Patent Application: US-63254505-A

Abstract:
the present invention relates to compounds with α7 nachr agonistic activity , processes for their preparation , pharmaceutical compositions containing the same and the use thereof for the treatment of neurological , psychiatric , cognitive , immunological and inflammatory disorders .

Description:
in a first aspect , the invention provides a compound of formula i y is a group — conh —; — nhconh —; — nhco —; — so 2 nh —; — nhso 2 —; — nhso 2 nh —; — oconh ; — nhcoo — r is hydrogen ; halogen ; linear , branched or cyclic ( c 1 - c 6 ) alkyl , haloalkyl , alkoxy or acyl ; hydroxy ; cyano ; nitro ; mono - or di - ( c 1 - c 6 ) alkylamino , acylamino or alkylaminocarbonyl ; carbamoyl ; ( c 6 - c 10 ) aryl - or ( c 1 - c 6 ) alkylsulphonylamino ; ( c 6 - c 10 ) aryl - or ( c 1 - c 6 ) alkylsulphamoyl ; a 5 to 10 - membered aromatic or heteroaromatic ring optionally substituted with : halogen ; linear , branched or cyclic ( c 1 - c 3 ) alkyl , haloalkyl , alkoxy or acyl ; hydroxy ; cyano ; nitro ; amino ; mono - or di - ( c 1 - c 6 ) alkylamino , acylamino or alkylaminocarbonyl groups ; carbamoyl ; ( c 6 - c 10 ) aryl - or ( c 1 - c 6 ) alkylsulphonylamino ; ( c 6 - 10 ) aryl - or ( c 1 - c 6 ) alkylsulphamoyl ; r 1 represents ( c 1 - c 6 ) acyl ; linear , branched or cyclic ( c 1 - c 6 ) alkyl ; a —( ch 2 ) j — r ′″ group , wherein j = 0 , 1 and r ′″ is a 5 to 10 - membered aromatic or heteroaromatic ring optionally substituted with : halogen ; hydroxy ; cyano ; nitro ; ( c 1 - c 6 ) alkyl , haloalkyl , alkoxy , acyl , acylamino groups ; r ″, independently from one another for p = 2 , represents hydrogen ; halogen ; hydroxy ; cyano ; nitro ; linear , branched or cyclic ( c 1 - c 6 ) alkyl , haloalkyl , alkoxy , acyl ; a —( ch 2 ) j — r ′″ group , wherein n and r ′″ are as above defined ; carbamoyl ; ( c 6 - c 10 ) aryl - or ( c 1 - c 3 ) alkylsulphonylamino ; ( c 6 - c 10 ) aryl - or ( c 1 - c 3 ) alkylsulphamoyl ; mono - or di -[ linear , branched or cyclic ( c 1 - c 6 ) alkyl ] aminocarbonyl ; a first group ( ia ) of preferred compounds of formula i are those in which : r is selected from the group consisting of hydrogen ; halogen ; linear , branched or cyclic ( c 1 - c 6 ) alkyl , alkoxy or alkylamino ; trihaloalkyl ; phenyl ; naphthyl ; pyridyl ; pyrimidinyl ; quinolinyl ; isoquinolinyl ; indolyl ; thienyl ; benzothienyl ; furanyl ; benzofuranyl ; imidazolyl ; benzoimidazolyl ; pyrrolyl ; optionally substituted as indicated above for the compounds of formula ( i ); r ″, independently from one another for p = 2 , is selected from the group consisting of hydrogen ; mono - or di -[ linear , branched or cyclic ( c 1 - c 6 ) alkyl ] aminocarbonyl ; linear , branched or cyclic ( c 1 - c 6 ) alkyl , alkoxy , acyl ; r is selected from the group consisting of phenyl ; naphthyl ; pyridyl ; pyrimidinyl ; quinolinyl ; isoquinolinyl ; indolyl ; thienyl ; benzothienyl ; furanyl ; benzofuranyl ; imidazolyl ; benzoimidazolyl ; pyrrolyl ; optionally substituted as indicated above for the compounds of formula ( i ); r ″, independently of one another for p = 2 , is selected from the group consisting of hydrogen ; mono - or di -[ linear , branched or cyclic ( c 1 - c 6 ) alkyl ] aminocarbonyl ; linear , branched or cyclic ( c 1 - c 6 ) alkyl , alkoxy , acyl ; r is selected from the group consisting of halogen ; linear , branched or cyclic ( c 1 - c 6 ) alkyl , alkoxy or alkylamino ; haloalkyl ; phenyl ; naphthyl ; pyridyl ; pyrimidinyl ; quinolinyl ; isoquinolinyl ; indolyl ; thienyl ; benzothienyl ; furanyl ; benzofuranyl ; imidazolyl ; benzoimidazolyl ; pyrrolyl ; optionally substituted as indicated above for the compounds of formula ( i ); r ″, independently from one another for p = 2 , is selected from the group consisting of hydrogen ; mono - or di -[ linear , branched or cyclic ( c 1 - c 6 ) alkyl ] aminocarbonyl ; linear , branched or cyclic ( c 1 - c 6 ) alkyl , alkoxy , acyl ; r is selected from the group consisting of phenyl ; naphthyl ; pyridyl ; pyrimidinyl ; quinolinyl ; isoquinolinyl ; indolyl ; thienyl ; benzothienyl ; furanyl ; benzofuranyl ; imidazolyl ; benzoimidazolyl ; pyrrolyl ; optionally substituted as indicated above for the compounds of formula ( i ); r ″, independently of one another for p = 2 , is selected from the group consisting of hydrogen ; mono - or di -[ linear , branched or cyclic ( c 1 - c 6 ) alkyl ] aminocarbonyl ; linear , branched or cyclic ( c 1 - c 6 ) alkyl , alkoxy , acyl ; a further group ( ib ) of preferred compounds of formula ( i ) are those in which r is selected from the group consisting of halogen ; phenyl ; naphthyl ; pyridyl ; quinolinyl ; isoquinolinyl ; indolyl ; thienyl ; benzothienyl ; furanyl ; benzofuranyl ; imidazolyl ; benzoimidazolyl ; pyrrolyl ; optionally substituted as indicated above for the compounds of formula ( i ); where r ′ is a 5 - 10 - membered aromatic or heteroaromatic ring optionally substituted with halogen or ( c 1 - c 6 ) alkoxy groups ; a further group ( ic ) of preferred compounds of formula ( i ) are those in which r is selected from the group consisting of halogen ; phenyl ; naphthyl ; pyridyl ; quinolinyl ; isoquinolinyl ; indolyl ; thienyl ; benzothienyl ; furanyl ; benzofuranyl ; imidazolyl ; benzoimidazolyl ; pyrrolyl ; optionally substituted as indicated above for the compounds of formula ( i ); where r ′ is a 6 - membered aromatic or heteroaromatic ring optionally substituted with halogen or ( c 1 - c 6 ) alkoxy groups ; another group ( id ) of preferred compounds of formula i are those in which r is selected from the group consisting of phenyl ; naphthyl ; pyridyl ; quinolinyl ; pyrimidinyl ; isoquinolinyl ; indolyl ; thienyl ; benzothienyl ; furanyl ; benzofuranyl ; imidazolyl ; benzoimidazolyl ; pyrrolyl ; optionally substituted as indicated above for the compounds of formula ( i ); where r ′ is a phenyl ring optionally substituted with halogen or ( c 1 - c 6 ) alkoxy groups ; r is selected from the group consisting of phenyl ; pyridyl ; indolyl ; pyrimidinyl ; optionally substituted with : halogen ; linear , branched or cyclic ( c 1 - c 3 ) alkyl , alkoxy or acyl ; cyano ; ( c 1 - c 6 ) alkylamino ; acylamino ; alkylaminocarbonyl groups ; carbamoyl ; where r ′ is a phenyl ring optionally substituted with halogen or ( c 1 - c 6 ) alkoxy groups the compounds of the invention can be in the form of free bases or acid addition salts , preferably salts with pharmaceutically acceptable acids . the invention also includes separated isomers and diastereomers of compounds i , or mixtures thereof ( e . g . racemic mixtures ). the compounds of formula ( i ) can be prepared through a number of synthetic routes amongst which the ones illustrated in schemes 1 , 2 , and 3 ( see also for reference bioorg . med . chem . lett . 1995 , 5 ( 3 ), 219 - 222 ). according to scheme 1 , a suitably activated butylphthalimide ( compound 2 ) is reacted with an amine ( compound 1 ) in an organic solvent in the presence of a base . for example , a mixture of 1 ( or its hydrochloride salt ) and 2 are refluxed in methylethyl ketone in the presence of alkaline carbonate until the reaction is complete , then the reaction mixture is cooled , the insoluble materials removed by filtration , the filtrate washed with chcl 3 , and the filtrate and washings concentrated to dryness . in the following step , the n -( 4 - aminobutyl ) phthalimide 3 is converted into a ( 4 - aminobutyl ) amine 4 , for example by refluxing a mixture of 3 and hydrazine hydrate in ethanol . then 4 is reacted with an activated species 5 such as for example ( but not limited to ) an acid chloride or an isocyanate in an organic solvent in the presence of a base . for example , to a mixture of 4 and 5 in ch 2 cl 2 triethylamine and a catalytic amount of dmap are added , to give compounds i . alternatively , a mixture of 4 , 5 , a carbodiimide or carbonyldiimidazole and dmap are reacted to yield compounds i . according to scheme 2 , aminobutanol is reacted with an activated acid species or an isocyanate — for example ( but not limited to ) a substituted acid chloride 6 in the presence of a base — in an organic solvent like dichloromethane until the reaction is complete . the alcohol 7 thus obtained is then oxidised under standard conditions ( for example swern oxidation ) and aldehyde 8 is then reacted with the suitably substituted amine 1 under standard conditions — for example with sodium triacetoxyborohydride — to afford compound iα . in the case of r being a halogen , iα can be further processed — for example via a cross - coupling reaction with a boronic acid — to yield compound iβ . according to scheme 3 , 5 - bromopentanoyl chloride is reacted with an ( hetero ) aromatic amine 9 in the presence of an organic base to afford a 5 - bromopentanoic acid amide 10 . this species is reacted with an amine 1 to displace the halogen and furnish compounds iα . in the case of r being a halogen , ia can be further processed — for example via a cross - coupling reaction with a boronic acid — to yield compounds iβ . the compounds of formula i , their optical isomers or diastereomers can be purified or separated according to well - known procedures , including but not limited to chromatography with chiral matrix and fractional crystallisation . the pharmacological activity of a representative group of compounds of formula i was demonstrated in an in vitro assay utilising cells stably transfected with the alpha 7 nicotinic acetylcholine receptor and cells expressing the alpha 1 and alpha 3 nicotinic acetylcholine receptors and 5ht3 receptor as controls for selectivity . neuroprotection of these compounds was demonstrated in a cell - based excitotoxicity assay utilising primary neuronal cell cultures . according to a further aspect , the invention is therefore directed to a method of treating neurological and psychiatric disorders , which comprises administering to a subject , preferably a human subject in need thereof , an effective amount of a compound of formula i . neurological and psychiatric disorders that may benefit from the treatment with the invention compounds include but are not limited to senile dementia , attention deficit disorders , alzheimer &# 39 ; s disease and schizophrenia . in general , the compounds of formula i can be used for treating any disease condition , disorder or dysfunction that may benefit from the activation of the alpha 7 nicotinic acetylcholine receptor , including but not limited to parkinson &# 39 ; s disease , huntington &# 39 ; s chorea , amyotrophic lateral sclerosis , multiple sclerosis , epilepsy , memory or learning deficit , panic disorders , cognitive disorders , depression , sepsis , arthritis , immunological and inflammatory disorders . the dosage of the compounds for use in therapy may vary depending upon , for example , the administration route , the nature and severity of the disease . in general , an acceptable pharmacological effect in humans may be obtained with daily dosages ranging from 0 . 01 to 200 mg / kg . in yet a further aspect , the invention refers to a pharmaceutical composition containing one or more compounds of formula i , in association with pharmaceutically acceptable carriers and excipients . the pharmaceutical compositions can be in the form of solid , semi - solid or liquid preparations , preferably in form of solutions , suspensions , powders , granules , tablets , capsules , syrups , suppositories , aerosols or controlled delivery systems . the compositions can be administered by a variety of routes , including oral , transdermal , subcutaneous , intravenous , intramuscular , rectal and intranasal , and are preferably formulated in unit dosage form , each dosage containing from about 1 to about 1000 mg , preferably from 1 to 600 mg of the active ingredient . the compounds of the invention can be in the form of free bases or as acid addition salts , preferably salts with pharmaceutically acceptable acids . the invention also includes separated isomers and diastereomers of compounds i , or mixtures thereof ( e . g . racemic mixtures ). the principles and methods for the preparation of pharmaceutical compositions are described for example in remington &# 39 ; s pharmaceutical science , mack publishing company , easton ( pa .). effect of compound from example 64 on nmda - induced toxicity in rat cortical neurons . rat cortical neurons were pre - treated with the compound at the indicated concentrations 24 h before addition of nmda and toxicity determined by lactate dehydrogenase ( ldh ) measurements after 24 h . data of all experiments are normalised to 100 % nmda toxicity . statistical analysis : p & lt ; 0 . 05 vs nmda treatment ; one - way anova and tukey post test values were normalised to the level of nmda (= 100 %). effect of sub - chronic treatment of compound from example 1 or nicotine on number of chat - positive neurons in the nucleus basalis of quisqualic acid injected animals . compounds were administered 24 h and 1 h before quisqualic acid injection and for 7 days after lesioning . doses : compound 3 mg / kg i . p . daily or nicotine 0 . 3 mg / kg i . p . daily . the doses were selected on the basis of literature data and comparable effects in behavioral studies . number of neurons is expressed as % changes vs non - injected hemisphere . statistical analysis : anova and fisher post - hoc test : f ( 3 , 21 )= 13 . 00 p & lt ; 0 . 001 * p & lt ; 0 . 05 vs quisqualic acid injected rats # p & lt ; 0 . 05 vs nicotine treated rats . effect of acute administration of compound from example 1 on scopolamine - induced amnesia in young rats in passive avoidance test and reversion by the selective alpha - 7 antagonist mla . amnesia was induced by scopolamine 0 . 5 mg / kg i . p . 20 min before training trial and the compound ( 3 mg / kg i . p .) was injected 5 min after scopolamine . mla ( 5 mg / kg i . p .) was administered 10 min before scopolamine and compound administration . results are presented as retest latencies 24 h after the training trial . statistical analysis : anova and tukey post - hoc test : * p & lt ; 0 . 05 vs saline and scopolamine - treated rats # p & lt ; 0 . 05 vs saline treated rats . effect of acute administration of compound from example 1 on scopolamine - induced amnesia in young rats . amnesia was induced by scopolamine 0 . 2 mg / kg i . p . 20 min before training trial and the compound ( 3 mg / kg i . p .) was injected 5 min after scopolamine . results are presented as discrimination index calculated on the exploration time of new ( n ) and familiar ( f ) objects during the test trial performed after 2 h from the training trial as follow : discrimination index : n − f / n + f . statistical analysis : anova and tukey post - hoc test : * p & lt ; 0 . 05 scopolamine - treated rats . unless otherwise specified all nuclear magnetic resonance spectra were recorded using a bruker ac200 ( 200 mhz ) or a varian mercury plus 400 mhzspectrometer equipped with a pfg atb broadband probe . hplc - ms analyses were performed with an agilent 1100 instrument , using a zorbax eclipse xdb - c8 4 . 6 × 150 mm ; a zorbax cn 4 . 6 × 150 mm column or a zorbax extend c18 2 . 1 × 50 mm column , coupled to an atmospheric api - es ms for the 2 . 5 minutes method . the 5 and 10 minute methods were run using a waters 2795 separation module equipped with a waters micromass zq ( es ionisation ) and waters pda 2996 , using a waters xterra ms c18 3 . 5 μm 2 . 1 × 50 mm column . preparative hlpc was run using a waters 2767 system with a binary gradient module waters 2525 pump and coupled to a waters micromass zq ( es ) or waters 2487 dad , using a supelco discovery hs c18 5 . 0 μm 10 × 21 . 2 mm column gradients were run using 0 . 1 % formic acid / water and 0 . 1 % formic acid / acetonitrile with gradient 5 / 95 to 95 / 5 in the run time indicated . all column chromatography was performed following the method of still , c . ; j . org chem 43 , 2923 ( 1978 ). all tlc analyses were performed on silica gel ( merck 60 f254 ) and spots revealed by uv visualisation at 254 nm and kmno4 or ninhydrin stain . the compounds were prepared following the general procedure outlined in nishikawa , y . ; et al ; chem . pharm . bull ., 1989 , 37 ( 1 ), 100 - 105 . a mixture of n -( 4 - bromobutyl )- phthalimide ( 0 . 00135 mol ), 1 -( aryl )- piperazine hydrochloride ( 0 . 00135 mol ), k 2 co 3 ( 0 . 00270 mol ), nai ( 0 . 00186 mol ) and methylethyl ketone ( 7 ml ) was refluxed for 20 h with stirring . after the mixture was cooled , the insoluble materials were removed by filtration and washed with chcl 3 . the filtrate and the washings were concentrated to dryness in vacuo . the residue was subjected to chromatography on silica gel using chcl 3 / meoh 95 / 5 as eluent . a solution of n -( 4 -( arylpiperazin - 1 - yl )- butyl ) phthalimides ( 0 . 236 mmol ) and hydrazine hydrate ( 0 . 