki values of 12.6 and 3.48 muM, respectively, based on the result of inhibition kinetics. The potential interaction mechanism of 11-deoxy alisol B (5) with sEH was analyzed by molecular docking and molecular dynamics, revealing that amino acid residues Trp336 and Tyr466 were vital for its inhibitory activity.
ESTHER : Sun_2020_Bioorg.Chem_96_103637
PubMedSearch : Sun_2020_Bioorg.Chem_96_103637
PubMedID: 32032849

Title : Soluble Epoxide Hydrolase Inhibition by t-TUCB Promotes Brown Adipogenesis and Reduces Serum Triglycerides in Diet-Induced Obesity - Overby_2020_Int.J.Mol.Sci_21_
Author(s) : Overby H , Yang Y , Xu X , Graham K , Hildreth K , Choi S , Wan D , Morisseau C , Zeldin DC , Hammock BD , Wang S , Bettaieb A , Zhao L
Ref : Int J Mol Sci , 21 : , 2020
Abstract : Brown adipose tissue (BAT) is an important target for obesity treatment and prevention. Soluble epoxide hydrolase (sEH) converts bioactive epoxy fatty acids (EpFAs) into less active diols. sEH inhibitors (sEHI) are beneficial in many chronic diseases by stabilizing EpFAs. However, roles of sEH and sEHI in brown adipogenesis and BAT activity in treating diet-induced obesity (DIO) have not been reported. sEH expression was studied in in vitro models of brown adipogenesis and the fat tissues of DIO mice. The effects of the sEHI, trans-4-{4-[3-(4-trifluoromethoxy-phenyl)-ureido]-cyclohexyloxy-benzoic acid (t-TUCB), were studied in vitro and in the obese mice via mini osmotic pump delivery. sEH expression was increased in brown adipogenesis and the BAT of the DIO mice. t-TUCB promoted brown adipogenesis in vitro. Although t-TCUB did not change body weight, fat pad weight, or glucose and insulin tolerance in the obese mice, it decreased serum triglycerides and increased protein expression of genes important for lipid metabolism in the BAT. Our results suggest that sEH may play a critical role in brown adipogenesis, and sEHI may be beneficial in improving BAT protein expression involved in lipid metabolism. Further studies using the sEHI combined with EpFA generating diets for obesity treatment and prevention are warranted.
ESTHER : Overby_2020_Int.J.Mol.Sci_21_
PubMedSearch : Overby_2020_Int.J.Mol.Sci_21_
PubMedID: 32987880

Title : Selection of Potent Inhibitors of Soluble Epoxide Hydrolase for Usage in Veterinary Medicine - Shihadih_2020_Front.Vet.Sci_7_580
Author(s) : Shihadih DS , Harris TR , Kodani SD , Hwang SH , Lee KSS , Mavangira V , Hamamoto B , Guedes A , Hammock BD , Morisseau C
Ref : Front Vet Sci , 7 :580 , 2020
Abstract : The veterinary pharmacopeia available to treat pain and inflammation is limited in number, target of action and efficacy. Inhibitors of soluble epoxide hydrolase (sEH) are a new class of anti-inflammatory, pro-resolving and analgesic drugs being tested in humans that have demonstrated efficacy in laboratory animals. They block the hydrolysis, and thus, increase endogenous concentrations of analgesic and anti-inflammatory signaling molecules called epoxy-fatty acids. Here, we screened a library of 2,300 inhibitors of the sEH human against partially purified feline, canine and equine hepatic sEH to identify inhibitors that are broadly potent among species. Six very potent sEH inhibitors (IC(50) < 1 nM for each enzyme tested) were identified. Their microsomal stability was then measured in hepatic extracts from cat, dog and horse, as well as their solubility in solvents suitable for the formulation of drugs. The trans-4-{4-[3-(4-trifluoromethoxy-phenyl)-ureido]-cyclohexyloxy}-benzoic acid (t-TUCB, 1,728) appears to be the best compromise between stability and potency across species. Thus, it was selected for further testing in veterinary clinical trials of pain and inflammation in animals.
ESTHER : Shihadih_2020_Front.Vet.Sci_7_580
PubMedSearch : Shihadih_2020_Front.Vet.Sci_7_580
PubMedID: 33005645

Title : The Metabolism of Epoxyeicosatrienoic Acids by Soluble Epoxide Hydrolase Is Protective against the Development of Vascular Calcification - Varennes_2020_Int.J.Mol.Sci_21_
Author(s) : Varennes O , Mentaverri R , Duflot T , Kauffenstein G , Objois T , Lenglet G , Avondo C , Morisseau C , Brazier M , Kamel S , Six I , Bellien J
Ref : Int J Mol Sci , 21 : , 2020
Abstract : This study addressed the hypothesis that soluble epoxide hydrolase (sEH), which metabolizes endothelium-derived epoxyeicosatrienoic acids, plays a role in vascular calcification. The sEH inhibitor trans-4-(4-(3-adamantan-1-yl-ureido)-cyclohexyloxy)-benzoic acid (t-AUCB) potentiated the increase in calcium deposition of rat aortic rings cultured in high-phosphate conditions. This was associated with increased tissue-nonspecific alkaline phosphatase activity and mRNA expression level of the osteochondrogenic marker Runx2. The procalcifying effect of t-AUCB was prevented by mechanical aortic deendothelialization or inhibition of the production and action of epoxyeicosatrienoic acids using the cytochrome P450 inhibitor fluconazole and the antagonist 14,15-epoxyeicosa-5(Z)-enoic acid (14,15-EEZE), respectively. Similarly, exogenous epoxyeicosatrienoic acids potentiated the calcification of rat aortic rings through a protein kinase A (PKA)-dependent mechanism and of human aortic vascular smooth muscle cells when sEH was inhibited by t-AUCB. Finally, a global gene expression profiling analysis revealed that the mRNA expression level of sEH was decreased in human carotid calcified plaques compared to adjacent lesion-free sites and was inversely correlated with Runx2 expression. These results show that sEH hydrolase plays a protective role against vascular calcification by reducing the bioavailability of epoxyeicosatrienoic acids.
ESTHER : Varennes_2020_Int.J.Mol.Sci_21_
PubMedSearch : Varennes_2020_Int.J.Mol.Sci_21_
PubMedID: 32560362

Title : Development of Improved Double Nanobody Sandwich ELISAs for Human Soluble Epoxide Hydrolase Detection in PBMCs of Diabetic and Pre-frontal Cortex of Multiple Sclerosis Patients - Li_2020_Anal.Chem_92_7334
Author(s) : Li D , Morisseau C , McReynolds CB , Duflot T , Bellien J , Nagra RM , Taha AY , Hammock BD
Ref : Analytical Chemistry , 92 :7334 , 2020
Abstract : Nanobodies have been progressively replacing traditional antibodies in various immunological methods. However, the use of nanobodies as capture antibodies is greatly hampered by their poor performance after passive adsorption to polystyrene microplates, and this restricts the full use of double nanobodies in sandwich ELISAs. Herein, using the human soluble epoxide hydrolase (sEH) as a model analyte, we found both the immobilization format and blocking agent have a significant influence on the performance of capture nanobodies immobilized on polystyrene and the subsequent development of double nanobody sandwich ELISAs. We first conducted epitope mapping for pairing nanobodies and then prepared a horseradish peroxidase labeled nanobody using a mild conjugation procedure as detection antibody throughout the work. The resulting sandwich ELISA using capture nanobody (A9, 1.25 mug/mL) after passive adsorption and BSA as blocking agent generated a moderate sensitivity of 0.0164 OD*mL/ng and a LOD of 0.74 ng/mL. However, the introduction of streptavidin as a linker to the capture nanobody at the same working concentration demonstrated a dramatic 16-fold increase in sensitivity (0.262 OD*mL/ng) and 25-fold decrease in the LOD for sEH (0.03 ng/mL). The streptavidin bridged double nanobody ELISA was then applied successfully to tests for recovery, cross-reactivity, and real samples. Meanwhile, we accidentally found that blocking with skim milk could severely damage the performance of the capture nanobody by an order of magnitude, compared to BSA. This work provides guideline to retain the high effectiveness of capture nanobody and thus to further develop double nanobody ELISA for various analytes.
ESTHER : Li_2020_Anal.Chem_92_7334
PubMedSearch : Li_2020_Anal.Chem_92_7334
PubMedID: 32253910

Title : Preparation and evaluation of soluble epoxide hydrolase inhibitors with improved physical properties and potencies for treating diabetic neuropathic pain - Lee_2020_Bioorg.Med.Chem_28_115735
Author(s) : Lee KSS , Ng JC , Yang J , Hwang SH , Morisseau C , Wagner K , Hammock BD
Ref : Bioorganic & Medicinal Chemistry , 28 :115735 , 2020
Abstract : Soluble epoxide hydrolase (sEH), a novel therapeutic target for neuropathic pain, is a largely cytosolic enzyme that degrades epoxy-fatty acids (EpFAs), an important class of lipid signaling molecules. Many inhibitors of sEH have been reported, and to date, the 1,3-disubstituted urea has the highest affinity reported for the sEH among the central pharmacophores evaluated. An earlier somewhat water soluble sEH inhibitor taken to the clinic for blood pressure control had mediocre potency (both affinity and kinetics) and a short in vivo half-life. We undertook a study to overcome these difficulties, but the sEH inhibitors carrying a 1,3-disubstituted urea often suffer poor physical properties that hinder their formulation. In this report, we described new strategies to improve the physical properties of sEH inhibitors with a 1,3-disubstituted urea while maintaining their potency and drug-target residence time (a complementary in vitro parameter) against sEH. To our surprise, we identified two structural modifications that substantially improve the potency and physical properties of sEH inhibitors carrying a 1,3-disubstituted urea pharmacophore. Such improvements will greatly facilitate the movement of sEH inhibitors to the clinic.
ESTHER : Lee_2020_Bioorg.Med.Chem_28_115735
PubMedSearch : Lee_2020_Bioorg.Med.Chem_28_115735
PubMedID: 33007552

Title : Imidazolidine-2,4,5- and pirimidine-2,4,6-triones - New primary pharmacophore for soluble epoxide hydrolase inhibitors with enhanced water solubility - Burmistrov_2020_Bioorg.Med.Chem.Lett_30_126908
Author(s) : Burmistrov V , Morisseau C , D'Yachenko V , Karlov D , Butov GM , Hammock BD
Ref : Bioorganic & Medicinal Chemistry Lett , 30 :126908 , 2020
Abstract : A series of inhibitors of the soluble epoxide hydrolase (sEH) containing imidazolidine-2,4,5-trione or pirimidine-2,4,6-trione has been synthesized. Inhibition potency of the described compounds ranges from 8.4 muM to 0.4 nM. The tested compounds possess higher water solubility than their preceding ureas. Molecular docking indicates new bond between the triones and the active site of sEH that in part explain the observed potency of the new pharmacophores. While less potent than the corresponding ureas, the modifications of urea group reported herein yield compounds with higher water solubility, thus permitting easier formulation.
ESTHER : Burmistrov_2020_Bioorg.Med.Chem.Lett_30_126908
PubMedSearch : Burmistrov_2020_Bioorg.Med.Chem.Lett_30_126908
PubMedID: 31870649

Title : N-Benzyl-linoleamide, a Constituent of Lepidium meyenii (Maca), Is an Orally Bioavailable Soluble Epoxide Hydrolase Inhibitor That Alleviates Inflammatory Pain - Singh_2020_J.Nat.Prod_83_3689
Author(s) : Singh N , Barnych B , Morisseau C , Wagner KM , Wan D , Takeshita A , Pham H , Xu T , Dandekar A , Liu JY , Hammock BD
Ref : Journal of Natural Products , 83 :3689 , 2020
Abstract : Lepidium meyenii (maca), a plant indigenous to the Peruvian Andes, recently has been utilized globally for claimed health or recreational benefits. The search for natural products that inhibit soluble epoxide hydrolase (sEH), with therapeutically relevant potencies and concentrations, led to the present study on bioactive amide secondary metabolites found in L. meyenii, the macamides. Based on known and suspected macamides, 19 possible macamides were synthesized and characterized. The majority of these amides displayed excellent inhibitory potency (IC(50) = 20-300 nM) toward the recombinant mouse, rat, and human sEH. Quantitative analysis of commercial maca products revealed that certain products contain known macamides (1-5, 8-12) at therapeutically relevant total concentrations (<=3.29 mg/g of root), while the inhibitory potency of L. meyenii extracts directly correlates with the sum of concentration/IC(50) ratios of macamides present. Considering both its in vitro efficacy and high abundance in commercial products, N-benzyl-linoleamide (4) was identified as a particularly relevant macamide that can be utilized for in vivo studies. Following oral administration in the rat, compound 4 not only displayed acceptable pharmacokinetic characteristics but effectively reduced lipopolysaccharide-induced inflammatory pain. Inhibition of sEH by macamides provides a plausible biological mechanism of action to account for several beneficial effects previously observed with L. meyenii treatments.
ESTHER : Singh_2020_J.Nat.Prod_83_3689
PubMedSearch : Singh_2020_J.Nat.Prod_83_3689
PubMedID: 33320645

Title : 2-Oxaadamant-1-yl Ureas as Soluble Epoxide Hydrolase Inhibitors: In Vivo Evaluation in a Murine Model of Acute Pancreatitis - Codony_2020_J.Med.Chem_63_9237
Author(s) : Codony S , Pujol E , Pizarro J , Feixas F , Valverde E , Loza MI , Brea JM , Saez E , Oyarzabal J , Pineda-Lucena A , Perez B , Perez C , Rodriguez-Franco MI , Leiva R , Osuna S , Morisseau C , Hammock BD , Vazquez-Carrera M , Vazquez S
Ref : Journal of Medicinal Chemistry , 63 :9237 , 2020
Abstract : In vivo pharmacological inhibition of soluble epoxide hydrolase (sEH) reduces inflammatory diseases, including acute pancreatitis (AP). Adamantyl ureas are very potent sEH inhibitors, but the lipophilicity and metabolism of the adamantane group compromise their overall usefulness. Herein, we report that the replacement of a methylene unit of the adamantane group by an oxygen atom increases the solubility, permeability, and stability of three series of urea-based sEH inhibitors. Most of these oxa-analogues are nanomolar inhibitors of both the human and murine sEH. Molecular dynamics simulations rationalize the molecular basis for their activity and suggest that the presence of the oxygen atom on the adamantane scaffold results in active site rearrangements to establish a weak hydrogen bond. The 2-oxaadamantane 22, which has a good solubility, microsomal stability, and selectivity for sEH, was selected for further in vitro and in vivo studies in models of cerulein-induced AP. Both in prophylactic and treatment studies, 22 diminished the overexpression of inflammatory and endoplasmic reticulum stress markers induced by cerulein and reduced the pancreatic damage.
ESTHER : Codony_2020_J.Med.Chem_63_9237
PubMedSearch : Codony_2020_J.Med.Chem_63_9237
PubMedID: 32787085

Title : Bioisosteric substitution of adamantane with bicyclic lipophilic groups improves water solubility of human soluble epoxide hydrolase inhibitors - Burmistrov_2020_Bioorg.Med.Chem.Lett_30_127430
Author(s) : Burmistrov V , Morisseau C , Karlov D , Pitushkin D , Vernigora A , Rasskazova E , Butov GM , Hammock BD
Ref : Bioorganic & Medicinal Chemistry Lett , 30 :127430 , 2020
Abstract : A series of inhibitors of the soluble epoxide hydrolase (sEH) containing lipophilic groups of natural origin (camphanyl, norcamphanyl, furan-2-yl) were developed. Inhibitory potency ranging from 0.4 nM to 2.16 muM were obtained. While having the same level of inhibitory activity bicyclic ureas are up to 10-fold more soluble than the corresponding ureas containing adamantyl or 4-trifluoromethoxyphenyl substituents. This makes them easier to formulate, more bioavailable and thus more promising as therapeutic sEH inhibitors. Endo/exo-form of compound 2b derived from l-camphor is 14-fold more potent than the corresponding analogue derived from d-camphor (IC(50) = 3.7 nM vs. 50.6 nM) indicating enantiomeric preference.
ESTHER : Burmistrov_2020_Bioorg.Med.Chem.Lett_30_127430
PubMedSearch : Burmistrov_2020_Bioorg.Med.Chem.Lett_30_127430
PubMedID: 32736212

Title : Epoxy Fatty Acids Are Promising Targets for Treatment of Pain, Cardiovascular Disease and Other Indications Characterized by Mitochondrial Dysfunction, Endoplasmic Stress and Inflammation - McReynolds_2020_Adv.Exp.Med.Biol_1274_71
Author(s) : McReynolds C , Morisseau C , Wagner K , Hammock B
Ref : Advances in Experimental Medicine & Biology , 1274 :71 , 2020
Abstract : Bioactive lipid mediators resulting from the metabolism of polyunsaturated fatty acids (PUFA) are controlled by many pathways that regulate the levels of these mediators and maintain homeostasis to prevent disease. PUFA metabolism is driven primarily through three pathways. Two pathways, the cyclooxygenase (COX) and lipoxygenase (LO) enzymatic pathways, form metabolites that are mostly inflammatory, while the third route of metabolism results from the oxidation by the cytochrome P450 enzymes to form hydroxylated PUFA and epoxide metabolites. These epoxygenated fatty acids (EpFA) demonstrate largely anti-inflammatory and beneficial properties, in contrast to the other metabolites formed from the degradation of PUFA. Dysregulation of these systems often leads to chronic disease. Pharmaceutical targets of disease focus on preventing the formation of inflammatory metabolites from the COX and LO pathways, while maintaining the EpFA and increasing their concentration in the body is seen as beneficial to treating and preventing disease. The soluble epoxide hydrolase (sEH) is the major route of metabolism of EpFA. Inhibiting its activity increases concentrations of beneficial EpFA, and often disease states correlate to mutations in the sEH enzyme that increase its activity and decrease the concentrations of EpFA in the body. Recent approaches to increasing EpFA include synthetic mimics that replicate biological activity of EpFA while preventing their metabolism, while other approaches focus on developing small molecule inhibitors to the sEH. Increasing EpFA concentrations in the body has demonstrated multiple beneficial effects in treating many diseases, including inflammatory and painful conditions, cardiovascular disease, neurological and disease of the central nervous system. Demonstration of efficacy in so many disease states can be explained by the fundamental mechanism that EpFA have of maintaining healthy microvasculature and preventing mitochondrial and endoplasmic reticulum stress. While there are no FDA approved methods that target the sEH or other enzymes responsible for metabolizing EpFA, current clinical efforts to test for efficacy by increasing EpFA that include inhibiting the sEH or administration of EpFA mimics that block metabolism are in progress.
ESTHER : McReynolds_2020_Adv.Exp.Med.Biol_1274_71
PubMedSearch : McReynolds_2020_Adv.Exp.Med.Biol_1274_71
PubMedID: 32894508

Title : Development of multitarget inhibitors for the treatment of pain: Design, synthesis, biological evaluation and molecular modeling studies - Wilt_2020_Bioorg.Chem_103_104165
Author(s) : Wilt S , Kodani S , Le TNH , Nguyen L , Vo N , Ly T , Rodriguez M , Hudson PK , Morisseau C , Hammock BD , Pecic S
Ref : Bioorg Chem , 103 :104165 , 2020
Abstract : Multitarget-directed ligands are a promising class of drugs for discovering innovative new therapies for difficult to treat diseases. In this study, we designed dual inhibitors targeting the human fatty acid amide hydrolase (FAAH) enzyme and human soluble epoxide hydrolase (sEH) enzyme. Targeting both of these enzymes concurrently with single target inhibitors synergistically reduces inflammatory and neuropathic pain; thus, dual FAAH/sEH inhibitors are likely to be powerful analgesics. Here, we identified the piperidinyl-sulfonamide moiety as a common pharmacophore and optimized several inhibitors to have excellent inhibition profiles on both targeted enzymes simultaneously. In addition, several inhibitors show good predicted pharmacokinetic properties. These results suggest that this series of inhibitors has the potential to be further developed as new lead candidates and therapeutics in pain management.
ESTHER : Wilt_2020_Bioorg.Chem_103_104165
PubMedSearch : Wilt_2020_Bioorg.Chem_103_104165
PubMedID: 32891856

Title : Development of a Highly Sensitive Enzyme-Linked Immunosorbent Assay for Mouse Soluble Epoxide Hydrolase Detection by Combining a Polyclonal Capture Antibody with a Nanobody Tracer - Li_2020_Anal.Chem_92_11654
Author(s) : Li D , Cui Y , Morisseau C , Wagner KM , Cho YS , Hammock BD
Ref : Analytical Chemistry , 92 :11654 , 2020
Abstract : Enzyme-linked immunosorbent assays (ELISA) for the detection of soluble epoxide hydrolase (sEH), a key enzyme in the metabolism of fatty acids and a biomarker, may increasingly represent an important diagnostic tool. However, there is a lack of ELISAs for mouse sEH quantification, thus resulting in a bottleneck in understanding the pathogenesis of many diseases related to sEH based on mouse models. In this work, nanobodies recognizing mouse sEH were obtained through rebiopanning against mouse sEH in the previous phage display library of human sEH. Later, we developed four ELISAs involving a combination of anti-mouse sEH polyclonal antibodies (pAbs) and nanobodies. It was found that the double antibodies worked as dual filters and had a huge impact on both the sensitivity and selectivity of sandwich immunoassays. The switch from anti-human sEH pAbs to anti-mouse sEH pAbs led to over a 100-fold increase in the sensitivity and a dramatic decrease of the limit of detection to a picogram per milliliter range in format B (pAb/biotin-VHH/streptavidin-poly-horseradish peroxidase). Moreover, we found that the four sandwich ELISAs might demonstrate excellent selectivities to mouse sEH, despite the antibodies alone showing significant cross-reactivity to the matrix, indicating the enhanced selectivity of double antibodies as dual filters. Eventually, for the first time, the ELISA (format B) was successfully used to measure the mouse sEH level in cancer cells with ultralow abundances. The ELISAs proposed here represent a sensitive tool for tracking sEH in various biological processes and also provide deep insights into developing sandwich immunoassays against various targets in terms of both the sensitivity and selectivity.
ESTHER : Li_2020_Anal.Chem_92_11654
PubMedSearch : Li_2020_Anal.Chem_92_11654
PubMedID: 32786492

Title : Editorial: Clinical Paths for Soluble Epoxide Hydrolase Inhibitors -
Author(s) : Imig JD , Morisseau C
Ref : Front Pharmacol , 11 :598858 , 2020
PubMedID: 33071800

Title : Development of potent inhibitors of the human microsomal epoxide hydrolase - Barnych_2020_Eur.J.Med.Chem_193_112206
Author(s) : Barnych B , Singh N , Negrel S , Zhang Y , Magis D , Roux C , Hua X , Ding Z , Morisseau C , Tantillo DJ , Siegel JB , Hammock BD
Ref : Eur Journal of Medicinal Chemistry , 193 :112206 , 2020
Abstract : Microsomal epoxide hydrolase (mEH) hydrolyzes a wide range of epoxide containing molecules. Although involved in the metabolism of xenobiotics, recent studies associate mEH with the onset and development of certain disease conditions. This phenomenon is partially attributed to the significant role mEH plays in hydrolyzing endogenous lipid mediators, suggesting more complex and extensive physiological functions. In order to obtain pharmacological tools to further study the biology and therapeutic potential of this enzyme target, we describe the development of highly potent 2-alkylthio acetamide inhibitors of the human mEH with IC50 values in the low nanomolar range. These are around 2 orders of magnitude more potent than previously obtained primary amine, amide and urea-based mEH inhibitors. Experimental assay results and rationalization of binding through docking calculations of inhibitors to a mEH homology model indicate that an amide connected to an alkyl side chain and a benzyl-thio function as key pharmacophore units.
ESTHER : Barnych_2020_Eur.J.Med.Chem_193_112206
PubMedSearch : Barnych_2020_Eur.J.Med.Chem_193_112206
PubMedID: 32203787
Gene_locus related to this paper: human-EPHX1 , mouse-EPHX1

Title : The molecular structure of an epoxide hydrolase from Trichoderma reesei in complex with urea or amide-based inhibitors - de Oliveira_2019_Int.J.Biol.Macromol_129_653
Author(s) : de Oliveira GS , Adriani PP , Ribeiro JA , Morisseau C , Hammock BD , Dias MVB , Chambergo FS
Ref : Int J Biol Macromol , 129 :653 , 2019
Abstract : Epoxide hydrolases (EHs) are enzymes involved in the metabolism of endogenous and exogenous epoxides, and the development of EH inhibitors has important applications in the medicine. In humans, EH inhibitors are being tested in the treatment of cardiovascular diseases and show potent anti-inflammatory effects. EH inhibitors are also considerate promising molecules against infectious diseases. EHs are functionally very well studied, but only a few members have its three-dimensional structures characterized. Recently, a new EH from the filamentous fungi Trichoderma reseei (TrEH) was reported, and a series of urea or amide-based inhibitors were identified. In this study, we describe the crystallographic structures of TrEH in complex with five different urea or amide-based inhibitors with resolutions ranging from 2.6 to 1.7A. The analysis of these structures reveals the molecular basis of the inhibition of these compounds. We could also observe that these inhibitors occupy the whole extension of the active site groove and only a few conformational changes are involved. Understanding the structural basis EH interactions with different inhibitors might substantially contribute for the study of fungal metabolism and in the development of novel and more efficient antifungal drugs against pathogenic Trichoderma species.
ESTHER : de Oliveira_2019_Int.J.Biol.Macromol_129_653
PubMedSearch : de Oliveira_2019_Int.J.Biol.Macromol_129_653
PubMedID: 30771398
Gene_locus related to this paper: hypjq-g0r7e2

Title : Substrate and inhibitor selectivity, and biological activity of an epoxide hydrolase from Trichoderma reesei - de Oliveira_2019_Mol.Biol.Rep_46_371
Author(s) : de Oliveira GS , Adriani PP , Wu H , Morisseau C , Hammock BD , Chambergo FS
Ref : Mol Biol Rep , 46 :371 , 2019
Abstract : Epoxide hydrolases (EHs) are present in all living organisms and catalyze the hydrolysis of epoxides to the corresponding vicinal diols. EH are involved in the metabolism of endogenous and exogenous epoxides, and thus have application in pharmacology and biotechnology. In this work, we describe the substrates and inhibitors selectivity of an epoxide hydrolase recently cloned from the filamentous fungus Trichoderma reesei QM9414 (TrEH). We also studied the TrEH urea-based inhibitors effects in the fungal growth. TrEH showed high activity on radioative and fluorescent surrogate and natural substrates, especially epoxides from docosahexaenoic acid. Using a fluorescent surrogate substrate, potent inhibitors of TrEH were identified. Interestingly, one of the best compounds inhibit up to 60% of T. reesei growth, indicating an endogenous role for TrEH. These data make TrEH very attractive for future studies about fungal metabolism of fatty acids and possible development of novel drugs for human diseases.
ESTHER : de Oliveira_2019_Mol.Biol.Rep_46_371
PubMedSearch : de Oliveira_2019_Mol.Biol.Rep_46_371
PubMedID: 30426381
Gene_locus related to this paper: hypjq-g0r7e2

Title : In vitro and in vivo Metabolism of a Potent Inhibitor of Soluble Epoxide Hydrolase, 1-(1-Propionylpiperidin-4-yl)-3-(4-(trifluoromethoxy)phenyl)urea - Wan_2019_Front.Pharmacol_10_464
Author(s) : Wan D , Yang J , McReynolds CB , Barnych B , Wagner KM , Morisseau C , Hwang SH , Sun J , Blocher R , Hammock BD
Ref : Front Pharmacol , 10 :464 , 2019
Abstract : 1-(1-Propionylpiperidin-4-yl)-3-(4-(trifluoromethoxy)phenyl)urea (TPPU) is a potent soluble epoxide hydrolase (sEH) inhibitor that is used extensively in research for modulating inflammation and protecting against hypertension, neuropathic pain, and neurodegeneration. Despite its wide use in various animal disease models, the metabolism of TPPU has not been well-studied. A broader understanding of its metabolism is critical for determining contributions of metabolites to the overall safety and effectiveness of TPPU. Herein, we describe the identification of TPPU metabolites using LC-MS/MS strategies. Four metabolites of TPPU (M1-M4) were identified from rat urine by a sensitive and specific LC-MS/MS method with double precursor ion scans. Their structures were further supported by LC-MS/MS comparison with synthesized standards. Metabolites M1 and M2 were formed from hydroxylation on a propionyl group of TPPU; M3 was formed by amide hydrolysis of the 1-propionylpiperdinyl group on TPPU; and M4 was formed by further oxidation of the hydroxylated metabolite M2. Interestingly, the predicted alpha-keto amide metabolite and 4-(trifluoromethoxy)aniline (metabolite from urea cleavage) were not detected by the LC-MRM-MS method. This indicates that if formed, the two potential metabolites represent <0.01% of TPPU metabolism. Species differences in the formation of these four identified metabolites was assessed using liver S9 fractions from dog, monkey, rat, mouse, and human. M1, M2, and M3 were generated in liver S9 fractions from all species, and higher amounts of M3 were generated in monkey S9 fractions compared to other species. In addition, rat and human S9 metabolism showed the highest species similarity based on the quantities of each metabolite. The presence of all four metabolites were confirmed in vivo in rats over 72-h post single oral dose of TPPU. Urine and feces were major routes for TPPU excretion. M1, M4 and parent drug were detected as major substances, and M2 and M3 were minor substances. In blood, M1 accounted for ~9.6% of the total TPPU-related exposure, while metabolites M2, M3, and M4 accounted for <0.4%. All four metabolites were potent inhibitors of human sEH but were less potent than the parent TPPU. In conclusion, TPPU is metabolized via oxidation and amide hydrolysis without apparent breakdown of the urea. The aniline metabolites were not observed either in vitro or in vivo. Our findings increase the confidence in the ability to translate preclinical PK of TPPU in rats to humans and facilitates the potential clinical development of TPPU and other sEH inhibitors.
ESTHER : Wan_2019_Front.Pharmacol_10_464
PubMedSearch : Wan_2019_Front.Pharmacol_10_464
PubMedID: 31143115

Title : Discovery of the First in Vivo Active Inhibitors of the Soluble Epoxide Hydrolase Phosphatase Domain - Kramer_2019_J.Med.Chem_62_8443
Author(s) : Kramer JS , Woltersdorf S , Duflot T , Hiesinger K , Lillich FF , Knoll F , Wittmann SK , Klingler FM , Brunst S , Chaikuad A , Morisseau C , Hammock BD , Buccellati C , Sala A , Rovati GE , Leuillier M , Fraineau S , Rondeaux J , Hernandez-Olmos V , Heering J , Merk D , Pogoryelov D , Steinhilber D , Knapp S , Bellien J , Proschak E
Ref : Journal of Medicinal Chemistry , 62 :8443 , 2019
Abstract : The emerging pharmacological target soluble epoxide hydrolase (sEH) is a bifunctional enzyme exhibiting two different catalytic activities that are located in two distinct domains. Although the physiological role of the C-terminal hydrolase domain is well-investigated, little is known about its phosphatase activity, located in the N-terminal phosphatase domain of sEH (sEH-P). Herein we report the discovery and optimization of the first inhibitor of human and rat sEH-P that is applicable in vivo. X-ray structure analysis of the sEH phosphatase domain complexed with an inhibitor provides insights in the molecular basis of small-molecule sEH-P inhibition and helps to rationalize the structure-activity relationships. 4-(4-(3,4-Dichlorophenyl)-5-phenyloxazol-2-yl)butanoic acid (22b, SWE101) has an excellent pharmacokinetic and pharmacodynamic profile in rats and enables the investigation of the physiological and pathophysiological role of sEH-P in vivo.
ESTHER : Kramer_2019_J.Med.Chem_62_8443
PubMedSearch : Kramer_2019_J.Med.Chem_62_8443
PubMedID: 31436984

Title : Altered bioavailability of epoxyeicosatrienoic acids is associated with conduit artery endothelial dysfunction in type 2 diabetic patients - Duflot_2019_Cardiovasc.Diabetol_18_35
Author(s) : Duflot T , Moreau-Grange L , Roche C , Iacob M , Wils J , Remy-Jouet I , Cailleux AF , Leuillier M , Renet S , Li D , Morisseau C , Lamoureux F , Richard V , Prevost G , Joannides R , Bellien J
Ref : Cardiovasc Diabetol , 18 :35 , 2019
Abstract : BACKGROUND: This pathophysiological study addressed the hypothesis that soluble epoxide hydrolase (sEH), which metabolizes the vasodilator and anti-inflammatory epoxyeicosatrienoic acids (EETs) to dihydroxyeicosatrienoic acids (DHETs), contributes to conduit artery endothelial dysfunction in type 2 diabetes. METHODS AND RESULTS: Radial artery endothelium-dependent flow-mediated dilatation in response to hand skin heating was reduced in essential hypertensive patients (n = 9) and type 2 diabetic subjects with (n = 19) or without hypertension (n = 10) compared to healthy subjects (n = 36), taking into consideration cardiovascular risk factors, flow stimulus and endothelium-independent dilatation to glyceryl trinitrate. Diabetic patients but not non-diabetic hypertensive subjects displayed elevated whole blood reactive oxygen species levels and loss of NO release during heating, assessed by measuring local plasma nitrite variation. Moreover, plasma levels of EET regioisomers increased during heating in healthy subjects, did not change in hypertensive patients and decreased in diabetic patients. Correlation analysis showed in the overall population that the less NO and EETs bioavailability increases during heating, the more flow-mediated dilatation is reduced. The expression and activity of sEH, measured in isolated peripheral blood mononuclear cells, was elevated in diabetic but not hypertensive patients, leading to increased EETs conversion to DHETs. Finally, hyperglycemic and hyperinsulinemic euglycemic clamps induced a decrease in flow-mediated dilatation in healthy subjects and this was associated with an altered EETs release during heating. CONCLUSIONS: These results demonstrate that an increased EETs degradation by sEH and altered NO bioavailability are associated with conduit artery endothelial dysfunction in type 2 diabetic patients independently from their hypertensive status. The hyperinsulinemic and hyperglycemic state in these patients may contribute to these alterations. Trial registration NCT02311075. Registered December 8, 2014.
ESTHER : Duflot_2019_Cardiovasc.Diabetol_18_35
PubMedSearch : Duflot_2019_Cardiovasc.Diabetol_18_35
PubMedID: 30885203

Title : N-Substituted amino acid inhibitors of the phosphatase domain of the soluble epoxide hydrolase - Matsumoto_2019_Biochem.Biophys.Res.Commun_515_248
Author(s) : Matsumoto N , Kataoka M , Hirosaki H , Morisseau C , Hammock BD , Suzuki E , Hasumi K
Ref : Biochemical & Biophysical Research Communications , 515 :248 , 2019
Abstract : The soluble epoxide hydrolase (sEH) is a bifunctional enzyme implicated in the regulation of inflammation. The N-terminal domain harbors a phosphatase activity (N-phos) with an affinity to lipid phosphomonoesters, and the C-terminal domain has an activity to hydrolyze anti-inflammatory lipid epoxides (C-EH). Although many potent inhibitors of C-EH have been discovered, little is known about inhibitors of N-phos. Here, we identify N-substituted amino acids as selective inhibitors of N-phos. Many of the N-substituted amino acids inhibited differently mouse and human N-phos; phenylalanine derivatives are relatively selective for mouse N-phos, whereas tyrosine derivatives are more selective for human N-phos. The best inhibitors, Fmoc-l-Phe(4-CN) (67) and Boc-l-Tyr(Bzl) (23), inhibited mouse and human N-phos competitively with KI in the low micromolar range. These compounds inhibit the N-phos activity 37- (67) and 137-folds (23) more potently than the C-EH. The differences in inhibitor structure activity suggest different active site structure between species, and thus, probably a divergent substrate preference between mouse and human N-phos.
ESTHER : Matsumoto_2019_Biochem.Biophys.Res.Commun_515_248
PubMedSearch : Matsumoto_2019_Biochem.Biophys.Res.Commun_515_248
PubMedID: 31146915

Title : 1-Trifluoromethoxyphenyl-3-(1-propionylpiperidin-4-yl) Urea, a Selective and Potent Dual Inhibitor of Soluble Epoxide Hydrolase and p38 Kinase Intervenes in Alzheimer's Signaling in Human Nerve Cells - Liang_2019_ACS.Chem.Neurosci_10_4018
Author(s) : Liang Z , Zhang B , Xu M , Morisseau C , Hwang SH , Hammock BD , Li QX
Ref : ACS Chem Neurosci , 10 :4018 , 2019
Abstract : Alzheimer's disease (AD) is the most common neurodegenerative disorder. Neuroinflammation is a prevalent pathogenic stress leading to neuronal death in AD. Targeting neuroinflammation to keep neurons alive is an attractive strategy for AD therapy. 1-Trifluoromethoxyphenyl-3-(1-propionylpiperidin-4-yl) urea (TPPU) is a potent inhibitor of soluble epoxide hydrolase (sEH) and can enter into the brain. It has good efficacy on a wide range of chronic inflammatory diseases in preclinical animal models. However, the anti-neuroinflammatory effects and molecular mechanisms of TPPU for potential AD interventions remain elusive. With an aim to develop multitarget therapeutics for neurodegenerative diseases, we screened TPPU against sEH from different mammalian species and a broad panel of human kinases in vitro for potential new targets relevant to neuroinflammation in AD. TPPU inhibits both human sEH and p38beta kinase, two key regulators of inflammation, with nanomolar potencies and distinct selectivity. To further elucidate the molecular mechanisms, differentiated SH-SY5Y human neuroblastoma cells were used as an AD cell model, and we investigated the neuroprotection of TPPU against amyloid oligomers. We found that TPPU effectively prevents neuronal death by mitigating amyloid neurotoxicity, tau hyperphosphorylation, and mitochondrial dysfunction, promoting neurite outgrowth and suppressing activation and nuclear translocation of NF-kappaB for inflammatory responses in human nerve cells. The results indicate that TPPU is a potent and selective dual inhibitor of sEH and p38beta kinase, showing a synergistic action in multiple AD signaling pathways. Our study sheds light upon TPPU and other sEH/p38beta dual inhibitors for potential pharmacological interventions in AD.
ESTHER : Liang_2019_ACS.Chem.Neurosci_10_4018
PubMedSearch : Liang_2019_ACS.Chem.Neurosci_10_4018
PubMedID: 31378059

Title : Phytochemical constituents from Scutellaria baicalensis in soluble epoxide hydrolase inhibition: Kinetics and interaction mechanism merged with simulations - Liu_2019_Int.J.Biol.Macromol_133_1187
Author(s) : Liu ZB , Sun CP , Xu JX , Morisseau C , Hammock BD , Qiu F
Ref : Int J Biol Macromol , 133 :1187 , 2019
Abstract : In our search for soluble epoxide hydrolase (sEH) inhibitors from plants, we found that water extracts of Scutellaria baicalensis Georgi displayed significant inhibitory activity against sEH in vitro. Extracts of S. baicalensiswere separated, resulting in the isolation of thirty compounds (1-30), including six lignins (1-6), sixteen flavones (7-22), and five amides (23-27). Their structures were determined on the basis of(1)H and(13)C NMR and MS spectra. Compounds 1-6 were first reported in the genus Scutellaria. All the isolated compounds were assayed for their inhibitory activities against sEH. Compounds 25-27 showed significant inhibitory activities against sEH with IC50 values of 6.06+/-0.12, 7.83+/-0.52, and 6.32+/-0.31muM, respectively, and compounds 3-6, 12, 18, and 22 displayed moderate inhibitory activities against sEH with IC50 values from 20.82+/-0.78muM to 56.61+/-0.98muM. The inhibition kinetic analysis results indicated that compounds 25-27 were all uncompetitive. Molecular docking studies were performed to get insights into inhibition mechanisms of compounds 25-27 against sEH.
ESTHER : Liu_2019_Int.J.Biol.Macromol_133_1187
PubMedSearch : Liu_2019_Int.J.Biol.Macromol_133_1187
PubMedID: 30980876

