NL194549C - Compounds with a smooth muscle relaxation activity of blood vessels and pharmaceutical compositions containing these compounds. - Google Patents

Description

1 194549

Compounds with a smooth muscle relaxation activity of blood vessels and pharmaceutical compositions containing these compounds

The invention relates to new isoquinoline sulfonamides with a relaxation activity for the smooth muscles of blood vessels and a low toxicity. The invention further relates to pharmaceutical compositions containing these compounds.

In particular, the invention relates to compounds with a relaxation activity for the smooth muscle cells of blood vessels.

Quinoline compounds with relaxation activity for blood vessel smooth muscle are described, for example, in Japanese Unexamined Patent Publications (Kokai) Nos. 60-81168, 61-126026, 61-271221, 61-293914, 62-103066 and 63-211267 and U.S. Patent Nos. 4,634,770, 4,678,783, 4,709,032 and 4,798,897.

The European patent application 0061673 relates to 5-isochinoin sulfonamides, which compounds are provided with a ring structure adjacent to the SO2 group, which compounds therefore have a different basic structure than the compounds described in the present invention, which compounds do not have a ring structure adjacent to the SO2 group.

Chemical Abstracts 103,160,407 m (1985), extract from JP Kokai 60/81168, relates to 5-isochinoin sulfonamides provided with a ring structure adjacent to the SO2 group, while the compounds of the present invention relate to quinoline sulfonamine derivative derivatives that do not have a ring structure adjacent to the SO 2 groups.

European patent application 0330910 relates to 5-iso-quinoin sulfonamides that contain a ring structure adjacent to the SO2 group, while quinoline sulfonamido derivatives having a ring structure adjacent to the SO2 group are excluded from the present invention.

Chemical Abstracts (CA) 212215u describes two compounds, namely W-7 and HA = -1004 25 H

HA-1004 SO 2 -N-CH 2 CH 2 -N-C-CH 2

II

Isoquinoline NH

30 H

W-7 SO 2 -N- (CH 2) 6-NH 2

Isoquinoline in which both the compound W-7 and HA-1004 do not have a branched structure, namely the structure in which the carbon atom adjacent to the nitrogen atom is provided with four different substituent groups. In addition, both compounds do not have a phenyl group, naphthyl group or heterocyclic group.

From European patent application 0109023 an isoquinoline sulfonamide derivative is known which can be represented schematically with the following formula: H / ^ Ar SO 2 -N-CH no.4 I \ ^

45 Isoquinoline CH 2 NH-C

\ nhr3

The meaning of R 4 and / or R 3 mentioned in this European patent application precludes the formation of the 50 N-containing ring structure according to the present isoquinoline sulfonamide derivative.

Of the compounds described in the above references, some have adequate relaxation activity with regard to smooth muscles, but present problems with regard to toxicity, specific action with regard to certain organs and safety.

The objectives of the present invention are to obtain new compounds with a sufficient relaxation activity for the smooth muscles and a low toxicity, a high specific action with regard to certain organs and with a high degree of safety.

The invention provides isochilin sulfonamides as described in the preamble, which are characterized in that these compounds correspond to formula 1, on the formula sheet wherein Y represents N or H 3 C-N 1, and R 1 represents a hydrogen atom, an optionally substituted lower alkyl group, a formyl group , a halophenyl-propargyl group, an optionally substituted aralkyl group or optionally substituted phenyl groups, and R2 represents a group represented by formula 3 on the formula sheet, wherein R10 represents a hydrogen atom, a nitro group, an optionally substituted amino group, an optionally substituted hydroxyl group, a lower alkyl group or represents a halogen atom, or R 11 and R 10 together represent a lower alkylene group.

R "represents a hydrogen atom, a hydroxyl group or a lower alkoxy group, R 12 and R 13 each represent a hydrogen atom or together form = 0,

Ar represents a phenyl group, a naphthyl group or a heterocyclic group, n is an integer from 1 to 3, and A represents a group> CR14 R15 or> NR14, wherein R14 represents a hydrogen atom, an optionally substituted hydroxyl group, an optionally substituted phenyl group , an acyl group, a substituted carbonyl group, an optionally substituted alkoxycarbonyl group, a substituted carbamoyl group, an optionally substituted amino group, an arylsulfonyl group, an aralkylsulfonyl group, an aralkyl group or a heterocyclic group, and R15 represents a hydrogen atom or a lower alkoxy group or R1 represents R14 together form an alkylenedioxy group of 20 or = 0, and quaternary ammonium salts of the compound of formula 1 and non-toxic salts of the compound of formula 1.

The compounds of formula 1, wherein R 2 represents a group represented by formula 3, can be prepared by performing the following steps: (1) a compound represented by formula 8 is reacted with a compound represented by formula 7, or a reactive derivative thereof or a salt thereof, wherein all the symbols in the formulas 7 and 8 have the same meanings as indicated above, and optionally carrying out one or more of the following steps (2) to (8), (2) hydrolysis for formation of a free hydroxyl group or an amino group, (3) removal of the protection by a protecting group for a hydroxyl or amino group, (4) acylating or substituted alkoxycarbonylating a hydroxyl group or an amino group, (5) alkylating a hydroxyl group or an amino group, (6) amination or hydroxylation of a carbonyl group, (7) reduction of a nitro group to an amino group, and (8) carbonylation of an acetate.

The present invention also relates to a pharmaceutical composition containing a compound of the invention.

Description of the preferred embodiments.

According to the present invention, an optionally substituted lower alkyl group comprises an unsubstituted lower alkyl group and a substituted lower alkyl group. "Lower alkyl group" means an alkyl group containing up to seven carbon atoms, preferably up to four carbon atoms. The unsubstituted lower alkyl group may have an unbranched carbon chain or a branched carbon chain and is, for example, a methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl, hexyl or heptyl group.

In the definition of R1, the substituted lower alkyl groups include an optionally substituted lower aminoalkyl group such as a 2-aminoethyl group and a 3-aminopropyl group, an N, N-dimethylaminopropyl group, lower 4-piperydylalkyl groups such as a 4-piperidylpropyl group, lower morpholinoalkyl groups such as a morpholinoethyl group and lower piperidinoalkyl such as a piperidinoethyl group. The halophenylpropargyl group includes fluorine, chlorine, bromine and iodophenylpropargyl compounds 50 and is preferably a p-chlorophenylpropargyl group. The optionally substituted aralkyl groups include unsubstituted aralkyl groups such as lower phenylalkyl groups, particularly a benzyl and phenylethyl group and substituted lower phenylalkyl groups such as a p-methoxybenzyl group. The optionally substituted phenyl group includes a substituted phenyl group such as a 3,4-dimethoxyphenyl group. The alkylene group formed by R is, for example, a methylene, ethylene, or propylene group.

A heterocyclic group Ar is, for example, a pyridyl group such as a 2-pyridyl, 3-pyridyl or 4-pyridyl group, a pyrrolyl group such as a 2-pyrrolyl or 3-pyrrolyl group, a thionyl group such as a 2-thionyl group, a 3-thionyl group or a furyl group such as a 2-furyl or 3-furyl group.

3 194549

The optionally substituted amino group R10 comprises a free amino group and a substituted amino group. In the substituted amino group, the substituents are, for example, a lower alkyl group such as a methyl, ethyl or propyl group or other lower alkyl groups as indicated above and substituted sulfo groups such as an isoquinoline sulfo, naphthalene sulfo, methanesulfo or toluene sulfo group. Thus, the substituted amino group is, for example, an isoquinolinesulfonamide group, a lower N-alkyl isoquinolinesulfonamide group such as an IM-methylsulfonamide group, a naphthalene sulfonamide group, a lower N-alkylnaphthalenesulfonamide group, a N-methylnaphthalenesulfonamide group, a methyl group, and a methane-sulfonamine group. , a toluene sulfonamide group. The substituted amino group can also be a phthalimide group.

The substituted hydroxy group R10 comprises an ester, an ether and a protected hydroxy group. The ester group is, for example, a substituted sulfonyloxy group such as an isoquinoline sulfonyloxy group, a toluene sulfonyloxy or a naphthalene sulfonyloxy group or a lower alkanoyloxy group such as an acetoxy, propionyloxy or butanoyloxy group. The ether is, for example, a lower alkoxy ether such as a methoxy, ethoxy, or propoxy ether; an aralkyloxy ether such as a benzyloxy ether; a lower alkanoyloxy-lower alkoxy ether such as an acetoxymethoxy ether; or a heterocyclic lower alkoxy such as a 2-pyridylmethoxy or 4-pyridylmethoxy ether.

The lower alkyl group R10 and the halogen atom R10 are defined as indicated above. The lower alkoxy group R10 has the same meaning as Rs, where Rs is a hydrogen atom, a halogen atom, a nitro group, a lower alkyl group, an optionally substituted hydroxyl group, an optionally N-substituted amino group, an optionally substituted carboxyl group, a polyfluoro-lower alkyl group , a cyano group, a hydroxymethyl group, a methylthio group, methylsulfinyl group or methylsulfonyl group. The halogen atom R10 is defined as indicated above for R5. The heterocyclic group Ar 2 is, for example, an imidazolyl group such as a 4-lmidazolyl group.

The substituted hydroxyl group R14 is for example an ether group such as a lower alkoxy group, defined as indicated above, or an optionally substituted aralkyl group, for example a phenyl-lower alkyl group, optionally substituted on the phenyl group, such as a benzyl, phenylpropyl, 4-methylbenzyl- or a 3,4-dichlorobenzyl group or an ester group, for example, a lower alkanoyloxy group such as an acetoxy or propanoyloxy group.

The substituted phenyl group R14 is, for example, a lower alkylphenyl group such as a 3-methylphenyl group, a lower alkoxyphenyl group such as a 2,3-dimethoxy or a 4-methoxyphenyl group, or a mono- or di-halophenyl group such as a 4-chlorophenyl or 3, 4-dichlorophenyl group.

The acyl group R14 is, for example, an acylphenyl group such as a benzoyl or an aralkylcarbonyl group such as a benzylcarbonyl or phenylpropylcarbonyl group.

The substituted alkoxycarbonyl group R14 is, for example, a phenyl-lower alkoxycarbonyl group such as a benzyloxycarbonyl group or a tert-butoxycarbonyl group.

The substituted alkoxycarbonyl group R14 is, for example, a phenyl-lower alkoxycarbonyl group such as a benzyloxycarbonyl or tert-butoxycarbonyl group.

The substituted carbonyl group is, for example, an arylcarbonyl group such as a phenylcarbonyl group or an aralkylcarbonyl group such as a benzylcarbonyl group.

The substituted amino group R14 is, for example, a lower alkylamino group, an optionally substituted aralkylamine group or Ν, Ν-lower alkylaralkylamino group, for example a methylaminobenzylamino, a 3,4-dichlorobenzylamino, a Ν, Ν-methylbenzylamino or an N, N-dimethyl-3 , 4-dichloroamino group.

The arylsulfonyl group R14 is, for example, a benzylsulfonyl or an isoquinoline sulfonyl group.

The aralkylsulfonyl group R14 is, for example, a benzylsulfonyl or a phenylpropylsulfonyl group.

The aralkyl group R14 is, for example, a benzyl or phenylpropionyl group.

The heterocyclic group R 14 is, for example, a pyridyl group such as a 2-pyridyl group or a pyrimidyl group such as a 2-pyrimidyl group.

Because the present compounds have a nitrogen atom, they can form quaternary ammonium salts or salts such as non-toxic salts. To form a quaternary ammonium salt, a compound of the present invention is reacted with, for example, methyl iodide. The salts of the present invention are non-toxic salts such as salts with an inorganic acid such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrobromic acid or hydroiodic acid, as well as salts with an organic acid such as citric acid, acetic acid, oxalic acid, tartaric acid, sulfonic acids, such as methanesulfonic acid , ethanesulfonic acid, benzenesulfonic acid, fumaric acid, maleic acid or malic acid.

According to another embodiment for the preparation of the present compound of formula 1 194549 4, a compound of formula 8 is reacted with a compound of formula 7.

The starting material of formula 8 can be obtained by reacting a compound represented by formula 10 on the formula sheet, wherein R 17 represents a hydrogen atom or a lower alkyl group with a compound represented by formula 11.

Through this reaction the compound of formula 8 is obtained wherein R12 and R13 together form = 0. A reduction of this compound gives the compound of formula 8, wherein both R12 and R13 represent a hydrogen atom.

According to a particular embodiment, a known compound tyrosine with an amino group protected and represented by formula 10a is reacted with piperazine with the protected nitrogen atom as represented by formula 11a on the formula sheet, to obtain an intermediate product represented by formula 8a on the formula sheet.

The intermediate of formula 8a is then condensed with isoquinoline sulfonyl chloride to obtain a compound represented by formula 12 in the formula sheet.

Subsequently, the following modification of the compound of formula 12 is carried out to obtain some of the present compounds: (2a) hydrolysis to remove the isoquinoline sulfonyl group to a free hydroxyl group on the phenyl ring; (3a) removal of the piperazine ring protection; (4a) acetylation or alkylation of the free hydroxyl group; (4b) acetylation of the nitrogen atom of the piperazine unit; 20 (5a) alkylation of the sulfonamide group.

To prepare other compounds of the present invention, bistidine or phenylalanine may be used instead of tyrosine and / or piperidine may be used instead of piperazine. In addition, an N * alkylated compound can be used instead of an N * protected compound 11a and / or a hydroxy-protected compound of the compound 10a can be used. The compound 8a or 12 can be reduced to convert the carbonyl structure to the methylene chain. The other part of the phenyl ring can be connected to an amino group via an alkylene chain to form a ring structure. When piperidine is used instead of piperazine, the piperidine unit can be converted to a corresponding unit with an acetal structure at its fourth position. After condensing with a sulfonic acid derivative, (8a) the acetal can be carbonylated, (6a) the carbonyl group can be converted to a hydroxy group or an amino group, (4b) the hydroxyl or amino group can be acinated, or (5b) the hydroxyl or amino group can be alkylated to obtain some of the desired compounds of the present invention.

The reaction of compounds 10 and 11 is carried out in a mixture such as tetrahydrofuran, dioxane, dichloromethane or other aprotic solvents at a temperature of about 0 ° C to 40 ° C, preferably 20 ° C to 30 ° C.

The reaction of the compound of formula 7 and the compound of formula 8 is carried out in an aprotic solvent, for example tetrahydrofuran, methylene chloride, chloroform or dimethylformamide in the presence of a base such as for example triethylamine at a temperature of about 0 ° C to 40 ° C , preferably 20 ° C to 30 ° C.

The hydrolysis of step (2) is carried out in a solvent such as, for example, methanol, tetrahydrofuran, a mixture thereof or dimethyl sulfoxide in the presence of a base such as, for example, sodium hydroxide, or potassium hydroxide.

The removal of the protection of step (3) is carried out in a solvent such as, for example, methanol, ethanol, chloroform or ethyl acetate.

The acetylation of step (4) is carried out in a solvent such as, for example, chloroform, tetrahydrofuran or pyridine in the presence of a base such as triethylamine.

The alkylation of step (5) is carried out in a solvent such as dimethylformamide, tetrahydrofuran, ethyl acetate, methanol, methylene chloride or a mixture thereof.

The hydroxylation of step (6) is carried out in a protonic solvent such as methanol or ethanol in the presence of a reducing agent such as sodium borohydride or sodium cyanoborium hydride. The amination of step (6) is carried out after the imine formation under the same conditions as before the hydroxylation. The reduction of the nitro group in step (7) is carried out in a solvent, for example an alcohol such as methanol or ethanol, by catalytic hydrogenation using a catalyst such as a noble metal catalyst such as palladium on carbon.

The conversion of acetal to oxo is carried out by acid hydrolysis in an aqueous solution. The 5 194549

Examples 1-CXXXXI and Reference Examples 1-38 belong to the present invention while Examples CXXXII-CCLXIII and Reference Examples 39-44 belong to Dutch patent 193726 which has been split off from the present invention.

5 Examples

The present invention will be further illustrated by the following examples.

In the examples, the melting points were determined by means of equipment for determining the melting point such as Vamato MP-21 (Yamato Kagaku, Japan) using a capillary; nuclear magnetic resonance spectra (1 H-NMR) were determined by JEOL.NNM-FX200 (Nippon Denshi, Japan); 10 molecular weights were determined by JMS-D300 type mass spectrometer (Nippon Denshi, Japan) and infrared absorption spectra (IR) were determined with IRA-1 (Nippon Bunko Kogyo, Japan). The results of all these determinations are stated in the original Dutch patent application 8903143. The meanings of the substituents of the compounds in the examples are listed in Table I.

Reference Example 1 1- [N- (Benzyloxycarbonyl) histidyl] -4-phenylpiperazine.

7.13 g of N, N'-dibenzyloxycarbonylhistidine, 3.00 g of 4-phenylpiperazine and 16.1 g of N-hydroxybenzotriazole were dissolved in 100 ml of tetrahydrofuran and 3.84 g of DCC (dicyclohexylcarbodiimide) were added to the mixture and the mixture was added stirred at room temperature for 3 hours. An insoluble material was filtered off, the filtrate was evaporated under reduced pressure and 200 ml of ethyl acetate was added to the concentrate to transform the crystals, which were then filtered off. The filtrate was then washed with 20% potassium carbonate in the form of an aqueous solution and a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated under reduced pressure. The resulting residue was dissolved in 60 ml of methanol and after an addition of a 10% ammonium methanol solution and stirring at room temperature for 30 minutes, the solution was evaporated under reduced pressure to obtain a residue, which was then subjected to silica gel chromatography and eluted with chloroform / methanol (50: 1 to 10: 1) to obtain 6.61 g of the title compound in a colorless, amorphous form.

Reference Example 2 1 - [N - (T ert-butoxycarbonyl) histidyl] -4-phenylpiperazine.

In 80 ml of methanol was dissolved 6.61 g of 1- [N- (benzyloxycarbonyl) histidyl] 4-phenylpiperazine and 4 g of 5% palladium on carbon as a catalyst was added to the solution while cooling with ice and the mixture was stirred under a hydrogen atmosphere filtered at room temperature for 20 hours to obtain a filtrate which was then evaporated under reduced pressure to obtain 4.26 g of a residue. The residue was dissolved in 80 ml of dimethylformamide and to the solution was then added 6.8 g of tert-butoxycarboxylic acid anhydride and 10 ml of triethylamine and the mixture was stirred at room temperature for 90 minutes. To the reaction mixture was added 200 ml of ethyl acetate which was then washed twice with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and filtered to obtain a filtrate. The filtrate was evaporated under reduced pressure to obtain a residue which was then dissolved in 100 ml of methanol and to the resulting solution was added 20 ml of 10% aqueous sodium hydroxide and the whole was stirred at room temperature for 30 minutes. The reaction mixture was evaporated under reduced pressure to one third of the original volume and after the addition of 150 ml of water, the concentrate was extracted twice with 80 ml of chloroform in each case, after which the resulting chloroform phase was dried over magnesium sulfate, filtered and evaporated under a reduced pressure to obtain a residue. The residue was applied to a silica gel column and eluted with chloroform / methanol (50: 1 to 20: 1) to obtain 4.53 g of the title compound in a colorless, amorphous form.

50

Reference Example 3 1- [2- (Tert-butoxycarbonylamino) -3-imidazole-4 (5) -yl-propyl] 4-phenylpiperazine.

A solution of 1.3 g of lithium aluminum hydride in 38 ml of tetrahydrofuran was added to a solution of 4.56 g of aluminum chloride in 38 ml of ethyl ether under ice-cooling and the mixture was stirred for 55 minutes with ice-cooling. To the mixture was added dropwise a solution of 4.53 g of 1- [N- (tert-butoxycarbonyl) histidyl] -4-phenylpiperazine in 51 ml of tetrahydrofuran and stirred for one hour while cooling with ice, after which 20 ml of 25% potassium was added to the reaction mixture 194549 6 carbonate as an aqueous solution was added, followed by 100 ml of chloroform to obtain a suspension. The slurry was filtered off using silica as a filter aid to obtain a filtrate. After the silica was washed with 20% methanol in chloroform, the combined filtrate was evaporated under reduced pressure to obtain a residue.

The residue was applied to a silica gel column and eluted with chloroform / methanol (40: 1 to 10: 1) to obtain 3.1 g of the title compound in the form of a colorless, amorphous substance.

EXAMPLE 1

N- {1- [1- (5-isoquinolinesulfonyl) imidazole-4 (5) -yl-methyl] 2- (4-phenylpiperazinyl) ethyl} -5-isoquinolinesulfonamide.

3.1 g of amorphous compound obtained in Reference Example 3 were dissolved in 10 ml of ethyl acetate and 16 ml of 4N hydrochloric acid in ethyl acetate was added to the solution and the mixture was stirred at room temperature for 30 hours and evaporated to dryness under reduced pressure. To the residue, 70 ml of tetrahydrofuran and 30 ml of chloroform were added to form a suspension to which 6 g of isoquinoline sulfonic acid chloride and 30 ml of triethylamine were added and after stirring at room temperature for 18 hours, 150 ml of water was added and the whole was extracted twice with 70 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure to obtain a residue which was then applied to a silica gel column and eluted with chloroform / methanol (80: 1 to 60: 1) to obtain 1.86 g of the in the preamble in a colorless, amorphous form.

EXAMPLE II

N- [1- (1-imidazole-4 (5) -yl-methyl) -2- (4-phenyl-piperazinyl) ethyl] -isoquinolinesulfonamide.

