Patent Application: US-74298808-A

Abstract:
the invention relates to compounds , in particular 2 - amino - 3 , 4 , 5 ,- trisubstituted thiophenes , pharmaceutical compositions containing them and the uses of said compounds and compositions for diseases related to sphingosine - 1 - phosphate receptors , predominantly s1p3 receptors . the diseases include cardiovascular diseases , atherosclerosis , cancer , pulmonary oedema , autoimmune disorders and adult respiratory distress syndrome .

Description:
the compounds of the present invention ( formula 1 ) may be prepared by several synthetic procedures . for example , the synthetic route to obtain some 2 - amino - 3 , 4 , 5 ,- trisubstituted thiophene derivatives ( formula 2 ) is depicted in the scheme herein below : according to this scheme , the appropiate carbonyl compounds were reacted with benzoylacetonitrile derivatives and sulfur in ethanol in the presence of diethylamine to provide the products . these benzoylacetonitrile derivatives , when not commercially available , were synthesized by condensation of the acetonitrile anion with the appropiate substituted methyl benzoates . this invention is further described in the following specific examples , which do not limit the scope of the invention described in the claims . the examples detailed here of the general formula ( 2 ) are synthesised according to the route detailed in scheme 1 . chemicals and solvents all reagents were obtained from commercial sources and all solvents were of an analytical grade . chromatography thin - layer chromatography ( tlc ) was carried out using merck silica gel plastic backed f 254 plates , visualised under uv ( 254 nm ). instruments and analysis elemental analyses were performed for c , h , n ( leiden institute of chemistry , leiden university , the netherlands ). 1 h and 13 c nmr spectra were recorded on a bruker ac 200 ( 1 h nmr , 200 mhz ; 13 c nmr , 50 . 29 mhz ) spectrometer with tetramethylsilane ( tms ) as an internal standard . chemical shifts are reported in ppm ( δ ) relative to this . melting points were determined on a büchi melting point apparatus and are uncorrected . mass spectra were measured on a finnigan mat tsq - 70 spectrometer equipped with an electrospray interface for esi experiments . spectra were collected by constant infusion of the analyte dissolved in methanol . esi is a soft ionisation technique resulting in protonated , sodiated species in positive ionisation mode and deprotonated species in the negative ionisation mode . general procedure for the synthesis of 2 - amino - 3 - benzoylthiophenes . to a suspension of 5 mmol of carbonyl compound , 5 mmol of benzoylacetonitrile derivative , and 5 . 05 mmol of sulfur in 1 . 5 ml of ethanol was added 1 ml of diethylamine . the mixture was stirred for 2 h at 50 ° c . the mixture was cooled to room temperature . if the product crystallized from the crude reaction mixture , the precipitate was collected and recrystallized from the appropiate solvent . when no crystallization occurred , the mixture was evaporated , purified by column chromatography and crystallized from the appropiate solvent . ( 2 - amino - 4 , 5 - dimethyl - 3 - thienyl )( phenyl ) methanone was prepared as described above , starting from methyl ethyl ketone and benzoylacetonitrile : mp 133 - 134 ° c ., 1 h nmr ( cdcl 3 ) δ 1 . 