Title : Exploring the size of the lipophilic unit of the soluble epoxide hydrolase inhibitors - Codony_2019_Bioorg.Med.Chem_27_115078
Author(s) : Codony S , Valverde E , Leiva R , Brea J , Isabel Loza M , Morisseau C , Hammock BD , Vazquez S
Ref : Bioorganic & Medicinal Chemistry , 27 :115078 , 2019
Abstract : Soluble epoxide hydrolase (sEH) inhibitors are potential drugs for several diseases. Adamantyl ureas are excellent sEH inhibitors but have limited metabolic stability. Herein, we report the effect of replacing the adamantane group by alternative polycyclic hydrocarbons on sEH inhibition, solubility, permeability and metabolic stability. Compounds bearing smaller or larger polycyclic hydrocarbons than adamantane yielded all good inhibition potency of the human sEH (0.4<=IC50<=21.7nM), indicating that sEH is able to accommodate inhibitors of very different size. Human liver microsomal stability of diamantane containing inhibitors is lower than that of their corresponding adamantane counterparts.
ESTHER : Codony_2019_Bioorg.Med.Chem_27_115078
PubMedSearch : Codony_2019_Bioorg.Med.Chem_27_115078
PubMedID: 31488357

Title : Pharmaceutical Effects of Inhibiting the Soluble Epoxide Hydrolase in Canine Osteoarthritis - McReynolds_2019_Front.Pharmacol_10_533
Author(s) : McReynolds CB , Hwang SH , Yang J , Wan D , Wagner K , Morisseau C , Li D , Schmidt WK , Hammock BD
Ref : Front Pharmacol , 10 :533 , 2019
Abstract : Osteoarthritis (OA) is a degenerative joint disease that causes pain and bone deterioration driven by an increase in prostaglandins (PGs) and inflammatory cytokines. Current treatments focus on inhibiting prostaglandin production, a pro-inflammatory lipid metabolite, with NSAID drugs; however, other lipid signaling targets could provide safer and more effective treatment strategies. Epoxides of polyunsaturated fatty acids (PUFAs) are anti-inflammatory lipid mediators that are rapidly metabolized by the soluble epoxide hydrolase (sEH) into corresponding vicinal diols. Interestingly, diol levels are increased in the synovial fluid of humans with OA, warranting further research on the biological role of this lipid pathway in the progression of OA. sEH inhibitors (sEHI) stabilize these biologically active, anti-inflammatory lipid epoxides, resulting in analgesia in both neuropathic, and inflammatory pain conditions. Most experimental studies testing the analgesic effects of sEH inhibitors have used experimental rodent models, which do not completely represent the complex etiology of painful diseases. Here, we tested the efficacy of sEHI in aged dogs with natural arthritis to provide a better representation of the clinical manifestations of pain. Two sEHI were administered orally, once daily for 5 days to dogs with naturally occurring arthritis to assess efficacy and pharmacokinetics. Blinded technicians recorded the behavior of the arthritic dogs based on pre-determined criteria to assess pain and function. After 5 days, EC1728 significantly reduced pain at a dose of 5 mg/kg compared to vehicle controls. Pharmacokinetic evaluation showed concentrations exceeding the enzyme potency in both plasma and synovial fluid. In vitro data showed that epoxyeicosatrienoic acid (EETs), epoxide metabolites of arachidonic acid, decreased inflammatory cytokines, IL-6 and TNF-alpha, and reduced cytotoxicity in canine chondrocytes challenged with IL1beta to simulate an arthritic environment. These results provide the first example of altering lipid epoxides as a therapeutic target for OA potentially acting by protecting chondrocytes from inflammatory induced cytotoxicity. Considering the challenges and high variability of naturally occurring disease in aged dogs, these data provide initial proof of concept justification that inhibiting the sEH is a non-NSAID, non-opioid, disease altering strategy for treating OA, and warrants further investigation.
ESTHER : McReynolds_2019_Front.Pharmacol_10_533
PubMedSearch : McReynolds_2019_Front.Pharmacol_10_533
PubMedID: 31214021

Title : Impact of the acute local inhibition of soluble epoxide hydrolase on diabetic skin microcirculatory dysfunction - Savina_2019_Diab.Vasc.Dis.Res__1479164119860215
Author(s) : Savina Y , Duflot T , Bounoure F , Kotzki S , Thiebaut PA , Serreau PA , Skiba M , Picquenot JM , Cornic M , Morisseau C , Hammock B , Imbert L , Cracowski JL , Richard V , Roustit M , Bellien J
Ref : Diab Vasc Dis Res , :1479164119860215 , 2019
Abstract : The impact of the local inhibition of soluble epoxide hydrolase, which metabolizes vasodilator and anti-inflammatory epoxyeicosanoids, on diabetic skin microvascular dysfunction was assessed. In diabetic db/db mice, basal skin blood flow assessed using laser Doppler imaging was similar to that of control mice, but thermal hyperemia was markedly reduced. At 2 h after the topical administration of an aqueous gel containing the soluble epoxide hydrolase inhibitor trans-4-[4-(3-adamantan-1-yl-ureido)-cyclohexyloxy]-benzoic acid (t-AUCB: 400 mg/L), the peak concentration of t-AUCB was detected in the skin of diabetic mice, which quickly decreased thereafter. In parallel, 2 h after application of t-AUCB treatment, thermal hyperemia was increased compared to the control gel. Quantification of t-AUCB in plasma of treated animals showed no or low systemic diffusion. Furthermore, haematoxylin and eosin histological staining of skin biopsies showed that skin integrity was preserved in t-AUCB-treated mice. Finally, for pig ear skin, a surrogate for human skin, using Franz diffusion cells, we observed a continuous diffusion of t-AUCB from 2 h after application to beyond 24 h. A single topical administration of a soluble epoxide hydrolase inhibitor improves microcirculatory function in the skin of db/db mice and might represent a new therapeutic approach for preventing the development of skin complications in diabetic patients.
ESTHER : Savina_2019_Diab.Vasc.Dis.Res__1479164119860215
PubMedSearch : Savina_2019_Diab.Vasc.Dis.Res__1479164119860215
PubMedID: 31267765

Title : Computer-Aided Selective Optimization of Side Activities of Talinolol - Hiesinger_2019_ACS.Med.Chem.Lett_10_899
Author(s) : Hiesinger K , Kramer JS , Achenbach J , Moser D , Weber J , Wittmann SK , Morisseau C , Angioni C , Geisslinger G , Kahnt AS , Kaiser A , Proschak A , Steinhilber D , Pogoryelov D , Wagner K , Hammock BD , Proschak E
Ref : ACS Med Chem Lett , 10 :899 , 2019
Abstract : Selective optimization of side activities is a valuable source of novel lead structures in drug discovery. In this study, a computer-aided approach was used to deorphanize the pleiotropic cholesterol-lowering effects of the beta-blocker talinolol, which result from the inhibition of the enzyme soluble epoxide hydrolase (sEH). X-ray structure analysis of the sEH in complex with talinolol enables a straightforward optimization of inhibitory potency. The resulting lead structure exhibited in vivo activity in a rat model of diabetic neuropatic pain.
ESTHER : Hiesinger_2019_ACS.Med.Chem.Lett_10_899
PubMedSearch : Hiesinger_2019_ACS.Med.Chem.Lett_10_899
PubMedID: 31223445
Gene_locus related to this paper: human-EPHX2

Title : Role of epoxy-fatty acids and epoxide hydrolases in the pathology of neuro-inflammation - Kodani_2019_Biochimie_159_59
Author(s) : Kodani SD , Morisseau C
Ref : Biochimie , 159 :59 , 2019
Abstract : Neuroinflammation is a physiologic response aimed at protecting the central nervous system during injury. However, unresolved and chronic neuroinflammation can lead to long term damage and eventually neurologic disease including Parkinson's disease, Alzheimer's disease and dementia. Recently, enhancing the concentration of epoxyeicosatrienoic acids (EETs) through blocking their hydrolytic degradation by soluble epoxide hydrolase (sEH) has been applied towards reducing the long-term damage associated with central neurologic insults. Evidence suggests this protective effect is mediated, at least in part, through polarization of microglia to an anti-inflammatory phenotype that blocks the inflammatory actions of prostaglandins and promotes wound repair. This mini-review overviews the epidemiologic basis for using sEH inhibition towards neuroinflammatory disease and pharmacologic studies testing sEH inhibition in several neurologic diseases. Additionally, the combination of sEH inhibition with other eicosanoid signaling pathways is considered as an enhanced approach for developing potent neuroprotectants.
ESTHER : Kodani_2019_Biochimie_159_59
PubMedSearch : Kodani_2019_Biochimie_159_59
PubMedID: 30716359

Title : Soluble Epoxide Hydrolase Inhibitor t-AUCB Promotes Murine Brown Adipogenesis: Role of PPAR Gamma and PPAR Alpha (P21-069-19) - Hildreth_2019_Curr.Dev.Nutr_3_
Author(s) : Hildreth K , Overby H , Kodani S , Morisseau C , Hammock B , Bettaieb A , Zhao L
Ref : Curr Dev Nutr , 3 : , 2019
Abstract : Objectives: Brown adipose tissue has recently emerged as a novel target for obesity treatment and prevention. In contrast to the lipid storing function of white adipocytes, brown adipocytes are responsible for dissipating energy as heat, a process involving uncoupling protein 1 (UCP1). Soluble epoxide hydrolase (sEH) is a cytosolic enzyme that converts epoxy fatty acids (EpFAs) into less active diols. By stabilizing endogenous EpFAs, potent small molecule sEH inhibitors have been shown to be beneficial for many chronic diseases. Several recent papers have reported that sEH inhibitors are able to reduce diet-induced obesity, possibly by upregulating UCP1 expression. In the current study, we sought to study the mechanisms by which sEH inhibitor acts on brown preadipocytes. Methods: The effects of a potent sEH inhibitor, trans-4-[4-(3-adamantan-1-yl-ureido)-cyclohexyloxy]-benzoic acid (t-AUCB), on murine brown adipocyte differentiation were evaluated by lipid accumulation and expression of brown adipocyte marker genes. PPAR alpha and PPAR gamma activation by t-AUCB was measured by their respective transactivation assays. The roles of PPARs were further studied by pharmacological antagonism and knockdown experiments by small RNA interference. Results: We report that sEH expression was increased during murine brown adipocyte differentiation. t-AUCB dose-dependently promoted brown adipocyte differentiation. Moreover, we demonstrate that t-AUCB activated PPAR alpha, but not PPAR gamma. t-AUCB-induced upregulation of thermogenic gene Ucp1 and Pgc1 alpha and the general differentiation marker Fabp4 were significantly attenuated by the antagonist of PPAR alpha, GW6471. In contrast, they were only partially attenuated by the antagonist of PPAR gamma, GW9662, and specific knockdown of PPAR gamma. Conclusions: Our findings suggest that sEH may regulate brown adipogenesis and sEH pharmacological inhibition by t-AUCB promotes brown adipogenesis, possibly through activation of PPAR alpha. Funding Sources: The work is supported by NIH 1R15DK114790-01A1 (to LZ), R00DK100736 (to AB) and R01ES002710 (to BDH).
ESTHER : Hildreth_2019_Curr.Dev.Nutr_3_
PubMedSearch : Hildreth_2019_Curr.Dev.Nutr_3_
PubMedID: 31223951

Title : Identification and optimization of soluble epoxide hydrolase inhibitors with dual potency towards fatty acid amide hydrolase - Kodani_2018_Bioorg.Med.Chem.Lett_28_762
Author(s) : Kodani SD , Bhakta S , Hwang SH , Pakhomova S , Newcomer ME , Morisseau C , Hammock BD
Ref : Bioorganic & Medicinal Chemistry Lett , 28 :762 , 2018
Abstract : Multi-target inhibitors have become increasing popular as a means to leverage the advantages of poly-pharmacology while simplifying drug delivery. Here, we describe dual inhibitors for soluble epoxide hydrolase (sEH) and fatty acid amide hydrolase (FAAH), two targets known to synergize when treating inflammatory and neuropathic pain. The structure activity relationship (SAR) study described herein initially started with t-TUCB (trans-4-[4-(3-trifluoromethoxyphenyl-l-ureido)-cyclohexyloxy]-benzoic acid), a potent sEH inhibitor that was previously shown to weakly inhibit FAAH. Inhibitors with a 6-fold increase of FAAH potency while maintaining high sEH potency were developed by optimization. Interestingly, compared to most FAAH inhibitors that inhibit through time-dependent covalent modification, t-TUCB and related compounds appear to inhibit FAAH through a time-independent, competitive mechanism. These inhibitors are selective for FAAH over other serine hydrolases. In addition, FAAH inhibition by t-TUCB appears to be higher in human FAAH over other species; however, the new dual sEH/FAAH inhibitors have improved cross-species potency. These dual inhibitors may be useful for future studies in understanding the therapeutic application of dual sEH/FAAH inhibition.
ESTHER : Kodani_2018_Bioorg.Med.Chem.Lett_28_762
PubMedSearch : Kodani_2018_Bioorg.Med.Chem.Lett_28_762
PubMedID: 29366648
Gene_locus related to this paper: human-EPHX2

Title : Catalytic activities of mammalian epoxide hydrolases with cis and trans fatty acid epoxides relevant to skin barrier function - Yamanashi_2018_J.Lipid.Res_59_684
Author(s) : Yamanashi H , Boeglin WE , Morisseau C , Davis RW , Sulikowski GA , Hammock BD , Brash AR
Ref : J Lipid Res , 59 :684 , 2018
Abstract : Lipoxygenase (LOX)-catalyzed oxidation of the essential fatty acid, linoleate, represents a vital step in construction of the mammalian epidermal permeability barrier. Analysis of epidermal lipids indicates that linoleate is converted to a trihydroxy derivative by hydrolysis of an epoxy-hydroxy precursor. We evaluated different epoxide hydrolase (EH) enzymes in the hydrolysis of skin-relevant fatty acid epoxides and compared the products to those of acid-catalyzed hydrolysis. In the absence of enzyme, exposure to pH 5 or pH 6 at 37 degreesC for 30 min hydrolyzed fatty acid allylic epoxyalcohols to four trihydroxy products. By contrast, human soluble EH [sEH (EPHX2)] and human or murine epoxide hydrolase-3 [EH3 (EPHX3)] hydrolyzed cis or trans allylic epoxides to single diastereomers, identical to the major isomers detected in epidermis. Microsomal EH [mEH (EPHX1)] was inactive with these substrates. At low substrate concentrations (<10 microM), EPHX2 hydrolyzed 14,15-epoxyeicosatrienoic acid (EET) at twice the rate of the epidermal epoxyalcohol, 9R,10R-trans-epoxy-11E-13R-hydroxy-octadecenoic acid, whereas human or murine EPHX3 hydrolyzed the allylic epoxyalcohol at 31-fold and 39-fold higher rates, respectively. These data implicate the activities of EPHX2 and EPHX3 in production of the linoleate triols detected as end products of the 12R-LOX pathway in the epidermis and implicate their functioning in formation of the mammalian water permeability barrier.
ESTHER : Yamanashi_2018_J.Lipid.Res_59_684
PubMedSearch : Yamanashi_2018_J.Lipid.Res_59_684
PubMedID: 29459481

Title : Orally Available Soluble Epoxide Hydrolase\/Phosphodiesterase 4 Dual Inhibitor Treats Inflammatory Pain - Blocher_2018_J.Med.Chem_61_3541
Author(s) : Blocher R , Wagner KM , Gopireddy RR , Harris TR , Wu H , Barnych B , Hwang SH , Xiang YK , Proschak E , Morisseau C , Hammock BD
Ref : Journal of Medicinal Chemistry , 61 :3541 , 2018
Abstract : Inspired by previously discovered enhanced analgesic efficacy between soluble epoxide hydrolase (sEH) and phosphodiesterase 4 (PDE4) inhibitors, we designed, synthesized and characterized 21 novel sEH/PDE4 dual inhibitors. The best of these displayed good efficacy in in vitro assays. Further pharmacokinetic studies of a subset of four selected compounds led to the identification of a bioavailable dual inhibitor N-(4-methoxy-2-(trifluoromethyl)benzyl)-1-propionylpiperidine-4-carboxamide (MPPA). In a lipopolysaccharide induced inflammatory pain rat model, MPPA rapidly increased in the blood ( Tmax = 30 min; Cmax = 460 nM) after oral administration of 3 mg/kg and reduced inflammatory pain with rapid onset of action correlating with blood levels over a time course of 4 h. Additionally, MPPA does not alter self-motivated exploration of rats with inflammatory pain or the withdrawal latency in control rats.
ESTHER : Blocher_2018_J.Med.Chem_61_3541
PubMedSearch : Blocher_2018_J.Med.Chem_61_3541
PubMedID: 29614224

Title : Adamantyl thioureas as soluble epoxide hydrolase inhibitors - Burmistrov_2018_Bioorg.Med.Chem.Lett_28_2302
Author(s) : Burmistrov V , Morisseau C , Pitushkin D , Karlov D , Fayzullin RR , Butov GM , Hammock BD
Ref : Bioorganic & Medicinal Chemistry Lett , 28 :2302 , 2018
Abstract : A series of inhibitors of the soluble epoxide hydrolase (sEH) containing one or two thiourea groups has been developed. Inhibition potency of the described compounds ranges from 50muM to 7.2nM. 1,7-(Heptamethylene)bis[(adamant-1-yl)thiourea] (6f) was found to be the most potent sEH inhibitor, among the thioureas tested. The inhibitory activity of the thioureas against the human sEH is closer to the value of activity against rat sEH rather than murine sEH. While being less active, thioureas are up to 7-fold more soluble than ureas, which makes them more bioavailable and thus promising as sEH inhibitors.
ESTHER : Burmistrov_2018_Bioorg.Med.Chem.Lett_28_2302
PubMedSearch : Burmistrov_2018_Bioorg.Med.Chem.Lett_28_2302
PubMedID: 29803731

Title : Pharmacokinetics and antinociceptive effects of the soluble epoxide hydrolase inhibitor t-TUCB in horses with experimentally induced radiocarpal synovitis - Guedes_2018_J.Vet.Pharmacol.Ther_41_230
Author(s) : Guedes AGP , Aristizabal F , Sole A , Adedeji A , Brosnan R , Knych H , Yang J , Hwang SH , Morisseau C , Hammock BD
Ref : J Vet Pharmacol Ther , 41 :230 , 2018
Abstract : This study determined the pharmacokinetics, antinociceptive, and anti-inflammatory effects of the soluble epoxide hydrolase (sEH) inhibitor t-TUCB (trans-4-{4-[3-(4-Trifluoromethoxy-phenyl)-ureido]-cyclohexyloxy}-benzoic acid) in horses with lipopolysaccharide (LPS)-induced radiocarpal synovitis. A total of seven adult healthy mares (n = 4-6/treatment) were administered 3 mug LPS into one radiocarpal joint and t-TUCB intravenously (i.v.) at 0 (control), 0.03, 0.1, 0.3, and 1 mg/kg in a blinded, randomized, crossover design with at least 3 weeks washout between. Two investigators independently assigned pain scores (at rest, walk and trot) and lameness scores before and up to 48 hr after t-TUCB/LPS. Responses to touching the joint skin to assess tactile allodynia, plasma, and synovial fluid (SF) t-TUCB concentrations were determined before and up to 48 hr after t-TUCB/LPS. Blood and SF were collected for clinical laboratory evaluations before and up to 48 hr after t-TUCB/LPS. Areas under the curves of pain and lameness scores were calculated and compared between control and treatments. Data were analyzed using repeated measures ANOVA with Dunnett or Bonferroni post-test. p < .05 was considered significant. Data are mean +/- SEM. Compared to control, pain, lameness, and tactile allodynia were significantly lower with 1 mg/kg t-TUCB, but not the other doses. For 0.1, 0.3, and 1 mg/kg t-TUCB treatments, plasma terminal half-lives were 13 +/- 3, 13 +/- 0.5, and 24 +/- 5 hr, and clearances were 68 +/- 15, 48 +/- 5, and 14 +/- 1 ml hr(-1) kg(-1) . The 1 mg/kg t-TUCB reached the SF at high concentrations. There were no important anti-inflammatory effects. In conclusion, sEH inhibition with t-TUCB may provide analgesia in horses with inflammatory joint pain.
ESTHER : Guedes_2018_J.Vet.Pharmacol.Ther_41_230
PubMedSearch : Guedes_2018_J.Vet.Pharmacol.Ther_41_230
PubMedID: 29067696

Title : Design and Potency of Dual Soluble Epoxide Hydrolase\/Fatty Acid Amide Hydrolase Inhibitors - Kodani_2018_ACS.Omega_3_14076
Author(s) : Kodani SD , Wan D , Wagner KM , Hwang SH , Morisseau C , Hammock BD
Ref : ACS Omega , 3 :14076 , 2018
Abstract : Fatty acid amide hydrolase (FAAH) is responsible for regulating concentrations of the endocannabinoid arachidonoyl ethanolamide. Multiple FAAH inhibitors have been developed for clinical trials and have failed to demonstrate efficacy at treating pain, despite promising preclinical data. One approach toward increasing the efficacy of FAAH inhibitors is to concurrently inhibit other targets responsible for regulating pain. Here, we designed dual inhibitors targeting the enzymes FAAH and soluble epoxide hydrolase (sEH), which are targets previously shown to synergize at reducing inflammatory and neuropathic pain. Exploration of the sEH/FAAH inhibitor structure-activity relationship started with PF-750, a FAAH inhibitor (IC50 = 19 nM) that weakly inhibited sEH (IC50 = 640 nM). Potency was optimized resulting in an inhibitor with improved potency on both targets (11, sEH IC50 = 5 nM, FAAH IC50 = 8 nM). This inhibitor demonstrated good target selectivity, pharmacokinetic properties (AUC = 1200 h nM, t 1/2 = 4.9 h in mice), and in vivo target engagement.
ESTHER : Kodani_2018_ACS.Omega_3_14076
PubMedSearch : Kodani_2018_ACS.Omega_3_14076
PubMedID: 30411058

Title : Novel piperidine-derived amide sEH inhibitors as mediators of lipid metabolism with improved stability - Pecic_2018_Prostaglandins.Other.Lipid.Mediat_136_90
Author(s) : Pecic S , Zeki AA , Xu X , Jin GY , Zhang S , Kodani S , Halim M , Morisseau C , Hammock BD , Deng SX
Ref : Prostaglandins Other Lipid Mediat , 136 :90 , 2018
Abstract : We have previously identified and reported several potent piperidine-derived amide inhibitors of the human soluble epoxide hydrolase (sEH) enzyme. The inhibition of this enzyme leads to elevated levels of epoxyeicosatrienoic acids (EETs), which are known to possess anti-inflammatory, vasodilatory, and anti-fibrotic effects. Herein, we report the synthesis of 9 analogs of the lead sEH inhibitor and the follow-up structure-activity relationship and liver microsome stability studies. Our findings show that isosteric modifications that lead to significant alterations in the steric and electronic properties at a specific position in the molecule can reduce the efficacy by up to 75-fold. On the other hand, substituting hydrogen with deuterium produces a notable increase ( approximately 30%) in the molecules' half-lives in both rat and human microsomes, while maintaining sEH inhibition potency. These data highlight the utility of isosteric replacement for improving bioavailability, and the newly-synthesized inhibitor structures may thus, serve as a starting point for preclinical development. Our docking study reveals that in the catalytic pocket of sEH, these analogs are in proximity of the key amino acids involved in hydrolysis of EETs.
ESTHER : Pecic_2018_Prostaglandins.Other.Lipid.Mediat_136_90
PubMedSearch : Pecic_2018_Prostaglandins.Other.Lipid.Mediat_136_90
PubMedID: 29567338

Title : Soluble epoxide hydrolase inhibition alleviates neuropathy in Akita (Ins2 Akita) mice - Wagner_2017_Behav.Brain.Res_326_69
Author(s) : Wagner K , Gilda J , Yang J , Wan D , Morisseau C , Gomes AV , Hammock BD
Ref : Behavioural Brain Research , 326 :69 , 2017
Abstract : The soluble epoxide hydrolase (sEH) is a regulatory enzyme responsible for the metabolism of bioactive lipid epoxides of both omega-6 and omega-3 long chain polyunsaturated fatty acids. These natural epoxides mediate cell signaling in several physiological functions including blocking inflammation, high blood pressure and both inflammatory and neuropathic pain. Inhibition of the sEH maintains the level of endogenous bioactive epoxy-fatty acids (EpFA) and allows them to exert their generally beneficial effects. The Akita (Ins2Akita or Ins2C96Y) mice represent a maturity-onset of diabetes of the young (MODY) model in lean, functionally unimpaired animals, with a sexually dimorphic disease phenotype. This allowed for a test of male and female mice in a battery of functional and nociceptive assays to probe the role of sEH in this system. The results demonstrate that inhibiting the sEH is analgesic in diabetic neuropathy and this occurs in a sexually dimorphic manner. Interestingly, sEH activity is also sexually dimorphic in the Akita model, and moreover correlates with disease status particularly in the hearts of male mice. In addition, in vivo levels of oxidized lipid metabolites also correlate with increased sEH expression and the pathogenesis of disease in this model. Thus, sEH is a target to effectively block diabetic neuropathic pain but also demonstrates a potential role in mitigating the progression of this disease.
ESTHER : Wagner_2017_Behav.Brain.Res_326_69
PubMedSearch : Wagner_2017_Behav.Brain.Res_326_69
PubMedID: 28259677

Title : Pseudomonas aeruginosa sabotages the generation of host proresolving lipid mediators - Flitter_2017_Proc.Natl.Acad.Sci.U.S.A_114_136
Author(s) : Flitter BA , Hvorecny KL , Ono E , Eddens T , Yang J , Kwak DH , Bahl CD , Hampton TH , Morisseau C , Hammock BD , Liu X , Lee JS , Kolls JK , Levy BD , Madden DR , Bomberger JM
Ref : Proc Natl Acad Sci U S A , 114 :136 , 2017
Abstract : Recurrent Pseudomonas aeruginosa infections coupled with robust, damaging neutrophilic inflammation characterize the chronic lung disease cystic fibrosis (CF). The proresolving lipid mediator, 15-epi lipoxin A4 (15-epi LXA4), plays a critical role in limiting neutrophil activation and tissue inflammation, thus promoting the return to tissue homeostasis. Here, we show that a secreted P. aeruginosa epoxide hydrolase, cystic fibrosis transmembrane conductance regulator inhibitory factor (Cif), can disrupt 15-epi LXA4 transcellular biosynthesis and function. In the airway, 15-epi LXA4 production is stimulated by the epithelial-derived eicosanoid 14,15-epoxyeicosatrienoic acid (14,15-EET). Cif sabotages the production of 15-epi LXA4 by rapidly hydrolyzing 14,15-EET into its cognate diol, eliminating a proresolving signal that potently suppresses IL-8-driven neutrophil transepithelial migration in vitro. Retrospective analyses of samples from patients with CF supported the translational relevance of these preclinical findings. Elevated levels of Cif in bronchoalveolar lavage fluid were correlated with lower levels of 15-epi LXA4, increased IL-8 concentrations, and impaired lung function. Together, these findings provide structural, biochemical, and immunological evidence that the bacterial epoxide hydrolase Cif disrupts resolution pathways during bacterial lung infections. The data also suggest that Cif contributes to sustained pulmonary inflammation and associated loss of lung function in patients with CF.
ESTHER : Flitter_2017_Proc.Natl.Acad.Sci.U.S.A_114_136
PubMedSearch : Flitter_2017_Proc.Natl.Acad.Sci.U.S.A_114_136
PubMedID: 27980032
Gene_locus related to this paper: pseae-PA2934

Title : Vascular Endothelial Over-Expression of Human Soluble Epoxide Hydrolase (Tie2-sEH Tr) Attenuates Coronary Reactive Hyperemia in Mice: Role of Oxylipins and omega-Hydroxylases - Hanif_2017_PLoS.One_12_e0169584
Author(s) : Hanif A , Edin ML , Zeldin DC , Morisseau C , Falck JR , Nayeem MA
Ref : PLoS ONE , 12 :e0169584 , 2017
Abstract : Cytochromes P450 metabolize arachidonic acid (AA) into two vasoactive oxylipins with opposing biologic effects: epoxyeicosatrienoic acids (EETs) and omega-(omega)-terminal hydroxyeicosatetraenoic acids (HETEs). EETs have numerous beneficial physiological effects, including vasodilation and protection against ischemia/reperfusion injury, whereas omega-terminal HETEs induce vasoconstriction and vascular dysfunction. We evaluated the effect of these oxylipins on post-ischemic vasodilation known as coronary reactive hyperemia (CRH). CRH prevents the potential harm associated with transient ischemia. The beneficial effects of EETs are reduced after their hydrolysis to dihydroxyeicosatrienoic acids (DHETs) by soluble epoxide hydrolase (sEH). omega-terminal HETEs are formed by omega-hydroxylase family members. The relationship among endothelial over-expression of sEH (Tie2-sEH Tr), the changes in oxylipins it may produce, the pharmacologic inhibition of omega-hydroxylases, activation of PPARgamma, and CRH response to a brief ischemia is not known. We hypothesized that CRH is attenuated in isolated mouse hearts with endothelial sEH over-expression through modulation of oxylipin profiles, whereas both inhibition of omega-hydroxylases and activation of PPARgamma enhance CRH. Compared to WT mice, Tie2-sEH Tr mice had decreased CRH, including repayment volume, repayment duration, and repayment/debt ratio (P < 0.05), whereas inhibition of omega-hydroxylases increased these same CRH parameters in Tie2-sEH Tr mice. Inhibition of sEH with t-AUCB reversed the decreased CRH in Tie2-sEH Tr mice. Endothelial over-expression of sEH significantly changed oxylipin profiles, including decreases in DHETs, mid-chain HETEs, and prostaglandins (P < 0.05). Treatment with rosiglitazone, PPARgamma-agonist, enhanced CRH (P < 0.05) in both Tie2-sEH Tr and wild type (WT) mice. These data demonstrate that endothelial over-expression of sEH (through changing the oxylipin profiles) attenuates CRH, whereas inhibition of omega-hydroxylases and activation of PPARgamma enhance it.
ESTHER : Hanif_2017_PLoS.One_12_e0169584
PubMedSearch : Hanif_2017_PLoS.One_12_e0169584
PubMedID: 28056085
Gene_locus related to this paper: human-EPHX2

Title : Soluble epoxide hydrolase activity and pharmacologic inhibition in horses with chronic severe laminitis - Guedes_2017_Equine.Vet.J_49_345
Author(s) : Guedes A , Galuppo L , Hood D , Hwang SH , Morisseau C , Hammock BD
Ref : Equine Vet J , 49 :345 , 2017
Abstract : BACKGROUND: The roles of soluble epoxide hydrolase and lipid mediators in inflammatory and neuropathic pain could be relevant in laminitis pain management. OBJECTIVES: To determine soluble epoxide hydrolase (sEH) activity in the digital laminae, sEH inhibitor potency in vitro, and efficacy of a sEH inhibitor as an adjunct analgesic therapy in chronic laminitic horses. STUDY DESIGN: In vitro experiments and clinical case series.
METHODS: sEH activity was measured in digital laminae from euthanised healthy and laminitic horses (n = 5-6/group). Potency of 7 synthetic sEH inhibitors was determined in vitro using equine liver cytosol. One of them (t-TUCB; 0.1 mg/kg bwt i.v. every 24 h) was selected based on potency and stability, and used as adjunct therapy in 10 horses with severe chronic laminitis (Obel grades 2, one horse; 3-4, nine horses). Daily assessments of forelimb lifts, pain scores, physiologic and laboratory examinations were performed before (baseline) and during t-TUCB treatment. Data are presented as mean +/- s.d. and 95% confidence intervals (CI).
RESULTS: sEH activity in the digital laminae from laminitic horses (0.9+/-0.6 nmol/min/mg; 95% CI 0.16-1.55 nmol/min/mg) was significantly greater (P = 0.01) than in healthy horses (0.17+/-0.09 nmol/min/mg; CI 0.07-0.26 nmol/min/mg). t-TUCB as an adjunct analgesic up to 10 days (4.3+/-3 days) in laminitic horses was associated with significant reduction in forelimb lifts (36+/-22%; 95% CI 9-64%) and in pain scores (18+/-23%; 95% CI 2-35%) compared with baseline (P = 0.04). One horse developed gas colic and another corneal vascularisation in a blind eye during treatment. No other significant changes were observed. MAIN LIMITATIONS: Absence of control group and evaluator blinding in case series.
CONCLUSIONS: sEH activity is significantly higher in the digital laminae of actively laminitic compared with healthy horses, and use of a potent inhibitor of equine sEH as adjunct analgesic therapy appears to decrease signs of pathologic pain in laminitic horses.
ESTHER : Guedes_2017_Equine.Vet.J_49_345
PubMedSearch : Guedes_2017_Equine.Vet.J_49_345
PubMedID: 27338788

Title : In Vitro Metabolism of Oprozomib, an Oral Proteasome Inhibitor: Role of Epoxide Hydrolases and Cytochrome P450s - Wang_2017_Drug.Metab.Dispos_45_712
Author(s) : Wang Z , Fang Y , Teague J , Wong H , Morisseau C , Hammock BD , Rock DA
Ref : Drug Metabolism & Disposition: The Biological Fate of Chemicals , 45 :712 , 2017
Abstract : Oprozomib is an oral proteasome inhibitor currently under investigation in patients with hematologic malignancies or solid tumors. Oprozomib elicits potent pharmacological actions by forming a covalent bond with the active site N-terminal threonine of the 20S proteasome. Oprozomib has a short half-life across preclinical species and in patients due to systemic clearance via metabolism. Potential for drug-drug interactions (DDIs) could alter the exposure of this potent therapeutic; therefore, a thorough investigation of pathways responsible for metabolism is required. In the present study, the major drug-metabolizing enzyme responsible for oprozomib metabolism was identified in vitro. A diol of oprozomib was found to be the predominant metabolite in human hepatocytes, which formed via direct epoxide hydrolysis. Using recombinant epoxide hydrolases (EHs) and selective EH inhibitors in liver microsomes, microsomal EH (mEH) but not soluble EH (sEH) was found to be responsible for oprozomib diol formation. Coincubation with 2-nonylsulfanyl-propionamide, a selective mEH inhibitor, resulted in a significant decrease in oprozomib disappearance (>80%) with concurrent complete blockage of diol formation in human hepatocytes. On the contrary, a selective sEH inhibitor did not affect oprozomib metabolism. Pretreatment of hepatocytes with the pan-cytochrome P450 (P450) inhibitor 1-aminobenzotriazole resulted in a modest reduction ( approximately 20%) of oprozomib metabolism. These findings indicated that mEH plays a predominant role in oprozomib metabolism. Further studies may be warranted to determine whether drugs that are mEH inhibitors cause clinically significant DDIs with oprozomib. On the other hand, pharmacokinetics of oprozomib is unlikely to be affected by coadministered P450 and sEH inhibitors and/or inducers.
ESTHER : Wang_2017_Drug.Metab.Dispos_45_712
PubMedSearch : Wang_2017_Drug.Metab.Dispos_45_712
PubMedID: 28428366

Title : Probing the orientation of inhibitor and epoxy-eicosatrienoic acid binding in the active site of soluble epoxide hydrolase - Lee_2017_Arch.Biochem.Biophys_613_1
Author(s) : Lee KS , Henriksen NM , Ng CJ , Yang J , Jia W , Morisseau C , Andaya A , Gilson MK , Hammock BD
Ref : Archives of Biochemistry & Biophysics , 613 :1 , 2017
Abstract : Soluble epoxide hydrolase (sEH) is an important therapeutic target of many diseases, such as chronic obstructive pulmonary disease (COPD) and diabetic neuropathic pain. It acts by hydrolyzing and thus regulating specific bioactive long chain polyunsaturated fatty acid epoxides (lcPUFA), like epoxyeicosatrienoic acids (EETs). To better predict which epoxides could be hydrolyzed by sEH, one needs to dissect the important factors and structural requirements that govern the binding of the substrates to sEH. This knowledge allows further exploration of the physiological role played by sEH. Unfortunately, a crystal structure of sEH with a substrate bound has not yet been reported. In this report, new photoaffinity mimics of a sEH inhibitor and EET regioisomers were prepared and used in combination with peptide sequencing and computational modeling, to identify the binding orientation of different regioisomers and enantiomers of EETs into the catalytic cavity of sEH. Results indicate that the stereochemistry of the epoxide plays a crucial role in dictating the binding orientation of the substrate.
ESTHER : Lee_2017_Arch.Biochem.Biophys_613_1
PubMedSearch : Lee_2017_Arch.Biochem.Biophys_613_1
PubMedID: 27983948

Title : Effects of adamantane alterations on soluble epoxide hydrolase inhibition potency, physical properties and metabolic stability - Burmistrov_2017_Bioorg.Chem_76_510
Author(s) : Burmistrov V , Morisseau C , Harris TR , Butov G , Hammock BD
Ref : Bioorg Chem , 76 :510 , 2017
Abstract : Adamantyl groups are widely used in medicinal chemistry. However, metabolism limits their usage. Herein, we report the first systematic study of adamantyl ureas and diureas bearing substituents in bridgehead positions of adamantane and/or spacers between urea groups and adamantane group, and tested their effects on soluble epoxide hydrolase inhibitor potency and metabolic stability. Interestingly, the effect on activity against human and murine sEH varied in opposite ways with each new methyl group introduced into the molecule. Compounds with three methyl substituents in adamantane were very poor inhibitors of murine sEH while still very potent against human sEH. In addition, diureas with terminal groups bigger than sEH catalytic tunnel diameter were still good inhibitors suggesting that the active site of sEH opens to capture the substrate or inhibitor molecule. The introduction of one methyl group leads to 4-fold increase in potency without noticeable loss of metabolic stability compared to the unsubstituted adamantane. However, introduction of two or three methyl groups leads to 8-fold and 98-fold decrease in stability in human liver microsomes for the corresponding compounds.
ESTHER : Burmistrov_2017_Bioorg.Chem_76_510
PubMedSearch : Burmistrov_2017_Bioorg.Chem_76_510
PubMedID: 29310082

Title : Development of amide-based fluorescent probes for selective measurement of carboxylesterase 1 activity in tissue extracts - Kodani_2017_Anal.Biochem_539_81
Author(s) : Kodani SD , Barthelemy M , Kamita SG , Hammock B , Morisseau C
Ref : Analytical Biochemistry , 539 :81 , 2017
Abstract : Carboxylesterases are well known for their role in the metabolism of xenobiotics. However, recent studies have also implicated carboxylesterases in regulating a number of physiological processes including metabolic homeostasis and macrophage development, underlying the need to quantify them individually. Unfortunately, current methods for selectively measuring the catalytic activity of individual carboxylesterases are not sufficiently sensitive to support many biological studies. In order to develop a more sensitive and selective method to measure the activity of human carboxylesterase 1 (hCE1), we generated and tested novel substrates with a fluorescent aminopyridine leaving group. hCE1 showed at least a 10-fold higher preference for the optimized substrate 4-MOMMP than the 13 other esterases tested. Because of the high stability of 4-MOMMP and its hydrolysis product, this substrate can be used to measure esterase activity over extended incubation periods yielding a low picogram (femtomol) limit of detection. This sensitivity is comparable to current ELISA methods; however, the new assay quantifies only the catalytically active enzyme facilitating direct correlation to biological processes. The method described herein may allow hCE1 activity to be used as a biomarker for predicting drug pharmacokinetics, early detection of hepatocellular carcinoma, and other disease states where the activity of hCE1 is altered.
ESTHER : Kodani_2017_Anal.Biochem_539_81
PubMedSearch : Kodani_2017_Anal.Biochem_539_81
PubMedID: 29054529

