Patent Application: US-201214003909-A

Abstract:
the present invention relates to π - conjugated fluoroionophores , methods for their preparation , and their use and a method for determining an alkali ion . fluoroionophoric compounds of the general formula i are described ionophore - π - linker - fluorophore wherein the ionophore is an anilino containing crown ether or cryptand with one or more anilino donor moieties as electron donors , forming a stable complex with an alkali metal ion the π - linker is an aromatic or heteroaromatic conjugative linking moiety , and the fluorophore is an electron acceptor moiety . variation of the ionophoric unit offers a broad spectrum of detectable k + and na + - concentrations , ranging from high concentration around 800 mm down to very low concentrations around 3 mm . the fluoroionophores have great potential for application in fluorescent optode system based blood analyzing equipment for methods and kits for the determination of k + and na + concentrations in biological systems , either in vitro or in vivo , using embodiments of the disclosed fluoroionophores .

Description:
the term “ alkyl ” or “ lower alkyl ” as used herein refers to a straight or branched chain , saturated hydrocarbon having the indicated number of carbon atoms . for example , ( c1 - c6 ) alkyl is meant to include , but is not limited to methyl , ethyl , propyl , isopropyl , butyl , sec - butyl , tert - butyl , pentyl , isopentyl , neopentyl , hexyl , isohexyl , and neohexyl . an alkyl or lower alkyl group can be unsubstituted or optionally substituted with one or more substituents . the term “ alkoxy ” or “ lower alkoxy ” as used herein refers to an — o - alkyl group having the indicated number of carbon atoms . for example , a ( c1 - c6 ) alkoxy group includes — o - methyl , — o - ethyl , — o - propyl , — o - iospropyl , — o - butyl , — o - sec - butyl , — o - tert - butyl , — o - pentyl , — o - isopentyl , — o - neopentyl , — o - hexyl , — o - isohexyl , and — o - neohexyl . the term “ aromatic ” as used herein refers to an aromatic or heteroaromatic moiety . an “ aromatic ” moiety refers to a 6 - to 14 - membered monocyclic , bicyclic or tricyclic aromatic hydrocarbon ring system . examples of an aromatic group include phenyl and naphthyl . an aromatic group can be unsubstituted or optionally substituted with one or more substituents . the term “ heteroaromatic ” as used herein refers to an aromatic heterocyclic ring of 5 to 14 members and having at least one heteroatom selected from nitrogen , oxygen and sulfur , and containing at least 1 carbon atom , including monocyclic , bicyclic , and tricyclic ring systems . representative heteroaromatics are triazolyl , tetrazolyl , oxadiazolyl , pyridyl , furyl , benzofuranyl , thiophenyl , benzothiophenyl , quinolinyl , pyrrolyl , indolyl , oxazolyl , benzoxazolyl , imidazolyl , benzimidazolyl , thiazolyl , benzothiazolyl , isoxazolyl , pyrazolyl , isothiazolyl , pyridazinyl , pyrimidinyl , pyrazinyl , triazinyl , cinnolinyl , phthalazinyl , quinazolinyl , pyrimidyl , azepinyl , oxepinyl , naphthothiazolyl , quinoxalinyl . a heteroaromatic group can be unsubstituted or optionally substituted with one or more substituents . the term “ halogen ” as used herein refers to — f , — cl , — br or — i . as used herein , the term “ heteroatom ” is meant to include oxygen ( o ), nitrogen ( n ), and sulfur ( s ). as used herein , the term “ bodipy ” is an acronym for the compound class of boron - d ipyrromethanes . the new π - conjugated 1 , 2 , 3 - triazol - 1 , 4 - diylfluoroionophores according to the present invention generated via cu ( i ) catalyzed [ 3 + 2 ] cycloaddition show high fluorescence enhancement factors in the presence of na + and k + in mecn and high selectivity towards na + and k + under simulated physiological conditions . preferred embodiments of the present invention are compounds 1 and 2 as given in scheme 1 . the inventors found that in the simple cuaac - generated fluoroionophore 1 , the electronic conjugation of the n - phenylaza - 18 - crown - 6 ether and the 7 - diethylaminocoumarin fluorophore through a 1 , 2 , 3 - triazol - 1 , 4 - diyl π - linker results in a perfect signal transduction chain for the sensing of na + and k + . in mecn high cation - induced fefs were obtained for 1 ( fef na + : 58 ; fef k + : 27 ). the signal transduction in 1 also works nicely under simulated physiological conditions . in the presence of 160 mm k + a fef of 2 . 5 was observed , whereas the same concentration range of na + resulted in an almost negligible fluorescence enhancement . the inventors assume , that compound 1 is a pet - fluoroionophore with a virtual spacer between the anilinotriazole electron donor unit and the coumarin electron acceptor moiety . the cuaac of the ethinyl - functionalized n - phenylaza - 18 - crown - 6 ether 7 with 3 - azido - 7 - diethylaminocoumarin 8 afforded 1 , 2 , 3 - triazole - fluoroionophore 1 . the triazole - isomer 2 was obtained by the reaction of the azido - functionalized n - phenylaza - 18 - crown - 6 ether 9 with 3 - ethinyl - 7 - diethylamino - coumarin 10 . dye 3 , in which the crown has been replaced by a diethylamino group was synthesised in order to represent a reference compound for 1 . a further reference compound for 1 is compound 4 , in which the p - phenylen linker between aza - 18 - crown - 6 and triazole ring is replaced by a deconjugated methylene group . compound 4 was obtained by the reaction of n - propargylaza - 18 - crown - 6 ether 11 with 3 - azido - 7 - diethyl - aminocoumarin . the new 1 , 4 - disubstituted 1 , 2 , 3 - triazoles 1 - 4 are stable in solutions of mecn and dmso at room temperature for many weeks . uv - irradiation over a period of several hours did not show any decomposition of the fluoroionophores 1 and 2 . the fluorescence quantum yield of 1 in mecn is extremely low ( φ f ( 1 )= 0 . 008 , table 1 ). the fluorescence spectra of 1 in the presence of na + and k + , respectively ( fig1 ) show the impressive fluorescence enhancement upon coordination of the alkali metal ions . high fefs could be determined for na + ( fef = 58 ) and for k + ( fef = 27 ). 12 the fluorescence quantum yield of the constitutional isomer 2 in mecn is higher than that of 1 ( φ f ( 2 )= 0 . 03 , table 1 ). the fluorescence of 2 is also enhanced in the presence of increasing concentrations of na + and k + ( fig7 and 8 , respectively ). however the observed fefs for na + ( fef = 17 ) and k + ( fef = 13 ) are significantly lower than for the isomer 1 . the reference compound 4 consists of a poorer pet donor due to the aliphatic amine which is electronically separated from the triazole . hence 4 has a high quantum yield ( φ f ( 4 )= 0 . 56 , table 1 ) and is therefore less affected by the presence of na + and k + . the inventors further investigated the influence of na + and k + on the fluorescence of 1 under simulated physiological conditions . the ligand was exposed to aqueous solutions in the physiological interesting concentration range of 0 - 160 mm na + or k + , respectively . additionally , the solutions contained 2 mm ca 2 + and 2 mm mg 2 + . the ph was adjusted to 7 . 2 with 10 mm tris and a constant ionic strength of 180 mm was maintained with choline chloride . under these conditions , the fluorescence of 1 ( λex = 424 nm , λem = 500 nm , φ f = 0 . 