AROMATIC SULFONAMIDE DERIVATIVES, THEIR USE AS ENZYME INHIBITORS AND PHARMACEUTICAL COMPOSITIONS CONTAINING THEM

Abstract: Aromatic sulfonamide derivatives, particularly benzene-sulfonamide, 4-fluorobenzenesulfonamide, 5-chloro-1-naphthalenesulfonamide and 5-isoquinolinesulfonamide derivatives are provided that inhibit Ca<2+>-dependent enzymes and proteins such as Phospholipase A2, protein kinases such as Protein Kinase C, and inhibit membrane fusion, thereby being a valuable drug for the treatment of inflammation, arthritis, infarction, nephritis and many other types of tissue injury.

Description: Aromatic sulfonamide derivatives, their use as enzyme inhibitors and pharmaceutical compositions containing them. This invention relates to aromatic sulfonamide derivatives, especially to benzenesulfonamide-, 4-fluorobenzenesulfon- amide-, 5-chloro-1-naphthalenesulfonamide- and 5-isoguino- linesulfonamide derivatives. This invention particularly relates to aromatic sulfonamide derivatives that inhibit Ca -dependent enzymes and proteins such as Phospholipase A2 / protein kinases such as Protein Kinase C, and membrane fusion, thereby being a valuable drug for the treatment of inflammation, arthritis, infarction, nephritis and many other types of tissue injury, and provides a process for the preparation thereof. The invention further relates to pharmaceutical compositions containing these derivatives. The invention is defined in detail in the claims. A groups in formula I-IV particularly include L-phenylalanine, -alanine, L-proline, L-valine, L-tryptophane and L-tyrosine - where the N atom is bound to the SO group. -• When the R j group in formula I-IV is hydrogen, then the R group is benzyl, biphenyl or a C3 Cg al ylene chain. A 6 embered heterocyclic ring (piperazine) may be formed through a ethylene group and adjacent nitrogen atoms. Exemplary benzenesulfonamide derivatives of the invention include: (1) N-beι__~me-"ulfαnyl-L-phenyM_nine piperazineamide [referred to as compound 1]; (2) N-beπzeπesulf onyl-L-alanine piperazineamide ' [referred, to as compound 2] ; (3) N-benzenesulfonyl-L-valine piperazineamide [referred to as compound.3 ; (4) N-beπzenesulfonyl-L-prOline piperazineamide [referred, to as compound 4]; (5) N-beι_εenesulfonyl-L-phenykl___ine-l,6-____ιι_nohex_neamide [referred to as compound 5]; (6) N-beι_-sne__ιlfonyl-L-phenykIa_ιine-l,3-ά_aminopropaneamide [referred to as compound 6]; (7) N-be___2nes-_fonyl-L-phenyk_anine biphenylamide [referred to as compound 7]; (8) N-benzenesulfonyl-L-alanine biphenylamide [referred to as compound 8 ; (9) N-benzenesulfonyl-L-valine biphenylamide [referred to as compound _]; (10) N-beπzenesulfony -L-proline biphenylamide [referred to as compound 101: (11) N-beπzenesuIfonyl-L-tryptόphane biphenylamide [referred to as compound 111: (12) N-beπzenesulfonyl-L-tyrosine biphenylamide [referred to as compound 121: Exemplary 4-fluorobenzenesu_fonylan_ide derivatives of the invention include: (13) N-(4- fluorotei-_£nesi-lfony_)-L-phenyIaIa___ne piperazineamide [referred to as compound 131: (14) N-(4-fluorobeπzeπesulf onyl) -L-alaπine piperazineamide [referred to as compound 141: (15) N-(4-fluorobeπzenesulfoπyI)-L-valine piperazineamide [referred to as compound 151: (16) N-(4-fluorobeπzenesuIfonyI)-L-proline piperazineamide [referred to as compound 161: (17) N-(4-fluorobenzenesuIfonyi)-L-phenyIalaniπe biphenylamide [referred to as compound 171: (18) N-(4-fIuorobenzenes_lfonyl)-L-aIanine biphenylamide [referred to as compound 18]; (19) N-(4-fluorobenzenesulfonyl)-L-valine biphenylamide [referred to as compound 19]; (20) N-(4-fIuorobenzenesulfonyl)-L-p-θ_ine biphenylamide [referred to as compound 201: Exemplary 5-cMoro-l-naphthalenesulfonamide derivatives of the invention include: (21) N-(5-chloro-l-naρhthalenesulfonyI)-L-phenylalaπine piperazineamide [referred to as compound 211: (22) N-(5-cMoro-l-ι pht_alene_αιlfonyι)-L-v_J___e piperazineamide [referred to as compound 22]; (23) N-(5-chloro-l-naphthaienesulfonyl)-L-proline piperazineamide [referred to as compound 23]; (24) N-{5-cWorcr-l-πiipht_aIenesulfonyi)-L-a-a-_ine piperazineamide [referred to as compound 24]' (25) N-(5-cMoro-l-π_ιpht_iilenesιu^onyl)-L-phenyIalan_ne-l,6-dia___nohexaneamide [referred to as compound 251: (26) N-(5-cMorc l-π_ιphthaIenesιrifonyl)-L-phenylalani_e biphenylamide [referred to as compound 26]; (27) N-(5-ctøoro-l-πapht_alenes_dfonyl)-L-alaπine biphenylamide [referred to as compound 27]; (28) -(5-chloro-l-naphthalenesulfonyI)-L-valine biphenylamide [referred to as compound 