document: an imposing body of 
   studies , spanning 
   more than three decades , has cogently elucidated the involvement of 
   epoxyeicosatrienoic acids ( eets ) in a wide array of critical physiological 
   functions , inter alia , blood pressure regulation , nociception , adipogenesis , anti - inflammatory activity , organ regeneration , insulin 
   potentiation , podocyte integrity , and cellular responses to bacterial infection . 
   the most prominent regioisomer , 14,15-epoxyeicosa-5(z),8(z),11(z)-trienoic acid ( 14,15-eet ) , along with other 
   members of this autacoid family , arise via metabolism of arachidonic 
   acid by cytochromes p450 , especially members of the 2c and 2j families . 
   the ratio of eet isomers and their stereocomposition are cyp p450 
   isoform - dependent and , thus , species- 
   and tissue - specific . 
   numerous chemical 
   and metabolic factors conspire to complicate 
   investigations into the roles of 14,15-eet and restrain its potential 
   therapeutic utility ( figure 1 ) . 
   its susceptibility toward aerial oxidation or 
   auto - oxidation , i.e. , a nonenzymatic , free radical process involving 
   the 1,4-dienyl substructures characteristic of polyunsaturated fatty 
   acids , necessitates handling and/or storage under strict conditions 
   that minimize exposure to oxygen and trace transition metals . 
   a precis of inactivating enzymatic processes 
   includes esterification , metabolism by 
   other pathways of the arachidonate cascade , conjugation , -oxidation , chain elongation , and hydrolysis of the epoxide . in biological 
   milieu , 
   hydration of the epoxide by soluble epoxide hydrolase ( seh ) 
   is a major determinant in maintaining the steady state levels of 14,15-eet , whose half - life has been estimated at no more 
   than seconds to minutes . 
   the pioneering 
   studies of hammock and others confirmed inhibition of seh can elevate eet 
   levels both in vitro and in vivo , thus offering an indirect means 
   for pharmacologic intervention in eet - mediated processes . 
   however , 
   this strategy might prove less efficacious than an agonist replacement 
   therapy whenever endogenous eet biosynthesis is compromised , for instance , 
   as a consequence of disease , inflammation , radiotherapy , aging , and/or 
   exposure to xenobiotics and drugs that inhibit cytochromes p450 . 
   to address 
   some of the stability limitations of natural eets , our 
   laboratories previously prepared several iterations of eet surrogates 
   with improved stability . 
   advanced versions were 
   evaluated for relaxation of precontracted bovine coronary artery rings 
   and for in vitro inhibition of recombinant human seh . 
   depending upon the bioisostere and its position along the 
   carbon chain , varying levels of vascular relaxation and/or seh inhibition 
   were observed . 
   generally , oxamides and n - ipr - amides displayed useful 14,15-eet agonist activities 
   but were modest to poor seh inhibitors . 
   the in vitro 
   success of this generation of analogues prompted us to consider further 
   structural iterations . 
   the surrogates 
   described herein modify the free carboxylic acid 
   of the previous generation of 14,15-eet mimics while , for the most 
   part , retaining some key structural features of the pharmacophore 
   identified earlier ( i.e. , cis--olefin and an epoxide bioisostere ) . 
   they were evaluated as described 
   before in precontracted bovine coronary artery rings for ( i ) % vasorelaxation 
   at 10 m relative to a 14,15-eet control , ( ii ) ec50 for vasorelaxation , and ( iii ) ic50 for seh inhibition 
   ( table 1 ) . 
   interestingly , 
   the simple expedient of conjugating the carboxylate with a short poly(ethylene 
   oxide ) ( peg ) unit improved both % vasorelaxation and the ec50 somewhat compared with the parent free acids regardless the type of epoxide bioisostere , viz . , n - ipr - amide 1 , urea 2 , and oxamide 3 ; on the other hand , the soluble 
   epoxide hydrolase ( seh ) inhibitory activity of urea 2 was significantly compromised versus the parent free acid ( ec50 7.5 m , ic50 46 nm ) . 
   linkage of the carboxylate 
   to the nitrogen of glycine ( 4 and 5 ) or 
   aspartic acid ( 6 ) resulted in a modest boost to the vasoactivities 
   while the seh ic50 of 6 dropped by an order 
   of magnitude compared to the parent free acid . 
   conversion to the n - phenyl and n - methylsulfonimides 7 and 8 , respectively , left the biological activities 
   virtually unchanged in all categories while the simplified phenylsulfonamide 9 led to an improvement in the ec50 by a factor 
   of 2 and smaller improvements in the other parameters . at 10 m 
   , 14,15-eet induces 
   85% of maximum vasorelaxation and its ed50 is 2.2 m . 
   for recombinant human seh , 
   given the generally lackluster behavior 
   of the esters and amides , 
   our focus changed to replacement of the carboxylate with a variety 
   of heterocyclic bioisosteres identified from literature sources . 