478 mmol ) in ethanol ( 2 ml ) was refluxed for 2 h with stirring . after the solution had cooled , the insoluble materials were removed by filtration and washed with etoh . the filtrate and the washings were concentrated to dryness in vacuo . the residue was taken up with chcl 3 . the chcl 3 layer was washed with water , dried and concentrated to give the title amine . a ) following the general procedure , 2 - methoxyphenyl - piperazine ( 3 . 4 ml , 17 . 7 mmol ) is added to a suspension of n -( 4 - bromobutyl ) phthalimide ( 5 g , 17 . 7 mmol ), sodium iodide ( 1 . 33 g , 8 . 85 mmol ) and potassium carbonate ( 3 . 67 g , 26 . 6 mmol ) in 2 - butanone ( 70 ml ). the resulting suspension is stirred for 18 h at 100 ° c ., before lc - ms check . the reaction is filtered and the solvent removed by vacuum distillation ; the resulting oil is dissolved in 5 % meoh in dichloromethane , washed with water and sat . nacl , dried over na 2 so 4 . the solvent is removed under reduced pressure to yield the desired product as a thick yellow oil . the residue is extracted into ethyl acetate and washed with water and then saturated brine and dried over sodium sulphate . the solvent is removed under reduced pressure to afford 5 . 01 g of 2 -{ 4 -[ 4 -( 2 - methoxy - phenyl )- piperazin - 1 - yl ]- butyl }- isoindole - 1 , 3 - dione used without further purification in step b ) below ( 72 %). 2 -{ 4 -[ 4 -( 2 - methoxy - phenyl )- piperazin - 1 - yl ]- butyl }- isoindole - 1 , 3 - dione ( 5 . 01 g , 12 . 7 mmol ) is dissolved in abs . etoh ( 60 ml ) and hydrazine monohydrate ( 2 . 54 ml , 26 mmol ) is added dropwise . the reaction is heated at 100 ° c . for 1 h ; the reaction is filtered , concentrated at reduced pressure and transformed into its hydrochloride salt . the salt is dissolved in 15 % naoh and extracted into ethyl acetate to yield 2 . 04 g of 4 -[ 4 -( 2 - methoxy - phenyl )- piperazin - 1 - yl ]- butylamine as waxy solid ( 7 . 8 mmol , 61 %). c 15 h 25 n 3 o mass ( calculated ) [ 263 . 39 ]; ( found ) [ m + h +] 32 264 . 39 nmr ( 400 mhz , cdcl3 ): 1 . 48 ( 2h , m ); 1 . 57 ( 2h , m ); 2 . 42 ( 2h , m ); 2 . 65 ( 4h , bs ); 2 . 72 ( 2h , m ); 3 . 1 ( 4h , bs ); 3 . 86 ( 3h , s ); 6 . 85 ( 1h , d ); 6 . 97 ( 3h , m ). to a solution of n -( 4 - bromobutyl ) phthalimide ( 5 g , 17 . 73 mmol ) and 1 -( 2 , 4 - difluoro - phenyl )- piperazine ( 17 . 73 mmol ) in 2 - butanone ( 100 ml ), potassium carbonate ( 26 . 6 mmol ) and potassium iodide ( 13 . 3 mmol ) were added . the resulting mixture was heated at 90 ° c . overnight . after cooling the solution was filtered and evaporated to dryness . the residue was dissolved in dichloromethane ( 100 ml ) and washed with water . the organic phase was dried over sodium sulphate and evaporated . this material was dissolved in ethanol ( 100 ml ) and hydrazine ( 2 eq ) was added . the solution was refluxed for 4 hours when a thick precipitate formed . conc . hcl ( 5 ml ) was then added and the mixture heated for a further hour . after cooling the solvent was evaporated and the residue dissolved in 2m hcl ( 100 ml ). this solution was filtered and the aqueous filtrate evaporated again to dryness . the resulting residue was taken in isopropanol ( 30 ml ) and filtered to give the hydrochloride salt of the required product . the salt was converted in the free amine by dissolution in naoh ( 15 % w / w ) and extraction with dichloromethane . ( 2 . 6 g , 54 %). 1 h - nmr ( cdcl 3 ) δ 1 . 3 ( br s , 2h ), 1 . 46 - 1 . 58 ( m , 4h ), 2 . 41 ( t , 2h ), 2 . 62 ( s , 4h ), 2 . 73 ( t , 2h ), 3 . 05 ( br s , 4h ), 6 . 77 - 6 . 83 ( m , 2h ), 6 . 87 - 6 . 94 ( m , 1h ) ( m + 1 ) e / z 270 a ) following the general procedure , morpholine ( 1 . 7 ml , 20 mmol ) is added to a suspension of n -( 4 - bromobutyl ) phthalimide ( 5 . 36 g , 20 mmol ), sodium iodide ( 1 . 5 g , 10 mmol ) and potassium carbonate ( 5 . 53 g , 40 mmol ) in 2 - butanone ( 80 ml ). the resulting suspension is stirred for 18 h at 100 ° c ., before lc - ms check . the reaction is filtered and the solvent removed by vacuum distillation ; the resulting oil is dissolved in 5 % meoh in dichloromethane , washed with water and sat . nacl , dried over na 2 so 4 . the solvent is removed under reduced pressure to yield the desired product as a thick yellow oil . the residue was extracted into ethyl acetate and washed with water and then saturated brine and dried over sodium sulphate . the solvent was removed under reduced pressure to afford 5 . 7 g of 2 -( 4 - morpholin - 4 - yl - butyl )- isoindole - 1 , 3 - dione used without further purification in step b ) below . c 16 h 20 n 2 o 3 mass ( calculated ) [ 288 . 35 ]; ( found ) [ m + h +]= 289 . 36 b ) 4 - morpholin - 4 - yl - butyl - isoindole - 1 , 3 - dione ( 5 . 69 g , 19 mmol ) is dissolved in abs . etoh ( 95 ml ) and hydrazine monohydrate ( 3 . 8 ml , 80 mmol ) is added dropwise . the reaction is heated at 100 ° c . for 1 h ; lc - ms show the reaction to be complete . the reaction is filtered , concentrated at reduced pressure and taken up with toluene and dichloromethane to remove excess phthalhydrazide ; the crude amine is purified by scx column , eluting with meoh : dichloromethane 1 : 1 followed by 2 m nh3 in meoh , to afford 1 . 46 g ( 9 . 2 mmol , 48 %). c 8 h 18 n 2 o mass ( calculated ) [ 158 . 25 ]; ( found ) [ m + h +]= 159 . 27 nmr ( 400 mhz , cd3od ): 1 . 51 ( 4h , m ); 2 . 36 ( 2h , m ); 2 . 46 ( 4h , s ); 2 . 64 ( 2h , m ); 3 . 68 ( 4h , m ). 1 h - nmr ( cdcl 3 ) δ 1 . 26 ( br s , 2h ), 1 . 44 - 1 . 57 ( m , 4h ), 2 . 35 ( t , 2h ), 2 . 44 ( br s , 4h ), 2 . 71 ( t , 2h ), 3 . 72 ( m , 4h ) 1 h - nmr ( dmso - d6 + d 2 o ) δ 1 . 53 - 1 . 61 ( m , 2h ), 1 . 66 - 1 . 74 ( m , 2h ), 2 . 80 ( t , 2h ), 2 . 85 ( s , 3h ), 3 . 17 ( m , 2h ), 3 . 38 ( br s , 4h ), 3 . 67 ( br s , 4h ); ( m + 1 ) e / z 172 . a ) following the general procedure , n -( 4 - bromobutyl ) phthalimide ( 5 . 96 g , 20 mmol ) was added to a suspension of piperidine ( 1 . 98 ml , 20 mmol ), sodium iodide ( 1 . 5 g , 10 mmol ) and potassium carbonate ( 4 . 15 g , 21 mmol ) in 2 - butanone ( 100 ml ). the resulting suspension was stirred for 18 h at 85 ° c . the reaction was filtered and the solvent removed by vacuum distillation ; the resulting oil was washed with water and recovered with dichloromethane . the solvent was removed under reduced pressure to afford 3 . 7 g of desired product as a white solid ( yield : 65 %). c 17 h 22 n 2 o 2 mass ( calculated ) [ 286 . 38 ]; ( found ) [ m + h +]= 287 nmr ( 400 mhz , cdcl3 ) 1 . 41 ( 2h , m ), 1 . 49 - 1 . 59 ( 6h , m ), 1 . 65 - 1 . 72 ( 2h , m ), 2 . 15 - 2 . 35 ( 6h , m ), 3 . 69 - 3 . 73 ( 6h , m ), 7 . 69 - 7 . 74 ( 2h , m ), 7 . 80 - 7 . 85 ( 2h , m ). b ) 2 -( 4 - piperidin - 1 - yl - butyl )- isoindole - 1 , 3 - dione ( 3 . 7 g , 13 mmol ) was dissolved in etoh ( 50 ml ) and hydrazine monohydrate ( 1 . 26 ml , 26 mmol ) was added dropwise . the mixture was heated at 80 ° c . for 4 h . the reaction was filtered , concentrated at reduced pressure and taken up with toluene and dichloromethane to remove excess phthalhydrazide by filtration ; the crude amine was purified by scx column , eluting with meoh : dichloromethane 1 : 1 followed by 2 m nh3 in meoh , to afford g ( 410 mg , 35 %). c 9 h 20 n 2 mass ( calculated ) [ 156 . 27 ]; ( found ) [ m + h +] 157 nmr ( 400 mhz , cd3od ): 1 . 45 - 1 . 62 ( 10 h , m ), 2 . 30 - 2 . 43 ( 10 h , m ), 2 . 64 - 2 . 67 ( 2h , m ). a ) following the general procedure , commercially available n , n - diethylnipecotamide ( 3 . 4 g , 40 mmol ) was weighed , placed in a flask and dissolved in 150 ml 2 - butanone . to this n -( 4 - bromobutyl ) phthalimide ( 11 . 3 g , 40 mmol ), nai ( 3 g , 20 mmol ) and k2co3 ( 8 . 28 g , 60 mmol ) were added . the resulting mixture was heated at 85 ° c . for 20 hours . the solution was dried under vacuum and the crude solution was washed twice with water and dichloromethane . the organic layer was purified by flash chromatography using dichloromethane / meoh 96 / 4 . c 22 h 31 n 3 o 3 mass ( calculated ) [ 385 . 50 ]; ( found ) [ m + h +]= 386 nmr ( 400 mhz , cdcl3 ): 1 . 08 - 1 . 12 ( 2h , m ), 1 . 14 - 1 . 21 ( 2h , m ), 1 . 52 - 1 . 76 ( 8h , m ), 2 . 1 ( 1h , m ), 2 . 23 ( 1 h , m ), 2 . 44 ( 1h , m ), 2 . 79 ( 1h , m ), 2 . 94 ( 2h , m ), 3 . 29 - 3 . 35 ( 4h , m ), 3 . 69 - 3 . 73 ( 2h , m ), 7 . 71 - 7 . 82 ( 2h , m ), 7 . 82 - 7 . 86 ( 2h , m ). b ) the phthalimide was deprotected using the general method described for the previous examples to obtain the desired product in 38 % yield . c 14 h 29 n 3 o mass ( calculated ) [ 255 . 23 ]; ( found ) [ m + h +] 32 256 nmr ( 400 mhz , cdcl3 ): 1 . 09 ( 3h , m ); 1 . 21 ( 3h , m ); 1 . 50 - 1 . 60 ( 1h , m ); 1 . 62 - 1 . 84 ( 6h , m ), 2 . 13 - 2 . 19 ( 1h , m ); 2 . 35 - 2 . 