Title : Active-Site Flexibility and Substrate Specificity in a Bacterial Virulence Factor: Crystallographic Snapshots of an Epoxide Hydrolase - Hvorecny_2017_Structure_25_697
Author(s) : Hvorecny KL , Bahl CD , Kitamura S , Lee KSS , Hammock BD , Morisseau C , Madden DR
Ref : Structure , 25 :697 , 2017
Abstract : Pseudomonas aeruginosa secretes an epoxide hydrolase with catalytic activity that triggers degradation of the cystic fibrosis transmembrane conductance regulator (CFTR) and perturbs other host defense networks. Targets of this CFTR inhibitory factor (Cif) are largely unknown, but include an epoxy-fatty acid. In this class of signaling molecules, chirality can be an important determinant of physiological output and potency. Here we explore the active-site chemistry of this two-step alpha/beta-hydrolase and its implications for an emerging class of virulence enzymes. In combination with hydrolysis data, crystal structures of 15 trapped hydroxyalkyl-enzyme intermediates reveal the stereochemical basis of Cif's substrate specificity, as well as its regioisomeric and enantiomeric preferences. The structures also reveal distinct sets of conformational changes that enable the active site to expand dramatically in two directions, accommodating a surprising array of potential physiological epoxide targets. These new substrates may contribute to Cif's diverse effects in vivo, and thus to the success of P. aeruginosa and other pathogens during infection.
ESTHER : Hvorecny_2017_Structure_25_697
PubMedSearch : Hvorecny_2017_Structure_25_697
PubMedID: 28392259
Gene_locus related to this paper: pseae-PA2934

Title : Occurrence of urea-based soluble epoxide hydrolase inhibitors from the plants in the order Brassicales - Kitamura_2017_PLoS.One_12_e0176571
Author(s) : Kitamura S , Morisseau C , Harris TR , Inceoglu B , Hammock BD
Ref : PLoS ONE , 12 :e0176571 , 2017
Abstract : Recently, dibenzylurea-based potent soluble epoxide hydrolase (sEH) inhibitors were identified in Pentadiplandra brazzeana, a plant in the order Brassicales. In an effort to generalize the concept, we hypothesized that plants that produce benzyl glucosinolates and corresponding isothiocyanates also produce these dibenzylurea derivatives. Our overall aim here was to examine the occurrence of urea derivatives in Brassicales, hoping to find biologically active urea derivatives from plants. First, plants in the order Brassicales were analyzed for the presence of 1, 3-dibenzylurea (compound 1), showing that three additional plants in the order Brassicales produce the urea derivatives. Based on the hypothesis, three dibenzylurea derivatives with sEH inhibitory activity were isolated from maca (Lepidium meyenii) roots. Topical application of one of the identified compounds (compound 3, human sEH IC50 = 222 nM) effectively reduced pain in rat inflammatory pain model, and this compound was bioavailable after oral administration in mice. The biosynthetic pathway of these urea derivatives was investigated using papaya (Carica papaya) seed as a model system. Finally, a small collection of plants from the Brassicales order was grown, collected, extracted and screened for sEH inhibitory activity. Results show that several plants of the Brassicales order could be potential sources of urea-based sEH inhibitors.
ESTHER : Kitamura_2017_PLoS.One_12_e0176571
PubMedSearch : Kitamura_2017_PLoS.One_12_e0176571
PubMedID: 28472063

Title : Reduced carboxylesterase 1 is associated with endothelial injury in methamphetamine-induced pulmonary arterial hypertension - Orcholski_2017_Am.J.Physiol.Lung.Cell.Mol.Physiol_313_L252
Author(s) : Orcholski ME , Khurshudyan A , Shamskhou EA , Yuan K , Chen IY , Kodani SD , Morisseau C , Hammock BD , Hong EM , Alexandrova L , Alastalo TP , Berry G , Zamanian RT , de Jesus Perez VA
Ref : American Journal of Physiology Lung Cell Mol Physiol , 313 :L252 , 2017
Abstract : Pulmonary arterial hypertension is a complication of methamphetamine use (METH-PAH) but the pathogenic mechanisms are unknown. Given that cytochrome P450 2D6 (CYP2D6) and carboxylesterase 1 (CES1) are involved in metabolism of METH and other amphetamine-like compounds, we postulated that loss of function variants could contribute to METH-PAH. While no difference in CYP2D6 expression was seen by lung immunofluorescence, CES1 expression was significantly reduced in endothelium of METH-PAH microvessels. Mass spectrometry analysis showed that healthy pulmonary microvascular endothelial cells (PMVECs) have the capacity to both internalize and metabolize METH. Furthermore, whole exome sequencing data from 18 METH-PAH patients revealed that 94.4% of METH-PAH patients were heterozygous carriers of a single nucleotide variant (SNV, rs115629050) predicted to reduce CES1 activity. PMVECs transfected with this CES1 variant demonstrated significantly higher rates of METH-induced apoptosis. METH exposure results in increased formation of reactive oxygen species (ROS) and a compensatory autophagy response. Compared to healthy cells, CES1-deficient PMVECs lack a robust autophagy response despite higher ROS, which correlates with increased apoptosis. We propose that reduced CES1 expression/activity could promote development of METH-PAH by increasing PMVEC apoptosis and small vessel loss.
ESTHER : Orcholski_2017_Am.J.Physiol.Lung.Cell.Mol.Physiol_313_L252
PubMedSearch : Orcholski_2017_Am.J.Physiol.Lung.Cell.Mol.Physiol_313_L252
PubMedID: 28473326
Gene_locus related to this paper: human-CES1

Title : Effect of Soluble Epoxide Hydrolase on the Modulation of Coronary Reactive Hyperemia: Role of Oxylipins and PPARgamma - Hanif_2016_PLoS.One_11_e0162147
Author(s) : Hanif A , Edin ML , Zeldin DC , Morisseau C , Nayeem MA
Ref : PLoS ONE , 11 :e0162147 , 2016
Abstract : Coronary reactive hyperemia (CRH) is a physiological response to ischemic insult that prevents the potential harm associated with an interruption of blood supply. The relationship between the pharmacologic inhibition of soluble epoxide hydrolase (sEH) and CRH response to a brief ischemia is not known. sEH is involved in the main catabolic pathway of epoxyeicosatrienoic acids (EETs), which are converted into dihydroxyeicosatrienoic acids (DHETs). EETs protect against ischemia/reperfusion injury and have numerous beneficial physiological effects. We hypothesized that inhibition of sEH by t-AUCB enhances CRH in isolated mouse hearts through changing the oxylipin profiles, including an increase in EETs/DHETs ratio. Compared to controls, t-AUCB-treated mice had increased CRH, including repayment volume (RV), repayment duration, and repayment/debt ratio (p < 0.05). Treatment with t-AUCB significantly changed oxylipin profiles, including an increase in EET/DHET ratio, increase in EpOME/DiHOME ratio, increase in the levels of HODEs, decrease in the levels of mid-chain HETEs, and decrease in prostanoids (p < 0.05). Treatment with MS-PPOH (CYP epoxygenase inhibitor) reduced CRH, including RV (p < 0.05). Involvement of PPARgamma in the modulation of CRH was demonstrated using a PPARgamma-antagonist (T0070907) and a PPARgamma-agonist (rosiglitazone). T0070907 reduced CRH (p < 0.05), whereas rosiglitazone enhanced CRH (p < 0.05) in isolated mouse hearts compared to the non-treated. These data demonstrate that sEH inhibition enhances, whereas CYP epoxygenases-inhibition attenuates CRH, PPARgamma mediate CRH downstream of the CYP epoxygenases-EET pathway, and the changes in oxylipin profiles associated with sEH-inhibition collectively contributed to the enhanced CRH.
ESTHER : Hanif_2016_PLoS.One_11_e0162147
PubMedSearch : Hanif_2016_PLoS.One_11_e0162147
PubMedID: 27583776

Title : Screening of soluble epoxide hydrolase inhibitory ingredients from traditional Chinese medicines for anti-inflammatory use - Liu_2016_J.Ethnopharmacol_194_475
Author(s) : Liu JY , Morisseau C , Huang H , Hammock BD
Ref : J Ethnopharmacol , 194 :475 , 2016
Abstract : ETHNOPHARMACOLOGICAL RELEVANCE: Inhibition of soluble epoxide hydrolase (sEH) has been extensively reported to be anti-inflammatory in multiple animal models. Some anti-inflammatory traditional Chinese medicines (TCMs) and a few natural compounds were also found to be inhibitory to sEH in vitro. AIM OF THE STUDY: To determine whether the active intergradient (AI) against sEH of anti-inflammatory TCMs in vitro is anti-inflammatory in vivo and the sEH inhibitory action of the AI contributes to its anti-inflammatory effect in vivo. MATERIALS AND
METHODS: In vitro inhibition assay of the sEH was conducted for the methanol and ethanol extracts of 27 anti-inflammatory TCMs. Two potent extracts were subject to further separation guided by bioassay to afford promising AI against sEH in vitro [Fr.5 of the crude ethanol extract of Rhizoma coptidis (FFCERC)]. Finally, the in vivo anti-inflammatory effect and sEH inhibitory potency of FFCERC was evaluated in a lipopolysacchride (LPS)-challenged murine model of acute systemic inflammation. The inflammatory status was characterized by the inflammatory cytokines TNF-alpha and interleukin-6 (IL-6) and sEH inhibitory function was evaluated by the plasma levels of epoxyeicosantrienoic acids (EETs) and dihydroxyeicosatrienoic acids (DHETs), which are the sEH mediated substrates and products, respectively.
RESULTS: At the concentration of 25microg/mL, the crude ethanol extracts of 6 TCMs including Herba Asari, Radix Polygalae, Fructus Amomi, Radix Astragali, Radix Scutellariae, and Rhizoma Coptidis were potent against sEH. The crude extracts of Herba Asari and Rhizoma Coptidis were selected for further separation to afford FFCERC as the most promising AI for in vivo evaluation. Oral administration of FFCERC attenuated the significant increase in TNF-alpha and IL-6 caused by LPS challenge in a dose-dependent manner. In parallel, oral administration of FFCERC shifted the changes in plasma levels of EETs and DHETs caused by LPS-challenge like a synthetic sEH inhibitor.
CONCLUSIONS: A sEH inhibitory AI from Rhizoma Coptidis is anti-inflammatory and the inhibition of sEH contributes to this biological effect, indicating that sEH may be at least one of multiple therapeutic targets for relevant TCMs.
ESTHER : Liu_2016_J.Ethnopharmacol_194_475
PubMedSearch : Liu_2016_J.Ethnopharmacol_194_475
PubMedID: 27702689

Title : Visualizing the Mechanism of Epoxide Hydrolysis by the Bacterial Virulence Enzyme Cif - Bahl_2016_Biochemistry_55_788
Author(s) : Bahl CD , Hvorecny KL , Morisseau C , Gerber SA , Madden DR
Ref : Biochemistry , 55 :788 , 2016
Abstract : The CFTR inhibitory factor (Cif) is an epoxide hydrolase (EH) virulence factor secreted by the bacterium Pseudomonas aeruginosa. Sequence alignments reveal a pattern of Cif-like substitutions that proved to be characteristic of a new subfamily of bacterial EHs. At the same time, crystallographic and mutagenetic data suggest that EH activity is required for virulence and that Cif's active site remains generally compatible with a canonical two-step EH mechanism. A hallmark of this mechanism is the formation of a covalent hydroxyalkyl-enzyme intermediate by nucleophilic attack. In several well-studied EHs, this intermediate has been captured at near stoichiometric levels, presumably reflecting rate-limiting hydrolysis. Here we show by mass spectrometry that only minimal levels of the expected intermediate can be trapped with WT Cif. In contrast, substantial amounts of intermediate are recovered from an active-site mutant (Cif-E153Q) that selectively targets the second, hydrolytic release step. Utilizing Cif-E153Q and a previously reported nucleophile mutant (Cif-D129S), we then captured Cif in the substrate-bound, hydroxyalkyl-intermediate, and product-bound states for 1,2-epoxyhexane, yielding the first crystallographic snapshots of an EH at these key stages along the reaction coordinate. Taken together, our data illuminate the proposed two-step hydrolytic mechanism of a new class of bacterial virulence factor. They also suggest that the failure of WT Cif to accumulate a covalent hydroxyalkyl-enzyme intermediate reflects an active-site chemistry in which hydrolysis is no longer the rate-limiting step, a noncanonical kinetic regime that may explain similar observations with a number of other EHs.
ESTHER : Bahl_2016_Biochemistry_55_788
PubMedSearch : Bahl_2016_Biochemistry_55_788
PubMedID: 26752215
Gene_locus related to this paper: pseae-PA2934

Title : Anti-Ulcer Efficacy of Soluble Epoxide Hydrolase Inhibitor TPPU on Diclofenac-Induced Intestinal Ulcers - Goswami_2016_J.Pharmacol.Exp.Ther_357_529
Author(s) : Goswami SK , Wan D , Yang J , Trindade da Silva CA , Morisseau C , Kodani SD , Yang GY , Inceoglu B , Hammock BD
Ref : Journal of Pharmacology & Experimental Therapeutics , 357 :529 , 2016
Abstract : Proton pump inhibitors such as omeprazole (OME) reduce the severity of gastrointestinal (GI) ulcers induced by nonsteroidal anti-inflammatory drugs (NSAIDs) but can also increase the chance of dysbiosis. The aim of this study was to test the hypothesis that preventive use of a soluble epoxide hydrolase inhibitor (sEHI) such as TPPU can decrease NSAID-induced ulcers by increasing anti-inflammatory epoxyeicosatrienoic acids (EETs). Dose- [10, 30, and 100 mg/kg, by mouth (PO)] and time-dependent (6 and 18 hours) ulcerative effects of diclofenac sodium (DCF, an NSAID) were studied in the small intestine of Swiss Webster mice. Dose-dependent effects of TPPU (0.001-0.1 mg/kg per day for 7 days, in drinking water) were evaluated in DCF-induced intestinal toxicity and compared with OME (20 mg/kg, PO). In addition, the effect of treatment was studied on levels of Hb in blood, EETs in plasma, inflammatory markers such as myeloperoxidase (MPO) in intestinal tissue homogenates, and tissue necrosis factor-alpha (TNF-alpha) in serum. DCF dose dependently induced ulcers that were associated with both a significant (P < 0.05) loss of Hb and an increase in the level of MPO and TNF-alpha, with severity of ulceration highest at 18 hours. Pretreatment with TPPU dose dependently prevented ulcer formation by DCF, increased the levels of epoxy fatty acids, including EETs, and TPPU's efficacy was comparable to OME. TPPU significantly (P < 0.05) reversed the effect of DCF on the level of Hb, MPO, and TNF-alpha Thus sEHI might be useful in the management of NSAID-induced ulcers.
ESTHER : Goswami_2016_J.Pharmacol.Exp.Ther_357_529
PubMedSearch : Goswami_2016_J.Pharmacol.Exp.Ther_357_529
PubMedID: 26989141

Title : Ingestion of the epoxide hydrolase inhibitor AUDA modulates immune responses of the mosquito, Culex quinquefasciatus during blood feeding - Xu_2016_Insect.Biochem.Mol.Biol_76_62
Author(s) : Xu J , Morisseau C , Yang J , Lee KS , Kamita SG , Hammock BD
Ref : Insect Biochemistry & Molecular Biology , 76 :62 , 2016
Abstract : Epoxide hydrolases (EHs) are enzymes that play roles in metabolizing xenobiotic epoxides from the environment, and in regulating lipid signaling molecules, such as juvenile hormones in insects and epoxy fatty acids in mammals. In this study we fed mosquitoes with an epoxide hydrolase inhibitor AUDA during artificial blood feeding, and we found the inhibitor increased the concentration of epoxy fatty acids in the midgut of female mosquitoes. We also observed ingestion of AUDA triggered early expression of defensin A, cecropin A and cecropin B2 at 6 h after blood feeding. The expression of cecropin B1 and gambicin were not changed more than two fold compared to controls. The changes in gene expression were transient possibly because more than 99% of the inhibitor was metabolized or excreted at 42 h after being ingested. The ingestion of AUDA also affected the growth of bacteria colonizing in the midgut, but did not affect mosquito longevity, fecundity and fertility in our laboratory conditions. When spiked into the blood, EpOMEs and DiHOMEs were as effective as the inhibitor AUDA in reducing the bacterial load in the midgut, while EETs rescued the effects of AUDA. Our data suggest that epoxy fatty acids from host blood are immune response regulators metabolized by epoxide hydrolases in the midgut of female mosquitoes, inhibition of which causes transient changes in immune responses, and affects growth of microbes in the midgut.
ESTHER : Xu_2016_Insect.Biochem.Mol.Biol_76_62
PubMedSearch : Xu_2016_Insect.Biochem.Mol.Biol_76_62
PubMedID: 27369469

Title : Gene deficiency and pharmacological inhibition of soluble epoxide hydrolase confers resilience to repeated social defeat stress - Ren_2016_Proc.Natl.Acad.Sci.U.S.A_113_E1944
Author(s) : Ren Q , Ma M , Ishima T , Morisseau C , Yang J , Wagner KM , Zhang JC , Yang C , Yao W , Dong C , Han M , Hammock BD , Hashimoto K
Ref : Proc Natl Acad Sci U S A , 113 :E1944 , 2016
Abstract : Depression is a severe and chronic psychiatric disease, affecting 350 million subjects worldwide. Although multiple antidepressants have been used in the treatment of depressive symptoms, their beneficial effects are limited. The soluble epoxide hydrolase (sEH) plays a key role in the inflammation that is involved in depression. Thus, we examined here the role of sEH in depression. In both inflammation and social defeat stress models of depression, a potent sEH inhibitor, TPPU, displayed rapid antidepressant effects. Expression of sEH protein in the brain from chronically stressed (susceptible) mice was higher than of control mice. Furthermore, expression of sEH protein in postmortem brain samples of patients with psychiatric diseases, including depression, bipolar disorder, and schizophrenia, was higher than controls. This finding suggests that increased sEH levels might be involved in the pathogenesis of certain psychiatric diseases. In support of this hypothesis, pretreatment with TPPU prevented the onset of depression-like behaviors after inflammation or repeated social defeat stress. Moreover, sEH KO mice did not show depression-like behavior after repeated social defeat stress, suggesting stress resilience. The sEH KO mice showed increased brain-derived neurotrophic factor (BDNF) and phosphorylation of its receptor TrkB in the prefrontal cortex, hippocampus, but not nucleus accumbens, suggesting that increased BDNF-TrkB signaling in the prefrontal cortex and hippocampus confer stress resilience. All of these findings suggest that sEH plays a key role in the pathophysiology of depression, and that epoxy fatty acids, their mimics, as well as sEH inhibitors could be potential therapeutic or prophylactic drugs for depression.
ESTHER : Ren_2016_Proc.Natl.Acad.Sci.U.S.A_113_E1944
PubMedSearch : Ren_2016_Proc.Natl.Acad.Sci.U.S.A_113_E1944
PubMedID: 26976569
Gene_locus related to this paper: mouse-hyes

Title : Parabens inhibit fatty acid amide hydrolase: A potential role in paraben-enhanced 3T3-L1 adipocyte differentiation - Kodani_2016_Toxicol.Lett_262_92
Author(s) : Kodani SD , Overby HB , Morisseau C , Chen J , Zhao L , Hammock BD
Ref : Toxicol Lett , 262 :92 , 2016
Abstract : Parabens are a class of small molecules that are regularly used as preservatives in a variety of personal care products. Several parabens, including butylparaben and benzylparaben, have been found to interfere with endocrine signaling and to stimulate adipocyte differentiation. We hypothesized these biological effects could be due to interference with the endocannabinoid system and identified fatty acid amide hydrolase (FAAH) as the direct molecular target of parabens. FAAH inhibition by parabens yields mixed-type and time-independent kinetics. Additionally, structure activity relationships indicate FAAH inhibition is selective for the paraben class of compounds and the more hydrophobic parabens have higher potency. Parabens enhanced 3T3-L1 adipocyte differentiation in a dose dependent fashion, different from two other FAAH inhibitors URB597 and PF622. Moreover, parabens, URB597 and PF622 all failed to enhance AEA-induced differentiation. Furthermore, rimonabant, a cannabinoid receptor 1 (CB1)-selective antagonist, did not attenuate paraben-induced adipocyte differentiation. Thus, adipogenesis mediated by parabens likely occurs through modulation of endocannabinoids, but cell differentiation is independent of direct activation of CB1 by endocannabinoids.
ESTHER : Kodani_2016_Toxicol.Lett_262_92
PubMedSearch : Kodani_2016_Toxicol.Lett_262_92
PubMedID: 27659731

Title : Dysregulation of soluble epoxide hydrolase and lipidomic profiles in anorexia nervosa - Shih_2016_Mol.Psychiatry_21_537
Author(s) : Shih PB , Yang J , Morisseau C , German JB , Zeeland AA , Armando AM , Quehenberger O , Bergen AW , Magistretti P , Berrettini W , Halmi KA , Schork N , Hammock BD , Kaye W
Ref : Mol Psychiatry , 21 :537 , 2016
Abstract : Individuals with anorexia nervosa (AN) restrict eating and become emaciated. They tend to have an aversion to foods rich in fat. Because epoxide hydrolase 2 (EPHX2) was identified as a novel AN susceptibility gene, and because its protein product, soluble epoxide hydrolase (sEH), converts bioactive epoxides of polyunsaturated fatty acid (PUFA) to the corresponding diols, lipidomic and metabolomic targets of EPHX2 were assessed to evaluate the biological functions of EPHX2 and their role in AN. Epoxide substrates of sEH and associated oxylipins were measured in ill AN, recovered AN and gender- and race-matched controls. PUFA and oxylipin markers were tested as potential biomarkers for AN. Oxylipin ratios were calculated as proxy markers of in vivo sEH activity. Several free- and total PUFAs were associated with AN diagnosis and with AN recovery. AN displayed elevated n-3 PUFAs and may differ from controls in PUFA elongation and desaturation processes. Cytochrome P450 pathway oxylipins from arachidonic acid, linoleic acid, alpha-linolenic acid and docosahexaenoic acid PUFAs are associated with AN diagnosis. The diol:epoxide ratios suggest the sEH activity is higher in AN compared with controls. Multivariate analysis illustrates normalization of lipidomic profiles in recovered ANs. EPHX2 influences AN risk through in vivo interaction with dietary PUFAs. PUFA composition and concentrations as well as sEH activity may contribute to the pathogenesis and prognosis of AN. Our data support the involvement of EPHX2-associated lipidomic and oxylipin dysregulations in AN, and reveal their potential as biomarkers to assess responsiveness to future intervention or treatment.
ESTHER : Shih_2016_Mol.Psychiatry_21_537
PubMedSearch : Shih_2016_Mol.Psychiatry_21_537
PubMedID: 25824304
Gene_locus related to this paper: human-EPHX2

Title : Deletion of soluble epoxide hydrolase enhances coronary reactive hyperemia in isolated mouse heart: role of oxylipins and PPARgamma - Hanif_2016_Am.J.Physiol.Regul.Integr.Comp.Physiol_311_R676
Author(s) : Hanif A , Edin ML , Zeldin DC , Morisseau C , Nayeem MA
Ref : American Journal of Physiology Regul Integr Comp Physiol , 311 :R676 , 2016
Abstract : The relationship between soluble epoxide hydrolase (sEH) and coronary reactive hyperemia (CRH) response to a brief ischemic insult is not known. Epoxyeicosatrienoic acids (EETs) exert cardioprotective effects in ischemia/reperfusion injury. sEH converts EETs into dihydroxyeicosatrienoic-acids (DHETs). Therefore, we hypothesized that knocking out sEH enhances CRH through modulation of oxylipin profiles, including an increase in EET/DHET ratio. Compared with sEH+/+, sEH-/- mice showed enhanced CRH, including greater repayment volume (RV; 28% higher, P < 0.001) and repayment/debt ratio (32% higher, P < 0.001). Oxylipins from the heart perfusates were analyzed by LC-MS/MS. The 14,15-EET/14,15-DHET ratio was 3.7-fold higher at baseline (P < 0.001) and 5.6-fold higher post-ischemia (P < 0.001) in sEH-/- compared with sEH+/+ mice. Likewise, the baseline 9,10- and 12,13-EpOME/DiHOME ratios were 3.2-fold (P < 0.01) and 3.7-fold (P < 0.001) higher, respectively in sEH-/- compared with sEH+/+ mice. 13-HODE was also significantly increased at baseline by 71% (P < 0.01) in sEH-/- vs. sEH+/+ mice. Levels of 5-, 11-, 12-, and 15-hydroxyeicosatetraenoic acids were not significantly different between the two strains (P > 0.05), but were decreased postischemia in both groups (P = 0.02, P = 0.04, P = 0.05, P = 0.03, respectively). Modulation of CRH by peroxisome proliferator-activated receptor gamma (PPARgamma) was demonstrated using a PPARgamma-antagonist (T0070907), which reduced repayment volume by 25% in sEH+/+ (P < 0.001) and 33% in sEH-/- mice (P < 0.01), and a PPARgamma-agonist (rosiglitazone), which increased repayment volume by 37% in both sEH+/+ (P = 0.04) and sEH-/- mice (P = 0.04). l-NAME attenuated CRH in both sEH-/- and sEH+/+ These data demonstrate that genetic deletion of sEH resulted in an altered oxylipin profile, which may have led to an enhanced CRH response.
ESTHER : Hanif_2016_Am.J.Physiol.Regul.Integr.Comp.Physiol_311_R676
PubMedSearch : Hanif_2016_Am.J.Physiol.Regul.Integr.Comp.Physiol_311_R676
PubMedID: 27488890
Gene_locus related to this paper: human-EPHX2

Title : Rational Design of Potent and Selective Inhibitors of an Epoxide Hydrolase Virulence Factor from Pseudomonas aeruginosa - Kitamura_2016_J.Med.Chem_59_4790
Author(s) : Kitamura S , Hvorecny KL , Niu J , Hammock BD , Madden DR , Morisseau C
Ref : Journal of Medicinal Chemistry , 59 :4790 , 2016
Abstract : The virulence factor cystic fibrosis transmembrane conductance regulator (CFTR) inhibitory factor (Cif) is secreted by Pseudomonas aeruginosa and is the founding member of a distinct class of epoxide hydrolases (EHs) that triggers the catalysis-dependent degradation of the CFTR. We describe here the development of a series of potent and selective Cif inhibitors by structure-based drug design. Initial screening revealed 1a (KB2115), a thyroid hormone analog, as a lead compound with low micromolar potency. Structural requirements for potency were systematically probed, and interactions between Cif and 1a were characterized by X-ray crystallography. On the basis of these data, new compounds were designed to yield additional hydrogen bonding with residues of the Cif active site. From this effort, three compounds were identified that are 10-fold more potent toward Cif than our first-generation inhibitors and have no detectable thyroid hormone-like activity. These inhibitors will be useful tools to study the pathological role of Cif and have the potential for clinical application.
ESTHER : Kitamura_2016_J.Med.Chem_59_4790
PubMedSearch : Kitamura_2016_J.Med.Chem_59_4790
PubMedID: 27120257
Gene_locus related to this paper: pseae-PA2934

Title : Heavy chain single-domain antibodies to detect native human soluble epoxide hydrolase - Cui_2015_Anal.Bioanal.Chem_407_7275
Author(s) : Cui Y , Li D , Morisseau C , Dong JX , Yang J , Wan D , Rossotti MA , Gee SJ , Gonzalez-Sapienza GG , Hammock BD
Ref : Anal Bioanal Chem , 407 :7275 , 2015
Abstract : The soluble epoxide hydrolase (sEH) is a potential pharmacological target for treating hypertension, vascular inflammation, pain, cancer, and other diseases. However, there is not a simple, inexpensive, and reliable method to estimate levels of active sEH in tissues. Toward developing such an assay, a polyclonal variable domain of heavy chain antibody (VHH) sandwich immunoassay was developed. Ten VHHs, which are highly selective for native human sEH, were isolated from a phage-displayed library. The ten VHHs have no significant cross-reactivity with human microsomal epoxide hydrolase, rat and mouse sEH, and denatured human sEH. There is a high correlation between protein levels of the sEH determined by the enzyme-linked immunosorbent assay (ELISA) and the catalytic activity of the enzyme in S9 fractions of human tissues (liver, kidney, and lung). The VHH-based ELISA appears to be a new reliable method for monitoring the sEH and may be useful as a diagnostic tool for diseases influenced by sEH. This study also demonstrates the broad utility of VHH in biochemical and pharmacological research.
ESTHER : Cui_2015_Anal.Bioanal.Chem_407_7275
PubMedSearch : Cui_2015_Anal.Bioanal.Chem_407_7275
PubMedID: 26229025

Title : 1,3-Disubstituted and 1,3,3-trisubstituted adamantyl-ureas with isoxazole as soluble epoxide hydrolase inhibitors - Burmistrov_2015_Bioorg.Med.Chem.Lett_25_5514
Author(s) : Burmistrov V , Morisseau C , Danilov D , Harris TR , Dalinger I , Vatsadze I , Shkineva T , Butov GM , Hammock BD
Ref : Bioorganic & Medicinal Chemistry Lett , 25 :5514 , 2015
Abstract : Adamantyl ureas are good soluble epoxide hydrolase (sEH) inhibitors; however they have limited solubility and rapid metabolism, thus limiting their usefulness in some therapeutic indications. Herein, we test the hypothesis that nodal substitution on the adamantane will help solubilize and stabilize the compounds. A series of compounds containing adamantane derivatives and isoxazole functional groups were developed. Overall, the presence of methyl on the nodal positions of adamantane yields higher water solubility than previously reported urea-based sEH inhibitors while maintaining high inhibition potency. However, it did not improve microsomal stability.
ESTHER : Burmistrov_2015_Bioorg.Med.Chem.Lett_25_5514
PubMedSearch : Burmistrov_2015_Bioorg.Med.Chem.Lett_25_5514
PubMedID: 26520661

Title : In vitro and in vivo metabolism of N-adamantyl substituted urea-based soluble epoxide hydrolase inhibitors - Liu_2015_Biochem.Pharmacol_98_718
Author(s) : Liu JY , Tsai HJ , Morisseau C , Lango J , Hwang SH , Watanabe T , Kim IH , Hammock BD
Ref : Biochemical Pharmacology , 98 :718 , 2015
Abstract : N,N'-disubstituted urea-based soluble epoxide hydrolase (sEH) inhibitors are promising therapeutics for hypertension, inflammation, and pain in multiple animal models. The drug absorption and pharmacological efficacy of these inhibitors have been reported extensively. However, the drug metabolism of these inhibitors is not well described. Here we reported the metabolic profile and associated biochemical studies of an N-adamantyl urea-based sEH inhibitor 1-adamantan-1-yl-3-(5-(2-(2-ethoxyethoxy)ethoxy)pentyl)urea (AEPU) in vitro and in vivo. The metabolites of AEPU were identified by interpretation of liquid chromatography-mass spectrometry (LC-MS), liquid chromatography-tandem mass spectrometry (LC-MS/MS) and/or NMR. In vitro, AEPU had three major positions for phase I metabolism including oxidations on the adamantyl moiety, urea nitrogen atoms, and cleavage of the polyethylene glycol chain. In a rodent model, the metabolites from the hydroxylation on the adamantyl group and nitrogen atom were existed in blood while the metabolites from cleavage of polyethylene glycol chain were not found in urine. The major metabolite found in rodent urine was 3-(3-adamantyl-ureido)-propanoic acid, a presumably from cleavage and oxidation of the polyethylene glycol moiety. All the metabolites found were active but less potent than AEPU at inhibiting human sEH. Furthermore, cytochrome P450 (CYP) 3A4 was found to be a major enzyme mediating AEPU metabolism. In conclusion, the metabolism of AEPU resulted from oxidation by CYP could be shared with other N-adamantyl-urea-based compounds. These findings suggest possible therapeutic roles for AEPU and new strategies for drug design in this series of possible drugs.
ESTHER : Liu_2015_Biochem.Pharmacol_98_718
PubMedSearch : Liu_2015_Biochem.Pharmacol_98_718
PubMedID: 26494425

Title : Inhibition of soluble epoxide hydrolase in mice promotes reverse cholesterol transport and regression of atherosclerosis - Shen_2015_Atherosclerosis_239_557
Author(s) : Shen L , Peng H , Peng R , Fan Q , Zhao S , Xu D , Morisseau C , Chiamvimonvat N , Hammock BD
Ref : Atherosclerosis , 239 :557 , 2015
Abstract : Adipose tissue is the body largest free cholesterol reservoir and abundantly expresses ATP binding cassette transporter A1 (ABCA1), which maintains plasma high-density lipoprotein (HDL) levels. HDLs have a protective role in atherosclerosis by mediating reverse cholesterol transport (RCT). Soluble epoxide hydrolase (sEH) is a cytosolic enzyme whose inhibition has various beneficial effects on cardiovascular disease. The sEH is highly expressed in adipocytes, and it converts epoxyeicosatrienoic acids (EETs) into less bioactive dihydroxyeicosatrienoic acids. We previously showed that increasing EETs levels with a sEH inhibitor (sEHI) (t-AUCB) resulted in elevated ABCA1 expression and promoted ABCA1-mediated cholesterol efflux from 3T3-L1 adipocytes. The present study investigates the impacts of t-AUCB in mice deficient for the low density lipoprotein (LDL) receptor (Ldlr(-/-) mice) with established atherosclerotic plaques. The sEH inhibitor delivered in vivo for 4 weeks decreased the activity of sEH in adipose tissue, enhanced ABCA1 expression and cholesterol efflux from adipose depots, and consequently increased HDL levels. Furthermore, t-AUCB enhanced RCT to the plasma, liver, bile and feces. It also showed the reduction of plasma LDL-C levels. Consistently, t-AUCB-treated mice showed reductions in the size of atherosclerotic plaques. These studies establish that raising adipose ABCA1 expression, cholesterol efflux, and plasma HDL levels with t-AUCB treatment promotes RCT, decreasing LDL-C and atherosclerosis regression, suggesting that sEH inhibition may be a promising strategy to treat atherosclerotic vascular disease.
ESTHER : Shen_2015_Atherosclerosis_239_557
PubMedSearch : Shen_2015_Atherosclerosis_239_557
PubMedID: 25733327

Title : Oral treatment of rodents with soluble epoxide hydrolase inhibitor 1-(1-propanoylpiperidin-4-yl)-3-[4-(trifluoromethoxy)phenyl]urea (TPPU): Resulting drug levels and modulation of oxylipin pattern - Ostermann_2015_Prostaglandins.Other.Lipid.Mediat_121_131
Author(s) : Ostermann AI , Herbers J , Willenberg I , Chen R , Hwang SH , Greite R , Morisseau C , Gueler F , Hammock BD , Schebb NH
Ref : Prostaglandins Other Lipid Mediat , 121 :131 , 2015
Abstract : Epoxides from polyunsaturated fatty acids (PUFAs) are potent lipid mediators. In vivo stabilization of these epoxides by blockade of the soluble epoxide hydrolase (sEH) leads to anti-inflammatory, analgesic and normotensive effects. Therefore, sEH inhibitors (sEHi) are a promising new class of drugs. Herein, we characterized pharmacokinetic (PK) and pharmacodynamic properties of a commercially available potent sEHi 1-(1-propanoylpiperidin-4-yl)-3-[4-(trifluoromethoxy)phenyl]urea (TPPU). Cell culture studies suggest its high absorption and metabolic stability. Following administration in drinking water to rats (0.2, 1, and 5mg TPPU/L with 0.2% PEG400), TPPU's blood concentration increased dose dependently within the treatment period to reach an almost steady state after 8 days. TPPU was found in all the tissues tested. The linoleic epoxide/diol ratios in most tissues were dose dependently increased, indicating significant sEH inhibition. Overall, administration of TPPU with the drinking water led to systemic distribution as well as high drug levels and thus makes chronic sEH inhibition studies possible.
ESTHER : Ostermann_2015_Prostaglandins.Other.Lipid.Mediat_121_131
PubMedSearch : Ostermann_2015_Prostaglandins.Other.Lipid.Mediat_121_131
PubMedID: 26117215

Title : Inhibiting an Epoxide Hydrolase Virulence Factor from Pseudomonas aeruginosa Protects CFTR - Bahl_2015_Angew.Chem.Int.Ed.Engl_54_9881
Author(s) : Bahl CD , Hvorecny KL , Bomberger JM , Stanton BA , Hammock BD , Morisseau C , Madden DR
Ref : Angew Chem Int Ed Engl , 54 :9881 , 2015
Abstract : Opportunistic pathogens exploit diverse strategies to sabotage host defenses. Pseudomonas aeruginosa secretes the CFTR inhibitory factor Cif and thus triggers loss of CFTR, an ion channel required for airway mucociliary defense. However, the mechanism of action of Cif has remained unclear. It catalyzes epoxide hydrolysis, but there is no known role for natural epoxides in CFTR regulation. It was demonstrated that the hydrolase activity of Cif is strictly required for its effects on CFTR. A small-molecule inhibitor that protects this key component of the mucociliary defense system was also uncovered. These results provide a basis for targeting the distinctive virulence chemistry of Cif and suggest an unanticipated role of physiological epoxides in intracellular protein trafficking.
ESTHER : Bahl_2015_Angew.Chem.Int.Ed.Engl_54_9881
PubMedSearch : Bahl_2015_Angew.Chem.Int.Ed.Engl_54_9881
PubMedID: 26136396
Gene_locus related to this paper: pseae-PA2934

Title : Identification of potent inhibitors of the chicken soluble epoxide hydrolase - Shihadih_2015_Bioorg.Med.Chem.Lett_25_276
Author(s) : Shihadih DS , Harris TR , Yang J , Merzlikin O , Lee KS , Hammock BD , Morisseau C
Ref : Bioorganic & Medicinal Chemistry Lett , 25 :276 , 2015
Abstract : In vertebrates, soluble epoxide hydrolase (sEH) hydrolyzes natural epoxy-fatty acids (EpFAs), which are chemical mediators modulating inflammation, pain, and angiogenesis. Chick embryos are used to study angiogenesis, particularly its role in cardiovascular biology and pathology. To find potent and bio-stable inhibitors of the chicken sEH (chxEH) a library of human sEH inhibitors was screened. Derivatives of 1(adamantan-1-yl)-3-(trans-4-phenoxycyclohexyl) urea were found to be very potent tight binding inhibitors (KI <150pM) of chxEH while being relatively stable in chicken liver microsomes, suggesting their usefulness to study the role of EpFAs in chickens.
ESTHER : Shihadih_2015_Bioorg.Med.Chem.Lett_25_276
PubMedSearch : Shihadih_2015_Bioorg.Med.Chem.Lett_25_276
PubMedID: 25479771