07 ) is hardly increased in the presence of na + whereas increasing concentrations of k + resulted in a modest fluorescence enhancement ( fig2 a ). in the presence of 160 mm k + a fef of 2 . 5 is observed . the fluorescence measurements were repeated four times showing only very little variations in the fef values ( fig2 b ). it is noteworthy , that the presence of physiological important extracellular cations , such as mg 2 + and ca 2 + does not affect the signal response . in comparison , the fef of 1 for k + in mecn is clearly smaller than the fef in water under simulated physiological conditions . this can be explained by the significantly smaller stability constants of the k + complex with the n - phenylaza - 18 - crown - 6 in water [ igk ( h 2 o )& gt ; 0 . 5 ; igk ( mecn )= 3 . 95 ± 0 . 08 ]. the fact that the fluorescence of 1 in water is exclusively enhanced in the presence of k + and not by na + can be rationalized by the stronger hydration enthalpy of na + . however , the fluorescence enhancement of 1 in the presence of k + under simulated physiological conditions shows that the signaling transduction chain in this fluoroionophore works well in water . the dissociation constant ( kd ) of 1 + k + amounts to ˜ 260 mm in solutions which approximates physiological ionic strength . to measure intracellular or extracellular concentrations of k + ( kd around 140 or 4 mm ) tuning of probe 1 towards a higher complex stability while maintaining the selectivity will be necessary . to investigate the ph - sensitivity of 1 , the fluorescence intensity was measured in water at different ph values . the resulting pka of 1 is near 4 . 5 meaning that the triazole - substituted n - phenylaza - 18 - crown - 6 is less ph - sensitive than the n ( o - methoxyphenyl ) aza - 15 - crown - 5 - naphthalimide nat fluoroionophore ( pka ˜ 5 . 5 ) of the authors he et al . 13 in summary , the inventors have shown for the first time that an electronically conjugated 1 , 2 , 3 - triazole - fluoroionophore consisting of the signaling transduction chain : anilino - ionophore - 1 , 2 , 3 - triazol - 1 , 4 - diyl - fluorophore , works as an effective sensor for na + and k + in acetonitrile with high cation - induced fefs . under simulated physiological conditions 1 selectively detects k + with the modest fef being limited by the rather simple receptor unit . the inventors found that the substitution of ionophore and fluorophore on the 1 , 2 , 3 - triazole ring has a basic influence on the quality of the fluoroionophore . recently he et al . 13a and the verkman group 13b designed fluorescence switch - on pet sensors with high k + / na + selectivity and sensitivity for physiological k + in the concentration range of 0 - 40 mm . in these k + fluoroionophores a [ 3 . 2 . 2 ]- cryptand represents the ionophore . a drawback of the [ 3 . 2 . 2 ]- cryptand fluoroionophores though , is the very expensive synthetic procedure . 13c the inventors developed simple lariat aza - 18 - crown - 6 ionophores , which show a high k + / na + selectivity and sensitivity under simulated physiological conditions . these novel lariat aza - 18 - crown - 6 ionophores are also an object of the present invention . the further preferred embodiments of the compounds of the present invention containing a 3 ( 2 - methoxyethoxy ) phenyl - aza - 18 - crown - 6 as k + / na + selective and sensitive ionophore are given in scheme 3 . compounds 6 to 9 have a 2 - methoxyethoxy - lariate chain in close proximity to the aza - 18 - crown - 6 . this lariate group provides a higher binding selectivity for k . the fluorescence spectra are shown in fig4 to 7 . according to the invention na + - fluoroionophores 14 and 15 with a 1 , 2 , 3 - triazol - 1 , 4 - diyl as the π - linker ( scheme 4 ) are provided . these fluoroionophores show high na + / k + - selectivity under simuated physiological conditions . the fluorescence spectra are shown in fig8 to 11 . the following examples explain the present invention in more detail . the embodiments described in the examples are not limiting the scope of the invention . the examples should only serve as preferred embodiments and a skilled artesian will derive other embodiments from those examples without any undue burden . fluorescence titration spectra of the compounds ( c = 5 · 10 − 6 mol · l − 1 ) in acetonitrile were recorded 5 min after the addition of 0 . 02 ml of volumetric standard solution of napf 6 or kpf 6 ( c = 5 ′ 10 − 5 - 5 ′ 10 − 2 mol · l − 1 ), respectively . titration was continued until no change in fluorescence enhancement was observed . fluorescence quantum yields were determined using a pl quantum yield measurement system c9920 - 2 of hamamatsu , japan . fluorescence spectra in aqueous solutions were carried out in buffered saline solutions ( 10 mm tris - buffer , ph = 7 . 2 ) using a 1 mm ligand solution in dmso . the water used was purified by a milli - q - deioniser from millipore ®. to obtain physiological conditions , the salt solutions contained constant concentrations of cacl 2 ( 2 mm ) and mgcl 2 ( 2 mm ). the physiological solutions were varied with regard to the concentrations of respectively kcl or nacl and constant ionic strength was adjusted to a typical ionic strength in physiological systems ( 180 mm ) by choline chloride . the dye was exposed to different saline solutions containing 0 , 5 , 10 , 20 , 40 , 80 , 160 mm kcl or nacl , respectively ( table 2 and 3 ). each salt solution was mixed with 1 mm dmso solution of the compound ( 990 / 10 v / v ) to give a final ligand concentration of 10 μm . for each measurement , a freshly prepared dye mixture was used and every experiment was repeated with n = 4 . fluorescence response of the compounds was recorded exciting at the given wavelengths . all commercially available chemicals were used without further purification . solvents were distilled prior use . 1 h and 13 c nmr spectra were recorded on 300 mhz , 500 mhz or 600 mhz instruments . data are reported as follows : chemical shifts in ppm ( δ ), multiplicity ( s = singlet , d = doublet , t = triplet , m = multiplet , dd = doublet of doublets ), integration , coupling constant ( hz ). esi spectra were recorded using a micromass q - tof micro mass spectrometer in a positive electrospray mode . ir spectra were recorded using a thermo nicolet nexus ftir instrument . air / water - sensitive reactions were performed in oven - dried glassware under an argon atmosphere . column chromatography was performed with sio 2 ( merck silica gel 60 ( 0 . 04 - 0 . 063 mesh )). the preparation of 3 - azido - 7 - diethylaminocoumarine ( 60 ), 14 3 - ethinyl - 7 - diethylaminocoumarine ( 61 ), 15 n - phenylaza - 18 - crown - 6 ether 16 and n -( 4 - formyl ) phenylaza - 18 - crown - 6 ether 17 has been described previously . the preparation of 27 followed the literature and references therein according to a modified procedure . 18 a suspension of zinc ( 1 . 69 g , 25 . 9 mmol ) and carbon tetrabromide ( 8 . 59 g , 25 . 9 mmol ) in dry ch 2 cl 2 ( 40 ml ) was cooled to − 15 ° c . with an ice - salt bath before a solution of triphenylphosphine ( 6 . 79 g , 25 . 9 mmol ) in dry ch 2 cl 2 was added dropwise . the resulting yellow green mixture was kept at − 15 ° c . for 30 min and was then stirred at rt for 3 h whereupon a solution of 25 ( 4 . 