40 ( 1h , m ); 2 . 46 - 2 . 50 ( 2h , m ); 2 . 79 - 3 . 02 ( 5h , m ); 3 . 27 - 3 . 47 ( 4h , m ); 5 . 20 - 5 . 31 ( 3h , m ). prepared according to the procedure outlined in gong , y . and pauls , h . w . synlett , 2000 , 6 , 829 - 831 . a catalytic amount of pd ( pph 3 ) 4 was added to a degassed solution of 4 - carboxyphenylboronic acid ( 0 . 001 mol ) and arylic bromide ( 0 . 001 mol ) in 0 . 4 m sodium carbonate solution ( 5 ml ) and acetonitrile ( 5 ml ). the mixture was heated at 90 ° c . under n 2 for 15 - 20 h . the hot suspension was filtered . the filtrate was concentrated to about a half the original volume and then washed with ch 2 cl 2 . the aqueous layer was acidified with conc . hcl and the resulting precipitate was collected . 1 h - nmr ( cd 3 od ) δ ( ppm ): 8 . 10 ( d , 1h ); 7 . 50 ( d , 2h ); 6 . 94 ( m , 4h ) 1 h - nmr ( cd 3 od ) δ ( ppm ): 8 . 63 ( d , 1h ); 8 . 05 ( m , 4h ); 7 . 90 ( m , 2h ); 7 . 51 ( m , 1h ). prepared with a modification of the procedure outlined in leadbeater , n . e . ; marco , m ; org . lett . 2002 , 4 917 ) 2973 - 2976 : in a 10 ml glass tube were placed 4 - carboxyphenyl boronic acid ( 166 mg , 1 . 0 mmol ), 2 - bromotoluene ( 120 μl , 1 . 0 mmol ), na 2 co 3 ( 315 mg , 3 mmol ), pd ( oac ) 2 ( 1 mg , 0 . 004 mmol ), 2 ml of water and a magnetic stirbar . the vessel was sealed with a septum and placed into the microwave cavity . microwave irradiation ( maximum emitted power 200w ) was used to increase the temperature to 150 ° c . ; the reaction mixture was then kept at this temperature for 5 min . the mixture was allowed to cool to room temperature , and the reaction mixture was filtered washing with little chcl 3 . the aqueous layer was acidified , and the precipitate collected . the product was purified by chromatography on silica gel using petroleum ether / acoet 50 / 50 as eluent to give 67 . 8 mg of 12 , yield 32 %. 1 h - nmr ( cd 3 od ) δ ( ppm ): 8 . 05 ( m , 2h , arom ); 7 , 41 ( m , 2h , arom ); 7 . 21 ( m , 4h , arom ); 2 . 22 ( s , 3h , c — ch 3 ). to a stirred solution of 2 ′- aminobiphenyl - 4 - carboxylic acid ( 213 mg , 0 . 001 mol ) in hexane / water / acetone ( 6 . 7 : 5 : 1 , 6 ml ), were added at 0 ° c . nahco 3 ( 400 mg ) and oxone ( 1 . 050 g ). after 20 min a second portion of nahco 3 ( 400 mg ) and oxone ® ( 1050 mg ) was added and , after 20 min , a final portion of nahco 3 ( 400 mg ) and oxone ® ( 1050 mg ) was added . after 6 h the suspension was diluted with water and the organic layer was extracted with ch 2 cl 2 . the combined organic layers were evaporated to give 2 ′- nitro - biphenyl - 4 - carboxylic acid ( 138 . 5 mg , 0 . 00057 mol ), yield 57 %. mass ( es neg ) m / z %: 242 ( m − 1 , 100 %); 226 ( m − 1 - 16 , 70 %) to a solution of 4 - carboxyphenylboronic acid ( 3 . 32 g , 20 mmol ), fibrecat ® 1007 ( 2 g ) and potassium carbonate ( 3 . 03 g , 22 mmol ) in ethanol / water ( 20 ml / 20 ml ), 1 - bromo - 2 - methoxy - benzene was added ( 4 . 11 g , 22 mmol ). the reaction mixture was heated to reflux for 3 hours . after cooling , was filtered and the solution evaporated under reduced pressure . the residue was suspended in aq . citric acid ( 10 % w / v ), filtered and washed with water and diethyl ether . the resulting solid was dried under vacuum to yield the title compound ( 4 . 02 g , 88 %). 1 h - nmr ( dmso - d6 ) δ 3 . 79 ( s , 3h ), 7 . 08 ( m , 1h ), 7 . 34 ( m , 1h ), 7 . 58 ( d , 1h ), 7 . 96 ( d , 1h ) a mixture of 4 - carboxyphenylboronic acid ( 3 . 32 g , 20 mmol ), fibrecat ® 1007 ( 1 g ), potassium carbonate ( 3 . 03 g , 22 mmol ) and 1 - bromo - 2 - chloro - benzene ( 4 . 2 g , 22 mmol ) were exposed to microwave irradiation in a cem discovery microwave for 15 minutes up to the maximum temperature of 120 ° c . after cooling , the mixture was filtered and the solution evaporated under reduced pressure . the residue was suspended in 1m hcl solution , filtered and washed with water and diethyl ether . the resulting solid was dried under vacuum to yield the title compound ( 4 . 0 g , 86 %). 1 h - nmr ( dmso - d6 ) δ 7 . 38 - 7 . 45 ( m , 3h ), 7 . 50 - 7 . 59 ( m , 3h ), 7 . 98 - 8 . 02 ( m , 2h ); ( m + 1 ) e / z 233 1 h - nmr ( dmso - d6 ) δ 7 . 24 ( m , 1h ), 7 . 42 ( m , 1h ), 7 . 62 - 7 . 60 ( m , 3h ), 8 . 04 ( d , 2h ); ( m + 1 ) e / z 235 1 h - nmr ( dmso - d6 ) δ 7 . 33 ( s , 1h ), 7 . 40 - 7 . 52 ( m , 6h ), 7 . 70 ( s , 1h ), 7 . 95 ( d , 2h ); ( m + 1 ) e / z 242 1 h - nmr ( dmso - d6 ) δ 2 . 29 ( s , 3h ), 7 . 31 - 7 . 50 ( m , 6h ), 7 . 83 ( dd , 1h ), 7 . 89 ( s , 1h ); ( m + 1 ) e / z 213 1 h - nmr ( dmso - d6 ) δ 7 . 47 - 7 . 55 ( m , 3h ), 8 . 1 ( d , 1h ), 8 . 11 - 8 . 16 ( m , 2h ), 8 . 32 ( dd , 1h ), 9 . 13 ( s , 1h ), 13 . 39 ( br s , 1h ); ( m + 1 ) e / z 200 a mixture of 4 - cyano - benzoic acid methyl ester ( 16 . 5 g , 102 mmol ), hydroxylamine hydrochloride ( 102 mmol ), nahco 3 ( 110 mmol ) in methanol ( 200 ml ) was stirred for 30 minutes at room temperature and heated to the reflux for a further 3 hours . after cooling , water ( 400 ml ) was added , the precipitate collected by filtration , washed and dried in a vacuum oven at 50 ° c . for 8 hours to give the title compound as a white solid ( 16 . 5 g , 83 %). ( m + 1 ) e / z 195 to a solution of 4 -( n - hydroxycarbamimidoyl )- benzoic acid methyl ester ( 5 . 7 g , 29 . 4 mmol ) in dioxane ( 30 ml ) was added cdi ( 1 . 2 eq ). the reaction mixture was heated to 110 ° c . for 30 minutes . after cooling the solvent was evaporated , the residue suspended in water and the ph adjusted to ph = 2 with aq . hcl ( 3m ). the precipitate was collected by filtration washed with water , suspended in aqueous solution of naoh ( 30 ml , 10 % w / w ) and methanol ( 50 ml ) and left stirring at room temperature overnight . after evaporation of the solvents , the residue was taken in water ( 30 ml ), ph adjusted to ph = 2 adding aq . hcl ( 3m ). the precipitate was collected by filtration , washed with water and dried under vacuum to yield the title compound as a white solid ( 4 . 1 g , 68 %). 1 h - nmr ( dmso - d6 ) δ 2 . 29 ( s , 3h ), 7 . 31 - 7 . 50 ( m , 6h ), 7 . 83 ( dd , 1h ), 7 . 89 ( s , 1h ); ( m + 1 ) e / z 213 to a solution of terephthalic acid monomethyl ester ( 5 g , 27 . 7 mmol ) in dichloromethane ( 40 ml ), cdi ( 27 . 7 mmol ) was added . after 10 minutes stirring , n - hydroxy - acetamidine ( 27 . 7 mmol ) was added and the resulting mixture stirred at room temperature for 3 hours . the solution was filtered and evaporated under reduced pressure to yield the title compound as a white solid ( 4 . 9 g , 75 %). a mixture of n -( 4 - methoxycarbonylbenzoyl ) oxy ) acetamidine ( 4 . 9 g , 20 . 7 mmol ) and sodium acetate ( 20 . 7 mmol ) in methanol ( 70 ml ) and water ( 20 ml ) was heated to 90 ° c . for 8 hours . after cooling a solid crystallised out of solution . the solid was filtered out , suspended in aq . naoh solution ( 10 % w / w , 30 ml ) and methanol ( 30 ml ) and left stirring at room temperature overnight . the solution was then evaporated under reduced pressure , the ph adjusted to ph = 3 adding aq . hcl ( 6m ). a precipitated formed , which was collected by filtration , washed with water , diethyl ether and dried under vacuum to yield the title compound as a white solid ( 2 . 5 g , 44 %). 1 h - nmr ( dmso - d6 ) δ 2 . 44 ( s , 3h ), 8 . 17 ( m , 4h ); ( m + 1 ) e / z 205 a mixture of 4 - cyano - benzoic acid methyl ester ( 4 . 02 g , 25 mmol ), sodium azide ( 32 . 5 mmol ) and triethylamine hydrochloride ( 32 . 5 mmol ) in toluene ( 40 ml ) is heated at 97 ° c . for 7 hours . after cooling the solution , water ( 100 ml ) was added . the aqueous phase was separated and to this solution hcl conc ( 7 g ) was added . a precipitate formed which was isolated by filtration and washed with water . the obtained solid was suspended in aq . naoh solution ( 20 ml , 10 % w / w ) and methanol ( 20 ml ) and left stirring at room temperature for 2 hours . the solvent was then evaporated , water was added to the residue and the ph acidified with hcl ( 6m ). a white precipitate formed which was isolated by filtration , washed with water and dried under vacuum to give the title compound ( 4 . 5 g , 95 %). to a solution of 4 -( n - hydroxycarbamimidoyl )- benzoic acid methyl ester ( 3 . 