Title : Omeprazole increases the efficacy of a soluble epoxide hydrolase inhibitor in a PGE2 induced pain model - Goswami_2015_Toxicol.Appl.Pharmacol_289_419
Author(s) : Goswami SK , Inceoglu B , Yang J , Wan D , Kodani SD , da Silva CA , Morisseau C , Hammock BD
Ref : Toxicol Appl Pharmacol , 289 :419 , 2015
Abstract : Epoxyeicosatrienoic acids (EETs) are potent endogenous analgesic metabolites produced from arachidonic acid by cytochrome P450s (P450s). Metabolism of EETs by soluble epoxide hydrolase (sEH) reduces their activity, while their stabilization by sEH inhibition decreases both inflammatory and neuropathic pain. Here, we tested the complementary hypothesis that increasing the level of EETs through induction of P450s by omeprazole (OME), can influence pain related signaling by itself, and potentiate the anti-hyperalgesic effect of sEH inhibitor. Rats were treated with OME (100mg/kg/day, p.o., 7days), sEH inhibitor TPPU (3mg/kg/day, p.o.) and OME (100mg/kg/day, p.o., 7days)+TPPU (3mg/kg/day, p.o., last 3days of OME dose) dissolved in vehicle PEG400, and their effect on hyperalgesia (increased sensitivity to pain) induced by PGE2 was monitored. While OME treatment by itself exhibited variable effects on PGE2 induced hyperalgesia, it strongly potentiated the effect of TPPU in the same assay. The significant decrease in pain with OME+TPPU treatment correlated with the increased levels of EETs in plasma and increased activities of P450 1A1 and P450 1A2 in liver microsomes. The results show that reducing catabolism of EETs with a sEH inhibitor yielded a stronger analgesic effect than increasing generation of EETs by OME, and combination of both yielded the strongest pain reducing effect under the condition of this study.
ESTHER : Goswami_2015_Toxicol.Appl.Pharmacol_289_419
PubMedSearch : Goswami_2015_Toxicol.Appl.Pharmacol_289_419
PubMedID: 26522832

Title : Soluble epoxide hydrolase inhibition improves coronary endothelial function and prevents the development of cardiac alterations in obese insulin-resistant mice - Roche_2015_Am.J.Physiol.Heart.Circ.Physiol_308_H1020
Author(s) : Roche C , Besnier M , Cassel R , Harouki N , Coquerel D , Guerrot D , Nicol L , Loizon E , Remy-Jouet I , Morisseau C , Mulder P , Ouvrard-Pascaud A , Madec AM , Richard V , Bellien J
Ref : American Journal of Physiology Heart Circ Physiol , 308 :H1020 , 2015
Abstract : This study addressed the hypothesis that inhibiting the soluble epoxide hydrolase (sEH)-mediated degradation of epoxy-fatty acids, notably epoxyeicosatrienoic acids, has an additional impact against cardiovascular damage in insulin resistance, beyond its previously demonstrated beneficial effect on glucose homeostasis. The cardiovascular and metabolic effects of the sEH inhibitor trans-4-[4-(3-adamantan-1-yl-ureido)-cyclohexyloxy]-benzoic acid (t-AUCB; 10 mg/l in drinking water) were compared with those of the sulfonylurea glibenclamide (80 mg/l), both administered for 8 wk in FVB mice subjected to a high-fat diet (HFD; 60% fat) for 16 wk. Mice on control chow diet (10% fat) and nontreated HFD mice served as controls. Glibenclamide and t-AUCB similarly prevented the increased fasting glycemia in HFD mice, but only t-AUCB improved glucose tolerance and decreased gluconeogenesis, without modifying weight gain. Moreover, t-AUCB reduced adipose tissue inflammation, plasma free fatty acids, and LDL cholesterol and prevented hepatic steatosis. Furthermore, only the sEH inhibitor improved endothelium-dependent relaxations to acetylcholine, assessed by myography in isolated coronary arteries. This improvement was related to a restoration of epoxyeicosatrienoic acid and nitric oxide pathways, as shown by the increased inhibitory effects of the nitric oxide synthase and cytochrome P-450 epoxygenase inhibitors l-NA and MSPPOH on these relaxations. Moreover, t-AUCB decreased cardiac hypertrophy, fibrosis, and inflammation and improved diastolic function, as demonstrated by the increased E/A ratio (echocardiography) and decreased slope of the end-diastolic pressure-volume relation (invasive hemodynamics). These results demonstrate that sEH inhibition improves coronary endothelial function and prevents cardiac remodeling and diastolic dysfunction in obese insulin-resistant mice.
ESTHER : Roche_2015_Am.J.Physiol.Heart.Circ.Physiol_308_H1020
PubMedSearch : Roche_2015_Am.J.Physiol.Heart.Circ.Physiol_308_H1020
PubMedID: 25724490

Title : Impact of soluble epoxide hydrolase inhibition on early kidney damage in hyperglycemic overweight mice - Roche_2015_Prostaglandins.Other.Lipid.Mediat_120_148
Author(s) : Roche C , Guerrot D , Harouki N , Duflot T , Besnier M , Remy-Jouet I , Renet S , Dumesnil A , Lejeune A , Morisseau C , Richard V , Bellien J
Ref : Prostaglandins Other Lipid Mediat , 120 :148 , 2015
Abstract : This study addressed the hypothesis that inhibition of the EETs degrading enzyme soluble epoxide hydrolase affords renal protection in the early stage of diabetic nephropathy. The renal effects of the sEH inhibitor t-AUCB (10mg/l in drinking water) were compared to those of the sulfonylurea glibenclamide (80mg/l), both administered for 8 weeks in FVB mice subjected to a high-fat diet (HFD, 60% fat) for 16 weeks. Mice on control chow diet (10% fat) and non-treated HFD mice served as controls. Compared with non-treated HFD mice, HFD mice treated with t-AUCB had a decreased EET degradation, as shown by their higher plasma EETs-to-DHETs ratio, and an increased EET production, as shown by the increase in EETs+DHETs levels, which was associated with induction of CYP450 epoxygenase expression. Both agents similarly reduced fasting glycemia but only t-AUCB prevented the increase in the urinary albumine-to-creatinine ratio in HFD mice. Histopathological analysis showed that t-AUCB reduced renal inflammation, which was associated with an increased mRNA expression of the NFkappaB inhibitor Ikappa identical with and related decrease in MCP-1, COX2 and VCAM-1 expressions. Finally, there was a marginally significant increase in reactive oxygen species production in HFD mice, together with an enhanced NOX2 expression. Both agents did not modify these parameters but t-AUCB increased the expression of the antioxidant enzyme superoxide dismutase 1. These results demonstrate that, independently from its glucose-lowering effect, sEH inhibition prevents microalbuminuria and renal inflammation in overweight hyperglycemic mice, suggesting that this pharmacological strategy could be useful in the management of diabetic nephropathy.
ESTHER : Roche_2015_Prostaglandins.Other.Lipid.Mediat_120_148
PubMedSearch : Roche_2015_Prostaglandins.Other.Lipid.Mediat_120_148
PubMedID: 26022136

Title : Peripheral FAAH and soluble epoxide hydrolase inhibitors are synergistically antinociceptive - Sasso_2015_Pharmacol.Res_97_7
Author(s) : Sasso O , Wagner K , Morisseau C , Inceoglu B , Hammock BD , Piomelli D
Ref : Pharmacol Res , 97 :7 , 2015
Abstract : We need better medicines to control acute and chronic pain. Fatty acid amide hydrolase (FAAH) and soluble epoxide hydrolase (sEH) catalyze the deactivating hydrolysis of two classes of bioactive lipid mediators--fatty acid ethanolamides (FAEs) and epoxidized fatty acids (EpFAs), respectively--which are biogenetically distinct but share the ability to attenuate pain responses and inflammation. In these experiments, we evaluated the antihyperalgesic activity of small-molecule inhibitors of FAAH and sEH, administered alone or in combination, in two pain models: carrageenan-induced hyperalgesia in mice and streptozocin-induced allodynia in rats. When administered separately, the sEH inhibitor 1-trifluoromethoxyphenyl-3-(1-propionylpiperidine-4-yl)urea (TPPU) and the peripherally restricted FAAH inhibitor URB937 were highly active in the two models. The combination TPPU plus URB937 was markedly synergistic, as assessed using isobolographic analyses. The results of these experiments reveal the existence of a possible functional crosstalk between FAEs and EpFAs in regulating pain responses. Additionally, the results suggest that combinations of sEH and FAAH inhibitors might be exploited therapeutically to achieve greater analgesic efficacy.
ESTHER : Sasso_2015_Pharmacol.Res_97_7
PubMedSearch : Sasso_2015_Pharmacol.Res_97_7
PubMedID: 25882247

Title : Epoxide hydrolase activities and epoxy fatty acids in the mosquito Culex quinquefasciatus - Xu_2015_Insect.Biochem.Mol.Biol_59_41
Author(s) : Xu J , Morisseau C , Yang J , Mamatha DM , Hammock BD
Ref : Insect Biochemistry & Molecular Biology , 59 :41 , 2015
Abstract : Culex mosquitoes have emerged as important model organisms for mosquito biology, and are disease vectors for multiple mosquito-borne pathogens, including West Nile virus. We characterized epoxide hydrolase activities in the mosquito Culex quinquefasciatus, which suggested multiple forms of epoxide hydrolases were present. We found EH activities on epoxy eicosatrienoic acids (EETs). EETs and other eicosanoids are well-established lipid signaling molecules in vertebrates. We showed EETs can be synthesized in vitro from arachidonic acids by mosquito lysate, and EETs were also detected in vivo both in larvae and adult mosquitoes by LC-MS/MS. The EH activities on EETs can be induced by blood feeding, and the highest activity was observed in the midgut of female mosquitoes. The enzyme activities on EETs can be inhibited by urea-based inhibitors designed for mammalian soluble epoxide hydrolases (sEH). The sEH inhibitors have been shown to play diverse biological roles in mammalian systems, and they can be useful tools to study the function of EETs in mosquitoes. Besides juvenile hormone metabolism and detoxification, insect epoxide hydrolases may also play a role in regulating lipid signaling molecules, such as EETs and other epoxy fatty acids, synthesized in vivo or obtained from blood feeding by female mosquitoes.
ESTHER : Xu_2015_Insect.Biochem.Mol.Biol_59_41
PubMedSearch : Xu_2015_Insect.Biochem.Mol.Biol_59_41
PubMedID: 25686802

Title : A photorhabdus natural product inhibits insect juvenile hormone epoxide hydrolase - Nollmann_2015_Chembiochem_16_766
Author(s) : Nollmann FI , Heinrich AK , Brachmann AO , Morisseau C , Mukherjee K , Casanova-Torres AM , Strobl F , Kleinhans D , Kinski S , Schultz K , Beeton ML , Kaiser M , Chu YY , Phan Ke L , Thanwisai A , Bozhuyuk KA , Chantratita N , Gotz F , Waterfield NR , Vilcinskas A , Stelzer EH , Goodrich-Blair H , Hammock BD , Bode HB
Ref : Chembiochem , 16 :766 , 2015
Abstract : Simple urea compounds ("phurealipids") have been identified from the entomopathogenic bacterium Photorhabdus luminescens, and their biosynthesis was elucidated. Very similar analogues of these compounds have been previously developed as inhibitors of juvenile hormone epoxide hydrolase (JHEH), a key enzyme in insect development and growth. Phurealipids also inhibit JHEH, and therefore phurealipids might contribute to bacterial virulence.
ESTHER : Nollmann_2015_Chembiochem_16_766
PubMedSearch : Nollmann_2015_Chembiochem_16_766
PubMedID: 25711603

Title : High salt diet modulates vascular response in A2AAR (+\/+) and A 2AAR (-\/-) mice: role of sEH, PPARgamma, and K ATP channels - Pradhan_2015_Mol.Cell.Biochem_404_87
Author(s) : Pradhan I , Ledent C , Mustafa SJ , Morisseau C , Nayeem MA
Ref : Molecular & Cellular Biochemistry , 404 :87 , 2015
Abstract : This study aims to investigate the signaling mechanism involved in HS-induced modulation of adenosine-mediated vascular tone in the presence or absence of adenosine A2A receptor (A2AAR). We hypothesized that HS-induced enhanced vascular relaxation through A2AAR and epoxyeicosatrienoic acid (EETs) is dependent on peroxisome proliferator-activated receptor gamma (PPARgamma) and ATP-sensitive potassium channels (KATP channels) in A2AAR(+/+) mice, while HS-induced vascular contraction to adenosine is dependent on soluble epoxide hydrolase (sEH) that degrades EETs in A2AAR(-/-) mice. Organ bath and Western blot techniques were conducted in HS (4 % NaCl) and normal salt (NS, 0.45 % NaCl)-fed A2AAR(+/+) and A2AAR(-/-) mouse aorta. We found that enhanced vasodilation to A2AAR agonist, CGS 21680, in HS-fed A2AAR(+/+) mice was blocked by PPARgamma antagonist (T0070907) and KATP channel blocker (Glibenclamide). Also, sEH inhibitor (AUDA)-dependent vascular relaxation was mitigated by PPARgamma antagonist. PPARgamma agonist (Rosiglitazone)-induced relaxation in HS-A2AAR(+/+) mice was attenuated by KATP channel blocker. Conversely, HS-induced contraction in A2AAR(-/-) mice was attenuated by sEH inhibitor. Overall, findings from this study that implicates the contribution of EETs, PPARgamma and KATP channels downstream of A2AAR to mediate enhanced vascular relaxation in response to HS diet while, role of sEH in mediating vascular contraction in HS-fed A2AAR(-/-) mice.
ESTHER : Pradhan_2015_Mol.Cell.Biochem_404_87
PubMedSearch : Pradhan_2015_Mol.Cell.Biochem_404_87
PubMedID: 25739357

Title : Potent natural soluble epoxide hydrolase inhibitors from Pentadiplandra brazzeana baillon: synthesis, quantification, and measurement of biological activities in vitro and in vivo - Kitamura_2015_PLoS.One_10_e0117438
Author(s) : Kitamura S , Morisseau C , Inceoglu B , Kamita SG , De Nicola GR , Nyegue M , Hammock BD
Ref : PLoS ONE , 10 :e0117438 , 2015
Abstract : We describe here three urea-based soluble epoxide hydrolase (sEH) inhibitors from the root of the plant Pentadiplandra brazzeana. The concentration of these ureas in the root was quantified by LC-MS/MS, showing that 1, 3-bis (4-methoxybenzyl) urea (MMU) is the most abundant (42.3 mug/g dry root weight). All of the ureas were chemically synthesized, and their inhibitory activity toward recombinant human and recombinant rat sEH was measured. The most potent compound, MMU, showed an IC50 of 92 nM via fluorescent assay and a Ki of 54 nM via radioactivity-based assay on human sEH. MMU effectively reduced inflammatory pain in a rat nociceptive pain assay. These compounds are among the most potent sEH inhibitors derived from natural sources. Moreover, inhibition of sEH by these compounds may mechanistically explain some of the therapeutic effects of P. brazzeana.
ESTHER : Kitamura_2015_PLoS.One_10_e0117438
PubMedSearch : Kitamura_2015_PLoS.One_10_e0117438
PubMedID: 25659109

Title : Protection from hypertension in mice by the Mediterranean diet is mediated by nitro fatty acid inhibition of soluble epoxide hydrolase - Charles_2014_Proc.Natl.Acad.Sci.U.S.A_111_8167
Author(s) : Charles RL , Rudyk O , Prysyazhna O , Kamynina A , Yang J , Morisseau C , Hammock BD , Freeman BA , Eaton P
Ref : Proc Natl Acad Sci U S A , 111 :8167 , 2014
Abstract : Soluble epoxide hydrolase (sEH) is inhibited by electrophilic lipids by their adduction to Cys521 proximal to its catalytic center. This inhibition prevents hydrolysis of the enzymes' epoxyeicosatrienoic acid (EET) substrates, so they accumulate inducing vasodilation to lower blood pressure (BP). We generated a Cys521Ser sEH redox-dead knockin (KI) mouse model that was resistant to this mode of inhibition. The electrophilic lipid 10-nitro-oleic acid (NO2-OA) inhibited hydrolase activity and also lowered BP in an angiotensin II-induced hypertension model in wild-type (WT) but not KI mice. Furthermore, EET/dihydroxy-epoxyeicosatrienoic acid isomer ratios were elevated in plasma from WT but not KI mice following NO2-OA treatment, consistent with the redox-dead mutant being resistant to inhibition by lipid electrophiles. sEH was inhibited in WT mice fed linoleic acid and nitrite, key constituents of the Mediterranean diet that elevates electrophilic nitro fatty acid levels, whereas KIs were unaffected. These observations reveal that lipid electrophiles such as NO2-OA mediate antihypertensive signaling actions by inhibiting sEH and suggest a mechanism accounting for protection from hypertension afforded by the Mediterranean diet.
ESTHER : Charles_2014_Proc.Natl.Acad.Sci.U.S.A_111_8167
PubMedSearch : Charles_2014_Proc.Natl.Acad.Sci.U.S.A_111_8167
PubMedID: 24843165

Title : Effects of soluble epoxide hydrolase deficiency on acute pancreatitis in mice - Bettaieb_2014_PLoS.One_9_e113019
Author(s) : Bettaieb A , Chahed S , Tabet G , Yang J , Morisseau C , Griffey S , Hammock BD , Haj FG
Ref : PLoS ONE , 9 :e113019 , 2014
Abstract : BACKGROUND: Acute pancreatitis (AP) is a frequent gastrointestinal disorder that causes significant morbidity, and its incidence has been progressively increasing. AP starts as a local inflammation in the pancreas that often leads to systemic inflammatory response and complications. Soluble epoxide hydrolase (sEH) is a cytosolic enzyme whose inhibition in murine models has beneficial effects in inflammatory diseases, but its significance in AP remains unexplored. METHODOLOGY/PRINCIPAL FINDINGS: To investigate whether sEH may have a causal role in AP we utilized Ephx2 knockout (KO) mice to determine the effects of sEH deficiency on cerulein- and arginine-induced AP. sEH expression increased at the protein and messenger RNA levels, as well as enzymatic activity in the early phase of cerulein- and arginine-induced AP in mice. In addition, amylase and lipase levels were lower in cerulein-treated Ephx2 KO mice compared with controls. Moreover, pancreatic mRNA and serum concentrations of the inflammatory cytokines IL-1B and IL-6 were lower in cerulein-treated Ephx2 KO mice compared with controls. Further, Ephx2 KO mice exhibited decreased cerulein- and arginine-induced NF-kappaB inflammatory response, MAPKs activation and decreased cell death. Conclusions -These findings demonstrate a novel role for sEH in the progression of cerulein- and arginine-induced AP.
ESTHER : Bettaieb_2014_PLoS.One_9_e113019
PubMedSearch : Bettaieb_2014_PLoS.One_9_e113019
PubMedID: 25402489

Title : Effect of soluble epoxide hydrolase polymorphism on substrate and inhibitor selectivity and dimer formation - Morisseau_2014_J.Lipid.Res_55_1131
Author(s) : Morisseau C , Wecksler AT , Deng C , Dong H , Yang J , Lee KS , Kodani SD , Hammock BD
Ref : J Lipid Res , 55 :1131 , 2014
Abstract : Epoxy FAs (EpFAs) are important lipid mediators that are mainly metabolized by soluble epoxide hydrolase (sEH). Thus, sEH inhibition is a promising therapeutic target to treat numerous ailments. Several sEH polymorphisms result in amino acid substitutions and alter enzyme activity. K55R and R287Q are associated with inflammatory, cardiovascular, and metabolic diseases. R287Q seems to affect sEH activity through reducing formation of a catalytically active dimer. Thus, understanding how these SNPs affect the selectivity of sEH for substrates and inhibitors is of potential clinical importance. We investigated the selectivity of four sEH SNPs toward a series of EpFAs and inhibitors. We found that the SNPs alter the catalytic activity of the enzyme but do not alter the relative substrate and inhibitor selectivity. We also determined their dimer/monomer constants (KD/M). The WT sEH formed a very tight dimer, with a KD/M in the low picomolar range. Only R287Q resulted in a large change of the KD/M. However, human tissue concentrations of sEH suggest that it is always in its dimer form independently of the SNP. These results suggest that the different biologies associated with K55R and R287Q are not explained by alteration in dimer formation or substrate selectivity.
ESTHER : Morisseau_2014_J.Lipid.Res_55_1131
PubMedSearch : Morisseau_2014_J.Lipid.Res_55_1131
PubMedID: 24771868

Title : Optimized inhibitors of soluble epoxide hydrolase improve in vitro target residence time and in vivo efficacy - Lee_2014_J.Med.Chem_57_7016
Author(s) : Lee KS , Liu JY , Wagner KM , Pakhomova S , Dong H , Morisseau C , Fu SH , Yang J , Wang P , Ulu A , Mate CA , Nguyen LV , Hwang SH , Edin ML , Mara AA , Wulff H , Newcomer ME , Zeldin DC , Hammock BD
Ref : Journal of Medicinal Chemistry , 57 :7016 , 2014
Abstract : Diabetes is affecting the life of millions of people. A large proportion of diabetic patients suffer from severe complications such as neuropathic pain, and current treatments for these complications have deleterious side effects. Thus, alternate therapeutic strategies are needed. Recently, the elevation of epoxy-fatty acids through inhibition of soluble epoxide hydrolase (sEH) was shown to reduce diabetic neuropathic pain in rodents. In this report, we describe a series of newly synthesized sEH inhibitors with at least 5-fold higher potency and doubled residence time inside both the human and rodent sEH enzyme than previously reported inhibitors. These inhibitors also have better physical properties and optimized pharmacokinetic profiles. The optimized inhibitor selected from this new series displayed improved efficacy of almost 10-fold in relieving pain perception in diabetic neuropathic rats as compared to the approved drug, gabapentin, and previously published sEH inhibitors. Therefore, these new sEH inhibitors could be an attractive alternative to treat diabetic neuropathy in humans.
ESTHER : Lee_2014_J.Med.Chem_57_7016
PubMedSearch : Lee_2014_J.Med.Chem_57_7016
PubMedID: 25079952
Gene_locus related to this paper: human-EPHX2

Title : Aberrant soluble epoxide hydrolase and oxylipin levels in a porcine arteriovenous graft stenosis model - Terry_2014_J.Vasc.Res_51_269
Author(s) : Terry CM , Carlson ML , He Y , Ulu A , Morisseau C , Blumenthal DK , Hammock BD , Cheung AK
Ref : J Vasc Res , 51 :269 , 2014
Abstract : Synthetic arteriovenous grafts (AVGs) used for hemodialysis frequently fail due to the development of neointimal hyperplasia (NH) at the vein-graft anastomosis. Inflammation and smooth-muscle cell (SMC) and myofibroblast proliferation and migration likely play an important role in the pathogenesis of NH. Epoxyeicosatrienoic acids (EETs), the products of the catabolism of arachidonic acid by cytochrome P450 enzymes, possess anti-inflammatory, antiproliferative, antimigratory and vasodilatory properties that should reduce NH. The degradation of vasculoprotective EETs is catalyzed by the enzyme, soluble epoxide hydrolase (sEH). sEH upregulation may thus contribute to NH development by the enhanced removal of vasculoprotective EETs. In this study, sEH, cytochrome P450 and EETs were examined after AVG placement in a porcine model to explore their potential roles in AVG stenosis. Increased sEH protein expression, decreased P450 epoxygenase activity and dysregulation of 5 oxylipin mediators were observed in the graft-venous anastomotic tissues when compared to control veins. Pharmacological inhibitors of sEH decreased the growth factor-induced migration of SMCs and fibroblasts, although they had no significant effect on the proliferation of these cells. These results provide insights on epoxide biology in vascular disorders and a rationale for the development of novel pharmacotherapeutic strategies to prevent AVG failure due to NH and stenosis.
ESTHER : Terry_2014_J.Vasc.Res_51_269
PubMedSearch : Terry_2014_J.Vasc.Res_51_269
PubMedID: 25196102

Title : Synthesis and functional survey of new Tacrine analogs modified with nitroxides or their precursors - Kalai_2014_Eur.J.Med.Chem_77C_343
Author(s) : Kalai T , Altman R , Maezawa I , Balog M , Morisseau C , Petrlova J , Hammock BD , Jin LW , Trudell JR , Voss JC , Hideg K
Ref : Eur Journal of Medicinal Chemistry , 77C :343 , 2014
Abstract : A series of new Tacrine analogs modified with nitroxides or pre-nitroxides on 9-amino group via methylene or piperazine spacers were synthesized; the nitroxide or its precursors were incorporated into the Tacrine scaffold. The new compounds were tested for their hydroxyl radical and peroxyl radical scavenging ability, acetylcholinesterase inhibitor activity and protection against Abeta-induced cytotoxicity. Based on these assays, we conclude that Tacrine analogs connected to five and six-membered nitroxides via piperazine spacers (9b, 9b/HCl and 12) exhibited the best activity, providing direction for further development of additional candidates with dual functionality (anti Alzheimer's and antioxidant).
ESTHER : Kalai_2014_Eur.J.Med.Chem_77C_343
PubMedSearch : Kalai_2014_Eur.J.Med.Chem_77C_343
PubMedID: 24657571

Title : Expression and characterization of an epoxide hydrolase from Anopheles gambiae with high activity on epoxy fatty acids - Xu_2014_Insect.Biochem.Mol.Biol_54C_42
Author(s) : Xu J , Morisseau C , Hammock BD
Ref : Insect Biochemistry & Molecular Biology , 54C :42 , 2014
Abstract : In insects, epoxide hydrolases (EHs) play critical roles in the metabolism of xenobiotic epoxides from the food resources and in the regulation of endogenous chemical mediators, such as juvenile hormones. Using the baculovirus expression system, we expressed and characterized an epoxide hydrolase from Anopheles gambiae (AgEH) that is distinct in evolutionary history from insect juvenile hormone epoxide hydrolases (JHEHs). We partially purified the enzyme by ion exchange chromatography and isoelectric focusing. The experimentally determined molecular weight and pI were estimated to be 35 kD and 6.3 respectively, different than the theoretical ones. The AgEH had the greatest activity on long chain epoxy fatty acids such as 14,15-epoxyeicosatrienoic acids (14,15-EET) and 9,10-epoxy-12Z-octadecenoic acids (9,10-EpOME or leukotoxin) among the substrates evaluated. Juvenile hormone III, a terpenoid insect growth regulator, was the next best substrate tested. The AgEH showed kinetics comparable to the mammalian soluble epoxide hydrolases, and the activity could be inhibited by AUDA [12-(3-adamantan-1-yl-ureido) dodecanoic acid], a urea-based inhibitor designed to inhibit the mammalian soluble epoxide hydrolases. The rabbit serum generated against the soluble epoxide hydrolase of Mus musculus can both cross-react with natural and denatured forms of the AgEH, suggesting immunologically they are similar. The study suggests there are mammalian sEH homologs in insects, and epoxy fatty acids may be important chemical mediators in insects.
ESTHER : Xu_2014_Insect.Biochem.Mol.Biol_54C_42
PubMedSearch : Xu_2014_Insect.Biochem.Mol.Biol_54C_42
PubMedID: 25173592

Title : Symmetric adamantyl-diureas as soluble epoxide hydrolase inhibitors - Burmistrov_2014_Bioorg.Med.Chem.Lett_24_2193
Author(s) : Burmistrov V , Morisseau C , Lee KS , Shihadih DS , Harris TR , Butov GM , Hammock BD
Ref : Bioorganic & Medicinal Chemistry Lett , 24 :2193 , 2014
Abstract : A series of inhibitors of the soluble epoxide hydrolase (sEH) containing two urea groups has been developed. Inhibition potency of the described compounds ranges from 2.0 muM to 0.4 nM. 1,6-(Hexamethylene)bis[(adamant-1-yl)urea] (3b) was found to be a potent slow tight binding inhibitor (IC50=0.5 nM) with a strong binding to sEH (Ki=3.1 nM) and a moderately long residence time on the enzyme (koff=1.05 x 10(-3) s(-1); t1/2=11 min).
ESTHER : Burmistrov_2014_Bioorg.Med.Chem.Lett_24_2193
PubMedSearch : Burmistrov_2014_Bioorg.Med.Chem.Lett_24_2193
PubMedID: 24685540

Title : Adenosine A2A receptor modulates vascular response in soluble epoxide hydrolase-null mice through CYP-epoxygenases and PPARgamma - Nayeem_2013_Am.J.Physiol.Regul.Integr.Comp.Physiol_304_R23
Author(s) : Nayeem MA , Pradhan I , Mustafa SJ , Morisseau C , Falck JR , Zeldin DC
Ref : American Journal of Physiology Regul Integr Comp Physiol , 304 :R23 , 2013
Abstract : The interaction between adenosine and soluble epoxide hydrolase (sEH) in vascular response is not known. Therefore, we hypothesized that lack of sEH in mice enhances adenosine-induced relaxation through A(2A) adenosine receptors (AR) via CYP-epoxygenases and peroxisome proliferator-activated receptor gamma (PPARgamma). sEH(-/-) showed an increase in A(2A) AR, CYP2J, and PPARgamma by 31%, 65%, and 36%, respectively, and a decrease in A(1)AR and PPARalpha (30% and 27%, respectively) vs. sEH(+/+). 5'-N-ethylcarboxamidoadenosine (NECA, an adenosine receptor agonist), CGS 21680 (A(2A) AR-agonist), and GW 7647 (PPARalpha-agonist)-induced responses were tested with nitro-l-arginine methyl ester (l-NAME) (NO-inhibitor; 10(-4) M), ZM-241385, SCH-58261 (A(2A) AR-antagonists; 10(-6) M), 14,15-epoxyeicosa-5(Z)-enoic acid (14,15-EEZE, an epoxyeicosatrienoic acid-antagonist; 10(-5) M), 12-(3-adamantan-1-yl-ureido) dodecanoic acid (AUDA; 10 muM) or trans-4-[4-(3-adamantan-1-yl-ureido)-cyclohexyloxy]-benzoic acid (t-AUCB, sEH-inhibitors; 10(-5) M), and T0070907 (PPARgamma-antagonist; 10(-7) M). In sEH(-/-) mice, ACh response was not different from sEH(+/+) (P > 0.05), and l-NAME blocked ACh-responses in both sEH(-/-) and sEH(+/+) mice (P < 0.05). NECA (10(-6) M)-induced relaxation was higher in sEH(-/-) (+12.94 +/- 3.2%) vs. sEH(+/+) mice (-5.35 +/- 5.2%); however, it was blocked by ZM-241385 (-22.42 +/- 1.9%) and SCH-58261(-30.04 +/- 4.2%). CGS-21680 (10(-6) M)-induced relaxation was higher in sEH(-/-) (+37.4 +/- 5.4%) vs. sEH(+/+) (+2.14 +/- 2.8%). l-NAME (sEH(-/-), +30.28 +/- 4.8%, P > 0.05) did not block CGS-21680-induced response, whereas 14,15-EEZE (-7.1 +/- 3.7%, P < 0.05) did. Also, AUDA and t-AUCB did not change CGS-21680-induced response in sEH(-/-) (P > 0.05), but reversed in sEH(+/+) (from +2.14 +/- 2.8% to +45.33 +/- 4.1%, and +63.37 +/- 7.2, respectively). PPARalpha-agonist did not relax as CGS 21680 (-2.48 +/- 1.1 vs. +37.4 +/- 5.4%) in sEH(-/-), and PPARgamma-antagonist blocked (from +37.4 +/- 5.4% to +9.40 +/- 3.1) CGS 21680-induced relaxation in sEH(-/-). Our data suggest that adenosine-induced relaxation in sEH(-/-) may depend on the upregulation of A(2A) AR, CYP2J, and PPARgamma, and the downregulation of A(1) AR and PPARalpha.
ESTHER : Nayeem_2013_Am.J.Physiol.Regul.Integr.Comp.Physiol_304_R23
PubMedSearch : Nayeem_2013_Am.J.Physiol.Regul.Integr.Comp.Physiol_304_R23
PubMedID: 23152114

Title : Characterization of HOVI-MEH1, a microsomal epoxide hydrolase from the glassy-winged sharpshooter Homalodisca vitripennis - Kamita_2013_Arch.Insect.Biochem.Physiol_83_171
Author(s) : Kamita SG , Oshita GH , Wang P , Morisseau C , Hammock BD , Nandety RS , Falk BW
Ref : Archives of Insect Biochemistry & Physiology , 83 :171 , 2013
Abstract : Epoxide hydrolase (EH) is an enzyme in the alpha/beta-hydrolase fold superfamily that uses a water molecule to transform an epoxide to its corresponding diol. In insects, EHs metabolize among other things critical developmental hormones called juvenile hormones (JHs). EHs also play roles in the detoxification of toxic compounds that are found in the insect's diet or environment. In this study, a full-length cDNA encoding an epoxide hydrolase, Hovi-mEH1, was obtained from the xylem-feeding insect Homalodisca vitripennis. H. vitripennis, commonly known as the glassy-winged sharpshooter, is an economically important vector of plant pathogenic bacteria such as Xylella fastidiosa. Hovi-mEH1 hydrolyzed the general EH substrates cis-stilbene oxide and trans-diphenylpropene oxide with specific activities of 47.5 +/- 6.2 and 1.3 +/- 0.5 nmol of diol formed min(-1) mg(-1) , respectively. Hovi-mEH1 metabolized JH III with a Vmax of 29.3 +/- 1.6 nmol min(-1) mg(-1) , kcat of 0.03 s(-1) , and KM of 13.8 +/- 2.0 muM. These Vmax and kcat values are similar to those of known JH metabolizing EHs from lepidopteran and coleopteran insects. Hovi-mEH1 showed 99.1% identity to one of three predicted EH-encoding sequences that were identified in the transcriptome of H. vitripennis. Of these three sequences only Hovi-mEH1 clustered with known JH metabolizing EHs. On the basis of biochemical, phylogenetic, and transcriptome analyses, we hypothesize that Hovi-mEH1 is a biologically relevantJH-metabolizing enzyme in H. vitripennis.
ESTHER : Kamita_2013_Arch.Insect.Biochem.Physiol_83_171
PubMedSearch : Kamita_2013_Arch.Insect.Biochem.Physiol_83_171
PubMedID: 23704009

Title : Substituted phenyl groups improve the pharmacokinetic profile and anti-inflammatory effect of urea-based soluble epoxide hydrolase inhibitors in murine models - Liu_2013_Eur.J.Pharm.Sci_48_619
Author(s) : Liu JY , Lin YP , Qiu H , Morisseau C , Rose TE , Hwang SH , Chiamvimonvat N , Hammock BD
Ref : Eur J Pharm Sci , 48 :619 , 2013
Abstract : Soluble epoxide hydrolase inhibitors (sEHIs) are anti-inflammatory, analgesic, anti-hypertensive, cardio- and renal-protective in multiple animal models. However, the earlier adamantyl-containing urea-based inhibitors are rapidly metabolized. Therefore, new potent inhibitors with the adamantyl group replaced by a substituted phenyl group were synthesized to presumptively offer better pharmacokinetic (PK) properties. Here we describe the improved PK profile of these inhibitors and the anti-inflammatory effect of the most promising one in a murine model. The PK profiles of inhibitors were determined following p.o. administration and serial bleeding in mice. The anti-inflammatory effect of 1-trifluoromethoxyphenyl-3-(1-propionylpiperidin-4-yl)urea (TPPU), the most promising inhibitor among the five sEHIs tested, was investigated in a lipopolysaccharide (LPS)-challenged murine model. The earlier broadly-used adamantyl-containing sEHI, trans-4-[4-(3-adamantan-1-yl-ureido)-cyclohexyloxy]-benzoic acid (t-AUCB), was used for comparison. Compared with the earlier adamantyl-containing urea-based inhibitors, substituted phenyl-containing urea-based inhibitors afford more favorable PK properties, such as higher Cmaxs, larger AUCs and longer t1/2s, which, as expected, show more stable metabolic stability. Moreover, oral administration of TPPU dramatically reversed the shifts caused by LPS-challenge in plasma levels of inflammatory cytokines, epoxides and corresponding diols, which is more potent than t-AUCB. The substituted phenyl-containing sEHIs are more metabolically stable than those with adamantyl group, resulting in more potent efficacy in vivo. This indicates a new strategy for development of sEHIs for further study toward clinical trials.
ESTHER : Liu_2013_Eur.J.Pharm.Sci_48_619
PubMedSearch : Liu_2013_Eur.J.Pharm.Sci_48_619
PubMedID: 23291046

Title : Development of On-line Liquid Chromatography-Biochemical Detection for Soluble Epoxide Hydrolase Inhibitors in Mixtures - Falck_2013_Chromatographia_76_13
Author(s) : Falck D , Schebb NH , Prihatiningtyas S , Zhang J , Heus F , Morisseau C , Kool J , Hammock BD , Niessen WM
Ref : Chromatographia , 76 :13 , 2013
Abstract : In this study, an end-point-based fluorescence assay for soluble epoxide hydrolase (sEH) was transformed into an on-line continuous-flow format. The on-line biochemical detection system (BCD) was coupled on-line to liquid chromatography (LC) to allow mixture analysis. The on-line BCD was based on a flow system wherein sEH activity was detected by competition of analytes with the substrate hydrolysis. The reaction product was measured by fluorescence detection. In parallel to the BCD data, UV and MS data were obtained through post-column splitting of the LC effluent. The buffer system and reagent concentrations were optimized resulting in a stable on-line BCD with a good assay window and good sensitivity (S/N > 60). The potency of known sEH inhibitors (sEHis) obtained by LC-BCD correlates well with published values. The LC-BCD system was applied to test how oxidative microsomal metabolism affects the potency of three sEHis. After incubation with pig liver microsomes, several metabolites of sEHis were characterized by MS, while their individual potencies were measured by BCD. For all compounds tested, active metabolites were observed. The developed method allows for the first time the detection of sEHis in mixtures providing new opportunities in the development of drug candidates.
ESTHER : Falck_2013_Chromatographia_76_13
PubMedSearch : Falck_2013_Chromatographia_76_13
PubMedID: 23526703

Title : Beneficial effects of inhibition of soluble epoxide hydrolase on glucose homeostasis and islet damage in a streptozotocin-induced diabetic mouse model - Chen_2013_Prostaglandins.Other.Lipid.Mediat_104-105_42
Author(s) : Chen L , Fan C , Zhang Y , Bakri M , Dong H , Morisseau C , Maddipati KR , Luo P , Wang CY , Hammock BD , Wang MH
Ref : Prostaglandins Other Lipid Mediat , 104-105 :42 , 2013
Abstract : Soluble epoxide hydrolase (sEH) is an enzyme involved in the metabolism of endogenous inflammatory and anti-apoptotic mediators. In the present study, we determined the effects of the inhibition of sEH on glucose homeostasis and islet damage in mice treated with streptozotocin (STZ), a model of chemical-induced diabetes. STZ increased daily water intake and decreased visceral (spleen and pancreas) weight in mice; sEH inhibition in STZ mice decreased water intake, but did not affect visceral weight. Hyperglycemia induced by STZ treatment in mice was attenuated by inhibiting sEH. The beneficial effects of sEH inhibition were accompanied, after 2 and 4 weeks of initial administration, by improving glucose tolerance. In contrast, sEH inhibition did not affect insulin tolerance. Using LC/MS analysis, neither STZ nor STZ plus sEH inhibition affected pancreatic and plasma ratios of epoxyeicosatrienoic acids (EETs) to dihydroxyeicosatrienoic acids (DHETs), an index of EETs levels. Western blot analysis showed that mouse cytochrome P450 (CYP) 2C enzymes are the major epoxygenases in islets. On day 5 after initial STZ treatment, STZ induced islet cell apoptosis, while sEH inhibition in STZ mice significantly reduced islet cell apoptosis. These studies provide pharmacological evidence that inhibiting sEH activity provides significant protection against islet beta-cell damage and improves glucose homeostasis in STZ-induced diabetes.
ESTHER : Chen_2013_Prostaglandins.Other.Lipid.Mediat_104-105_42
PubMedSearch : Chen_2013_Prostaglandins.Other.Lipid.Mediat_104-105_42
PubMedID: 23247129