54 ( s , 3h ch 3 ), 2 . 13 ( s , 3h , ch 3 ), 6 . 40 ( bs , 2h , nh 2 ), 7 . 39 - 7 . 54 ( m , 5h , h arom ). ( 2 - amino - 4 , 5 - dimethyl - 3 - thienyl )( 3 - chlorophenyl ) methanone was prepared as described above , starting from methyl ethyl ketone and 3 - chlorobenzoylacetonitrile : mp 114 - 116 ° c ., 1 h nmr ( cdcl 3 ) δ 1 . 52 ( s , 3h ch 3 ), 2 . 10 ( s , 3h , ch 3 ), 6 . 75 ( bs , 2h , nh 2 ), 7 . 30 - 7 . 49 ( m , 4h , h arom ). ( 2 - amino - 4 , 5 - dimethyl - 3 - thienyl )( 4 - chlorophenyl ) methanone was prepared as described above , starting from methyl ethyl ketone and 4 - chlorobenzoylacetonitrile : mp 123 - 125 ° c ., 1 h nmr ( cdcl 3 ) δ 1 . 55 ( s , 3h ch 3 ), 2 . 13 ( s , 3h , ch 3 ), 7 . 35 - 7 . 49 ( m , 6h , nh 2 , h arom ). ( 2 - amino - 4 - ethyl - 5 - methyl - 3 - thienyl )( phenyl ) methanone was prepared as described above , starting from 3 - pentanone and benzoylacetonitrile : mp 129 - 131 ° c ., 1 h nmr ( cdcl 3 ) δ 0 . 71 ( t , 3h ch 3 ), 2 . 09 ( q , 2h , ch 2 ), 2 . 16 ( s , 3h , ch 3 ), 6 . 10 ( bs , 2h , nh 2 ), 7 . 30 - 7 . 65 ( m , 5h , h arom ). ( 2 - amino - 4 - ethyl - 5 - methyl - 3 - thienyl )[ 3 -( trifluoromethyl ) phenyl ] methanone was prepared as described above , starting from 3 - pentanone and 3 - trifluoromethylbenzoylacetonitrile : mp 100 - 101 ° c ., 1 h nmr ( cdcl 3 ) δ 0 . 68 ( t , 3h ch 3 ), 2 . 00 ( q , 2h , ch 2 ), 2 . 16 ( s , 3h , ch 3 ), 6 . 55 ( bs , 2h , nh 2 ), 7 . 54 - 7 . 80 ( m , 4h , h arom ). ( 2 - amino - 4 - ethyl - 5 - methyl - 3 - thienyl )( 3 - chlorophenyl ) methanone was prepared as described above , starting from 3 - pentanone and 3 - chlorobenzoylacetonitrile : mp 99 - 102 ° c ., 1 h nmr ( cdcl 3 ) δ 0 . 71 ( s , 3h ch 3 ), 2 . 08 ( q , 2h , ch 2 ), 2 . 13 ( s , 3h , ch 3 ), 6 . 40 ( bs , 2h , nh 2 ), 7 . 30 - 7 . 60 ( m , 4h , h arom ). ( 2 - amino - 4 - ethyl - 5 - methyl - 3 - thienyl )( 4 - chlorophenyl ) methanone was prepared as described above , starting from 3 - pentanone and 4 - chlorobenzoylacetonitrile : mp 108 - 110 ° c ., 1 h nmr ( cdcl 3 ) δ 0 . 71 ( s , 3h ch 3 ), 2 . 09 ( q , 2h , ch 2 ), 2 . 16 ( s , 3h , ch 3 ), 6 . 00 ( bs , 2h , nh 2 ), 7 . 36 - 7 . 55 ( m , 4h , h arom ). ( 2 - amino - 4 , 5 , 6 , 7 - tetrahydrobenzo [ b ] thiophen - 3 - yl )( phenyl ) methanone was prepared as described above , starting from cyclohexanone and benzoylacetonitrile : 152 - 153 ° c ., 1 h nmr ( cdcl 3 ) δ 1 . 40 - 1 . 62 ( m , 2h ch 2 ), 1 . 68 - 1 . 80 ( m , 4h , 2 × ch 2 ), 2 . 48 - 2 . 55 ( m , 2h , ch 2 ), 6 . 65 ( bs , 2h , nh 2 ), 7 . 38 - 7 . 60 ( m , 5h , h arom ). ( 2 - amino - 4 , 5 , 6 , 7 - tetrahydrobenzo [ b ] thiophen - 3 - yl )( 2 - chlorophenyl ) methanone was prepared as described above , starting from cyclohexanone and 2 - chlorobenzoylacetonitrile : mp 145 - 147 ° c ., 1 h nmr ( cdcl 3 ) δ 1 . 44 - 1 . 