In a mixture of 1 ml of tetrahydrofuran and 5 ml of methanol, 250 mg of amorphous compound obtained in Example I was dissolved and 1 ml of 4N sodium hydroxide was added to the solution. After stirring at room temperature for 10 minutes, 20 ml of water was added to the mixture, which was then extracted twice with a mixture of 10 ml of chloroform and 2 ml of isopropanol. The extract was dried over magnesium sulfate and evaporated under reduced pressure to obtain a residue which was then applied to a column of silica gel and eluted with chloroform / methanol (20: 1) and chloroform / methanol / triethylamine (20: 1: 0, 2) to obtain 163 mg of the title compound in a colorless, amorphous form.

EXAMPLE III

N- {1 - [1 - (5-isoquinolinesulfonyl) imidazole-4 (5) -yl-methyl] 2- (phenylpiperazinyl) ethyl} -N-methyl-5-isoquinolinesulfonamide.

1.50 g of amorphous compound obtained in Example 1 were dissolved in 20 ml of dimethylformamide and 120 mg of 60% sodium hydride and 0.2 ml of methyl iodide were successively added to the solution while cooling with ice and after stirring for 30 minutes while cooling with ice. 30 ml of water added. After extraction of the reaction mixture, 30 ml of ethyl acetate was added, the extract was washed with an aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated under reduced pressure to obtain a residue, which was then applied to a column of silica gel and eluted with chloroform. methanol (80: 1) to obtain 616 mg of the title compound in a colorless, amorphous form.

45

EXAMPLE IV

N- [1- (1-midazol-4 (5) -yl-methyl) -2- (4-phenyl-piperazinyl) ethyl] -N-methyl-5-isoquinolinesulfonamide.

In a mixture of 2 ml of tetrahydrofuran and 10 ml of methanol, 450 mg of the amorphous compound obtained in Example III was dissolved and 1 ml of 4N sodium hydroxide was added to the solution. After stirring 50 at room temperature for 10 minutes, the reaction mixture was worked up by the same method as described in Example II to obtain 299 mg of the title compound in a colorless, amorphous form.

Reference Example 4 55 N- (Tert-butoxycarbonyl) -3,4-dibenzyloxyphenylalanine benzyl ester.

In 200 ml of dimethylformamide 21.12 g of N- (tert-butoxycarbonyl) DOPA were dissolved and after 50 g of benzyl bromide and 40 g of potassium carbonate were added, the mixture was stirred at room temperature 7 194549 for 40 hours. After the addition of 400 ml of sodium chloride dissolved in water, the reaction mixture was extracted with 500 ml of ethyl acetate and the extract was washed twice with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate, filtered and evaporated under reduced pressure. Hexane was added to the resulting residue to crystallize the title compound, which was then washed, filtered and dried to give 30.0 g of colorless crystals.

Reference Example 5 N- (Tert-butoxycarbonyl) -3,4-dibenzyloxyphenylalanine.

30.0 g of crystals obtained in Example IV were dissolved in 600 ml of methanol and after the addition of 65 ml of 10% sodium hydroxide, the mixture was stirred at room temperature for 20 minutes and 1000 ml of water was added. The reaction mixture was adjusted to pH = 4 with concentrated hydrochloric acid and extracted twice with 800 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure to crystallize the title compound, which was then filtered and washed with hexane to give 25.2 g of colorless crystals.

Reference Example 6 1- [N- (Tert-butoxycarbonyl) -3,4-dibenzyloxyphenylalaninyl] 4-phenylpiperazine.

In 80 ml of methylene chloride, 5.67 g of crystals obtained in Reference Example were dissolved, 5.9 g of N-phenylpiperazine and 1.53 g of N-hydroxybenzotriazole, and after the addition of 2.4 g of DCC, the mixture was stirred at room temperature for 18 hours. The resulting insoluble material was filtered off and washed with ethyl acetate.

The combined filtrate was evaporated under reduced pressure to obtain a residue which was then applied to a silica gel column and eluted with hexane / ethyl acetate (2: 1) to obtain 6.39 g of the title compound in the title. form of a colorless, amorphous material.

Reference Example 7 1- {2- [N - (Tert-butoxycarbonylamino] -3- (3,4-dibenzyloxyphenyl) propyl} -4-phenylpiperazine.

In 50 ml of tetrahydrofuran, 3.66 g of colorless, amorphous material obtained in Reference Example 6 was dissolved and the addition of 700 mg of lithium aluminum hydride while cooling with ice was carried out, the mixture being stirred for 90 minutes while cooling with ice and to the mixture. water was added until the foam formation was terminated. Then 80 ml of chloroform was added to the reaction mixture to form a suspension which was then filtered using silica gel 35 as a filter aid, to remove the insoluble material. The resulting filtrate was evaporated under reduced pressure to obtain a residue which was then applied to a silica gel column and eluted with hexane / ethyl acetate (3: 1) to obtain 2.67 g of the title compound in the colorless , amorphous form.

Reference Example 8 1- [2-Amino-3- (3,4-dibenzyloxyphenyl) propyl] -4-phenylpiperazine.

4.35 g of amorphous compound obtained in Reference Example 7 was dissolved in 20 ml of ethyl acetate and after the addition of 30 ml of 4N hydrochloric acid in ethyl acetate, the mixture was stirred at room temperature for one hour. The reaction mixture was evaporated under reduced pressure, made alkaline with aqueous sodium bicarbonate 45 and extracted twice with 80 ml of chloroform and the extract was dried over magnesium sulfate and evaporated under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (100: 1 to 30: 1) to obtain 1.64 g of the title compound in a colorless, amorphous form.

Example V

N- {1 - [(3,4-Dibenzyloxyphenyl) methyl] -2- (4-phenylpiperazinyl) ethyl} 5-isoquinoline sulfonamide.

In 15 ml of methylene chloride, 640 mg of amorphous compound obtained in Reference Example 8 was dissolved and 1 ml of triethylamine and 350 mg of 5-isoquinolinesulfonyl chloride was added to the solution while cooling with ice and after stirring for 1 hour while cooling with ice was added 50 ml of water and the 55 mixture was extracted twice with 50 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure to obtain a residue which was then applied to a column of silica gel and eluted with hexane / ethyl acetate (1: 1) to obtain 470 mg of the compound mentioned in the 194549 8 heading. in a colorless, amorphous form.

Example VI

N- {1 - [(3,4-Dibenzyloxyphenyl) methyl] -2- (4-phenylpiperazinyl) ethyl} -N-methyl-5-isoquinolinesulfonamide.

470 mg of amorphous compound obtained in Example 5 were dissolved in 8 ml of dimethylformamide and 30 mg of 60% sodium hydride and 0.1 ml of methyl iodide were successively added to the solution while cooling with ice and after stirring for 2 hours while cooling with ice, a saturated aqueous sodium chloride solution was added and the mixture was extracted with 50 ml of ethyl acetate. The extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated under reduced pressure to give a residue which was then applied to a silica gel column and eluted with hexane / ethyl acetate (1: 1) to obtain 413 mg of the title compound in a colorless, amorphous form.

EXAMPLE VII

N- {1 - [(3,4-Dihydroxyphenyl) methyl] -2- (4-phenylpiperazinyl) ethyl} -N-methyl isoquinolinesulfonamide.

310 mg of amorphous compound obtained in Example 6 were dissolved in 2 ml of 1,2-ethanedithiol and 1 ml of boron trifluoride / ethyl ether was added to the solution and after stirring at room temperature for 18 hours a saturated aqueous solution of sodium bicarbonate was added and the reaction mixture was added two times. extracted once with a mixture of chloroform and methanol (10: 1). The extract was dried over magnesium sulfate and evaporated under reduced pressure to obtain a residue which was then applied to a silica gel column and eluted with chloroform / methanol (80: 1 to 20: 1) to obtain 148 mg of the the above compound in a colorless, amorphous form.

EXAMPLE VIII

N- {1 - [(3,4-Dihydroxyphenyl) methyl] -2- (4-phenylpiperazinyl) ethyl} -5-isoquinolinesulfonamide.

The amorphous compound obtained in Example V was treated according to the method described in Example VII to obtain the title compound in a colorless, amorphous form.

Reference Example 9 6,7-Dibenzyloxy-3 - [(4-phenylpiperazinyl) methyl] -1,2,3,4-tetrahydroisoquinoline.

1.00 g of amorphous compound obtained in Reference Example 8 were dissolved in 2 ml of tetrahydrofuran and 0.25 ml of 37% formalin was added to the solution. After stirring at room temperature for 30 minutes, 600 mg of 12 N hydrochloric acid was added to the mixture, which was then stirred at room temperature for two hours. After the addition of a saturated aqueous solution of sodium bicarbonate, the reaction mixture was extracted twice with 20 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure to obtain a residue which was then applied to a silica gel column and eluted with chloroform / methanol (100: 1) to obtain 585 mg of the title compound in a colorless, amorphous form.

Example IX

6.7-Dibenzyloxy-2- (5-isoquinolinesulfonyl) -3 - [(4-phenylpiperazinyl) methyl] -1,2,3,4-tetrahydroisoquinoline.

580 mg of the amorphous compound obtained in Reference Example 9 was dissolved in 10 ml of methylene chloride and 1 ml of triethylamine and 400 mg of 5-isoquinolinesulfonyl chloride.HCl were added to the solution while cooling with ice. The mixture was stirred at room temperature for two hours and after the addition of 20 ml of water, it was extracted twice with 10 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure to obtain a residue which was then applied to a silica gel column and eluted with hexane / ethyl acetate (1: 1) to obtain 610 mg of the title compound in a colorless, amorphous form.

Example X

6.7-Dihydroxy-2- (5-isoquinolinesulfonyl) -3 - [(4-phenylpiperazinyl) methyl] -1,2,3,4-tetrahydroisoquinoline.

To 314 mg of amorphous compound obtained in Example IX was added 2 ml of 1,2-ethanedithiol and 1 ml of boron trifluoride / ethyl ether and the mixture was stirred at room temperature for 18 hours. After the addition of a saturated aqueous solution of sodium bicarbonate, the reaction mixture was extracted twice with a mixture of chloroform and methanol (1: 1) and the extract was dried over magnesium sulfate and evaporated under reduced pressure to obtain a residue which was then was applied to a silica gel column and eluted with chloroform / methanol (50: 1 to 20: 1) 9 194549 to obtain 213 mg of the title compound in the colorless, amorphous form. Reference Example 10 1- [N- (tert-butoxycarbonyl) -β-nitrophenylalanyl] -4-phenylpiperazine.

In 70 ml of 1,4-dioxane, 7.03 g of p-nitrophenylalanine was suspended and 28 ml of 10% sodium hydroxide in the form of an aqueous solution and 7.5 g of di-tert-butyl dicarbonate were added to the suspension and the mixture was added. stirred at room temperature for 30 minutes. To the reaction mixture, 200 ml of water and 7 ml of 12 N hydrochloric acid were added, which was then extracted with 150 ml of ethyl acetate and the extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated under reduced pressure. The resulting residue was dissolved in 150 ml of tetrahydrofuran and to the solution was added 6.0 g of N-phenylpiperazine and 5.5 g of N-hydroxybenzotriazole and further 7.6 g of DCC were added. After stirring at room temperature for 3 hours, the reaction mixture was filtered to remove insoluble material and the filtrate was evaporated under reduced pressure and the resulting residue was dissolved in 200 ml of ethyl acetate. The solution was successively washed with 10% aqueous solution of potassium carbonate and saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated under reduced pressure to obtain a residue which was then applied to a silica gel column and eluted with hexane / ethyl acetate (2: 1) to obtain 11.1 g of the title compound in the form of light yellow crystals.

Example XI

1- [N- (5-Isoquinolinesulfonyl) -p-nitrophenylalanyl] -4-phenylpiperazine.

In 100 ml of ethyl acetate, 11.0 g of crystals obtained in Reference Example 10 were dissolved and after the addition of 100 ml of 4N hydrochloric acid in ethyl acetate, the reaction mixture was stirred at room temperature for one hour and evaporated to dryness under reduced pressure. To the residue, 200 ml of a saturated aqueous solution of sodium bicarbonate was added and the mixture was extracted twice with 100 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure to give a residue which was then applied to a silica gel column and eluted with chloroform / methanol (80: 1 to 10: 1) to give a free amine. The free amine was dissolved in 100 ml of methylene chloride and 8.5 g of 5-isoquinolinesulfonyl chloride.HCl and 20 ml of triethylamine were successively added to the solution. The reaction mixture was stirred at room temperature for 18 hours and after the addition of water extracted twice with 100 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure to give a residue which was then applied to a silica gel column and eluted with chloroform / methanol (100: 1 to 50: 1) to obtain 9.66 g of the in the above compound in the form of colorless crystals.

Example XII

1- [N- (5-Isoquinolinesulfonyl) -N-methyl-p-nitrophenylalanyl] 4-phenylpiperazine.

In 60 ml of dimethylformamide, 5.87 g of crystals obtained in Example XI were dissolved and to the 40 solution were added successively 500 mg of 60% sodium hydride and 1.5 ml of methyl iodide while cooling with ice. After stirring for two hours while cooling with ice, water was added to the reaction mixture, which was then extracted with 150 ml of ethyl acetate. The extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated under reduced pressure to give a residue applied to a column of silica gel eluted 45 with chloroform / methanol (100: 1) to obtain 5, 93 g of the compound mentioned in the preamble in a yellow amorphous form.

EXAMPLE XIII

1- [p-Amino-N- (5-isoquinolinesulfonyl) -N-methylalanyl] -4-phenylpiperazine.

50.08 g of amorphous compound obtained in Example XII were dissolved in 70 ml of methanol and 5 ml of 12 N hydrochloric acid and 30 ml of water were added to the solution and then 5 g of 5% palladium on carbon. The mixture was stirred at room temperature under hydrogen atmosphere for 30 minutes and filtered to remove the insoluble material and the filtrate was evaporated under reduced pressure and to the residue was added 150 ml of saturated aqueous solution of sodium bicarbonate and the 55 mixture was extracted twice with 200 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure to obtain a residue which was then applied to a column of silica gel and eluted with chloroform / methanol (50: 1) to obtain 3.32 g of the solution in 194549 The above compound in a yellow, amorphous form.

Example XIV

According to the method described in Example XIII, 3.75 g of crystals prepared in Example XI were treated to obtain 2.17 g of 1- [p-amino-N- (5-isoquinolinesulfonyl) phenylalanyl] -4-phenylpiper azine in the form of yellow crystals.

Example XV

1- [N- (5-Isoquinolinesulfonyl) -p- (p-toluenesulfonylamino) phenylalanyl] -4-phenylpiperazine.

In 5 ml of pyridine, 200 mg of the crystals obtained in Example XIV were dissolved and to the solution was added 90 mg of p-toluenesulfonyl chloride while cooling with ice and the mixture was stirred for 1 hour while cooling with ice and poured over 30 ml of saturated aqueous sodium bicarbonate solution. The mixture was extracted twice with 15 ml of chloroform and the extract was dried over magnesium sulfate and evaporated under reduced pressure to obtain a residue which was then applied to a silica gel column and eluted with chloroform / methanol (80: 1 to 50: 1) to obtain 128 mg of the compound mentioned in the preamble.

Example XVI

The same procedure as described in Example XV was repeated except that 200 mg of 5-isoquinolinesulfonyl chloride.HCl and 300 mg of crystals obtained in Example XIV were used and the elution was carried out with chloroform / methanol (40: 1 to 20: 1) to obtain of 372 mg of 1- [N - (5-isoquinolinesulfonyl) -p- (5-isoquinolinesulfonylamino) phenylalanyl] -4-phenylpiperazine.

Example XVII

The same procedure as described in Example XV was repeated, except that 200 mg of 1-naphthalenesulfonyl chloride and 360 mg of crystals obtained in Example XIV were used as starting materials and the elution was carried out using chloroform / methanol (80: 1 to 50: 1) to obtain 385 mg of 1- [N- (5-isoquinolinesulfonyl) p- (1-naphthalenesulfonylamino) phenylalanyl] -4-phenylpiperazine.

30

Example XVIII

The same procedure as described in Example XV was repeated except that 0.07 ml of methanesulfonyl chloride and 360 mg of the crystals obtained in Example XIV were used as starting materials and the elution was performed using chloroform / methanol (50: 1 to 30 : 1) to obtain 35 356 mg of 1- [N - (5-isoquinolinesulfonyl) -p- (methanesulfonylamino) phenylalanyl] -4-phenylpiperazine.

Example XIX

1- [N- (5-Isoquinolinesulfonyl) -p-methanesulfonylamino-N-methylphenylalanyl] -4-phenylpiperazine.

In 7 ml of pyridine, 700 mg of amorphous compound obtained in Example XIII were dissolved and to the solution was added 0.13 ml of methanesulfonyl chloride while cooling with ice and the mixture was stirred for 1 hour while cooling with ice and poured into 50 ml of saturated aqueous solution of sodium bicarbonate. The mixture was extracted twice with 30 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (100: 1 to 50: 1) to obtain 45 790 mg of the title compound.

Example XX

The same procedure as described in Example XIX was repeated except that 360 mg of 1-naphthalenesulfonyl chloride and 700 mg of amorphous compound obtained in Example XIII were used as 50 starting materials and the elution was performed using chloroform / methanol (100: 1) to obtain 770 mg of 1- [N- (5-isoquinolinesulfonyl) -N-methyl-p- (1-naphthalenesulfonylamino) phenylalanyl] -4-phenylpiperazine.

Example XXI

The same procedure as described in Example XIX was repeated except that 320 mg of 5-isoquinoline sulfonyl chloride.HCl was used as sulfonating agent, 500 mg of amorphous compound obtained in Example XIII, 5 ml of pyridine and chloroform / methanol (80: 1 to 50: 1). 11454949 were used as eluant to obtain 498 mg of 1- [N- (5-isoquinolinesulfonyl) -p- (5-isoquinolinesulfonylamino) -N-methylphenylalanyl] 4-phenylpiperazine.

Example XXII

The same procedure as described in Example XIX was repeated except that 300 mg of p-toluenesulfonyl chloride as sulfonating agent, 700 mg of amorphous compound obtained in Example XIII, 10 ml of pyridine as eluate and chloroform / methanol (100: 1) were used to obtain 812 mg of 1- [N- (5-isoquinolinesulfonyl) -N-methyl-p- (p-toluenesulfonylamino) phenylalanyl] -4-phenylpiperazine.

Example XXIII

1- {N- (5-Isoquinolinesulfonyl) -p- [N '- (5-isoquinolinesulfonyl) -N'-methylamino] -N-methylphenylalanyl} -4-phenylpiperazine.

In 5 ml of dimethylformamide, 306 mg of the product in Example XXI was dissolved and to the solution were added 25 mg of 60% sodium hydride and 0.1 ml of hydrogen iodide while cooling with ice, and the mixture was stirred for 1 hour while cooling with ice. After the addition of 30 ml of saturated aqueous solution of sodium chloride, the mixture was extracted with 30 ml of ethyl acetate and the extract was washed with an aqueous, saturated solution of sodium chloride, dried over magnesium sulfate and evaporated under reduced pressure. The resulting residue was applied to a column of silica gel and eluted with chloroform / methanol (80: 1) to obtain 266 mg of the title compound.

Example XXIV

The same process as described in Example XXIII was repeated, except that 594 mg of the product obtained in Example XIX was dissolved in 6 ml of dimethylformamide and to the solution was added 60 mg of 60% sodium hydride and 0.1 ml of methyl iodide to obtain 450 mg 1- [N- (5-isoquinolinesulfonyl) -p- (N'-methanesulfonyl-N-methylamino) -N-methylphenylalanyl] -4-phenylpiperazine.

Example XXV

The same process as described in Example XXIII was repeated except that 587 mg of the product of Example XX was dissolved in 6 ml of dimethylformamide and to the solution was added 50 mg of 60% sodium hydride and 0.1 ml of methyl iodide and the elution was carried out under use of chloroform / methanol (100: 1) to obtain 490 mg of 1- {N- (5-isoquinolinesulfonyl) -N-methyl-p- [N'-methyl-N- (1-naphthalenesulfonyl) amino] phenylalanyl} - 4-phenylpiperazine.

Example XXVI

The same procedure as described in Example XXIII was repeated except that 650 mg of the product obtained in Example XXII was dissolved in 10 ml of dimethylformamide and to the solution was added 60 mg of 60% sodium hydride and 0.1 ml of methyl iodide and the elution was carried out under use of chloroform / methanol (100: 1) to obtain 603 mg of 1- {N- (5-isoquinolinesulfonyl) -N-methyl-p-40 [N'-methyl-N- (p-toluenesulfonyl) amino] phenylalanyl} - 4-phenylpiperazine.

Reference Example 11 1- (N-Benzyloxycarbonyltyrosyl) -4- (tert-butoxycarbonyl) piperazine.

In a mixed solvent of 200 ml of methylene chloride and 100 ml of ethyl acetate, 21.31 g of 45 N-benzyloxycarbonyl tyrosine and 11.79 g of N- (tert-butoxycarbonyl) piperazine were dissolved and 14 g of DCC was added to the solution. After stirring at room temperature for 40 hours, insoluble material precipitated which was filtered off and the filtrate was evaporated under reduced pressure and the resulting residue was applied to a silica gel column and eluted with hexane / ethyl acetate (1: 1) to obtain 23.9 g of the compound mentioned in the preamble in a colorless, amorphous form.

50

Example XXVII

1- [N, O-bis (5-isoquinolinesulfonyl) tyrosyl] -4- (tert-butoxycarbonyl) piperazine.

