ChEMBL ligand: CHEMBL436774 (Ar9281)
epoxide hydrolase 2/Bifunctional epoxide hydrolase 2 in Human [ChEMBL: CHEMBL2409] [GtoPdb: 2970] [UniProtKB: P34913] epoxide hydrolase 2/Bifunctional epoxide hydrolase 2 in Mouse [ChEMBL: CHEMBL4140] [GtoPdb: 2970] [UniProtKB: P34914] epoxide hydrolase 2/Bifunctional epoxide hydrolase 2 in Rat [ChEMBL: CHEMBL5669] [GtoPdb: 2970] [UniProtKB: P80299] There should be some charts here, you may need to enable JavaScript!
Epoxide hydrolase 1 in Human [ChEMBL: CHEMBL1968] [UniProtKB: P07099] There should be some charts here, you may need to enable JavaScript!

DB	Assay description	Assay Type	Standard value	Standard parameter	Original value	Original units	Original parameter	Reference
epoxide hydrolase 2/Bifunctional epoxide hydrolase 2 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL2409] [GtoPdb: 2970] [UniProtKB: P34913]
ChEMBL	Binding affinity to purified recombinant human sEH by FRET-displacement assay	B	7.71	pKi	19.5	nM	Ki	J Med Chem (2021) 64: 1856-1872 [PMID:33550801]
ChEMBL	FRET-displacement assay: FRET assays to determine Ki for the compounds of Table I were carried out as described previously (Lee et al. Analytical Biochemistry 434 (2013) 259-268). In order to prevent leaching of fluorescence impurities from the plastic tube and non-specific binding to sEH inhibitors, the inhibitor stock solution (10 mM, DMSO) was stored in glass vials. In addition, sEH was diluted to desired concentration (20 nM) with sodium phosphate buffer (PB) (100 mM sodium phosphate, pH 7.4, 0.01% gelatin) to avoid loss of protein from non-specific binding to the cuvette surface. All buffer used in this assay was filtered by sterilized filtration unit (Millipore Durapore PVDF Membrane, pore size: 0.22 um).Measurement in 96-Well PlatesAll the measurement for FRET-based displacement assay in 96-well plate format were done in TECAN Infinite M1000 Pro 96 well fluorescence plate reader.Pre-Treatment of 96-Well PlateIn order to prevent non-specific binding of sEH or inhibitor on the 96-well plate, the 96 well plates were pre-incubated with PB with 0.1% gelatin overnight at rt. The gelatin coats the plate and prevents non-specific binding of sEH and sEH inhibitors to the plate. The buffer was discarded and the plate was dried before use.Assay ProcedureThe sEH stock was diluted to the desired concentration (20 nM) by PB (100 mM sodium phosphate, 0.1% gelatin, pH 7.4). ACPU (one equivalent to sEH, 10 mM, Ethanol) was added to the sEH solution and was incubated for 2 h at rt. The sEH-ACPU mixture (20 nM, 100 mM sodium phosphate, 0.1% gelatin, pH 7.4, 150 uL) was added to each well.The baseline fluorescence (F0) (λexcitation at 280 nm, λemission at 450 nm) of the samples was measured after the z-position and gain were optimized automatically by the fluorometers. The z and gain value was noted and will be used for the later fluorescent measurement. Because DMSO has been known to quench fluorescence. 1% DMSO in PB was served as a control (FDMSO). The desired concentration of inhibitors which is the concentration that 100% of sEH was bound to inhibitor, was added at the first well and was further diluted by 2-fold across the rest of the wells. Based on our study, 12 datum points which correspond to 12 different concentrations of the inhibitor, provide significant data to calculate the accurate Ki for the inhibitors. The samples were incubated at 30° C. for 1.5 h. Then, the fluorescence (λexcitation at 280 nm, λemission at 450 nm) of the samples was measured using the z-position and gain values that previously obtained.	B	7.71	pKi	19.5	nM	Ki	US-10377744-B2. Potent soluble epdxide hydrolase inhibitors (2019)