88 g , 20 mmol ) in dichloromethane ( 20 ml ), acetic anhydride ( 40 mmol ) was added . the mixture was left stirring at room temperature overnight . after 16 hours the solvent was evaporated , pyridine ( 30 ml ) was added and the reaction mixture heated at 95 ° c . for 2 days . after cooling the solution a solid crystallised out of solution . to this solution , water ( 20 ml ) was added and after 2 hours stirring at room temperature it was filtered and the solid collected . the solid was suspended in aq . naoh ( 30 ml , 10 % w / w ) and methanol ( 50 ml ) and left stirring at room temperature overnight . after evaporation of the solvents , the residue was taken in water ( 30 ml ), ph adjusted to ph = 2 adding aq . hcl ( 3m ). a precipitate formed which was collected by filtration , washed with water and dried under vacuum to yield the title compound as a white solid ( 3 . 8 g , 93 %). ( m + 1 ) e / z 205 . the biarylcarboxylic acids ( 0 . 00057 mol ) were treated with 5 ml of socl 2 for 5 h under reflux . the excess of socl 2 was removed by distillation and the crude acid chloride was used in the next reaction without further purification . a mixture of ( 4 - aryl - piperazin - 1 - yl )- alkylamine ( 0 . 3 mmol ), biarylcarboxylic acid chloride ( 0 . 3 mmol ), triethylamine ( 0 . 56 mmol ) and a catalytic amount of dmap in ch 2 cl 2 was stirred at 0 ° c . for 10 min then at room temperature for 4 h . the ch 2 cl 2 layer was washed with water , dried and concentrated . the residue purified by chromatography on silica gel with chcl 3 / meoh 95 / 5 as eluent to give the title compound . a solution of ( 4 - aryl - piperazin - 1 - yl )- alkylamine ( 0 . 00014 mol ) in 5 ml of dry ch 2 cl 2 was cooled to 0 ° c . the carboxylic acid ( 0 . 0002 mol ), 1 -( 3 - dimethylaminopropyl )- 3 - ethylcarbodiimide hydrochloride ( edc ) ( 0 . 0002 mol ) and a catalytic amount of dmap were added and the reaction mixture was stirred at room temperature for 16 h . the ch 2 cl 2 layer was then washed with water , dried and concentrated in vacuo and the residue purified by chromatography eluting with a gradient chcl3 / meoh 99 : 1 to 95 : 5 . to the preweighed acid ( 0 . 55 mmol ), dimethylformamide was added ( 2 ml ) to dissolve , followed by n , n ′- carbonyldiimidazole ( cdi ) ( 0 . 55 mmol ). the solution was then left for 60 minutes before adding the amine ( 0 . 6 mmol ) and the reaction was stirred for a further 16 hours . the solvent was removed under reduced pressure and the crude mixture was treated with 5 % meoh in dichloromethane ( 2 ml ) and washed with 10 % sodium hydroxide solution ( 2 ml ). this mixture was passed through a column packed with 5 grams of diatomaceous earth and the eluting the product with dichloromethane . the collected organic layer , containing the desired compound , was further purified using flash chromatography eluting with 10 % meoh in dichloromethane . fractions containing the product were combined and the solvent removed under reduced pressure . for less reactive carboxylic acids , activation was accomplished by heating the reaction at 60 ° c . for 2 h before adding the amine ( 1 eq ) ( 1m solution in dimethylformamide ) to the reaction mixture upon cooling ; the reaction is then shaken at room temperature for 18 - 24 h . alternatively , to a solution of carboxylic acid ( 0 . 3 mmol ) and cdi ( 0 . 3 mmol ) in acetonitrile ( 3 ml ), the amine ( 0 . 3 mmol ) was added after 10 minutes . the reaction mixture was exposed to microwave irradiation for 10 minutes at 100 ° c . after cooling the reaction mixture was absorbed on a scx cartridge , eluted with dichloromethane , methanol and methanol / ammonia solution . after evaporation , the residue was purified by silica column eluting with a gradient ethyl acetate / cyclohexane ( 1 : 1 ) ethyl acetate → ethyl acetate / methanol ( 9 : 1 ). the fractions containing the product were combined and the solvent evaporated . a solution of 4 - aminobutan - 1 - ol ( 20 . 71 g , 232 mmol ) in dichloromethane ( 50 ml ) was added to a stirring solution of 4 - bromobenzoyl chloride ( 51 g , 232 mmol ) in dichloromethane ( 250 ml ). diisopropylethylamine ( 40 . 4 ml , 232 mmol ) was added and the colourless solution was stirred at room temperature . lc / ms indicated completion of the reaction after 50 mins . the solution was transferred to a separating funnel and washed with water . a white solid precipitated out which was filtered off and washed with dichloromethane to afford pure product . the filtrate was treated with h 2 o which gave rise to further precipitate . the organic layer was washed with 1m hcl and nahco 3 ( sat ), dried over mgso 4 , filtered and concentrated in - vacuo to afford a further batch of product ( total yield 57 . 99 g ). a solution of oxalyl chloride ( 4 . 15 ml , 47 . 6 mmol ) in dichloromethane ( 200 ml ) was stirred under a n 2 flow at − 60 ° c . dmso ( 6 . 76 ml , 95 . 2 mmol ) was added cautiously ensuring that the temperature remained below − 50 ° c . after 15 mins a solution of 4 - bromo - n -( 4 - hydroxybutyl ) benzamide ( 10 g , 36 . 6 mmol ) in a mixture of dichloromethane ( 20 ml ), thf ( 40 ml ) and dmso ( 5 ml ) was added . after 30 mins the temperature had risen to − 50 ° c . after 1 h triethylamine ( 1 . 637 g , 16 . 18 mmol ) was added . the mixture was allowed to warm to room temperature and stirred overnight . lc / ms indicated completion of the reaction . h 2 o ( 200 ml ) was added to the reaction mixture . the organic layer was washed with 1m hcl , nahco 3 ( sat ) and brine , dried over mgso 4 , filtered and concentrated in - vacuo to afford an orange oil ( 9 . 93 g ). a solution of 4 - aminobutan - 1 - ol ( 20 . 3 g , 228 mmol ) in dichloromethane ( 50 ml ) was added to a stirring solution of 3 - bromobenzoyl chloride ( 50 g , 228 mmol ) in dichloromethane ( 250 ml ). dipea ( 39 . 6 ml , 228 mmol ) was added and the colourless solution was stirred at room temperature . lc / ms indicated completion of the reaction after 50 mins . the solution was transferred to a separating funnel and washed with water . a white solid precipitated out which was filtered off and washed with dichloromethane to afford pure product . the filtrate was treated with h 2 o which gave rise to further precipitate . the organic layer was washed with 1m hcl and nahco 3 ( sat ), dried over mgso 4 , filtered and concentrated in - vacuo to afford a further batch of product ( total yield 46 . 82 g , 76 %, 97 % pure by lc / ms ). a solution of oxalyl chloride ( 20 . 85 ml , 239 mmol ) in dichloromethane ( 900 ml ) was stirred under a n2 flow at − 60 ° c . dmso ( 33 . 9 ml , 478 mmol ) was added cautiously ensuring that the temperature remained below − 50 ° c . after 15 mins a solution of 3 - bromo - n -( 4 - hydroxybutyl ) benzamide 1 ( 50 g , 184 mmol ) in a mixture of dichloromethane ( 100 ml ), thf ( 400 ml ) and dmso ( 50 ml ) was added . after 30 mins the temperature had risen to − 50 ° c . after 1 h triethylamine ( 96 . 7 g , 956 mmol ) was added . the mixture was allowed to warm to room temperature and stirred overnight . lc / ms indicated completion of the reaction . h 2 o ( 1 l ) was added to the reaction mixture . the organic layer was washed with 1m hcl , nahco 3 ( sat ) and brine , dried over mgso4 , filtered and concentrated in - vacuo to afford an orange oil ( 9 . 93 g , & gt ; 100 %, 97 % pure by lc / ms ). to the preweighed amine ( 1 equivalent ), the aldehyde was added dissolved in anhydrous dichloromethane ( 1 . 2 eq , dichloromethane ). the solution was left to mix for 90 minutes before addition of sodium triacetoxyborohydride ( 1 . 5 equivalents ). the reaction was left to mix for a further 16 hours . the crude reaction was then washed with saturated nahco 3 ( 2 ml solution / reaction ) and the organic layer extracted . the dichloromethane crude solution was passed through an scx column , eluting the desired product in 20 % ammonia in methanol . fractions containing the compound were combined and the product purified further using hplc prep . general procedure for suzuki coupling of n -( 4 - amino ) butyl - 3 - or 4 - bromobenzamides — exemplified in detail for n -( 4 -( 4 - acetylpiperazin - 1 - yl ) butyl )- 4 - bromobenzamide and 2 - ethylphenylboronic acid n -( 4 -( 4 - acetylpiperazin - 1 - yl ) butyl )- 4 - bromobenzamide ( 86 mg , 0 . 225 mmol ) was dissolved in dme : etoh 1 : 1 ( 20 ml ) and added to a microwave tube containing 2 - ethylphenylboronic acid ( 34 mg , 0 . 225 mmol ). 1m na 2 co 3 in h 2 o was added ( 300 μl , 0 . 3 mmol ) followed by pd ( pph 3 ) 4 ( 26 mg , 0 . 0225 mmol ). the tube was capped , shaken by hand and loaded into the microwave for 10 mins at 150 ° c . the reaction was filtered through celite and washed with meoh . the filtrate was concentrated in - vacuo and purified by reverse phase preparative hplc . the product was taken on directly to form the hcl salt : 200 μl 1 . 