1.00 g of amorphous compound obtained in Reference Example 11 were dissolved in 20 ml of methanol, 500 mg of 5% palladium on carbon were added to the solution and the mixture was stirred under a hydrogen atmosphere at room temperature for 5 hours. After removal of the insoluble material by filtration, the filtrate was evaporated under reduced pressure. To the resulting residue were added 30 ml of tetrahydrofuran, 630 mg of 5-isoquinolinesulfonyl chloride.HCl and 1.4 ml of triethylamine consecutively and the mixture was stirred at room temperature for 50 hours and twice after the addition of 100 ml of water. extracted with 50 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure. The resulting residue was then applied to a silica gel column and eluted with chloroform / methanol (50: 1 to 25: 1) to obtain 1.38 g of the compound mentioned in the preamble in a yellow, amorphous form.

Example XXVIII

1- [N, 0-Bis (5-isoquinolinesulfonyl) tyrosyl] piperazine.

In 3 ml of chloroform, 366 mg of amorphous compound prepared in Example XXVII was dissolved and 5 ml of 3N hydrochloric acid / ethyl acetate was added to the solution. After stirring at room temperature for one hour, the mixture was evaporated under reduced pressure and to the resulting residue was added 50 ml of saturated aqueous solution of sodium bicarbonate. The mixture was then extracted twice with 30 ml of a chloroform / methanol mixed solvent (5: 1) and the extract was dried over magnesium sulfate and evaporated under reduced pressure to obtain 301 mg of a crude preparation of the title compound. in a colorless, amorphous form.

Example XXIX

1-Benzyloxycarbonyl-4- [N - (5-isoquinolinesulfonyl) tyrosyl] piperazine.

In 10 ml of methylene chloride, 620 mg of crude product obtained in Example XXVIII was dissolved and 0.29 ml of benzyloxycarbonyl chloride and 3.04 ml of triethylamine were added to the solution in succession while cooling with ice. After stirring for 2 hours while cooling with ice, 40 ml of a saturated aqueous solution of sodium chloride was added to the reaction mixture, which was then extracted twice with 20 ml of chloroform and the extract was dried over magnesium sulfate and evaporated under reduced pressure to obtain a residue. The residue was dissolved in 6 ml of methanol and after the addition of 2 ml of 1N sodium hydroxide in the form of an aqueous solution, the mixture was refluxed for two hours, dried over magnesium sulfate and evaporated under reduced pressure. The resulting residue was applied to a column of silica gel and eluted with chloroform / methanol (80: 1 to 50: 1) to obtain 336 mg of the title compound in the form of colorless crystals.

30

Example XXX

The same process as described in Example XXIX was repeated to give 1- [N- (5-isoquinolinesulfonyl) tyrosyl] -4-phenylacetylpiperazine in a yellow, amorphous form.

Example XXXI

The same process as described in Example XXIX was repeated to give 1- [N- (5-isoquinolinesulfonyl) tyrosyl] -4- (3-phenylpropionyl) piperazine in the form of colorless crystals.

Vborbee / cf XXXII

40 1- [N, O-Bis (5-isoquinolinesulfonyl) tyrosyl] -4- (3-phenylpropyl) piperazine.

301 mg of crude product obtained in Example XXVIII and 95 mg of 3-phenylpropyl bromide were dissolved in 5 ml of dimethylformamide and 66 mg of potassium carbonate and 72 mg of sodium iodide were added to the solution. After stirring at 80 ° C for 7 hours, 30 ml of saturated aqueous sodium chloride solution was added to the reaction mixture, which was then extracted with 40 ml of ethyl acetate 45 and the extract was washed with 30 ml of saturated aqueous sodium chloride solution, dried over magnesium sulfate and evaporated under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (40: 1) to obtain 216 mg of the title compound in a yellow, amorphous form.

50 Example XXXIII

1- [N- (5-Isoquinolinesulfonyl) tyrosyl] -4- (3-phenylpropyl) piperazine.

216 mg of amorphous compound obtained in Example XXXII were dissolved in 3 ml of methanol and 0.6 ml of 2N potassium hydroxide in the form of an aqueous solution was added to the solution. The mixture was refluxed for 10 hours and after the addition of 30 ml of saturated, 55 aqueous sodium chloride solution extracted twice with 20 ml of mixed chloroform / isopropanol solvent (5: 1). The extract was dried over magnesium sulfate and evaporated under reduced pressure and a resulting residue was applied to a column of silica gel and eluted with 13 194549 chloroform / methanol (40: 1 to 10: 1) to obtain 74 mg of the title mentioned in the preamble. compound in a colorless, amorphous form.

Reference Example 12 1- [N- (Tert-butoxycarbonyl) tyrosyl] -4-phenylpiperazine.

In 100 ml of tetrahydrofuran, 19.7 g of N- (tert-butoxycarbonyl) tyrosine, 12.5 g of N-phenylpiperazine and 16.1 g of N-hydroxybenzotriazole were dissolved and a solution of 18.7 g of DCC was added dropwise to the solution. 50 ml of tetrahydrofuran for 20 minutes while cooling with ice and the mixture was stirred for one hour. The reaction mixture was filtered to remove the insoluble material which was then washed with 300 ml of ethyl acetate and the filtrate was combined and evaporated under reduced pressure. The resulting residue was dissolved in 500 ml of ethyl acetate and the solution was washed three times with a saturated aqueous solution of sodium bicarbonate and once with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated under reduced pressure. The resulting residue was applied to a silica gel column and eluted with ethyl acetate / hexane (1: 2 to 1: 1) to collect the desired fractions which were then combined and evaporated under reduced pressure. The resulting residue was dissolved in 100 ml of ethyl acetate and the solution was stored in a cooler overnight and then filtered to remove the insoluble material. The filtrate was evaporated under reduced pressure, subjected to azeotropic distillation with benzene and dried under reduced pressure to obtain 40.0 g of the title compound in a colorless, amorphous form.

Reference Example 13 1- [2- (Tert-butoxycarbonylamino) -3- (p-hydroxyphenyl) propyl] 4-phenylpiperazine.

While cooling with ice, a solution of 28.0 g of aluminum chloride in 230 ml of ether was added to a solution of 8.0 g of lithium aluminum hydride in 230 ml of tetrahydrofuran, which was added dropwise for 50 minutes and after 15 minutes the resulting solution added dropwise a solution of 40.0 g of amorphous compound obtained in Reference Example 12 in 230 ml of tetrahydrofuran for 15 minutes. The reaction mixture was brought to room temperature and after the addition of 300 ml of tetrahydrofuran stirred for 25 minutes. The mixture was filtered to remove the insoluble material which was then washed with tetrahydrofuran. The combined filtrate was evaporated under reduced pressure and the resulting residue was applied to a silica gel column and eluted with chloroform / methanol (20: 1) to collect fractions which were then evaporated under reduced pressure. Then 100 ml of ethyl acetate was added to the residue to crystallize a product. The product was filtered off to collect it and was washed five times with a mother liquor and further three times with n-hexane and dried under reduced pressure to obtain 24.1 g of the title compound in the form of colorless crystals .

Reference Example 14 1- [2-Amino-3- (p-hydroxyphenyl) propyl] -4-phenylpiperazine.

40 To a suspension of 23.6 g of the crystals obtained in Reference Example 13 in 100 ml of ethyl acetate was added dropwise 215 ml of 4N hydrochloric acid solution in ethyl acetate for 30 minutes and after stirring for 90 minutes the excess of hydrochloric acid was removed from the reaction mixture under a reduced pressure. After extraction with 200 ml of water, the separated layer of ethyl acetate was extracted with 50 ml of 1N aqueous hydrochloric acid solution. The aqueous layers were combined and neutralized to pH 7.4 with solid sodium bicarbonate and the resulting crystals were collected, washed with water and benzene, and dried by phosphorus pentaoxide in a desiccator under reduced pressure to obtain 16.9 g of the compound mentioned in the preamble in the form of colorless crystals.

50 Example XXXIV

N- {1- [p- (5-isoquinolinesulfonyloxy) benzyl-2- (4-phenylpiperazinyl) ethyl} -5-isoquinolinesulfonamide.

To a suspension of 22.96 g of crystals obtained in Reference Example 14 in 700 ml of tetrahydrofuran was added 51.01 g of 5-isoquinolinesulfonyl chloride.HCl with ice cooling for 5 minutes and then 103 ml of triethylamine was added dropwise for 30 minutes. After the reaction-55 mixture was heated to room temperature, it was poured into 460 ml of ice water and the whole was extracted with 920 ml and 230 ml of chloroform. The combined extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated to dryness under reduced pressure. The resulting amorphous residue was applied to a column of silica gel and eluted with chloroform / methanol (100: 1 to 50: 1) to obtain 45.5 g of the title compound in a yellow, amorphous form.

Example XXXV

N- {1- [p- (5-isoquinoline sulfosloxy) benzyl] -2- (4-phenylpiperazinyl) ethyl} -N-methyl-5-isoquinolinesulfonamide.

To a solution of 25.0 g of the amorphous compound obtained in Example XXXIV in 200 ml of dimethylformamide was added 3.64 g of 60% sodium hydride in three portions and after 5 minutes 3.14 ml of methyl iodide was also added dropwise for 2 minutes and the reaction mixture was stirred for 1 hour. The reaction mixture was poured onto 400 ml of ice water and the whole was extracted with 200 ml, 200 ml and 100 ml of ethyl acetate. The combined extract was washed three times with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated under reduced pressure, and the resulting residue was applied to a column of silica gel and eluted with chloroform / methanol (100: 1) to obtain 20. 0 g of the compound mentioned in the preamble in a yellow, amorphous form.

Example XXXVI

N- [1- (p-Hydroxybenzyl) -2- (4-phenylpiperazinyl) ethyl] -N-methyl-5-isoquinolinesulfonamide.

To 17.7 g of the amorphous compound obtained in Example XXXV were added 240 ml of methanol, 60 ml of tetrahydrofuran and 29 ml of 2N sodium hydroxide dissolved in water and the mixture was refluxed for 150 minutes and then poured over a saturated aqueous solution of sodium chloride. The mixture was extracted three times with 200 ml of chloroform and the extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated under reduced pressure. The resulting residue was applied to a column of silica gel and eluted with chloroform / methanol (50: 1), and 10.9 g of a yellow, amorphous product was obtained from the eluate. To the product was added 54 ml of ethanol and the mixture was stirred at room temperature for one hour and with ice cooling for 30 minutes to form crystals which were then collected, washed three times with mother liquor and twice with benzene and dried under a reduced pressure to obtain 8.2 g of the compound mentioned in the preamble in the form of light yellow crystals.

Example XXXVII

The amorphous compound obtained in Example XXXIV was subjected to an alkaline hydrolysis according to the method described in Example XXXVI to obtain N- [1- (p-hydroxybenzyl) -2- (4-35 phenylpiperazinyl) ethyl] -5-isoquinolinesulfonamide in the form of a colorless, amorphous substance.

Example XXXVIII

N- [1- (p-Methoxybenzyl) -2- (4-phenylpiperazinyl) ethyl] -N-methyl-5-isoquinolinesulfonamide.

In 20 ml of a mixed solvent of dimethylformamide / tetrahydrofuran (1: 1), 1.51 g of the 40 crystals obtained in Example XXXVI were dissolved and to the solution was added 140 mg of 60% sodium hydride with stirring and cooling with ice and the stirring was continued for about 30 minutes. After the foam formation was terminated, 490 mg of methyl iodide was added and the mixture was further stirred at room temperature overnight. After the addition of ice, the reaction mixture was extracted three times with 50 ml of ethyl acetate and the extract was washed with a 45 saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (100: 1) to obtain 1.55 g of the title compound in the form of a light brown oil.

50 Example XXXIX

The same procedures as described in Reference Examples 12 to 14 and Example XXXIV were repeated, except that 1- (2-pyrimidyl) piperazine dihydrochloride was used instead of N-phenylpiperazine to obtain N- {1 * [p- (5- isoquinolinesulfonyloxy) benzyl] -2- [4- (2-pyrimidyl) piperazinyl] ethyl} -5-isoquinoline sulfonamide in a colorless, amorphous form.

15 194549

Example XL

The amorphous compound of Example XXXIX was treated as described in Example XXXVII to give N- {1- (p-hydroxybenzyl) 2- [4- (2-pyrimidyl) piperazinyl] ethyl} -5-isoquinolinesulfonamide in a colorless, amorphous form .

5

Example XLI

The amorphous compound of Example XXXIX was treated as described in Example XXXV to obtain N- {1- [p- (5-isoquinolinesulfonyloxy) benzyl] -2- [4- (2-pyrimidyl) piperazinyl] ethyl} -N-methyl -5-isoquinolinesulfonamide in a colorless, amorphous form.

10

Example XLII

The amorphous compound of Example XLI was treated as described in Example XXXVI to obtain N- {1 - (p-hydroxybenzyl) 2- [4- (2-pyrimidyl) piperazinyl] ethyl} -N-methyl-5-isoquinolinesulfonamide in a colorless, amorphous form.

15

Example XUU

The same methods as described in Reference Examples 12 to 14 and in Examples XXXIV and XXXV were repeated, except that N- (tert-butoxycarbonyl) phenylalanine was used instead of N- (tert-butoxycarbonyl) tyrosine to obtain N- [1 - benzyl 2- (4-phenylpiperazinyl) ethyl] -N-methyl-5-20 isoquinolinesulfonamide in a light yellow amorphous form.

Example XLIV

The same procedures as described in Reference Examples 12 to 14 and Example XXXIV were repeated, except that N- (2-pyridyl) plperazine was used instead of N-phenylpiperazine to obtain N- {1- [p- (5-isoquinolinesulfonyloxy) benzyl 2- [4- (2-pyridyl) piperazinyl] ethyl} -5-isoquinolinesulfonamide in a yellow, amorphous form.

Example XLV

The product of Example XLIV was treated as described in Example XXXV to obtain 30 N- {1- [p- (5-isoquinolinesulfonyioxy) benzyl] -2- [4- (2-pyridyl) piperazinyl] ethyl} -N-methyl -5-isoquinolinesulfonamide.

Example XLVI

The product of Example XLV was treated as described in Example XXXVI to obtain 35 N- {1- (p-hydroxybenzyl) -2- [4- (2-pyridyl) piperazinyl] ethyl} -N-methyl-5-isoquinolinesulfonamide.

Example XLVII

The same methods as described in Reference Examples 12 to 14 and Examples XXXIV to XXXVI were repeated, except that N- (m-chlorophenyl) piperazine was used instead of N-phenylpiperazine to obtain N- {2- [4- (m-) chlorophenyl) piperazinyl] -1- (p-hydroxybenzyl) ethyl} -N-methyl-5-isoquinolinesulfonamide in a light yellow amorphous form.

Example XLVIII

The same procedures as described in Reference Examples 12 to 14 and Example XXXIV were repeated 45, except that N- (p-fluorophenyl) piperazine was used instead of N-phenylpiperazine to obtain N- {2- [4- (p-fluorophenyl)]. piperazinyl] -1 - [p- (5-isoquinolinesulfonyloxy) benzyl] ethyl]} - 5-isoquinoline sulfonamide in a colorless, amorphous form.

Example IL

The amorphous compound obtained in Example XLVIII was methylated according to the method described in Example XXXV to obtain N- {2- [4- (p-fluorophenyl) piperazinyl] -1- [p- (5-isoquinolinesulfonyloxy) benzyl] ethyl ]} - N-methyl-5-isoquinolinesulfonamide in a light yellow, amorphous form.

Example L

55 According to the method described in Example XXXVI, 160 mg of the amorphous compound obtained in Example IL was dissolved in 2 ml of methanol and 0.5 ml of 2N sodium hydroxide was added to the solution and the reaction mixture was refluxed for two hours, cooled and three 194549 16 extracted with chloroform. The extract was purified on a silica gel column using chloroform / methanol (100: 2) to obtain 103 mg of N- {1- (p-hydroxybenzyl) 2- [4- (p-fluorophenyl) piperazinyl] ethyl } -N-methyl-5-isoquinolinesulfonamide in a light yellow, amorphous form.

Example LI

The same procedures as described in Reference Examples 12 to 14 and Examples XXXIV to XXXVI were repeated, except that N- (m-methylphenyl) piperazine was used instead of N-phenylpiperazine to obtain N- {1 - (p-hydroxybenzyl) -2 - [4- (m-methylphenyl) piperazinyl] ethyl} -N-methyl-5-isoquinolinesulfonamide in a light yellow amorphous form.

10

Example UI

The same procedures as described in Reference Examples 12 to 14 and Example XXXIV were repeated sequentially, except that N- (p-methoxyphenyl) piperazine was used instead of N-phenylpiperazine to obtain N- {1- [p- (5-isoquinolinesulfonyloxy) benzyl 2- [4- (p-methoxyphenyl) piperazinyl] ethyl} -5-isoquinolinesulfonamide in a yellow, amorphous form.

Example III

The amorphous compound of example Lil was treated with methyl iodide according to the method described in example XXXV to obtain N- {1- [p- (5-isoquinolinesulfonyloxy) benzyl] -2- [4- (p-methoxyphenyl) piperazinyl] ethyl } -N-methyl-5-isoquinolinesulfonamide in a yellow, amorphous form.

Example LIV

The amorphous compound obtained in Example 11 was subjected to an alkaline hydrolysis according to the method described in Example XXXVI to obtain N- {1- (p-hydroxybenzyl) -2- [4- (p-25 methoxyphenyl) piperazinyl] ethyl} - N-methyl-5-isoquinolinesulfonamide in the form of yellow crystals.

Example LV

The amorphous compound obtained in example Lil was subjected to an alkaline hydrolysis according to the method described in example XXXVII to obtain N- {1- (p-hydroxybenzyl) -2- [4- (p-30 methoxyphenyl) piperazinyl] ethyl} - 5-isoquinolinesulfonamide as yellow crystals.

Example LVI

The same procedures as described in Reference Examples 12 to 14 and Examples XXXIV and XXXV were repeated sequentially, except that N- (2-methoxyphenyl) piperazine was used instead of N-phenylpiperazine to give N- {1- [p- (5) -isoquinolinesulfonyloxy) benzyl] -2- [4- (2-methoxyphenyl) piperazinyl] ethyl} -N-methyl-5-isoquinolinesulfonamide and 800 mg of the compound were subjected to an alkaline hydrolysis according to the method described in Example XXXVI to obtain 504 mg of N- {1- (p-hydroxybenzyl) -2- [4- (2-methoxyphenyl) piperazinyl] ethyl} -N-methyl-5-isoquinolinesulfonamide in a light yellow amorphous form.

40

Example LVII

1- [2-Amino-3- (p-hydroxyphenyl) propyl] -4-phenylpiperazine, obtained in Reference Example 14 in the form of crystals, was reacted with 1-naphthalenesulfonyl chloride according to the method of Example XXXIV and the product thus obtained was treated with methyl iodide according to the procedure described in Example XXXV, to obtain N-methyl-N- {1- [p- (α-naphthalenesulfonyloxy) benzyl] -2- (4-phenylpiperazinyl) ethyl} -α naphthalene sulfonamide in a colorless, amorphous form.

Example LVIII

The amorphous compound obtained in example LVII was subjected to an alkaline hydrolysis according to the method described in example XXXVI to obtain N- [1- (p-hydroxybenzyl) -2- (4-phenylpiperazinyl) ethyl] -N-methyl-a -naphthalene sulfonamide in a colorless, amorphous form.

Reference Example 15 1- {[2-Amino-3- (p-hydroxy) phenyl] propyl} -4-benzyloxycarbonylpiperazine.

55 1.41 g of 1- (N-tert-butoxycarbonyltyrosyl) -4- (benzyloxycarbonyl) piperazine prepared by the method described in Reference Example 11 was dissolved in 5.6 ml of absolute ethyl acetate and 11.25 ml was added dropwise to this solution 4N hydrochloric acid in ethyl acetate with stirring 17 194549 added while cooling with ice for two minutes and the reaction mixture was stirred for 2 hours at room temperature. After the reaction was terminated, the solvent was evaporated under reduced pressure and to the residue was added 20 ml of 5% sodium bicarbonate in the form of an aqueous solution. The mixture was extracted with 30 ml and then with 20 ml of a mixed chloroform / methanol (9: 1) solvent and the extract was washed with an aqueous saturated solution of sodium chloride, dried over magnesium sulfate and filtered.

The filtrate was evaporated under reduced pressure to obtain a colorless foam. The foam was applied to a silica gel column and eluted with chloroform / methanol (6: 1) to obtain about 700 mg of the title compound in a colorless, amorphous form.

10

Example LIX

N- {2- (4-Benzyloxycarbonylpiperazinyl) -1- [p- (5-isoquinolinesulfonyloxy) benzyl] ethyl} -5-isoquinolinesulfonamide.

In 18 ml of absolute tetrahydrofuran was dissolved 680 mg of amorphous compound obtained in Reference Example 15, and 1.27 g of 5-isoquinolinesulfonyl chloride.HCl was added to the solution and then 3.20 ml of triethylamine was added dropwise with stirring and while cooling with ice for one minute and the mixture was stirred at room temperature for 150 minutes. The reaction mixture was diluted with 50 ml of chloroform, washed with water and the wash was extracted with 20 ml of chloroform. The combined organic extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and filtered and the filtrate was evaporated under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (100: 1) to obtain 987 mg of the title compound in a pale yellow, amorphous form.

Example LX

N- {2- (4-Benzyloxycarbonylpiperazinyl) -1- [p- (5-isoquinolinesulfonyloxy) benzyl] ethyl} -N-methyl-5-isoquinolinesulfonamide.