2!jS]; (29) N-(5-chloro-l-πaphthaleπesulfoπyl)-L-proIine biphenylamide [referred to as compound 29]; (30) N-biphenyiό-chloro-l-naphthaleπesulfoπamide [referred to as compound 301: Exemplary S-isoquinolinesulfonylamide derivatives of the invention include: (31) N-(5-__ q_-_ oline_n_lfonyl)-L-phenyl__lan_ne piperazineamide [referred to as compound 311: (32) N-(5-i_oqu_nolinesulfonyl)-L-valine piperazineamide [referred to as compound 321: (33) N-(5-isc_ιι_mo-_nesu_fonyl)-L-proline piperazineamide [referred to as compound 333; (34) N-(5-_soqu_nol_nesulfonyl)-L-a_anine piperazineamide [referred to as compound 341: __ (35) N-(5-__Oqu_nolinesutfonyl)-L-phenyl_laι_me-l 6-diaπ_ ohexaneaπιide [referred to as compound 351: (36) N-(5-__oq^ol_nesu-fonyl)-L-phenylalan_ne biphenylamide [referred to as compound 361: (37) N-(5-_sc_q^__-iolinesulfonyl)-L-v_line biphenylamide [referred to as compound 37]; (38) N-(5-isoquinol_nesulfonyl)-L-proline biphenylamide [referred to as compound 381: (39) N-(5-_-oqu_no_ine-_ myl)-L-_danine biphenylamide [referred to as compound 391: (40) N-(5-_3θquinoIinesulfonyl)-D-proline biphenylamide [referred to as compound 401: (41) N-biphenyl-5-iaxjuino_inesulfonan_ide [referred to as compound 411: (42) N-benzyl-5-isoq___ιolinesulfonamide [referred to as compound 421: The acid addition salts of the derivatives I-_L 13-16. 21-25. and __l-__5 according to this inventi are pharmaceutically acceptable non-toxic salts and can be prepared by conventional method Suitable examples of such pharmaceutically acceptable acid addition salts include the salts inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid and sulfuric acid; a the salts of organic acids such as acetic acid, citric acid, tartaric acid, lactic acid, succinic aci fumaric acid, maleic acid, methanesulfonic acid and p-toluenesulfonic acid. The benzenesulfonamide - and the 4-fluorobeι_zenesu_fonamide derivatives of formula I-II can prepared in accordance with the following equations: wherein A is a part of an amino acid, R j is a hydrogen atom, R2 is a biphenyl group and R3 is hydrogen or a fluor atom; or VIII wherein A is a part of an amino acid, R^ is a hydrogen atom and R2 a C3 or Cg alkyleπe chain; or R j and 2 are linked directly to form a piperazine ring. R3 is a hydrogen or a fluor atom and X a protective group. Derivative V can be prepared as follows: - IX wherein AA is an amino acid and R3 is a hydrogen or a fluor atom. The πaphthalenesulfonylamide derivatives of formula III can be prepared in accordance to the following equations: wherein A is a part of an amino acid or a single bond, R^ is a hydrogen atom and R2 is a biphenyl group; or XII XIII wherein A is a part of an amino acid, R^ is a hydrogen atom and R2 a Cg alkylene chain; or j and R2 are linked directly to form a piperazine ring. X is a protective group. Derivative XI can be prepared as follows: XAA Jl' HN XAN AN XIV VI XV XI wherein AA is an amino acid, R j is a hydrogen atom, R2 a biphenyl group and X a protective group. Derivative XII can be prepared as follows: *\ XAN — AN > R,X XVI XII wherein AA is an amino acid, R^ is a hydrogen atom and R2 a Cg alkylene chain; or R^ and 2 are linked directly to form a piperazine ring. X are protective groups. The .5C<j__nolinesuIfonamide derivatives of formula IV can be prepared in accordance to t following equations: wherein A is a part of an amino acid or a single bond, R^ is a hydrogen atom and R2 a phenyl- o biphenyl group; or CB-Z= carboxybenzyl BOC= t-butoxycaroonyl wherein A is a part of an amino acid, R^ is a hydrogen atom and 2 a alkylene chain; or ^ and R2 are linked directly to form a piperazine ring. X is a protective group. Exemplary compounds of formula V include N-beπzenesulfonyl-L-phenylalanine, N- benzenesuIfonyl-L-alanine, N-benzenesulfonyl-L-valine, N-beπzenesulfonyl-L-proline, N- benzenesulfonyl-L-tryptophane and N-beπzenesulfonyl-L-tyrosine; N-(4-_luorobenzenesulfonyl)-L- phenylalanine, N-(4-fluorobenzenesuIfoπyl)-L-aIanine, N-(4-fluorobeπzenesulfonyl)-L-valine, N- (4-fIuorobeπzenesulf onyl) -L-proline. Exemplary compounds of formula VT include 4-aminobiphenyI and 4-aminobenzyl. Exemplary compounds of formula VII include N-CBZ-piperazine, N-BOC-l,6-diaminohex_ne and N-CBZ-l,3-diaminopropane. Exemplary compounds of formula XI include L-phenylalaniπe biphenylamide, L-alanine biphenylamide, L-valin biphenylamide, L-prolin biphenylamide and D-pro line biphenylamide. Exemplary compounds of formula XII include L-phenylalaπine-N-CBZ-piperazineamide, L- alaπine-N-CBZ-piperazineamide, L-valine-N-CBZ-pϊperazineamide and L-phenylalanine-N-BOC- 