31 g , 11 . 7 mmol ) in dry ch 2 cl 2 was added dropwise . the red brown mixture was stirred at rt for 2 h and then water ( 100 ml ) was added . the organic phase was separated and washed with water ( 3 × 75 ml ). the combined organic layer were dried with mgso 4 and concentrated to give 26 as a light brown oil that was directly used for the next step . note that the product decomposes when concentrated to dryness resulting in a green solid . storage at − 20 ° c . is recommended . hrms ( + esi ): m / z calcd for ( m + h ) + , 522 . 04 , 524 . 05 , 526 . 04 . found , 522 . 11 , 524 . 11 , 526 . 11 . 26 ( 2 . 91 g , 5 . 56 mmol ) was dissolved in 25 ml dry thf and set under argon atmosphere immediately . it was cooled to − 78 ° c . before n - butyllithium ( 7 . 5 ml , 1 . 6 m , solution in hexane ) was added via a syringe . after stirring for 1 h at − 78 ° c . the solution was allowed to warm up to room temperature and stirring was continued at this temperature for 1 h before h 2 o ( 15 ml ) was added cautiously . the reaction mixture was extracted with 3 × ch 2 cl 2 and the combined organic layers were dried with mgso 4 and concentrated to give 27 as a red brown oil , which was purified via chromatography ( silica gel , chcl 3 / meoh , 95 / 5 v / v ). yield : 20 %, overall . 1 h nmr ( cdcl 3 , 300 mhz ): δ = 2 . 93 ( s , 1h ), 3 . 6 ( m , 24h ), 6 . 56 ( d , 2h , j = 9 . 23 hz ), 7 . 28 ( d , 2h , j = 8 . 85 hz ); 13 c nmr ( cdcl 3 , 75 mhz ): δ = 51 . 66 , 68 . 94 , 71 . 15 , 75 . 12 , 85 . 17 , 108 . 82 , 111 . 62 , 133 . 73 , 148 . 45 ; hrms ( + esi ): m / z calcd for ( m + h ) + , 364 . 21 . found , 364 . 26 ; ir ( atr , cm − 1 ): 718 , 692 , 1115 , 2097 , 2868 , 3054 . the preparation of the n -( 4 - nitro ) phenylaza - 18 - crown ether and subsequent hydrogenation to the corresponding amine followed the literature . 19 the synthesis of 28 followed the literature according to a modified procedure . 20 n - anilino - 4 - aza - 18 - crown - 6 ether ( 4 . 03 g , 11 . 4 mmol ) was dissolved in 90 ml hcl ( 4m ) and cooled to 0 ° c . a solution of nano 2 ( 0 . 784 g , 11 . 4 mmol ) in 45 ml h 2 0 was added dropwise . the mixture was stirred for 10 min before a solution of nan 3 ( 1 . 1 g , 17 mmol ) in 45 ml h 2 o was added drop wise . stirring was continued for another 10 min at 0 ° c . before the orange solution was allowed to warm up to room temperature and it was stirred for 14 h at ambient temperature . the reaction mixture was brought to ph = 7 with na 2 co 3 , was extracted with 3 × 180 ml chcl 3 and the combined organic layers were dried with mgso 4 . the solvent was removed under reduced pressure to yield 28 as brown oil ( 1 . 31 g , 30 . 3 %). the product was used for the next reaction step without further purification . 1 h nmr ( cdcl 3 , 300 mhz ): δ = 3 . 57 - 3 . 69 ( m , 24h ), 6 . 68 ( d , 2h , j = 8 . 854 hz ), 6 . 88 ( d , 2h , j = 9 . 042 hz ); 13 c nmr ( cdcl 3 , 75 mhz ): δ = 51 . 48 , 68 . 74 , 70 . 70 , 112 . 97 , 119 . 94 , 127 . 55 , 145 . 98 ; hrms ( + esi ): m / z calcd for ( m + h ) + , 381 . 21 . found , 381 . 34 , calcd for ( m - n 2 ) + 353 . 21 . found , 353 . 30 ; ir ( atr , cm − 1 ): 1100 , 1508 , 2120 , 2097 , 2867 . the synthesis followed the literature according to a modified procedure . 21 monoaza - 18 - crown - 6 ether 22 ( 0 . 6 g , 2 . 28 mmol ) and propargylbromide ( 0 . 206 ml , 2 . 74 mmol ) were dissolved in dry acetonitrile ( 70 ml ) before cs 2 co 3 ( 1 . 48 g , 4 . 56 mmol ) was added . the suspension was stirred overnight at 85 ° c . after cooling to room temperature , the suspension was filtered and the filtrate was concentrated . the residue was purified by column chromatography on silica gel eluting with chcl 3 / meoh ( 9 / 1 v / v ) yielding 29 as a red oil ( 64 %) 1 h nmr ( cdcl 3 , 300 mhz ): δ = 2 . 18 ( t , 1h , j = 2 . 45 hz ), 2 . 82 ( t , 4h , j = 5 . 28 hz ), 3 . 61 - 3 . 74 ( m , 22h ); 13 c ( cdcl 3 , 75 mhz ): δ = 43 . 42 , 53 . 29 , 68 . 94 , 70 . 52 , 73 . 61 , 78 . 85 ; ir ( atr , cm − 1 ): 1115 , 2867 , 3189 ; hrms ( + esi ): m / z calcd . for ( m + h ) + , 302 . 20 . found , 302 . 21 . the synthesis followed the literature according to a modified procedure . 23 a mixture of zinc ( 4 . 9 g , 75 mmol ), triphenylphosphine ( 19 . 67 g , 75 mmol ), and carbon tetrabromide ( 24 . 87 g , 75 mmol ) in dry dichloromethane ( 250 ml ) was sonicated for 2 h while cooling with an ice bath . n , n - diethylaminobenzaldehyde ( 30 ) ( 5 . 30 g , 29 . 9 mmol ) was added to the grayish suspension and it was stirred overnight resulting in a brownish suspension . the mixture was concentrated and petroleum ether ( 500 ml ) was added , whereupon a tarry precipitate formed . it was washed with 1 / 1 ch 2 cl 2 / light petroleum ether ( 2 × 100 ml ). the combined organic phases were concentrated , and the residue was chromatographed on silica gel ( 150 g ), eluting with 1 / 1 ch 2 cl 2 / hexane to give 31 as a yellow oil ( 7 . 3 g , 21 . 0 mmol , 70 %). the intermediate product was kept at − 20 ° c . until the next step . 1 h nmr ( cdcl 3 , 300 mhz ): δ = 1 . 17 ( t , 6h , j = 6 . 94 hz , ch 3 ), 3 . 37 ( q , 4h , j = 6 . 94 hz , ch 2 ), 6 . 62 ( d , 2h , j = 7 . 88 hz , ar — h ), 7 . 32 ( s , 1h , ch ), 7 . 49 ( d , 2h , j = 8 . 21 hz ); hrms ( + esi ): m / z calcd . for ( m + h ) + , 331 . 96 , 333 . 96 , 335 . 96 . found , 331 . 93 , 333 . 93 , 335 . 94 . a solution of 31 ( 4 . 3 g , 12 . 4 mmol ) in dry thf ( 93 ml ) was cooled to − 78 ° c . before n - butyllithium ( 19 ml , 15 % in hexane ) was added dropwise . it was stirred for 45 min at this temperature before the reaction mixture was allowed to warm up to rt . after stirring for 1 h at rt , water ( 7 ml ) was added slowly . the solvent was removed and the residue was taken up in diethyl ether . the organic layer was washed with water ( 30 ml ) and brine ( 30 ml ) and dried over mgso 4 . concentration of the ether phase gave 32 as orange oil that was used in the next step without further purification ( 1 . 29 g , 6 . 96 mmol , 56 . 2 %). 1 h nmr ( cdcl 3 , 300 mhz ): δ = 1 . 16 ( t , 6h , j = 7 . 25 hz ), 2 . 97 ( s , 1h ), 3 . 37 ( q , 4h , j = 7 . 25 hz ,), 6 . 57 ( d , 2h , j = 9 . 14 hz ), 7 . 34 ( d , 2h , j = 9 . 14 hz ), 13 c ( cdcl 3 , 75 mhz ): δ = 12 . 46 , 44 . 26 , 74 . 41 , 85 . 01 , 107 . 47 , 110 . 94 , 133 . 39 , 147 . 73 , hrms ( + esi ): m / z calcd . for ( m + h ) + , 174 . 13 . found , 174 . 16 . into a stirred mixture of 860 mg ( 2 . 43 mmol ) aldehyd 37 and 673 mg ( 4 . 86 mmol ) k 2 co 3 in 35 ml dry methanol was added 564 mg ( 2 . 93 mmol ) dimethyl - 1 - diazo - 2 - oxopropylphosphonate ( bestmann - ohira reagent ). this mixture was stirred for 20 hours at ambient temperature . after addition of 60 ml chcl 3 and extraction 3 times with water , the organic layer was separated , dried with mgso 4 and concentrated in vacuum . the residue was purified by column chromatography on silica with chcl 3 / ch 3 oh ( 95 / 5 ) as eluent . the product was obtained as a yellow oil ( 435 mg , 51 %). 1 h nmr ( 300 mhz , cdcl 3 ): δ = 2 . 99 ( s , 1h ), 3 . 44 - 3 . 48 ( m , 4h ), 3 . 58 - 3 . 66 ( m , 16h ), 3 . 77 ( s , 3h ), 13 c nmr ( 75 mhz , cdcl 3 ): δ = 52 . 85 , 55 . 37 , 69 . 94 , 70 . 25 , 70 . 34 , 70 . 85 , 75 . 57 , 84 . 15 , 113 . 94 , 115 . 