ChEMBL	Assays for Epoxide Hydrolase Activity: Any of a number of standard assays for determining epoxide hydrolase activity can be used to determine inhibition of sEH. For example, suitable assays are described in Gill, et al., Anal Biochem 131:273-282 (1983); and Borhan, et al., Analytical Biochemistry 231:188-200 (1995)). Suitable in vitro assays are described in Zeldin et al., J Biol. Chem. 268:6402-6407 (1993). Suitable in vivo assays are described in Zeldin et al., Arch Biochem Biophys 330:87-96 (1996). Assays for epoxide hydrolase using both putative natural substrates and surrogate substrates have been reviewed (see, Hammock, et al. In: Methods in Enzymology, Volume III, Steroids and Isoprenoids, Part B, (Law, J. H. and H. C. Rilling, eds. 1985), Academic Press, Orlando, Fla., pp. 303-311 and Wixtrom et al., In: Biochemical Pharmacology and Toxicology, Vol. 1: Methodological Aspects of Drug Metabolizing Enzymes, (Zakim, D. and D. A. Vessey, eds. 1985), John Wiley & Sons, Inc., New York, pp. 1-93. Several spectral based assays exist based on the reactivity or tendency of the resulting diol product to hydrogen bond (see, e.g., Wixtrom, supra, and Hammock. Anal. Biochem. 174:291-299 (1985) and Dietze, et al. Anal. Biochem. 216:176-187 (1994)).The enzyme also can be detected based on the binding of specific ligands to the catalytic site which either immobilize the enzyme or label it with a probe such as dansyl, fluoracein, luciferase, green fluorescent protein or other reagent. The enzyme can be assayed by its hydration of EETs, its hydrolysis of an epoxide to give a colored product as described by Dietze et al., 1994, supra, or its hydrolysis of a radioactive surrogate substrate (Borhan et al., 1995, supra). The enzyme also can be detected based on the generation of fluorescent products following the hydrolysis of the epoxide. Numerous methods of epoxide hydrolase detection have been described (see, e.g., Wixtrom, supra).The assays are normally carried out with a recombinant enzyme following affinity purification. They can be carried out in crude tissue homogenates, cell culture or even in vivo, as known in the art and described in the references cited above.	B	7.71	pKi	19.5	nM	Ki	US-11123311-B2. Methods of treating mental disorders (2021)
ChEMBL	Inhibition of soluble epoxide hydrolase in HUVEC assessed inhibition of as conversion of 14, 15-EET to 14, 15-DHET	B	7.24	pIC50	57	nM	IC50	Bioorg Med Chem Lett (2011) 21: 983-988 [PMID:21211973]
ChEMBL	Inhibition of human sEH by fluorescent assay	B	7.82	pIC50	15	nM	IC50	J Med Chem (2007) 50: 3825-3840 [PMID:17616115]
ChEMBL	Inhibition of human recombinant sEH using PHOME as substrate measured after 10 mins by fluorescent based assay	B	7.86	pIC50	13.8	nM	IC50	Eur J Med Chem (2021) 223: 113678-113678 [PMID:34218083]
ChEMBL	Inhibition of recombinant human soluble epoxide hydrolase using CMNPC as substrate preincubated for 5 mins followed by substrate addition and measured at 30 secs interval for 10 mins by fluorescence assay	B	8.1	pIC50	8	nM	IC50	J Med Chem (2020) 63: 9237-9257 [PMID:32787085]
ChEMBL	Inhibition of sEH (unknown origin)	B	8.1	pIC50	8	nM	IC50	J Med Chem (2023) 66: 9201-9222 [PMID:37334504]