In 8.5 ml of absolute dimethylformamide, 852 mg of amorphous compound obtained in Example LIX was dissolved and 59 mg of 60% sodium hydride was added to the solution with stirring and cooling with ice, and further 113 µl of methyl iodide was added and the reaction mixture was stirred for two hours while cooling with ice. After the addition of 30 ml of ice water, the reaction mixture was extracted with 30 ml, 20 ml and 20 ml of ethyl acetate, the combined extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate, filtered and evaporated under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (100: 1) to obtain 679 mg of the title compound in a pale yellow, amorphous form.

Example LXI

N- {2 - [(4-Benzenesulfonyl) piperazinyl] -1- [p- (5-isoquinolinesulfonyloxy) benzyl] ethyl} -N-methyl-5-40 isoquinolinesulfonamide.

To 480 mg of the amorphous compound obtained in Example LX, 3 ml of a solution of 30% hydrogen bromide in acetic acid was added and the mixture was stirred at room temperature for 80 minutes. Then 50 ml of ether was added to the mixture which was then stirred. The resulting precipitate was collected and washed with ether and dried under reduced pressure to obtain 567 mg of N- {1- [p- (5-isoquinolinesulfonyloxy) benzyl] -2-piperazinyl-ethyl} -N-methyl-5- isoquinolinesulfonamide. HBr in the form of colorless crystals.

550 mg of crystals thus obtained were suspended in 10 ml of absolute tetrahydrofuran and, after stirring while cooling with ice, 84 µl of benzenesulfonyl chloride and 767 μΙ of triethylamine were added to the suspension and the reaction mixture was stirred at room temperature for 140 minutes. Then 50 ml of chloroform and 20 ml of water were added to the reaction mixture and after separation of the layers, the aqueous layer was extracted with 20 ml of chloroform. The chloroform layers were combined, washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and filtered. The filtrate was evaporated under reduced pressure and the resulting residue was applied to a silica gel column and eluted with chloroform / methanol (100: 1) to obtain 331 mg of the compound mentioned in the preamble in a pale yellow, amorphous form.

194549 18

Example LXII

N- {2 - [(4-Benzenesulfonyl) piperazinyl) -1- (p-hydroxybenzyl) ethyl) -N-methyl-5-isoquinolinesulfonamide.

In a mixed solution of 2.69 ml of methanol and 0.66 ml of tetrahydrofuran, 221 mg of amorphous compound obtained in Example LXI was dissolved and 0.33 ml of 2N sodium hydroxide as an aqueous solution was added to the solution. The reaction mixture was refluxed for 3.5 hours and to the mixture was added 30 ml of chloroform and 20 ml of 10% aqueous solution of ammonium chloride and the resulting layers were separated. The aqueous layer was extracted with 20 ml of chloroform and the chloroform layers were combined, washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and filtered. The filtrate was evaporated under reduced pressure and the resulting residue was applied to a silica gel column and eluted with chloroform / methanol (100: 1) to obtain 146 mg of the title compound in a colorless, amorphous form.

Reference Example 16 O-Benzyl-N-benzyloxycarbonyl tyrosinol.

27.25 g of O-benzyl-N-benzyloxycarbonyltyrosine methyl ester were dissolved in a mixed solvent of 185 ml of ethanol and 122 ml of tetrahydrofuran and 5.8 g of lithium chloride and 5.2 g of sodium borohydride were added to the solution while cooling with ice. The reaction mixture was stirred at room temperature for 18 hours and after the addition of 500 ml of saturated sodium chloride aqueous solution was extracted twice with 300 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure to obtain 25.4 g of the title compound in the form of colorless crystals.

Reference Example 17 1- [2-Benzyloxycarbonylamino-3- (p-benzyloxyphenyl) propyl] -4- (tert-butoxycarbonyl) piperazine.

In 70 ml of carbon tetrachloride, 5.6 g of crystals obtained in Reference Example 16 were dissolved, and after the addition of 4.5 g of triphenylphosphine, the mixture was refluxed for 20 hours. The reaction mixture was filtered to remove the insoluble material and the filtrate was evaporated under reduced pressure and the resulting residue was applied to a silica gel column and eluted with hexane / ethyl acetate (6: 1) to obtain 4.96 g of 2- benzyloxycarbonylamino-3- (p-benzyloxyphenyl) propyl chloride in the form of colorless crystals.

4.1 g of the above crystals and 2.23 g of N- (tert-butoxycarbonyl) piperazine were dissolved in 40 ml of dimethylformamide and 1.8 g of methyl iodide and 1.66 g of potassium carbonate were added to the solution and the mixture was stirred at 120 ° C for 3 hours. After the addition of 100 ml of saturated aqueous sodium chloride solution, the reaction mixture was extracted twice with 60 ml of chloroform and the extract was dried over magnesium sulfate and evaporated under reduced pressure. The resulting residue was applied to a silica gel column and eluted with hexane / ethyl acetate (2: 1) to obtain 2.69 g of the title compound in a colorless, amorphous form.

Example LXIII

N- {2- [4- (Tert-butoxycarbonyl) piperazinyl] -1- [p- (5-isoquinolinesulfonyloxy) benzyl] ethyl} -5-isoquinolinesulfonamide.

1.6 g of amorphous compound obtained in Reference Example 17 were dissolved in 25 ml of methanol and 1.0 g of 5% palladium on carbon was added to the solution. The mixture was stirred in a hydrogen 45 atmosphere for 20 hours and filtered to remove the insoluble material. The filtrate was evaporated under reduced pressure, the resulting residue dissolved in 30 ml of tetrahydrofuran and 2.8 g of 5-isoquinolinesulfonyl chloride.HCl and 4 ml of triethylamine were added to the solution while cooling with ice. After stirring at room temperature for 3 hours, 100 ml of water was added to the reaction mixture and the mixture was extracted twice with 70 ml of chloroform and the extract was dried over magnesium sulfate and evaporated under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (100: 1 to 50: 1) to obtain 1.27 g of the title compound in a yellow, amorphous form.

Example LXIV

55 In a mixed solvent of 7.5 ml of tetrahydrofuran and 7.5 ml of dimethylformamide, 940 mg of amorphous compound obtained in Example LXIII was dissolved and 67 mg of 60% sodium hydride and 0.11 ml of methyl iodide were successively added to the solution while cooling with ice and the mixture was stirred at room temperature at 19 194549 for one hour. After the addition of 30 ml of saturated aqueous solution of sodium chloride, the reaction mixture was extracted with 40 ml of ethyl acetate and the extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (60: 1) to give 723 mg of N- {2- [4- (tert-butoxycarbonyl) piperazinyl] - [p- (5-isoquinolinesulfonyloxy) ) benzyl] ethyl} -N-methyl-5-isoquinolinesulfonamide in a yellow, amorphous form.

Example LXV

To 720 mg of amorphous compound obtained in Example LXIV, 10 ml of 4N hydrochloric acid in ethyl acetate was added and the mixture was stirred at room temperature for one hour and evaporated under reduced pressure. To the mixture was added 30 ml of saturated aqueous solution of sodium bicarbonate and the reaction mixture was extracted twice with 20 ml of a mixed solvent of chloroform / isopropanol (5: 1), The extract was dried over magnesium sulfate and evaporated to dryness under reduced pressure to obtain 620 mg of N- {1- [p- (5-isoquinolinesulfonyloxy) benzyl] ethyl 2-piperazinyl} -N-methyl-5-isoquinolinesulfonamide in a yellow amorphous form.

Example LXVI

In 10 ml of methylene chloride was dissolved 620 mg of amorphous compound obtained in Example LXV, to the solution were added 0.29 ml of benzyl chloroformate and 0.4 ml of triethylamine while cooling with ice. The reaction mixture was stirred for 2 hours while cooling with ice and after the addition of 40 ml of a saturated aqueous solution of sodium chloride was extracted twice with 20 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure and the resulting residue was applied to a silica gel column and eluted with chloroform / methanol (60: 1) to give 660 mg of light yellow, amorphous product containing the same 1 H-NMR spectrum showed as the compound of example LX.

Example LXVII

In 6 ml of methanol was dissolved 650 mg of amorphous compound obtained in Example LXVI and to the solution was added 2 ml of 1N sodium hydroxide as an aqueous solution. The mixture was refluxed for 2 hours and after the addition of 30 ml of saturated aqueous sodium chloride solution was extracted twice with 200 ml of a mixed chloroform / isopropanol solvent (5: 1). The extract was dried over magnesium sulfate and evaporated under reduced pressure and the resulting residue was applied to a silica gel column and eluted with chloroform / methanol (80: 1 to 50: 1) to give 386 mg of N * [2- (4 -benzyloxycarbonylpiperazinyl) -1- (p-hydroxybenzyl) ethyl] -N-methyl-5-isoquinolinesulfonamide in a yellow, amorphous form.

Example LXVIII

In the amorphous compound obtained in Example LXIII, the protecting group of the piperazine 40 unit was removed according to the method described in Example LXV and the deprotected intermediate was treated according to the methods described in Examples LXVI and LXVII to obtain N- [ 2- (4-benzyloxycarbonylpiperazinyl) 1- (p-hydroxybenzyl) ethyl] -5-isoquinolinesulfonamide.

Example LXIX

The same procedure as described in Example LXVIII was repeated except that phenylpropionyl chloride was used instead of benzyl chloroformate to obtain N- {1- (p-hydroxybenzyl) -2- [4- (3-phenylpropyl) piperazinyl] ethyl} -5 isoquinolinesulfonamide.

Example LXX

The same procedure as described in Example LXVIII was repeated except that phenyl isocyanate was used instead of benzyl chloroformate to obtain N- [1- (p-hydroxybenzyl) -2- (4-phenylaminocarbonylpiperazinyl) ethyl] -5-isoquinolinesulfonamide in a yellow , amorphous form.

Example LXXI

The same procedure as described in Example LXVIII was repeated except that benzyl isocyanate was used instead of benzyl chloroformate to obtain N- [2- (4-benzylaminocarbonylpiperazinyl) -1- (p-hydroxybenzyl) ethyl] -5-isoquinolinesulfonamide in a yellow , amorphous form.

194549 20

Example LXXII

In 5 ml of chloroform, 200 mg of amorphous compound obtained in Example LXVII was dissolved, and to the solution was added 1 ml of methanol and also a solution of diazomethane in ether while cooling with ice. The reaction mixture was stirred for 90 minutes and then evaporated. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (80: 1 to 50: 1) to obtain 103 mg of N- [2- (4-benzyloxycarbonylpiperazinyl) -1- (p-methoxybenzyl) ethyl ] -N-methyl-5-isoquinolinesulfonamide.

Example LXXIII

The product of example LXVIII was treated according to the method described in example LXXII to obtain N- [2- (4-benzyloxycarbonylpiperazinyl) -1- (p-methoxybenzyl) ethyl] -5-isoquinolinesulfonamide.

Example LXXIV

N- {2- (4-Benzoylpiperazinyl) -1- [p- (5-isoquinolinesulfonyloxy) benzyl] ethyl} -N-methyl-5-isoquinolinesulfonamide. 764 mg of intermediate in the form of crystals obtained in Example LXI were suspended in 7 ml of tetrahydrofuran, 135 mg of benzoyl chloride was added to the suspension and 5 minutes later 1.1 ml of triethylamine were cooled with ice and the mixture was stirred for one hour. After adding chloroform and water, the reaction mixture was extracted three times with 30 ml of chloroform and the extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated to dryness under a reduced pressure. Of the resulting pale yellow residue, 0.82 g was applied to a silica gel column and eluted with chloroform / methanol (100: 1) to obtain 198 mg of the title compound in the colorless, amorphous form.

Example LXXV

N- [2- (4-Benzoylpiperazinyl) -1- (p-hydroxybenzyl) ethyl] -N-methyl-5-isoquinolinesulfonamide.

To 349 mg of the amorphous compound obtained in Example LXXIV, 4 ml of methanol, 1 ml of tetrahydrofuran and 0.5 ml of 2N sodium hydroxide were added and after refluxing for 2 hours, chloroform, water and sodium chloride were added to the reaction mixture. The reaction mixture was extracted three times with 30 ml of chloroform and the extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated under reduced pressure to obtain 0.25 g of yellow oil. The yellow oil was applied to a silica gel column and eluted with chloroform / methanol (50: 1) to obtain 145 mg of the title compound in the form of a colorless, amorphous form.

Example LXXVI

The same procedures as described in Examples LXXIV and LXXV were repeated sequentially, except that 470 mg of the crystalline intermediates in Example LXI and 113 mg of benzylsulfonyl chloride were used instead of benzoyl chloride to obtain 116 mg of N- [2- (4- benzylsulfonylpiperazinyl) -1 - (p-hydroxybenzyl) ethyl] -N-methyl-5-isoquinolinesulfonamide in a colorless, amorphous form.

Example LXXVII

The same procedures as described in Examples LXXIV and LXXV were repeated sequentially, except that 382 mg of the crystalline intermediates in Example LXI and 133 mg of 45-isoquinolinesulfonamide.HCl were used instead of benzoyl chloride to obtain N- {1- (p -hydroxybenzyl) -2- [4- (5-isoquinolinesulfonyl) piperazinyl] ethyl} -N-methyl-5-isoquinolinesulfonamide in a colorless, amorphous form.

Example LXXVIII

The same procedures as described in Examples LIX and LX were repeated sequentially, except that α-naphthalenesulfonyl chloride was used instead of 5-isoquinoline sulfonyl chloride.HCl to obtain N- {2- (4-benzyloxycarbonylpiperazinyl) -1- [p- ( α-naphthalenesulfonyloxy) benzyl] ethyl} -N-methyl-α-naphthalenesulfonamide in a colorless, amorphous form.

55 Example LXXIX

The amorphous compound obtained in Example LXXVIII was subjected to an alkaline hydrolysis according to the method described in Example LXII to obtain N- [2- (4-benzyloxycarbonyl-1945459)

piperazinyl) -1- (p-hydroxybenzyl) ethyl] -N-methyl-α-naphthalenesulfonamide in a colorless, amorphous form. Example LXXX

N- {1- [p- (5-isoquinolinesulfonyloxy) benzyl] -2- (4-phenyl homopiperazinyl) ethyl} -5-isoquinolinesulfonamide.

The same method as described in Example XXXIV was followed, except that 1- [2-amino-3- (p-hydroxyphenyl) propyl] -4-phenyl homopiperazine was used instead of 1- [2-amino-3- (p-) hydroxyphenyl) propyl 4-phenylpiperazine to obtain the title compound in a yellow amorphous form.

Example LXXXI

The amorphous compound obtained in Example LXXX was treated with methyl iodide according to the method described in Example XXXV to obtain N- {1- [p- (5-isoquinolinesulfonyloxy) benzyl] -2- (4-phenylhomopiperazinyl) ethyl} -N- methyl 5-isoquinino sulfonamide in a yellow, amorphous form.

The amorphous compounds obtained in Examples LXXX and LXXXI were subjected to alkaline hydrolysis according to the method described in Example XXXVI to obtain the following two compounds.

Example LXXXII

N- [1- (p-Hydroxybenzyl) * 2- (4-phenylhomopiperazinyl) ethyl] -5-isoquinolinesulfonamide.

Yellow crystals; 20

Example LXXXIII

N- [1- (p-Hydroxybenzyl) -2- (4-phenylhomopiperazinyl) ethyl] -N-methyl-5-isoquinolinesulfonamide.

Yellow amorphous substance.

Reference Example 18 N-tert-butoxycarbonyl-4-hydroxypiperidine.

7.14 g of 4-piperidone monohydrate hydrochloride was dissolved in 50 ml of dimethylformamide and 10 ml of water and 25 ml of diisopropylethylamine and 9.5 g of di-tert-butyldicarbonate were added to the solution at room temperature with stirring and the reaction mixture was added for 4 stirred for hours. After addition of water and saturation with sodium chloride, the reaction mixture was extracted twice with 300 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure to obtain 9.6 g of residue which was then dissolved in 100 ml of ethanol. To the solution was added 1.83 g of sodium borohydride with stirring and cooling with ice and the mixture was stirred for 90 minutes under the same conditions and then for 30 minutes at room temperature. After adding a saturated aqueous solution of sodium chloride, the reaction mixture was made alkaline with sodium bicarbonate and extracted twice with 600 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure and the resulting residue was subjected to a silica gel column treatment and eluted with hexane / ethyl acetate (2: 1) to obtain 8.03 g of the title compound. in a colorless, amorphous form.

40

Reference Example 19 4- (p-Methylbenzyloxy) piperidine.

2.0 g of amorphous compound obtained in Reference Example 18 were dissolved in 20 ml of dimethylformamide, and 0.48 g of 60% sodium hydride was added to the solution in an ice bath. After removal from the ice bath 45, the reaction mixture was stirred at room temperature for 30 minutes and after addition of 1.54 g of p-methylbenzyl chloride further stirred for 18 hours. The reaction mixture was poured onto ice, saturated with sodium chloride and extracted twice with 150 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure and the resulting residue was applied to a silica gel column and eluted with hexane / ethyl acetate (5: 1) to give 50 1.27 g of N-tert-butoxycarbonyl-4-p -methylbenzyloxypiperidine. This compound was dissolved in 3 ml of ethyl acetate and after addition of 12 ml of 3N hydrochloric acid in ethyl acetate at room temperature with further stirring for 17 hours, the solvent was evaporated under reduced pressure. The resulting residue was dissolved in water and the solution was made alkaline with sodium bicarbonate, saturated with sodium chloride, and extracted twice with 200 ml of chloroform. The extract was dried over 55 magnesium sulfate and the solvent was evaporated under reduced pressure to obtain 830 mg of the title compound in a colorless, amorphous form.

194549 22

Reference Example 20 N-Tert-butoxycarbonyl-o- (2-methoxyethoxymethyl) tyrosine methyl ester.

13.39 g of N-tert-butoxycarbonyl tyrosine methyl ester was dissolved in 65 ml of tetrahydrofuran and 65 ml of dimethylformamide and 1.9 g of 60% sodium hydride was added to the solution with stirring in an ice bath. After removal from the ice bath, the mixture was stirred at room temperature for 30 minutes and after adding 5.4 g of methoxyethoxymethyl chloride while cooling with ice, stirred for 15 hours while the mixture was warmed to room temperature. The reaction mixture was poured out on ice, saturated with sodium chloride and extracted twice with 800 ml of chloroform. The extract was dried over magnesium sulfate and the solvent was evaporated under reduced pressure and the resulting residue was applied to a silica gel column and eluted with hexane / ethyl acetate (4: 1) to obtain 13.85 g of the title mentioned in the preamble. compound in a colorless, amorphous form.

Reference Example 21 2- (N-Tert-butoxycarbonylamino) -1-chloro-3- [p- (2-methoxyethoxymethoxy) phenyl] propane.

13.85 g of amorphous compound obtained in Reference Example 20 was dissolved in 90 ml of ethanol and 60 ml of tetrahydrofuran and 3.11 g of lithium chloride and 2.77 g of sodium borohydride were added to the solution with stirring in an ice bath. After removing the ice bath, the mixture was stirred at room temperature for 16 hours and after adding the saturated aqueous solution of sodium chloride, the reaction mixture was made alkaline with sodium bicarbonate and extracted twice with 800 ml of chloroform. The extract was dried over magnesium sulfate and the solvent was evaporated under reduced pressure to give 11.73 g of N-tert-butoxycarbonyl-o- (2-methoxyethoxymethyl) tyrosinol. This compound was dissolved in 120 ml of carbon tetrachloride and 10 g of triphenylphosphine was added to the solution. The mixture was refluxed for 3 hours and further heated at 80 ° C for 17 hours. The solvent was evaporated under reduced pressure and the resulting residue was applied to a silica gel column and eluted with chloroform / methanol (100: 1), followed by eluting with hexane / ethyl acetate (4: 1) to obtain 7.22 g of the compound mentioned in the preamble in a colorless, amorphous form.

Reference Example 22 N- {2- (Tert-butoxycarbonylamino) -3- [p- (2-methoxyethoxymethoxy) phenyl] propyl} -4- (p-methylbenzyloxy-piperidine.

In 25 ml of dimethylformamide, 1.56 g of amorphous compound obtained in Reference Example 21 was dissolved, to the solution was added 0.83 g of amorphous compound obtained in Reference Example 19, 0.67 g of potassium carbonate and 0.67 g of sodium iodide and the mixture was stirred at 100 ° C for 2 hours and after addition of the saturated aqueous sodium chloride solution extracted twice with 150 ml of chloroform. The extract was dried over magnesium sulfate and the solvent was evaporated under reduced pressure. The resulting residue was applied to a column of silica gel and eluted with chloroform / methanol (100: 1) to obtain 490 mg of the title compound in a colorless, amorphous form.

Example LXXXIV

N- {1- [p- (5-isoquinolinesulfonyloxy) benzyl] -2- (4- (p-methylbenzyloxy) piperidino] ethyl} -5-isoquinolinesulfonamide.

45 In 1 ml of ethyl acetate was dissolved 490 mg of amorphous compound obtained in Reference Example 22 and to the solution was added 5 ml of 3N hydrochloric acid in ethyl acetate at room temperature with stirring and the reaction mixture was stirred for one hour. After evaporation of the solvent, the resulting residue was made alkaline with sodium bicarbonate as an aqueous solution and the mixture was saturated with sodium chloride, washed with a small amount of methanol and extracted twice with 100 ml of 50 chloroform. The extract was dried over magnesium sulfate and the solvent was evaporated to give a residue consisting of N- [2-amino-3- (p-hydroxyphenyl)] propyl-4- (p-methylbenzyloxy) piperidine. The residue was dissolved in 7 ml of tetrahydrofuran and to the solution were added 545 mg of 5-isoquinolinesulfonyl chloride.HCl and 450 mg of triethylamine at room temperature with stirring. The reaction mixture was stirred for 18 hours, made alkaline with an aqueous solution of 55 sodium bicarbonate and extracted twice with 100 ml of chloroform. The extract was dried over magnesium sulfate and the solvent was evaporated under reduced pressure and the resulting residue was applied to a silica gel column and eluted with chloroform / methanol (30: 1) to give 23 194549 572 mg of the title compound. in a colorless, amorphous form.