13 , 119 . 06 , 125 . 21 , 140 . 93 , 151 . 32 ir ( atr , cm − 1 ): 2923 ( s ), 2855 ( s ), 2102 ( m ), 1507 ( s ), 1253 ( s ), 1110 ( s ). elemental analysis (%) calcd . for c 19 h 27 no 5 ( 349 . 42 ): c , 65 . 31 , h , 7 . 79 , n , 4 . 01 . found c , 65 . 12 ; h , 7 . 38 ; n , 3 . 98 the synthesis of 43 followed the literature according to a modified procedure . 25 the appropriate aminocrownether ( 42 ) ( 807 mg , 2 . 37 mmol ) was dissolved in 18 ml 4 m hcl and cooled . at 0 ° c . was slowly added a solution of 163 . 6 mg ( 2 . 37 mmol ) sodium nitrite in 10 ml water . the solution was stirred for 10 minutes at 0 ° c . then was added a solution of 230 mg ( 3 . 53 mmol ) sodium azide in 10 ml water . the resulting solution was stirred for further 10 min at 0 ° c . and led warm up to room temperature . it was stirred overnight . the mixture was brought to ph 7 with potassium carbonate and extracted 3 times with chcl 3 . the combined organic layers were dried with mgso 4 and concentrated in vacuum . the residue was purified by column chromatography on silica with chcl 3 / ch 3 oh ( 95 / 5 ) as eluent . the product was obtained as a darkbrown oil ( 265 mg , 31 %). 1 h nmr ( 300 mhz , cdcl 3 ): δ = 3 . 39 - 3 . 4 ( m , 4h ), 3 . 60 - 3 . 68 ( m , 16h ), 3 . 79 ( s , 3h ) 13 c nmr ( 75 mhz , cdcl 3 ): δ = 53 . 13 , 55 . 47 , 70 . 08 , 70 . 37 , 70 . 48 , 70 . 93 , 103 . 25 , 110 . 61 , 122 . 16 , 133 . 87 , 137 . 25 , 153 . 89 ms ( ei ) m / z (%): 366 ( 12 ) [ m ] + , 338 ( 100 )[ m - n 2 ] + . ir ( atr , cm − 1 ): 2858 ( s ), 2102 ( s ), 1504 ( s ), 1234 ( s ), 1105 ( s ). the reaction followed the literature according to a modified procedure . 26 in a dry 250 - ml round bottom flask , under a stream of argon gas , were placed the 4 - bromo - 1 , 8 - naphthalic anhydride ( 95 w %, 1 . 35 g , 4 . 63 mmol , 1 equiv .) and 1 - butylamine ( 99 w %, 410 . 35 mg , 5 . 5 mmol , 1 . 2 equiv .) in dry ethanol ( 96 ml ) and the reaction mixture was heated at 60 ° c . for 16 h . after cooling to room temperature , the solid was filtered and washed with 200 ml h 2 o to give the product as a light yellow solid . yield : 0 . 800 mg , 52 %, mp : 100 - 102 ° c . ( lit . : 103 - 105 ° c ., r f = 0 . 8 ( n - hexane / ethyl acetate = 4 : 1 v / v ); 1 h nmr , ( cdcl 3 , 300 mhz ) 6 ( ppm ), j ( hz ): 8 . 63 ( d , 1h , j = 7 . 29 , h 1 ), 8 . 53 ( d , 1h , j = 8 . 52 , h 3 ), 8 . 39 ( d , 1h , j = 7 . 86 , h 4 ), 8 . 01 ( d , 1h , j = 7 . 9 , h 5 ), 7 . 82 ( tr , 1h , j = 8 . 46 , h 2 ), 4 . 16 ( t , 2h , j = 7 . 55 , h 6 ), 1 . 71 ( qu , 2h , j = 7 . 5 , h 7 ), 1 . 45 ( sx , 2h , j = 7 . 5 , h 8 ), 0 . 97 ( t , 3h , j = 7 . 32 , h 9 ); 13 c nmr , ( cdcl 3 , 75 mhz ) δ ( ppm ): 163 . 74 , 163 . 71 , 133 . 30 , 132 . 11 , 131 . 30 , 131 . 21 , 130 . 74 , 130 . 28 , 129 . 12 , 128 . 19 , 123 . 30 , 122 . 45 , 40 . 53 , 30 . 31 , 20 . 52 , 13 . 98 ; a dry 100 - ml two - necked round - bottom flask was set under argon atmosphere before n - n - butyl - 4 - bromo - 1 , 8 - naphthalimide ( 0 . 8 g , 2 . 4 mmol , 1 equiv . ), thf ( 8 ml ), pd ( pph 3 ) 2 cl 2 ( 33 . 8 mg , 0 . 048 mmol , 0 . 02 equiv . ), pph 3 ( 25 . 3 mg , 0 . 096 mmol , 0 . 04 equiv . ), cui ( 18 . 3 mg , 0 . 096 mmol , 0 . 04 equiv .) and dry tea ( 8 ml ) were added followed by the trimethylsilylacetylene ( 331 . 1 mg , 3 . 37 mmol , 1 . 4 equiv .). it was refluxed for 6 h . then the reaction mixture was allowed to cool down to room temperature and the solvent was evaporated under reduced pressure . the crude product was purified by column chromatorgraphy ( 50 × 3 cm , silica gel , ch 2 cl 2 = 10 : 1 v / v ) to give the productas a yellow solid . yield : 0 . 780 g , 92 %, mp : 136 . 2 - 138 . 8 ° c ., r f = 0 . 87 ; 1 h nmr , ( cdcl 3 , 300 mhz ) 6 ( ppm ), j ( hz ): 8 . 61 ( dd , 2h , h 4 , h 3 ), 8 . 50 ( d , 1h , j = 7 . 62 , h 1 ), 7 . 88 ( d , 1h , j = 7 . 62 , h 5 ), 7 . 8 ( t , 1h , j = 7 . 62 , h 2 ), 4 . 17 ( t , 2h , j = 7 . 5 , h 6 ), 1 . 71 ( qu , j = 7 . 5 , 2h , h 7 ), 1 . 37 - 1 . 51 ( m , 2h , j = 7 . 53h 8 ), 0 . 97 ( t , 3h , j = 7 . 3 , h 9 ), 0 . 36 ( s , 9h , h 10 ); 13 c nmr , ( cdcl 3 , 75 mhz ) δ ( ppm ): 164 . 14 , 163 . 86 , 132 . 49 , 131 . 93 , 131 . 68 , 131 . 32 , 130 . 35 , 128 . 07 , 127 . 63 , 127 . 37 , 123 . 11 , 122 . 53 , 105 . 37 , 101 . 44 , 40 . 48 , 30 . 36 , 20 . 54 , 13 . 99 , 0 . 002 ; n - n - butyl - 4 -( trimethylsilylethinyl )- 1 , 8 - naphthalimide ( 710 mg , 2 . 03 mmol ) was placed into a 100 - ml round - bottom flask and in dry methanol ( 50 ml ) and dry k 2 co 3 ( 1 . 09 g , 7 . 92 mmol , 3 . 9 equiv .) were added . it was heated for 16 h at room temperature . ch 2 cl 2 ( 40 ml ) was added to the reaction mixture and the organic phase was washed with water ( 2 × 20 ml ). the combined organic phases were dried over na 2 so 4 and concentrated in vacuo . the crude product was purified by column chromatography ( 50 × 3 cm , silica gel , ch 2 cl 2 / ethyl acetate = 10 : 1 v / v ) to give the product as a yellow solid . yield : 0 . 200 g , 35 %, mp . : 127 . 5 - 130 . 8 ° c ., r f = 0 . 81 ; 1 h nmr , ( cdcl 3 , 600 mhz ) 6 ( ppm ), j ( hz ): 8 . 65 ( d , 1h , j = 8 . 43 , h 3 ), 8 . 63 ( d , 1h , j = 7 . 32 , h 4 ,), 8 . 53 ( d , 1h , j = 7 . 56 , h 1 ), 7 . 93 ( d , 1h , j = 7 . 56 , h 5 ), 7 . 82 ( t , 1h , j = 8 . 25 , h 2 ), 4 . 17 ( t , 2h , j = 7 . 62 , h 6 ), 3 . 73 ( s , 1h , h 10 ), 1 . 71 ( qu , 2h , j = 7 . 56 , h 7 ), 1 . 44 ( sx , 2h , j = 7 . 56 , h 8 ), 0 . 97 ( t , 3h , j = 7 . 38 , h 9 ); 13 c nmr , ( cdcl 3 , 75 mhz ) δ ( ppm ): 164 . 05 , 163 . 78 , 132 . 24 , 132 . 12 , 131 . 77 , 130 . 245 , 128 . 09 , 127 . 81 , 126 . 30 , 123 . 24 , 123 . 07 , 122 . 01 , 86 . 52 , 80 . 51 , 40 . 50 , 30 . 36 , 20 . 52 , 13 . 95 ; the synthesis followed the literature according to a modified procedure .′ 1b a suspension of 4 - bromo - 1 , 8 - naphthalic anhydride ( 2 g , 7 . 2 mmol , 1 equiv .) and ethylamine ( 70 w % in water , 0 . 69 ml , 8 . 66 mmol , 1 . 2 equiv .) in 100 ml 1 . 4 - dioxane was refluxed for 6 h giving a brownish solution . the solution was cooled to room temperature before it was poured onto 300 ml ice water resulting in an off white precipitate . after complete melting of the ice , the precipitate was collected by filtration and was washed with water . the resulting solid was dried in vacuo in a desiccator equipped with cacl 2 to give the product as an off white solid which was directly used in the next step . yield : 1 . 72 g , 78 %, mp : 145 - 146 ° c ., 1 h nmr , ( cdcl 3 , 500 mhz ) 6 ( ppm ), j ( hz ): 8 . 62 ( dd , 1h , h 1 ), 8 . 34 ( dd , 1h , h 3 ), 8 . 37 ( d , 1h , j = 7 . 85 , h 4 ), 8 . 0 ( d , 1h , j = 7 . 80 , h 5 ), 7 . 81 ( tr , 1h , j = 7 . 35 , h 2 ), 4 . 22 ( q , 2h , j = 7 . 15 , h 6 ), 1 . 32 ( tr , 3h , j = 7 . 10 , h 7 ); ms ( ei ): m / z calcd . : 303 . found , 303 + 305 : [ m / z - ch 2 ch 3 ] + calcd . : 275 . found 275 + 277 ; the synthesis followed the literature and references therein . in a 25 - ml round - bottom flask was placed the n - n - ethyl - 4 - bromo - 1 , 8 - naphthalimide ( 0 . 5 g , 1 . 64 mmol , 1 equiv .) and sodium azide ( 0 . 53 g , 8 . 2 mmol , 5 equiv .). then n - methylpyrrolidinone ( 7 ml ) was added and the mixture was heated at 110 ° c . for 1 . 