   for phosphonate 10 and sulfonate 11 , the increase in polarity did not improve potency as seen 
   with the peg esters . 
   interestingly , sulfonate 11 retained 
   its ability to suppress seh , in contrast to phosphonate 10 and peg ester 2 . 
   vasorelaxation by s - alkylthiocatechol 12 was poor , but its ic50 for seh was pushed down into the single digits . 
   the sulfone variant 13 regained some vasopotency , 
   probably due to an increase in the acidity of the phenol but not sufficiently 
   to be viable . 
   replacement of the phenol moiety in 12 with 
   a tetrazole ring produced 14 and was encouraging , but this trend was not continued in triazole 15 . 
   sequential oxidation of the sulfur to sulfoxamide 16 and sulfonamide 17 , as seen for 12  13 , incrementally improved the ec50 . 
   the ed50 jumped in the tetrazole bioisostere series 1821 , achieving a submicromolar value 
   for urea 19 ( 12-fold better than 14,15-eet ) , while the 
   % vasorelaxation versus 14,15-eet peaked . 
   factoring in a very respectable 
   low nanomolar ic50 , 19 is the best dual activity 
   analogue in the study and a leading candidate for further development . 
   notably , all three parameters began to erode , albeit minimally , in 
   the one - carbon homologue 20 and further in oxamide 21 . 
   the performance of another five - membered heterocycle , oxadiazol-5-one 22 , while acceptable , 
   was not comparable to urea 19 . 
   the closely related oxathiadiazole-2-oxides23 and 24 , on the 
   other hand , demonstrated good vasorelaxant activities ; seh inhibition 
   potency was consistent with our previous observations that ureas  n - ipr - amides . 
   the isomeric thioxo-1,2,4-oxadiazol25 registered a 6-fold improvement 
   in ed50 over 14,15-eet yet was a comparatively poor seh 
   inhibitor and , thus , is a good option whenever an eet agonist with 
   minimal impact on seh function is required . 
   the corresponding urea 
   thioxo - oxadiazol,26 , could 
   not sustain the vasopotency ; its ed50 sagged by almost 
   an order of magnitude versus 25 while at the same time 
   its seh ic50 improved by more than an order of magnitude . 
   a final variant on this heterocyclic theme , 5-thioxothiadiazol27 , fell short of expectations 
   on all levels and further study of this particular heterocycle was 
   terminated . despite its success in the glitazone series of antidiabetic 
   drugs , the 2,4-thiazolidinedione modification 
   proved disappointing when incorporated into 28 . 
   given 
   the larger size of the n-(4-hydroxy-2-benzothiazolyl)acetamide 
   bioisostere , the carbon chain utilized 
   in most of the analogues ( figure 1 ) was trimmed 
   by four carbons before attachment via the phenolic oxygen to generate 29 . 
   following inspection of molecular models , the carbon 
   chain was shortened even further and the olefin was deleted , leading 
   to n - ipr - amide 30 , 
   urea 31 , and oxamide 32 , whose total chain 
   length more closely resembled that of 14,15-eet . as hoped , the ec50 values increased relative to 29 , although the 
   % vasorelaxation was variable . 
   contrary to expectations , 
   there was no evident correlation between 
   the pka of the eet analogues and the ed50 for vasorelaxation ( figure 2 ) , suggesting 
   factors other than ionic interactions are involved in binding the 
   carboxylate at the putative eet binding site . plot of calculated pka vs ed50 ( m ) of analogues 
   in table 1 . 
   the protonated 
   form of analogues 6 , 11 , and 12 was used for the calculation . 
   the syntheses of tetrazole 19 and 
   thioxo-1,2,4-oxadiazol 25 are summarized in scheme 1 and illustrate the approach used to prepare the 
   other analogues . 
   semihydrogenation of the known acetylene 34 using p-2 nickel and h2 led to cis - olefin 35 that was subjected 
   to azidation using diphenylphosphoryl azide ( dppa ) under mitsunobu 
   conditions . 
   staudinger reduction of the product , azide 36 , led to primary amine 37 that was reacted without purification 
   with n - pentyl isocyanate to furnish urea 38 . 
   an uneventful series of functional group interchanges proceeding 
   through alcohol 39 , bromide 40 , and ending 
   with nitrile 41 set the stage for the zinc bromide mediated 
   annulation with sodium azide that delivered 
   tetrazole 19 . 