1,6-diaminohexaneamide. The reaction between the compound of formula V and the compound of formula VI, respectively VII is best carried out in presence of dicyclohexylcarbodiimide (DCC) and a reaction medium like dimethylformamide (DMF) or dioxane; 1-hydroxy- benzotriazole is used to avoid racemization . The amount of the compound of formula V preferably is the same as of the compound of formula VI or VII. The amount of DCC is preferably about 1 to 5 equivalents and more preferably 1 to about 2 equivalents for each mol of the compound of formula V. The amount of 1-hydroxybenzotriazole is preferably about 1 to 5 equivalents and more preferably 1 to about 2 equivalents for each mol of the compound of formula V. The reaction between the compound of formula V and VI respectively VII can be carried out typically at a temperature of about -10"C to about 60"C and preferably from about O'C to 30 * C The reaction time which can be employed is typically about lh to about 24h and preferably from lh to about 5h. The method of obtaining the compounds of formula I and II from VIII may vary depending upon the protective group of X selected, generally known methods can be employed in this invention. For example when the protective group of X is an alkyloxycarbonyl group such as t- butoxycarbonyl, the desired products can be obtained by hydrolysis with an acid. When the protective group of X is an arylmethyloxycarbonyl group such as benzyloxycarbonyl, the desired compounds can be obtained by hydrogenation or hydrolysis with an acid. The reaction between the compound of formula X and XI respectively XII is best carried out in presence of an acid acceptor. Exemplary acid acceptors which can be employed include alkali metal compounds such as hydroxide, bicarbonate or carbonate, e.g. sodium hydroxide, potassium hydroxide, sodium bicarbonate, sodium carbonate, potassiumcarbonate and tertiary amines such as triethylamine and pyridine. In general the reaction is carried out in presence of a reaction medium. Exemplary reaction media which can be employed include ethers, such as dioxane or THF and halogenated hydrocarbons such as CHC1 3 and CH 2 C1 2 . The amount of the compound of formula X preferably is the same as of the compounds of formula XI or XII. The amount of the acid acceptor employed is preferably about 2 to 5 equivalents and more preferably about 2 to 3 equivalents for each mol of the compound of formula X. The reaction between the compound of formula X and XI respectively XII can be carried out typically at a temperature of about lO'C to about 60 * C and preferably at 20 * C to 30'C. The reaction time which can be employed is typically about lh to about 24h and preferably from lh to about 5h. The method of obtaining the compound of formula III from XIII can be carried out under the same conditions as the reaction from the compound of formula VIII to the compound of formula I and II. The reaction between the compound of formula XVII and XI respectively XII can be carried out under the same condition as the reaction between the compound of formula X and XI respectively XII except that the amount of the compound of formula XVII preferably is 0.9 equivalents of th compound of formula XI respectively XII and that the reaction temperature is preferably of abou O'Cto SO'C. It has now been found that the derivatives of formula I-IV (and their pharmaceutically acceptabl salts if applyable) have pharmacologically and biochemically interesting properties such as Phospholipase 2 (PL 2) inhibition activity. The effect of the derivatives of formula I-IV of this invention on PLA2 can be proved in vitro by taking bovine pancreatic PL 2, l-stearoyl-2-[C^J- arachidonyl-l-phosphatidylcholine and CaCl2 and adding a derivative of the formula I-IV resulting in inhibition of the PL 2. When for example N-(5-cUoro-l-__ιpht_alenesι__ nyl)-L-pheny_ϋ_ιιine- 1,6-diaminohexaneamide, i.e. compound 25 was added and a complete inhibition was designed 100%, the concentration which would bring about an inhibition of 50%, Le. IC50 was 67μM. The following examples illustrate the present invention in more detail, but they are given for illustrative purposes only and are not to be construed as limiting the invention. Synthesis of the Precursors Example 1 Equimolar amounts of O-beπzyl-L-tyrosine and benzeπesulfonyl chloride were mixed in IN NaOH (200 mol %) and stirred for 2h. After 0.5h the desired compound precipitated. The solution was adjusted to pH 2 with 2N HCl and filtered- The precipitate was washed several times with H2O, dried in vacuo