5 h . the solution was allowed to cool to room temperature and diluted with 25 ml water . it was extracted with ethyl acetate ( 3 × 20 ml ) and the combined organic phases were washed with brine . the organic phase was dried over mgso 4 and concentrated under reduced pressure . the crude product was purified by column chromatography ( silica gel , hexane / ethyl acetate = 4 : 1 v / v ) to give the product as a yellow solid . yield : 0 . 254 g , 58 %, 1 h nmr , ( cdcl 3 , 300 mhz ) 6 ( ppm ), j ( hz ): 8 . 62 ( dd , 1h , h 1 ), 8 . 56 ( dd , 1h , h 4 ), 8 . 41 ( dd , 1h , h 3 ), 7 . 72 ( m , 1h , h 2 ), 7 . 45 ( dd , 1h , h 5 ), 4 . 23 ( q , 2h , j = 7 . 14 , h 6 ), 1 . 32 ( tr , 3h , j = 7 . 08 , h 7 ); 13 c nmr , ( cdcl 3 , 75 mhz ) δ ( ppm ): 163 . 92 , 163 . 51 , 143 . 50 , 132 . 26 , 131 , 74 , 129 . 27 , 128 . 82 , 126 . 96 , 124 . 49 , 122 . 85 , 119 . 15 , 114 . 78 , 35 . 65 , 13 . 48 ; ms ( ei ): m / z calcd . : 266 . found , 266 ; [ m / z - n 2 ] + calcd . : 238 . found 238 ; ir ( kbr , cm − 1 ): 2131 ( s , n 3 ). yield : 78 %, 1 h nmr , ( cdcl 3 , 300 mhz ) 6 ( ppm )): 8 . 58 ( dd , 2h , h 1 ), 8 . 46 ( s , 1h , h 5 ), 8 . 02 ( dd , 2h , h 4 ), 8 . 62 - 8 . 45 ( m , 4h , h ″), 3 . 99 ( s , 1h , h 6 ) the synthesis followed the literature according to a modified procedure . 29 a suspension of chloromethylanthracene ( 1 g , 4 . 41 mmol , 1 equiv .) and sodium azide ( 0 . 43 g , 6 . 62 mmol , 1 . 5 equiv .) in 30 ml acetonitrile was refluxed for 5 h . after the reaction was complete ( monitoring via dc ) the reaction mixture was cooled to room temperature and the resulting solid was filtered off . the solution was concentrated in vacuo and the resulting yellow solid was purified by column chromatography on silica gel ( n - hexane / ethyl acetate = 3 : 1 v / v ). yield : 0 . 92 g ( 90 %), mp . : 78 - 80 ° c ., 1 h - nmr , ( cdcl 3 , 500 mhz ) 6 ( ppm ), j ( hz ): 8 . 5 ( s , 1h , h 5 ), 8 . 29 ( d , 2h , j = 8 . 85 , h 1 ), 8 . 05 ( d , 2h , j = 8 . 40 , h 4 ), 7 . 60 ( m , 2h , h 2 ), 7 . 52 ( m , 2h , h 3 ), 5 . 31 ( s , 2h , h 6 ); 13 c - nmr , ( cdcl 3 , 125 mhz ) δ ( ppm ): 131 . 45 , 130 . 79 , 129 . 39 , 129 . 09 , 126 . 94 , 125 . 87 , 125 . 30 , 123 . 31 , 46 . 42 ; ms ( ei ): m / z calcd . : 233 . found , 233 ; ft - ir ( kbr ), cm − 1 : 2095 ( s , — n 3 ), 1444 ( m , — ch 2 —). the product was collected after crystallization from acetic anhydride in form of yellow needles . yield : 81 %, mp : 207 - 208 ° c . ; 1 h - nmr , ( cdcl 3 , 300 mhz ) 6 ( ppm ), j ( hz ): 8 . 38 ( d , 2h , j = 8 . 61 , h 1 ), 8 . 29 ( d , 2h , j = 8 . 79 , h 4 ), 7 . 69 - 7 . 5 ( m , 4h , h 2 , 3 ), 3 . 09 ( s , 3h , h 5 ); 13 c - nmr , ( cdcl 3 , 75 mhz ) δ ( ppm ): 138 . 24 , 132 . 98 , 129 . 51 , 128 . 45 , 126 . 29 , 126 . 17 , 125 . 48 , 117 . 83 , 104 . 41 , 14 . 91 ; hrms ( + esi ): m / z calcd for ( m + h ) + , 218 . 10 . found , 218 . 15 . recrystallization from n - hexanes / chcl 3 at − 20 ° c . gave the product as a yellow solid ( needles ). 1 h - nmr , ( cdcl 3 , 300 mhz ) 6 ( ppm ), j ( hz ): 8 . 49 - 8 . 43 ( m , 2h , h 1 ), 8 . 39 - 8 . 31 ( m , 2h , h 4 ), 7 . 77 - 7 . 69 ( m , 4h 2 , 3 ), 5 . 46 ( s , 2h , h 5 ); 13 c - nmr , ( cdcl 3 , 75 mhz ) δ ( ppm ): 135 . 07 , 133 . 18 , 128 . 95 , 128 . 85 , 127 . 68 , 126 . 45 , 124 . 42 , 117 . 20 , 107 . 84 , 24 . 97 ; ft - ir ( kbr , cm − 1 ): 3051 ( w , br — ch 2 —), 2212 ( s , — c ≡ n ), 1444 ( m , — ch 2 —). the synthesis followed the procedure of the preparation of 9 -( azidomethyl )- anthracene as described above . the crude product was recrystallized from ch 2 cl 2 / meoh to give the product as yellow fluffy needles . yield : 39 %, r f = 0 . 41 ; 1 h - nmr , ( cdcl 3 , 300 mhz ) 6 ( ppm ), j ( hz ): 8 . 48 ( d , 2h , j = 8 . 84 , h 1 ), 8 . 35 ( d , 2h , j = 8 . 13 , h 4 ), 7 . 79 - 7 . 65 ( m , 4h , h 2 , 3 ), 5 . 33 ( s , 2h , h 5 ); 13 c - nmr , ( cdcl 3 , 75 mhz ) δ ( ppm ): 133 . 04 , 132 . 90 , 129 . 94 , 128 . 85 , 127 . 84 , 126 . 52 , 124 . 48 , 117 . 07 , 108 . 08 , 46 . 29 ; hrms ( + esi ): m / z calcd for ( m + h ) + , 259 . 10 . found , 259 . 15 ; ft - ir ( kbr ) u ( cm − 1 ): 2213 ( s , — c ≡ n ), 2100 ( s , — n 3 ), 1444 ( m , — ch 2 —). a 500 - ml round - bottom flask , equipped with condenser , thermometer and stirbar was set under argon atmosphere and the anthraquinone ( 10 . 5 g , 0 . 050 mol ) and dry thf ( 375 ml ) were added . the mixture was cooled to − 78 ° c . before a solution of methyl lithium ( 110 ml , 1 . 4 m in diethylether , 0 . 151 mol , 3 . 05 equiv .) was added cautiously via a syringe . the reaction mixture was slowly allowed to warm up to room temperature and was stirred at this temperature for 2 h before water ( 100 ml ) was added . the mixture was extracted with ch 2 cl 2 ( 2 × 100 ml ) and the combined organic phases were concentrated to give the 9 , 10 - dihydroxy - 9 , 10 - dimethylanthracene as a white solid which was used in the next step without further purification . it was dissolved in thf ( 250 ml ) and a solution of thf ( 130 ml ) and hbr ( 150 ml , 48 w %) was added drop wise . the reaction mixture was stirred at ambient temperature for 30 min where upon yellow crystals formed . the crystals were collected by filtration , washed with water and dried under vacuum to give the final product . yield : 10 . 16 g , 72 %, mp : 177 . 5 - 179 . 0 ° c ., r f = 0 . 25 ( chcl 3 / meoh = 97 : 3 v / v ); 1 h - nmr , ( cdcl 3 , 300 mhz ) 6 ( ppm ), j ( hz ): 8 . 35 ( t , 4h , j = 7 . 56 , h 1 , 5 ), 7 . 65 ( tr , 2h , j = 6 . 9 , j = 8 . 2 , h 3 ), 7 . 54 ( tr , 2h , j = 6 . 9 , j = 8 . 2 , h 4 ), 5 . 55 ( s , 2h , h 1 ), 3 . 09 ( s , 3h , h 6 ); 13 c - nmr , ( cdcl 3 , 75 mhz ) δ ( ppm ): 133 . 30 , 130 . 28 , 129 . 58 , 126 . 39 , 126 . 32 , 125 . 67 , 125 . 39 , 124 . 26 , 27 . 97 , 14 . 74 ; hrms ( + esi ): m / z calcd for ( m + h ) + , 286 . 19 . found , 286 . 32 ; the synthesis followed the procedure of the preparation of 9 -( azidomethyl )- anthracene as described above . yield : 90 %, r f = 0 . 32 ; 1 h - nmr , ( cdcl 3 , 300 mhz ) 6 ( ppm ), j ( hz ): 8 . 39 - 8 . 31 ( m , 4h , h 2 , 5 ), 7 . 63 - 7 . 53 ( m , 4h , h 3 , 4 ), 5 . 34 ( 5 , 2h , h 1 ), 3 . 14 ( 5 , 3h , h 6 ); 13 c - nmr , ( cdcl 3 , 75 mhz ) δ ( ppm ): 132 . 91 , 130 . 62 , 130 . 03 , 126 . 44 , 125 . 74 , 125 . 22 , 124 . 29 , 46 . 69 , 14 . 64 ; ms ( ei ): m / z : 247 ; [ m / z -. n 2 ]= 218 ; [ m / z -. ch 2 n 3 ]= 205 ; ft - ir ( kbr ), cm − 1 : 2214 ( s , c ≡ n ), 2114 ( 5 , n 3 ), 1444 ( m , ch 2 ). the synthesis followed the literature according to a modified procedure . 32 a mixture of n -( 2 - methoxyethoxy )- 2 - nitrobenzole 33 ( 21 . 9 g ; 0 . 131 mol ; 1 equiv .). 2 - chlorethanol ( 52 . 77 g ; 0 . 656 mol ; 5 equiv .) and caco 3 ( 18 . 37 ; 0 . 184 mol ; 1 . 4 equiv .) in dist . water ( 300 ml ) was stirred for 6 days at 60 ° c . after cooling to room temperature , na 2 co 3 ( 75 . 0 g ; 0 . 7 mol ; 5 . 34 equiv .) was added and it was stirred for 40 min at 60 ° c . after which the solid is filtered off . the aqueous phase is extracted tertbutyl - methyl - ether ( 3 × 500 ml ). the combined organic phases are dried over mgso 4 and concentration in vacuo gave the crude product as a brownish oil , which was purified by column chromatography ( silica gel . ethyl acetate ). yield : ( 30 . 1 g . 90 %). 1 h nmr ( cdcl 3 . 300 mhz ): δ = 3 . 15 ( tr . 4h . h 6 ). 3 . 43 ( s . 3h . h 9 ). 3 . 47 ( tr . 4h . h 5 ). 3 . 74 ( tr . 2h . h 8 ). 4 . 11 ( tr . 