ChEMBL	Inhibition of human soluble epoxide hydrolase	B	8.1	pIC50	8	nM	IC50	Bioorg Med Chem Lett (2011) 21: 983-988 [PMID:21211973]
ChEMBL	Inhibition of recombinant human liver soluble epoxide hydrolase expressed in baculovirus infected Sf21 cells using NEPC as substrate by fluorescence based assay	B	8.1	pIC50	8	nM	IC50	Bioorg Med Chem (2019) 27: 115078-115078 [PMID:31488357]
ChEMBL	Inhibition of human recombinant sEH using CMNPC as substrate incubated for 5 mins by fluorescent based assay	B	8.1	pIC50	8	nM	IC50	J Med Chem (2021) 64: 5429-5446 [PMID:33945278]
ChEMBL	Inhibition of human recombinant sEH	B	8.1	pIC50	8	nM	IC50	J Med Chem (2022) 65: 13660-13680 [PMID:36222708]
GtoPdb	-	-	8.1	pIC50	8	nM	IC50	J Med Chem (2020) 63: 9237-9257 [PMID:32787085]
GtoPdb	-	-	8.15	pIC50	7	nM	IC50	Bioorg Med Chem Lett (2006) 16: 5212-6 [PMID:16870439]
ChEMBL	Inhibition of human soluble epoxide hydrolase using CMNPC as substrate after 5 mins by fluorescent assay	B	8.15	pIC50	7	nM	IC50	Bioorg Med Chem Lett (2012) 22: 601-605 [PMID:22079754]
ChEMBL	Inhibition of soluble epoxide hydroxylase by kinetic fluorescent assay	B	8.15	pIC50	7	nM	IC50	Bioorg Med Chem Lett (2006) 16: 5212-5216 [PMID:16870439]
epoxide hydrolase 2/Bifunctional epoxide hydrolase 2 in Mouse (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL4140] [GtoPdb: 2970] [UniProtKB: P34914]
ChEMBL	Inhibition of mouse recombinant soluble epoxide hydrolase by fluorescent assay	B	8.05	pIC50	9	nM	IC50	J Med Chem (2007) 50: 3825-3840 [PMID:17616115]
ChEMBL	Inhibition of mouse soluble epoxide hydrolase	B	8.52	pIC50	3	nM	IC50	Bioorg Med Chem Lett (2011) 21: 983-988 [PMID:21211973]
ChEMBL	Inhibition of murine soluble epoxide hydrolase expressed in baculovirus infected Sf21 cells using NEPC as substrate by fluorescence based assay	B	8.52	pIC50	3	nM	IC50	Bioorg Med Chem (2019) 27: 115078-115078 [PMID:31488357]
ChEMBL	Inhibition of mouse recombinant sEH using PHOME as substrate measured after 10 mins by fluorescent based assay	B	8.77	pIC50	1.7	nM	IC50	Eur J Med Chem (2021) 223: 113678-113678 [PMID:34218083]
GtoPdb	-	-	8.77	pIC50	1.7	nM	IC50	Eur J Med Chem (2021) 223: 113678 [PMID:34218083]
epoxide hydrolase 2/Bifunctional epoxide hydrolase 2 in Rat (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL5669] [GtoPdb: 2970] [UniProtKB: P80299]
ChEMBL	Inhibition of rat recombinant soluble epoxide hydrolase by fluorescent assay	B	8.22	pIC50	6	nM	IC50	J Med Chem (2007) 50: 3825-3840 [PMID:17616115]
Epoxide hydrolase 1 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL1968] [UniProtKB: P07099]
ChEMBL	Inhibition Assay: Inhibition assay of human soluble epoxide hydrolases.	B	7.84	pIC50	14.5	nM	IC50	US-8501783-B2. Conformationally restricted urea inhibitors of soluble epoxide hydrolase (2013)

ChEMBL data shown on this page come from version 36:

Zdrazil B, Felix E, Hunter F, Manners EJ, Blackshaw J, Corbett S, de Veij M, Ioannidis H, Lopez DM, Mosquera JF, Magarinos MP, Bosc N, Arcila R, Kizilören T, Gaulton A, Bento AP, Adasme MF, Monecke P, Landrum GA, Leach AR. (2024). The ChEMBL Database in 2023: a drug discovery platform spanning multiple bioactivity data types and time periods. Nucleic Acids Res., 52(D1). DOI: 10.1093/nar/gkad1004. [EPMCID:10767899] [PMID:37933841]
Davies M, Nowotka M, Papadatos G, Dedman N, Gaulton A, Atkinson F, Bellis L, Overington JP. (2015) 'ChEMBL web services: streamlining access to drug discovery data and utilities.' Nucleic Acids Res., 43(W1). DOI: 10.1093/nar/gkv352. [EPMCID:25883136]