over P2O5 and crystallized to yield the desired compound PL SubstantiaEy the same procedure as described above was repeated except that L-tryptophane was used in place of the L-tyrosine derivative to give P2. The yields of the reactions and the analytical data of the two compounds are given in table 1. Example 2 Equimolar amounts of L-phenyialanine and p-fluorobeπzenesulfonyl chloride were mixed in IN NaOH (200 mol %) and stirred at 60'C for 2h. The solution was adjusted to pH 2 with 2N HCl and extracted with EtOAc. The organic phase was dried over MgSOψ filtered, evaporated and the residue crystallized to yield P3. Substantially the same procedure as described above was repeated except that L-alanine (→P4). L- Yaline (— »P5) and L-proline (— . P6) were used in place of L-phenylalanine. The yields of the reactions and the analytical data of the different compounds are given in table 1. Example 3 To equimolar amounts of N-t-BOC-L-phenylalanine and 4-aminobiphenyl in DMF (ca 0.5M) 1 hydroxybenzotriazole (150 mol %) was added. The mixture was cooled to 0 * C and DCC (110 mol %) was added in one portion. After stirring for lh at 0"C and lh at 25 * C the reaction mixture was filtered and the DMF evaporated in vacuo. The ' residue was taken up in EtOAc (ca 0.1M) and washed subsequently with saturated NaHQ. 3 solution, 2N citric acid and H2O. After drying over MgSOψ filtering and evaporation the residue was crystallizecL The compound obtained above was dissolved in CH2CI2 (ca 0.4M) cooled with ice and the same amount of TFA added. After lh the solvents were evaporated and the residue crystallized: P7. Substantially the same procedure as described above was repeated except that N-t-BOC-L-valine (→P8) was used in place of the L-phenylalanine derivative. When N-CBZ-L-proline was used as starting material the deprotection was done by overnight hydrogenation with H2, 10% Pd/C in EtOH (- * P9). In case of N-t-BOC-L-alanine the free amine was obtained after evaporation of the solvents and stirring the residue with saturated NaHCU3 solution (→PIO). The yields of the reactions and the analytical data of the different compounds are given in table 2. Example 4 Basically the same procedure as in example 3 except that N-CBZ-piperazine [1] and N-BOC-6- aminohexane were used in place of 4-aminobiphenyl. In case of N-CBZ-piperazine N-t-BOC protected amino acids and in case of N-t-BOC-6-aminohexane N-CBZ protected amino acids were used. Deprotection of the N-t-BOC group was done as described before with TFAJ H- ^ at 0 * C (→P11,P12,P13,P14) and of the N-CBZ group with H 2 . 10% Pd/C in EtOH (→P15,P16). The free amine was dissolved in acetone, equimolar amounts of oxalic acid in acetone added, the precipitate filtered and crystallized. The yields of the reactions and the analytical data of the different compounds are given in table 2. Synthesis of the Compounds Example 5 To equimolar amounts of N-benzenesulfoπyl-L-phenylalanine [2] and N-CBZ-piperazine [1] in DMF (ca 0.5M) l-hydrox * benzotriazole (150 mol %) was added. Th. mixture was cooled to 0"C and DCC (110 mol %) was added in one portion. After stirring for lh at 0"C and lh at 25"C the reaction mixture was filtered and the DMF evaporated in vacuo. The residue was taken up in EtOAc (ca O.LM) and washed subsequently with saturated NaHCO3 solution, 2N citric acid and H2O. After drying over MgSOψ filtering and evaporation of the solvent the residue w crystallized (P17). The compound obtained above was dissolved in abs. EtOH (ca 0.1M), 10% Pd/C (10 weight added and stirred overnight in a H atmosphere. The reaction mixture was filtered over celite an the pH of the solution adjusted to 2 with cone. HCl. Evaporation 'of the solvent yielded a whit residue which was crystallized: 1. Substantially the same procedure as described above was repeated except that N-beπzenesulfonyl L-alanine [3](→2) . N-benzenesulfonyl-L-valine [4](→P18 →j_) and N-benzenesulfonyl-L-prolin [5](- * > P19 —»4) were used in place of N-benzenesulfonyl-L-phenylalaπine. The yields of the reactions and the analytical data of the different compounds are given in table (intermediates) and table 4 and 8 (compounds). Exam i? 6 Basically the same procedure as in example 5 except that N-t-BOC-6-aminohexane (—> P20 —>S) o N-CBZ-amiπopropane [6](-→ P21 →6) were used in place of N-CBZ-piperazine. Deprotection o the N-t-BOC group was done with TFA/CH 2 Cl2 at 0 * C and of the N-CBZ group with H 2 , 10 Pd C in EtOH as described before. The yields of the reactions and the analytical data of the two compounds are given in