Title : Role of epoxide hydrolases in lipid metabolism - Morisseau_2013_Biochimie_95_91
Author(s) : Morisseau C
Ref : Biochimie , 95 :91 , 2013
Abstract : Epoxide hydrolases (EH), enzymes present in all living organisms, transform epoxide-containing lipids to 1,2-diols by the addition of a molecule of water. Many of these oxygenated lipid substrates have potent biological activities: host defense, control of development, regulation of blood pressure, inflammation, and pain. In general, the bioactivity of these natural epoxides is significantly reduced upon metabolism to diols. Thus, through the regulation of the titer of lipid epoxides, EHs have important and diverse biological roles with profound effects on the physiological state of the host organism. This review will discuss the biological activity of key lipid epoxides in mammals. In addition, the use of EH specific inhibitors will be highlighted as possible therapeutic disease interventions.
ESTHER : Morisseau_2013_Biochimie_95_91
PubMedSearch : Morisseau_2013_Biochimie_95_91
PubMedID: 22722082

Title : Impact of soluble epoxide hydrolase and epoxyeicosanoids on human health - Morisseau_2013_Annu.Rev.Pharmacol.Toxicol_53_37
Author(s) : Morisseau C , Hammock BD
Ref : Annual Review of Pharmacology & Toxicology , 53 :37 , 2013
Abstract : The presence of epoxyeicosatrienoic acids (EETs) in tissues and their metabolism by soluble epoxide hydrolase (sEH) to 1,2-diols were first reported 30 years ago. However, appreciation of their importance in cell biology and physiology has greatly accelerated over the past decade with the discovery of metabolically stable inhibitors of sEH, the commercial availability of EETs, and the development of analytical methods for the quantification of EETs and their diols. Numerous roles of EETs in regulatory biology now are clear, and the value of sEH inhibition in various animal models of disease has been demonstrated. Here, we review these results and discuss how the pharmacological stabilization of EETs and other natural epoxy-fatty acids could lead to possible disease therapies.
ESTHER : Morisseau_2013_Annu.Rev.Pharmacol.Toxicol_53_37
PubMedSearch : Morisseau_2013_Annu.Rev.Pharmacol.Toxicol_53_37
PubMedID: 23020295

Title : Inhibition of soluble epoxide hydrolase by fulvestrant and sulfoxides - Morisseau_2013_Bioorg.Med.Chem.Lett_23_3818
Author(s) : Morisseau C , Pakhomova S , Hwang SH , Newcomer ME , Hammock BD
Ref : Bioorganic & Medicinal Chemistry Lett , 23 :3818 , 2013
Abstract : The soluble epoxide hydrolase (sEH) is a key enzyme in the metabolism of epoxy-fatty acids, signaling molecules involved in numerous biologies. Toward finding novel inhibitors of sEH, a library of known drugs was tested for inhibition of sEH. We found that fulvestrant, an anticancer agent, is a potent (KI=26 nM) competitive inhibitor of sEH. From this observation, we found that alkyl-sulfoxides represent a new kind of pharmacophore for the inhibition of sEH.
ESTHER : Morisseau_2013_Bioorg.Med.Chem.Lett_23_3818
PubMedSearch : Morisseau_2013_Bioorg.Med.Chem.Lett_23_3818
PubMedID: 23684894
Gene_locus related to this paper: human-EPHX2

Title : Frster resonance energy transfer competitive displacement assay for human soluble epoxide hydrolase - Lee_2013_Anal.Biochem_434_259
Author(s) : Lee KS , Morisseau C , Yang J , Wang P , Hwang SH , Hammock BD
Ref : Analytical Biochemistry , 434 :259 , 2013
Abstract : The soluble epoxide hydrolase (sEH), responsible for the hydrolysis of various fatty acid epoxides to their corresponding 1,2-diols, is becoming an attractive pharmaceutical target. These fatty acid epoxides, particularly epoxyeicosatrienoic acids (EETs), play an important role in human homeostatic and inflammation processes. Therefore, inhibition of human sEH, which stabilizes EETs in vivo, brings several beneficial effects to human health. Although there are several catalytic assays available to determine the potency of sEH inhibitors, measuring the in vitro inhibition constant (K(i)) for these inhibitors using catalytic assay is laborious. In addition, k(off), which has been recently suggested to correlate better with the in vivo potency of inhibitors, has never been measured for sEH inhibitors. To better measure the potency of sEH inhibitors, a reporting ligand, 1-(adamantan-1-yl)-3-(1-(2-(7-hydroxy-2-oxo-2H-chromen-4-yl)acetyl) piperidin-4-yl)urea (ACPU), was designed and synthesized. With ACPU, we have developed a Forster resonance energy transfer (FRET)-based competitive displacement assay using intrinsic tryptophan fluorescence from sEH. In addition, the resulting assay allows us to measure the K(i) values of very potent compounds to the picomolar level and to obtain relative k(off) values of the inhibitors. This assay provides additional data to evaluate the potency of sEH inhibitors.
ESTHER : Lee_2013_Anal.Biochem_434_259
PubMedSearch : Lee_2013_Anal.Biochem_434_259
PubMedID: 23219719

Title : Design, synthesis and anti-tuberculosis activity of 1-adamantyl-3-heteroaryl ureas with improved in vitro pharmacokinetic properties - North_2013_Bioorg.Med.Chem_21_2587
Author(s) : North EJ , Scherman MS , Bruhn DF , Scarborough JS , Maddox MM , Jones V , Grzegorzewicz A , Yang L , Hess T , Morisseau C , Jackson M , McNeil MR , Lee RE
Ref : Bioorganic & Medicinal Chemistry , 21 :2587 , 2013
Abstract : Out of the prominent global ailments, tuberculosis (TB) is still one of the leading causes of death worldwide due to infectious disease. Development of new drugs that shorten the current tuberculosis treatment time and have activity against drug resistant strains is of utmost importance. Towards these goals we have focused our efforts on developing novel anti-TB compounds with the general structure of 1-adamantyl-3-phenyl urea. This series is active against Mycobacteria and previous lead compounds were found to inhibit the membrane transporter MmpL3, the protein responsible for mycolic acid transport across the plasma membrane. However, these compounds suffered from poor in vitro pharmacokinetic (PK) profiles and they have a similar structure/SAR to inhibitors of human soluble epoxide hydrolase (sEH) enzymes. Therefore, in this study the further optimization of this compound class was driven by three factors: (1) to increase selectivity for anti-TB activity over human sEH activity, (2) to optimize PK profiles including solubility and (3) to maintain target inhibition. A new series of 1-adamantyl-3-heteroaryl ureas was designed and synthesized replacing the phenyl substituent of the original series with pyridines, pyrimidines, triazines, oxazoles, isoxazoles, oxadiazoles and pyrazoles. This study produced lead isoxazole, oxadiazole and pyrazole substituted adamantyl ureas with improved in vitro PK profiles, increased selectivity and good anti-TB potencies with sub mug/mL minimum inhibitory concentrations.
ESTHER : North_2013_Bioorg.Med.Chem_21_2587
PubMedSearch : North_2013_Bioorg.Med.Chem_21_2587
PubMedID: 23498915

Title : Cloning and characterization of a microsomal epoxide hydrolase from Heliothis virescens - Kamita_2013_Insect.Biochem.Mol.Biol_43_219
Author(s) : Kamita SG , Yamamoto K , Dadala MM , Pha K , Morisseau C , Escaich A , Hammock BD
Ref : Insect Biochemistry & Molecular Biology , 43 :219 , 2013
Abstract : Epoxide hydrolases (EHs) are alpha/beta-hydrolase fold superfamily enzymes that convert epoxides to 1,2-trans diols. In insects EHs play critical roles in the metabolism of toxic compounds and allelochemicals found in the diet and for the regulation of endogenous juvenile hormones (JHs). In this study we obtained a full-length cDNA, hvmeh1, from the generalist feeder Heliothis virescens that encoded a highly active EH, Hv-mEH1. Of the 10 different EH substrates that were tested, Hv-mEH1 showed the highest specific activity (1180 nmol min(-1) mg(-1)) for a 1,2-disubstituted epoxide-containing fluorescent substrate. This specific activity was more than 25- and 3900-fold higher than that for the general EH substrates cis-stilbene oxide and trans-stilbene oxide, respectively. Although phylogenetic analysis placed Hv-mEH1 in a clade with some lepidopteran JH metabolizing EHs (JHEHs), JH III was a relatively poor substrate for Hv-mEH1. Hv-mEH1 showed a unique substrate selectivity profile for the substrates tested in comparison to those of MsJHEH, a well-characterized JHEH from Manduca sexta, and hmEH, a human microsomal EH. Hv-mEH1 also showed unique enzyme inhibition profiles to JH-like urea, JH-like secondary amide, JH-like primary amide, and non-JH-like primary amide compounds in comparison to MsJHEH and hmEH. Although Hv-mEH1 is capable of metabolizing JH III, our findings suggest that this enzymatic activity does not play a significant role in the metabolism of JH in the caterpillar. The ability of Hv-mEH1 to rapidly hydrolyze 1,2-disubstituted epoxides suggests that it may play roles in the metabolism of fatty acid epoxides such as those that are commonly found in the diet of Heliothis.
ESTHER : Kamita_2013_Insect.Biochem.Mol.Biol_43_219
PubMedSearch : Kamita_2013_Insect.Biochem.Mol.Biol_43_219
PubMedID: 23276675
Gene_locus related to this paper: helvi-l7r9x8

Title : Soluble Epoxide Hydrolase Deficiency or Inhibition Attenuates Diet-induced Endoplasmic Reticulum Stress in Liver and Adipose Tissue - Bettaieb_2013_J.Biol.Chem_288_14189
Author(s) : Bettaieb A , Nagata N , Aboubechara D , Chahed S , Morisseau C , Hammock BD , Haj FG
Ref : Journal of Biological Chemistry , 288 :14189 , 2013
Abstract : Soluble epoxide hydrolase (sEH) is a cytosolic enzyme whose inhibition has beneficial effects in cardiovascular, inflammatory, and metabolic diseases in murine models. Mice with targeted deletion or pharmacological inhibition of sEH exhibit improved insulin signaling in liver and adipose tissue. Herein, we assessed the role of sEH in regulating endoplasmic reticulum (ER) stress in liver and adipose tissue. We report that sEH expression was increased in the livers and adipose tissue of mice fed a high fat diet, the adipose tissue of overweight humans, and palmitate-treated cells. Importantly, sEH deficiency or inhibition in mice attenuated chronic high fat diet-induced ER stress in liver and adipose tissue. Similarly, pharmacological inhibition of sEH in HepG2 cells and 3T3-L1 adipocytes mitigated chemical-induced ER stress and activation of JNK, p38, and cell death. In addition, insulin signaling was enhanced in HepG2 cells treated with sEH substrates and attenuated in cells treated with sEH products. In summary, these findings demonstrate that sEH is a physiological modulator of ER stress and a potential target for mitigating complications associated with obesity.
ESTHER : Bettaieb_2013_J.Biol.Chem_288_14189
PubMedSearch : Bettaieb_2013_J.Biol.Chem_288_14189
PubMedID: 23576437

Title : A potent soluble epoxide hydrolase inhibitor, t-AUCB, acts through PPARgamma to modulate the function of endothelial progenitor cells from patients with acute myocardial infarction - Xu_2013_Int.J.Cardiol_167_1298
Author(s) : Xu DY , Davis BB , Wang ZH , Zhao SP , Wasti B , Liu ZL , Li N , Morisseau C , Chiamvimonvat N , Hammock BD
Ref : Int J Cardiol , 167 :1298 , 2013
Abstract : BACKGROUND: Epoxyeicosatrienoic acids (EETs) are natural angiogenic mediators regulated by soluble epoxide hydrolase (sEH). Inhibitors of sEH can stabilize EETs levels and were reported to reduce atherosclerosis and inhibit myocardial infarction in animal models. In this work, we investigated whether increasing EETs with the sEH inhibitor t-AUCB would increase angiogenesis related function in endothelial progenitor cells (EPCs) from patients with acute myocardial infarction (AMI). METHODS AND
RESULTS: EPCs were isolated from 50 AMI patients and 50 healthy subjects (control). EPCs were treated with different concentrations of t-AUCB for 24h with or without peroxisome proliferator activated receptor gamma (PPARgamma) inhibitor GW9662. Migration of EPCs was assayed in trans-well chambers. Angiogenesis assays were performed using a Matrigel-Matrix in vitro model. The expression of vascular endothelial growth factor (VEGF), hypoxia-inducible factor 1alpha (HIF-1alpha) mRNA and protein in EPCs was measured by real-time PCR or Western blot, respectively. Also, the concentration of EETs in the culture supernatant was detected by ELISA. The activity of EPCs in the AMI patient group was reduced compared to healthy controls. Whereas increasing EET levels with t-AUCB promoted a dose dependent angiogenesis and migration in EPCs from AMI patients. Additionally, the t-AUCB dose dependently increased the expression of the angiogenic factors VEGF and HIF-alpha. Lastly, we provide evidence that these effects were PPARgamma dependent. CONCLUSION: The results demonstrate that the sEH inhibitor positively modulated the functions of EPCs in patients with AMI through the EETs-PPARgamma pathway. The present study suggests the potential utility of sEHi in the therapy of ischemic heart disease.
ESTHER : Xu_2013_Int.J.Cardiol_167_1298
PubMedSearch : Xu_2013_Int.J.Cardiol_167_1298
PubMedID: 22525341

Title : Synthesis and biological evaluation of sorafenib- and regorafenib-like sEH inhibitors - Hwang_2013_Bioorg.Med.Chem.Lett_23_3732
Author(s) : Hwang SH , Wecksler AT , Zhang G , Morisseau C , Nguyen LV , Fu SH , Hammock BD
Ref : Bioorganic & Medicinal Chemistry Lett , 23 :3732 , 2013
Abstract : To reduce the pro-angiogenic effects of sEH inhibition, a structure-activity relationship (SAR) study was performed by incorporating structural features of the anti-angiogenic multi-kinase inhibitor sorafenib into soluble epoxide hydrolase (sEH) inhibitors. The structural modifications of this series of molecules enabled the altering of selectivity towards the pro-angiogenic kinases C-RAF and vascular endothelial growth factor receptor-2 (VEGFR-2), while retaining their sEH inhibition. As a result, sEH inhibitors with greater potency against C-RAF and VEGFR-2 were obtained. Compound 4 (t-CUPM) possesses inhibition potency higher than sorafenib towards sEH but similar against C-RAF and VEGFR-2. Compound 7 (t-CUCB) selectively inhibits sEH, while inhibiting HUVEC cell proliferation, a potential anti-angiogenic property, without liver cancer cell cytotoxicity. The data presented suggest a potential rational approach to control the angiogenic responses stemming from sEH inhibition.
ESTHER : Hwang_2013_Bioorg.Med.Chem.Lett_23_3732
PubMedSearch : Hwang_2013_Bioorg.Med.Chem.Lett_23_3732
PubMedID: 23726028

Title : Synthesis and structure-activity relationship of piperidine-derived non-urea soluble epoxide hydrolase inhibitors - Pecic_2013_Bioorg.Med.Chem.Lett_23_417
Author(s) : Pecic S , Pakhomova S , Newcomer ME , Morisseau C , Hammock BD , Zhu Z , Rinderspacher A , Deng SX
Ref : Bioorganic & Medicinal Chemistry Lett , 23 :417 , 2013
Abstract : A series of potent amide non-urea inhibitors of soluble epoxide hydrolase (sEH) is disclosed. The inhibition of soluble epoxide hydrolase leads to elevated levels of epoxyeicosatrienoic acids (EETs), and thus inhibitors of sEH represent one of a novel approach to the development of vasodilatory and anti-inflammatory drugs. Structure-activities studies guided optimization of a lead compound, identified through high-throughput screening, gave rise to sub-nanomolar inhibitors of human sEH with stability in human liver microsomal assay suitable for preclinical development.
ESTHER : Pecic_2013_Bioorg.Med.Chem.Lett_23_417
PubMedSearch : Pecic_2013_Bioorg.Med.Chem.Lett_23_417
PubMedID: 23237835
Gene_locus related to this paper: human-EPHX2

Title : Pharmacological inhibition of soluble epoxide hydrolase ameliorates diet-induced metabolic syndrome in rats - Iyer_2012_Exp.Diabetes.Res_2012_758614
Author(s) : Iyer A , Kauter K , Alam MA , Hwang SH , Morisseau C , Hammock BD , Brown L
Ref : Exp Diabetes Res , 2012 :758614 , 2012
Abstract : The signs of metabolic syndrome following chronic excessive macronutrient intake include body weight gain, excess visceral adipose deposition, hyperglycaemia, glucose and insulin intolerances, hypertension, dyslipidaemia, endothelial damage, cardiovascular hypertrophy, inflammation, ventricular contractile dysfunction, fibrosis, and fatty liver disease. Recent studies show increased activity of soluble epoxide hydrolase (sEH) during obesity and metabolic dysfunction. We have tested whether sEH inhibition has therapeutic potential in a rat model of diet-induced metabolic syndrome. In these high-carbohydrate, high-fat-fed rats, chronic oral treatment with trans-4-[4-(3-adamantan-1-ylureido)-cyclohexyloxy]-benzoic acid (t-AUCB), a potent sEH inhibitor, alleviated the signs of metabolic syndrome in vivo including glucose, insulin, and lipid abnormalities, changes in pancreatic structure, increased systolic blood pressure, cardiovascular structural and functional abnormalities, and structural and functional changes in the liver. The present study describes the pharmacological responses to this selective sEH inhibitor in rats with the signs of diet-induced metabolic syndrome.
ESTHER : Iyer_2012_Exp.Diabetes.Res_2012_758614
PubMedSearch : Iyer_2012_Exp.Diabetes.Res_2012_758614
PubMedID: 22007192

Title : Design, synthesis and evaluation of non-urea inhibitors of soluble epoxide hydrolase - Pecic_2012_Bioorg.Med.Chem.Lett_22_601
Author(s) : Pecic S , Deng SX , Morisseau C , Hammock BD , Landry DW
Ref : Bioorganic & Medicinal Chemistry Lett , 22 :601 , 2012
Abstract : Inhibition of soluble epoxide hydrolase (sEH) has been proposed as a new pharmaceutical approach for treating hypertension and vascular inflammation. The most potent sEH inhibitors reported in literature to date are urea derivatives. However, these compounds have limited pharmacokinetic profiles. We investigated non-urea amide derivatives as sEH inhibitors and identified a potent human sEH inhibitor 14-34 having potency comparable to urea-based inhibitors.
ESTHER : Pecic_2012_Bioorg.Med.Chem.Lett_22_601
PubMedSearch : Pecic_2012_Bioorg.Med.Chem.Lett_22_601
PubMedID: 22079754

Title : Screening a library of 1600 adamantyl ureas for anti-Mycobacterium tuberculosis activity in vitro and for better physical chemical properties for bioavailability - Scherman_2012_Bioorg.Med.Chem_20_3255
Author(s) : Scherman MS , North EJ , Jones V , Hess TN , Grzegorzewicz AE , Kasagami T , Kim IH , Merzlikin O , Lenaerts AJ , Lee RE , Jackson M , Morisseau C , McNeil MR
Ref : Bioorganic & Medicinal Chemistry , 20 :3255 , 2012
Abstract : Adamantyl ureas were previously identified as a group of compounds active against Mycobacterium tuberculosis in culture with minimum inhibitor concentrations (MICs) below 0.1 mug/ml. These compounds have been shown to target MmpL3, a protein involved in secretion of trehalose mono-mycolate. They also inhibit both human soluble epoxide hydrolase (hsEH) and M. tuberculosis epoxide hydrolases. However, active compounds to date have high cLogP's and are poorly soluble, leading to low bioavailability and thus limiting any therapeutic application. In this study, a library of 1600 ureas (mostly adamantyl ureas), which were synthesized for the purpose of increasing the bioavailability of inhibitors of hsEH, was screened for activity against M. tuberculosis. 1-Adamantyl-3-phenyl ureas with a polar para substituent were found to retain moderate activity against M. tuberculosis and one of these compounds was shown to be present in serum after oral administration to mice. However, neither it, nor a closely related analog, reduced M. tuberculosis infection in mice. No correlation between in vitro potency against M. tuberculosis and the hsEH inhibition were found supporting the concept that activity against hsEH and M. tuberculosis can be separated. Also there was a lack of correlation with cLogP and inhibition of the growth of M. tuberculosis. Finally, members of two classes of adamantyl ureas that contained polar components to increase their bioavailability, but lacked efficacy against growing M. tuberculosis, were found to taken up by the bacterium as effectively as a highly active apolar urea suggesting that these modifications to increase bioavailability affected the interaction of the urea against its target rather than making them unable to enter the bacterium.
ESTHER : Scherman_2012_Bioorg.Med.Chem_20_3255
PubMedSearch : Scherman_2012_Bioorg.Med.Chem_20_3255
PubMedID: 22522007

Title : Inhibition of mycolic acid transport across the Mycobacterium tuberculosis plasma membrane - Grzegorzewicz_2012_Nat.Chem.Biol_8_334
Author(s) : Grzegorzewicz AE , Pham H , Gundi VA , Scherman MS , North EJ , Hess T , Jones V , Gruppo V , Born SE , Kordulakova J , Chavadi SS , Morisseau C , Lenaerts AJ , Lee RE , McNeil MR , Jackson M
Ref : Nat Chemical Biology , 8 :334 , 2012
Abstract : New chemotherapeutics active against multidrug-resistant Mycobacterium tuberculosis are urgently needed. We report on the identification of an adamantyl urea compound that shows potent bactericidal activity against M. tuberculosis and a unique mode of action, namely the abolition of the translocation of mycolic acids from the cytoplasm, where they are synthesized to the periplasmic side of the plasma membrane and are in turn transferred onto cell wall arabinogalactan or used in the formation of virulence-associated, outer membrane, trehalose-containing glycolipids. Whole-genome sequencing of spontaneous-resistant mutants of M. tuberculosis selected in vitro followed by genetic validation experiments revealed that our prototype inhibitor targets the inner membrane transporter MmpL3. Conditional gene expression of mmpL3 in mycobacteria and analysis of inhibitor-treated cells validate MmpL3 as essential for mycobacterial growth and support the involvement of this transporter in the translocation of trehalose monomycolate across the plasma membrane.
ESTHER : Grzegorzewicz_2012_Nat.Chem.Biol_8_334
PubMedSearch : Grzegorzewicz_2012_Nat.Chem.Biol_8_334
PubMedID: 22344175

Title : EH3 (ABHD9): the first member of a new epoxide hydrolase family with high activity for fatty acid epoxides - Decker_2012_J.Lipid.Res_53_2038
Author(s) : Decker M , Adamska M , Cronin A , Di Giallonardo F , Burgener J , Marowsky A , Falck JR , Morisseau C , Hammock BD , Gruzdev A , Zeldin DC , Arand M
Ref : J Lipid Res , 53 :2038 , 2012
Abstract : Epoxide hydrolases are a small superfamily of enzymes important for the detoxification of chemically reactive xenobiotic epoxides and for the processing of endogenous epoxides that act as signaling molecules. Here, we report the identification of two human epoxide hydrolases: EH3 and EH4. They share 45% sequence identity, thus representing a new family of mammalian epoxide hydrolases. Quantitative RT-PCR from mouse tissue indicates strongest EH3 expression in lung, skin, and upper gastrointestinal tract. The recombinant enzyme shows a high turnover number with 8,9-, 11,12-, and 14,15-epoxyeicosatrienoic acid (EET), as well as 9,10-epoxyoctadec-11-enoic acid (leukotoxin). It is inhibited by a subclass of N,N'-disubstituted urea derivatives, including 12-(3-adamantan-1-yl-ureido)-dodecanoic acid, 1-cyclohexyl-3-dodecylurea, and 1-(1-acetylpiperidin-4-yl)-3-(4-(trifluoromethoxy)phenyl)urea, compounds so far believed to be selective inhibitors of mammalian soluble epoxide hydrolase (sEH). Its sensitivity to this subset of sEH inhibitors may have implications on the pharmacologic profile of these compounds. This is particularly relevant because sEH is a potential drug target, and clinical trials are under way exploring the value of sEH inhibitors in the treatment of hypertension and diabetes type II.
ESTHER : Decker_2012_J.Lipid.Res_53_2038
PubMedSearch : Decker_2012_J.Lipid.Res_53_2038
PubMedID: 22798687
Gene_locus related to this paper: human-EPHX3 , human-EPHX4

Title : Development of monoclonal antibodies to human microsomal epoxide hydrolase and analysis of preneoplastic antigen-like molecules - Duan_2012_Toxicol.Appl.Pharmacol_260_17
Author(s) : Duan H , Yoshimura K , Kobayashi N , Sugiyama K , Sawada J , Saito Y , Morisseau C , Hammock BD , Akatsuka T
Ref : Toxicol Appl Pharmacol , 260 :17 , 2012
Abstract : Microsomal epoxide hydrolase (mEH) is a drug metabolizing enzyme which resides on the endoplasmic reticulum (ER) membrane and catalyzes the hydration of reactive epoxide intermediates that are formed by cytochrome P450s. mEH is also thought to have a role in bile acid transport on the plasma membrane of hepatocytes. It is speculated that efficient execution of such multiple functions is secured by its orientation and association with cytochrome P450 enzymes on the ER membrane and formation of a multiple transport system on the plasma membrane. In certain disease status, mEH loses its association with the membrane and can be detected as distinct antigens in the cytosol of preneoplastic foci of liver (preneoplastic antigen), in the serum in association with hepatitis C virus infection (AN antigen), or in some brain tumors. To analyze the antigenic structures of mEH in physiological and pathological conditions, we developed monoclonal antibodies against different portions of mEH. Five different kinds of antibodies were obtained: three, anti-N-terminal portions; one anti-C-terminal; and one, anti-conformational epitope. By combining these antibodies, we developed antigen detection methods which are specific to either the membrane-bound form or the linearized form of mEH. These methods detected mEH in the culture medium released from a hepatocellular carcinoma cell line and a glioblastoma cell line, which was found to be a multimolecular complex with a unique antigenic structure different from that of the membrane-bound form of mEH. These antibodies and antigen detection methods may be useful to study pathological changes of mEH in various human diseases.
ESTHER : Duan_2012_Toxicol.Appl.Pharmacol_260_17
PubMedSearch : Duan_2012_Toxicol.Appl.Pharmacol_260_17
PubMedID: 22310175

Title : Development of fluorescent substrates for microsomal epoxide hydrolase and application to inhibition studies - Morisseau_2011_Anal.Biochem_414_154
Author(s) : Morisseau C , Bernay M , Escaich A , Sanborn JR , Lango J , Hammock BD
Ref : Analytical Biochemistry , 414 :154 , 2011
Abstract : The microsomal epoxide hydrolase (mEH) plays a significant role in the metabolism of numerous xenobiotics. In addition, it has a potential role in sexual development and bile acid transport, and it is associated with a number of diseases such as emphysema, spontaneous abortion, eclampsia, and several forms of cancer. Toward developing chemical tools to study the biological role of mEH, we designed and synthesized a series of absorbent and fluorescent substrates. The highest activity for both rat and human mEH was obtained with the fluorescent substrate cyano(6-methoxy-naphthalen-2-yl)methyl glycidyl carbonate (11). An in vitro inhibition assay using this substrate ranked a series of known inhibitors similarly to the assay that used radioactive cis-stilbene oxide but with a greater discrimination between inhibitors. These results demonstrate that the new fluorescence-based assay is a useful tool for the discovery of structure-activity relationships among mEH inhibitors. Furthermore, this substrate could also be used for the screening chemical library with high accuracy and with a Z' value of approximately 0.7. This new assay permits a significant decrease in labor and cost and also offers the advantage of a continuous readout. However, it should not be used with crude enzyme preparations due to interfering reactions.
ESTHER : Morisseau_2011_Anal.Biochem_414_154
PubMedSearch : Morisseau_2011_Anal.Biochem_414_154
PubMedID: 21371418
Gene_locus related to this paper: human-EPHX1 , ratno-hyep

Title : Improvement of endothelium-dependent vasodilations by SKA-31 and SKA-20, activators of small- and intermediate-conductance Ca2+ -activated K+ -channels - Hasenau_2011_Acta.Physiol.(Oxf)_203_117
Author(s) : Hasenau AL , Nielsen G , Morisseau C , Hammock BD , Wulff H , Kohler R
Ref : Acta Physiol (Oxf) , 203 :117 , 2011
Abstract : AIM: Endothelial membrane hyperpolarization mediated by KCa3.1 and KCa2.3 channels has been demonstrated to initiate endothelium-derived hyperpolarizing factor (EDHF)-type vasodilations. Moreover, pharmacological potentiation of KCa3.1/KCa2.3 channels has been suggested to improve EDHF-type vasodilations. Herein, we determined whether the KCa3.1/KCa2.3 activator SKA-31 and its derivative SKA-20 improve endothelial dysfunction in KCa3.1-/- and NOS3-/- mice.
METHODS: Membrane potentials were measured using patch-clamp electrophysiology on carotid artery (CA) endothelial cells (CAEC) from wild-type (wt) and KCa3.1-/- mice. Endothelium-dependent vasodilations were determined by pressure myography in CA.
RESULTS: SKA-31 (1 mum) activated KCa3.1 and KCa2.3 channels and induced membrane hyperpolarization in CAEC of wt (DeltaMP -45 mV). These responses were significantly reduced in CAEC of KCa3.1-/- (DeltaMP -8 mV). SKA-31 (200 nm, 500 nm) and SKA-20 (300 nm) significantly enhanced EDHF vasodilations in wt. SKA-20 also improved vasodilations during NO synthesis. In KCa3.1-/-, the defective EDHF vasodilations were unchanged at 200 nm SKA-31, but were significantly improved at 500 nm. EDHF vasodilations were slightly enhanced at 300 nm SKA-20, but vasodilations during NO synthesis were unchanged. SKA-31 (500 nm) enhanced the impaired endothelium-dependent vasodilation in NOS3-/- mice twofold. Pharmacological inhibition of the soluble epoxide hydrolase by t-AUCB (1 mum) in contrast did not increase ACh-induced EDHF- or NO-mediated vasodilations in wt and KCa3.1-/-. CONCLUSION: Normal and defective endothelium-dependent vasodilations in murine carotid arteries can be improved by pharmacological enhancement of KCa3.1/KCa2.3 functions. These findings further support the concept that pharmacological activation of endothelial KCa2.3/KCa3.1 could offer a novel endothelium-specific antihypertensive strategy.
ESTHER : Hasenau_2011_Acta.Physiol.(Oxf)_203_117
PubMedSearch : Hasenau_2011_Acta.Physiol.(Oxf)_203_117
PubMedID: 21362152

Title : Redox regulation of soluble epoxide hydrolase by 15-deoxy-delta-prostaglandin J2 controls coronary hypoxic vasodilation - Charles_2011_Circ.Res_108_324
Author(s) : Charles RL , Burgoyne JR , Mayr M , Weldon SM , Hubner N , Dong H , Morisseau C , Hammock BD , Landar A , Eaton P
Ref : Circulation Research , 108 :324 , 2011
Abstract : RATIONALE: 15-Deoxy-Delta-prostaglandin (15d-PG)J(2) is an electrophilic oxidant that dilates the coronary vasculature. This lipid can adduct to redox active protein thiols to induce oxidative posttranslational modifications that modulate protein and tissue function. OBJECTIVE: To investigate the role of oxidative protein modifications in 15d-PGJ(2)-mediated coronary vasodilation and define the distal signaling pathways leading to enhanced perfusion. METHODS AND RESULTS: Proteomic screening with biotinylated 15d-PGJ(2) identified novel vascular targets to which it adducts, most notably soluble epoxide hydrolase (sEH). 15d-PGJ(2) inhibited sEH by specifically adducting to a highly conserved thiol (Cys521) adjacent to the catalytic center of the hydrolase. Indeed a Cys521Ser sEH "redox-dead" mutant was resistant to 15d-PGJ(2)-induced hydrolase inhibition. 15d-PGJ(2) dilated coronary vessels and a role for hydrolase inhibition was supported by 2 structurally different sEH antagonists each independently inducing vasorelaxation. Furthermore, 15d-PGJ(2) and sEH antagonists also increased coronary effluent epoxyeicosatrienoic acids consistent with their vasodilatory actions. Indeed 14,15-EET alone induced relaxation and 15d-PGJ(2)-mediated vasodilation was blocked by the EET receptor antagonist 14,15-epoxyeicosa-5(Z)-enoic acid (14,15-EEZE). Additionally, the coronary vasculature of sEH-null mice was basally dilated compared to wild-type controls and failed to vasodilate in response to 15d-PGJ(2). Coronary vasodilation to hypoxia in wild-types was accompanied by 15d-PGJ(2) adduction to and inhibition of sEH. Consistent with the importance of hydrolase inhibition, sEH-null mice failed to vasodilate during hypoxia. CONCLUSION: This represents a new paradigm for the regulation of sEH by an endogenous lipid, which is integral to the fundamental physiological response of coronary hypoxic vasodilation.
ESTHER : Charles_2011_Circ.Res_108_324
PubMedSearch : Charles_2011_Circ.Res_108_324
PubMedID: 21164107

Title : Synthesis and structure-activity relationship studies of urea-containing pyrazoles as dual inhibitors of cyclooxygenase-2 and soluble epoxide hydrolase - Hwang_2011_J.Med.Chem_54_3037
Author(s) : Hwang SH , Wagner KM , Morisseau C , Liu JY , Dong H , Wecksler AT , Hammock BD
Ref : Journal of Medicinal Chemistry , 54 :3037 , 2011
Abstract : A series of dual inhibitors containing a 1,5-diarylpyrazole and a urea were designed, synthesized, and evaluated as novel COX-2/sEH dual inhibitors in vitro using recombinant enzyme assays and in vivo using a lipopolysaccharide (LPS) induced model of pain in rats. The best inhibition potencies and selectivity for sEH and COX-2 over COX-1 were obtained with compounds (21b, 21i, and 21j) in which both the 1,5-diaryl-pyrazole group and the urea group are linked with a three-methylene group. Compound 21i showed the best pharmacokinetic profiles in both mice and rats (higher AUC and longer half-life). Following subcutaneous administration at 10 mg/kg, compound 21i exhibited antiallodynic activity that is more effective than the same dose of either a COX-2 inhibitor (celecoxib) or a sEH inhibitor (t-AUCB) alone, as well as coadministration of both inhibitors. Thus, these novel dual inhibitors exhibited enhanced in vivo antiallodynic activity in a nociceptive behavioral assay.
ESTHER : Hwang_2011_J.Med.Chem_54_3037
PubMedSearch : Hwang_2011_J.Med.Chem_54_3037
PubMedID: 21434686

Title : Development of an online SPE-LC-MS-based assay using endogenous substrate for investigation of soluble epoxide hydrolase (sEH) inhibitors - Schebb_2011_Anal.Bioanal.Chem_400_1359
Author(s) : Schebb NH , Huby M , Morisseau C , Hwang SH , Hammock BD
Ref : Anal Bioanal Chem , 400 :1359 , 2011
Abstract : Soluble epoxide hydrolase (sEH) is a promising therapeutic target for the treatment of hypertension, pain, and inflammation-related diseases. In order to enable the development of sEH inhibitors (sEHIs), assays are needed for determination of their potency. Therefore, we developed a new method utilizing an epoxide of arachidonic acid (14(15)-EpETrE) as substrate. Incubation samples were directly injected without purification into an online solid phase extraction (SPE) liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS-MS) setup allowing a total run time of only 108 s for a full gradient separation. Analytes were extracted from the matrix within 30 s by turbulent flow chromatography. Subsequently, a full gradient separation was carried out on a 50X2.1 mm RP-18 column filled with 1.7 mum core-shell particles. The analytes were detected with high sensitivity by ESI-MS-MS in SRM mode. The substrate 14(15)-EpETrE eluted at a stable retention time of 96 +/- 1 s and its sEH hydrolysis product 14,15-DiHETrE at 63 +/- 1 s with narrow peak width (full width at half maximum height: 1.5 +/- 0.1 s). The analytical performance of the method was excellent, with a limit of detection of 2 fmol on column, a linear range of over three orders of magnitude, and a negligible carry-over of 0.1% for 14,15-DiHETrE. The enzyme assay was carried out in a 96-well plate format, and near perfect sigmoidal dose-response curves were obtained for 12 concentrations of each inhibitor in only 22 min, enabling precise determination of IC(50) values. In contrast with other approaches, this method enables quantitative evaluation of potent sEHIs with picomolar potencies because only 33 pmol L(-1) sEH were used in the reaction vessel. This was demonstrated by ranking ten compounds by their activity; in the fluorescence method all yielded IC(50) <= 1 nmol L(-1). Comparison of 13 inhibitors with IC(50) values >1 nmol L(-1) showed a good correlation with the fluorescence method (linear correlation coefficient 0.9, slope 0.95, Spearman's rho 0.9). For individual compounds, however, up to eightfold differences in potencies between this and the fluorescence method were obtained. Therefore, enzyme assays using natural substrate, as described here, are indispensable for reliable determination of structure-activity relationships for sEH inhibition.
ESTHER : Schebb_2011_Anal.Bioanal.Chem_400_1359
PubMedSearch : Schebb_2011_Anal.Bioanal.Chem_400_1359
PubMedID: 21479549

Title : The structure-activity relationship of urea derivatives as anti-tuberculosis agents - Brown_2011_Bioorg.Med.Chem_19_5585
Author(s) : Brown JR , North EJ , Hurdle JG , Morisseau C , Scarborough JS , Sun D , Kordulakova J , Scherman MS , Jones V , Grzegorzewicz A , Crew RM , Jackson M , McNeil MR , Lee RE
Ref : Bioorganic & Medicinal Chemistry , 19 :5585 , 2011
Abstract : The treatment of tuberculosis is becoming more difficult due to the ever increasing prevalence of drug resistance. Thus, it is imperative that novel anti-tuberculosis agents, with unique mechanisms of action, be discovered and developed. The direct anti-tubercular testing of a small compound library led to discovery of adamantyl urea hit compound 1. In this study, the hit was followed up through the synthesis of an optimization library. This library was generated by systematically replacing each section of the molecule with a similar moiety until a clear structure-activity relationship was obtained with respect to anti-tubercular activity. The best compounds in this series contained a 1-adamantyl-3-phenyl urea core and had potent activity against Mycobacterium tuberculosis plus an acceptable therapeutic index. It was noted that the compounds identified and the pharmacophore developed is consistent with inhibitors of epoxide hydrolase family of enzymes. Consequently, the compounds were tested for inhibition of representative epoxide hydrolases: M. tuberculosis EphB and EphE; and human soluble epoxide hydrolase. Many of the optimized inhibitors showed both potent EphB and EphE inhibition suggesting the antitubercular activity is through inhibition of multiple epoxide hydrolase enzymes. The inhibitors also showed potent inhibition of humans soluble epoxide hydrolase, but limited cytotoxicity suggesting that future studies must be towards increasing the selectivity of epoxide hydrolase inhibition towards the M. tuberculosis enzymes.
ESTHER : Brown_2011_Bioorg.Med.Chem_19_5585
PubMedSearch : Brown_2011_Bioorg.Med.Chem_19_5585
PubMedID: 21840723