Example LXXXV

N- {1- (p-Hydroxybenzyl) -2- [4- (p-methylbenzyloxy) piperidino] ethyl} isoquinolinesulfonamide.

In 2.5 ml of methanol and 2.5 ml of tetrahydrofuran, 400 g of amorphous compound obtained in Example LXXXIV were dissolved, 4 ml of 1N sodium hydroxide solution was added to the solution and the mixture was heated under reflux for 2 hours. After cooling, the reaction mixture was diluted with water, acidified with citric acid, and then made alkaline with sodium bicarbonate and extracted twice with 100 ml of chloroform. The extract was dried over magnesium sulfate and the solvent was evaporated under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (20: 1) to obtain 172 mg of the title compound in a colorless, amorphous form.

Example LXXXVI

The same procedure as described in Example LXXXIV was repeated, except that N- {2- (tert-butoxycarbonylamino) -3- [p- (2-methoxyethoxymethoxy) phenyl] propyl} -4 * (3,4-dichlorobenzyloxy) piperidine used instead of N- {2- (tert-butoxycarbonylamino) -3- [p- (2-methoxyethoxymethoxy) phenyl] propyl} -4- (p-methylbenzyloxy) piperidine to give N- {1- [p-5 - isoquinolinesulfonyloxy) benzyl] -2- [4- (3,4-dichlorobenzyloxy) piperidino] ethyl} -5-isoquinolinesulfonamide in a colorless, amorphous form.

20

Example LXXXVII

The amorphous compound obtained in Example LXXXVI was subjected to an alkaline hydrolysis according to the method in Example LXXXV to give N- {1- (p-hydroxybenzyl) -2- [4- (3,4-dichlorobenzyloxy) piperidino] ethyl} - 5-isoquinolinesulfonamide in a colorless, amorphous form.

25

Reference example 23

The same procedure as described in Reference Example 21 was repeated except that N-benzyloxycarbonyl-o-benzyltyrosine methyl ester was used instead of N-tert-butoxycarbonyl-o- (2-methoxyethoxymethyl) tyrosine methyl star to obtain 2-benzyloxycarbonylamino-3- (p -30 benzyloxyphenyl) -1-chloropropane in a colorless, amorphous form.

To the amorphous compound thus obtained, N-phenylpiperazine was added and the same procedure as described in Reference Example 21 was repeated to give 1- {2- (benzyloxycarbonylamino) -3- (p-benzyloxyphenyl) propyl) -4-phenylpiperazine in a colorless , amorphous form.

Example LXXXVIII

N- {1- [p-Benzytoxy) benzyl] -2- (4-phenylpiperazinyl) ethyl} -5-isoquinolinesulfonamide.

500 mg of 1-substituted 4-phenylpiperazine obtained in Reference Example 23 were dissolved in 1 ml of acetic acid and 2 ml of 30% hydrobromic acid in acetic acid was added to the solution and the mixture was stirred for 10 minutes and poured onto ice. After the saturated aqueous solution of sodium thiosulfate was added, the reaction mixture was made alkaline with a saturated aqueous solution of sodium bicarbonate and extracted twice with 150 ml of chloroform. The extract was dried over magnesium sulfate and the solvent was evaporated under reduced pressure and the resulting residue was applied to a silica gel column and eluted with chloroform / methanol (30: 1), to obtain 235 mg of 1- {2-amino-3 - (p-benzyloxyphenyl) propyl} -4-phenylpiperazine. 235 mg of this compound were dissolved in 5 ml of tetrahydrofuran and 195 mg of 5-isoquinolinesulfonyl chloride.HCl and 178 mg of triethylamine were added to the solution at room temperature with stirring and the mixture was stirred for 16 hours. After addition of the saturated aqueous NaCl solution, the reaction mixture was made alkaline with an aqueous solution of sodium bicarbonate and extracted twice with 150 ml of chloroform. The extract was dried over magnesium sulfate and the solvent was evaporated under reduced pressure and the resulting residue was applied to a silica gel column and eluted with chloroform / methanol (50: 1) to obtain 280 mg of N- {1 - [(4 -benzyloxy) benzyl] -2- (4-phenylpiperazinyl) ethyl} -5-isocyanine sulfonamide in a colorless, amorphous form.

Example LXXXIX

55 N- {2- [4- (3,4-Dichlorobenzyloxy) piperidino] -1- [p- (5-isoquinolinesuffonyloxy) benzyl] ethyl} -N-methyl-5-isoquinolinesulfone amide.

1.70 g of amorphous compound obtained in Example LXXXVI was dissolved in 10 ml of dimethyiformamide and 194549 24 mg of sodium hydride was added to the solution with stirring in an ice bath and after stirring. .

the ice bath was removed for 10 minutes. The reaction mixture was stirred at room temperature for 4 hours and after adding 150 ml of ethyl acetate washed three times with water and the washed material was extracted with 50 ml of ethyl acetate. The extracts were combined and washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and the solvent was evaporated under reduced pressure. The resulting residue was applied to a column of silica gel (silica gel: Fuji Debison Kagaku, BW-820MH) and eluted with 1% methanol in chloroform to obtain 1.30 g of the title compound in a colorless, amorphous form.

10

Example XC

N- {2- [4- (3,4-Dichlorobenzyloxy) piperidino] -1- (p-hydroxybenzyl) ethyl} -N-methyl-5-isoquinolinesulfonamide.

1.04 g of amorphous compound obtained in Example LXXXIX were dissolved in 10 ml of dimethyl sulfoxide, a solution of 152 mg of sodium hydroxide in 2 ml of water was added to the solution and the mixture was stirred at 80 ° C for 2 hours. After addition of 150 ml of ethyl acetate, the reaction mixture was washed twice with 100 ml of water and the wash was extracted with 50 ml of ethyl acetate. The ethyl acetate layers were combined, washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting residue was applied to a silica gel column and eluted with 1% methanol in chloroform to give 650 mg of the title compound in a colorless, amorphous form.

Example XCI

N- {2- [4- (3,4-Dichlorobenzyloxy) piperidine] -1- (p-methoxybenzyl) ethyl} -N-methyl-5-isoquinolinesulfonamide.

350 mg of amorphous compound obtained in Example XC were dissolved in 10 ml of dimethylformamide and 82.8 mg of methyl iodide was added to the solution with stirring in an ice bath. The mixture was stirred for 30 minutes and, after removing the ice bath, further stirred for 2 hours at room temperature. 100 ml of ethyl acetate was added to the reaction mixture and the mixture was washed three times with 50 ml of water. The wash was extracted with 50 ml of ethyl acetate, and the layer of ethyl acetate was washed with water. The ethyl acetate layers were combined, washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting residue was applied to a column of silica gel and eluted with 1% methanol in chloroform to obtain 297 mg of the title compound in a colorless, amorphous form.

35

Example XCU

The same procedures as described in Examples XXXIV and XXXV were repeated sequentially, except that 1- [2-amino-3- (p-hydroxyphenyl) propyl] -4-phenylpiperidine was used instead of 1- [2-amino-3- ( p-hydroxyphenyl) propyl] -4-phenylpiperazine to give N- {1 - [p- (5-isoquinolinesulfonyloxy) benzyl] -2- (4-40 phenylpiperidino) ethyl} -N-methyl-5-isoquinolinesulfonamide in a colorless , amorphous form.

Example XClII

The amorphous compound obtained in Example XCII was subjected to an alkaline hydrolysis according to the method of Example XXXVI to obtain N- [1- (p-hydroxybenzyl) -2- (4-phenylpiperidino) ethyl] -N-45 methyl-5- isoquinolinesulfonamide in a colorless, amorphous form.

Example XCIV

The same procedures as described in Examples XXXIV and XXXV were repeated sequentially, except that 1- [2-amino-3- (phydroxyphenyl) propyl] -4,4-ethylenedioxypiperidine was used instead of 50 1- [2-amino-3 - (p-hydroxyphenyl) propyl] -4-phenylpiperazine to give N- {2- (4,4-ethylenedioxypiperidino) -1- [p- (5-isoquinolinesulfonyloxy) benzyl] ethyl} -N-methyl-5-isoquinolinesulfonamide in a colorless, amorphous form.

Example XCV

55 N- {1- [p- (5-isoquinolinesulfonyloxy) benzyl] -2- (4-oxopiperidino) ethyl} -N-methyl-5-isoquinolinesulfonamide.

In 50 ml of 3N hydrochloric acid, 2.57 g of the product from Example XCIV was dissolved and after refluxing for 6 hours and then cooling, the reaction mixture was made alkaline with a saturated, aqueous solution of sodium bicarbonate and extracted twice with 200 ml. chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure to remove the solvent and the resulting residue was applied to a silica gel column and eluted with chloroform / methanol (50: 1) to obtain 2.22 g of the title. said compound in a colorless, amorphous form.

Example XCVI

N- {2- [4- (N'-Benzyl-N'-methylamino) piperidino] -1- [p- (5-isoquinolinesulfonyloxy) benzyl] ethyl} -N-methyl-5-isoquinolinesulfonamide.

1.0 g of the product from Example XCV was dissolved in 15 ml of methanol, 270 mg of benzylmethylamine and 120 mg of sodium cyanoborium hydride were added to the solution with stirring and the reaction mixture was stirred for 18 hours. After addition of a saturated aqueous solution of sodium bicarbonate, the reaction mixture was extracted twice with 150 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure to remove the solvent and the resulting residue was applied to a silica gel column and eluted with chloroform / methanol (30: 1) to obtain 380 mg of the title mentioned in the preamble. compound in a colorless, amorphous form.

Example XCVII

N- {2- [4- (N-Benzyl-N-methylamino) piperidino] -1- (p-hydroxybenzyl) ethyl} -N-methyl-5-isoquinolinesulfonamide. In 2 ml of tetrahydrofuran and 2 ml of methanol was dissolved 380 mg of amorphous compound obtained in Example XCVI, to the solution was added 4 ml of 1N sodium hydroxide as an aqueous solution and the reaction mixture was heated under reflux for 3 hours and cooled. The reaction mixture was poured into water, acidified with citric acid and then made alkaline with sodium bicarbonate, washed with a small amount of methanol and extracted twice with 100 ml of chloroform. The extract was dried over magnesium sulfate and evaporated to remove the solvent under a reduced pressure and the resulting residue was applied to a silica gel column and eluted with chloroform / methanol (20: 1) to obtain 147 mg of the title mentioned in the preamble. compound in a colorless, amorphous form.

30

Example XCVIII

The same operations as described in Examples XCVI and XCVII were repeated sequentially, except that benzylamine was used instead of benzylmethylamine to give N- {2- [4- (N-benzylamino) piperidino] -1- (p-hydroxybenzyl) ethyl } -N-methyl-5-isoquinolinesulfonamide in a colorless, amorphous form.

Example IC

N- {2- (4-Hydroxypiperidino) -1- [p- (5-isoquinolinesulfonyloxy) benzyl] ethyl} -N-methyl-5-isoquinolinesulfonamide. 200 mg of amorphous compound obtained in Example XCV were dissolved in 5 ml of methanol, 35.2 mg of sodium borohydride was added in small amounts at room temperature with stirring and the mixture was stirred and evaporated for 2 hours to remove the solvent. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (100: 4) to obtain 116 mg of the title compound in the form of a light yellow oil.

45

Example C.

N- {2- (4-Hydroxypiperidino) -1- [p- (5-isoquinolinesulfonyloxy) benzyl] ethyl} -N-methyl-5-isoquinolinesulfonamide.

150 mg of the oil obtained in Example 1C were dissolved in 3 ml of methanol, 1 ml of 10% sodium hydroxide as aqueous solution was added to the solution, and the reaction mixture was refluxed for 2 hours and evaporated to remove the solvent under a reduced pressure, the resulting residue was acidified with citric acid and then made alkaline with an aqueous solution of sodium bicarbonate and the mixture was extracted three times with 20 ml of chloroform. The extract was dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure and the resulting residue was subjected to silica gel preparative thin layer 55 chromatography and separated by chloroform / methanol (20: 1) to obtain 87 mg of the in the preamble in a colorless, amorphous form.

194549 26

Example C1 N- [1- (p-Hydroxybenzyl) -2- (4-hydroxy piperidino) ethyl] -N-m ethyl 5-isoquinolinesulfonamide.

In 5 ml of a mixture of ethyl acetate / methanol (1: 1), 100 mg of amorphous compound obtained in Example C was dissolved, an excess amount of diazomethane in ether was added to the solution and the reaction mixture was stored overnight at room temperature. The solvent in the reaction mixture was evaporated under reduced pressure and the resulting residue was subjected to silica gel preparative thin layer chromatography and separated by chloroform / methanol (20: 1) to obtain 80.4 mg of the title compound in a colorless, amorphous form.

Example CU

N- [1- (p-Acetoxybenzyl) -2- (4-acetoxypiperidino) ethyl] -N-methyl-5-isoquinolinesulfonamide.

230 mg of the amorphous compound obtained in Example C were dissolved in 2 ml of pyridine, 1 ml of acetic anhydride was added to the solution and the reaction mixture was stored overnight at room temperature. To the mixture was added 20 ml of ice water and the mixture was stirred for one hour and extracted twice with 20 ml of ethyl acetate. The extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated to remove the solvent under a reduced pressure. The resulting residue was applied to a column of silica gel and eluted with chloroform / methanol (100: 1) to obtain 234.3 mg of the title compound in a colorless, amorphous form.

20

Example Clll

The same procedure as described in Example LXXXIV was repeated except that N- {2- (tert-butoxycarbonylamino) -3- [p- (2-methoxyethoxymethoxy) phenyl] propyl} -4-acetoxypiperidine prepared in a similar manner was used instead of N- {2- (tert-butoxycarbonylamino-3- [p- (2-25 methoxyethoxymethoxy) phenyl] propyl} -4- (p-methylbenzyloxy) piperidine to give N- {2- [4-acetoxypiperidino ] -1 - [p- (5-isoquinolinesulfonyloxy) benzyl] ethyl} -5-isoquinolinesulfonamide in a colorless, amorphous form.

Example CIV

N- {2- [4-Hydroxypiperidino] -1- [p- (5-isoquinolinesulfonyloxy) benzyl] ethyl} -5-isoquinolinesulfonamide.

1.5 g of the product obtained in Example C11 were dissolved in 8 ml of methanol, 8 ml of 1N aqueous solution of sodium hydroxide was added to the solution, and the mixture was stirred for 2 hours and, after adding water, extracted twice with 100 ml of chloroform . The extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated to remove the solvent under a reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (30: 1) to obtain 800 mg of the title compound in a colorless, amorphous form.

Example CV

40 N-3,4-Dichlorobenzyl-N- [1- (p-hydroxybenzyl) -2- (4-hydroxy piperidino) ethyl] -5-isoquinolinesulfonamide.

400 mg of amorphous compound obtained in Example CIV were dissolved in 5 ml of dimethylformamide, 130 mg of 3,4-dichlorobenzyl chloride and 28 mg of 60% sodium hydride were added to the solution with stirring and cooling with ice, and the mixture was warmed to room temperature and stirred for 18 hours. After adding a saturated aqueous solution of sodium chloride, the reaction mixture was extracted twice with 100 ml of chloroform and the extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure pressure. The resulting residue was applied to a column of silica gel and eluted with chloroform / methanol (30: 1) to obtain 228 mg of N- (3,4-dichlorobenzyl) -N- {2- (4-hydroxypiperidino) -1- [ p- (5-isoquinoline sulfonyloxy) benzyl] ethyl} -5-isoquinoline sulfonamide.

50 In 1.5 ml of methanol was dissolved 228 mg of the above-mentioned compound, to the solution was added 1 ml of 1N sodium hydroxide as an aqueous solution and the mixture was refluxed for 3 hours and then cooled and, after dilution with water, acidified with citric acid and then made alkaline with sodium bicarbonate and extracted twice with 100 ml of chloroform.

The extract was dried over magnesium sulfate and evaporated to remove the solvent and the resulting residue was applied to a silica gel column and eluted with chloroform / methanol (20: 1) to obtain 162 mg of the title compound in a colorless, amorphous form.

27 194549

Example CVI

N- [1- (p-Hydroxybenzyl) -2- (4-hydroxy piperidino) ethyl] -N-p methylbenzyl-5-isoquinolinesulfonamide.

The same process as described in Example CV was repeated, except that 4-methylbenzyl chloride was used instead of 3,4-dichlorobenzyl chloride to obtain the title compound in a colorless, amorphous form.

Example CVII

N- {2- (4-Hydroxypiperidino) -1- [p- (5-isoquinolinesulfonyloxy) benzyl] ethyl} -N-methyl-5-isoquinolinesulfonamide. The amorphous compound obtained in Example C11 was treated with methyl iodide according to the method described in Example LXXXIX and the intermediate was subjected to alkaline hydrolysis according to the method described in Example CIV to obtain the same product as indicated in Example IC.

Reference Example 24 1 - (N-Benzyloxycarbonyltyrosyl) -4-phenylpiperazine.

12.3 g of N-benzyloxycarbonyltyrosine and 6.6 g of N-phenylpiperazine were dissolved in 150 ml of methylene chloride and 8.4 g of DCC were added to the solution and the mixture was stirred at room temperature for 5 hours. The precipitated insoluble material was filtered off and the filtrate was evaporated under reduced pressure and the resulting residue was applied to a column of silica gel eluted with hexane / ethyl acetate (1: 1 to 1: 2) to obtain 10.5 g of the in the preamble in a colorless, amorphous form.

Example CVIII

1- [N, 0-Bis (5-isoquinolinesulfonyl) tyrosyl] -4-phenylpiperazine.

In 50 ml of methanol was dissolved 4.59 g of amorphous compound obtained in Reference Example 24, and to the solution was added 3 g of 5% palladium on carbon and the mixture was stirred for 17 hours at room temperature in a hydrogen atmosphere. The resulting insoluble material was filtered off and the filtrate evaporated under reduced pressure to obtain a residue which was then suspended in 50 ml of chloroform. To the suspension was sequentially added 5.7 g of 5-isoquinoline sulfonyl chloride / HCl and 10 ml of triethylamine while cooling with ice, and then the mixture was stirred at room temperature for 3 hours. After adding 200 ml of water, the mixture was extracted twice with 100 ml of chloroform, the extract dried over magnesium sulfate and evaporated under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (80: 1 to 30: 1) to obtain 5.46 g of the title compound in a yellow, amorphous form.

Example CIX

1- [N, O-Bis (5-isoquinolinesulfonyl) -N-methyl tyrosyl] -4-phenylpiperazine.

In 30 ml of dimethylformamide was dissolved 2.27 g of amorphous compound obtained in Example CVIII, to 40 the solution was added successively 160 mg of 60% sodium hydride and 0.3 ml of methyl iodide while cooling with ice and the mixture was stirred for 90 minutes while cooling with ice. After addition of 80 ml of water, the reaction product was extracted with 100 ml of ethyl acetate and the extract was washed with 80 ml of saturated aqueous sodium chloride solution, dried over magnesium sulfate and evaporated under reduced pressure. The resulting residue was applied to a column of 45 silica gel and eluted with chloroform / methanol (60: 1) to obtain 1.8 g of the title compound in a yellow, amorphous form.

Example CX

1- [N- (5-Isoquinolinesulfonyl) -N-methyl-tyrosyl] -4-phenyl-piperazine.

50 1.15 g of amorphous compound obtained in Example CIX was suspended in 20 ml of methanol, 2 ml of 2N sodium hydroxide dissolved in water was added to the suspension and the mixture was boiled for 90 minutes. After addition of 100 ml of water, the reaction mixture was extracted twice with 50 ml of chloroform and the extract was dried over magnesium sulfate and evaporated under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / 55 methanol (80: 1 to 50: 1) to obtain 820 g of the title compound in a colorless, amorphous form.

194549 28

Example CXI

The product of Example XVIII was subjected to alkaline hydrolysis according to the method described in Example CX to obtain 1- [N- (5-isoquinolinesulfonyl) tyrosyl] -4-phenylpiperazine in a yellow, amorphous form.

5

Reference Example 25 N-Benzyloxycarbonyl Homopiperazine.

To 230 ml of dimethylformamide were added 25 g of homopiperazine and 5.4 g of sodium bicarbonate and then 25 ml of water, followed by the dropwise addition of 10 g of benzyloxycarbonyl chloride with stirring and cooling with ice and the mixture was stirred at room temperature overnight. After evaporation of dimethylformamide under reduced pressure, the reaction mixture was extracted three times with 100 ml of chloroform and the extract was dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (9: 1) to obtain 9 g of the title compound in the form of a light yellow liquid.

Reference Example 26 1- [N- (Tert-butoxycarbonyl) -N-methyl] tyrosyl-4-benzyloxycarbonyl homopiperazine.