2h . h 7 ). 6 . 91 ( dd . 1h . h 4 ). 6 . 98 ( tr . 1h . h 2 ). 7 . 11 ( tr . 1h . h 3 ). 7 . 22 ( d . 1h . h 1 ); 13 c nmr ( cdcl 3 . 75 mhz ): δ = 57 . 93 ; 58 . 93 ; 59 . 66 ; 67 . 80 ; 70 . 70 ; 113 . 33 ; 122 . 23 ; 125 . 51 ; 126 . 01 ; 139 . 28 ; 155 . 25 ; hrms ( + esi ): m / z calc . for ( m + h ) + . 256 . 15 . found . 256 . 13 . the synthesis followed the literature according to a modified procedure . 34 the n , n - bis ( 2 - hydroxyethyl )- 2 - methoxyethyloxyaniline ( 15 . 46 g ; 60 . 55 mmol ; 1 equiv .) was dissolved in 440 ml dry acetonitrile and set under argon atmosphere . under a stream of argon sodiumhydride ( 80 % ig ; 4 . 5 g ; 2 . 86 equiv .) was added to the reaction mixture over a period of 1 h . the 1 , 17 - ditosyl - 3 , 6 , 9 , 12 , 15 - pentaoxaheptadecane ( 30 . 4 g ; 60 . 55 mmol ; 1 equiv .) was dissolved in 216 ml dry acetonitrile and added dropwise to the refluxing reaction mixture over 4 h . it was heated to reflux for 11 h before the yellow suspension was allowed to cool to room temperature . it was filtered and the solvent was removed to give a brown oil . which was purified by column chromatography ( silica gel . chcl 3 . meoh , 95 / 5 , v / v ) to give the product as a light brown oil . 1 h nmr ( cdcl 3 . 300 mhz ): δ = 3 . 41 ( s . 3h . h 13 ). 3 . 45 - 3 . 66 ( m . 24h . h 5 - 10 ). 3 . 73 ( tr . 2h . j = 4 . 9 hz . h 12 ). 4 . 10 ( tr . 2h . j = 4 . 9 hz . h 13 ). 6 . 82 - 7 . 12 ( m . 4h . h 1 - 4 ); 13 c nmr ( cdcl 3 . 75 mhz ): δ = 56 . 70 ; 59 . 91 ; 59 . 83 ; 67 . 83 ; 69 . 57 - 71 . 18 ; 114 . 39 ; 121 . 24 ; 122 . 21 ; 124 . 21 ; 151 . 71 ; 153 . 9 ; hrms ( + esi ): m / z ber . für ( m + h ) + . 255 . 16 ; gefunden . 255 . 13 . the synthesis followed the literature according to a modified procedure . 35 the 2 - methoxyethoxyphenylaza - 18 - crown - 6 - ether ( 4 . 4 g ; 10 . 65 mmol ; 1 equiv .) was dissolved in a mixture of dist . water ( 340 ml ) and glacial acetic acid ( 34 ml ) before a solution of nano 2 ( 0 . 81 ; 11 . 7 mmol ; 1 . 1 equiv .) in deionized water was added drop wise over a period of 10 min . it was stirred for 16 h at room temperature . the deep orange reaction mixture was neutralized with lioh followed by extraction with ch 2 cl 2 ( 3 × 100 ml ) the combined organic phases were dried over mgso 4 and the solvent was removed in vacuo to give the crude product as an orange oil . purification by column chromatography yielded the product as an orange oil ( silica gel . ch 3 cl / meoh , 95 / 5 , v / v ) ( 1 . 1 g . 22 . 4 %). 1 h nmr ( cdcl 3 . 300 mhz ): δ = 3 . 40 ( s . 3h . h 12 ). 3 . 60 - 3 . 70 ( m . 24h . h 4 - 9 ). 3 . 73 ( m , 2h , h 11 ). 4 . 14 ( m , 2h , h 10 ). 6 . 88 ( d . 1h . j = 9 . 04 hz . h 3 ). 7 . 65 ( d , 1h . j = 2 . 5 hz , h 1 ). 7 . 80 ( dd , 1h , h 2 ); hrms ( + esi ): m / z calc . ( m + h ) + . 459 . 23 . found , 459 . 26 . 13 c nmr ( cdcl 3 . 75 mhz ): δ = 52 . 83 , 58 . 82 , 67 . 91 , 69 . 93 , 70 . 52 , 70 . 55 , 72 . 57 , 70 . 64 , 70 . 69 , 70 . 77 , 108 . 20 . 115 . 65 , 118 . 57 , 139 . 07 , 146 . 32 , 148 . 38 ; hrms ( + esi ): m / z calc . for ( m + h ) + . 459 . 23 . found . 459 . 25 . the n - 2 - methoxyethoxy - 4 - nitrophenylaza - 18 - crown - 6 - ether ( 0 . 22 g . 0 . 48 mmol ) was dissolved in dry methanol ( 30 ml ) after addition of pd / c ( 30 mg ) it was hydrated in an autoclave for 16 h at 75 bar . the catalyst was filtered of through a bed of celite ® and the solvent was removed in vacuo to yield a colorless oil ( 0 . 2 g ; 97 %). note that the compound decomposes quickly when exposed to air , resulting in a purple colour . that &# 39 ; s why it was used in the next step without further purification . hrms ( + esi ): m / z calc . for ( m + h ) + . 429 . 26 . found . 429 . 20 . due to the instability of the n - 3 -(- 2 - methoxyethoxyphenylaza - 18 - crown - 6 -) aniline . the reaction was carried out under argon atmosphere and in oven dried glassware . the aqueous solutions were degassed and flushed with argon prior use . the n - 3 -(- 2 - methoxyethoxyphenylaza - 18 - crown - 6 -) aniline ( 0 . 2 g ; 0 . 467 mmol ; 1 equiv .) was dissolved in hcl ( 3 . 6 ml ; 4m ) gelost and cooled to 0 ° c . before a solution of nano 2 ( 32 mg ; 0 . 467 mmol ; 1 equiv .) in 1 . 8 ml h 2 o was slowly added via a syringe . the reaction mixture was stirred at this temperature for 10 min . before a solution of nan 3 ( 0 . 45 mg . 0 . 7 mmol ; 1 . 5 equiv .) in 1 . 8 ml h 2 0 was added dropwise followed by 10 in of stirring at 0 ° c . the solution was allowed to warm up to room temperatur and stirred at ambient temperatuer for 14 h . the reaction mixture was neutralized with li 2 co 3 before it was extracted with 3 × 50 ml chcl 3 . the organic layers were combined , dried with mgso 4 , filtered and the solvent was removed under reduced pressure . the crude product was purified by column chromatography ( silica gel . ch 3 cl / meoh ( 95 / 5 ) yielding the desired product as light yellow oil ( 0 . 13 g . 61 . 3 %). 1 h nmr ( cdcl 3 . 300 mhz ): δ = 3 . 4 ( s . 3h . h 12 ). 3 . 52 - 3 . 68 ( m . 24h . h 4 - 9 ). 3 . 75 ( tr . 2h . j = 4 . 9 hz . h 11 ). 4 . 10 ( tr . 2h . j = 4 . 9 hz . h 10 ). 6 . 51 ( s . 1h . h 3 ). 6 . 60 ( d . 1h . j = 8 . 48 . h 2 ). 7 . 09 ( d . 1h . j = 8 . 48 hz . h 2 ); 13 c nmr ( cdcl 3 . 75 mhz ): δ = 53 . 18 . 59 . 06 . 67 . 82 . 69 . 76 . 70 . 31 . 70 . 55 . 70 . 72 . 70 . 61 . 71 . 00 . 105 . 02 . 111 . 48 . 123 . 52 . 134 . 51 . 136 . 95 . 153 . 87 ; hrms ( + esi ): m / z calc . for ( m + h ) + found . 455 . 17 . the synthesis of the 6 - ethynyl - 2 - hexyl - 1 . 8 - naphthalimidfollowed the literature according to a modified procedure . 36 the 4 - bromonaphthalene anhydride ( 1 . 0 g . 3 . 61 mmol . 1 eq .) and n - butylamine ( 1 . 04 g . 4 . 32 mmol . 1 . 2 äq .) were stirred in dry ethanol ( 96 ml ) at 60 c for 16 h . the reaction mixture was allowed to warm up to room temperature and before the solid was filtered off . it was washed with 200 ml h 2 o and dried in high vacuo to give the product as a light yellow solid ( 1 . 74 g ; 93 %) 1 h nmr ( cdcl 3 . 300 mhz ): δ = 0 . 89 ( tr . 3h . h 1 ). 1 . 25 - 1 . 48 ( m . 24h . h 2 - 4 ). 1 . 73 ( qu . 2h . h 5 ). 4 . 17 ( tr . 2h . h 6 ). 7 . 85 ( tr . 1h . j = 7 . 35 hz . h 8 ). 8 . 06 ( d . 1h . j = 7 . 72 hz . h 11 ). 8 . 57 ( d . 1h . j = 7 . 91 hz . h 10 ). 8 . 57 ( dd . 1h . h 7 ). 8 . 66 ( d . 1h . j = 7 . 35 hz . h 9 ); 13 c nmr ( cdcl 3 . 75 mhz ): δ = 14 . 50 . 23 . 08 . 27 . 52 . 28 . 47 . 29 . 75 . 29 . 95 . 30 . 00 . 30 . 03 . 30 . 05 . 30 . 06 . 30 . 09 . 32 . 32 . 41 . 02 . 122 . 69 . 123 . 55 . 128 . 42 . 129 . 34 . 130 . 50 . 130 . 96 . 131 . 43 . 131 . 52 . 132 . 33 . 133 . 50 . 163 . 90 . 163 . 92 ; ms ( ei ): m / z calcd . : 505 . found . 499 + 501 ; under argon atmosphere . to a dry flask provided with the 6 - bromo - 2 - hexadecyl - 1 . 8 - naphthalimid ( 0 . 916 g ; 1 . 83 mmol ; 1 equiv .). pd ( pph 3 ) 2 cl 2 ( 26 . 0 mg . 0 . 037 mmol . 0 . 02 equiv .). pph 3 ( 19 . 0 mg . 0 . 037 mmol . 0 . 04 equiv .). cui ( 14 . 0 mg . 0 . 037 mmol . 0 . 04 equiv .) and dry triethylamine were added . dry thf ( 10 ml ) was transfered into the flask via a canula followed by the dropwise addition of trimethylsilylacetylene ( 0 . 252 mg . 2 . 56 mmol . 1 . 