Title : Analgesia mediated by soluble epoxide hydrolase inhibitors is dependent on cAMP - Inceoglu_2011_Proc.Natl.Acad.Sci.U.S.A_108_5093
Author(s) : Inceoglu B , Wagner K , Schebb NH , Morisseau C , Jinks SL , Ulu A , Hegedus C , Rose T , Brosnan R , Hammock BD
Ref : Proc Natl Acad Sci U S A , 108 :5093 , 2011
Abstract : Pain is a major health concern even though numerous analgesic agents are available. Side effects and lack of wide-spectrum efficacy of current drugs justify efforts to better understand pain mechanisms. Stabilization of natural epoxy-fatty acids (EFAs) through inhibition of the soluble epoxide hydrolase (sEH) reduces pain. However, in the absence of an underlying painful state, inhibition of sEH is ineffective. Surprisingly, a pain-mediating second messenger, cAMP, interacts with natural EFAs and regulates the analgesic activity of sEH inhibitors. Concurrent inhibition of sEH and phosphodiesterase (PDE) dramatically reduced acute pain in rodents. Our findings demonstrate a mechanism of action of cAMP and EFAs in the pathophysiology of pain. Furthermore, we demonstrate that inhibition of various PDE isozymes, including PDE4, lead to significant increases in EFA levels through a mechanism independent of sEH, suggesting that the efficacy of commercial PDE inhibitors could result in part from increasing EFAs. The cross-talk between the two major pathways-one mediated by cAMP and the other by EFAs-paves the way to new approaches to understand and control pain.
ESTHER : Inceoglu_2011_Proc.Natl.Acad.Sci.U.S.A_108_5093
PubMedSearch : Inceoglu_2011_Proc.Natl.Acad.Sci.U.S.A_108_5093
PubMedID: 21383170

Title : Inhibition or deletion of soluble epoxide hydrolase prevents hyperglycemia, promotes insulin secretion, and reduces islet apoptosis - Luo_2010_J.Pharmacol.Exp.Ther_334_430
Author(s) : Luo P , Chang HH , Zhou Y , Zhang S , Hwang SH , Morisseau C , Wang CY , Inscho EW , Hammock BD , Wang MH
Ref : Journal of Pharmacology & Experimental Therapeutics , 334 :430 , 2010
Abstract : Soluble epoxide hydrolase (sEH) is an enzyme involved in the metabolism of endogenous inflammatory and antiapoptotic mediators. However, the roles of sEH in diabetes and the pancreas are unknown. Our aims were to determine whether sEH is involved in the regulation of hyperglycemia in diabetic mice and to investigate the reasons for the regulation of insulin secretion by sEH deletion or inhibition in islets. We used two separate approaches, targeted disruption of Ephx2 gene [sEH knockout (KO)] and a selective inhibitor of sEH [trans-4-[4-(3-adamantan-1-ylureido)-cyclohexyloxy]-benzoic acid (t-AUCB)], to assess the role of sEH in glucose and insulin homeostasis in streptozotocin (STZ) mice. We also examined the effects of sEH KO or t-AUCB on glucose-stimulated insulin secretion (GSIS) and intracellular calcium levels in islets. Hyperglycemia in STZ mice was prevented by both sEH KO and t-AUCB. In addition, STZ mice with sEH KO had improved glucose tolerance. More important, when insulin levels were assessed by hyperglycemic clamp study, sEH KO was found to promote insulin secretion. In addition, sEH KO and t-AUCB treatment augmented islet GSIS. Islets with sEH KO had a greater intracellular calcium influx when challenged with high glucose or KCl in the presence of diazoxide. Moreover, sEH KO reduced islet cell apoptosis in STZ mice. These results show not only that sEH KO and its inhibition prevent hyperglycemia in diabetes, but also that sEH KO enhances islet GSIS through the amplifying pathway and decreases islet cell apoptosis in diabetes.
ESTHER : Luo_2010_J.Pharmacol.Exp.Ther_334_430
PubMedSearch : Luo_2010_J.Pharmacol.Exp.Ther_334_430
PubMedID: 20439437

Title : Incorporation of piperazino functionality into 1,3-disubstituted urea as the tertiary pharmacophore affording potent inhibitors of soluble epoxide hydrolase with improved pharmacokinetic properties - Huang_2010_J.Med.Chem_53_8376
Author(s) : Huang SX , Li HY , Liu JY , Morisseau C , Hammock BD , Long YQ
Ref : Journal of Medicinal Chemistry , 53 :8376 , 2010
Abstract : The inhibition of the mammalian soluble epoxide hydrolase (sEH) is a promising new therapy in the treatment of hypertension, inflammation, and other disorders. However, the problems of limited water solubility, high melting point, and low metabolic stability complicated the development of 1,3-disubstituted urea-based sEH inhibitors. The current study explored the introduction of the substituted piperazino group as the tertiary pharmacophore, which resulted in substantial improvements in pharmacokinetic parameters over previously reported 1-adamantylurea based inhibitors while retaining high potency. The SAR studies revealed that the meta- or para-substituted phenyl spacer and N(4)-acetyl or sulfonyl substituted piperazine were optimal structures for achieving high potency and good physical properties. The 1-(4-(4-(4-acetylpiperazin-1-yl)butoxy)phenyl)-3-adamantan-1-yl urea (29c) demonstrated excellent in vivo pharmacokinetic properties in mice: T1/2 =14 h, Cmax = 84 nM, AUC = 40 200 nM.min, and IC50 = 7.0 nM against human sEH enzyme.
ESTHER : Huang_2010_J.Med.Chem_53_8376
PubMedSearch : Huang_2010_J.Med.Chem_53_8376
PubMedID: 21070033

Title : Soluble epoxide hydrolase deficiency attenuates neointima formation in the femoral cuff model of hyperlipidemic mice - Revermann_2010_Arterioscler.Thromb.Vasc.Biol_30_909
Author(s) : Revermann M , Schloss M , Barbosa-Sicard E , Mieth A , Liebner S , Morisseau C , Geisslinger G , Schermuly RT , Fleming I , Hammock BD , Brandes RP
Ref : Arterioscler Thromb Vasc Biol , 30 :909 , 2010
Abstract : OBJECTIVE: Epoxyeicosatrienoic acids (EETs) have antiinflammatory effects and are required for normal endothelial function. The soluble epoxide hydrolase (sEH) metabolizes EETs to their less active diols. We hypothesized that knockout and inhibition of sEH prevents neointima formation in hyperlipidemic ApoE(-/-) mice. METHODS AND RESULTS: Inhibition of sEH by 12-(3-adamantan-1-yl-ureido) dodecanoic acid or knockout of the enzyme significantly increased plasma EET levels. sEH activity was detectable in femoral and carotid arteries. sEH knockout or inhibition resulted in a significant reduction of neointima formation in the femoral artery cuff model but not following carotid artery ligation. Although macrophage infiltration occurred abundantly at the site of cuff placement in both sEH(+/+) and sEH(-/-), the expression of proinflammatory genes was significantly reduced in femoral arteries from sEH(-/-) mice. Moreover, an in vivo 5-bromo-2'-deoxyuridine assay revealed that smooth muscle cell proliferation at the site of cuff placement was attenuated in sEH knockout and sEH inhibitor-treated animals. CONCLUSION: These observations suggest that inhibition of sEH prevents vascular remodeling in an inflammatory model but not in a blood flow-dependent model of neointima formation.
ESTHER : Revermann_2010_Arterioscler.Thromb.Vasc.Biol_30_909
PubMedSearch : Revermann_2010_Arterioscler.Thromb.Vasc.Biol_30_909
PubMedID: 20224052

Title : Naturally occurring monoepoxides of eicosapentaenoic acid and docosahexaenoic acid are bioactive antihyperalgesic lipids - Morisseau_2010_J.Lipid.Res_51_3481
Author(s) : Morisseau C , Inceoglu B , Schmelzer K , Tsai HJ , Jinks SL , Hegedus CM , Hammock BD
Ref : J Lipid Res , 51 :3481 , 2010
Abstract : Beneficial physiological effects of long-chain n-3 polyunsaturated fatty acids are widely accepted but the mechanism(s) by which these fatty acids act remains unclear. Herein, we report the presence, distribution, and regulation of the levels of n-3 epoxy-fatty acids by soluble epoxide hydrolase (sEH) and a direct antinociceptive role of n-3 epoxy-fatty acids, specifically those originating from docosahexaenoic acid (DHA). The monoepoxides of the C18:1 to C22:6 fatty acids in both the n-6 and n-3 series were prepared and the individual regioisomers purified. The kinetic constants of the hydrolysis of the pure regioisomers by sEH were measured. Surprisingly, the best substrates are the mid-chain DHA epoxides. We also demonstrate that the DHA epoxides are present in considerable amounts in the rat central nervous system. Furthermore, using an animal model of pain associated with inflammation, we show that DHA epoxides, but neither the parent fatty acid nor the corresponding diols, selectively modulate nociceptive pathophysiology. Our findings support an important function of epoxy-fatty acids in the n-3 series in modulating nociceptive signaling. Consequently, the DHA and eicosapentaenoic acid epoxides may be responsible for some of the beneficial effects associated with dietary n-3 fatty acid intake.
ESTHER : Morisseau_2010_J.Lipid.Res_51_3481
PubMedSearch : Morisseau_2010_J.Lipid.Res_51_3481
PubMedID: 20664072

Title : Crystal structure of the CFTR inhibitory factor Cif reveals novel active-site features of an epoxide hydrolase virulence factor. - Bahl_2010_J.Bacteriol_192_1785
Author(s) : Bahl CD , Morisseau C , Bomberger JM , Stanton BA , Hammock BD , O'Toole GA , Madden DR
Ref : Journal of Bacteriology , 192 :1785 , 2010
Abstract : Cystic fibrosis transmembrane conductance regulator (CFTR) inhibitory factor (Cif) is a virulence factor secreted by Pseudomonas aeruginosa that reduces the quantity of CFTR in the apical membrane of human airway epithelial cells. Initial sequence analysis suggested that Cif is an epoxide hydrolase (EH), but its sequence violates two strictly conserved EH motifs and is also compatible with other alpha/beta hydrolase family members with diverse substrate specificities. To investigate the mechanistic basis of Cif activity, we have determined its structure at 1.8 A resolution by X-ray crystallography. The catalytic triad consists of residues Asp129, His297, and Glu153, which are conserved across the family of EHs. At other positions, sequence deviations from canonical EH active-site motifs are stereochemically conservative. Furthermore, detailed enzymatic analysis confirms that Cif catalyzes the hydrolysis of epoxide compounds, with specific activity against both epibromohydrin and cis-stilbene oxide, but with a relatively narrow range of substrate selectivity. Although closely related to two other classes of alpha/beta hydrolase in both sequence and structure, Cif does not exhibit activity as either a haloacetate dehalogenase or a haloalkane dehalogenase. Reassessment of the structural and functional consequences of the H269A mutation suggests that Cif's effect on host-cell CFTR expression may require hydrolysis of an extended endogenous epoxide substrate.
ESTHER : Bahl_2010_J.Bacteriol_192_1785
PubMedSearch : Bahl_2010_J.Bacteriol_192_1785
PubMedID: 20118260
Gene_locus related to this paper: pseae-PA2934

Title : Pharmacokinetic screening of soluble epoxide hydrolase inhibitors in dogs - Tsai_2010_Eur.J.Pharm.Sci_40_222
Author(s) : Tsai HJ , Hwang SH , Morisseau C , Yang J , Jones PD , Kasagami T , Kim IH , Hammock BD
Ref : Eur J Pharm Sci , 40 :222 , 2010
Abstract : Epoxyeicosatrienoic acids that have anti-hypertensive and anti-inflammatory properties are mainly metabolized by soluble epoxide hydrolase (sEH, EC 3.3.2.3). Therefore, sEH has emerged as a therapeutic target for treating various cardiovascular diseases and inflammatory pain. N,N'-Disubstituted ureas are potent sEH inhibitors in vitro. However, in vivo usage of early sEH inhibitors has been limited by their low bioavailability and poor physiochemical properties. Therefore, a group of highly potent compounds with more drug-like physiochemical properties were evaluated by monitoring their plasma profiles in dogs treated orally with sEH inhibitors. Urea compounds with an adamantyl or a 4-trifluoromethoxyphenyl group on one side and a piperidyl or a cyclohexyl ether group on the other side of the urea function showed pharmacokinetic profiles with high plasma concentrations and long half lives. In particular, the inhibitor trans-4-[4-(3-adamantan-1-yl-ureido)-cyclohexyloxy]-benzoic acid (t-AUCB) not only is very potent with good physiochemical properties, but also shows high oral bioavailability for doses ranging from 0.01 to 1mg/kg. This compound is also very potent against the sEH of several mammals, suggesting that t-AUCB will be an excellent tool to evaluate the biology of sEH in multiple animal models. Such compounds may also be a valuable lead for the development of veterinary therapeutics.
ESTHER : Tsai_2010_Eur.J.Pharm.Sci_40_222
PubMedSearch : Tsai_2010_Eur.J.Pharm.Sci_40_222
PubMedID: 20359531

Title : 1-Aryl-3-(1-acylpiperidin-4-yl)urea inhibitors of human and murine soluble epoxide hydrolase: structure-activity relationships, pharmacokinetics, and reduction of inflammatory pain - Rose_2010_J.Med.Chem_53_7067
Author(s) : Rose TE , Morisseau C , Liu JY , Inceoglu B , Jones PD , Sanborn JR , Hammock BD
Ref : Journal of Medicinal Chemistry , 53 :7067 , 2010
Abstract : 1,3-Disubstituted ureas possessing a piperidyl moiety have been synthesized to investigate their structure-activity relationships as inhibitors of the human and murine soluble epoxide hydrolase (sEH). Oral administration of 13 1-aryl-3-(1-acylpiperidin-4-yl)urea inhibitors in mice revealed substantial improvements in pharmacokinetic parameters over previously reported 1-adamantylurea based inhibitors. For example, 1-(1-(cyclopropanecarbonyl)piperidin-4-yl)-3-(4-(trifluoromethoxy)phenyl)urea (52) showed a 7-fold increase in potency, a 65-fold increase in C(max), and a 3300-fold increase in AUC over its adamantane analogue 1-(1-adamantyl)-3-(1-propionylpiperidin-4-yl)urea (2). This novel sEH inhibitor showed a 1000-fold increase in potency when compared to morphine by reducing hyperalgesia as measured by mechanical withdrawal threshold using the in vivo carrageenan induced inflammatory pain model.
ESTHER : Rose_2010_J.Med.Chem_53_7067
PubMedSearch : Rose_2010_J.Med.Chem_53_7067
PubMedID: 20812725

Title : Inhibition of the soluble epoxide hydrolase promotes albuminuria in mice with progressive renal disease - Jung_2010_PLoS.One_5_e11979
Author(s) : Jung O , Jansen F , Mieth A , Barbosa-Sicard E , Pliquett RU , Babelova A , Morisseau C , Hwang SH , Tsai C , Hammock BD , Schaefer L , Geisslinger G , Amann K , Brandes RP
Ref : PLoS ONE , 5 :e11979 , 2010
Abstract : Epoxyeicotrienoic acids (EETs) are cytochrome P450-dependent anti-hypertensive and anti-inflammatory derivatives of arachidonic acid, which are highly abundant in the kidney and considered reno-protective. EETs are degraded by the enzyme soluble epoxide hydrolase (sEH) and sEH inhibitors are considered treatment for chronic renal failure (CRF). We determined whether sEH inhibition attenuates the progression of CRF in the 5/6-nephrectomy model (5/6-Nx) in mice. 5/6-Nx mice were treated with a placebo, an ACE-inhibitor (Ramipril, 40 mg/kg), the sEH-inhibitor cAUCB or the CYP-inhibitor fenbendazole for 8 weeks. 5/6-Nx induced hypertension, albuminuria, glomerulosclerosis and tubulo-interstitial damage and these effects were attenuated by Ramipril. In contrast, cAUCB failed to lower the blood pressure and albuminuria was more severe as compared to placebo. Plasma EET-levels were doubled in 5/6 Nx-mice as compared to sham mice receiving placebo. Renal sEH expression was attenuated in 5/6-Nx mice but cAUCB in these animals still further increased the EET-level. cAUCB also increased 5-HETE and 15-HETE, which derive from peroxidation or lipoxygenases. Similar to cAUCB, CYP450 inhibition increased HETEs and promoted albuminuria. Thus, sEH-inhibition failed to elicit protective effects in the 5/6-Nx model and showed a tendency to aggravate the disease. These effects might be consequence of a shift of arachidonic acid metabolism into the lipoxygenase pathway.
ESTHER : Jung_2010_PLoS.One_5_e11979
PubMedSearch : Jung_2010_PLoS.One_5_e11979
PubMedID: 20694143

Title : Function of phenylalanine 259 and threonine 314 within the substrate binding pocket of the juvenile hormone esterase of Manduca sexta - Kamita_2010_Biochemistry_49_3733
Author(s) : Kamita SG , Wogulis MD , Law CS , Morisseau C , Tanaka H , Huang H , Wilson DK , Hammock BD
Ref : Biochemistry , 49 :3733 , 2010
Abstract : Juvenile hormone (JH) is a key insect developmental hormone that is found at low nanomolar levels in larval insects. The methyl ester of JH is hydrolyzed in many insects by an esterase that shows high specificity for JH. We have previously determined a crystal structure of the JH esterase (JHE) of the tobacco hornworm Manduca sexta (MsJHE) [Wogulis, M., Wheelock, C. E., Kamita, S. G., Hinton, A. C., Whetstone, P. A., Hammock, B. D., and Wilson, D. K. (2006) Biochemistry 45, 4045-4057]. Our molecular modeling indicates that JH fits very tightly within the substrate binding pocket of MsJHE. This tight fit places two noncatalytic amino acid residues, Phe-259 and Thr-314, within the appropriate distance and geometry to potentially interact with the alpha,beta-unsaturated ester and epoxide, respectively, of JH. These residues are highly conserved in numerous biologically active JHEs. Kinetic analyses of mutants of Phe-259 or Thr-314 indicate that these residues contribute to the low K(M) that MsJHE shows for JH. This low K(M), however, comes at the cost of reduced substrate turnover. Neither nucleophilic attack of the resonance-stabilized ester by the catalytic serine nor the availability of a water molecule for attack of the acyl-enzyme intermediate appears to be a rate-determining step in the hydrolysis of JH by MsJHE. We hypothesize that the release of the JH acid metabolite from the substrate binding pocket limits the catalytic cycle. Our findings also demonstrate that chemical bond strength does not necessarily correlate with how reactive the bond will be to metabolism.
ESTHER : Kamita_2010_Biochemistry_49_3733
PubMedSearch : Kamita_2010_Biochemistry_49_3733
PubMedID: 20307057

Title : Attenuation of cisplatin nephrotoxicity by inhibition of soluble epoxide hydrolase - Parrish_2009_Cell.Biol.Toxicol_25_217
Author(s) : Parrish AR , Chen G , Burghardt RC , Watanabe T , Morisseau C , Hammock BD
Ref : Cell Biol Toxicol , 25 :217 , 2009
Abstract : Cisplatin is a highly effective chemotherapeutic agent against many tumors; however, it is also a potent nephrotoxicant. Given that there have been no significant advances in our ability to clinically manage acute renal failure since the advent of dialysis, the development of novel strategies to ablate nephrotoxicity would represent a significant development. In this study, we investigated the ability of an inhibitor of soluble epoxide hydrolase (sEH), n-butyl ester of 12-(3-adamantan-1-yl-ureiido)-dodecanoic acid (nbAUDA), to attenuate cisplatin-induced nephrotoxicity. nbAUDA is quickly converted to AUDA and results in maintenance of high AUDA levels in vivo. Subcutaneous administration of 40 mg/kg of nbAUDA to C3H mice every 24 h resulted in elevated blood levels of AUDA; this protocol was also associated with attenuation of nephrotoxicity induced by cisplatin (intraperitoneal injection) as assessed by BUN levels and histological evaluation of kidneys. This is the first report of the use of sEH inhibitors to protect against acute nephrotoxicity and suggests a therapeutic potential of these compounds.
ESTHER : Parrish_2009_Cell.Biol.Toxicol_25_217
PubMedSearch : Parrish_2009_Cell.Biol.Toxicol_25_217
PubMedID: 18386137

Title : Pharmacokinetic optimization of four soluble epoxide hydrolase inhibitors for use in a murine model of inflammation - Liu_2009_Br.J.Pharmacol_156_284
Author(s) : Liu JY , Tsai HJ , Hwang SH , Jones PD , Morisseau C , Hammock BD
Ref : British Journal of Pharmacology , 156 :284 , 2009
Abstract : BACKGROUND AND PURPOSE: Early soluble epoxide hydrolase inhibitors (sEHIs) such as 12-(3-adamantan-1-yl-ureido)-dodecanoic acid (AUDA) are effective anti-hypertensive and anti-inflammatory agents in various animal models. However, their poor metabolic stability and limited water solubility make them difficult to use pharmacologically. Here we present the evaluation of four sEHIs for improved pharmacokinetic properties and the anti-inflammatory effects of one sEHI. EXPERIMENTAL APPROACH: The pharmacokinetic profiles of inhibitors were determined following p.o. (oral) administration and serial bleeding in mice. Subsequently the pharmacokinetics of trans-4-[4-(3-adamantan-1-yl-ureido)-cyclohexyloxy]-benzoic acid (t-AUCB), the most promising inhibitor, was further studied following s.c. (subcutaneous), i.v. (intravenous) injections and administration in drinking water. Finally, the anti-inflammatory effect of t-AUCB was evaluated by using a lipopolysaccharide (LPS)-treated murine model. KEY
RESULTS: Better pharmacokinetic parameters (higher C(max), longer t(1/2) and greater AUC) were obtained from the tested inhibitors, compared with AUDA. Oral bioavailability of t-AUCB (0.1 mg.kg(-1)) was 68 +/- 22% (n = 4), and giving t-AUCB in drinking water is recommended as a feasible, effective and easy route of administration for chronic studies. Finally, t-AUCB (p.o.) reversed the decrease in plasma ratio of lipid epoxides to corresponding diols (a biomarker of soluble epoxide hydrolase inhibition) in lipopolysaccharide-treated mice. The in vivo potency of 1 mg.kg(-1) of t-AUCB (p.o.) was better in this inflammatory model than that of 10 mg.kg(-1) of AUDA-butyl ester (p.o) at 6 h after treatment. CONCLUSIONS AND IMPLICATIONS: t-AUCB is a potent sEHI with improved pharmacokinetic properties. This compound will be a useful tool for pharmacological research and a promising starting point for drug development.
ESTHER : Liu_2009_Br.J.Pharmacol_156_284
PubMedSearch : Liu_2009_Br.J.Pharmacol_156_284
PubMedID: 19154430

Title : Inhibition of the soluble epoxide hydrolase by tyrosine nitration - Barbosa-Sicard_2009_J.Biol.Chem_284_28156
Author(s) : Barbosa-Sicard E , Fromel T , Keseru B , Brandes RP , Morisseau C , Hammock BD , Braun T , Kruger M , Fleming I
Ref : Journal of Biological Chemistry , 284 :28156 , 2009
Abstract : Inhibition of the soluble epoxide hydrolase (sEH) has beneficial effects on vascular inflammation and hypertension indicating that the enzyme may be a promising target for drug development. As the enzymatic core of the hydrolase domain of the human sEH contains two tyrosine residues (Tyr(383) and Tyr(466)) that are theoretically crucial for enzymatic activity, we addressed the hypothesis that the activity of the sEH may be affected by nitrosative stress. Epoxide hydrolase activity was detected in human and murine endothelial cells as well in HEK293 cells and could be inhibited by either authentic peroxynitrite (ONOO(-)) or the ONOO(-) generator 3-morpholino-sydnonimine (SIN-1). Protection of the enzymatic core with 1-adamantyl-3-cyclohexylurea in vitro decreased sensitivity to SIN-1. Both ONOO(-) and SIN-1 elicited the tyrosine nitration of the sEH protein and mass spectrometry analysis of tryptic fragments revealed nitration on several tyrosine residues including Tyr(383) and Tyr(466). Mutation of the latter residues to phenylalanine was sufficient to abrogate epoxide hydrolase activity. In vivo, streptozotocin-induced diabetes resulted in the tyrosine nitration of the sEH in murine lungs and a significant decrease in its activity. Taken together, these data indicate that the activity of the sEH can be regulated by the tyrosine nitration of the protein. Moreover, nitrosative stress would be expected to potentiate the physiological actions of arachidonic acid epoxides by preventing their metabolism to the corresponding diols.
ESTHER : Barbosa-Sicard_2009_J.Biol.Chem_284_28156
PubMedSearch : Barbosa-Sicard_2009_J.Biol.Chem_284_28156
PubMedID: 19704161

Title : 14,15-Epoxyeicosa-5,8,11-trienoic acid (14,15-EET) surrogates containing epoxide bioisosteres: influence upon vascular relaxation and soluble epoxide hydrolase inhibition - Falck_2009_J.Med.Chem_52_5069
Author(s) : Falck JR , Kodela R , Manne R , Atcha KR , Puli N , Dubasi N , Manthati VL , Capdevila JH , Yi XY , Goldman DH , Morisseau C , Hammock BD , Campbell WB
Ref : Journal of Medicinal Chemistry , 52 :5069 , 2009
Abstract : All-cis-14,15-epoxyeicosa-5,8,11-trienoic acid (14,15-EET) is a labile, vasodilatory eicosanoid generated from arachidonic acid by cytochrome P450 epoxygenases. A series of robust, partially saturated analogues containing epoxide bioisosteres were synthesized and evaluated for relaxation of precontracted bovine coronary artery rings and for in vitro inhibition of soluble epoxide hydrolase (sEH). Depending upon the bioisostere and its position along the carbon chain, varying levels of vascular relaxation and/or sEH inhibition were observed. For example, oxamide 16 and N-iPr-amide 20 were comparable (ED(50) 1.7 microM) to 14,15-EET as vasorelaxants but were approximately 10-35 times less potent as sEH inhibitors (IC(50) 59 and 19 microM, respectively); unsubstituted urea 12 showed useful activity in both assays (ED(50) 3.5 microM, IC(50) 16 nM). These data reveal differential structural parameters for the two pharmacophores that could assist the development of potent and specific in vivo drug candidates.
ESTHER : Falck_2009_J.Med.Chem_52_5069
PubMedSearch : Falck_2009_J.Med.Chem_52_5069
PubMedID: 19653681

Title : Differential subcellular distribution and colocalization of the microsomal and soluble epoxide hydrolases in cultured neonatal rat brain cortical astrocytes - Rawal_2009_J.Neurosci.Res_87_218
Author(s) : Rawal S , Morisseau C , Hammock BD , Shivachar AC
Ref : Journal of Neuroscience Research , 87 :218 , 2009
Abstract : The microsomal epoxide hydrolase (mEH) and soluble epoxide hydrolase (sEH) enzymes exist in a variety of cells and tissues, including liver, kidney, and testis. However, very little is known about brain epoxide hydrolases. Here we report the expression, localization, and subcellular distribution of mEH and sEH in cultured neonatal rat cortical astrocytes by immunocytochemistry, subcellular fractionation, Western blotting, and radiometric enzyme assays. Our results showed a diffuse immunofluorescence pattern for mEH, which colocalized with the astroglial cytoskeletal marker glial fibrillary acidic protein (GFAP). The GFAP-positive cells also expressed sEH, which was localized mainly in the cytoplasm, especially in and around the nucleus. Western blot analyses revealed a distinct protein band with a molecular mass of approximately 50 kDa, the signal intensity of which increased about 1.5-fold in the microsomal fraction over the whole-cell lysate and other subcellular fractions. The polyclonal anti-human sEH rabbit serum recognized a protein band with a molecular mass similar to that of the affinity-purified sEH protein (approximately 62 kDa), the signal intensity of which increased over 1.7-fold in the 105,000g supernatant fraction over the cell lysate. Furthermore, the corresponding enzyme activities measured by using mEH- and sEH-selective substrates generally corroborated the immunocytochemical and Western blotting data. These results suggest that rat brain cortical astrocytes differentially coexpress mEH and sEH enzymes. The differential subcellular localization of mEH and sEH may play a role in the cerebrovascular functions that are known to be affected by brain-derived vasoactive epoxides.
ESTHER : Rawal_2009_J.Neurosci.Res_87_218
PubMedSearch : Rawal_2009_J.Neurosci.Res_87_218
PubMedID: 18711743

Title : Sorafenib has soluble epoxide hydrolase inhibitory activity, which contributes to its effect profile in vivo - Liu_2009_Mol.Cancer.Ther_8_2193
Author(s) : Liu JY , Park SH , Morisseau C , Hwang SH , Hammock BD , Weiss RH
Ref : Mol Cancer Ther , 8 :2193 , 2009
Abstract : The advent of multikinase inhibitors targeting the vascular endothelial growth factor (VEGF) receptor has revolutionized the treatment of highly angiogenic malignancies such as renal cell carcinoma. Interestingly, several such inhibitors are commercially available, and they each possess diverse specific beneficial and adverse effect profiles. In examining the structure of sorafenib, it was hypothesized that this compound would possess inhibitory effects on the soluble epoxide hydrolase, an enzyme with pleiotropic effects on inflammation and vascular disease. We now show that sorafenib but not another VEGF receptor targeted inhibitor sunitinib is a potent inhibitor of the human soluble epoxide hydrolase in vitro (K(I) = 17 +/- 4 nmol/L). Furthermore, sorafenib causes the expected in vivo shift in oxylipid profile resulting from soluble epoxide hydrolase inhibition, evidence of a reduction in the acute inflammatory response. Lipopolysaccharide-induced hypotension was reversed with sorafenib but not sunitinib treatment, suggesting that soluble epoxide hydrolase inhibition accounts for at least part of the anti-inflammatory effect of sorafenib. The pharmacokinetic studies presented here in light of the known potency of sorafenib as a soluble epoxide hydrolase inhibitor indicate that the soluble epoxide hydrolase will be largely inhibited at therapeutic doses of sorafenib. Thus, it is likely that soluble epoxide hydrolase inhibition contributes to the beneficial effects from the inhibition of the VEGF receptor and other kinases during treatment with sorafenib.
ESTHER : Liu_2009_Mol.Cancer.Ther_8_2193
PubMedSearch : Liu_2009_Mol.Cancer.Ther_8_2193
PubMedID: 19671760

Title : Development of metabolically stable inhibitors of Mammalian microsomal epoxide hydrolase - Morisseau_2008_Chem.Res.Toxicol_21_951
Author(s) : Morisseau C , Newman JW , Wheelock CE , Hill Iii T , Morin D , Buckpitt AR , Hammock BD
Ref : Chemical Research in Toxicology , 21 :951 , 2008
Abstract : The microsomal epoxide hydrolase (mEH) plays a significant role in the metabolism of xenobiotics such as polyaromatic toxicants. Additionally, polymorphism studies have underlined a potential role of this enzyme in relation to a number of diseases, such as emphysema, spontaneous abortion, eclampsia, and several forms of cancer. We recently demonstrated that fatty amides, such as elaidamide, represent a new class of potent inhibitors of mEH. While these compounds are very active on recombinant mEH in vitro, they are quickly inactivated in liver extracts reducing their value in vivo. We investigated the effect of structural changes on mEH inhibition potency and microsomal stability. Results obtained indicate that the presence of a small alkyl group alpha to the terminal amide function and a thio-ether beta to this function increased mEH inhibition by an order of magnitude while significantly reducing microsomal inactivation. The addition of a hydroxyl group 9-10 carbons from the terminal amide function resulted in better inhibition potency without improving microsomal stability. The best compound obtained, 2-nonylsulfanyl-propionamide, is a competitive inhibitor of mEH with a K I of 72 nM. Furthermore, this new inhibitor significantly reduces mEH diol production in ex vivo lungs exposed to naphthalene, underlying the usefulness of the inhibitors described herein. These novel inhibitors could be valuable tools to investigate the physiological and biological roles of mEH.
ESTHER : Morisseau_2008_Chem.Res.Toxicol_21_951
PubMedSearch : Morisseau_2008_Chem.Res.Toxicol_21_951
PubMedID: 18363382
Gene_locus related to this paper: human-EPHX1 , ratno-hyep

Title : Soluble epoxide hydrolase inhibitors reduce the development of atherosclerosis in apolipoprotein e-knockout mouse model - Ulu_2008_J.Cardiovasc.Pharmacol_52_314
Author(s) : Ulu A , Davis BB , Tsai HJ , Kim IH , Morisseau C , Inceoglu B , Fiehn O , Hammock BD , Weiss RH
Ref : J Cardiovasc Pharmacol , 52 :314 , 2008
Abstract : To determine whether sEH inhibitors influence atherosclerotic lesion formation, we used an established murine model of accelerated atherogenesis, ApoE knockout (-/-) mice. The sEH inhibitor, 1-adamantan-3-(5-(2-(2-ethylethoxy)ethoxy)pentyl)urea (AEPU) was delivered in drinking water. All animals were fed an atherogenic diet while simultaneously infused with angiotensin II by osmotic minipump to induce atherosclerosis. In AEPU-treated animals, there was a 53% reduction in atherosclerotic lesions in the descending aortae as compared to control aortae. AEPU and its major metabolites were detected in the plasma of animals which received it. As expected from the inhibition of sEH, a significant increase in linoleic and arachidonic acid epoxides, as well as an increase in individual 11,12-EET/DHET and 14,15-EET/DHET ratios, were observed. The reduction in atherosclerotic lesion area was inversely correlated with 11,12- and 14,15- EET/DHET ratios, suggesting that the reduction corresponds to the inhibition of sEH. Our data suggest that orally-available sEH inhibitors may be useful in the treatment of patients with atherosclerotic cardiovascular disease.
ESTHER : Ulu_2008_J.Cardiovasc.Pharmacol_52_314
PubMedSearch : Ulu_2008_J.Cardiovasc.Pharmacol_52_314
PubMedID: 18791465

Title : Inhibition of soluble epoxide hydrolase does not protect against endotoxin-mediated hepatic inflammation - Fife_2008_J.Pharmacol.Exp.Ther_327_707
Author(s) : Fife KL , Liu Y , Schmelzer KR , Tsai HJ , Kim IH , Morisseau C , Hammock BD , Kroetz DL
Ref : Journal of Pharmacology & Experimental Therapeutics , 327 :707 , 2008
Abstract : Epoxyeicosatrienoic acids (EETs) are derived from cytochrome P450-catalyzed epoxygenation of arachidonic acid and have emerged as important mediators of numerous biological effects. The major elimination pathway for EETs is through soluble epoxide hydrolase (sEH)-catalyzed metabolism to dihydroxyeicosatrienoic acids (DHETs). Based on previous studies showing that EETs have anti-inflammatory effects, we hypothesized that chronic inhibition of sEH would attenuate a lipopolysaccharide (LPS)-induced inflammatory response in vivo. Continuous dosing of the sEH inhibitors 12-(3-adamantan-1-ylureido)-dodecanoic acid (AUDA), a polyethylene glycol ester of AUDA, and 1-adamantan-1-yl-3-(5-(2-(2-ethoxyethoxy)ethoxy)-pentyl)urea resulted in robust exposure to the inhibitor and target engagement, as evidenced by significant increases in plasma EET/DHET ratios following 6 days of inhibitor treatment. However, sEH inhibitor treatment was not associated with an attenuation of LPS-induced inflammatory gene expression in the liver, and AUDA did not protect from LPS-induced neutrophil infiltration. Furthermore, Ephx2-/-mice that lack sEH expression and have significantly increased plasma EET/DHET ratios were not protected from LPS-induced inflammatory gene expression or neutrophil accumulation in the liver. LPS did have an effect on sEH expression and function, as evident from a significant down-regulation of Ephx2 mRNA and a significant shift in plasma EET/DHET ratios 4 h after LPS treatment. In conclusion, there was no evidence that increasing EET levels in vivo could modulate an LPS-induced inflammatory response in the liver. However, LPS did have significant effects on plasma eicosanoid levels and hepatic Ephx2 expression, suggesting that in vivo EET levels are modulated in response to an inflammatory signal.
ESTHER : Fife_2008_J.Pharmacol.Exp.Ther_327_707
PubMedSearch : Fife_2008_J.Pharmacol.Exp.Ther_327_707
PubMedID: 18815352

Title : Transcriptional regulation of the human soluble epoxide hydrolase gene EPHX2 - Tanaka_2008_Biochim.Biophys.Acta_1779_17
Author(s) : Tanaka H , Kamita SG , Wolf NM , Harris TR , Wu Z , Morisseau C , Hammock BD
Ref : Biochimica & Biophysica Acta , 1779 :17 , 2008
Abstract : Soluble epoxide hydrolase (sEH) is a multifunctional protein encoded by the EPHX2 gene. The biological functions and enzyme kinetics of sEH have been extensively investigated, however, little is known about its transcriptional regulation and mechanisms of tissue specific expression. Here, a luciferase gene based reporter assay was used to identify the minimal promoter and cis regulatory elements of EPHX2. The minimal promoter was identified as a GC-rich region between nts -374 and +28 with respect to the putative transcriptional start site. A reporter plasmid carrying this minimal promoter showed higher or similar activities in comparison to a reporter plasmid carrying nts -5,974 to +28 of EPHX2 in 9 human cell lines that were tested. Sp1 binding sites that are involved in augmenting the minimal promoter activity of EPHX2 were identified by nested deletion analysis, site-specific mutation, electrophoretic mobility shift assay, and chromatin immunoprecipitation assay.
ESTHER : Tanaka_2008_Biochim.Biophys.Acta_1779_17
PubMedSearch : Tanaka_2008_Biochim.Biophys.Acta_1779_17
PubMedID: 18078836

Title : Hydrolysis of cis- and trans-epoxyeicosatrienoic acids by rat red blood cells - Jiang_2008_J.Pharmacol.Exp.Ther_326_330
Author(s) : Jiang H , Zhu AG , Mamczur M , Morisseau C , Hammock BD , Falck JR , McGiff JC
Ref : Journal of Pharmacology & Experimental Therapeutics , 326 :330 , 2008
Abstract : Erythrocytes serve as reservoirs for cis- and trans-epoxyeicosatrienoic acids (EETs). Incubation of rat red blood cells (RBCs) with cis- and trans-EETs produces threo- and erythro-dihydroxyeicosatrienoic acids, respectively. The V(max) of EET hydrolysis by rat intact RBCs (2.35 +/- 0.24 pmol/min/10(8) RBCs for 14,15-trans-EET) decreased by approximately 2 to 3-fold sequentially from 14,15-, 11,12- to 8,9-EETs for both cis- and trans-isomers. The V(max) of trans-EET hydrolysis by RBCs is approximately 2 to 3 times that of the corresponding cis-EETs. Incubation of EETs with recombinant murine soluble epoxide hydrolase (sEH) yielded the same geometric and regio preferences of EET hydrolysis as with rat intact RBCs. The principal epoxide hydrolase activity for EET hydrolysis (approximately 90%) is present in the erythrocyte cytosol. Western blots of sEH suggested a concentration of sEH protein to be approximately 2 microg/mg protein or 0.4 microg/10(9) RBCs. The apparent K(m) values of EETs were between 1 and 2 microM, close to the K(m) for purified sEH as reported. Erythrocyte hydration of cis- and trans-EETs was blocked by sEH inhibitors, 1,3-dicyclohexylurea and 4-[4-(3-adamantan-1-ylureido)cyclohexyloxy]benzoic acid. Erythrocyte sEH activity was inhibited more than 80% by 0.2% bovine serum albumin in the buffer. Preferred hydrolysis of 14,15-EETs and trans-epoxides characterizes sEH activity in RBCs that regulates the hydrolysis and release of cis- and trans-EETs in the circulation. Inhibition of sEH has produced antihypertensive and antiinflammatory effects. Because plasma trans-EETs would increase more than cis-EETs with sEH inhibition, the potential roles of trans-EETs and erythrocyte sEH in terms of circulatory regulation deserve attention.
ESTHER : Jiang_2008_J.Pharmacol.Exp.Ther_326_330
PubMedSearch : Jiang_2008_J.Pharmacol.Exp.Ther_326_330
PubMedID: 18445784