In 70 ml of methylene chloride, 1 g of N-tert-butoxycarbonyl-N-methyl tyrosine was dissolved and after adding 793 mg of N-benzyloxycarbonyl homopiperazine and adding 837 mg of DCC at room temperature with stirring, the mixture was stirred at room temperature for a night. The solvent was evaporated under reduced pressure and benzene was added to the resulting residue. Insoluble material was filtered off and the filtrate was applied to a column of silica gel and eluted with hexane / ethyl acetate (6: 4 to 6: 5) to obtain 1.06 g of the title compound in the form of a pale yellow oil.

Reference Example 27 1- {3 '- (p-Acetoxyphenyl) -2' [N - (tert-butoxycarbonyl) -N-methylamino] propyl} -4-benzyloxycarbonyl homopiperazine.

In 60 ml of absolute tetrahydrofuran, 3.56 g of the product obtained in Reference Example 26 was dissolved, 20 ml of 1.0 M borane in tetrahydrofuran was added with stirring and cooling with ice and the mixture was stirred overnight at room temperature. . The solvent was evaporated under reduced pressure and the resulting residue was dissolved in 10 ml of pyridine. To the solution was added 5 ml of acetic anhydride and the mixture was stored overnight at room temperature. After adding ice, the mixture was stirred for 30 minutes and extracted twice with 60 ml of chloroform. The extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting residue was applied to a column of silica gel and eluted with chloroform / methanol (100: 1), to obtain 2.0 g of the title compound in a pale yellow, amorphous form.

Example CXII

N- [1- (p-Acetoxybenzyl) -2- (4-benzyloxycarbonylhomopiperazinyl) ethyl] -N-methyl-5-isoquinolinesulfonamide.

1 g of amorphous compound obtained in Reference Example 27 was dissolved in 28 ml of methylene chloride, 45 ml of 2,6-lutidine and then 2 ml of tert-butyldimethylsilyl trifluoromethanesulfonate were added with stirring at room temperature and the reaction mixture was stirred for 16 hours. After adding ice, the reaction mixture was extracted twice with 70 ml of ethyl acetate and the extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. To the obtained residue 50, 20 ml of tetrahydrofuran and 4.28 ml of 1.0 M tetrabutylammonium fluoride in tetrahydrofuran were added with stirring and the reaction mixture was stirred at room temperature for 40 minutes. After adding ice, the reaction mixture was extracted twice with 70 ml of chloroform and the extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (95: 5 to 90:10) to give 723 mg of 1- [3 '- (p-acetoxyphenyl) -2' - (N-methylamino) propyl] -4-benzyloxycarbonyl homopiperazine.

29 194549

723 mg of the above compound was dissolved in 25 ml of dimethylformamide and 401 mg of triethylamine and then 564 mg of 5-isoquinolinesulfonyl chloride.HCl were added to the mixture with stirring and cooling with ice and the mixture was stirred overnight at room temperature. After addition of water, the reaction mixture was extracted twice with 70 ml of ethyl acetate and the extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (100: 1) to obtain 796 mg of the title compound in a pale yellow, amorphous form.

Example CXIII

N- [2- (4-Benzyloxycarbonylhomopiperazinyl) -1- (p-hydroxybenzyl) ethyl] -N-methyl-5-isoquinolinesulfonamide.

400 mg of amorphous compound obtained in Example CXII were dissolved in 10 ml of methanol, 2 ml of 10% sodium hydroxide was added to the solution and the mixture was stirred for 10 minutes. The reaction mixture was acidified with an aqueous solution of citric acid and then made alkaline with a saturated aqueous solution of sodium bicarbonate and extracted twice with 50 ml of chloroform. The extract was dried over magnesium sulfate and evaporated to remove the solvent under a reduced pressure. The resulting residue was applied to a column of silica gel and eluted with chloroform / methanol (100: 2) to obtain 339 mg of the title compound in a colorless, amorphous form.

20

Example CXIV

In 2 ml of acetic acid was dissolved 220 mg of amorphous compound obtained in Example CXIII, to the solution was added 6 ml of 25% hydrobromic acid in acetic acid and the mixture was stirred at room temperature for 20 minutes. 40 ml of dry ether was added to the reaction mixture to form a white precipitate, which was then made alkaline with a saturated aqueous solution of sodium bicarbonate and extracted twice with 20 ml of chloroform / isopropanol (5: 1). The extract was dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure and the resulting residue was applied to a silica gel column and eluted with chloroform / methanol (20:80 to 30:70) to give 67 mg of N- [ 1- (p-hydroxy) benzyl-2-homopiperazinylethyl] -N-methyl-5-iso-quinoline sulfonamide as a light yellow oil.

Reference Example 28 1-Benzyloxycarbonyl-4- (N-tert-butoxycarbonyltyrosyl) homopiperazine.

In 280 ml of tetrahydrofuran, 15.29 g of N-tert-butoxycarbonyl tyrosine and 12.73 g of N-benzyloxycarbonyl homopiperazine were dissolved, to the solution was added 8.09 g of 1-hydroxybenzotriazole hydrate and 11.77 g of DCC at room temperature with stirring and the reaction mixture was stirred for 16 hours. The reaction mixture was evaporated to remove the solvent under reduced pressure, benzene was added to the residue and the insoluble material was filtered off under reduced pressure and washed with benzene. The benzene layers were combined and evaporated under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (100: 1), to obtain 26.42 g of the title compound in a colorless, amorphous form.

Example CXV

45 N- {2- [4-Benzyloxycarbonyl) homopiperazinyl] -1- [p- (5-isoquinolinesulfonyloxy) benzyl] ethyl} -5-isoquinolinesulfonamide.

3.0 g of amorphous compound obtained in Reference Example 28 were dissolved in 20 ml of tetrahydrofuran, 24 ml of 1M borane in tetrahydrofuran was added to the solution and the mixture was stirred under a nitrogen atmosphere at room temperature for 15 hours. After the reaction was terminated, the solvent was evaporated under reduced pressure and to the resulting residue was added 3 g of potassium bicarbonate. The mixture was stirred at room temperature for 30 minutes and extracted twice with 200 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure to give 1-benzyloxycarbonyl-4- [2- (tert-butoxycarbonylamino) -3- (p-hydroxy-phenyl) propyl] homopiperazine.

This compound was dissolved in 6 ml of ethyl acetate, 30 ml of 4N hydrochloric acid in ethyl acetate was added to the solution and the mixture was stirred at room temperature for 30 minutes. The reaction mixture was evaporated under reduced pressure and the resulting residue was made alkaline with a saturated aqueous solution of sodium bicarbonate and extracted twice with 200 ml of chloroform. The extract was dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure to obtain 2.43 g of 1- [2-amino-3- (p-hydroxyphenyl) propyl] -4-benzyloxycarbonylhomopi perazine.

This intermediate was dissolved in 65 ml of tetrahydrofuran, to the solution was added 4.03 g of 5-isoquinolinesulfonyl chloride.HCl and 8.9 ml of triethylamine at room temperature with stirring and the mixture was stirred for 16 hours. After the addition of a saturated solution of sodium bicarbonate, the mixture was extracted twice with 300 ml of chloroform and the extract was dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (20: 1) to obtain 2.26 g of the title compound in a colorless, amorphous form.

Example CXVI

1.0 g of product obtained in Example CXV was dissolved in 5 ml of methanol and 5 ml of tetrahydrofuran, 10 ml of 1N sodium hydroxide was added to the solution and the mixture was heated under reflux for 2 hours and then cooled. The reaction mixture was acidified with citric acid and then made alkaline with sodium bicarbonate, the prepared insoluble material was then dissolved in methanol. The solution was extracted twice with 100 ml of chloroform and the extract was dried over magnesium sulfate and evaporated under reduced pressure. The resulting residue was applied to a column of silica gel and eluted with chloroform / methanol (20: 1) to give 458 mg of N- {2- (4-benzyloxycarbonylhomopiperazinyl) -1 - (p-hydroxybenzyl) ethyl} -5-isoquinolinesulfonamide in a colorless, amorphous form.

Example CXVII

N- {1- [p- (5-isoquinolinesulfonyloxy) benzyl] -2-homopiperazin-ethyl} -5-isoquinoline sulfonamide.

To 1.0 g of the product obtained in Example CXV, 6 ml of 30% hydrobromic acid in acetic acid was added at room temperature with stirring and the mixture was further stirred for 24 hours. After addition of 100 ml of ether, the reaction mixture was stirred for 30 minutes to form a salt, which was then collected by filtration, washed with ether and dried in a desiccator. The salt was dissolved in water and the solution was made alkaline with sodium bicarbonate and extracted twice with 100 ml of chloroform. The extract was dried with magnesium sulfate and evaporated to remove the solvent under reduced pressure, to obtain 830 mg of the title compound in a colorless, amorphous form.

Example CXVIII

N- {1- [p- (5-isoquinolinesulfonyloxy) benzyl] -2- [4- (3-phenylpropionyl) homopiperazinyl] ethyl} -5-isoquinolinesulfonamide.

420 mg of amorphous compound obtained in Example CXVII was dissolved in 6 ml of methylene chloride, 125 mg of 3-phenylpropionyl chloride and 100 mg of triethylamine were added to the solution at room temperature with stirring and the mixture was stirred for 17 hours. The reaction mixture was made alkaline with a saturated aqueous solution of sodium bicarbonate and extracted twice with 50 ml of chloroform. The extract was dried over magnesium sulfate and evaporated to remove the solvent. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (30: 1) to obtain 400 mg of the title compound in a 45 colorless, amorphous form.

Example CXIX

N- {1- (p-Hydroxybenzyl) -2- [4- (3-phenylpropionyl) homopiperazinyl] ethyl} -5-isoquinoline sulfonylamide.

400 mg of amorphous compound obtained in Example CXVIII 50 was dissolved in 2 ml of methanol and 2 ml of tetrahydrofuran, 4 mM N sodium hydroxide was added to the solution and the mixture was heated under reflux for 3 hours and then cooled. The reaction mixture was acidified with citric acid and then made alkaline with sodium bicarbonate to form an insoluble material which was then dissolved in a small amount of methanol and extracted twice with 50 ml of chloroform. The extract was dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting residue was applied to a column of silica gel and eluted with chloroform / methanol (30: 1), to obtain 230 mg of the title compound in a colorless, amorphous form.

31 194549

Reference Example 29 1-Benzyloxycarbonyl-4- (N-tert-butoxycarbonyl-o-methyl) tyrosyl homopiperazine.

In 25 ml of tetrahydrofuran and 25 ml of dimethylformamide, 5.0 g of amorphous compound obtained in Reference Example 28 was dissolved, to the solution was added 0.41 g of 60% sodium hydride with stirring and cooling with ice, and then the mixture was warmed to room temperature and for Stirred for 30 minutes. After adding 1.43 g of methyl iodide, the reaction mixture was stirred for 16 hours and extracted twice with 300 ml of chloroform after the addition of a saturated aqueous solution of sodium chloride. The extract was dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting residue was applied to a column of silica gel and eluted with chloroform / methanol (100: 1) to obtain 3.76 g of the title compound in a colorless, amorphous form.

Reference Example 30 1- (N-Tert-butoxycarbonyl-o-methyl) tyrosyl-4-phenylacetyl homopiperazine.

1.02 g of amorphous compound obtained in Reference Example 29 was dissolved in 25 ml of methanol, 250 mg of 5% palladium on carbon was added to the solution while cooling with ice, and after heating the mixture to room temperature, a catalytic reduction was carried out for 6 hours . The catalyst was filtered off and washed with methanol and the methanol solution was evaporated to give 800 mg of (N-tert-butoxycarbonyl-o-methyl) tyrosyl homopiperazine.

400 mg of the compound were dissolved in 6 ml of methylene chloride, 195 mg of phenylacetyl chloride and 190 mg of triethylamine were added to the solution and the mixture was stirred at room temperature for 24 hours. The reaction mixture was made alkaline with a saturated aqueous solution of sodium bicarbonate and extracted twice with 100 ml of chloroform and the extract was dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure.

The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (50: 1) to obtain 433 mg of the title compound in a colorless, amorphous form.

Example CXX

1- [N- (5-Isoquinolinesulfonyl) -o-methyl] tyrosyl-4-phenylacetyl homopiperazine.

In 1 ml of ethyl acetate, 433 mg of amorphous compound obtained in Reference Example 30 was dissolved, to the solution was added 4 ml of 4N hydrochloric acid in ethyl acetate, and after stirring for 30 minutes at room temperature, the solvent was evaporated under reduced pressure. A saturated aqueous solution of sodium bicarbonate was added to the resulting residue and the solution was extracted twice with 50 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure. To the resulting residue was added 6 ml of tetrahydrofuran as well as 275 mg of 5-isoquinolinesulfonyl chloride.HCl and 1.2 ml of triethylamine at room temperature with stirring and the mixture was further stirred for 18 hours. The reaction mixture was made alkaline with a saturated aqueous solution of sodium bicarbonate and extracted twice with 50 ml of chloroform and the extract was dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (30: 1) to obtain 439 mg of the title compound in a colorless, amorphous form.

Example CXXI

45 1- [N, O-Dimethyl-N- (5 * isoquinolinesulfonyl)] tyrosyl-4-phenylacetyl homopiperazine.

439 mg of amorphous compound obtained in Example CXX were dissolved in 2.5 ml of tetrahydrofuran and 2.5 ml of dimethylformamide, 30 mg of 60% sodium hydride was added to the solution while cooling with ice, and then the mixture was warmed to room temperature and stirred for 30 minutes . After the addition of 110 mg of methyl iodide, the reaction mixture was further stirred for 16 hours.

After adding a saturated aqueous solution of sodium chloride, the reaction mixture was extracted twice with 50 ml of chloroform and the extract was dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting residue was applied to a column of silica gel and eluted with chloroform / methanol (30: 1) to obtain 348 mg of the title compound in a colorless, amorphous form.

J

194549 32

Example CXXII

1-Benzyloxycarbonyl-4- [N, O-bis (5-isoquinolinesulfonyl) tyrosyl] homopiperazine.

In 6 ml of ethyl acetate was dissolved 6.44 g of amorphous compound obtained in Reference Example 28, to the solution was added 60 ml of 4N hydrochloric acid in ethyl acetate at room temperature with stirring and the mixture was stirred for 3 hours. The reaction mixture was evaporated under reduced pressure and, after adding benzene, evaporated again under reduced pressure to obtain 1-benzyloxycarbonyl-4-tyrosyl homopiperazine / hydrochloride in an amorphous form.

To this intermediate was added 130 ml of tetrahydrofuran as well as 7.88 g of 5-isoquinolinesulfonyl chloride.HCl and 18 ml of triethylamine and the mixture was stirred at room temperature for 18 hours. The reaction mixture was made alkaline with a saturated aqueous solution of sodium bicarbonate and extracted twice with 700 ml of chloroform and the extract was dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting residue was applied to a column of silica gel and eluted with chloroform / methanol (30: 1) to obtain 5.50 g of the title compound in a colorless, amorphous form.

15

Example CXXIII

1-Benzyloxycarbonyl-4- [N - (5-isoquinolinesulfonyl) tyrosyl] homopiperazine.

In 30 ml of methanol and 30 ml of tetrahydrofuran, 5.50 g of amorphous compound obtained in Example CXXII were dissolved, 60 ml of 1N sodium hydroxide was added to the solution and the mixture was heated under reflux for 2 hours and then cooled. The reaction mixture was acidified with citric acid and then made alkaline with sodium bicarbonate, the resulting insoluble material was dissolved with a small amount of methanol and the solution was extracted twice with 400 ml of chloroform. The extract was dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure and the resulting residue was applied to a silica gel column and eluted with chloroform / methanol (20: 1) to obtain 3.1 g of the the above compound in a colorless, amorphous form.

Reference Example 31 N-Benzyloxycarbonyl tyrosine and N-tert-butoxycarbonyl homopiperazine were treated according to the method described in Reference Example 28 to obtain 1- (N-benzyloxycarbonyl) tyrosyl-4-tert-butoxycarbonyl homopiperazine in a colorless, amorphous form.

Reference Example 32 1- (O-Acetyl-N-benzyloxycarbonyl) tyrosyl-4-tert-butoxycarbonyl homopiperazine.

In 7 ml of pyridine, 690 mg of amorphous compound obtained in Reference Example 31 was dissolved and 3.5 ml of acetic anhydride was added to the solution at room temperature with stirring and the mixture was further stirred for 18 hours. After the reaction mixture was poured into a saturated aqueous solution of sodium hydroxide to make it alkaline, the mixture was extracted twice with 100 ml of chloroform. The extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting residue was applied to a column of silica gel and eluted with chloroform / methanol (100: 1) to obtain 670 mg of the title compound in a colorless, amorphous form.

45 Reference Example 33 1- (O-Acetyl-N-benzyloxycarbonyl) tyrosyl-4- (3-phenylpropyl) homopiperazine.

In 2 ml of ethyl acetate, 670 mg of amorphous compound obtained in Reference Example 32 was dissolved and to the solution was added 7 ml of 3N hydrochloric acid in ethyl acetate at room temperature with stirring and the mixture was further stirred for 30 minutes, made alkaline with an aqueous solution of 50 sodium bicarbonate, saturated with hydrochloric acid and extracted twice with 100 ml of ethyl acetate. The extract was dried over magnesium sulfate and evaporated under reduced pressure to obtain 460 mg of 1- (O-acetyl-N-benzyloxycarbonyl) tyrosyl homopiperazine.

This compound was dissolved in 6 ml of dimethylformamide and to the solution were added 150 mg of potassium carbonate, 160 mg of sodium iodide and 210 mg of 1-bromo-3-phenylpropane at room temperature with stirring and the mixture was stirred for 20 hours. After addition of a saturated aqueous sodium chloride solution, the reaction mixture was extracted twice with 100 ml of chloroform and the extract was dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (100: 1) to obtain 430 mg of the title compound in a colorless, amorphous form.

Reference Example 34 1- (3-Phenylpropyl) -4-tyrosyl homopiperazine.

In 5 ml of methanol was dissolved 430 mg of amorphous compound obtained in Reference Example 33, to the solution was added 120 mg of potassium carbonate at room temperature with stirring and the reaction mixture was stirred for 70 hours. After adding a saturated aqueous solution of sodium chloride, the reaction mixture was acidified with citric acid and extracted twice with 100 ml of chloroform. The extract was dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (100: 1) to obtain 395 mg of 1- (N-benzyloxycarbonyl) tyrosyl-4 (3-phenylpropyl) homopiperazine.

This compound was dissolved in 15 ml of methanol and 0.05 ml of concentrated hydrochloric acid and 150 mg of 5% palladium on carbon were added to the solution while cooling with ice. After heating the reaction mixture to room temperature, the catalytic reduction was carried out in a hydrogen atmosphere for 8 hours. The palladium on carbon catalyst was filtered off with suction and washed with methanol. The filtrates were combined and evaporated to remove the solvent under reduced pressure and to the resulting residue was added a saturated aqueous solution of sodium chloride. The mixture was made alkaline with an aqueous solution of sodium bicarbonate, precipitated insoluble material was dissolved by adding a small amount of methanol and the mixture was extracted twice with 80 ml of chloroform. The extract was dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure and the resulting residue was applied to a column of silica gel and eluted with chloroform / methanol (20: 1) to obtain 180 mg of the title mentioned in the preamble. compound in a colorless, amorphous form.

Example CXXIV

1- [N- (5-Isoquinolinesulfonyl) tyrosyl] -4- (3-phenylpropyl) homopiperazine.

180 ml of amorphous compound obtained in Reference Example 34 were dissolved in 4 ml of tetrahydrofuran and 137 mg of 5-isoquinolinesulfonyl chloride.HCl and 0.2 ml of triethylamine were added to the solution at room temperature with stirring and the mixture was stirred for 15 hours. After adding a saturated aqueous solution of sodium chloride, the reaction mixture was made alkaline with sodium bicarbonate and the precipitated, insoluble material was made oily by adding a small amount of methanol and the mixture was extracted twice with 50 ml of chloroform. The extract was dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting residue was applied to a column of silica gel and a preparative plate for thin layer chromatography and eluted with chloroform / methanol (10: 1) to obtain 130 mg of the title compound in a colorless, amorphous form.

40

Reference Example 35 1- [N- (Tert-butoxycarbonyl) -p-nitrophenylalanyl] -4- (p-methoxyphenyl) piperazine.

In 120 ml of tetrahydrofuran, 100 ml of methylene chloride and 100 ml of chloroform, 9.00 g of N- (tert-butoxycarbonyl) -p-nitrophenylalanine were dissolved and the solution was successively added 45.68 g of N- (p-methoxyphenyl) piperazine dihydrochloride and 4.44 g of N-hydroxybenzotriazole monohydrate as well as 20 ml of triethylamine and 6 g of DCC and the mixture was stirred at room temperature for 18 hours. The reaction mixture was evaporated to one third of the original volume under reduced pressure and after the addition of 200 ml of 2.5% potassium carbonate as an aqueous solution, extracted twice with 200 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure 50 to crystallize a product which was washed with methanol to obtain 10.75 g of the title compound in the form of colorless crystals.

Reference Example 36 1- [N- (Tert-butoxycarbonyl) -p-aminophenylalanyl] -4- (p-methoxyphenyl) piperazine.

55 10.75 g of crystals obtained in Reference Example 35 were dissolved in a mixed solvent of 100 ml of tetrahydrofuran and 20 ml of methanol and 5 g of 5% palladium on carbon were added to the solution and the mixture was stirred at room temperature for 2 hours in room temperature. a hydrogen atmosphere 194549 34. After filtering off the insoluble material, the filtrate was evaporated under reduced pressure and the resulting residue was applied to a slurry gel column and eluted with chloroform / methanol (200: 1 to 100: 1) to obtain 10.08 g of the in the the above compound in a colorless, amorphous form.