   reagents and conditions : ( a ) 
   p2-ni/(h2nch2)2 , h2 ( 1 
   atm ) , etoh , rt , 1 h ( 96% ) ; ( b ) diad / ph3p / ph2p(o)n3 , thf , 20 to 23 c , 4 h ( 72% ) ; ( c ) 
   ph3p , h2o / thf , rt , 12 h ( 76% ) ; ( d ) c5h11nco , thf , rt , 3 h ( 76% ) ; ( e ) nbu4nf , thf , rt , 12 h ( 8289% ) ; ( f ) cbr4/ph3p , ch2cl2 , rt , 2 h ( 8384% ) ; 
   ( g ) kcn , dmso , rt , 12 h ( 7881% ) ; ( h ) nan3/znbr2 , iproh / h2o ( 1:3 ) , 110 c , 
   18 h ( 76% ) ; ( i ) ho(co)2nh(ch2)3ch3 , edci , dmf , rt , 12 h ( 73% ) ; ( j ) h2noh 
   / na2co3 , meoh / h2o ( 4:1 ) , 60 c , 18 h ( 62% ) ; 
   ( k ) im2c(s ) , thf , rt , 45 min ( 63% ) . 
   following the same sequence of transformations as described above , 42 was converted into nitrile 45 . 
   condensation 
   of hydroxyimine 46 , obtained by addition of hydroxylamine to 45 , with 1,1-(thiocarbonyl)diimidazole 
   yielded thioxo-1,2,4-oxadiazol 25 as a crystalline solid . 
   final compounds were 
   judged 95% pure by hplc using a zorbax eclipse c18 column 
   ( 250 mm  4.6 mm ; agilent ) connected to an agilent 1200 api / lc - ms 
   using acetonitrile / water combinations as eluent unless otherwise noted . 
   nuclear magnetic resonance ( nmr ) spectra were recorded on varian 300 , 
   400 , or 500 spectrometers at operating frequencies of 300/400/500 
   mhz ( h ) or 75/100/125 mhz ( c ) in cdcl3 with tms as internal standard , unless otherwise stated . 
   h nmr data are reported as follows : chemical shift ( ppm ) , 
   multiplicity ( s = singlet , br s = broad singlet , d = doublet , t = 
   triplet , q = quartet , app q = apparent quartet , qn = quintet , app 
   qn = apparent quintet , m = multiplet ) , and coupling constant ( hz ) . 
   high resolution mass spectra ( hrms ) were obtained at ut - arlington 
   using a shimadzu it - tof mass spectrometer or at the medical college 
   of wisconsin by prof . 
   infrared ( ir ) spectra were 
   obtained using a perkinelmer spectrum 1000 fourier transform spectrometer . 
   melting points were measured using an optimelt from stanford research 
   systems and are uncorrected . 
   analytical thin layer chromatography 
   ( tlc ) used emd chemicals tlc silica gel 60 f254 plates ( 0.0400.063 
   mm ) with visualization by uv light and/or kmno4 or phosphomolybdic 
   acid ( pma ) solution followed by heating . 
   all oxygen and/or moisture 
   sensitive reactions were performed under an argon atmosphere using 
   oven - dried glassware and anhydrous solvents . 
   extracts were dried over 
   anhydrous na2so4 and filtered prior to removal 
   of all volatiles under reduced pressure . 
   chromatographic purifications 
   utilized preparative tlc or flash chromatography using prepacked sio2 columns on a combiflash rf200 chromatograph ( teledyne 
   isco ) . unless otherwise noted , yields refer to isolated , purified 
   material with spectral data consistent with assigned structures or , 
   if known , were in agreement with published data . 
   reagents were purchased 
   at the highest commercial quality available and used without further 
   purification , unless otherwise noted . 
   anhydrous solvents were dried 
   using a glass contours solvent system by passage through columns of 
   activated packing material under argon immediately prior to use . a mixture of 1-(12-cyanododec-5(z)-enyl)-3-n - pentylurea ( 41 ) . 
   ( 500 mg , 1.55 mmol ) , sodium azide ( 100 mg , 1.55 mmol ) , and zinc 
   bromide ( 335 mg , 1.48 mmol ) was heated at 110 c in 2-propanol / h2o ( 1:3 , 8 ml ) while stirring vigorously in a sealed tube . after 18 h , 
   the mixture was cooled to room temperature 
   and the ph was adjusted to 1 using aq hcl ( 3 n , 4 ml ) . 
   ethyl acetate 
   ( 10 ml ) was added , and the stirring was continued until no solid was 
   present . 
   the organic layer was isolated and the aqueous layer extracted 
   with etoac ( 2  25 ml ) . 
   the combined organic fractions were washed 
   with water ( 3  25 ml ) , dried , and concentrated in vacuo . 
   the 
   residue was purified by silica gel column chromatography to give the 
   tetrazole 19 ( 431 mg , 76% ) as a colorless solid , mp 205.6205.8 
   c . 