71 ( m , 6h , 3 × ch 2 ), 2 . 44 - 2 . 49 ( m , 2h , ch 2 ), 7 . 21 - 7 . 37 ( m , 4h , h arom ). ( 2 - amino - 4 , 5 , 6 , 7 - tetrahydrobenzo [ b ] thiophen - 3 - yl )[ 3 -( trifluoromethyl ) phenyl ] methanone was prepared as described above , starting from cyclohexanone and 3 - trifluoromethylbenzoylacetonitrile : mp 122 - 123 ° c ., 1 h nmr ( cdcl 3 ) δ 1 . 41 - 1 . 63 ( m , 2h , ch 2 ), 1 . 68 - 1 . 79 ( m , 4h , 2 × ch 2 ), 2 . 47 - 2 . 54 ( m , 2h , ch 2 ), 7 . 00 ( bs , 2h , nh 2 ), 7 . 48 - 7 . 72 ( m , 4h , h arom ). ( 2 - amino - 4 , 5 , 6 , 7 - tetrahydrobenzo [ b ] thiophen - 3 - yl )( 3 - chlorophenyl ) methanone was prepared as described above , starting from cyclohexanone and 3 - chlorobenzoylacetonitrile : mp 114 - 115 ° c ., 1 h nmr ( cdcl 3 ) δ 1 . 42 - 1 . 58 ( m , 2h , ch 2 ), 1 . 70 - 1 . 87 ( m , 4h , 2 × ch 2 ), 2 . 47 - 2 . 55 ( m , 2h , ch 2 ), 6 . 88 ( bs , 2h , nh 2 ), 7 . 31 - 7 . 44 ( m , 4h , h arom ). ( 2 - amino - 4 , 5 , 6 , 7 - tetrahydrobenzo [ b ] thiophen - 3 - yl )( 3 - iodophenyl ) methanone was prepared as described above , starting from cyclohexanone and 3 - iodobenzoylacetonitrile : mp 160 - 162 ° c ., 1 h nmr ( cdcl 3 ) δ 1 . 47 - 1 . 82 ( m , 6h , 3 × ch 2 ), 2 . 49 - 2 . 55 ( m , 2h , ch 2 ), 6 . 80 ( bs , 2h , nh 2 ), 7 . 14 - 7 . 79 ( m , 4h , h arom ). ( 2 - amino - 4 , 5 , 6 , 7 - tetrahydrobenzo [ b ] thiophen - 3 - yl )[ 4 -( trifluoromethyl ) phenyl ] methanone was prepared as described above , starting from cyclohexanone and 4 - trifluoromethylbenzoylacetonitrile : mp 145 - 147 ° c ., 1 h nmr ( cdcl 3 ) δ 1 . 48 - 1 . 50 ( m , 2h , ch 2 ), 1 . 69 - 1 . 76 ( m , 4h , 2 × ch 2 ), 2 . 48 ( m , 2h , ch 2 ), 6 . 91 ( bs , 2h , nh 2 ), 7 . 54 - 7 . 69 ( m , 4h , h arom ). ( 2 - amino - 4 , 5 , 6 , 7 - tetrahydrobenzo [ b ] thiophen - 3 - yl )( 4 - chlorophenyl ) methanone was prepared as described above , starting from cyclohexanone and 4 - chlorobenzoylacetonitrile : mp 140 - 142 ° c ., 1 h nmr ( cdcl 3 ) δ 1 . 47 - 1 . 52 ( m , 2h , ch 2 ), 1 . 71 - 1 . 81 ( m , 4h , 2 × ch 2 ), 2 . 47 - 2 . 54 ( m , 2h , ch 2 ), 6 . 71 ( bs , 2h , nh 2 ), 7 . 34 - 7 . 45 ( m , 4h , h arom ). ( 2 - amino - 4 , 5 , 6 , 7 - tetrahydrobenzo [ b ] thiophen - 3 - yl )( 4 - bromophenyl ) methanone was prepared as described above , starting from cyclohexanone and 4 - bromobenzoylacetonitrile : mp 152 - 153 ° c ., 1 h nmr ( cdcl 3 ) δ 1 . 46 - 1 . 54 ( m , 2h , ch 2 ), 1 . 67 - 1 . 83 ( m , 4h , 2 × ch 2 ), 2 . 46 - 2 . 51 ( m , 2h , ch 2 ), 6 . 80 ( bs , 2h , nh 2 ), 7 . 33 - 7 . 55 ( m , 4h , h arom ). ( 2 - amino - 4 , 5 , 6 , 7 - tetrahydrobenzo [ b ] thiophen - 3 - yl )( 4 - iodophenyl ) methanone was prepared as described above , starting from cyclohexanone and 4 - iodobenzoylacetonitrile : mp 150 - 152 ° c ., 1 h nmr ( cdcl 3 ) δ 1 . 41 - 1 . 87 ( m , 6h , 3 × ch 2 ), 2 . 43 - 2 . 52 ( m , 2h , ch 2 ), 6 . 82 ( bs , 2h , nh 2 ), 7 . 17 - 7 . 75 ( m , 4h , h arom ). ( 2 - amino - 4 , 5 , 6 , 7 - tetrahydrobenzo [ b ] thiophen - 3 - yl )( 4 - methylphenyl ) methanone was prepared as described above , starting from cyclohexanone and 4 - methylbenzoylacetonitrile : mp 155 - 156 ° c ., 1 h nmr ( cdcl 3 ) δ 1 . 