table 3 (intermediates) and table 4 and 9 (compounds). Example 7 To equimolar amounts of N-benzenesulf onyl-L-phenylalanine [2] and 4-aminobiphenyl in DMF (ca 0.5M) 1-hydroxybenzotriazole (150 mol %) was added. The mixture was cooled to 0 * C and DCC (110 mol %) was added in one portion. After stirring for lh at 0"C and lh at 25"C the reaction mixture was filtered and the DMF evaporated in vacuo. The residue was taken up in EtOAc (ca 0.1M) and washed subsequently with saturated NaHCC^ solution, 2N citric acid and H2O. After drying over MgSOφ filtering and evaporation of the solvent the residue was crystallized: 7. Substantially the same procedure as described above was repeated except that N-beπzenesulfonyl- L-alanine [3](-→S), N-benzenesulfonyl-L-valine [4] (→9_), N-benzenesulfonyl-L-proline [5] (-. 12) and N-beπzenesulf onyl-L-tryptophane P2 (-» 11) were used in place of N-benzenesulfonyl-L- phenylalanine. The yields of the reactions and the analytical data of the different compounds are given in table 4,10,11 and 12. Example 8 Basically the same procedure as in example 7 except that O-_«n_yl-N-benzenesulfonyl-L-tyrosin PI was used in place of N-benzenesulfonyl-L-phenylalanine (-. P22). The compound obtained above was dissolved in p-dioxane ^O HOAc 15:1:1 (ca lOmM), 10 Pd C (10 weight %) added and the mixture stirred overnight in a H2 atmosphere. The reactio mixture was filtered over celite and the solvents evaporated in vacuo. The residue was crystallized The yields of the reactions and the analytical data of the two compounds are given in table (intermediate) and table 4 and 10 (compound). Example 9 I To equimolar amounts of N-p-fluorobenzenesulfonyl-L-phenylalanine P3 and N-CBZ-piperazine i DMF (ca 0.5M) l-hydroxybenzotriazole (150 mol %) was added. The mixture was cooled to 0 * and DCC (110 mol %) was added in one portion. After stirring for lh at 0 * C and lh at 25 * C th reaction mixture was filtered and the DMF evaporated in vacuo. The residue was taken up i EtOAc (0.1M) and washed subsequently with saturated NaHCO3 solution, 2N citric acid and H2O. After drying over MgSO 4 , filtering and evaporation of the solvent the residue was crystallized: P23. The compound obtained above was dissolved in abs. EtOH (ca O.IM) 10% Pd/C (10 weight %) added and the mixture stirred overnight in a H2 atmosphere. The reaction irώcture was filtered over celite and the pH of the solution adjusted to 2 with cone. HCl. Evaporation of the solvent yielded a white residue which was crystallized: 13. Substantially the same procedure as described above was repeated except that N-p- fluorobenzenesulfonyl-L-alanine P4 (→ P24 -» 14). N-p-fluorobeπzenesulfoπyl-L-valine P5 (-. P25 -» 15) and N-p-fluorobenzenesulfonyl-L-proline P6 (— . P26 — » 16) were used in place of P3. The yields of the reactions and the analytical data of the different compounds are given in table 3 (intermediates) and 5 and 13 (compounds). Example 10 To equimolar amounts of N-p-fluorobenzenesuifonyl-L-phenylalanine P3 and 4-aminobiphenyl in DMF (ca 0.5M) l-hydroxybenzotriazole (150 mol %) was added. The mixture was cooled to 0 * C and DCC (110 mol %) was added in one portion. After stirring for lh at 0 * C and lh at 25 * C the reaction mixture was filtered and the DMF evaporated in vacuo. The residue was taken up in EtOAc (ca 0.1M) and washed subsequently with saturated NaHC03 solution, 2N citric acid and H2O. After drying over MgSO 4 , filtering and evaporation of the solvent the residue was crystallized: 17. Substantially the same procedure as described above was repeated except that N- fluorobenzenesulfonyl-L-alanine P4 (-. 18). N-p-fluorobenzenesulfonyl L-valine P5 (—>1~) and p-fluorobenzenesulfonyl L-proline P6 (-. 20) were used in place of P3. The yields of the reactions and the analytical data of the different compounds are given in table and 14. Example 11 To a concentrated solution of the amine Pll in p-dioxane and IN NaOH (200 mol %) an equimola amount of 5-chloro-l-naphthalenesu_fonyI chloride [7,8] was added and the suspension stirred fo 3h_ Acidification with cone. HCl to pH 2, extraction with CH2CI2 (3x), washing of the organi phase once with brine, drying over MgSOψ filtering and evaporation of the solvents in vacu yielded a residue which was purified by flash chromatography on silica gel (solvent system hexane/EtOAc 1:1). The compound obtained above was dissolved in abs. EtOH (ca 50mM) 10% Pd C (10 weight %) added and the mixture stirred for 3 days in a H2 atmosphere. The reaction mixture was filtered over celite, the solvent evaporated in vacuo, the residue purified by flash chromatography on silica gel (solvent system C^C^/MeOH 5:1) and crystallized: 21. Substantially the same procedure as described above was repeated except that the amine P12 (→ 22) 2 , P13 (→23J 3 or P14 (-. 