   h nmr ( cd3od , 
   300 mhz )  5.405.30 ( m , 2h ) , 3.063.11 ( m , 4h ) , 
   2.93 ( t , j = 8.0 hz , 2h ) , 1.982.10 ( m , 4h ) , 
   1.701.82 ( m , 2h ) , 1.241.50 ( m , 16h ) , 0.90 ( t , j = 7.6 hz , 3h ) . 
   c nmr ( cd3od , 75 
   mhz )  160.16 , 156.81 , 129.77 , 129.47 , 39.81 , 39.68 , 29.88 , 
   29.80 , 29.35 , 28.99 , 28.69 , 28.55 , 27.48 , 26.85 , 26.81 , 26.68 , 22.96 , 
   22.31 , 13.22 . 
   hrms calcd for c19h37n6o [ m + 1 ] 365.3029 , found 365.3030 . a mixture of n-(13-amino-13-(hydroxyimino)tridec-5(z)-enyl)-n - n - butyloxalamide ( 46 ) ( 100 mg , 0.27 mmol ) and 1,1-(thiocarbonyl)diimidazole ( 57 
   mg , 0.32 mmol ) in dry thf ( 5 ml ) was stirred at room temperature for 
   45 min . 
   the mixture was diluted with water ( 20 ml ) and extracted with 
   ethyl acetate ( 3  10 ml ) . 
   the combined organic extracts were 
   washed with water and dried , and the solvent was evaporated in vacuo . 
   the residue was dissolved in acetonitrile ( 5 ml ) to which was then 
   added dbu ( 61 mg , 0.40 mmol ) . after stirring at room temperature for 
   1 h , the mixture was diluted with water ( 10 ml ) , adjusted ph  
   4 with 1 n hcl , and extracted with ethyl acetate ( 3  10 ml ) 
   to give thioxo-1,2,4-oxadiazol 25 ( 71 mg , 63% ) as a white 
   solid , mp 110.6110.8 c . 
   h nmr ( 400 mhz )  8.90 ( br s , nh , 1h ) , 7.52 ( br s , nh , 
   2h ) , 5.285.40 ( m , 2h ) , 3.203.40 ( m , 4h ) , 2.59 ( t , j = 7.5 hz , 2h ) , 1.982.10 ( m , 4h ) , 1.211.70 
   ( m , 16h ) , 0.92 ( t , j = 7.3 hz , 3h ) . 
   c nmr ( 100 mhz )  160.12 , 160.08 , 153.31 , 130.62 , 129.46 , 39.93 , 
   39.85 , 31.35 , 29.33 , 28.94 , 28.89 , 28.68 , 27.02 , 26.84 , 26.69 , 23.96 , 
   20.23 , 13.90 . 
   nabh4 ( 82 
   mg , 2.28 mmol ) was added in portions with vigorous stirring to a room 
   temperature solution of ni(oac)24h2o ( 567 
   mg , 2.28 mmol ) in absolute ethanol ( 20 ml ) under a hydrogen atmosphere 
   ( 1 atm ) . after 15 min , 
   freshly distilled ethylenediamine ( 0.30 ml , 
   4.56 mmol ) was added to the black suspension , followed after a further 
   15 min by a solution of 12-(tert - butyldiphenylsilyloxy)dodec-5-yn-1-ol ( 34 ) ( 4.0 g , 9.16 mmol ) in absolute 
   etoh ( 10 ml ) . 
   after 1 h , the reaction mixture was diluted with et2o ( 20 ml ) and passed through a small bed of silica gel . 
   the 
   combined ethereal filtrates were concentrated under reduced pressure 
   to afford alcohol 35 ( 3.85 g , 96% ) as a colorless oil 
   sufficiently pure to be used directly in the next step . 
   h 
   nmr ( 300 mhz )  7.647.68 ( m , 4h ) , 7.347.42 ( m , 
   6h ) , 5.425.28 ( m , 2h ) , 3.63 ( t , j = 6.4 hz , 
   4h ) , 2.081.96 ( m , 4h ) , 1.501.60 ( m , 4h ) , 1.401.24 
   ( m , 10h ) , 1.04 ( s , 9h ) . 
   c nmr ( 100 mhz )  135.81 , 
   134.40 , 130.61 , 129.71 , 129.60 , 127.80 , 64.21 , 63.14 , 32.78 , 32.60 , 
   29.98 , 29.27 , 27.42 , 27.14 , 27.10 , 26.08 , 25.92 , 19.48 . 
   diisopropyl azodicarboxylate 
   ( diad ; 1.46 ml , 7.35 mmol ) was added dropwise to a 20 c 
   solution of pph3 ( 2.10 g , 8.0 mmol ) in dry thf ( 45 ml ) 
   under an argon atmosphere . after 10 min , a solution of 12-(tert - butyldiphenylsilyloxy)dodec-5(z)-en-1-ol 
   ( 35 ) ( 3.20 g , 7.35 mmol ) in dry thf ( 10 ml ) 
   after 30 min , the mixture was warmed to 0 c and diphenylphosphoryl 
   azide ( 1.58 ml , 7.35 mmol ) was added dropwise . after stirring 4 h 
   at rt , 
   the reaction mixture was quenched with water ( 150 ml ) and extracted 
   with etoac ( 2  100 ml ) . 
