%0 Journal Article %1 Lee2013FRETResidenceTimeSEH %A Lee, Kin Sing Stephen %A Morisseau, Christophe %A Yang, Jun %A Wang, Peng %A Hwang, Sung Hee %A Hammock, Bruce D. %D 2013 %J Analytical Biochemistry %K drug-kinetics fret k_off-measurements ligand-binding ligand-binding-kinetics residence-time residence-time-drug-design %N 2 %P 259 - 268 %R http://dx.doi.org/10.1016/j.ab.2012.11.015 %T Förster resonance energy transfer competitive displacement assay for human soluble epoxide hydrolase %U http://www.sciencedirect.com/science/article/pii/S0003269712005957 %V 434 %X 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 (Ki) for these inhibitors using catalytic assay is laborious. In addition, koff, 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 Förster resonance energy transfer (FRET)-based competitive displacement assay using intrinsic tryptophan fluorescence from sEH. In addition, the resulting assay allows us to measure the Ki values of very potent compounds to the picomolar level and to obtain relative koff values of the inhibitors. This assay provides additional data to evaluate the potency of sEH inhibitors.

@article{Lee2013FRETResidenceTimeSEH, 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 (Ki) for these inhibitors using catalytic assay is laborious. In addition, koff, 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 Förster resonance energy transfer (FRET)-based competitive displacement assay using intrinsic tryptophan fluorescence from sEH. In addition, the resulting assay allows us to measure the Ki values of very potent compounds to the picomolar level and to obtain relative koff values of the inhibitors. This assay provides additional data to evaluate the potency of sEH inhibitors. }, added-at = {2016-06-02T20:05:06.000+0200}, author = {Lee, Kin Sing Stephen and Morisseau, Christophe and Yang, Jun and Wang, Peng and Hwang, Sung Hee and Hammock, Bruce D.}, biburl = {https://www.bibsonomy.org/bibtex/28aa1a15ad491690756883509212d5538/salotz}, description = {Förster resonance energy transfer competitive displacement assay for human soluble epoxide hydrolase}, doi = {http://dx.doi.org/10.1016/j.ab.2012.11.015}, interhash = {59b3f6b09cba7d09bc3887b7db97257f}, intrahash = {8aa1a15ad491690756883509212d5538}, issn = {0003-2697}, journal = {Analytical Biochemistry }, keywords = {drug-kinetics fret k_off-measurements ligand-binding ligand-binding-kinetics residence-time residence-time-drug-design}, number = 2, pages = {259 - 268}, timestamp = {2016-06-02T20:05:06.000+0200}, title = {Förster resonance energy transfer competitive displacement assay for human soluble epoxide hydrolase }, url = {http://www.sciencedirect.com/science/article/pii/S0003269712005957}, volume = 434, year = 2013 }