24) 4 were used in place of the amine Pll. The yields of the reactions and the analytical data of the different compounds are given in table 6 and 15. Example 12 To a concentrated solution of the amine P15 in p-dioxane and IN NaOH (200 mol %) an equimolar amount of 5-chIoro-l-πaphthaienesulfonyl chloride was added and the suspension stirred for 3h. H2O was added and the mixture extracted thrice with CH2CI2. The organic phase was washed once with H2O, dried over MgSOψ filtered and the solvents evaporated in vacuo. Flash chromatography on silica gel (solvent system hexane EtOAc 1:2) yielded the pure intermediate which was dissolved in CH2CI2 (ca 50mM) and the same volume of TFA added. After lh the solvents were evaporated in vacuo, the residue dissolved in a small volume of H2O and the pH of the solution adjusted to 8 with solid NaH ^. Extraction with fourportioπs of Q^C^/ eOH 3:1, washing of the organic phase with H2O, drying over MgSOψ filtering and evaporation of the solvents in vacuo yielded a compound which was purified by flash chromatography on silica gel (solvent system C^C^/MeOH/concN^ 20:2:1) and crystallized as HCl-salt 2~. The yield of the reaction and the analytical data of the compound are given in table 6 and 16. 2 Solvent system for me intermediate: e___e/_-OAc 1: 1; no chromatography- was necessary for 22- ^ Solvent system for the intermediate: hexane EtOAc 1:2: solvent system for 2: C^Ch/McOH 10:3. Solvent system for the intermediate hexan cfi&OAc 1 :2; no chrcmatogr_p_y was necessary for 24 Example 13 To a concentrated solution of the amine P7 in THF and IN NaOH (200 mol %) an equimola amount of 5-chloro-l-πaphthalenesulfonyl chloride was added and the suspension stirred for 3 H2O was added and the mixture extracted thrice with CH2CI2. The organic phase was washed onc with H2O, dried over MgSO.4, filtered and the solvents evaporated in vacuo. Flash chromatograph on silica gel (solvent system hexane EtOAc 4:1) and crystallization yielded the pure compound 26. Substantially the same procedure as described above was repeated except that the amine P1 (-»27P was used in place of the amine P7. The yields of the reactions and the analytical data of the two compounds are given in table 6 and 17 Example 14 To a concentrated solution of the amine P8 in p-dioxane was added IN NaOH (200 mol %). Th precipitate partly dissolved after addition of a concentrated solution of an equimolar amount of 5 chloro-1-naphthalenesulfonyl chloride in THF. After 15min_ the desired compound precipitated After 2h H2O was added and the suspension filtered, the solid dried and crystallized twice to obtai the pure compound 28. Substantially the same procedure as described above was repeated except that the amine P9 (-»29) was used in place of the amine P8. The yields of the reactions and the analytical data of the two compounds are given in table 6 and 17. Example 15 To a concentrated solution of 4-aminobiphenyI in p-dioxane was added IN NaOH (200 mol %) and an equimolar amount of 5-chloro-l-naphthalenesulfonyl chloride. After stirring for 2h the solvents were evaporated in vacuo. The residue was purified by flash chromatography on silica gel (solvent system hexane EtOAc 2:1) and crystallized to yield 20- The yield of the reaction and the analytical data of the compound are given in table 6 and 17. Example 16 To an ice cooled suspension of 5-isoquinolinesulfonyl chloride hydrochloride [9] in CH2CI2 was added NEt 3 (220 mol %). To the yellow solution the amine Pll (90 mol %) in CΑ-£--2 was ------ dropwise. After lOmin. the cooling was removed and the stirring continued for 2 The pH of the solution was adjusted to 7-8 with a saturated NaHCU3 solution. The phases were separated and the organic phase washed once with H2O, dried over MgSOφ filtered and the solvent evaporated in 1 Solvent svstem for 22: hexane EtOAc 2:1. vacuo. Flash chromatography on silica gel (solvent system C^C^/MeOH 100:3) yielded a white foam. A solution of the compound obtained above in 25% HBr HOAc (8ml mmol) was stirred under ice cooling for 5h_ After addition of Ε -p (ca 40ml mmol) the precipitate was filtered and dissolved in a few ml of H2O. The solution was slightly basified with IN NaOH and extracted with C^C^/MeOH 3:1 (4x). The organic phase was washed once with H2O, dried over MgSO.4, filtered and