Table 3.

Inhibition of human sEH by amide derivatives substituted with oxyoxalamide function

No.	R	R1	R2	R3	Human sEH IC50 a (nM)	Solubilityb (μM)
17d					>1000	40
18		H	H	CH3	280	NDc
19		H	H		408	ND
20		H	CH3	CH3	190	ND
21		CH3	CH3	CH3	204	ND
22		H	H		>1000	ND
23		H	CH3	CH3	69	156
24		CH3	CH3	CH3	7.9	625
25		H	CH3	CH3	35	78
26		CH3	CH3	CH3	4.4	156
2e					9.1	125
IK950e					14	625

^aTest compounds prepared in DMSO were reacted with human sEH (1 nM) for 10 min in 25 mM Bis–Tris/HCl buffer (202 μL; pH 7.0) at 30 °C. The fluorescent substrate (CMNPC; [S] = 5 μM) was then introduced to the incubation mixture. Inhibition potency against the human sEH was determined by measuring the appearance of the 6-methoxy-2-naphthaldehyde with an excitation wavelength of 330 nm and an emission wavelength of 465 nm for 10 min on a fluorometer. Results are averages of three separate measurements. See the Supplementary data for the detailed procedures.

^bWater solubility was determined by adding a variety of concentrations of a test compound prepared in DMSO to 0.1 M sodium phosphate buffer (pH 7.4) in a final ratio of 5:95 (v/v). The turbidity of the water solution was measured at 650 nm to determine solubility in water. Results are the average of triplicate determinations.

^cNot-determined because the inhibition results were not potent enough compared to that of other derivatives.

^dAmide inhibitor with no substitution by oxyoxalamide function.

^ePreviously reported potent and soluble inhibitors.^14,23 IK950 was used as a control compound for the measurement of water solubility in this study.