   the combined organic extracts were washed 
   with brine ( 100 ml ) , dried ( na2so4 ) , and concentrated 
   under reduced pressure . 
   the residue was purified by sio2 column chromatography eluting with 4% etoac / hexane to afford azide 36 ( 2.45 g , 72% ) . 
   h nmr ( 400 mhz )  7.647.68 
   ( m , 4h ) , 7.347.42 ( m , 6h ) , 5.285.42 ( m , 2h ) , 3.70 
   ( t , j = 5.8 hz , 2h ) , 3.27 ( t , j = 
   6.3 hz , 2h ) , 1.962.10 ( m , 4h ) , 1.241.64 ( m , 12h ) , 
   1.04 ( s , 9h ) . 
   c nmr ( 100 mhz )  135.84 , 134.41 , 
   130.93 , 129.75 , 129.12 , 127.83 , 64.22 , 51.62 , 32.81 , 29.93 , 29.30 , 
   28.68 , 27.46 , 27.14 , 27.02 , 26.90 , 25.96 , 19.49 . 
   ir ( neat ) 2930 , 2783 , 
   2331 , 2097 , 1106 cm . 
   triphenylphosphine ( 1.18 
   g , 4.50 mmol ) was added to a stirring solution of 1-tert - butyldiphenylsilyloxy-12-azidododec-7(z)-ene ( 36 ) ( 1.90 g , 4.10 mmol ) in thf ( 12 ml ) containing 10 drops 
   of deionized water . 
   after 12 h , the reaction mixture was diluted with 
   ch2cl2 ( 10 ml ) , dried , and concentrated in vacuo 
   to give amine 37 ( 1.36 g , 76% ) as a viscous , colorless 
   oil that was used directly in the next reaction without further purification . 
   h nmr ( 400 mhz )  7.627.68 
   ( m , 4h ) , 7.327.40 ( m , 6h ) , 5.305.40 ( m , 2h ) , 3.63 
   ( t , j = 5.2 hz , 2h ) , 2.62 ( t , j = 
   4.8 hz , 2h ) , 1.922.06 ( m , 4h ) , 1.401.58 ( m , 4h ) , 1.201.40 
   ( m , 8h ) , 1.03 ( s , 9h ) . 
   c nmr ( 100 mhz )  135.79 , 
   134.37 , 130.42 , 129.70 , 127.78 , 64.19 , 42.28 , 33.44 , 32.77 , 29.93 , 
   29.28 , 27.40 , 27.21 , 27.10 , 25.92 , 19.44 . 
   a solution of 1-tert - butyldiphenylsilyloxy-12-aminododec-7(z)-ene ( 37 ) ( 1.32 g , 3.0 mmol ) in thf ( 5 ml ) 
   was added dropwise to a stirring solution of n - pentyl 
   isocyanate ( 0.386 ml , 3.0 mmol ) in thf ( 10 ml ) . 
   after 3 h stirring 
   at room temperature , all volatiles were removed under reduced pressure 
   and the residue was purified by sio2 column chromatography 
   eluting with 20% etoac / hexane to afford urea 38 ( 1.26 
   g , 76% ) as a viscous oil . 
   h nmr ( 300 mhz )  7.607.70 
   ( m , 4h ) , 7.357.42 ( m , 6h ) , 5.285.42 ( m , 2h ) , 5.16 
   ( br s , nh , 2h ) , 3.65 ( t , j = 6.5 hz , 2h ) , 
   3.083.20 ( m , 4h ) , 1.962.08 ( m , 4h ) , 1.221.60 
   ( m , 18h ) , 1.02 ( s , 9h ) , 0.89 ( t , j = 7.3 hz , 3h ) . 
   c nmr ( 100 mhz )  159.23 , 135.80 , 134.24 , 130.52 , 129.74 , 
   129.49 , 127.82 , 64.22 , 40.62 , 40.54 , 32.80 , 30.33 , 29.95 , 29.37 , 29.32 , 
   27.46 , 27.34 , 27.18 , 27.11 , 25.97 , 22.71 , 19.46 , 14.29 . 
   hrms calcd 
   for c34h55n2o2si [ m + 
   1 ] 551.4033 , found 551.4032 . a mixture of 1-(12-(tert - butyldiphenylsilyloxy)dodec-5(z)-enyl)-3-n - pentylurea ( 38 ) ( 1.12 g , 2.0 mmol ) and tetra - n - butylammonium fluoride 
   ( 2.20 ml of 1 m soln in thf , 2.2 
   mmol ) in dry thf ( 10 ml ) was stirred at room temperature under an 
   argon atmosphere for 12 h and then evaporated to dryness in vacuo . 