25 m hcl in meoh and 800 μl dichloromethane were added to the title compound and the solution was shaken and concentrated in - vacuo to afford the hydrochloride salt ( 38 . 7 mg ). in dichloromethane at 0 ° c .- room temperature . a solution of aromatic amine ( 1 eq ) and triethylamine ( 1 eq ) in dichloromethane ( 0 . 2 mmol / ml ) is cooled at 0 ° c . under nitrogen atmosphere . 5 - bromopentanoyl chloride ( 1 eq ) in dichloromethane ( 0 . 3 mmol / ml ) is slowly added and the mixture stirred at room temperature for 1 . 5 hr . the amine ( 5 eq ) and triethylamine ( 1 eq ) are added at once and the reaction is stirred at room temperature for 40 hrs . the organic solution is then washed with brine , dried and the solvent removed . the product are crystallised by hexane : diethylether 1 : 1 or purified by flash chromatography . modified room temperature conditions for array synthesis : to a solution of aniline ( 1 eq ) and triethylamine ( 1 eq ) in dichloromethane ( 2 ml ) at room temperature was slowly added 5 - bromo - pentanoyl chloride ( 1 eq ) and the mixture stirred for 1 . 5 hr . the solution was added to a previously prepared vial containing the amine ( 5 eq ) and triethylamine ( 1 eq ) and the reactions were shaken at room temperature for 40 hrs . the organic solution was washed with brine , dried and the solvent removed . the products were purified by flash chromatography or by preparative hplc . in dichloroethane / dimethylformamide at 55 ° c . : a substituted aromatic amine ( 1 eq ) and triethylamine ( 1 eq ) are weighed in a glass vial and 1 , 2 - dichloroethane is added to give a 1 . 2 m solution ; 5 - bromovaleryl chloride ( 0 . 95 eq ) is then added dropwise as a solution in dimethylformamide ( 1 . 2 m ) and the reaction is shaken at room temperature for 1 h 30 min . the amine ( 3 eq ) and triethylamine ( 1 eq ) are then added as a solution in dce ( amine concentration 1 . 8 m ) and the reaction mixture shaken at 55 ° c . for 4 h . after this period , the reaction mixture is cooled and partitioned between water and dichloromethane ; the organic layer is washed with sat . nacl and dried over na 2 so 4 . the crude amides obtained after solvent evaporation at reduced pressure are purified by preparative hplc . prepared according the general procedure in dichloromethane at room temperature to give 3 . 7 g ( 70 %) of the title compound . c 16 h 24 n 3 obr mass ( calculated ) [ 354 . 29 ]; found [ m + h +]= 354 / 356 ( br ), nmr ( 400 mhz , dmso ): 1 . 43 ( 2h , m ); 1 . 55 ( 2h , m ); 2 . 23 ( 3h , s ); 2 . 27 - 2 . 50 ( 12h , m ); 7 . 44 ( 2h , d , j = 9 hz ); 7 . 55 ( 2h , d , j = 9 hz ); 10 . 05 ( 1h , s ). to a degassed mixture of 5 - alkylamino - pentanoic acid bromoaryl - amide ( 0 . 1 g , 1 eq ) and a substituted benzeneboronic acid ( 1 . 1 eq ) in acetonitrile / sodium carbonate 0 . 4 m solution 1 / 1 ( 4 ml ) a catalytic amount of pd [( pph 3 )] 4 ( 5 mmol %) was added . the reaction mixture was heated at 90 ° c . for 20 minutes under microwave irradiation ( 150 watt ) and then again other 20 minutes . the organic layer was separated and purified by scx column . the solvent was removed under reduced pressure to afford the corresponding product . to a cooled 0 . 2 m solution of amine ( 1 eq ) in dichloromethane , 1 eq of bromophenylisocyanate was added . the mixture was left stirring at 0 ° c . and it was stopped when a white solid was formed ( 1 h ), after ca . 1 hour . the product was recovered by filtration as a white solid which was used without further purification . general suzuki cross - coupling procedure for the synthesis of ureas microwave irradiation to a degassed 0 . 067 m solution of bromide ( 1 eq , prepared following the procedure for ureas described above ) in acetonitrile / water ( 1 / 1 ), the appropriate boronic acid ( 1 eq ) and na2co3 ( 3 eq ) were added followed by pd [( pph 3 )] 4 ( 10 % mol ). the solution was irradiated under microwave conditions , using the following parameters : power = 200 watt ; ramp time = 1 min ; hold time = 20 min ; temp = 90 ° c . ; pressure = 200 psi . the acetonitrile layer was separated and the crude mixture was purified using a scx column washing with dichloromethane / meoh followed by meoh and then nh3 / meoh to elute the product . the fractions containing the desired product were combined and dried under reduced pressure . the urea was weighted ( 1 eq , prepared following the procedure for ureas described above ), placed in a 2 - neck flask and dissolved in a degassed solution of acetonitrile / water ( 4 / 1 , 0 . 04 m ). to this solution boronic acid ( 1 . 1 eq ), na 2 co 3 ( 3 eq ) and pd [( pph 3 )] 4 ( 10 % mmol ) were added . the mixture was heated at 80 ° c . and stirred for 20 hours . the solution was filtered on celite layer and purified using scx or preparative hplc . prepared with a modification of pascal , j . c . ; et al . eur . j . med . chem ., 1990 , 25 , 291 - 293 : a solution of 1 . 48 g ( 0 . 0097 mol ) of 2 , 4 - dimethoxyaniline , 1 . 89 g ( 0 . 0160 mol ) of bis - 2 - chloroethylamine hydrochloride and 2 . 00 g of k 2 co 3 in 25 ml of 1 - butanol was refluxed for 24 h then filtered hot . the solvent was removed under reduced pressure and the residue triturated with acetone . the resulting powder was filtered and dried to give 1 . 25 g of the title compound . 1 h - nmr ( dmso - d 6 ) δ ( ppm ): 9 . 21 ( br s , 1h ); 6 . 82 ( d , 1h ); 6 . 52 ( s , 1h ); 6 . 42 ( d , 1h ); 3 . 74 ( s , 3h ); 3 . 68 ( s , 3h ); 3 . 12 ( s , 4h ); 3 . 07 ( s , 4h ). prepared following the general procedure outlined in nishikawa , y . ; et al ; chem . pharm . bull ., 1989 , 37 ( 1 ), 100 - 105 . a mixture of n -( 4 - bromobutyl ) phthalimide ( 0 . 00135 mol ), 1 -( 2 ′, 4 ′- dimethoxyphenyl )- piperazine hydrochloride ( 0 . 00135 mol ), k 2 co 3 ( 0 . 00270 mol ), nal ( 0 . 00186 mol ) and methylethyl ketone ( 7 ml ) was refluxed for 20 h with stirring . after the mixture had cooled , the insoluble materials were removed by filtration and washed with chcl 3 the filtrate and the washings were concentrated to dryness in vacuo . the residue was purified by cromatography on silica gel with chcl 3 / meoh 95 / 5 as eluent . yield : 68 %. 1 h - nmr ( cdcl 3 ) δ ( ppm ): 7 . 73 ( m , 4h ); 6 . 82 ( d , 1h ); 6 . 40 ( m , 2h ); 3 . 79 ( s , 3h ), 3 . 73 ( s , 3h ), 3 . 65 ( m , 2h ); 2 . 98 ( m , 4h ); 2 . 61 ( m , 4h ); 2 . 41 ( t , 2h ); 1 . 66 ( m , 4h ). a solution of 2 -{ 4 -[ 4 -( 2 , 4 - dimethoxy - phenyl )- piperazin - 1 - yl ]- butyl }- isoindole - 1 , 3 - dione ( 0 . 000236 mol ) and hydrazine hydrate ( 0 . 000478 mol ) in ethanol ( 2 ml ) was refluxed for 2 h with stirring . after the solution had cooled , any insoluble materials were removed by filtration and washed with etoh . the filtrate and the washings were concentrated in vacuo to dryness . the residue was taken up with chcl 3 . the chcl 3 layer was washed with water , dried and concentrated to give the title amine . yield : 50 %. 1 h - nmr ( cdcl 3 ) δ ( ppm ): 6 . 85 ( d , 1h ); 6 . 41 ( m , 2h ); 3 . 81 ( s , 3h ); 3 . 75 ( s , 3h ); 3 . 01 ( m , 4h ); 2 . 63 ( m , 4h ); 2 . 40 ( t , 2h ); 1 . 35 ( m , 6h ). prepared by reaction with 4 -( pyridin - 2 - yl )- benzoic acid according to the general procedure ( acid chloride method ). 1 h - nmr ( cdcl 3 ) δ ( ppm ): 8 . 66 ( d , 1h ); 8 . 02 ( d , 2h ); 7 . 85 ( d , 2h ); 7 . 75 ( m , 2h ); 7 . 23 ( m , 1h ); 6 . 96 ( br s , 1h ); 6 . 76 ( d , 1h ); 6 . 42 ( d , 1h ); 6 . 36 ( dd , 1h ); 3 . 78 ( s , 3h ); 3 . 72 ( s , 3h ); 3 . 47 ( m , 2h ); 2 . 97 ( m , 4h ); 2 . 65 ( m , 4h ); 2 . 47 ( t , 2h ); 1 . 70 ( m , 4h ) hplc : column zorbax c8 meoh 80 %/ h 2 o 20 %, 1 . 0 ml / min ; rt 6 . 54 ; area = 99 % prepared from 4 -[ 4 -( 2 , 4 - dimethoxy - phenyl )- piperazin - 1 - yl ]- butylamine and 4 - biphenylcarboxylic acid following the general procedure ( acid chloride method ). 1 h - nmr ( cdcl 3 ) δ ( ppm ): 7 . 82 ( d , 2h ); 7 . 5 - 7 . 6 ( m , 4h ); 7 . 48 - 7 . 5 ( m , 3h ); 6 . 89 ( br s , 1h ); 6 . 77 ( d , 1h ); 6 . 45 ( d , 1h ); 6 . 34 ( dd , 1h ); 3 . 80 ( s , 3h ); 3 . 73 ( s , 3h ); 3 . 49 ( m , 2h ); 2 . 96 ( m , 4h ); 2 . 64 ( m , 4h ); 2 . 45 ( t , 2h ); 1 . 68 ( m , 4h ). hplc : column : zorbax cn accn 40 %/ h 2 o ( cf 3 cooh ph = 2 . 3 ) 60 %, 0 . 8 ml / min ; rt = 5 . 396 ; area 98 % prepared from 4 -[ 4 -( 2 , 4 - dimethoxy - phenyl )- piperazin - 1 - yl ]- butylamine and 2 ′- nitrobiphenyl - 4 - carboxylic acid following the general procedure ( acid chloride method ). 1 h - nmr ( cdcl 3 ) δ ( ppm ): 7 . 