Title : Gerry Brooks and epoxide hydrolases: four decades to a pharmaceutical - Morisseau_2008_Pest.Manag.Sci_64_594
Author(s) : Morisseau C , Hammock BD
Ref : Pest Manag Sci , 64 :594 , 2008
Abstract : The pioneering work of Gerry Brooks on cyclodiene insecticides led to the discovery of a class of enzymes known as epoxide hydrolases. The results from four decades of work confirm Brooks' first observations that the microsomal epoxide hydrolase is important in foreign compound metabolism. Brooks and associates went on to be the first to carry out a systematic study of the inhibition of this enzyme. A second role for this enzyme family was in the degradation of insect juvenile hormone (JH). JH epoxide hydrolases have now been cloned and expressed from several species, and there is interest in developing inhibitors for them. Interestingly, the distantly related mammalian soluble epoxide hydrolase has emerged as a promising pharmacological target for treating hypertension, inflammatory disease and pain. Tight-binding transition-state inhibitors were developed with good ADME (absorption, distribution, metabolism and excretion). These compounds stabilize endogenous epoxides of fatty acids, including arachidonic acid, which have profound therapeutic effects. Now EHs from microorganisms and plants are used in green chemistry. From his seminal work, Dr Brooks opened the field of epoxide hydrolase research in many directions including xenobiotic metabolism, insect physiology and human health, as well as asymmetric organic synthesis.
ESTHER : Morisseau_2008_Pest.Manag.Sci_64_594
PubMedSearch : Morisseau_2008_Pest.Manag.Sci_64_594
PubMedID: 18383502

Title : The molecular structure of epoxide hydrolase B from Mycobacterium tuberculosis and its complex with a urea-based inhibitor - Biswal_2008_J.Mol.Biol_381_897
Author(s) : Biswal BK , Morisseau C , Garen G , Cherney MM , Garen C , Niu C , Hammock BD , James MN
Ref : Journal of Molecular Biology , 381 :897 , 2008
Abstract : Mycobacterium tuberculosis (Mtb), the intracellular pathogen that infects macrophages primarily, is the causative agent of the infectious disease tuberculosis in humans. The Mtb genome encodes at least six epoxide hydrolases (EHs A to F). EHs convert epoxides to trans-dihydrodiols and have roles in drug metabolism as well as in the processing of signaling molecules. Herein, we report the crystal structures of unbound Mtb EHB and Mtb EHB bound to a potent, low-nanomolar (IC(50) approximately 19 nM) urea-based inhibitor at 2.1 and 2.4 A resolution, respectively. The enzyme is a homodimer; each monomer adopts the classical alpha/beta hydrolase fold that composes the catalytic domain; there is a cap domain that regulates access to the active site. The catalytic triad, comprising Asp104, His333 and Asp302, protrudes from the catalytic domain into the substrate binding cavity between the two domains. The urea portion of the inhibitor is bound in the catalytic cavity, mimicking, in part, the substrate binding; the two urea nitrogen atoms donate hydrogen bonds to the nucleophilic carboxylate of Asp104, and the carbonyl oxygen of the urea moiety receives hydrogen bonds from the phenolic oxygen atoms of Tyr164 and Tyr272. The phenolic oxygen groups of these two residues provide electrophilic assistance during the epoxide hydrolytic cleavage. Upon inhibitor binding, the binding-site residues undergo subtle structural rearrangement. In particular, the side chain of Ile137 exhibits a rotation of around 120 degrees about its C(alpha)-C(beta) bond in order to accommodate the inhibitor. These findings have not only shed light on the enzyme mechanism but also have opened a path for the development of potent inhibitors with good pharmacokinetic profiles against all Mtb EHs of the alpha/beta type.
ESTHER : Biswal_2008_J.Mol.Biol_381_897
PubMedSearch : Biswal_2008_J.Mol.Biol_381_897
PubMedID: 18585390
Gene_locus related to this paper: myctu-ephB

Title : 1,3-disubstituted ureas functionalized with ether groups are potent inhibitors of the soluble epoxide hydrolase with improved pharmacokinetic properties - Kim_2007_J.Med.Chem_50_5217
Author(s) : Kim IH , Tsai HJ , Nishi K , Kasagami T , Morisseau C , Hammock BD
Ref : Journal of Medicinal Chemistry , 50 :5217 , 2007
Abstract : Soluble epoxide hydrolase (sEH) is a therapeutic target for treating hypertension and inflammation. 1,3-Disubstituted ureas functionalized with an ether group are potent sEH inhibitors. However, their relatively low metabolic stability leads to poor pharmacokinetic properties. To improve their bioavailability, we investigated the effect of incorporating various polar groups on the ether function on the inhibition potencies, physical properties, in vitro metabolic stability, and pharmacokinetic properties. The structure-activity relationship studies showed that a hydrophobic linker between the urea group and the ether function is necessary to keep their potency. In addition, urea-ether inhibitors having a polar group such as diethylene glycol or morpholine significantly improved their physical properties and metabolic stability without any loss of inhibitory potency. Furthermore, improved pharmacokinetic properties in murine and canine models were obtained with the resulting inhibitors. These findings will facilitate the usage of sEH inhibitors in animal models of hypertension and inflammation.
ESTHER : Kim_2007_J.Med.Chem_50_5217
PubMedSearch : Kim_2007_J.Med.Chem_50_5217
PubMedID: 17894481

Title : Orally bioavailable potent soluble epoxide hydrolase inhibitors - Hwang_2007_J.Med.Chem_50_3825
Author(s) : Hwang SH , Tsai HJ , Liu JY , Morisseau C , Hammock BD
Ref : Journal of Medicinal Chemistry , 50 :3825 , 2007
Abstract : A series of N,N'-disubstituted ureas having a conformationally restricted cis- or trans-1,4-cyclohexane alpha to the urea were prepared and tested as soluble epoxide hydrolase (sEH) inhibitors. This series of compounds showed low nanomolar to picomolar activities against recombinant human sEH. Both isomers showed similar potencies, but the trans isomers were more metabolically stable in human hepatic microsomes. Furthermore, these new potent inhibitors show a greater metabolic stability in vivo than previously described sEH inhibitors. We demonstrated that trans-4-[4-(3-adamantan-1-ylureido)cyclohexyloxy]benzoic acid 13g (t-AUCB, IC50 = 1.3 +/- 0.05 nM) had excellent oral bioavailability (98%, n = 2) and blood area under the curve in dogs and was effective in vivo to treat hypotension in lipopolysaccharide challenged murine models.
ESTHER : Hwang_2007_J.Med.Chem_50_3825
PubMedSearch : Hwang_2007_J.Med.Chem_50_3825
PubMedID: 17616115

Title : Design of bioavailable derivatives of 12-(3-adamantan-1-yl-ureido)dodecanoic acid, a potent inhibitor of the soluble epoxide hydrolase - Kim_2007_Bioorg.Med.Chem_15_312
Author(s) : Kim IH , Nishi K , Tsai HJ , Bradford T , Koda Y , Watanabe T , Morisseau C , Blanchfield J , Toth I , Hammock BD
Ref : Bioorganic & Medicinal Chemistry , 15 :312 , 2007
Abstract : The soluble epoxide hydrolase (sEH) plays an important role in the metabolism of endogenous chemical mediators involved in blood pressure regulation and vascular inflammation. 12-(3-Adamantan-1-yl-ureido)-dodecanoic acid (AUDA, 1) is a very active inhibitor of sEH both in vitro and in vivo. However, its relatively high melting point and limited solubility in either water or oil-based solvents leads to difficulties in formulating the compound and often results in poor in vivo availability. We investigated the effect of derivatization of the acid functional group of inhibitor 1 on the inhibition potencies, physical properties, and pharmacokinetic properties. For human sEH, similar inhibition potency was obtained when the acid of compound 1 was modified to esters (2-15). The resulting compounds exhibited improved physical properties (23-66 degrees C lower melting point and 5-fold better solubility in oil). Pharmacokinetic studies showed that the esters possess improved oral bioavailability in mice. On the other hand, amide derivatives of AUDA 1 did not show significant improvement in inhibition potencies or physical properties (higher melting points and lower solubility). The esterification of 1 results in compounds that are easier to formulate in animal food and in triglycerides for gavage and other routes of administration, making it easier to study the biological effects of sEH inhibition in vivo.
ESTHER : Kim_2007_Bioorg.Med.Chem_15_312
PubMedSearch : Kim_2007_Bioorg.Med.Chem_15_312
PubMedID: 17046265
Gene_locus related to this paper: human-EPHX2

Title : Soluble epoxide hydrolase: a novel therapeutic target in stroke - Zhang_2007_J.Cereb.Blood.Flow.Metab_27_1931
Author(s) : Zhang W , Koerner IP , Noppens R , Grafe M , Tsai HJ , Morisseau C , Luria A , Hammock BD , Falck JR , Alkayed NJ
Ref : Journal of Cerebral Blood Flow & Metabolism , 27 :1931 , 2007
Abstract : The P450 eicosanoids epoxyeicosatrienoic acids (EETs) are produced in brain and perform important biological functions, including protection from ischemic injury. The beneficial effect of EETs, however, is limited by their metabolism via soluble epoxide hydrolase (sEH). We tested the hypothesis that sEH inhibition is protective against ischemic brain damage in vivo by a mechanism linked to enhanced cerebral blood flow (CBF). We determined expression and distribution of sEH immunoreactivity (IR) in brain, and examined the effect of sEH inhibitor 12-(3-adamantan-1-yl-ureido)-dodecanoic acid butyl ester (AUDA-BE) on CBF and infarct size after experimental stroke in mice. Mice were administered a single intraperitoneal injection of AUDA-BE (10 mg/kg) or vehicle at 30 mins before 2-h middle cerebral artery occlusion (MCAO) or at reperfusion, in the presence and absence of P450 epoxygenase inhibitor N-methylsulfonyl-6-(2-propargyloxyphenyl) hexanamide (MS-PPOH). Immunoreactivity for sEH was detected in vascular and non-vascular brain compartments, with predominant expression in neuronal cell bodies and processes. 12-(3-Adamantan-1-yl-ureido)-dodecanoic acid butyl ester was detected in plasma and brain for up to 24 h after intraperitoneal injection, which was associated with inhibition of sEH activity in brain tissue. Finally, AUDA-BE significantly reduced infarct size at 24 h after MCAO, which was prevented by MS-PPOH. However, regional CBF rates measured by iodoantipyrine (IAP) autoradiography at end ischemia revealed no differences between AUDA-BE- and vehicle-treated mice. The findings suggest that sEH inhibition is protective against ischemic injury by non-vascular mechanisms, and that sEH may serve as a therapeutic target in stroke.
ESTHER : Zhang_2007_J.Cereb.Blood.Flow.Metab_27_1931
PubMedSearch : Zhang_2007_J.Cereb.Blood.Flow.Metab_27_1931
PubMedID: 17440491
Gene_locus related to this paper: human-EPHX2

Title : Characterization of pyrethroid hydrolysis by the human liver carboxylesterases hCE-1 and hCE-2 - Nishi_2006_Arch.Biochem.Biophys_445_115
Author(s) : Nishi K , Huang H , Kamita SG , Kim IH , Morisseau C , Hammock BD
Ref : Archives of Biochemistry & Biophysics , 445 :115 , 2006
Abstract : Carboxylesterases hydrolyze a large array of endogenous and exogenous ester-containing compounds, including pyrethroid insecticides. Herein, we report the specific activities and kinetic parameters of human carboxylesterase (hCE)-1 and hCE-2 using authentic pyrethroids and pyrethroid-like, fluorescent surrogates. Both hCE-1 and hCE-2 hydrolyzed type I and II pyrethroids with strong stereoselectivity. For example, the trans-isomers of permethrin and cypermethrin were hydrolyzed much faster than corresponding cis-counterparts by both enzymes. Kinetic values of hCE-1 and hCE-2 were determined using cypermethrin and 11 stereoisomers of the pyrethroid-like, fluorescent surrogates. K(m) values for the authentic pyrethroids and fluorescent surrogates were in general lower than those for other ester-containing substrates of hCEs. The pyrethroid-like, fluorescent surrogates were hydrolyzed at rates similar to the authentic pyrethroids by both enzymes, suggesting the potential of these compounds as tools for high throughput screening of esterases that hydrolyze pyrethroids.
ESTHER : Nishi_2006_Arch.Biochem.Biophys_445_115
PubMedSearch : Nishi_2006_Arch.Biochem.Biophys_445_115
PubMedID: 16359636
Gene_locus related to this paper: human-CES1 , human-CES2

Title : Substituted adamantyl-urea inhibitors of the soluble epoxide hydrolase dilate mesenteric resistance vessels - Olearczyk_2006_J.Pharmacol.Exp.Ther_318_1307
Author(s) : Olearczyk JJ , Field MB , Kim IH , Morisseau C , Hammock BD , Imig JD
Ref : Journal of Pharmacology & Experimental Therapeutics , 318 :1307 , 2006
Abstract : The epoxyeicosatrienoic acids (EETs) have been identified as endothelium-derived hyperpolarizing factors. Metabolism of the EETs to the dihydroxyeicosatrienoic acids is catalyzed by soluble epoxide hydrolase (sEH). Administration of urea-based sEH inhibitors provides protection from hypertension-induced renal injury at least in part by lowering blood pressure. Here, we investigated the hypothesis that a mechanism by which sEH inhibitors elicit their cardiovascular protective effects is via their action on the vasculature. Mesenteric resistance arteries were isolated from Sprague-Dawley rats, pressurized, and constricted with the thromboxane A2 agonist U46619 (9,11-dideoxy-11,9-epoxymethano-prostaglandin F2alpha). Mesenteric arteries were then incubated with increasing concentrations of the sEH inhibitor 12-(3-adamantan-1-yl-ureido)dodecanoic acid (AUDA). AUDA resulted in a concentration-dependent relaxation of mesenteric arteries, with 10 microM resulting in a 48 +/- 7% relaxation. Chain-shortened analogs of AUDA had an attenuated vasodilatory response. Interestingly, at 10 microM, the sEH inhibitors 1-cyclohexyl-3-dodecylurea, 12-(3-cyclohexylureido)dodecanoic acid, and 950 [adamantan-1-yl-3-{5-[2-(2-ethoxyethoxy)ethoxy]pentyl}urea] were significantly less active, resulting in a 25 +/- 8%, 10 +/- 9%, and -8 +/- 3% relaxation, respectively. Treatment of mesenteric arteries with tetraethylammonium, iberiotoxin, ouabain, or glibenclamide did not alter AUDA-induced relaxation. The AUDA-induced relaxation was completely inhibited when constricted with KCl. In separate experiments, denuding mesenteric resistance vessels did not alter AUDA-induced relaxation. Taken together, these data demonstrate that adamantyl-urea inhibitors have unique dilator actions on vascular smooth muscle compared with other sEH inhibitors and that these dilator actions depend on the adamantyl group and carbon chain length.
ESTHER : Olearczyk_2006_J.Pharmacol.Exp.Ther_318_1307
PubMedSearch : Olearczyk_2006_J.Pharmacol.Exp.Ther_318_1307
PubMedID: 16772540
Gene_locus related to this paper: human-EPHX2

Title : Human soluble epoxide hydrolase: structural basis of inhibition by 4-(3-cyclohexylureido)-carboxylic acids - Gomez_2006_Protein.Sci_15_58
Author(s) : Gomez GA , Morisseau C , Hammock BD , Christianson DW
Ref : Protein Science , 15 :58 , 2006
Abstract : X-ray crystal structures of human soluble epoxide hydrolase (sEH) complexed with four different dialkylurea inhibitors bearing pendant carboxylate "tails" of varying length have been determined at 2.3-3.0 A resolution. Similarities among inhibitor binding modes reinforce the proposed roles of Y381 and/or Y465 as general acids that protonate the epoxide ring of the substrate in concert with nucleophilic attack of D333 at the electrophilic epoxide carbon. Additionally, the binding of these inhibitors allows us to model the binding mode of the endogenous substrate 14,15-epoxyeicosatrienoic acid. Contrasts among inhibitor binding modes include opposite orientations of inhibitor binding in the active-site hydrophobic tunnel. Alternative binding orientations observed for this series of inhibitors to human sEH, as well as the binding of certain dialkylurea inhibitors to human sEH and murine sEH, complicate the structure-based design of human sEH inhibitors with potential pharmaceutical applications in the treatment of hypertension. Thus, with regard to the optimization of inhibitor designs targeting human sEH, it is critical that human sEH and not murine sEH be utilized for inhibitor screening, and it is critical that structures of human sEH-inhibitor complexes be determined to verify inhibitor binding orientations that correlate with measured affinities.
ESTHER : Gomez_2006_Protein.Sci_15_58
PubMedSearch : Gomez_2006_Protein.Sci_15_58
PubMedID: 16322563
Gene_locus related to this paper: human-EPHX2

Title : Development of a high-throughput screen for soluble epoxide hydrolase inhibition - Wolf_2006_Anal.Biochem_355_71
Author(s) : Wolf NM , Morisseau C , Jones PD , Hock B , Hammock BD
Ref : Analytical Biochemistry , 355 :71 , 2006
Abstract : Mammalian soluble epoxide hydrolase (sEH) represents a highly promising new target for drug development. Chemical inhibition of this enzyme in animal models was shown to treat hypertension and vascular inflammation as well as related syndromes. Existing sEH inhibitors are relatively potent and specific. However, the low solubility and relatively fast metabolism of described sEH inhibitors make them less than therapeutically efficient, stating the need for novel inhibitor structures. Therefore, a series of alpha-cyanoester and alpha-cyanocarbonate epoxides were evaluated as potential human sEH (HsEH) substrates for the high-throughput screen (HTS) of compound libraries. (3-Phenyl-oxiranyl)-acetic acid cyano-(6-methoxy-naphthalen-2-yl)-methyl ester (PHOME), which displayed the highest aqueous stability and solubility, was selected for the development of an HTS assay with long incubation times at room temperature. Concentrations of HsEH and PHOME were optimized to ensure assay sensitivity, reliability, and reproducibility. Assay validation, which employed these optimized concentrations, resulted in good accuracy (60-100%) and high precision (<7% relative standard deviation). In addition, an overall Z' value of 0.7 proved the system's robustness and potential for HTS. The developed assay system will be a valuable tool to discover new structures for the therapeutic inhibition of sEH to treat various cardiovascular diseases.
ESTHER : Wolf_2006_Anal.Biochem_355_71
PubMedSearch : Wolf_2006_Anal.Biochem_355_71
PubMedID: 16729954

Title : Epoxide hydrolases: their roles and interactions with lipid metabolism - Newman_2005_Prog.Lipid.Res_44_1
Author(s) : Newman JW , Morisseau C , Hammock BD
Ref : Prog Lipid Res , 44 :1 , 2005
Abstract : The epoxide hydrolases (EHs) are enzymes present in all living organisms, which transform epoxide containing lipids by the addition of water. In plants and animals, many of these lipid substrates have potent biologically activities, such as host defenses, control of development, regulation of inflammation and blood pressure. Thus the EHs have important and diverse biological roles with profound effects on the physiological state of the host organisms. Currently, seven distinct epoxide hydrolase sub-types are recognized in higher organisms. These include the plant soluble EHs, the mammalian soluble epoxide hydrolase, the hepoxilin hydrolase, leukotriene A4 hydrolase, the microsomal epoxide hydrolase, and the insect juvenile hormone epoxide hydrolase. While our understanding of these enzymes has progressed at different rates, here we discuss the current state of knowledge for each of these enzymes, along with a distillation of our current understanding of their endogenous roles. By reviewing the entire enzyme class together, both commonalities and discrepancies in our understanding are highlighted and important directions for future research pertaining to these enzymes are indicated.
ESTHER : Newman_2005_Prog.Lipid.Res_44_1
PubMedSearch : Newman_2005_Prog.Lipid.Res_44_1
PubMedID: 15748653

Title : Optimization of amide-based inhibitors of soluble epoxide hydrolase with improved water solubility - Kim_2005_J.Med.Chem_48_3621
Author(s) : Kim IH , Heirtzler FR , Morisseau C , Nishi K , Tsai HJ , Hammock BD
Ref : Journal of Medicinal Chemistry , 48 :3621 , 2005
Abstract : Soluble epoxide hydrolase (sEH) plays an important role in the metabolism of endogenous chemical mediators involved in the regulation of blood pressure and inflammation. 1,3-Disubstituted ureas with a polar group located on the fifth atom from the carbonyl group of urea function are active inhibitors of sEH both in vitro and in vivo. However, their limited solubility in water and relatively high melting point lead to difficulties in formulating the compounds and poor in vivo efficacy. To improve these physical properties, the effect of structural modification of the urea pharmacophore on the inhibition potencies, water solubilities, octanol/water partition coefficients (log P), and melting points of a series of compounds was evaluated. For murine sEH, no loss of inhibition potency was observed when the urea pharmacophore was modified to an amide function, while for human sEH 2.5-fold decreased inhibition was obtained in the amide compounds. In addition, a NH group on the right side of carbonyl group of the amide pharmacophore substituted with an adamantyl group (such as compound 14) and a methylene carbon present between the adamantyl and amide groups were essential to produce potent inhibition of sEH. The resulting amide inhibitors have 10-30-fold better solubility and lower melting point than the corresponding urea compounds. These findings will facilitate synthesis of sEH inhibitors that are easier to formulate and more bioavailable.
ESTHER : Kim_2005_J.Med.Chem_48_3621
PubMedSearch : Kim_2005_J.Med.Chem_48_3621
PubMedID: 15887969

Title : Identification and characterization of an ovary-selective isoform of epoxide hydrolase - Hennebold_2005_Biol.Reprod_72_968
Author(s) : Hennebold JD , Mah K , Perez W , Vance JE , Stouffer RL , Morisseau C , Hammock BD , Adashi EY
Ref : Biol Reprod , 72 :968 , 2005
Abstract : A novel ovary-selective gene was identified by suppression subtractive hybridization (SSH) that is expressed only during the mouse periovulatory phase of a stimulated estrous cycle. Analysis of the protein encoded by the full-length cDNA revealed that the majority of it, with the exception of the first 44 amino acids, matched soluble epoxide hydrolase (Ephx2, referred to as Ephx2A). By comparing the cDNA sequence of this newly identified variant of soluble epoxide hydrolase (referred to as Ephx2B) with the mouse genome database, an exon was identified that corresponds to its unique 5' cDNA sequence. Through the use of an Ephx2A-specific probe, Northern blot analysis revealed that this mRNA was also expressed in the ovary, with the highest level of expression occurring during the luteal phase of a stimulated estrous cycle. In situ hybridization revealed that Ephx2B mRNA expression was restricted to granulosa cells of preovulatory follicles. Ephx2A mRNA expression, however, was detectable in follicles at different stages of development, as well as in the corpus luteum. Total ovarian epoxide hydrolase activity increased following the induction of follicular development, and remained elevated through the periovulatory and postovulatory stages of a stimulated estrous cycle. The change in enzyme activity paralleled the combined mRNA expression profiles for both Ephx2A and Ephx2B, thus supporting a role for epoxide metabolism in ovarian function.
ESTHER : Hennebold_2005_Biol.Reprod_72_968
PubMedSearch : Hennebold_2005_Biol.Reprod_72_968
PubMedID: 15601917

Title : Novel mechanism of brain soluble epoxide hydrolase-mediated blood pressure regulation in the spontaneously hypertensive rat - Sellers_2005_FASEB.J_19_626
Author(s) : Sellers KW , Sun C , Diez-Freire C , Waki H , Morisseau C , Falck JR , Hammock BD , Paton JF , Raizada MK
Ref : FASEB Journal , 19 :626 , 2005
Abstract : The role of soluble epoxide hydrolase (sEH) in the central control of blood pressure (BP) has not been elucidated in spite of peripheral sEH overexpression being linked to hypertension. Thus, our objective was to investigate the involvement of brain sEH in BP control. sEH expression in the hypothalamus and brain stem, two cardioregulatory brain areas, was increased in the spontaneously hypertensive rat (SHR) compared to the Wistar Kyoto (WKY) rat. Inhibition of the enzyme by intracerebroventricular (icv) delivery of AUDA further increased both BP and heart rate (HR) by 32 +/- 6 mmHg and 54 +/- 10 bpm, respectively, (P<0.05) in the SHR. Analysis of waveform telemetry data revealed a decrease in spontaneous baroreceptor reflex gain following sEH inhibition, indicating the sustained increase in BP may be due to a decrease in baroreceptor reflex function. The hypertensive effect of sEH inhibition is likely a result of an increase in epoxyeicosatrienoic acid (EET)-mediated generation of ROS. This view is supported by the following: 1) Inhibition of EET formation attenuates AUDA-induced increase in BP; 2) delivery of an EET agonist increases BP and HR in the WKY rat, and 3) inhibition of NAD(P)H oxidase by gp91ds-tat prevents AUDA-induced increases in BP and HR. Finally, electrophysiological studies demonstrate that AUDA increased neuronal firing rate exclusively in the SHR, an effect completely abolished by gp91ds-tat. These observations suggest that EETs and sEH inhibition are involved in increasing BP in the SHR. We suggest that an increased expression of sEH is a futile central nervous system response in protection against hypertension.
ESTHER : Sellers_2005_FASEB.J_19_626
PubMedSearch : Sellers_2005_FASEB.J_19_626
PubMedID: 15659536

Title : Epoxide hydrolases: mechanisms, inhibitor designs, and biological roles - Morisseau_2005_Annu.Rev.Pharmacol.Toxicol_45_311
Author(s) : Morisseau C , Hammock BD
Ref : Annual Review of Pharmacology & Toxicology , 45 :311 , 2005
Abstract : Organisms are exposed to epoxide-containing compounds from both exogenous and endogenous sources. In mammals, the hydration of these compounds by various epoxide hydrolases (EHs) can not only regulate their genotoxicity but also, for lipid-derived epoxides, their endogenous roles as chemical mediators. Recent findings suggest that the EHs as a family represent novel drug discovery targets for regulation of blood pressure, inflammation, cancer progression, and the onset of several other diseases. Knowledge of the EH mechanism provides a solid foundation for the rational design of inhibitors, and this review summarizes the current understanding of the catalytic mechanism of the EHs. Although the overall EH mechanism is now known, the molecular basis of substrate selectivity, possible allosteric regulation, and many fine details of the catalytic mechanism remain to be solved. Finally, recent development in the design of EH inhibitors and the EH biological role are discussed.
ESTHER : Morisseau_2005_Annu.Rev.Pharmacol.Toxicol_45_311
PubMedSearch : Morisseau_2005_Annu.Rev.Pharmacol.Toxicol_45_311
PubMedID: 15822179

Title : Structure of human epoxide hydrolase reveals mechanistic inferences on bifunctional catalysis in epoxide and phosphate ester hydrolysis - Gomez_2004_Biochemistry_43_4716
Author(s) : Gomez GA , Morisseau C , Hammock BD , Christianson DW
Ref : Biochemistry , 43 :4716 , 2004
Abstract : The X-ray crystal structure of human soluble epoxide hydrolase (sEH) has been determined at 2.6 A resolution, revealing a domain-swapped quaternary structure identical to that observed for the murine enzyme [Argiriadi, M. A., Morisseau, C., Hammock, B. D., and Christianson, D. W. (1999) Proc. Natl. Acad. Sci. U.S.A. 96, 10637-10642]. As with the murine enzyme, the epoxide hydrolytic mechanism of the human enzyme proceeds through an alkyl-enzyme intermediate with Asp-333 in the C-terminal domain. The structure of the human sEH complex with N-cyclohexyl-N'-(iodophenyl)urea (CIU) has been determined at 2.35 A resolution. Tyr-381 and Tyr-465 donate hydrogen bonds to the alkylurea carbonyl group of CIU, consistent with the proposed roles of these residues as proton donors in the first step of catalysis. The N-terminal domain of mammalian sEH contains a 15 A deep cleft, but its biological function is unclear. Recent experiments demonstrate that the N-terminal domain of human sEH catalyzes the metal-dependent hydrolysis of phosphate esters [Cronin, A., Mowbray, S., Durk, H., Homburg, S., Fleming, I., Fisslthaler, B., Oesch, F., and Arand, M. (2003) Proc. Natl. Acad. Sci. U.S.A. 100, 1552-1557; Newman, J. W., Morisseau, C., Harris, T. R., and Hammock, B. D. (2003) Proc. Natl. Acad. Sci. U.S.A. 100, 1558-1563]. The binding of Mg(2+)-HPO4(2-) to the N-terminal domain of human sEH in its CIU complex reveals structural features relevant to those of the enzyme-substrate complex in the phosphatase reaction.
ESTHER : Gomez_2004_Biochemistry_43_4716
PubMedSearch : Gomez_2004_Biochemistry_43_4716
PubMedID: 15096040
Gene_locus related to this paper: human-EPHX2

Title : Epoxide hydrolases in the rat epididymis: possible roles in xenobiotic and endogenous fatty acid metabolism - DuTeaux_2004_Toxicol.Sci_78_187
Author(s) : DuTeaux SB , Newman JW , Morisseau C , Fairbairn EA , Jelks K , Hammock BD , Miller MG
Ref : Toxicol Sci , 78 :187 , 2004
Abstract : Epoxide hydrolases play an important role in detoxifying epoxides that arise from the metabolism of xenobiotic and endogenous compounds. Both the soluble and microsomal forms of epoxide hydrolase (sEH and mEH, respectively) have been detected in the rat testis. Because of the important role the epididymis plays in sperm maturation and protection, the present study evaluated the presence and activity of these two epoxide hydrolases in the rat epididymis. Using Western blotting, protein bands consistent in size with both mEH and sEH were detected in the caput, corpus, and cauda of the epididymis. The mEH immunoreactive bands in the epididymis ( approximately 50 kDa) were consistent with mEH detected in the liver and kidney. The sEH immunoreactive bands in the epididymis ( approximately 65 kDa) were consistent with a recombinant sEH standard and sEH detected in the liver, kidney, and testis. The presence of mEH and sEH in the epididymis was supported by observations from substrate-based enzyme assays. Results indicated that epididymal mEH can hydrolyze [(3)H]-cis-stilbene oxide to the corresponding diol at levels approximately 9% of the kidney. Epididymal sEH hydrolyzed the substrate [(3)H]-trans-diphenylpropene oxide to the corresponding diol and this activity was inhibited by cyclohexyl-dodecyl urea. Arachidonic acid epoxygenase activity was detected in epididymal S9 fractions, suggesting that fatty acid metabolism by epididymal cytochrome P450s can form epoxides that subsequently become substrates for epididymal sEH. Results from the present study indicate that the epididymis contains at least two active forms of epoxide hydrolase. The role of these enzymes in the detoxification of xenobiotic epoxides is well known, although it is unclear what cellular role they may play in the formation of biologically active metabolites in the epididymis.
ESTHER : DuTeaux_2004_Toxicol.Sci_78_187
PubMedSearch : DuTeaux_2004_Toxicol.Sci_78_187
PubMedID: 14737000

Title : Investigation of the role of a second conserved serine in carboxylesterases via site-directed mutagenesis - Stok_2004_Arch.Biochem.Biophys_430_247
Author(s) : Stok JE , Goloshchapov A , Song C , Wheelock CE , Derbel MB , Morisseau C , Hammock BD
Ref : Archives of Biochemistry & Biophysics , 430 :247 , 2004
Abstract : Carboxylesterases are enzymes that catalyze the hydrolysis of ester and amide moieties. These enzymes have an active site that is composed of a nucleophile (Ser), a base (His), and an acid (Glu) that is commonly known as a catalytic triad. It has previously been observed that the majority of carboxylesterases and lipases contain a second conserved serine in their active site [Proteins, 34 (1999) 184]. To investigate whether this second serine is also involved in the catalytic mechanism, it was mutated to an alanine, a glycine or a cysteine. Site-directed mutagenesis of this conserved serine resulted in a loss of specific activity, in both the S247G and S247A mutants (5- to 15-fold), which was due to a decrease in the rate of catalysis (kcat). Due to the instability of the S247C mutant no reliable data could be attained. A carbamate inhibitor, carbaryl, was then employed to investigate whether this decrease in the kcat was due to the rate of formation of the acyl-enzyme intermediate (k2) or the rate of deacylation (k3). The S247A mutant was found only to alter k2 (2.5-fold decrease), with no effect on k3. Together with information inferred from a human carboxylesterase crystal structure, it was concluded that this serine provides an important structural support for the spatial orientation of the glutamic acid, stabilizing the catalytic triad so that it can perform the hydrolysis.
ESTHER : Stok_2004_Arch.Biochem.Biophys_430_247
PubMedSearch : Stok_2004_Arch.Biochem.Biophys_430_247
PubMedID: 15369824
Gene_locus related to this paper: ratno-Ces1d

Title : Identification, expression, and purification of a pyrethroid-hydrolyzing carboxylesterase from mouse liver microsomes - Stok_2004_J.Biol.Chem_279_29863
Author(s) : Stok JE , Huang H , Jones PD , Wheelock CE , Morisseau C , Hammock BD
Ref : Journal of Biological Chemistry , 279 :29863 , 2004
Abstract : Carboxylesterases are enzymes that catalyze the hydrolysis of a wide range of ester-containing endogenous and xenobiotic compounds. Although the use of pyrethroids is increasing, the specific enzymes involved in the hydrolysis of these insecticides have yet to be identified. A pyrethroid-hydrolyzing enzyme was partially purified from mouse liver microsomes using a fluorescent reporter similar in structure to cypermethrin (Shan, G., and Hammock, B. D. (2001) Anal. Biochem. 299, 54-62 and Wheelock, C. E., Wheelock, A. M., Zhang, R., Stok, J. E., Morisseau, C., Le Valley, S. E., Green, C. E., and Hammock, B. D. (2003) Anal. Biochem. 315, 208-222) and subsequently identified as a carboxylesterase (NCBI accession number BAC36707). The expressed sequence tag was then cloned, expressed in baculovirus, and purified to homogeneity. Kinetic constants for a large number of both type I and type II pyrethroid or pyrethroid-like substrates were determined. This esterase possesses similar kinetic constants for cypermethrin and its fluorescent-surrogate (k(cat) = 0.12 +/- 0.03 versus 0.11 +/- 0.01 s(-1)). Compared with their cis- counterparts, trans-permethrin and cypermethrin were hydrolyzed 22- and 4-fold faster, respectively. Of the four fenvalerate isomers the (2R)(alphaR)-isomer was hydrolyzed at least 1 order of magnitude faster than any other isomer. However, it is unlikely that this enzyme accounts for the total pyrethroid hydrolysis in the microsomes because both isoelectrofocusing and native PAGE indicate the presence of a second region of cypermethrin-metabolizing enzymes. A second carboxylesterase gene (NCBI accession number NM_133960), isolated during a cDNA mouse liver library screening, was also found to hydrolyze pyrethroids. Both these enzymes could be used as preliminary tools in establishing the relative toxicity of new pyrethroids.
ESTHER : Stok_2004_J.Biol.Chem_279_29863
PubMedSearch : Stok_2004_J.Biol.Chem_279_29863
PubMedID: 15123619
Gene_locus related to this paper: mouse-Ces2a , mouse-Ces2e

Title : Vascular localization of soluble epoxide hydrolase in the human kidney - Yu_2004_Am.J.Physiol.Renal.Physiol_286_F720
Author(s) : Yu Z , Davis BB , Morisseau C , Hammock BD , Olson JL , Kroetz DL , Weiss RH
Ref : American Journal of Physiology Renal Physiol , 286 :F720 , 2004
Abstract : Epoxyeicosatrienoic acids are cytochrome P-450 metabolites of arachidonic acid with multiple biological functions, including the regulation of vascular tone, renal tubular transport, cellular proliferation, and inflammation. Epoxyeicosatrienoic acids are converted by soluble epoxide hydrolase into the corresponding dihydroxyeicosatrienoic acids, and epoxyeicosatrienoic acid hydration is regarded as one mechanism whereby their biological effects are eliminated. Previous animal studies indicate that soluble epoxide hydrolase plays an important role in the regulation of renal eicosanoid levels and systemic blood pressure. To begin to elucidate the mechanism of these effects, we determined the cellular localization of soluble epoxide hydrolase in human kidney by examining biopsies taken from patients with a variety of non-end-stage renal diseases, as well as those without known renal disease. Immunohistochemical staining of acetone-fixed kidney biopsy samples revealed that soluble epoxide hydrolase was preferentially expressed in the renal vasculature with relatively low levels in the surrounding tubules. Expression of soluble epoxide hydrolase was evident in renal arteries of varying diameter and was localized mostly in the smooth muscle layers of the arterial wall. Western blot analysis and functional assays confirmed the expression of soluble epoxide hydrolase in the human kidney. There were no obvious differences in soluble epoxide hydrolase expression between normal and diseased human kidney tissue in the samples examined. Our results indicate that soluble epoxide hydrolase is present in the human kidney, being preferentially expressed in the renal vasculature, and support an essential role for this enzyme in renal hemodynamic regulation and its potential utility as a target for therapeutic intervention.
ESTHER : Yu_2004_Am.J.Physiol.Renal.Physiol_286_F720
PubMedSearch : Yu_2004_Am.J.Physiol.Renal.Physiol_286_F720
PubMedID: 14665429

Title : Design, synthesis, and biological activity of 1,3-disubstituted ureas as potent inhibitors of the soluble epoxide hydrolase of increased water solubility - Kim_2004_J.Med.Chem_47_2110
Author(s) : Kim IH , Morisseau C , Watanabe T , Hammock BD
Ref : Journal of Medicinal Chemistry , 47 :2110 , 2004
Abstract : The soluble epoxide hydrolase (sEH) is involved in the metabolism of endogenous chemical mediators that play an important role in blood pressure regulation and inflammation. 1,3-Disubstituted ureas are potent inhibitors of sEH that are active both in vitro and in vivo. However, their poor solubility in either water or lipid reduces their in vivo efficacy and makes them difficult to formulate. To improve these physical properties, the effect of incorporating polar functional groups into one of the alkyl chains was evaluated on their inhibitor potencies, water solubility, octanol/water partition coefficients (log P), and melting points. No loss of inhibition potency was observed when a polar functional group was incorporated at least five atoms ( approximately 7.5 A) from the central urea carbonyl. In addition, the presence of a polar group at least 11 atoms away from the urea carbonyl group for the mouse and human sEHs, respectively, did not alter the inhibitor potency. The resulting compounds have better water solubility and generally lower log P values and melting points than nonfunctionalized liphophilic ureas. These properties will make the compounds more bioavailable and more soluble in either water- or oil-based formulations.
ESTHER : Kim_2004_J.Med.Chem_47_2110
PubMedSearch : Kim_2004_J.Med.Chem_47_2110
PubMedID: 15056008