4 equiv .). the reaction mixture was heated to reflux for 6 h and then allowed to cool down to room temperature . the solvent was removed and the residue was taken up chcl 3 . water was added and it was extracted with 3 × 30 ml chcl 3 . the organic layers were combined . dried over mgso 4 and filtered . the solvent was removed in vacuo to yield the desired product quantitatively as a grey solid ( 0 . 94 g . 100 %). 1 h nmr ( cdcl 3 . 300 mhz ): δ = 0 . 35 ( s . 9h . j = 6 . 97 hz . h 12 ). 0 . 86 ( tr . 3h . h 1 ). 1 . 23 - 1 . 34 ( m . 24h . h 2 - 4 ). 1 . 71 ( qu . 2h . j = 7 . 54 hz . h 5 ). 4 . 14 ( tr . 2h . j = 7 . 72 hz . h 6 ). 7 . 80 ( tr . 1h . j = 7 . 35 . h 8 ). 7 . 89 ( d . 1h . j = 7 . 35 hz . h 7 ). 8 . 51 ( d . 1h . j = 7 . 72 hz h 11 ). 8 . 62 ( dd . 2h . h 9 , 10 ); 13 c nmr ( cdcl 3 . 75 mhz ): δ = 9 . 08 . 14 . 03 . 22 . 64 . 27 . 13 . 28 . 12 . 29 . 31 . 29 . 34 . 29 . 51 . 29 . 56 . 29 . 60 . 29 . 62 . 29 . 63 . 29 . 65 . 31 . 89 . 40 . 57 . 46 . 15 . 101 . 32 . 105 . 20 . 122 . 45 . 123 . 36 . 127 . 46 . 130 . 16 . 131 . 15 . 131 . 48 . 132 . 29 . 163 . 163 . 94 ; ms ( ei ): m / z calcd . : 517 . found . 517 ; the 2 - hexadecyl - 6 -(( trimethylsilyl ) ethynyl )- 1 . 8 - naphthalimid ( 198 mg . 0 . 383 mmol ; 1 equiv .) was dissolved in 10 ml dry methanol . dry k 2 co 3 ( 212 mg . 1 . 53 mmol . 4 equiv .) was added and it was stirred for 50 h at roomtemperature the red brown suspension was filtered and the collected solid was purified by column chromatography ( 80 × 3 cm . kieselgel . dichlormethan / essigsaureethylester = 95 : 5 v / v ) to give the pure product as a light yellow solid ( 0 . 91 g . 53 %). 1 h nmr ( cdcl 3 . 300 mhz ): δ = 0 . 87 ( tr . 3h . j = 6 . 78 hz . h 1 ). 1 . 24 - 1 . 44 ( m . 26h . h 2 - 4 ). 1 . 72 ( qu . 2h . j = 7 . 53 hz . h 5 ). 3 . 73 ( s . 1h . h 12 ). 4 . 19 ( tr . 2h . j = 7 . 53 hz . h 6 ). 7 . 82 ( tr . 1h . j = 7 . 35 hz . h 8 ). 7 . 96 ( d . 1h . j = 7 . 72 hz . h 11 ). 8 . 26 ( d . 1h . j = 7 . 53 hz . h 10 ). 8 . 64 ( dd . 2h . h 7 , 9 ); 13 c nmr ( cdcl 3 . 75 mhz ): δ = 14 . 22 . 22 . 83 . 27 . 32 . 28 . 30 . 29 . 50 . 29 . 52 . 29 . 70 . 29 . 56 . 28 . 80 . 29 . 82 . 29 . 85 . 32 . 08 . 40 . 79 . 80 . 56 . 86 . 50 . 123 . 16 . 123 . 30 . 126 . 31 . 127 . 80 . 128 . 14 . 130 . 24 . 131 . 76 . 131 . 79 . 132 . 16 . 132 . 23 . 163 . 76 . 164 . 03 ; ms ( ei ): m / z calcd . : 445 . found . 445 ; the cu ( i ) catalyzed reaction between an azide and an alkyne ( cuaac ) has been performed as follows : all reactions were performed on a mmol - scale . to a solution of azide ( 1 eq ) and alkyne ( 1 eq ) in thf / h 2 o ( 3 / 1 ) was added cui ( 5 mol %) and sodium ascorbate ( 2 . 5 mol %). it was stirred overnight at 50 ° c . the thf was removed under reduced pressure and the residue was taken up in chcl 3 . it was washed with water and the organic phase was dried over mgso 4 . the organic phase was concentrated and the residue was chromatographed on silica gel eluting with ch 2 cl 2 / meoh ( 95 / 5 ). the synthesis followed the general procedure of the cuaac reaction . yield : 49 %, 1 h nmr ( cdcl 3 , 500 mhz ): δ = 1 . 23 ( t , 6h , j = 7 . 25 hz ), 3 . 44 ( q , 4h , j = 7 . 25 hz ,), 3 . 64 - 3 . 67 ( m , 20h ), 3 . 72 ( t , 4h , j = 5 . 68 hz ), 6 . 55 ( s , 1h ), 6 . 66 ( dd , 1h ), 6 . 75 ( d , 2h , j = 8 . 52 hz ), 7 . 41 ( d , 1h , j = 8 . 83 hz ), 7 . 74 ( d , 2h , j = 8 . 83 hz ), 8 . 15 ( s , 1h ), 8 . 38 ( s , 1h ); 13 c ( cdcl 3 , 75 mhz ): δ = 13 . 53 , 46 . 05 , 52 . 38 , 69 . 78 , 71 . 89 , 98 . 08 , 108 . 27 , 110 . 91 , 112 . 67 , 118 . 12 , 119 . 22 , 119 . 70 , 127 . 96 , 130 . 95 , 135 . 34 , 148 . 82 , 149 . 06 , 152 . 40 , 156 . 79 , 158 . 02 ; hrms ( + esi ): m / z calcd . for ( m + h ) + , 622 . 32 . found , 623 . 33 ; uv / vis ( acetonitrile ), λ max ( ε )= 410 nm ( 21228 m − 1 cm − 1 ), λ max ( ε )= 288 nm ( 20211 m − 1 cm − 1 ). yield : 55 %, 1 h nmr ( cdcl 3 , 600 mhz ): δ = 1 . 23 ( t , 6h , j = 7 . 25 hz ), 3 . 44 ( q , 4h , j = 7 . 25 hz ,), 3 . 63 - 3 . 67 ( m , 20h ), 3 . 72 ( t , 4h , j = 5 . 68 hz ), 6 . 53 ( s , 1h ), 6 . 62 ( dd , 1h ), 6 . 76 ( d , 2h , j = 8 . 48 hz ), 7 . 41 ( d , 1h , j = 8 . 85 hz ), 7 . 76 ( d , 2h , j = 8 . 48 hz ), 8 . 60 ( s , 1h ), 8 . 64 ( s , 1h ); 13 c ( cdcl 3 , 150 mhz ): δ = 12 . 54 , 44 . 94 , 51 . 51 , 68 . 58 , 70 . 84 , 97 . 13 , 108 . 83 , 109 . 44 , 110 . 87 , 111 . 93 , 120 . 54 , 122 . 12 , 126 . 56 , 129 . 60 , 138 . 51 , 142 . 06 , 148 . 26 , 150 . 82 , 156 . 11 , 160 . 79 ; hrms ( + esi ): m / z calcd . for ( m + h ) + , 622 . 32 . found , 623 . 35 ; uv / vis ( acetonitrile ), λ max ( ε )= 413 nm ( 40057 m − 1 cm − 1 ), λ max ( ε )= 293 nm ( 18136 m − 1 cm − 1 ). the synthesis followed the general procedure of the cuaac reaction . yield : 60 %, 1 h nmr ( cdcl 3 , 300 mhz ): δ = 1 . 17 - 1 . 27 ( m , 12h ), 3 . 37 - 3 . 50 ( m , 8h ), 6 . 57 ( s , 1h ), 6 . 68 ( dd , 1h ), 6 . 75 ( d , 2h , j = 8 . 48 hz ), 7 . 42 ( d , 1h , j = 8 . 85 hz ), 7 . 76 ( d , 2h , j = 8 . 1 hz ), 8 . 44 ( s , 1h ), 8 . 65 ( s , 1h ), 13 c ( cdcl 3 , 75 mhz ): δ = 12 . 43 , 12 . 62 , 29 . 69 , 44 . 39 , 44 . 98 , 97 . 08 , 107 . 28 , 109 . 99 , 111 . 80 , 117 . 31 , 117 . 57 , 118 . 61 , 127 . 06 , 129 . 91 , 134 . 15 , 147 . 75 , 148 . 25 , 151 . 41 , 155 . 72 , 157 . 01 ; hrms ( + esi ): m / z calcd . for ( m + h ) + , 431 . 23 . found , 432 . 32 ; uv / vis ( acetonitrile ), λ max ( ε )= 410 nm ( 29501 m − 1 cm − 1 ), 248 nm ( 22823 m − 1 cm − 1 ). yield : 23 %, 1 h nmr ( cdcl 3 , 300 mhz ): δ = 1 . 14 ( t , 6h , j = 7 . 16 hz ), 2 . 70 ( m , 4h ), 3 . 36 ( q , 4h , j = 7 . 16 hz ,), 3 . 54 - 3 . 68 ( m , 20h ), 3 . 79 ( s , 2h ), 6 . 42 ( s , 1h ), 6 . 60 ( dd , 1h ), 7 . 40 ( d , 1h , j = 9 . 04 hz ), 8 . 24 ( s , 1h ), 8 . 33 ( s , 1h ); 13 c ( cdcl 3 , 75 mhz ): δ = 12 . 76 , 45 . 34 , 49 . 51 , 53 . 78 , 67 . 91 , 69 . 72 , 97 . 16 , 107 . 21 , 110 . 60 , 124 . 15 , 129 . 08 , 130 . 67 , 131 . 26 , 136 . 14 , 152 . 12 , 156 . 27 , 127 . 46 ; hrms ( + esi ): m / z calcd . for ( m + h ) + , 560 . 31 . found , 560 . 43 ; uv / vis ( acetonitrile ), λ max ( ε )= 408 nm ( 10534 m − 1 cm − 1 ), 246 nm ( 11149 m − 1 cm − 1 ). a mixture of compound 40 ( 130 mg , 0 . 37 mmol ), 3 - azido - 7 - diethylaminocoumarin 60 ( 96 . 1 mg , 0 . 37 mmol ), copper sulfate pentahydrate ( 4 . 6 mg , 5 mol %) and sodium ascorbate ( 7 . 3 mg , 10 mol %) in 6 ml thf / water ( 2 / 1 ) was stirred at 60 ° c . for 48 hours . to the reaction mixture was added water ( 5 ml ) and it was extracted 3 times with chcl 3 ( 3 × 10 ml ). the combined organic layers were dried with mgso 4 and concentrated in vacuum . the residue was purified by column chromatography on silica with chcl 3 / ch 3 oh ( 95 / 5 ) as eluent . the product was obtained as a dark yellow oil , which crystallised upon standing in a freezer . ( 90 mg , 40 %). 1 h nmr ( 500 mhz , cdcl 3 ): δ = 1 . 24 ( t , 6h , 3 j = 7 . 09 hz , 1 - h ), 3 . 45 ( q , 4h , 3 j = 7 . 09 hz , 2 - h ), 3 . 51 - 3 . 55 ( m , 4h , 21 - h ), 3 . 64 - 3 . 70 ( m , 16h , 22 - h , 23 - h , 24 - h , 25 - h ), 3 . 93 ( s , 3h , 20 - h ), 6 . 56 ( d , 1h , 4 j = 2 . 21 hz , 3 - h ), 6 . 68 ( dd , 1h , 3 j = 8 . 83 hz , 4 j = 2 . 36 hz , 5 - h ), 7 . 17 ( d , 1h , 3 j = 7 . 09 hz , 16 - h ), 7 . 38 ( d , 1h , 3 j = 7 . 88 hz , 15 - h ), 7 . 43 ( d , 1h , 3 j = 8 . 99 hz , 6 - h ), 7 . 46 ( s , 1h , 19 - h ), 8 . 44 ( s , 1h , 9 - h ), 8 . 74 ppm ( s , 1h , 12 - h ); 13 c nmr ( 125 mhz , cdcl 3 ): δ = 12 . 