7 - 7 . 9 ( m , 3h ); 7 . 45 - 7 . 55 ( m , 2h ); 7 . 3 - 7 . 4 ( m , 3h ); 6 . 84 ( br s , 1h ); 6 . 80 ( d , 1h ); 6 . 44 ( d , 1h ); 6 . 37 ( dd , 1h ); 3 . 80 ( s , 3h ); 3 . 74 ( s , 3h ); 3 . 49 ( m , 2h ); 2 . 97 ( m , 4h ); 2 . 63 ( m , 4h ); 2 . 46 ( t , 2h ); 1 . 68 ( m , 4h ) hplc : column zorbax cn meoh 50 %/ h 2 o ( cf 3 cooh ph = 2 ) 50 %, 0 . 4 ml / min ; rt = 17 . 209 ; area 88 % prepared from 4 -[ 4 -( 2 , 4 - dimethoxy - phenyl )- piperazin - 1 - yl ]- butylamine and 2 ′- fluorobiphenyl - 4 - carboxylic acid following the general procedure ( acid chloride method ). 1 h - nmr ( cdcl 3 ) δ ( ppm ): 7 . 81 ( d , 2h ); 7 . 56 ( d , 2h ); 7 . 1 - 7 . 4 ( m , 4h ); 6 . 99 ( s br , 1h ); 6 . 76 ( d , 1h ); 6 . 43 ( d , 1h ); 6 . 33 ( dd , 1h ); 3 . 78 ( s , 3h ); 3 . 71 ( s , 3h ); 3 . 46 ( m , 2h ); 2 . 94 ( m , 4h ); 2 . 60 ( m , 4h ); 2 . 44 ( t , 2h ); 1 . 66 ( m , 4h ) hplc : column zorbax cn accn 50 %/ h 2 o ( cf 3 cooh ph = 2 , 3 ) 50 %, 0 . 4 ml / min ; rt = 13 . 525 ; area 96 % prepared from 4 -[ 4 -( 2 , 4 - dimethoxy - phenyl )- piperazin - 1 - yl ]- butylamine and 2 ′- methylbiphenyl - 4 - carboxylic acid following the general procedure ( acid chloride method ). 1 h - nmr ( cdcl 3 ) δ ( ppm ): 7 . 80 ( d , 2h ); 7 . 35 ( d , 2h ); 7 . 2 - 7 . 4 ( m , 4h ); 6 . 88 ( br s , 1h ); 6 . 79 ( d , 1h ); 6 . 46 ( d , 1h ); 6 . 36 ( m , 1h ); 3 . 82 ( s , 3h ); 3 . 76 ( s , 3h ); 3 . 50 ( m , 2h ); 2 . 98 ( m , 4h ); 2 . 66 ( m , 4h ); 2 . 47 ( m , 2h ); 2 . 25 ( s , 3h ); 1 . 70 ( m , 4h ) hplc : column zorbax c8 accn 40 %/ h 2 o ( cf 3 cooh ph = 2 . 3 ) 60 %, 1 . 0 ml / min ; rt = 11 . 748 ; area 96 % 1h - nmr ( cdcl 3 ) δ ( ppm ): 7 . 72 ( m , 4h ); 6 . 89 ( m , 4h ); 3 . 81 ( s , 3h ); 3 . 69 ( t , 2h ); 3 . 15 ( m , 4h ); 2 . 60 ( 4h , m ); 2 . 40 ( t , 2h ); 1 . 66 ( m , 4h ). 1 h - nmr ( cd 3 od ) δ ( ppm ): 6 . 90 ( m , 4h ); 3 . 83 ( s , 3h ); 3 . 05 ( m , 4h ); 2 . 79 ( t , 2h ); 2 . 66 ( 4h , m ); 2 . 43 ( m , 2h ); 1 . 60 ( m , 4h ). prepared by reaction with 4 -( pyridin - 2 - yl )- benzoic acid according to the general procedure — carbodiimide method . 1 h - nmr ( cdcl 3 ) δ ( ppm ): 8 . 66 ( d , 1h ); 8 . 00 ( d , 2h ); 7 . 84 ( d , 2h ); 7 . 70 ( m , 2h ); 7 . 21 ( m , 1h ); 6 . 8 - 7 . 0 ( m , 5h ); 3 . 80 ( s , 3h ); 3 . 44 ( m , 2h ); 3 . 03 ( m , 4h ); 2 . 62 ( m , 4h ); 2 . 43 ( m , 2h ); 1 . 65 ( m , 4h ). hplc : column zorbax c8 meoh 80 %/ h 2 o 20 %, 0 . 8 ml / min ; rt = 4 . 72 ; area : 99 . 9 %. following the general procedure , 6 - indolecarboxylic acid ( 44 mg , 0 . 27 mmol ) is dissolved in dimethylformamide ( 1 ml ) and 1 , 1 ′- carbonyldiimidazole ( 44 mg , 0 . 27 mmol ) is added . 4 -[ 4 -( 2 , 4 - difluoro - phenyl )- piperazin - 1 - yl ]- butylamine ( 73 mg , 0 . 27 mmol ) dissolved in dimethylformamide ( 0 . 25 ml ) is then added and the mixture is allowed to react for 18 h . work - up followed by preparative hplc affords the title compound ( 51 mg , 41 %, & gt ; 95 % pure ) as formate salt . c 23 h 26 f 2 n 4 o mass ( calculated ) [ 412 . 49 ]; ( found ) [ m + h +]= 413 nmr ( 400 mhz , cdcl3 ): 1 . 51 ( 4h , m ); 2 . 34 ( 2h , t ); 2 . 47 ( 4h , bs ); 2 . 93 ( 4h , bs ); 3 . 26 ( 2h , m ); 6 . 49 ( 1h , s ); 6 . 95 - 7 . 01 ( 2h , m ); 7 . 12 - 7 . 17 ( 1h , m ); 7 . 40 ( 2h , m ); 7 . 6 ( 1h , dd , j = 8 . 4 , 1 . 2 ), 8 . 09 ( 1h , s ); 8 . 17 ( 1h , hcooh , s ); 8 . 26 ( 1h , t ); 11 . 27 ( 1h , s ). cdi ( 4 . 07 g , 25 mmol ) was added to a solution of 4 - pyridin - 2 - yl - benzoic acid ( 5 . 0 g , 25 mmol ) in dichloromethane and the reaction mixture stirred for 4 hours . 4 - aminobutanol ( 3 . 0 ml , 30 mmol ) was added and the reaction mixture stirred for 4 hours after which the solution was washed with a saturated solution of na 2 co 3 . the organic layer was separated , dried over mgso 4 , filtered and the solvent removed under reduced pressure . the product was purified by column chromatography ( dichloromethane , dichloromethane / meoh 1 %) to give 2 . 4 g of the title alcool . 1h nmr ( 400 mhz , dmso ): 8 . 71 - 8 . 66 ( 1h , m ), 8 . 53 - 8 . 46 ( 1h , m ), 8 . 78 ( 2h , d , 8 . 1 hz ), 8 . 12 ( 1h , d , 8 . 3 hz ), 7 . 94 ( 2h , d , 8 . 1 hz ), 7 . 92 - 7 . 83 ( 1h , m ), 7 . 46 - 7 . 36 ( 1h , m ), 4 . 38 ( 1h , t , 6 . 6 hz ), 3 . 42 ( 2h , dd , 6 . 6 hz , 12 . 0 hz ), 3 . 35 - 3 . 25 ( 2h , m ), 1 . 60 - 1 . 42 ( 4h , m ). a solution of oxalyl chloride ( 42 μl , 0 . 48 mmol ) in dichloromethane ( 5 ml ) was stirred under n 2 at − 60 ° c . dmso ( 34 μl , 0 . 48 mmol ) was added followed after 15 mins by a solution of alcohol ( 100 mg , 0 . 37 mmol ) in dichloromethane ( 100 ml ). after 2 h triethylamine ( 106 μl , 0 . 74 mmol ) was added . the mixture was then allowed to warm to room temperature and stirred overnight . lc / ms indicated completion of the reaction . the organic layer was washed with a saturated solution of nh 4 cl , dried over mgso 4 , filtered and concentrated under reduced pressure to give 100 mg of a white powder ( 92 % pure by lc / ms rt = 0 . 98 , m + 1 = 269 ) which was used in the next step without further purification . azepane ( 50 μl , 0 . 45 mmol ) was weighed into a clean glass vial . to this , the crude n -( 4 - oxo - butyl )- 4 - pyridin - 2 - yl - benzamide ( 100 mg , 0 . 37 mmol ) was added , dissolved in 2 ml of anhydrous dichloromethane . the reaction was left to mix for 90 minutes before addition of sodium triacetoxyborohydride ( 118 mg , 0 . 56 mmol ), after which it was stirred for 16 hours at room temperature before washing the crude reaction with saturated nahco 3 ( 2 ml solution ) and extracting the organic layer . the dichloromethane crude solution was passed through an scx column , eluting the desired product in 20 % ammonia in methanol . fractions containing the compound were combined and the product purified further using hplc prep to yield n -( 4 - azepan - 1 - yl - butyl )- 4 - pyridin - 2 - yl - benzamide as the formate salt ( 47 mg , 36 % yield ). 1 h nmr ( cdcl 3 ) 8 . 08 ( m , 4h ), 7 . 77 ( m , 3h ), 7 . 27 ( m , 1h ), 3 . 54 ( m , 2h ), 3 . 10 ( m , 6h ), 1 . 89 ( m , 6h ), 1 . 73 ( m , 6h ) following the general procedure in dichloroethane / dimethylformamide at 55 ° c ., 3 - chloroaniline ( 76 mg , 0 . 6 mmol ) and triethylamine ( 60 mg , 0 . 6 mmol ) are dissolved in dimethylformamide ( 0 . 5 ml ) and 5 - bromovaleryl chloride ( 113 mg , 0 . 57 mmol ) in dimethylformamide ( 0 . 5 ml ) is added dropwise . after 1 h 30 min , piperidine ( 153 mg , 1 . 8 mmol ) and triethylamine ( 60 mg , 0 . 6 mmol ) in dimethylformamide ( 0 . 5 ml ) and the reaction mixture heated at + 55 ° c . for 4 h . wok - up followed by preparative hplc affords the title compound ( 118 mg , 67 %) as a white solid as formate salt . c 16 h 23 c 1 n 2 o mass ( calculated ) [ 294 . 82 ]; ( found ) [ m + h +]= 295 nmr ( 400 mhz , dmso - d6 ): 1 . 48 ( 2h , m ); 1 . 52 ( 6h , m ); 2 . 31 ( 2h , t ); 2 . 48 ( 6h , m ); 7 . 05 ( 1h , dd , j = 8 , 1 . 2 ); 7 . 30 ( 1h , m ); 7 . 41 ( 1h , dd , j = 8 . 4 , 0 . 8 ); 7 . 80 ( 1h , s ); 8 . 21 ( 1h , hcooh , s ); 10 . 1 ( 1h , bs ). prepared according the general procedure in dichloromethane at room temperature to give 6 . 4 g ( 93 %) of the title compound . c 15 h 21 n 2 o 2 br mass ( calculated ) [ 341 . 24 ]; found [ m + h +]= 341 / 343 ( br ) nmr ( 400 mhz , dmso ): 1 . 44 ( 2h , m ); 1 . 57 ( 2h , m ); 2 . 29 ( 8h , m ), 3 . 54 ( 4h , m ), 7 . 44 ( 2h , d , j = 7 hz ), 7 . 54 ( 2h , d , j = 7 hz ). prepared according the general procedure in dichloromethane at room temperature to give 1 . 7 g ( 33 %) of the title compound . c 16 h 23 n 2 obr mass ( calculated ) [ 339 . 28 ]; found [ m + h +]= 339 / 341 ( br ), nmr ( 400 mhz , dmso ): 1 . 51 - 1 . 64 ( 10h , m ); 2 . 34 ( 2h , m ); 2 . 23 ( 2h , m ); 2 . 76 ( 4h , m ); 2 . 97 ( 2h , m ); 7 . 12 - 7 . 264 ( 2h , m ); 7 . 48 ( 2h , br d , j = 8 hz ); 7 . 97 ( 1h , s ). prepared according the general procedure in dichloromethane at room temperature followed by suzuki coupling to give 0 . 1 g ( 92 %) of the title compound . c 22 h 25 n 2 o 2 f 3 mass ( calculated ) [ 406 . 44 ]; ( found ) [ m + h +]= 407 nmr ( 400 mhz , dmso ): 1 . 45 ( 2h , m ); 1 . 6 ( 2h , m ); 2 . 3 ( 8h , m ); 3 . 55 ( 4h , m ); 7 . 21 ( 2h , d , j = 8 . 4 hz ); 7 . 36 ( 1h , d , j = 7 . 3 hz ); 7 . 56 ( 1h , m ); 7 . 63 ( 2h , d , j = 8 . 4 hz ); 7 . 68 ( 1h , m ); 7 . 79 ( 1h , d , j = 7 . 7 hz ) prepared according the general procedure in dichloromethane at room temperature followed by suzuki coupling to give 0 . 07 g ( 63 %) of the title compound . c 23 h 30 n 4 o 2 mass ( calculated ) [ 394 . 