Title : The soluble epoxide hydrolase encoded by EPXH2 is a bifunctional enzyme with novel lipid phosphate phosphatase activity - Newman_2003_Proc.Natl.Acad.Sci.U.S.A_100_1558
Author(s) : Newman JW , Morisseau C , Harris TR , Hammock BD
Ref : Proc Natl Acad Sci U S A , 100 :1558 , 2003
Abstract : The gene EPXH2 encodes for the soluble epoxide hydrolase (sEH), an enzyme involved in the regulation of cardiovascular and renal physiology containing two distinct domains connected via a proline-rich linker. The C-terminal domain containing the EH catalytic activity has been well studied. In contrast, a function for the N-terminal domain, which has high homology to the haloacid dehalogenase family of phosphatases, has not been definitively reported. In this study we describe the N-terminal domain as a functional phosphatase unaffected by a number of classic phosphatase inhibitors. Assuming a functional association between these catalytic activities, dihydroxy lipid phosphates were rationalized as potential endogenous substrates. A series of phosphorylated hydroxy lipids were therefore synthesized and found to be excellent substrates for the human sEH. The best substrate tested was the monophosphate of dihydroxy stearic acid (threo-910-phosphonoxy-hydroxy-octadecanoic acid) with K(m) = 21 +/- 0.3 microM, V(Max) = 338 +/- 12 nmol x min(-1) x mg(-1), and k(cat) = 0.35 +/- 0.01 s(-1). Therefore dihydroxy lipid phosphates are possible candidates for the endogenous substrates of the sEH N-terminal domain, which would represent a novel branch of fatty acid metabolism with potential signaling functions.
ESTHER : Newman_2003_Proc.Natl.Acad.Sci.U.S.A_100_1558
PubMedSearch : Newman_2003_Proc.Natl.Acad.Sci.U.S.A_100_1558
PubMedID: 12574510
Gene_locus related to this paper: human-EPHX2

Title : Evaluation of alpha-cyanoesters as fluorescent substrates for examining interindividual variation in general and pyrethroid-selective esterases in human liver microsomes - Wheelock_2003_Anal.Biochem_315_208
Author(s) : Wheelock CE , Wheelock AM , Zhang R , Stok JE , Morisseau C , Le Valley SE , Green CE , Hammock BD
Ref : Analytical Biochemistry , 315 :208 , 2003
Abstract : Carboxylesterases hydrolyze many pharmaceuticals and agrochemicals and have broad substrate selectivity, requiring a suite of substrates to measure hydrolytic profiles. To develop new esterase substrates, a series of alpha-cyanoesters that yield fluorescent products upon hydrolysis was evaluated for use in carboxylesterase assays. The use of these substrates as surrogates for Type II pyrethroid hydrolysis was tested. The results suggest that these novel analogs are appropriate for the development of high-throughput assays for pyrethroid hydrolase activity. A set of human liver microsomes was then used to determine the ability of these substrates to report esterase activity across a small population. Results were compared against standard esterase substrates. A number of the esterase substrates showed correlations, demonstrating the broad substrate selectivity of these enzymes. However, for several of the substrates, no correlations in hydrolysis rates were observed, suggesting that multiple carboxylesterase isozymes are responsible for the array of substrate hydrolytic activity. These new substrates were then compared against alpha-naphthyl acetate and 4-methylumbelliferyl acetate for their ability to detect hydrolytic activity in both one- and two-dimensional native electrophoresis gels. Cyano-2-naphthylmethyl butanoate was found to visualize more activity than either commercial substrate. These applications demonstrate the utility of these new substrates as both general and pyrethroid-selective reporters of esterase activity.
ESTHER : Wheelock_2003_Anal.Biochem_315_208
PubMedSearch : Wheelock_2003_Anal.Biochem_315_208
PubMedID: 12689831
Gene_locus related to this paper: mouse-Ces2e

Title : In vitro metabolism of the mammalian soluble epoxide hydrolase inhibitor, 1-cyclohexyl-3-dodecyl-urea - Watanabe_2003_Drug.Metab.Dispos_31_846
Author(s) : Watanabe T , Morisseau C , Newman JW , Hammock BD
Ref : Drug Metabolism & Disposition: The Biological Fate of Chemicals , 31 :846 , 2003
Abstract : The metabolism of the soluble epoxide hydrolase (sEH) inhibitor, 1-cyclohexyl-3-dodecyl-urea (CDU), was studied in rat and human hepatic microsomes. The microsomal metabolism of CDU enhanced sEH inhibition potency of the reaction mixture and resulted in the formation of several metabolites. During the course of this study, a sensitive and specific high-performance liquid chromatography with tandem mass spectrometry analytical method was developed to investigate simultaneously the production of these metabolites. In both rat and human hepatic microsomes, CDU was ultimately transformed into the corresponding omega-carboxylate; however, the rodent tissue appeared to perform this transformation more rapidly. After a 60-min incubation in rat hepatic microsomes, the percentage of residual CDU, the omega-carboxylate, and the intermediary omega-hydroxyl were about 20%, 20%, and 50%, respectively. Carbon monoxide inhibited the metabolism of CDU by rat hepatic microsomes, suggesting that the initial step is catalyzed by cytochrome P450. Further metabolism was enhanced by the addition of NAD, suggesting that dehydrogenases are associated with intermediate metabolic steps. Regardless, the ultimate product of microsomal metabolism, 12-(3-cyclohexyl-ureido)-dodecanoic acid, is also an excellent sEH inhibitor with several hundred-fold higher solubility, supporting the hypothesis that CDU has prodrug characteristics. These findings will facilitate the rational design and optimization of sEH inhibitors with better physical properties and improved metabolic stability.
ESTHER : Watanabe_2003_Drug.Metab.Dispos_31_846
PubMedSearch : Watanabe_2003_Drug.Metab.Dispos_31_846
PubMedID: 12814960

Title : QSAR and classification of murine and human soluble epoxide hydrolase inhibition by urea-like compounds - McElroy_2003_J.Med.Chem_46_1066
Author(s) : McElroy NR , Jurs PC , Morisseau C , Hammock BD
Ref : Journal of Medicinal Chemistry , 46 :1066 , 2003
Abstract : A data set of 348 urea-like compounds that inhibit the soluble epoxide hydrolase enzyme in mice and humans is examined. Compounds having IC(50) values ranging from 0.06 to >500 microM (murine) and 0.10 to >500 microM (human) are categorized as active or inactive for classification, while quantitation is performed on smaller compound subsets ranging from 0.07 to 431 microM (murine) and 0.11 to 490 microM (human). Each compound is represented by calculated structural descriptors that encode topological, geometrical, electronic, and polar surface features. Multiple linear regression (MLR) and computational neural networks (CNNs) are employed for quantitative models. Three classification algorithms, k-nearest neighbor (kNN), linear discriminant analysis (LDA), and radial basis function neural networks (RBFNN), are used to categorize compounds as active or inactive based on selected data split points. Quantitative modeling of human enzyme inhibition results in a nonlinear, five-descriptor model with root-mean-square errors (log units of IC(50) [microM]) of 0.616 (r(2) = 0.66), 0.674 (r(2) = 0.61), and 0.914 (r(2) = 0.33) for training, cross-validation, and prediction sets, respectively. The best classification results for human and murine enzyme inhibition are found using kNN. Human classification rates using a seven-descriptor model for training and prediction sets are 89.1% and 91.4%, respectively. Murine classification rates using a five-descriptor model for training and prediction sets are 91.5% and 88.6%, respectively.
ESTHER : McElroy_2003_J.Med.Chem_46_1066
PubMedSearch : McElroy_2003_J.Med.Chem_46_1066
PubMedID: 12620084

Title : Polymorphism in soluble epoxide hydrolase and blood pressure in spontaneously hypertensive rats - Fornage_2002_Hypertension_40_485
Author(s) : Fornage M , Hinojos CA , Nurowska BW , Boerwinkle E , Hammock BD , Morisseau C , Doris PA
Ref : Hypertension , 40 :485 , 2002
Abstract : We measured soluble epoxide hydrolase (sEH) renal gene expression in prehypertensive (4 to 5 weeks old) spontaneously hypertensive rats of the Heidelberg SP substrain (SHR [Heid]) and when blood pressure levels entered the hypertensive plateau (17 to 18 weeks old) and compared expression with matched Wistar-Kyoto (WKY [Heid]) rats. Less expression of the gene encoding sEH (EPHX2) was observed in SHR (Heid) than in WKY (Heid). Analysis of sEH protein abundance showed a similar difference. However, no correlation between sEH abundance and blood pressure was observed in the F(2) progeny of a parental strain cross. Measurement of protein abundance in SHR and WKY obtained from Charles River confirmed a recent report that abundance of sEH was greater in SHR (CRiv) than WKY (CRiv) strains. Polymorphisms were detected in EPHX2. Resequencing revealed that 2 alleles of EPHX2 exist in these 4 rat strains, differing by 4 single nucleotide polymorphisms, of which 3 produce nonsynonymous amino acid substitutions. The ancestral allele was shared by SHR (Heid) and WKY (CRiv), and the variant allele was shared by WKY (Heid) and SHR (CRiv). Activity of sEH was greater in animals carrying the variant allele. However, inheritance of this allele was not correlated with blood pressure in the F(2) progeny of a cross between SHR (Heid) and WKY (Heid). These data indicate that sequence variation determining functional alterations in EPHX2 is not likely to contribute to blood pressure levels in SHR.
ESTHER : Fornage_2002_Hypertension_40_485
PubMedSearch : Fornage_2002_Hypertension_40_485
PubMedID: 12364351

Title : Structural refinement of inhibitors of urea-based soluble epoxide hydrolases - Morisseau_2002_Biochem.Pharmacol_63_1599
Author(s) : Morisseau C , Goodrow MH , Newman JW , Wheelock CE , Dowdy DL , Hammock BD
Ref : Biochemical Pharmacology , 63 :1599 , 2002
Abstract : The soluble epoxide hydrolase (sEH) is involved in the metabolism of arachidonic, linoleic, and other fatty acid epoxides, endogenous chemical mediators that play an important role in blood pressure regulation and inflammation. 1,3-Disubstituted ureas, carbamates, and amides are new potent and stable inhibitors of sEH. However, the poor solubility of the lead compounds limits their use. Inhibitor structure-activity relationships were investigated to better define the structural requirements for inhibition and to identify points in the molecular topography that could accept polar groups without diminishing inhibition potency. Results indicate that lipophilicity is an important factor controlling inhibitor potency. Polar groups could be incorporated into one of the alkyl groups without loss of activity if they were placed at a sufficient distance from the urea function. The resulting compounds had a 2-fold higher water solubility. These findings will facilitate the rational design and optimization of sEH inhibitors with better physical properties.
ESTHER : Morisseau_2002_Biochem.Pharmacol_63_1599
PubMedSearch : Morisseau_2002_Biochem.Pharmacol_63_1599
PubMedID: 12007563

Title : Inhibitors of soluble epoxide hydrolase attenuate vascular smooth muscle cell proliferation - Davis_2002_Proc.Natl.Acad.Sci.U.S.A_99_2222
Author(s) : Davis BB , Thompson DA , Howard LL , Morisseau C , Hammock BD , Weiss RH
Ref : Proc Natl Acad Sci U S A , 99 :2222 , 2002
Abstract : Atherosclerosis, in its myriad incarnations the foremost killer disease in the industrialized world, is characterized by aberrant proliferation of vascular smooth muscle (VSM) cells in part as a result of the recruitment of inflammatory cells to the blood vessel wall. The epoxyeicosatrienoic acids are synthesized from arachidonic acid in a reaction catalyzed by the cytochrome P450 system and are vasoactive substances. Metabolism of these compounds by epoxide hydrolases results in the formation of compounds that affect the vasculature in a pleiotropic manner. As an outgrowth of our observations that urea inhibitors of the soluble epoxide hydrolase (sEH) reduce blood pressure in spontaneously hypertensive rats as well as the findings of other investigators that these compounds possess antiinflammatory actions, we have examined the effect of sEH inhibitors on VSM cell proliferation. We now show that the sEH inhibitor 1-cyclohexyl-3-dodecyl urea (CDU) inhibits human VSM cell proliferation in a dose-dependent manner and is associated with a decrease in the level of cyclin D1. In addition, cis-epoxyeicosatrienoic acid mimics the growth-suppressive activity of CDU; there is no evidence of cellular toxicity or apoptosis in CDU-treated cells when incubated with 20 microM CDU for up to 48 h. These results, in light of the antiinflammatory and antihypertensive properties of these compounds that have been demonstrated already, suggest that the urea class of sEH inhibitors may be useful for therapy for diseases such as hypertension and atherosclerosis characterized by exuberant VSM cell proliferation and vascular inflammation.
ESTHER : Davis_2002_Proc.Natl.Acad.Sci.U.S.A_99_2222
PubMedSearch : Davis_2002_Proc.Natl.Acad.Sci.U.S.A_99_2222
PubMedID: 11842228

Title : Urea and amide-based inhibitors of the juvenile hormone epoxide hydrolase of the tobacco hornworm (Manduca sexta: Sphingidae) - Severson_2002_Insect.Biochem.Mol.Biol_32_1741
Author(s) : Severson TF , Goodrow MH , Morisseau C , Dowdy DL , Hammock BD
Ref : Insect Biochemistry & Molecular Biology , 32 :1741 , 2002
Abstract : A new class of inhibitors of juvenile hormone epoxide hydrolase (JHEH) of Manduca sexta and further in vitro characterization of the enzyme are reported. The compounds are based on urea and amide pharmacophores that were previously demonstrated as effective inhibitors of mammalian soluble and microsomal epoxide hydrolases. The best inhibitors against JHEH activity so far within this class are N-[(Z)-9-octadecenyl]-N'-propylurea and N-hexadecyl-N'-propylurea, which inhibited hydrolysis of a surrogate substrate (t-DPPO) with an IC(50) around 90 nM. The importance of substitution number and type was investigated and results indicated that N, N'-disubstitution with asymmetric alkyl groups was favored. Potencies of pharmacophores decreased as follows: amide>urea>carbamate>carbodiimide>thiourea and thiocarbamate for N, N'-disubstituted compounds with symmetric substituents, and urea>amide>carbamate for compounds with asymmetric N, N'-substituents. JHEH hydrolyzes t-DPPO with a K(m) of 65.6 microM and a V(max) of 59 nmol min(-1) mg(-1) and has a substantially lower K(m) of 3.6 microM and higher V(max) of 322 nmol min(-1) mg(-1) for JH III. Although none of these compounds were potent inhibitors of hydrolysis of JH III by JHEH, they are the first leads toward inhibitors of JHEH that are not potentially subject to metabolism through epoxide degradation.
ESTHER : Severson_2002_Insect.Biochem.Mol.Biol_32_1741
PubMedSearch : Severson_2002_Insect.Biochem.Mol.Biol_32_1741
PubMedID: 12429126

Title : Soluble epoxide hydrolase inhibition lowers arterial blood pressure in angiotensin II hypertension - Imig_2002_Hypertension_39_690
Author(s) : Imig JD , Zhao X , Capdevila JH , Morisseau C , Hammock BD
Ref : Hypertension , 39 :690 , 2002
Abstract : Epoxyeicosatrienoic acids (EETs) have antihypertensive properties and play a part in the maintenance of renal microvascular function. A novel approach to increase EET levels is to inhibit epoxide hydrolase enzymes that are responsible for conversion of biologically active EETs to dihydroxyeicosatrienoic acids (DHETs) that are void of effects on the preglomerular vasculature. We hypothesized that inhibition of soluble epoxide hydrolase (sEH) would lower blood pressure in angiotensin II (Ang II) hypertension. Rat renal cortical tissue was harvested and urine collected 2 weeks following implantation of an osmotic minipump containing Ang II (60 ng/min). Renal cortical sEH protein expression was significantly higher in Ang II hypertension compared with normotensive animals. Likewise, urinary 14,15-DHET levels were significantly increased in hypertensive compared with normotensive animals and averaged 8.1 +/- 1.3 and 2.7 +/- 1.1 ng/d; respectively. In additional experiments, the sEH inhibitor N-cyclohexyl-N-dodecyl urea (NCND; 3 mg/d) or vehicle (corn oil, 0.5 mL) was administered daily by intraperitoneal injection starting on day 10. Administration of NCND for 4 days lowered systolic blood pressure by 30 mm Hg in Ang II hypertensive animals, whereas the corn oil vehicle had no effect on blood pressure in normotensive or Ang II hypertensive animals. Measurement of blood pressure by indwelling arterial catheters in conscious animals with free movement in their cages confirmed that NCND had antihypertensive properties. Arterial blood pressure averaged 119 +/- 5 mm Hg in normotensive, 170 +/- 3 mm Hg in hypertensive and 149 +/- 10 mm Hg in NCND-treated, Ang II-infused animals. Administration of the potential metabolite of NCND, N-cyclohexylformamide to Ang II hypertensive rats did not lower the systolic blood pressure. These studies demonstrate that increased sEH expression in the Ang II hypertensive kidney leads to increased EET hydration. Moreover, sEH plays a role in the regulation of blood pressure, and inhibition of sEH during Ang II hypertension is antihypertensive.
ESTHER : Imig_2002_Hypertension_39_690
PubMedSearch : Imig_2002_Hypertension_39_690
PubMedID: 11882632

Title : Inhibition of microsomal epoxide hydrolases by ureas, amides, and amines - Morisseau_2001_Chem.Res.Toxicol_14_409
Author(s) : Morisseau C , Newman JW , Dowdy DL , Goodrow MH , Hammock BD
Ref : Chemical Research in Toxicology , 14 :409 , 2001
Abstract : The microsomal epoxide hydrolase (mEH) plays a significant role in the metabolism of xenobiotics such as polyaromatic toxicants. Additionally, polymorphism studies have underlined a potential role of this enzyme in relation to several diseases, such as emphysema, spontaneous abortion, and several forms of cancer. To provide new tools for studying the function of mEH, inhibition of this enzyme was investigated. Inhibition of recombinant rat and human mEH was achieved using primary ureas, amides, and amines. Several of these compounds are more potent than previously published inhibitors. Elaidamide, the most potent inhibitor that is obtained, has a K(i) of 70 nM for recombinant rat mEH. This compound interacts with the enzyme forming a noncovalent complex, and blocks substrate turnover through an apparent mix of competitive and noncompetitive inhibition kinetics. Furthermore, in insect cell cultures expressing rat mEH, elaidamide enhances the toxicity effects of epoxide-containing xenobiotics. These inhibitors could be valuable tools for investigating the physiological and toxicological roles of mEH.
ESTHER : Morisseau_2001_Chem.Res.Toxicol_14_409
PubMedSearch : Morisseau_2001_Chem.Res.Toxicol_14_409
PubMedID: 11304129

Title : Pathways of epoxyeicosatrienoic acid metabolism in endothelial cells. Implications for the vascular effects of soluble epoxide hydrolase inhibition - Fang_2001_J.Biol.Chem_276_14867
Author(s) : Fang X , Kaduce TL , Weintraub NL , Harmon S , Teesch LM , Morisseau C , Thompson DA , Hammock BD , Spector AA
Ref : Journal of Biological Chemistry , 276 :14867 , 2001
Abstract : Epoxyeicosatrienoic acids (EETs) are products of cytochrome P-450 epoxygenase that possess important vasodilating and anti-inflammatory properties. EETs are converted to the corresponding dihydroxyeicosatrienoic acid (DHET) by soluble epoxide hydrolase (sEH) in mammalian tissues, and inhibition of sEH has been proposed as a novel approach for the treatment of hypertension. We observed that sEH is present in porcine coronary endothelial cells (PCEC), and we found that low concentrations of N,N'-dicyclohexylurea (DCU), a selective sEH inhibitor, have profound effects on EET metabolism in PCEC cultures. Treatment with 3 microM DCU reduced cellular conversion of 14,15-EET to 14,15-DHET by 3-fold after 4 h of incubation, with a concomitant increase in the formation of the novel beta-oxidation products 10,11-epoxy-16:2 and 8,9-epoxy-14:1. DCU also markedly enhanced the incorporation of 14,15-EET and its metabolites into PCEC lipids. The most abundant product in DCU-treated cells was 16,17-epoxy-22:3, the elongation product of 14,15-EET. Another novel metabolite, 14,15-epoxy-20:2, was present in DCU-treated cells. DCU also caused a 4-fold increase in release of 14,15-EET when the cells were stimulated with a calcium ionophore. Furthermore, DCU decreased the conversion of [3H]11,12-EET to 11,12-DHET, increased 11,12-EET retention in PCEC lipids, and produced an accumulation of the partial beta-oxidation product 7,8-epoxy-16:2 in the medium. These findings suggest that in addition to being metabolized by sEH, EETs are substrates for beta-oxidation and chain elongation in endothelial cells and that there is considerable interaction among the three pathways. The modulation of EET metabolism by DCU provides novel insight into the mechanisms by which pharmacological or molecular inhibition of sEH effectively treats hypertension.
ESTHER : Fang_2001_J.Biol.Chem_276_14867
PubMedSearch : Fang_2001_J.Biol.Chem_276_14867
PubMedID: 11278979

Title : Evaluation of fish models of soluble epoxide hydrolase inhibition - Newman_2001_Environ.Health.Perspect_109_61
Author(s) : Newman JW , Denton DL , Morisseau C , Koger CS , Wheelock CE , Hinton DE , Hammock BD
Ref : Environmental Health Perspectives , 109 :61 , 2001
Abstract : Substituted ureas and carbamates are mechanistic inhibitors of the soluble epoxide hydrolase (sEH). We screened a set of chemicals containing these functionalities in larval fathead minnow (Pimphales promelas) and embryo/larval golden medaka (Oryzias latipes) models to evaluate the utility of these systems for investigating sEH inhibition in vivo. Both fathead minnow and medaka sEHs were functionally similar to the tested mammalian orthologs (murine and human) with respect to substrate hydrolysis and inhibitor susceptibility. Low lethality was observed in either larval or embryonic fish exposed to diuron [N-(3,4-dichlorophenyl), N'-dimethyl urea], desmethyl diuron [N-(3,4-dichlorophenyl), N'-methyl urea], or siduron [N-(1-methylcyclohexyl), N'-phenyl urea]. Dose-dependent inhibition of sEH was a sublethal effect of substituted urea exposure with the potency of siduron < desmethyl diuron = diuron, differing from the observed in vitro sEH inhibition potency of siduron > desmethyl diuron > diuron. Further, siduron exposure synergized the toxicity of trans-stilbene oxide in fathead minnows. Medaka embryos exposed to diuron, desmethyl diuron, or siduron displayed dose-dependent delays in hatch, and elevated concentrations of diuron and desmethyl diuron produced developmental toxicity. The dose-dependent toxicity and in vivo sEH inhibition correlated, suggesting a potential, albeit undefined, relationship between these factors. Additionally, the observed inversion of in vitro to in vivo potency suggests that these fish models may provide tools for investigating the in vivo stability of in vitro inhibitors while screening for untoward effects.
ESTHER : Newman_2001_Environ.Health.Perspect_109_61
PubMedSearch : Newman_2001_Environ.Health.Perspect_109_61
PubMedID: 11171526

Title : The role of methyl-linoleic acid epoxide and diol metabolites in the amplified toxicity of linoleic acid and polychlorinated biphenyls to vascular endothelial cells - Slim_2001_Toxicol.Appl.Pharmacol_171_184
Author(s) : Slim R , Hammock BD , Toborek M , Robertson LW , Newman JW , Morisseau C , Watkins BA , Saraswathi V , Hennig B
Ref : Toxicol Appl Pharmacol , 171 :184 , 2001
Abstract : Selected dietary lipids may increase the atherogenic effects of environmental chemicals, such as polychlorinated biphenyls (PCBs), by cross-amplifying mechanisms leading to dysfunction of the vascular endothelium. We have shown previously that the omega-6 parent fatty acid, linoleic acid, or 3,3',4,4'-tetrachlorobiphenyl (PCB 77), an aryl hydrocarbon (Ah) receptor agonist, independently can cause disruption of endothelial barrier function. Furthermore, cellular enrichment with linoleic acid can amplify PCB-induced endothelial cell dysfunction. We hypothesize that the amplified toxicity of linoleic acid and PCBs to endothelial cells could be mediated in part by cytotoxic epoxide metabolites of linoleic acid called leukotoxins (LTX) or their diol derivatives (LTXD). Exposure to LTXD resulted in a dose-dependent increase in albumin transfer across endothelial cell monolayers, whereas this disruption of endothelial barrier function was observed only at a high concentration of LTX. Pretreatment with the cytosolic epoxide hydrolase inhibitor 1-cyclohexyl-3-dodecyl urea partially protected against the observed LTX-induced endothelial dysfunction. Endothelial cell activation mediated by LTX and/or LTXD also enhanced nuclear translocation of the transcription factor NF-kappa B and gene expression of the inflammatory cytokine IL-6. Inhibiting cytosolic epoxide hydrolase decreased the LTX-mediated induction of both NF-kappa B and the IL-6 gene, whereas the antioxidant vitamin E did not block LTX-induced endothelial cell activation. Most importantly, inhibition of cytosolic epoxide hydrolase blocked both linoleic acid-induced cytotoxicity, as well as the additive toxicity of linoleic acid plus PCB 77 to endothelial cells. Interestingly, cellular uptake and accumulation of linoleic acid was markedly enhanced in the presence of PCB 77. These data suggest that cytotoxic epoxide metabolites of linoleic acid play a critical role in linoleic acid-induced endothelial cell dysfunction. Furthermore, the severe toxicity of PCBs in the presence of linoleic acid may be due in part to the generation of epoxide and diol metabolites. These findings have implications in understanding interactive mechanisms of how dietary fats can modulate dysfunction of the vascular endothelium mediated by certain environmental contaminants.
ESTHER : Slim_2001_Toxicol.Appl.Pharmacol_171_184
PubMedSearch : Slim_2001_Toxicol.Appl.Pharmacol_171_184
PubMedID: 11243918

Title : 3-D QSAR analysis of inhibition of murine soluble epoxide hydrolase (MsEH) by benzoylureas, arylureas, and their analogues - Nakagawa_2000_Bioorg.Med.Chem_8_2663
Author(s) : Nakagawa Y , Wheelock CE , Morisseau C , Goodrow MH , Hammock BG , Hammock BD
Ref : Bioorganic & Medicinal Chemistry , 8 :2663 , 2000
Abstract : Two hundred and seventy-one compounds including benzoylureas, arylureas and related compounds were assayed using recombinant murine soluble epoxide hydrolase (MsEH) produced from a baculovirus expression system. Among all the insect growth regulators assayed, 18 benzoylphenylurea congeners showed weak activity against MsEH. Newly synthesized cyclohexylphenylurea, 1-benzyl-3-phenylurea, and 1,3-dibenzylurea analogues were rather potent. The introduction of a methyl group at the para-position of the phenyl ring of cyclohexylphenylurea enhanced the activity 6-fold, though similar substituent effects were not seen for any of the benzoylphenylureas. The activities of these compounds, including several previously reported compounds, such as dicyclohexylurea, diphenylurea, and their related analogues (Morisseau et al., Proc. Natl. Acad. Sci., 1999, 96, 8849), were quantitatively analyzed using comparative molecular field analysis (CoMFA), a three-dimensional quantitative structure-activity relationship (3-D QSAR) method. Both steric and electrostatic factors contributing to variations in the activity were visualized using CoMFA. CoMFA results showed that one side of the cyclohexylurea moiety having a trans-amide conformation (A-ring moiety) is surrounded by large sterically unfavorable fields, while the other side of A-ring moiety and the other cyclohexyl group (B-ring moiety) is encompassed by sterically favored fields. Electrostatically negative fields were scattered around the entire molecule, and a positive field surrounds the carbon of the carbonyl group. Hydrophobic fields were visualized using Kellogg's hydropathic interaction (HINT) in conjunction with CoMFA. Hydrophobically favorable fields appeared beside the 4- and 4'-carbon atoms of the cyclohexyl groups, and hydrophobically unfavorable fields surrounded the urea bridge. The addition of the molecular hydrophobicity, log P [corrected], to CoMFA did not improve the correlation significantly. The ligand-binding interactions shown by X-ray crystallographic data were rationalized using the results of the CoMFA and HINT analyses, and the essential physicochemical parameters for the design of new MsEH inhibitors were disclosed.
ESTHER : Nakagawa_2000_Bioorg.Med.Chem_8_2663
PubMedSearch : Nakagawa_2000_Bioorg.Med.Chem_8_2663
PubMedID: 11092551

Title : Cress and potato soluble epoxide hydrolases: purification, biochemical characterization, and comparison to mammalian enzymes - Morisseau_2000_Arch.Biochem.Biophys_378_321
Author(s) : Morisseau C , Beetham JK , Pinot F , Debernard S , Newman JW , Hammock BD
Ref : Archives of Biochemistry & Biophysics , 378 :321 , 2000
Abstract : Affinity chromatographic methods were developed for the one-step purification to homogeneity of recombinant soluble epoxide hydrolases (sEHs) from cress and potato. The enzymes are monomeric, with masses of 36 and 39 kDa and pI values of 4.5 and 5.0, respectively. In spite of a large difference in sequence, the two plant enzymes have properties of inhibition and substrate selectivity which differ only slightly from mammalian sEHs. Whereas mammalian sEHs are highly selective for trans- versus cis-substituted stilbene oxide and 1,3-diphenylpropene oxide (DPPO), plant sEHs exhibit far greater selectivity for trans- versus cis-stilbene oxide, but little to no selectivity for DPPO isomers. The isolation of a covalently linked plant sEH-substrate complex indicated that the plant and mammalian sEHs have a similar mechanism of action. We hypothesize an in vivo role for plant sEH in cutin biosynthesis, based on relatively high plant sEH activity on epoxystearate to form a cutin precursor, 9,10-dihydroxystearate. Plant sEHs display a high thermal stability relative to mammalian sEHs. This stability and their high enantioselectivity for a single substrate suggest that their potential as biocatalysts for the preparation of enantiopure epoxides should be evaluated.
ESTHER : Morisseau_2000_Arch.Biochem.Biophys_378_321
PubMedSearch : Morisseau_2000_Arch.Biochem.Biophys_378_321
PubMedID: 10860549
Gene_locus related to this paper: arath-At2g26740

Title : Biochemical evidence for the involvement of tyrosine in epoxide activation during the catalytic cycle of epoxide hydrolase - Yamada_2000_J.Biol.Chem_275_23082
Author(s) : Yamada T , Morisseau C , Maxwell JE , Argiriadi MA , Christianson DW , Hammock BD
Ref : Journal of Biological Chemistry , 275 :23082 , 2000
Abstract : Epoxide hydrolases (EH) catalyze the hydrolysis of epoxides and arene oxides to their corresponding diols. The crystal structure of murine soluble EH suggests that Tyr(465) and Tyr(381) act as acid catalysts, activating the epoxide ring and facilitating the formation of a covalent intermediate between the epoxide and the enzyme. To explore the role of these two residues, mutant enzymes were produced and the mechanism of action was analyzed. Enzyme assays on a series of substrates confirm that both Tyr(465) and Tyr(381) are required for full catalytic activity. The kinetics of chalcone oxide hydrolysis show that mutation of Tyr(465) and Tyr(381) decreases the rate of binding and the formation of an intermediate, suggesting that both tyrosines polarize the epoxide moiety to facilitate ring opening. These two tyrosines are, however, not implicated in the hydrolysis of the covalent intermediate. Sequence comparisons showed that Tyr(465) is conserved in microsomal EHs. The substitution of analogous Tyr(374) with phenylalanine in the human microsomal EH dramatically decreases the rate of hydrolysis of cis-stilbene oxide. These results suggest that these tyrosines perform a significant mechanistic role in the substrate activation by EHs.
ESTHER : Yamada_2000_J.Biol.Chem_275_23082
PubMedSearch : Yamada_2000_J.Biol.Chem_275_23082
PubMedID: 10806198

Title : Soluble epoxide hydrolase regulates hydrolysis of vasoactive epoxyeicosatrienoic acids - Yu_2000_Circ.Res_87_992
Author(s) : Yu Z , Xu F , Huse LM , Morisseau C , Draper AJ , Newman JW , Parker C , Graham L , Engler MM , Hammock BD , Zeldin DC , Kroetz DL
Ref : Circulation Research , 87 :992 , 2000
Abstract : The cytochrome P450-derived epoxyeicosatrienoic acids (EETs) have potent effects on renal vascular reactivity and tubular sodium and water transport; however, the role of these eicosanoids in the pathogenesis of hypertension is controversial. The current study examined the hydrolysis of the EETs to the corresponding dihydroxyeicosatrienoic acids (DHETs) as a mechanism for regulation of EET activity and blood pressure. EET hydrolysis was increased 5- to 54-fold in renal cortical S9 fractions from the spontaneously hypertensive rat (SHR) relative to the normotensive Wistar-Kyoto (WKY) rat. This increase was most significant for the 14,15-EET regioisomer, and there was a clear preference for hydrolysis of 14, 15-EET over the 8,9- and 11,12-EETs. Increased EET hydrolysis was consistent with increased expression of soluble epoxide hydrolase (sEH) in the SHR renal microsomes and cytosol relative to the WKY samples. The urinary excretion of 14,15-DHET was 2.6-fold higher in the SHR than in the WKY rat, confirming increased EET hydrolysis in the SHR in vivo. Blood pressure was decreased 22+/-4 mm Hg (P:<0.01) 6 hours after treatment of SHRs with the selective sEH inhibitor N:, N:'-dicyclohexylurea; this treatment had no effect on blood pressure in the WKY rat. These studies identify sEH as a novel therapeutic target for control of blood pressure. The identification of a potent and selective inhibitor of EET hydrolysis will be invaluable in separating the vascular effects of the EET and DHET eicosanoids.
ESTHER : Yu_2000_Circ.Res_87_992
PubMedSearch : Yu_2000_Circ.Res_87_992
PubMedID: 11090543

Title : Binding of alkylurea inhibitors to epoxide hydrolase implicates active site tyrosines in substrate activation - Argiriadi_2000_J.Biol.Chem_275_15265
Author(s) : Argiriadi MA , Morisseau C , Goodrow MH , Dowdy DL , Hammock BD , Christianson DW
Ref : Journal of Biological Chemistry , 275 :15265 , 2000
Abstract : The structures of two alkylurea inhibitors complexed with murine soluble epoxide hydrolase have been determined by x-ray crystallographic methods. The alkyl substituents of each inhibitor make extensive hydrophobic contacts in the soluble epoxide hydrolase active site, and each urea carbonyl oxygen accepts hydrogen bonds from the phenolic hydroxyl groups of Tyr(381) and Tyr(465). These hydrogen bond interactions suggest that Tyr(381) and/or Tyr(465) are general acid catalysts that facilitate epoxide ring opening in the first step of the hydrolysis reaction; Tyr(465) is highly conserved among all epoxide hydrolases, and Tyr(381) is conserved among the soluble epoxide hydrolases. In one enzyme-inhibitor complex, the urea carbonyl oxygen additionally interacts with Gln(382). If a comparable interaction occurs in catalysis, then Gln(382) may provide electrostatic stabilization of partial negative charge on the epoxide oxygen. The carboxylate side chain of Asp(333) accepts a hydrogen bond from one of the urea NH groups in each enzyme-inhibitor complex. Because Asp(333) is the catalytic nucleophile, its interaction with the partial positive charge on the urea NH group mimics its approach toward the partial positive charge on the electrophilic carbon of an epoxide substrate. Accordingly, alkylurea inhibitors mimic features encountered in the reaction coordinate of epoxide ring opening, and a structure-based mechanism is proposed for leukotoxin epoxide hydrolysis.
ESTHER : Argiriadi_2000_J.Biol.Chem_275_15265
PubMedSearch : Argiriadi_2000_J.Biol.Chem_275_15265
PubMedID: 10747889

Title : Metabolism of monoepoxides of methyl linoleate: bioactivation and detoxification - Greene_2000_Arch.Biochem.Biophys_376_420
Author(s) : Greene JF , Williamson KC , Newman JW , Morisseau C , Hammock BD
Ref : Archives of Biochemistry & Biophysics , 376 :420 , 2000
Abstract : Leukotoxin (ltx) and isoleukotoxin (iltx) methyl esters, are metabolites of methyl linoleic acid, an essential fatty acid. They have been associated with acute respiratory distress syndrome. The observed toxicity of ltx and iltx is, in fact, due to the metabolism of the epoxides to their corresponding diols by soluble epoxide hydrolase (sEH). Herein, we demonstrate that ltx/iltx are toxic in a time-dependent manner to human sEH expressing cells with a LT(50) of 10.6 +/- 0.8 h and that ltx and iltx have K(M) of 6.15 +/- 1.0 and 5. 17 +/- 0.56 microM, respectively, and V(max) of 2.67 +/- 0.04 and 1. 86 +/- 0.06 micromol/min/mg, respectively, which can be inhibited by sEH inhibitors. We show that four major metabolites of ltx/iltx are formed in our system, including ltx/iltx free acid, ltxd/iltxd, free acid, and phosphotidylcholine and phosphotidylethanolamine containing the carboxylic acid forms of both ltx/iltx and ltxd/iltxd, but that the only metabolite associated with toxicity is the carboxylic acid form of ltxd/iltxd, suggesting the involvement of cellular esterases. We demonstrate that a serine esterase inhibitor provides some protection from the toxicity of epoxy fatty esters to sEH expressing cells as do intercellular free sulfhydryls, but that this protection is not due to glutathione conjugation. With these data, we have proposed an extension of the metabolic pathway for ltx/iltx in eukaryotic cells.
ESTHER : Greene_2000_Arch.Biochem.Biophys_376_420
PubMedSearch : Greene_2000_Arch.Biochem.Biophys_376_420
PubMedID: 10775430

Title : Detoxification of environmental mutagens and carcinogens: structure, mechanism, and evolution of liver epoxide hydrolase - Argiriadi_1999_Proc.Natl.Acad.Sci.U.S.A_96_10637
Author(s) : Argiriadi MA , Morisseau C , Hammock BD , Christianson DW
Ref : Proceedings of the National Academy of Sciences of the United States of America , 96 :10637 , 1999
Abstract : The crystal structure of recombinant murine liver cytosolic epoxide hydrolase (EC 3.3.2.3) has been determined at 2.8-A resolution. The binding of a nanomolar affinity inhibitor confirms the active site location in the C-terminal domain; this domain is similar to that of haloalkane dehalogenase and shares the alpha/beta hydrolase fold. A structure-based mechanism is proposed that illuminates the unique chemical strategy for the activation of endogenous and man-made epoxide substrates for hydrolysis and detoxification. Surprisingly, a vestigial active site is found in the N-terminal domain similar to that of another enzyme of halocarbon metabolism, haloacid dehalogenase. Although the vestigial active site does not participate in epoxide hydrolysis, the vestigial domain plays a critical structural role by stabilizing the dimer in a distinctive domain-swapped architecture. Given the genetic and structural relationships among these enzymes of xenobiotic metabolism, a structure-based evolutionary sequence is postulated.
ESTHER : Argiriadi_1999_Proc.Natl.Acad.Sci.U.S.A_96_10637
PubMedSearch : Argiriadi_1999_Proc.Natl.Acad.Sci.U.S.A_96_10637
PubMedID: 10485878
Gene_locus related to this paper: mouse-hyes

Title : Potent urea and carbamate inhibitors of soluble epoxide hydrolases - Morisseau_1999_Proc.Natl.Acad.Sci.U.S.A_96_8849
Author(s) : Morisseau C , Goodrow MH ,