the solvents evaporated in vacuo. Flash chromatography on silica gel (solvent system CH 2 Cl2 MeOH conc. NH3 20:4:1) yielded a foam which was dissolved in EtOH and treated with cone. HCl to give the salt 21- Substantially the same procedure as described above was repeated except that the amine P12 (→32) 5 ' 7 P13 (→ 13) and P14 (→3_4) 8 ' 9 were used in place of the amine Pll. The yields of the reactions and the analytical data of the different compounds are given in table 7 and 18. Example 17 To an ice cooled suspension of 5-isc uiπolinesulfbnyl chloride hydrochloride in CH2CI2 was added NE-3 (220 mol %). To the yellow solution the amine P15 (90 mol %) in CH 2 C1 2 was added dropwise. After lOmin. the cooling was removed and stirring continued for 2h. A saturated NaHC03 solution was added to adjust the pH to 7-8. The phases were separated and the organic phase washed once with H2O, dried over MgSOφ filtered and the solvent evaporated in vacuo. The residue was purified by flash chromatography on silica gel (solvent system C^Cl^-eOH 100:4). To the compound obtained above, dissolved in CH2C-2 (ca 50mM), the same volume of TFA was added. After lh the solvents were evaporated in vacuo, the residue dissolved in a very small volume of H2O and the pH of the solution adjusted to 8 with solid NaHCθ3. The H2O phase was extracted with CH2Cl2/MeOH 3:1 (4x). The organic phase was dried over MgSOψ filtered and evaporated to dryness. The residue was purified by flash chromatography on silica gel (solvent system CH 2 Cl2/MeOH/conc. NH3 20:4:1) and crystallized as HCl-salt 35, The yield of the reaction and the analytical data of the compound are given in table 7 and 16. Example 18 To an ice cooled suspension of 5-isoquinoIinesulfonyl chloride hydrochloride in CH2CI2 was added NEt 3 (220 mol %). To the yellow solution the amine P7 (90 mol %) was added dropwise. After lOmin. the cooling was removed and stirring continued for 2h. The pH of the solution was adjusted - Solvent system for the intermediate: C^Cl j TfeOH 100:4. 22. ύt EtOH was treated with cooc. HCL The precipitate was filtered and thoroughly washed with ElOH. The sait could noi be crystallize-L 8 Solvent system for the intermediate: CHICIT M-OH 100:5. * * was not purified by flash chromatography but directly crystallized as HQ-salt. to 7-8 with a saturated NaHCO.3 solution. The phases were separated and the organic phase washe once with H2O, dried over MgSO_ { , filtered and the solvent evaporated in vacuo. Flas chromatography on silica gel (solvent system CH2CΪ2/MeOH 100:3) yielded a white solid whic was crystallized to give 3_6_. Substantially the same procedure as described above was repeated except that the amine P8 (- * >37) P9 (→__-D 10 , P10 HIS!) 11 and (+)-P9 (→40) were used in place of the amine P7. The yields of the reactions and the analytical data of the different compounds are given in table and 19. Example 19 Substantially the same procedure as described in example 18 was used except that 4-aminobipheny ■ →41) I 2 and 4-aminobenzol (-. 42) were used in place of the amine P7. The yields of the reaction and the analytical data of the two compounds are given in table 7 and 19. Enzyme Assay Phospholipase A2 0? LA2) activity was measured using bovine pancreatic PLA2 as the enzyme an a sonicated dispersion of l-stearoyl-2-[C-'- 4 ]-arachidonyl-phosphatidylcholine (56mCi/mmol) as th substrate in the following manner [11,12]: 60ng bovine pancreatic PLA2 was mixed and incubate with buffer and inhibitors (lOOmM Tris, pH 8; lOOmM CaCl 2 ; 20mM EDTA, pH 8) at 37 * C fo lOmin. to allow interaction between the enzyme and the drugs. The substrate (38nCi) in buffer an 0.3% cholate was added to initiate the reaction which continued for 20miiL at 37'C. The total reaction volume was 0.1ml. The reaction was stopped by adding 0.1ml of an ice cold EtOH/HOA 98:2 mixture. Released arachidonic acid was separated from the unreacted substrate via thin laye chromatography on silica gel (solvent system CHCl3/MeOH/H2θ 14:6:1). The radioactivity of th two spots was quantified using a bioscanner. Inhibitors were dissolved in buffer ^ and tested in duplicate within each experiment, and eac inhibitor was tested in at least two experiments. When necessary, inhibitors were dissolved in DMSO "" . The % inhibition at a given concentration was combined for several experiments and the IC5 Q determined from a semilog plot of % inhibition vs concentration. Under the condition described the rate of hydrolysis was 20-25% of the substrate being hydrolyzed in absence of inhibitors. The results are listed in table 20. 