51 ]; ( found ) [ m + h +]= 395 nmr ( 400 mhz , dmso ): 1 . 43 ( 2h , m ); 1 . 58 ( 2h , m ); 2 . 10 ( 3h , s ); 2 . 12 - 2 . 44 ( 12h , m ); 7 . 40 ( 1h , s ); 7 . 49 ( 1h , m ); 7 . 68 ( 4h , m ); 7 . 78 ( 2h , m ); 8 . 06 ( 1h , s ); 8 . 11 ( 1h , s ); 9 . 97 ( 1h , s ). prepared according the general procedure in dichloromethane at room temperature followed by suzuki coupling to give 46 mg ( 51 %) of the title compound . c 24 h 31 n 3 o 3 mass ( calculated ) [ 409 . 53 ]; ( found ) [ m + h +]= 410 nmr ( 400 mhz , cd3od ): 1 . 62 ( 2h , m ); 1 . 74 ( 2h , m ); 2 . 07 ( 3h , s ); 2 . 41 - 2 . 49 ( 8h , m ); 3 . 53 ( 2h , m ); 3 . 58 ( 2h , m ); 3 . 78 ( 3h , s ); 6 . 98 ( 1h , m ); 7 . 04 ( 1h , d , j = 8 ); 7 . 27 ( 2h , m ); 7 . 43 ( 2h , d , j = 8 . 8 ); 7 . 56 ( 2h , d , j = 8 . 8 ) prepared according the general procedure in dichloromethane at room temperature followed by suzuki coupling to give 0 . 04 g ( 37 %) of the title compound . c 24 h 29 n 3 o 2 f 2 hco 2 h mass ( calculated ) [ 429 . 51 / 46 . 01 ]; ( found ) [ m + h +]= 430 . 28 nmr ( 400 mhz , dmso ): 1 . 44 ( 2h , m ); 1 . 58 ( 2h , m ); 1 . 66 ( 1h , m ); 1 . 75 ( 1h , m ); 1 . 96 ( 3h , s ), 2 . 32 ( 2h , m ); 2 . 42 ( 2h , m ); 2 . 52 ( 3h , m ); 2 . 62 ( 1h , m ); 3 . 54 ( 4h , m ), 7 . 24 - 7 . 42 ( 3h , m ); 7 . 5 ( 2h , d , j = 9 hz ); 7 . 7 ( 2h , d , j = 9 hz ); 8 . 16 ( 1h , s ); 10 . 03 ( 1h , s ) prepared according the general procedure in dichloromethane at room temperature followed by suzuki coupling to give 0 . 06 g ( 58 %) of the title compound . c 22 h 28 n 2 o 2 mass ( calculated ) [ 352 . 47 ]; ( found ) [ m + h +]= 353 . 32 nmr ( 400 mhz , dmso ): 1 . 34 ( 2h , m ); 1 . 40 - 1 . 47 ( 6h , m ); 1 . 57 ( 2h , m ); 2 . 19 - 2 . 33 ( 8h , m ); 6 . 73 ( 1h , d , j = 8 hz ); 6 . 95 ( 1h , s ); 6 . 99 ( 1h , d , j = 7 hz ); 7 . 23 ( 2h , m ); 7 . 32 ( 1h , m ); 7 . 51 ( 1h , d , j = 9 hz ); 7 . 87 ( 1h , s ); 9 . 56 ( 1h , br s ); 9 . 94 ( 1h , s ). 1 -( 4 - bromo - phenyl )- 3 -( 4 - morpholin - 4 - yl - butyl )- urea was weighed ( 0 . 8 g , 0 . 22 mmol ), placed in 2 necks flask and dissolved in a degassed solution of acetonitrile ( 4 ml ) and water ( 1 ml ). 2 - chloro - phenylboronic acid ( 0 . 33 g , 0 . 24 mmol ) and na2co3 ( 0 . 65 g , 0 . 6 mmol ) and a catalytic amount of pd [( pph 3 )] 4 werer then added in sequence and the mixture was heated at 80 ° c . and stirred for 20 hours . the solution was filtered on celite layer and purified using preparative hplc . c 21 h 26 c 1 n 3 o 2 mass ( calculated ) [ 387 . 91 ]; ( found ) [ m + h +]= 388 nmr ( 400 mhz , meoh ): 1 . 56 - 1 . 58 ( 2h , m ), 1 . 71 ( 2h , m ), 2 . 94 - 2 . 98 ( 2h , m ), 3 . 06 - 3 . 22 ( 4h , m ), 3 . 22 - 3 . 25 ( 2h , m ), 3 . 8 ( 4h , m ), 7 . 24 - 7 . 29 ( 5h , m ), 7 . 37 - 7 . 42 ( 3h , m ), 8 . 31 ( 1h , s ) table 1 shows a selection of the compounds synthesised , which were prepared according to the method indicated in the last column of the table and discussed in detail in the experimental procedures with the synthesis of examples 1 - 17 . when the compound is indicated as the hcl salt , the salt was formed by dissolution of the free base in methanol and addition of 1 eq 1m hcl in ether followed by evaporation of the solvents . when the compound is indicated as hcooh ( formic acid ) salt , the compound was purified by preparative hplc . cloning of alpha7 nicotinic acetylcholine receptor and generation of stable recombinant alpha7 nachr expressing cell lines full length cdnas encoding the alpha7 nicotinic acetylcholine receptor were cloned from a rat brain cdna library using standard molecular biology techniques . rat gh4c1 cells were then transfected with the rat receptor , cloned and analyzed for functional alpha7 nicotinic receptor expression employing a flipr assay to measure changes in intracellular calcium concentrations . cell clones showing the highest calcium - mediated fluorescence signals upon agonist ( nicotine ) application were further subcloned and subsequently stained with texas red - labelled a - bungarotoxin ( bgtx ) to analyse the level and homogeneity of alpha7 nicotinic acetylcholine receptor expression using confocal microscopy . three cell lines were then expanded and one characterised pharmacologically ( see table 2 below ) prior to its subsequent use for compound screening . a robust functional flipr assay ( z ′= 0 . 68 ) employing the stable recombinant gh4c1 cell line was developed to screen the alpha7 nicotinic acetylcholine receptor . the flipr system allows the measurements of real time ca 2 + - concentration changes in living cells using a ca 2 + sensitive fluorescence dye ( such as fluo4 ). this instrument enables the screening for agonists and antagonists for alpha 7 nachr channels stably expressed in gh4c1cells . gh4c1 cells stably transfected with rat - alpha7 - nachr ( see above ) were used . these cells are poorly adherent and therefore pretreatment of flasks and plates with poly - d - lysine was carried out . cells are grown in 150 cm 2 t - flasks , filled with 30 ml of medium at 37 ° c . and 5 % co 2 . ec 50 and ic 50 values were calculated using the idbs xlfit4 . 1 software package employing a sigmoidal concentration - response ( variable slope ) equation : the functional flipr assay was validated with the alpha7 nachr agonists nicotine , cytisine , dmpp , epibatidine , choline and acetylcholine . concentration - response curves were obtained in the concentration range from 0 . 001 to 30 microm . the resulting ec 50 values are listed in table 2 and the obtained rank order of agonists is in agreement with published data ( quik et al ., 1997 ). the assay was further validated with the specific alpha7 nachr antagonist mla ( methyllycaconitine ), which was used in the concentration range between 1 microm to 0 . 01 nm , together with a competing nicotine concentration of 10 microm . the ic 50 value was calculated as 1 . 31 ± 0 . 43 nm in nine independent experiments . functional flipr assays were developed in order to test the selectivity of compounds against the alpha1 ( muscular ) and alpha3 ( ganglionic ) nach receptors and the structurally related 5 - ht3 receptor . for determination of activity at alpha1 receptors natively expressed in the rhabdomyosarcoma derived te 671 cell line an assay employing membrane potential sensitive dyes was used , whereas alpha3 selectivity was determined by a calcium - monitoring assays using the native sh - sy5y cell line . in order to test selectivity against the 5 - ht3 receptor , a recombinant cell line was constructed expressing the human 5 - ht3a receptor in hek 293 cells and a calcium - monitoring flipr assay employed . the compounds were tested using the functional flipr primary screening assay employing the stable recombinant gh4c1 cell line expressing the alpha7 nachr . hits identified were validated further by generation of concentration - response curves . the potency of compounds from examples 1 - 254 as measured in the functional flipr screening assay was found to range between 10 nm and 30 microm , with the majority showing a potency ranging between 10 nm and 10 microm . the best exemplified compounds were also demonstrated to be selective against the alpha1 nach , alpha3 nach and 5ht3 receptors . neuroprotective activity of selected compounds was analyzed in an established cell - based assay of excitotoxicity induced by nmda in mixed primary rat cortical neurons as described previously ( stevens et al , 2003 ). in brief , test compounds were added 24 h before nmda application . incubation with nmda lasted 10 min or 24 h and cell mortality was assessed 24 h after application of the excitotoxic stimulus ( see fig1 ). selected compounds ( at concentrations ranging from 0 . 1 to 10 microm ) reduced mortality on average by 50 % and in some experiments a maximum of 80 % neuroprotection was observed . neuroprotective activity of compounds was analyzed in an in vivo animal model of cholinergic degeneration induced by quisqualic acid injection in the nucleus basalis of rats . subchronic treatment i . p . daily , for 7 days , with the compound at a dose of 3 mg / kg resulted in 60 % reduction in the degeneration of cholinergic neurons as demonstrated by determination of the number of chat - positive neurons ( a representative result is shown in fig2 ). cognitive behaviour was studied for selected compounds from example using the passive avoidance ( pa ) and object recognition ( ort ) tests in order to test the capability to reverse scopolamine - 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