5

Reference Example 37 1- [3- (p-Aminophenyl) -2- (tert-butoxycarbonylamino) propyl] -4- (p-methoxyphenyl) piperazine.

2.54 g of lithium aluminum hydride was suspended in 75 ml of tetrahydrofuran, a solution of 8.91 g of aluminum chloride in 75 ml of ether was added to the suspension while cooling with ice, and also a solution of 10.08 g of amorphous compound obtained in Reference Example 36 in 100 ml of tetrahydrofuran, was added dropwise for 20 minutes while cooling with ice. Under the same conditions, the mixture was stirred for one hour and after the reaction was terminated by adding a small amount of water, 70 ml of 30% potassium carbonate as an aqueous solution and 200 ml of chloroform were added to the reaction mixture, which was then filtered off for removal of the insoluble material using silica gel as a filter aid. The insoluble material was washed with 20% methanol in chloroform and the filtrates were combined and evaporated under reduced pressure. To the residue, 200 ml of saturated aqueous sodium bicarbonate solution was added, the mixture was extracted twice with 100 ml of chloroform, and the extract was dried over magnesium sulfate and evaporated under reduced pressure. The resulting residue was applied to a column of silica gel and eluted with chloroform / methanol (100: 1) to obtain 7.72 g of the title compound in a colorless, amorphous form.

Reference Example 38 1- [2- (Tert-butoxycarbonylamino) -3- (p-phthalimidephenyl) propyl] 4- (p-methoxyphenyl) piperazine.

7.0 g of amorphous compound obtained in Reference Example 37 were dissolved in 70 ml of chloroform, 2.66 g of phthalic anhydride was added to the solution. The mixture was refluxed for one hour, evaporated under reduced pressure and after addition of 100 ml toluene further refluxed for 2 hours. The solvent was evaporated under reduced pressure and the resulting residue was applied to a column of silica gel and eluted with chloroform / methanol (100: 1 to 50: 1) to obtain 8.91 g of the title compound in the title. form of colorless crystals.

Example CXXV

N-2- [4- (p-Methoxyphenyl) piperazinyl] -1- (p-phthalimide benzyl) ethyl 5-isoquinolinesulfonamide.

In 50 ml of ethyl acetate was dissolved 8.91 g of the crystals obtained in Reference Example 38, 100 ml of 4N hydrochloric acid in ethyl acetate was added to the solution and the mixture was stirred at room temperature for one hour. The reaction mixture was evaporated under reduced pressure and to the residue was added 200 ml of saturated aqueous solution of sodium bicarbonate and the mixture was extracted twice with 100 ml of 20% methanol / chloroform. The extract was dried over magnesium sulfate 40 and evaporated under reduced pressure to crystallize an amino-free compound. The crystals were suspended in 120 ml of tetrahydrofuran, to the suspension were added 5.0 g of 5-isoquinolinesulfonyl chloride.HCl and 20 ml of triethylamine while cooling with ice and after heating to room temperature, the mixture was stirred for 2 hours. After the addition of 200 ml of water, the crystals formed were collected. The filtrate was extracted twice with 100 ml of chloroform and the extract was dried over magnesium sulfate and evaporated to dryness under reduced pressure to obtain a residue. The residue with the crystals was washed successively with methanol, ethyl acetate and hexane to obtain 6.49 g of the title compound in the form of colorless crystals.

Example CXXVI

50 N- {2- [4- (p-Methoxyphenyl) piperazinyl] -1- (p-phthalimide benzyl) ethyl} -N-methyl-5-isoquinolinesulfonamide.

4.71 g of the crystals obtained in Example CXXV were dissolved in 70 ml of dimethylformamide, 500 mg of 60% sodium hydride and 1 ml of methyl iodide were successively added to the solution while cooling with ice and the mixture was stirred under the same conditions for 3 hours. The reaction was terminated by adding a small amount of water and after adding 200 ml of saturated aqueous ammonium chloride solution, the mixture was extracted twice with 100 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure. To the resulting residue were added 50 ml of acetic anhydride and 1.2 g of sodium acetate and the 194549 mixture was stirred at 80 ° C for one hour and then evaporated to dryness under reduced pressure and the resulting residue was dissolved in 200 ml of ethyl acetate. The solution was washed successively with 100 ml of saturated aqueous sodium bicarbonate solution and 100 ml of saturated aqueous sodium chloride solution, dried over magnesium sulfate and evaporated under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (100: 1) to obtain 4.84 g of the title compound in the form of colorless crystals.

Example CXXVII

1.5 g of crystals obtained in Example CXXV were suspended in 30 ml of ethanol, 3 ml of hydrozine hydrate was added to the suspension and the mixture was refluxed for one hour. After addition of 10% sodium hydroxide dissolved in water, the reaction mixture was extracted twice with 30 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure to form crystals which were washed with a mixed solvent of ethyl acetate and methanol to give 1.14 g of N- {1- (p-aminobenzyl) -2- [4- (p-methoxyphenyl) piperazinyl] ethyl} -5-isoquinolinesulfonamide as pale yellow crystals.

Example CXXVIII

4.64 g of crystals obtained in Example CXXVI were suspended in 80 ml of ethanol, 8 ml of hydrazine hydrate was added to the suspension and the mixture was refluxed for 90 minutes. After adding 100 ml of 10% sodium hydroxide, the reaction mixture was extracted twice with 80 ml of chloroform and the extract was dried over magnesium sulfate and evaporated under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (100: 1 to 50: 1) to give 3.29 g of N- {1- (p-aminobenzyl) -2- [4- (p -25 methoxyphenyl) piperazinyl] ethyl} -N-methyl-5-isoquinolinesulfonamide in a yellow, amorphous form.

Example CXXIX

500 mg of amorphous compound obtained in Example CXXVIII were dissolved in 5 ml of pyridine, 305 mg of 5-isoquinolinesulfonyl chloride.HCl was added to the solution while cooling with ice and the mixture was stirred under the same conditions for 20 minutes and then at room temperature for one hour . After adding 30 ml of a saturated aqueous solution of sodium bicarbonate, the reaction mixture was extracted twice with 20 ml of chloroform and the extract was dried over magnesium sulfate and evaporated under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (50: 1) to obtain 665 mg of N- {1- [p- (5-35 isoquinolinesulfonylaminobenzyl)] -2- [4- (p-) methoxyphenyl) piperazinyl] ethyl} -N-methyl-5-isoquinolinesulfonamide in a yellow, amorphous form.

Example CXXX

200 mg of crystals obtained in Example CXXVII were dissolved in 3 ml of pyridine, 40 130 mg of 5-isoquinoline sulfonyl chloride 1/2 sulfate was added to the solution while cooling with ice and the mixture was stirred with cooling on ice for 20 minutes and then at room temperature extracted twice with 20 ml of chloroform for one hour and after the addition of 30 ml of saturated aqueous solution of sodium bicarbonate. The extract was dried over magnesium sulfate and evaporated under reduced pressure and the resulting residue was applied to a silica gel column and eluted with chloroform / 45 methanol (50: 1 to 25: 1) to obtain 270 mg of N- {1- [p - (5-isoquinolinesulfonylaminobenzyl)] - 2- [4- (p-methoxyphenyl) piperazinyl] ethyl} -5-isoquinolinesulfonamide in a yellow amorphous form.

Example CXXXI

N - [[1- {p- [N-5-isoquinolinesulfonyl) -N- (methylamino) benzyl]} - 2- [4- (p-methoxyphenylpiperazinyl] ethyl]] - N-50 methyl-5-isoquinolinesulfonamide.

503 mg of amorphous compound obtained in Example CXXIX was dissolved in 8 ml of dimethylformamide, 50 mg of 60% sodium hydride and 0.1 ml of methyl iodide were added to the solution while cooling with ice and the mixture was stirred for 1 hour with cooling with ice. After adding 30 ml of saturated aqueous sodium chloride solution, the reaction mixture was extracted with 30 ml of ethyl acetate and the extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (100: 1) to obtain 488 mg of the compound mentioned in the 194549 36 in a yellow, amorphous form.

Example CXXXII

N- {1- [p- (4-Picolyloxy) benzyl] -2- [4- (2-pyrimidyl) piperazinyl] ethyl} -N-methyl-5-isoquinolinesulfonamide.

In 10 ml of a mixture of dried tetrahydrofuran / dried dimethylformamide (1: 1), 100 mg of amorphous compound obtained in Example XLII was dissolved, to the solution was added 34.8 mg of 4-picolyl chloride hydrochloride and then 24 mg of triethylamine and the mixture was added stirred at room temperature for 30 minutes. After the addition of 10 mg of 60% sodium hydride, the mixture was stirred overnight at room temperature and after the addition of 20 g of water was extracted twice with 20 ml of chloroform. The extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting residue was applied to a silica gel column and extracted with chloroform / methanol (100: 1) to obtain 73 mg of the title compound in a colorless, amorphous form.

15

Example CXXXIII

N- {1- [p- (2-Picolyloxy) benzyl] -2- [4- (2-pyrimidyl) piperazinyl] ethyl} -N-methyl-5-isoquinolinesulfonamide.

The same procedure as described in Example CXXXII was repeated, except that the same amount of 2-picolyl chloride hydrochloride was used instead of 4-picolyl chloride hydrochloride to obtain 74.4 mg of the title compound in a colorless, amorphous form.

Example CXXXIV

N- {1- [p- (4-Picolyloxy) benzyl] -2- [4- (2-pyridyl) piperazinyl] ethyl} -N-methyl-5-isoquinolinesulfonamide.

The same process as described in Example CXXXII was repeated, except that the product of Example XLVI was used instead of the amorphous compound obtained in Example XLII, to obtain the title compound in a colorless, amorphous form, with a yield of 53.5%.

Example CXXXV

N- {1- (p- [2-Picolyloxy) benzyl] -2- [4- (2-pyridyl) piperazinyl] ethyl} -N-methyl-5-isoquinolinesulfonamide.

The same process as described in Example CXXXIII was repeated except that the product of Example XLVI was used instead of the amorphous compound obtained in Example XLII to obtain the title compound in a colorless, amorphous form, with a yield of 59.5%.

Example CXXXVI

N- (2-Aminoethyl) -N- [2- (4-benzyloxycarbonyl-piperazinyl) -1- (p-methoxybenzyl) ethyl] -5-isoquinolinesulfonamide. 1.0 g of the product obtained in Example LXXIII was dissolved in 5 ml of tetrahydrofuran, 685 mg of triphenylphosphine and 340 mg of N-tert-butoxycarbonylethanolamine were added to the solution and then a solution of 530 mg of diisopropylazodicarboxylate in 3 ml of tetrahydrofuran was added dropwise stir and in an ice bath. After removal from the ice bath, the mixture was stirred at room temperature for 3 hours and poured into water and the mixture was made alkaline with sodium bicarbonate and extracted twice with 150 ml of chloroform. The extract was dried over magnesium sulfate and the solvent was evaporated under reduced pressure. The resulting oil was dissolved in 2 ml of ethyl acetate, 30 ml of 4N hydrochloric acid in ethyl acetate was added to the solution and the mixture was stirred at room temperature for 30 minutes. After adding 100 ml of 1N hydrochloric acid 45, the reaction mixture was washed twice with ethyl acetate and the aqueous layer was made alkaline with sodium bicarbonate and extracted twice with 150 ml of chloroform. The extract was dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure and the resulting oil was applied to a silica gel column and eluted with chloroform / methanol (100: 1 to 50: 1) to obtain 400 mg of the the above compound in a colorless, amorphous form.

50

Example CXXXVII

N- [2- (4-Benzyloxycarbonylpiperazinyl-1-p-methoxybenzyl) ethyl] -N- (2-dimethylaminoethyl) -5-isoquinolinesulfonamide.

6.08 g of the product obtained in Example LXXIII were dissolved in 30 ml of tetrahydrofuran, and 5.0 g of triphenylphosphine and 1.42 g of dimethyl ethanolamine were added to the 55 solution and then a solution of 3.21 was added dropwise g of diisopropyl azodicarboxylate in 10 ml of tetrahydrofuran with stirring in an ice bath. After removal from the ice bath, the reaction mixture was stirred at room temperature for 3 hours, diluted with ethyl acetate and extracted with 100 ml of 1N hydrochloric acid. The extract was made alkaline with sodium bicarbonate and extracted twice with 100 ml of chloroform and the extract was dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting oil was applied to a silica gel column and eluted with chloroform / methanol (200: 1 to 100: 1) to obtain 4.99 g of the title compound in a colorless, amorphous form.

Example CXXXVIII

N- (2-Acetoxyethyl) -N- [2- (4-benzyloxycarbonylpiperazinyl) -1 (p-methoxybenzyl) ethyl] -5-10 isoquinolinesulfonamide.

1.0 g of the product obtained in Example LXXIII was dissolved in 5 ml of tetrahydrofuran, 220 mg of ethylene glycol monoacetate and 685 mg of triphenylphosphine were added to the solution instead of N-tert-butoxycarbonylethanolamine according to the method described in Example CXXXVI to obtain 600 mg of the in the preamble in a colorless, amorphous form.

15

Example CXXXIX

N- [2- (4-Benzyloxycarbonyl-piperazinyl) -1- (p-methoxybenzyl) ethyl] -N- (2-hydroxyethyl) -5-isoquinolinesulfonamide.

In 6 ml of methanol and 3 ml of tetrahydrofuran was dissolved 600 mg of amorphous compound obtained in Example 20 CXXXVIII, to the solution was added 6 ml of 1N sodium hydroxide as an aqueous solution and the mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with water and extracted twice with 50 ml of chloroform and the extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting oil was applied to a silica gel column and eluted with chloroform / methanol (100: 1 to 50: 1) to obtain 403 mg of the title compound in a colorless, amorphous form.

Example CXL

N- {2- [4- (3,4-Dichlorobenzylamino) piperidino] -1- (p-methoxybenzyl) ethyl} -N-methyl-5-isoquinolinesulfonamide. The amorphous compound obtained in Example XCIV was subjected to alkaline hydrolysis, methylation with methyl iodide and potassium carbonate in dimethylformamide / tetrahydrofuran (1: 1) and refluxed with 3N hydrochloric acid to obtain N- {1- (p-methoxybenzyl) -2 - (4-oxopiperidino) ethyl} -N-methyl-5-oquinoline sulfonamide in a colorless, amorphous form.

3.30 g of this compound were dissolved in 30 ml of methanol, 1.89 g of 3.4-35 dichlorobenzylamine and 0.6 ml of acetic acid were added to the solution and the mixture was stirred at room temperature for 3 hours. The reaction mixture was cooled in an ice bath and, after adding 450 mg of sodium cyanoborium hydride, stirred while cooling with ice for 30 minutes and then at room temperature for one hour. This reaction mixture was made alkaline with sodium bicarbonate and extracted twice with 150 ml of chloroform and the extract was dried over magnesium sulfate and evaporated to remove the solvent under a reduced pressure. The resulting oil was applied to a silica gel column and eluted with chloroform / methanol (100: 1 to 50: 1) to obtain 2.78 g of the title compound in a colorless, amorphous form.

Example CXLI

45 N- [2- {4- [N- (3,4-Dichlorobenzyl) -N-methylamino] piperidino} -1 (p-methoxybenzyl) ethyl]] - N-methyl-5-isoquinolinesulfonamide.

In 10 ml of tetrahydrofuran and 10 ml of dimethylformamide was dissolved 1.62 g of amorphous compound obtained in Example CXL, to the solution was added 115 mg of 60% sodium hydride with stirring and cooling with ice and the mixture was reacted at the same temperature for 5 minutes and then 50 at room temperature for 15 minutes and again cooled with ice. After adding 405 mg of methyl iodide, the mixture was reacted at the same temperature for 5 minutes and then at room temperature for 2 hours and poured into water. The mixture was extracted with 200 ml of ethyl acetate and the extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting oil was applied to a silica gel column and eluted with chloroform / methanol (200: 1 to 100: 1) to give 880 mg of the title compound in a colorless, amorphous form.

4 194549 38

Reference Example 39 4-Chlorocinnamyl alcohol.

25.9 g of p-chloro-cinnamic acid (cinnamic acid will be referred to as cinnamic acid) were dissolved in 250 ml of methanol, 1.5 ml of concentrated sulfuric acid was added to the solution and the mixture was heated under reflux for 2 hours. The reaction mixture was poured onto ice and the mixture was made alkaline with sodium bicarbonate and extracted twice with 1000 ml of chloroform. The extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure, and the resulting residue was applied to a silica gel column and eluted with hexane / 10 ethyl acetate (10: 1) ter obtaining 26.5 g of methyl p-chloro cinnamate.

This compound was dissolved in 250 ml of toluene, to the solution was added 200 ml of 1.5M diisobutylaluminum hydride in toluene with stirring and cooling with ice and the mixture was stirred for 2 hours. The reaction mixture was poured into ice, acidified with concentrated hydrochloric acid and extracted twice with 700 ml of benzene. The extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure, and the resulting residue was applied to a silica gel column and eluted with hexane / ethyl acetate (4: 1) ter obtaining 21.0 g of the title compound as colorless crystals.

Reference Example 40 N-4-Chlorocinnamyl-1,2-phenylenediamine.

11.9 g of crystals obtained in Reference Example 39 were dissolved in 120 ml of chloroform, 10.1 g of thionyl chloride was added to the solution with stirring in an ice bath and after removal from the ice bath, the mixture was stirred for one hour while the reaction temperature was being adjusted increased to room temperature. Chloroform and an excess of thionyl chloride were evaporated under reduced pressure, benzene was added to the residue and the solvent was evaporated under reduced pressure. The resulting residue was applied to a silica gel column and eluted with hexane / ethyl acetate (15: 1) to obtain 11.3 g of 4-chloro-cinnamyl chloride as colorless crystals.

19.6 g of 1,2-phenylenediamine were dissolved in 300 ml of dimethylformamide, 11.3 g of the above 4-chloro-cinnamyl chloride in crystal form and 12.5 g of potassium carbonate were added to the solution at room temperature with stirring and the mixture was stirred for 48 minutes. stirred under the same conditions for hours. After addition of water and sodium chloride, the reaction mixture was extracted twice with 1000 ml of chloroform and the extract was dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting residue was applied to a column of silica gel and eluted with hexane / ethyl acetate (3: 1) to obtain 12.85 g of the title compound as colorless crystals.

Example CXLII

N- [2- (4-Chlorocinnamylamino) phenyl] -5-isoquinolinesulfonamide.

40 12.85 g of crystals obtained in Reference Example 40 were dissolved in 200 ml of pyridine, 15.1 g of 5-isoquinolinesulfonyl chloride hydrochloride was added to the solution with stirring in an ice bath and after removal from the ice bath the mixture was reacted at room temperature for 18 hours.

The reaction mixture was poured onto ice, made alkaline with sodium bicarbonate and extracted twice with 1000 ml of chloroform. The extract was dried over magnesium sulfate and the solvent was evaporated under reduced pressure to form hardly soluble crystals. Chloroform was added to the crystals, the whole was refluxed and then cooled, and the resulting crystals were collected by suction filtration, washed with chloroform and dried under reduced pressure to obtain 17.23 g of the title compound as colorless crystals.

50

Example CXLIII

N- [2- (4-Chlorocinnamylamino) phenyl] -N-methyl-5-isoquinolinesulfonamide.

380 mg of crystals obtained in Example CXLII were dissolved in 6 ml of methanol, 10 ml of a solution of diazomethane in ether was added at room temperature with stirring and the mixture was stirred for 18 hours. The solvent was evaporated under reduced pressure to obtain an oil which was then applied to a silica gel column and eluted with hexane / ethyl acetate (1: 1) to obtain acetate, which was then recrystallized from hexane / 39 194549

ethyl acetate to obtain 270 mg of the title compound as colorless crystals. Example CXLIV

1 - (4-Chlorocinnamyl) -4- (5-isoquinolinesulfonyl) -1,2,3,4t etrahydroquinoxaline.

5.0 g of crystals obtained in Example CXLII were dissolved in 75 ml of dimethylformamide, 4.6 g of potassium carbonate and 2.19 g of 1,2-dibromoethane were added to the solution with stirring and the mixture was stirred for 60 hours . The reaction mixture was poured into water, saturated with sodium chloride, and extracted twice with 400 ml of chloroform. The extract was dried over magnesium sulfate, evaporated to remove the solvent under reduced pressure.

The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (400: 1) and then with hexane / ethyl acetate (2: 1) to obtain 3.32 g of the title compound in a yellow , amorphous form.

Example CXLV

The same process as described in Example CXLII was repeated, except that N- [3- (3-pyridyl) allyl] -1,2-phenylenediamine was used instead of N- (4-chloro-cinnamyl) -1,2-phenylenediamine, to give N- {2- [3- (3-pyridyl) allylamino] phenyl} -5-isoquinolinesulfonamide in a brown, amorphous form.

Example CXLVI

The amorphous compound obtained in Example CXLV was treated according to the method described in Example CXLIII to obtain N- {2- [3- (3-pyridyl) allylamino) phenyl} -N-methyl-5-isoquinolinesulfonamide.

Reference Example 41 2- Amino-3- (4-chloro-cinnamylamino) pyridine.

7.71 g of p-chloro-cinnamyl chloride and 13.5 g of 2,3-diaminopyridine were dissolved in 220 ml of dimethylformamide and 8.6 g of potassium carbonate were added to the solution and the mixture was stirred at room temperature for 50 hours and after addition of 300 ml of water extracted twice with 200 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure and the resulting residue was applied to a silica gel column and eluted with chloroform / methanol (100: 1 to 50: 1) to obtain 4.52 g of the title. said compound in the form of yellow crystals.