   the residue was dissolved in etoac ( 50 ml ) and washed with water ( 30 
   ml ) , brine ( 30 ml ) , dried , and evaporated in vacuo . 
   purification of 
   the residue via sio2 column chromatography gave alcohol 39 ( 0.56 g , 89% ) as a colorless solid , mp 63.763.8 
   c . 
   h nmr ( 300 mhz )  5.255.42 ( m , 2h ) , 
   4.48 ( br s , nh , 2h ) , 3.64 ( d , j = 6.5 hz , 
   2h ) , 3.083.20 ( m , 4h ) , 1.962.14 ( m , 4h ) , 1.221.60 
   ( m , 18h ) , 0.88 ( t , j = 7.0 hz , 3h ) . 
   c nmr ( 125 mhz )  159.26 , 130.23 , 129.62 , 63.72 , 40.33 , 40.29 , 
   32.92 , 30.30 , 30.26 , 29.74 , 29.35 , 29.13 , 27.26 , 27.20 , 27.13 , 25.82 , 
   22.69 , 14.27 . 
   cbr4 ( 0.55 g , 1.66 
   mmol ) and pph3 ( 0.43 g , 1.66 mmol ) were added to a 0 c 
   solution of 1-(12-hydroxydodec-5(z)-enyl)-3-n - pentylurea ( 39 ) ( 0.43 g , 1.38 mmol ) in ch2cl2 ( 20 ml ) . 
   after 2 h at room temperature , the 
   reaction mixture was concentrated in vacuo and the residue was purified 
   via sio2 column chromatography to give 1-(12-bromododec-5(z)-enyl)-3-n - pentylurea ( 40 ) ( 0.43 g , 83% ) as a solid , mp 46.746.8 c . 
   h 
   nmr ( 300 mhz )  5.225.42 ( m , 2h ) , 4.40 ( br s , 2h ) , 3.42 
   ( t , j = 9.3 hz , 2h ) , 3.103.20 ( m , 4h ) , 1.982.10 
   ( m , 4h ) , 1.801.90 ( m , 2h ) , 1.251.55 ( m , 16h ) , 0.92 
   ( t , j = 7.2 hz , 3h ) . c nmr ( 100 mhz ) 
    159.51 , 130.14 , 129.69 , 40.48 , 40.39 , 34.20 , 32.96 , 30.34 , 
   29.67 , 29.36 , 28.58 , 28.25 , 27.31 , 27.27 , 27.17 , 22.68 , 14.26 . 
   hrms 
   calcd for c18h36brn2o [ m + 1 ] 375.2011 , found 375.2014 . a mixture of potassium cyanide 
   ( 0.23 g , 3.54 mmol ) and 1-(12-bromododec-5(z)-enyl)-3-n - pentylurea ( 40 ) ( 0.90 g , 2.40 mmol ) was stirred 
   in dmso ( 5 ml ) at room temperature . 
   after 12 h , the reaction mixture 
   was diluted with water ( 20 ml ) and extracted with ethyl acetate ( 2 
    50 ml ) . 
   the combined organic extracts were washed with water 
   ( 2  25 ml ) , brine ( 25 ml ) , dried ( na2so4 ) , and passed through a silica gel column to give nitrile 41 ( 0.62 g , 81% ) as a colorless solid , mp 5657 c . 
   h nmr ( 300 mhz )  5.295.40 ( m , 2h ) , 4.27 ( br 
   s , nh , 2h ) , 3.103.20 ( m , 4h ) , 2.34 ( t , j = 7.0 hz , 2h ) , 1.982.08 ( m , 4h ) 1.241.70 ( m , 18h ) , 
   0.89 ( t , j = 7.0 hz , 3h ) . 
   c nmr ( 125 
   mhz )  159.41 , 129.94 , 129.86 , 120.14 , 40.45 , 40.35 , 30.30 , 
   29.50 , 29.33 , 28.70 , 28.51 , 27.26 , 27.16 , 25.47 , 22.66 , 17.28 , 14.24 . 
   ir ( neat ) 2930 , 2281 , 2184 , 2042 , 1936 , 1613 , 1197 , 1042 cm . 
   hrms calcd for c19h36n3o [ m + 
   1 ] 322.2858 , found 322.2867 . a mixture of 
   2-(n - butylamino)-2-oxoacetic acid ( 0.40 g , 2.70 mmol ) , 1-tert - butyldiphenylsilyloxy-12-aminododec-7(z)-ene ( 37 ) ( 1.20 g , 2.70 mmol ) , 1-hydroxybenzotriazole 
   ( hobt ; 0.44 g , 3.30 mmol ) , and [ 1-(3-(dimethylamino)propyl)-3-ethylcarbodiimide 
   hydrochloride ] ( edci : 0.63 g , 3.30 mmol ) in dry dmf ( 5 ml ) was stirred 
   at room temperature overnight . 