39 ( c1 ), 44 . 96 ( c2 ), 53 . 05 ( c21 ), 55 . 63 ( c20 ), 70 . 08 , 70 . 37 , 70 . 49 , 70 . 90 ( c22 , c23 , c24 , c25 ), 97 . 00 ( c3 ), 107 . 13 ( c7 ), 109 . 26 ( c19 ), 110 . 05 ( c5 ), 116 . 98 ( c10 ), 118 . 38 ( c15 ), 119 . 63 ( c12 ), 120 . 52 ( c16 ), 124 . 07 ( c18 ), 129 . 96 ( c6 ), 134 . 34 ( c9 ), 139 . 92 ( c13 ), 147 . 68 ( c14 ), 151 . 50 ( c4 ), 152 . 66 ( c17 ), 155 . 74 ( c8 ), 156 . 95 ppm ( c11 ) ir ( kbr , cm − 1 ): 1130 ( s ), 1239 ( s ), 1602 ( s ), 1728 ( s ), 2855 ( s ), 2923 ( s ) uv / vis ( ch 3 cn ): λ max ( ε )= 267 ( 4352 ), 413 nm ( 8649 ). a mixture of compound 43 ( 117 . 5 mg , 0 . 32 mmol ), 3 - acetylen - 7 - diethylaminocoumarin 61 ( 77 . 4 mg , 0 . 32 mmol ), cu / c ( 16 mg , 20 mol %) and triethylamin ( 32 . 5 mg , 0 . 32 mmol ) was stirred in thf ( 4 ml ) for 48 hours . cu / c was filtered off through celite , washed several times with chcl 3 and the solvent was removed under reduced pressure . the residue was purified by column chromatography on silica with chcl 3 / ch 3 oh ( 95 / 5 ) as eluent . the product was obtained as a dark yellow oil , which crystallised upon standing in a freezer . ( 91 mg , 47 %). 1 h nmr ( 300 mhz , cdcl 3 ): δ = 1 . 22 ( t , 6h , 3 j = 7 . 06 hz , 1 - h ), 3 . 43 ( q , 4h , 3 j = 7 . 06 hz , 2 - h ), 3 . 51 - 3 . 55 ( m , 4h , 21 - h ), 3 . 65 - 3 . 69 ( m , 16h , 22 - h , 23 - h , 24 - h , 25 - h ), 3 . 91 ( s , 3h , 20 - h ), 6 . 54 ( d , 1h , 4 j = 1 . 98 hz , 3 - h ), 6 . 63 ( dd , 1h , 3 j = 8 . 76 hz , 4 j = 2 . 31 hz , 5 - h ), 7 . 13 - 7 . 23 ( m , 2h , 15 - h , 16 - h ), 7 . 32 ( s , 1h , 19 - h ), 7 . 41 ( d , 1h , 3 j = 8 . 85 hz , 6 - h ), 8 . 65 ( s , 1h , 9 - h ), 8 . 66 ppm ( s , 1h , 13 - h ); 13 c nmr ( 75 mhz , cdcl 3 ): δ = 12 . 42 ( c1 ), 44 . 82 ( c2 ), 53 . 03 ( c21 ), 55 . 78 ( c20 ), 70 . 03 , 70 . 42 , 70 . 94 , ( c22 , c23 , c24 , c25 ), 97 . 03 ( c3 ), 104 . 73 ( c19 ), 108 . 67 ( c7 ), 109 . 32 ( c5 ), 110 . 59 ( c14 ), 112 . 45 ( c16 ), 120 . 40 ( c15 ), 122 . 05 ( c13 ), 129 . 49 ( c6 ), 131 . 15 ( c10 ), 138 . 49 ( c9 ), 140 . 25 ( c12 ), 142 . 14 ( c18 ), 150 . 78 ( c4 ), 152 . 83 ( c17 ), 156 . 00 ( c8 ), 160 . 62 ppm ( c11 ) ir ( kbr , cm − 1 ): 1130 ( s ), 1230 ( s ), 1600 ( s ), 1694 ( s ), 1720 ( s ), 2861 ( s ), 2969 ( s ) uv / vis ( ch 3 cn ): λ max ( ε )= 259 ( 14131 ), 410 nm ( 34604 ) elemental analysis (%) calcd . for c 32 h 41 n 5 o 7 ( 607 . 70 ): c , 63 . 25 , h , 6 . 80 , n , 11 . 52 . found c , 62 . 91 ; h , 6 . 69 ; n , 11 . 12 . the synthesis followed the general procedure of the cuaac reaction using precursors 28 and 50 . the crude product was purified by column chromatography ( 65 × 2 cm , silica gel , chcl 3 / meoh = 98 : 2 v / v ) to yield as a yellow solid . yield : 0 . 086 g , 49 %, mp : 136 . 1 - 139 . 0 ° c ., r f = 0 . 52 ; 1 h nmr , ( cdcl 3 , 600 mhz ) 6 ( ppm ), j ( hz ): 9 . 1 ( dd , 1h , h 9 ), 8 . 63 ( dd , 1h , h 7 ), 8 . 61 ( d , 1h , j = 8 . 55 , h 5 ), 8 . 26 ( s , 1h , h 10 ), 7 . 99 ( d , 1h , j = 7 . 56 , h 6 ), 7 . 79 ( dd , 1h , h 8 ), 7 . 61 , ( d , 2h , j = 9 . 12 , h 11 ), 6 . 81 ( d , 2h , j = 9 . 12 , h 12 ), 4 . 18 ( t , 2h , j = 7 . 62 , h 4 ), 3 . 74 - 3 . 64 ( m , 24h , h 13 - 18 ), 1 . 76 - 1 . 69 ( m , 2h , h 3 ), 1 . 45 ( sx , 2h , j = 7 . 62 , h 2 ), 0 . 98 ( t , 3h , j = 7 . 38 , h 1 ); 13 c nmr , ( cdcl 3 , 150 mhz ) δ ( ppm ): 164 . 34 , 164 . 06 , 148 . 68 , 146 . 15 , 145 . 66 , 134 . 32 , 132 . 92 , 131 . 52 , 130 . 81 , 129 . 40 , 128 . 95 , 127 . 48 , 127 . 35 , 125 . 96 , 122 . 94 , 122 . 56 , 121 . 52 , 120 . 06 , 112 . 96 , 111 . 99 , 70 . 98 - 70 . 82 , 68 . 8 , 68 . 60 , 51 . 59 , 40 . 41 , 30 . 32 , 20 . 52 , 13 . 99 ; hrms ( + esi ): m / z calcd for ( m + h ) + , 658 . 32 . found , 658 . 35 ; the synthesis followed the general procedure of the cuaac reaction using precursors 27 and 51 . the crude product was purified by column chromatography ( 65 × 2 cm , silica gel , chcl 3 / meoh = 95 : 5 v / v ) to yield as an orange solid . yield : 0 . 024 g , 19 %, mp : 213 . 6 - 216 . 4 ° c ., r f = 0 . 14 ; 1 h nmr , ( cdcl 3 , 600 mhz ) δ ( ppm ), j ( hz ): 8 . 71 ( t , 2h , j = 7 . 47 h 3 , h 5 ), 8 . 36 ( d , 1h , j = 8 . 1 , h 7 ), 8 . 09 ( s , 1h , h 8 ), 7 . 89 ( d , 1h , j = 7 . 47 , h a ), 7 . 83 ( tr , 1h , j = 5 . 26 , h 6 ), 7 . 79 , ( d , 2h , j = 8 . 7 , h 9 ), 6 . 79 ( d , 2h , j = 8 . 82 , h 10 ), 4 . 28 ( q , 2h , j = 7 . 12 , h 2 ), 3 . 74 - 3 . 68 ( m , 24h , h 11 - 16 ), 1 . 36 ( t , 3h , j = 7 . 12 , h 1 ); 13 c nmr , ( cdcl 3 , 150 mhz ) δ ( ppm ): 163 . 68 , 163 . 16 , 149 . 13 , 148 . 48 , 138 . 59 , 132 . 29 , 130 . 82 , 129 . 87 , 129 . 29 , 128 . 63 , 127 . 31 , 126 . 71 , 123 . 93 , 123 . 46 , 123 . 19 , 120 . 20 , 117 . 23 , 112 . 01 , 71 . 04 - 70 . 93 , 68 . 82 , 51 . 50 , 35 . 93 , 13 . 47 ; hrms ( + esi ): m / z calcd for ( m + h ) + , 630 . 29 . found , 630 . 14 the synthesis followed the general procedure of the cuaac reaction using precursors 28 and 52 . the crude product was purified by column chromatography ( 65 × 2 cm , silica gel , chcl 3 / meoh = 95 : 5 v / v ) to yield as a light brown oil . yield : 23 %, 1 h nmr ( cdcl 3 , 300 mhz ): δ = 3 . 53 - 3 . 758 ( m , 24h , h 914 ), 6 . 31 ( d , 2h , j = 9 . 231 , h 8 ), 7 . 40 - 7 . 55 ( m , 4h , h 4 , 2 ), 7 . 7 ( d , 2h , j = 9 . 231 , h 7 ), 8 . 03 ( m , 4h , h 5 , 2 ), 8 . 1 ( s , 1h , h 6 ), 8 . 57 ( s , 1h , h 1 ); 13 c ( cdcl 3 , 75 mhz ): δ = 51 . 16 , 68 . 37 , 70 . 85 , 111 . 24 , 119 . 95 , 122 . 07 , 124 . 54 , 125 . 21 , 125 . 48 , 126 . 07 , 126 . 89 , 128 . 64 , 131 . 02 , 131 . 34 , 145 . 62 , 148 . 02 ; hrms ( + esi ): m / z calcd . ( m + h ) + , 583 . 34 . found , 583 . 29 ; uv / vis ( mecn ,) λ max ( ε )= 388 nm ( 5596 m − 1 cm − 1 ), 420 nm ( 5782 m − 1 cm − 1 ). the synthesis followed the general procedure of the cuaac reaction using precursors 56 and 61 . the crude product was purified by column chromatography ( 65 × 2 cm , silica gel . chcl 3 / meoh = 95 : 5 v / v ) to yield as a yellow oil . yield : 13 %, hrms ( + esi ): m / z calc . ( m + h ) + . 696 . 36 . found . 696 . 47 . the synthesis followed the general procedure of the cuaac reaction using precursors 56 and 52 . the crude product was purified by column chromatography ( 65 × 2 cm , silica gel . chcl 3 / meoh = 98 : 2 v / v ) to yield 5 as a light brown oil . yield : 19 . 3 %; 1 h nmr ( cdcl 3 . 600 mhz ): δ = 3 . 35 - 3 . 75 ( m . 27h ). 3 . 81 ( tr . 2h ). 4 . 28 ( tr . 2h ). 7 . 31 ( d . 1h ). 7 . 45 ( m . 5h ). 7 . 58 ( s . 1h ). 7 . 91 ( d . 2h ). 8 . 05 ( d . 2h ). 8 . 21 ( s . 1h ). 8 . 55 ( s . 1h ); the synthesis followed the general procedure of the cuaac reaction using precursors 56 and 57 . the crude product was purified by column chromatography ( 65 × 2 cm , silica gel . chcl 3 / meoh = 95 : 5 v / v ) to yield 9 as a light brown oil . yield : 24 %; 1 h nmr ( cdcl 3 600 mhz ): δ = 0 . 9 ( tr . 3h ). 1 . 27 ( m . 24h ). 1 . 39 ( qu . 2h ). 1 . 47 ( qu . 2h ). 1 . 78 ( qu . 2h ). 3 . 40 - 3 . 78 ( m . 27h ). 3 . 83 ( tr . 2h ). 4 . 28 ( tr . 2h ). 7 . 21 ( d . 1h ). 7 . 35 ( d . 1h ). 7 . 50 ( s . 1h ). 7 . 86 ( tr . 1h ). 8 . 09 ( d . 1h ). 8 . 49 ( s . 1h ). 8 . 7 ( tr . 2h ). 9 . 17 ( d . 1h ). the synthesis followed the general procedure of the cuaac . yield : 62 %, hrms ( + esi ): m / z calc . 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