45 - 1 . 50 ( m , 2h , ch 2 ), 1 . 70 - 1 . 91 ( m , 4h , 2 × ch 2 ), 2 . 38 ( s , 3h , ch 3 ) 2 . 47 - 2 . 54 ( m , 2h , ch 2 ), 6 . 60 ( bs , 2h , nh 2 ), 7 . 16 - 7 . 42 ( m , 4h , h arom ). ( 2 - amino - 4 , 5 , 6 , 7 - tetrahydrobenzo [ b ] thiophen - 3 - yl )( 4 - nitrophenyl ) methanone was prepared as described above , starting from cyclohexanone and 4 - nitrobenzoylacetonitrile : mp 202 - 204 ° c ., 1 h nmr ( cdcl 3 ) δ 0 . 85 - 1 . 16 ( m , 6h , 3 × ch 2 ), 1 . 88 - 2 . 00 ( m , 2h , ch 2 ), 6 . 43 ( bs , 2h , nh 2 ), 7 . 01 - 7 . 69 ( m , 4h , h arom ). methyl 4 [( 2 - amino - 4 , 5 , 6 , 7 - tetrahydrobenzo [ b ] thiophen - 3 - yl )- carbonyl ] benzoate was prepared as described above , starting from cyclohexanone and methyl 4 -( 2 - cyanoacethyl ) benzoate : mp 159 - 160 ° c ., 1 h nmr ( cdcl 3 ) δ 1 . 54 ( s , 3h , ch 3 ), 2 . 13 ( s , 3h , ch 3 ), 6 . 40 ( bs , 2h , nh 2 ), 7 . 39 - 7 . 54 ( m , 5h , h arom ). 4 -[( 2 - amino - 4 , 5 , 6 , 7 - tetrahydrobenzo [ b ] thiophen - 3 - yl )- carbonyl ] benzoic acid was prepared as described above , starting from cyclohexanone and methyl 4 -( 2 - cyanoacetyl ) benzoate and hydrolisation : mp 232 - 233 ° c ., 1 h nmr ( cd 3 od ) δ 1 . 40 - 1 . 49 ( m , 2h ch 2 ), 1 . 64 - 1 . 70 ( m , 4h , 2 × ch 2 ), 2 . 42 - 2 . 48 ( m , 2h , ch 2 ), 7 . 43 - 8 . 09 ( m , 4h , h arom ). ( 2 - amino - 4 , 5 , 6 , 7 - tetrahydrobenzo [ b ] thiophen - 3 - yl )( 3 , 4 - dichlorophenyl ) methanone was prepared as described above , starting from cyclohexanone and 3 , 4 - dichlorobenzoylacetonitrile : mp 152 - 154 ° c ., 1 h nmr ( cdcl 3 ) δ 1 . 46 - 1 . 54 ( m , 2h , ch 2 ), 1 . 67 - 1 . 83 ( m , 4h , 2 × ch 2 ), 2 . 45 - 2 . 51 ( m , 2h , ch 2 ), 7 . 11 ( bs , 2h , nh 2 ), 7 . 27 - 7 . 56 ( m , 3h , h arom ). a primary function of certain cell surface receptors is to activate second messenger systems upon binding of an agonist . the s1p 1 receptor selectively binds to and activates g i - proteins and activation of this receptor subtype thus results in the inhibition of camp production . the s1p 2 receptor predominantly activates g q - proteins finally resulting in increases in intracellular calcium . s1p 3 receptor activation results in both , an inhibition of camp production and an increase in intracellular calcium , via the activation of g i - and g q - proteins accordingly , we performed measurements on camp accumulation ( lance camp 384 kit , perkin elmer , zaventem , belgium ) and measurements on changes in intracellular calcium ( for method see jongsma et al ., 2006 ) ( 8 ). in table 1 results of the camp accumulation assay at the s1p 1 and s1p 3 receptor are displayed for a number of compounds , in comparison with results for s1p itself . compounds with efficacy at the s1p 2 receptor also induced s1p 2 - mediated changes in intracellular calcium but generally with lower potency ( data not shown ). none of the compounds tested showed efficacy at the s1p 2 receptor . 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