   the reaction mixture was quenched with 
   water ( 30 ml ) and extracted into ethyl acetate ( 3  20 ml ) . 
   the 
   combined organic extracts were washed with water ( 2  10 ml ) 
   and brine ( 10 ml ) , dried , and concentrated in vacuo . 
   the residue was 
   purified by sio2 column chromatography to give n - n - butyl - n-(12-(tert - butyldiphenylsilyloxy)dodec-5(z)-enyl)oxalamide ( 42 ) ( 1.10 g , 73% ) . 
   h nmr ( 400 mhz )  8.05 ( br s , nh , 2h ) , 7.667.74 
   ( m , 4h ) , 7.327.42 ( m , 6h ) , 5.305.42 ( m , 2h ) , 3.67 
   ( t , j = 3.9 hz , 2h ) , 3.31 ( q , j = 
   5.2 hz , 4h ) , 1.962.10 ( m , 4h ) , 1.501.64 ( m , 6h ) , 1.221.44 
   ( m , 10h ) , 1.06 ( s , 9h ) , 0.92 ( t , j = 7.8 hz , 3h ) . 
   c nmr ( 100 mhz )  160.33 , 135.80 , 134.35 , 130.73 , 129.74 , 
   129.20 , 127.83 , 64.17 , 39.89 , 39.69 , 32.79 , 31.48 , 29.94 , 29.29 , 29.07 , 
   27.46 , 27.23 , 27.14 , 27.0 , 25.96 , 20.29 , 19.46 , 13.96 . 
   hrms calcd 
   for c34h53n2o3si [ m + 
   1 ] 565.3826 , found 565.3824 . 
   n - n - butyl - n-(12-(tert - butyldiphenylsilyloxy)dodec-5(z)-enyl)oxalamide ( 42 ) ( 1.20 g , 2.12 mmol ) 
   was desilylated as described above for 39 to give n - n - butyl - n-(12-hydroxydodec-5(z)-enyl)oxalamide 
   ( 43 ) ( 0.568 g , 82% ) as a colorless solid , mp 102.8102.9 
   c . 
   h nmr ( 400 mhz )  7.69 ( br s , 2h ) , 5.205.35 
   ( m , 2h ) , 3.56 ( t , j = 4.2 hz , 2h ) , 3.26 ( q , j = 5.6 hz , 4h ) , 2.17 ( br s , 1h ) , 1.952.02 ( m , 4h ) , 
   1.441.56 ( m , 6h ) , 1.201.40 ( m , 10h ) , 0.87 ( t , j = 7.2 hz , 3h ) . 
   c nmr ( 100 mhz )  160.15 , 
   130.66 , 129.21 , 62.98 , 39.80 , 39.63 , 32.93 , 31.39 , 29.77 , 29.18 , 28.95 , 
   27.26 , 27.0 , 26.88 , 25.80 , 20.18 , 13.85 . 
   n - n - butyl - n-(12-hydroxydodec-5(z)-enyl)oxalamide ( 43 ) ( 330 mg , 1.0 mmol ) was 
   brominated as described above for 40 to give n-(12-bromododec-5(z)-enyl)-n - n - butyloxalamide ( 44 ) ( 330 mg , 84% ) as a white solid , mp 46.046.3 c . 
   h nmr ( 400 mhz )  7.79 ( br s , nh , 1h ) , 7.77 
   ( br s , nh , 1h ) , 5.205.32 ( m , 2h ) , 3.32 ( t , j = 6.4 hz , 2h ) , 3.22 ( q , j = 7.2 hz , 4h ) , 
   1.902.00 ( m , 4h ) , 1.721.82 ( m , 2h ) , 1.421.56 
   ( m , 4h ) , 1.201.40 ( m , 10h ) , 0.85 ( t , j = 
   7.3 hz , 3h ) . 
   c nmr ( 100 mhz )  160.17 , 160.15 , 
   130.40 , 129.34 , 39.77 , 39.59 , 34.12 , 32.93 , 31.40 , 29.62 , 29.0 , 28.54 , 
   27.25 , 27.24 , 27.0 , 26.91 , 20.18 , 13.85 . 
   n-(12-bromododec-5(z)-enyl)-n - n - butyloxalamide ( 44 ) ( 250 mg , 0.642 mmol ) was treated 
   with potassium cyanide as described above for 41 to give n - n - butyl - n-(12-cyanododec-5(z)-enyl)oxalamide 
   ( 45 ) ( 168 mg , 78% ) as a colorless solid , mp 83.083.3 
   c . 