10 Solvent system for 12 and _i: CHTCIVMCOH 100:2. 11 Solvent system for 39: CH 2 α 2 ΛieOH 100:5. 12 Solvent system for L and _2: C ^Cl-McOH 100:2. • - 0.01ml inhibitor solution were used in these experiments. 0.005ml inhibitor solution wen: used in these experiments (= 5 vol 9c). Literature [1] L.Goldman & J.H.Williams, J.Org-Chem. 18,815 (1953) [2] W.H.SchuUer & CNiemanπ, JACS 73,1644 (1951) [3] TJr.Buckley 8~ HLRapoport, JACS 101,6157 (1981) [4] CCKnudsen & H.Rapoport, J.Org.Chem.4&2260 (1983) [5] KRapoport et al. JACS 10_5_.1095 (1984) [6] GJAtwell & W.ADeπny, Synthesis 1032 (1984) [7] CSkopp 8~ G-Schwenker, Arc Pharm. 317,649 (1984) [8] RGHart et al. Methods in Enzymology 102,195 (1983) [9] H.Hidaka, Eur.Pa_eπt 0187371 (1985) [10] T.Matsui etaL .Soc.Org.Syn_h_Chem Japan 11,320 (1985) [11] RAIonso et al. Biochim.Biophys.Acta 878273 (1986) [12] W.CRipka et al. J.CelLBiochem.40,279 (1989) Table 8: 'H-NMIl (300 MHz. O .O) Table 9; l-NMR (300 Mllz, D 2 0) Tnble 10: 'H-NMR (300 MHz, acclonc-d, (7) or DMSO-d, (12)) Tiililc II: 'll-NMR (300 Mil/., acclono-il,) Table 12: 'H-NMR (300 MHz, DMSO-d 6 ) Table 14: 'H-NMR (300 MHz. acetonc-d 6 ) Table 14 conl,: Table 15: 'H-NMR (300 MHz. DMSO-d, (21). D.O) doubling of I he signals Table 15 conl.: Table 16: 'H-NMR (300 MHz, D.O) Table 17: "ll-NMR (300 MI . aceloncd^. CDCI, (43)) Table 17 conl.: O D O S. y _> -α -. oό co 00 Ov -- p- Table 18 conl. Table 19: 'll-N Il (300 Mil-, acelono-d,) Table 19 conl.: Hf Table 20 11 Compounds are only listed if their IC 50 is lower than 600μM. 21 Promotion of the hydrolysis rate at lOOmM: 138% of the value in absence of an inhibitor. 31 Promotion of the hydrolysis rate at lOOmM: 125% of the value in absence of an inhibitor. 4 Inhibition at lOOμM: 70.1%, at 300μM 54.0%. 51 Inhibition at lOμM: 31.6%, at lOOμM 28.3%.

Claims: CLAIMS 5 1. Aromatic sulfonamide derivatives of the formula wherein 5 Z is phenyl, naphthyl, (5)- or (δ)-isoquinolyl, possibly substituted by halogen A is a single bond or an amino acid group where the N atom of the amino acid group is bound to SO2 and its carboxyl group to the N π atom of the above identified general formula R-| is hydrogen, and R- is biphenyl, a C 2 to C g alkylene group; or is pheny.l if Z is neither naphthyl nor chloronaphthyl, and A is not a single bond; or 5 R. and R together form the piperazine ring, and its pharmaceutically acceptable non-toxic acid additional salts with inorganic or organic acids. o 2. Derivatives of claims 1 having one of the formulas wherein A, R, and R, have the meaning as defined in claim 1 , Y is F, Cl, Br, being the same or different, and n is 0, 1 or 2. 54 3. Derivatives of claim 1 or 2, wherein A is the L-phenylalanine, L-alanine, L-proline, L-valine, L-tryptophane or L-tyrosine group. 4. Derivates of claim 1 or 2, wherein A is the L-phenylalanine, L-alanine, L-valine or L-proline group. 5. Derivates of claim 1 or 2, wherein A is the L-phenylalanine group. 6. Derivates of any of the preceding claims, wherein R-| is hydrogen, R 2 is biphenyl. 7. Derivates of any of the claims 1 to 5, wherein R and R together form the piperazine ring. 1 2 8. Derivates of any of the claims 1 to 5, wherein R-| is hydrogen, and R 2 is C, or CL alkylene. 9. Derivates of any of the claims 1 to 5, wherein A is a single bond Ri is hydrogen, and R 2 is biphenyl. 1 10. Derivates of any of claims 2 to 5 with formula III, wherein c Y is F or Cl in 5-position n is 1 R- | is hydrogen- and R is biphenyl. 10 11. Derivates of any of claims 2 to 5 with formula IV, wherein Y is hydrogen n is 0 R-| is hydrogen, and - c R is biphenyl. 12. Use of the compounds of any of the preceding claims in¬ cluding the derivates of formula I, wherein A is a single bond, Z naphthyl or chloronaphthyl, . hydrogen, 0 and R phenyl, *** 2+ as inhibitors of Ca - dependent enzymes and proteins. 13. Use according to claim 12 as inhibitors of Phospholipase A p and protein kinases. 5 14. A pharmaceutical composition for use as claimed in any of the claims 12 and 13 with pharmaceutically accep¬ table non-toxic addition salts including the salts of inorganic acids, namely hydrochloric acid, phosphoric 0 acid and sulfuric acid; and the salts of organic acids namely citric acid, tartaric acid and methanesulfonic acid. 5

Inventors: DUMONT RAYMOND, DUMONT, RAYMOND
Applicants: PHARNO WEDROPHARM GMBH, PHARNO-WEDROPHARM GMBH
Application year: 1991
Main classification: C07C311
IPC codes: A61K31, A61P13, A61P19, A61P29, A61P37, A61P43, C07C311, C07D207, C07D209, C07D217, C07D295, C07D401, C07D403, C12N9