   h nmr ( 400 mhz )  7.45 ( br s , nh , 
   2h ) , 5.305.40 ( m , 2h ) , 3.34 ( q , j = 8.6 hz , 
   4h ) , 2.32 ( t , j = 7.6 hz , 2h ) , 1.982.08 ( m , 
   4h ) , 1.301.68 ( m , 16h ) , 0.92 ( t , j = 7.2 
   hz , 3h ) . 
   c nmr ( 100 mhz )  160.03 ( 2c ) , 130.03 , 
   129.08 , 120.10 , 39.88 , 39.42 , 31.22 , 29.40 , 28.82 , 28.60 , 28.42 , 27.07 , 
   27.06 , 26.82 , 25.54 , 20.06 , 17.01 , 13.80 . 
   hrms calcd for c19h34n3o2 [ m + 1 ] 336.2651 , 
   found 336.2650 . to a suspension of n - n - butyl - n-(12-cyanododec-5(z)-enyl)oxalamide 
   ( 45 ) 
   ( 420 mg , 1.29 mmol ) in meoh / h2o ( 4:1 ; 
   12 ml ) was added h2nohhcl ( 228 mg , 3.28 mmol ) and 
   na2co3 ( 344 mg , 3.25 mmol ) . 
   the reaction mixture was heated at 60 c for 18 h 
   then cooled to room temperature , and all volatiles were removed in 
   vacuo . 
   the residue was diluted with water ( 30 ml ) and extracted into 
   ethyl acetate ( 2  25 ml ) . 
   the combined organic extracts were 
   washed with water ( 2  10 ml ) and brine ( 10 ml ) , dried , and purified 
   via silica gel column chromatography to give n-(13-amino-13-(hydroxyimino)tridec-5(z)-enyl)-n - n - butyloxalamide ( 46 ) ( 287 mg , 62% ) as a colorless solid , 116.3116.4 c . 
   h nmr ( cd3od , 400 mhz ) 
    5.285.40 ( m , 2h ) , 3.24 ( t , j = 6.4 
   hz , 4h ) , 1.982.00 ( m , 6h ) , 1.501.60 ( m , 6h ) , 1.261.40 
   ( m , 10h ) , 0.92 ( t , j = 7.3 hz , 3h ) . 
   c nmr ( cd3od , 100 mhz )  160.55 ( 2c ) , 156.31 , 130.05 , 
   129.18 , 39.23 , 39.09 , 31.18 , 30.63 , 29.51 , 28.83 , 28.69 , 27.10 , 26.87 , 
   26.59 , 19.88 , 12.88 . 
   the influence of eicosanoids and analogues 
   on coronary vascular tone was measured by the induced changes in isometric 
   tension of bovine coronary artery rings precontracted with the thromboxane - mimetic , 
   u46619 , as previously described . 
   all assays were conducted in triplicate 
   or greater and are means 10% sd of the reported value . 
   recombinant human seh was produced in a baculovirus expression system and was purified by affinity chromatography . 
   human seh ( 1 nm ) was incubated with inhibitors ( 0.4 < 
   [ i]final < 100000 nm ) for 5 min in 25 mm bis - tris / hcl 
   buffer ( 200 ml , ph 7.0 ) at 30 c before the substrate , cyano(2-methoxynaphthalen-6-yl)methyl trans-(3-phenyl - oxyyran-2-yl]methyl carbonate ( cmnpc ; [ s]final = 5 mm ) , was added . 
   activity was assessed by measuring 
   the appearance of the fluorescent 6-methoxynaphthaldehyde product 
   ( em = 330 nm , ex = 465 nm ) at 
   30 c during a 10 min incubation ( spectramax m2 , molecular device , 
   inc . , 
   ic50s refer to the concentrations of inhibitor that reduced activity 
   by 50% and are the averages of three replicates .
summary: the cytochrome p450 eicosanoid 14,15-epoxyeicosa-5,8,11-trienoic 
   acid ( 14,15-eet ) is a powerful endogenous autacoid that has been ascribed 
   an impressive array of physiologic functions including regulation 
   of blood pressure . because 14,15-eet is chemically and metabolically 
   labile , structurally related surrogates containing epoxide bioisosteres 
   were introduced and 
   have become useful in vitro pharmacologic tools 
   but are not suitable for in vivo applications . a new generation of 
   eet mimics incorporating modifications to the carboxylate were prepared 
   and evaluated for vasorelaxation and inhibition of soluble epoxide 
   hydrolase ( seh ) . 
   tetrazole 19 ( ed50 0.18 m ) 
   and oxadiazole-5-thione 25 ( ed50 0.36 m ) 
   were 12- and 6-fold more potent , respectively , than 14,15-eet as vasorelaxants ; 
   on the other hand , their ability to block seh differed substantially , 
   i.e. , 11 vs > 500 nm . these data will expedite the development of 
   potent 
   and specific in vivo drug candidates .