Example CXLVII

3- (4-Chlorocinnamylamino) -2- (5-isoquinolinesulfonylamino) pyridine.

In 50 ml of pyridine, 4.52 g of the crystals obtained in Reference Example 41 were dissolved, 5.8 g of 5-isoquinolinesulfonyl chloride hydrochloride and 3 g of dimethylaminopyridine were added to the solution and the mixture was stirred at room temperature for 18 hours after the adding 150 ml of water, extracted twice with 80 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure and the resulting residue was applied to a silica gel column and eluted with chloroform / methanol (100: 1) and the resulting crystals were washed with ethyl acetate to obtain 1.2 g of the compound mentioned in the preamble as yellow crystals.

Reference example 42

Methyl 4-amino-3- (4-chloro-cinnamylamino) benzoate.

45 In 40 ml of dimethylformamide, 5.0 g of methyl 3,4-diaminobenzoate was dissolved and 2.07 g of potassium carbonate and 1.87 g of p-chloro-cinnamyl chloride were added to the solution and the reaction was carried out according to the method described in Reference Example 40 to obtain of 2.0 g of the title compound in the form of a light brown oil.

50 Example CXLVIII

Methyl 4- (5-isoquinolinesulfonamino) -3- (4-chloro-cinnamylamino) benzoate.

In 18 ml of pyridine was dissolved 1.8 g of oil obtained in Reference Example 42, 1.29 g of 5-isoquinolinesulfonyl chloride hydrochloride was added to the solution with stirring and cooling with ice and the mixture was treated according to the method described in Example CXLII to obtain of a residue 55 which was then applied to a silica gel column and eluted with chloroform / methanol (100: 1) to give 1.28 g of the title compound in the form of light yellow crystals.

194549 40

Reference Example 43 N-cinnamyl-1,2-phenylenediamine.

3.24 g of o-phenylenediamine was dissolved in 30 ml of dimethylformamide, 2.07 g of potassium carbonate and 1.52 g of cinnamyl chloride were added to the solution and the mixture was stirred at room temperature overnight. After adding 100 ml of water, the reaction mixture was extracted twice with 100 ml and 50 ml of chloroform and the extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform to give 2.0 g of the title compound as light brown crystals.

10

Example CIL

N- (2-cinnamulamino) phenyl-5-isoquinolinesulfonamide.

1.8 g of crystals obtained in Reference Example 43 were dissolved in 18 ml of pyridine, 1.83 g of isoquinoline sulfonyl chloride hydrochloride was added to the solution and the mixture was stirred at room temperature for 18 hours. After adding 50 ml of water, the reaction mixture was extracted twice with 80 ml of chloroform and the extract was washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (100: 1) to obtain 2.40 g of the title compound as light red crystals.

Reference Example 44 N- (4-Chlorocinnamyl) -1,3-phenylenediamine.

3.24 g of m-phenylenediamine was dissolved in 40 ml of dimethylformamide, 2.07 g of potassium carbonate and 1.87 g of p-chloro-cinnamyl chloride were added to the solution and the mixture was subjected to a reaction according to the method described in Reference Example 40. The resulting residue was applied to a silica gel column and eluted with n-hexane / ethyl acetate (3: 1 to 2: 1) to give 1.70 g of the title compound as a light brown oil.

Example CL

N- [3- (4-Chlorocinnamylamino) phenyl] -5-isoquinolinesulfonamide.

1.7 g of oil obtained in Reference Example 44 were dissolved in 18 ml of pyridine, 1.99 g of 5-isoquinolinesulfonyl chloride.HCl was added to the solution with stirring and cooling with ice, and the same procedure as described in Example CXLII was repeated to obtain 1 45 g of the compound mentioned in the preamble 35 in the form of a light brown oil.

Example CU

N- {2- (p-Chlorocinnamylamino) phenyl} -N- (2-hydroxyethyl) -5-isoquinolinesulfonamide.

1.5 g of crystals obtained in Example CXLII were dissolved in 8 ml of tetrahydrofuran, 1.32 g of triphenylphosphine and 420 mg of ethylene glycol monoacetate were added to the solution 40 and a solution of 1.01 g of diisopropylazodicarboxylate in 2 ml of tetrahydrofuran was added dropwise with stirring in an ice bath. After being removed from the ice bath, the mixture was warmed to room temperature, stirred for 3 hours, diluted with ethyl acetate, and extracted twice with 70 ml of 2N hydrochloric acid. The aqueous layer was made alkaline with sodium bicarbonate and extracted twice with 150 ml of chloroform and the extract was dried over magnesium sulfate and evaporated under reduced pressure to remove the solvent. The oily residue obtained was dissolved in 20 ml of methanol and 20 ml of tetrahydrofuran, 20 ml of 1 N sodium hydroxide dissolved in water was added to the solution and the reaction was carried out at room temperature for 2 hours. The reaction mixture was diluted with water and extracted twice with 100 ml and 50 ml of chloroform and the extract was dried over 50 magnesium chloride and evaporated under reduced pressure to remove the solvent. The resulting oil was applied to a silica gel column and eluted with chloroform / methanol (100: 1 to 50: 1) to obtain 1.59 g of the title compound in a yellow, amorphous form.

Example CLII

55 N-2- (p-Chlorocinnamylamino) phenyl-N- (2-dimethylaminoethyl) 5-isoquinolinesulfonamide.

2.0 g of the crystals obtained in Example CXLII were dissolved in 10 ml of tetrahydrofuran, 1.75 g of triphenylphosphine and 520 mg of N, N-dimethylethanolamine were added to the solution, and a solution of 1.3 g was added dropwise thereto to 41 194549 diisopropylazodicarboxylate in 3 ml of tetrahydrofuran with stirring in an ice bath. After being removed from the ice bath, the mixture was warmed to room temperature, stirred for 3 hours and then diluted with ethyl acetate and extracted twice with 100 ml of 2N hydrochloric acid. The extract was made alkaline with an aqueous solution of sodium bicarbonate and extracted twice with 200 ml of chloroform. The extract was dried over magnesium sulfate and evaporated under reduced pressure to remove the solvent and a resulting oil was applied to a silica gel column and eluted with chloroform / methanol (100: 1) to obtain 1.35 g of the title. said compound in a yellow, amorphous form.

Examples CLIII to CLXXI

In Examples CLIII to CLXXI, the following general reaction was used as indicated on the formula sheet as reaction (a).

Example CLIII

15 (n = 2, Aryl = 4-chlorophenyl).

N- [2- (4-Chloro-α-methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.

7.30 g of N- (2-aminoethyl) -5-isoquinolinesulfonamide were dissolved in 150 ml of methanol, 6.30 g of p-chlorobenzalacetone was added to the solution and the mixture was stirred at room temperature for 36 hours. After adding 1.32 g of sodium tetrahydride borate while cooling with ice water, the mixture was stirred for 30 minutes. The reaction mixture was evaporated to half the original volume under reduced pressure and washed three times with water after adding 300 ml of ethyl acetate. The aqueous layer was extracted with 100 ml of ethyl acetate and the extract was washed with water as indicated above. The ethyl acetate layers were combined, washed twice with saturated aqueous sodium chloride solution, dried over magnesium sulfate, filtered, and evaporated under reduced pressure to remove the solvent. The resulting residue was purified using a silica gel column (silica gel: 200 g; eluent: 5% methanol in chloroform) to obtain 6.78 g of the title compound in a colorless, amorphous form, with the remaining starting material was recovered.

Example CLIV (n = 2, Aryl = phenyl).

N- [2- (α-Methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Colorless amorphous form;

Example CLV

(n-2, Aryl = 2,4-difluorophenyl).

N- [2- (2,4-Difluoro-α-methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Colorless amorphous form.

40 Example CLVI

(n = 2, Aryl-2,4-dichlorophenyl).

N- [2- (2,4-Dichloro-α-methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Colorless amorphous form.

45 Example CLVII

(n = 2, Aryl = 3-chlorophenyl).

N- [2- (3-Chloro-α-methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Colorless amorphous form.

Example CLVIII

(n = 2, Aryl-2-nitrophenyl).

N- [2- (α-Methyl-2-nitrocinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Light yellow, amorphous form.

55 Example CLIX

(n = 2, Aryl = 4-nitrophenyl).

N- [2- (α-Methyl-4-nitrocinnamylamino) ethyl] -5-isoquinoline sulfonylamide.

194549 42

Light yellow, amorphous form.

Example CLX

(n = 2, Aryl = 4-methylphenyl).

5 N- [2- (a, 4-Dimethylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Colorless amorphous form.

Example CLXI

(n = 2, Aryl = 3,4-methylenedioxyphenyl).

N- [2- (α-Methyl-3,4-methylenedioxycinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Colorless amorphous form.

Example CLXII (n = 2, Aryl = 2-pyridyl).

N- {2- [1-Methyl-3- (2-pyridyl) -2-propenylamino] ethyl} -5-isoquinolinesulfonamide.

Example CLXIII (n = 2, Aryl = 4-pyridyl).

N- {2- [1-Methyl-3- (4-pyridyl) -2-propenylamino] ethyl} -5-isoquinolinesulfonylamide. 20 Colorless amorphous form.

Example CLXIV (n = 2, Aryl = 2-thienyl).

N- {2- [1-Methyl-3- (2-thienyl) -2-propenylamino] ethyl} -5-isoquinolinesulfonamide.

25 Colorless amorphous form.

Example CLXV (n = 2, Aryl = 2-furyl).

N- {2- [3- (2-Furyl) -1-methyl-2-propenylamino] ethyl} -5-isoquinolinesulfonamide.

30 Colorless amorphous form.

Example CLXVI

(n = 2, Aryl = 4-fluorophenyl).

N- [2- (4-Fluoro-α-methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.

35 Colorless amorphous form.

Example CLXVII

(n = 2, Aryl = 4-bromophenyl).

N- [2- (4-Bromo-α-methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.

40 Colorless amorphous form.

Example CLXVIII

(n = 2, Aryl = 4-isopropylphenyl).

N- [2- (4-Isopropyl-α-methyl-cinnamylamino) -ethyl] -5-isoquinolinesulfonamide.

45 Colorless amorphous form.

Example CLXIX

(n = 2, Aryl = 4-methoxyphenyl).

N- [2- (4-Methoxy-α-methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.

50 Colorless amorphous form.

Example CLXX

(n = 2, Aryl = 4-hydroxyphenyl).

N- [2- (4-hydroxy-α-methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.

55 Colorless crystals.

43 194549

Example CLXXI (n = 3, Aryl = phenyl).

N- [3- (α-Methylcinnamylamino) propyl] -5-isoquinormesulfonamide.

Colorless amorphous form.

5

Examples CLXXII to CLXXXVIII

In the examples CLXXII to CLXXXVIII, the following general reaction was applied as indicated on the formula sheet as reaction (b).

Example CLXXII

(n = 2, m = 1, Aryl = 4-chlorophenyl).

N- [2- (4-Chlorocinnamylamino) ethyl] -5-isoquinolinesulfonamide.

2.01 g of N- (2-aminoethyl) -5-isoquinolinesulfonamide were dissolved in 30 ml of methanol, 1.60 g of p-chloro-cinnamaldehyde was added to the solution and the mixture was stirred for one hour at room temperature. After an addition of 350 mg of sodium tetrahydride borate in parts while cooling with ice, the mixture was stirred for 30 minutes. After addition of ethyl acetate, the reaction mixture was washed three times in succession with water and then twice with a saturated aqueous solution of sodium chloride and dried over magnesium sulfate. The mixture was filtered off and evaporated to remove the solvent under reduced pressure. A residue was purified using a column with silica gel (silica gel 80 g, eluent: 5% methanol in chloroform) and the resulting crystals were washed with benzene / hexane (1: 1) to obtain 2.30 g of the in the compound mentioned as colorless crystals.

Example CLXXIII (n = 2, m = 1, Aryl = phenyl).

N- (2-Cinnamylaminoethyl) -5-isoquinolinesulfonamide.

Colorless amorphous form.

Example CLXXIV

30 (n = 2, m = 1, Aryl = 4-dimethylaminophenyl).

N- [2- (4-Dimethylaminocinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Colorless amorphous form.

Example CLXXV

35 (n = 2, m = 1, Aryl = 4-fluorophenyl).

N- [2- (4-Fluoro-cinnamylamino) -ethyl] -5-isoquinolinesulfonamide.

Colorless amorphous form.

Example CLXXVI

40 (n = 2, m = 1, Aryl = 4-bromophenyl).

N- [2- (4-Bromophenylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Colorless crystals.

Example CLXXVII

45 (n = 2, m = 1, Aryl = 4-isopropylphenyl).

N- [2- (4-Isopropyl-cinnamylamino) -ethyl] -5-isoquinolinesulfonamide.

Colorless amorphous form.

Example CLXXVIII

50 (n = 2, m = 1, Aryl = 4-methoxyphenyl).

N- [2- (4-Methoxycinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Colorless crystals.

Example CLXXIX

55 (n = 2, m = 1, Aryl = 4-trifluoromethylphenyl).

N- [2- (4-Trifluoromethylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Colorless amorphous form.

194549 44

Example CLXXX

(n = 2, m = 2, Aryl = 4-trifluoromethylphenyl).

N- {2- [5- (4-Trifluoromethylphenyl) -2,4-pentadienylamino] ethyl} 5-isoquinolinesulfonamide.

Light yellow amorphous form.

5

Example CLXXXI

(n = 2, m = 3, Aryl = 4-trifluoromethylphenyl).

N- {2- [7- (4-Trifluoromethylphenyl) -2,416-heptatrienylamino] ethyl} -5-isoquinolinesulfonamide.

Light yellow amorphous form.

10

Example CLXXXII

[n = 2, m = 1, Aryl = 4- (2-methoxyethoxy) methoxyphenyl] N- {2- [4- (2-methoxyethhoxy) methoxycinnamylamino] ethyl} -5-isoquinolinesulfonamide. Colorless, amorphous form.

Example CLXXXIII

(n = 2, m = 1, Aryl = 4-hydroxyphenyl).

N- [2- (4-Hydroxy cinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Colorless crystals.

Example CLXXXIV

(n = 2, m = 1, Aryl = 1-naphthyl).

N- {2- [3- (1-Naphthyl) -2-propenylamino] ethyl} -5-isoquinoline sulfonamide.

Colorless crystals.

Example CLXXXV

(n = 2, m = 1, Aryl = 3,4,5-trimethoxyphenyl).

N- [2- (3,4,5-Trimethoxycinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Colorless, amorphous form.

Example CLXXXVI

(n = 2, m = 1, Aryl = 4-methoxycarbonylphenyl).

N- [2- (4-Carbomethoxycinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Colorless crystals.

Example CLXXXVH

(n = 2, m = 1, Aryl = 4-carboxyphenyl).

N- [2- (4-Carboxycinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Colorless crystals.

Example CLXXXVIII (n = 3, m = 1, Aryl = phenyl).

N- (3-cinnamylaminopropyl) -5-isoquinolinesulfonamide.

Colorless amorphous form.

45 Example CLXXXIX

N * {2- [3- (4-Chlorophenyl) -2-propynylamino] ethyl} -5-isoquinolinesulfonamide (compound 189-1); N- {2- [Bis- (3 - ((4-chlorophenyl)) -2-propynyl) amino] ethyl} -5-isoquinolinesulfonamide (compound 189-II); N- [3- (4-Chlorophenyl) -2-propynyl] -N- [2- (p-chlorophenyl) -2β-pyropylamino] ethyl] -5-isoquinolinesulfonamide (compound 189-111).

The reaction takes place according to the reaction scheme (c), as indicated on the formula sheet.

1.90 g of N- (2-aminoethyl) -5-isoquinolinesulfonamide and 1.39 g of 3-p-chlorophenyl-2-propynyl chloride were dissolved in 10 ml of dimethylformamide, 1.38 g of potassium carbonate was added to the solution and the mixture was added. stirred at room temperature for 24 hours. The reaction mixture was poured into 100 ml of ethyl acetate, washed three times with water and then twice with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate, filtered and evaporated to remove the solvent under reduced pressure. The resulting residue was separated and purified using a silica gel column (silica gel 100 g; eluent: 5% methanol / chloroform). By 194549 crystallization from a mixture of ether-hexane, 855 mg of compound 189-1 were obtained as colorless crystals, 220 mg of compound 189-11 and 214 mg of compound 189-111 in colorless, amorphous form.

Compound 189-1 Colorless crystals.

5 Compound 189-11

Colorless amorphous form.

Compound 189-111 Colorless amorphous form.

Example CXC

N- [2- (4-Chloro-N-methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.

The reaction is shown on the formula sheet as reaction (d).

1.50 g of the product from example CLXXII was dissolved in 10 ml of chloroform, 3 ml of methyl iodide was added to the solution at room temperature and the mixture was stirred for 40 minutes.

An excess of methyl iodide was immediately evaporated under reduced pressure and the resulting residue was purified on a silica gel column (silica gel 50 g, eluent: 5% methanol in chloroform) to obtain 720 mg of the title compound in a colorless , amorphous form.

Example CXCl 2 1- (Chlorocinnamyl) -4- (5-isoquinolinesulfonyl) piperazine.

The relevant reaction is as reaction (e), shown on the formula sheet.

1.31 g of the product from Example CLXXII were dissolved in 3 ml of dimethylformamide, 644 mg of 1,2-dibromoethane and 1.13 g of anhydrous potassium carbonate were added to the solution at room temperature and the mixture was stirred for 24 hours. After the addition of 100 ml of ethyl acetate, the ethyl acetate-containing layer was successively washed with water and a saturated aqueous solution of sodium chloride, which washing was carried out twice and dried over magnesium sulfate. The solution was filtered off and evaporated under reduced pressure and the resulting residue was purified on a silica gel column (silica gel. 50 g, eluent: 5% methanol in chloroform) to obtain 356 mg of the title compound in a colorless , amorphous form, while the remaining starting material was recovered.

Example CXCII

N-Ethyl-N- [2- (4-chloro-N-ethylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.

The reaction that occurs here is shown as reaction (f) on the formula sheet.

The process as indicated in Example CXCl was repeated except that 1.31 g of the product from Example CLXXII and 2.14 g of ethyl iodide were used as N-alkylating agent to obtain 720 mg of the title compound in a colorless , amorphous form.

Example CXCIII

40 N- [2- (4-Chloro-N-formylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.

The reaction that occurs is shown as reaction (g) on the formula sheet.

3 ml of formic acid and 3 ml of acetic anhydride were mixed with stirring at room temperature, and 1.41 g of the product from example CLXXII was added to the mixture and the mixture was stirred for one hour. The reaction mixture was added to 50 ml of ethyl acetate and 30 ml of saturated, 45 aqueous solution of sodium carbonate while cooling with ice and the mixture was stirred and after the foaming was terminated, the ethyl acetate-containing layer was washed twice with water and once with a saturated , aqueous solution of sodium chloride and dried over magnesium sulfate, filtered and evaporated under reduced pressure. The resulting residue was purified by column chromatography on silica gel (silica gel: 60 g, eluent: 2% methanol in chloroform) to obtain 1.49 g of the title compound as a mixture of two isomers (3: 2) in a colorless, amorphous form.

Example CXCIV

N- {2- [4-Chloro-N- (4-hydroxybenzyl) cinnamylamino] ethyl} -5-isoquinolinesulfonamide.

55 In 10 ml of methanol were dissolved 0.2 g of the product from example CLXXII and 0.13 g of p-hydroxybenzaldehyde, 60 mg of sodium cyanoborium hydride and two drops of acetic acid were added to the solution and the mixture was stirred at room temperature for two days. The reaction mixture 194549 46 was evaporated under reduced pressure and after addition of a saturated aqueous solution of sodium chloride extracted three times with 20 ml of ethyl acetate. The extracts were combined, washed with sodium chloride, dried over magnesium sulfate, filtered and evaporated under reduced pressure. The resulting residue was purified on a silica gel column (silica gel: 10 g, eluent: 2% methanol in chloroform), to obtain 150 mg of the title compound in a colorless, amorphous form.

Example CXCV

2-Methyl-5 - {[2- (4-chloro-N, N-dimethylcinnamylammonium) ethyl] aminosulfonyl} isoquinoline diodide.

83 mg of the product from example CLXXII was dissolved in 2.0 ml of dimethylformamide, 1.0 ml of methyl iodide was added to the solution and the mixture was stirred at room temperature for four hours. The excess of methyl iodide and dimethylformamide was evaporated under reduced pressure and the resulting residue was recrystallized from 5 ml of a methanol / chloroform mixture (1: 5). The crude crystals thus obtained were recrystallized from 10 ml of a methanol / chloroform (1: 5) mixture to obtain 78 ml of the title compound in the form of light yellow crystals.

Example CXCVI

2-Methyl-5- {N-methyl-N- [2- (4-chloro-N, N-dimethylcinnamylammonium) ethyl] amino sulfonyl} isoquinoline iodide. 83 mg of the product from example CLXXII were dissolved in 2.0 ml of dimethylformamide, 1.0 ml of methyl iodide and 83 mg of anhydrous sodium carbonate were added to the solution and the mixture was stirred at room temperature for four hours. The excess methyl iodide and dimethylformamide were evaporated under reduced pressure and after adding 10 ml of a methanol / chloroform mixture (1: 5), the mixture was stirred and then filtered to remove the insoluble material. The filtrate was evaporated under reduced pressure and to the concentrate was added 10 ml of a methanol / chloroform mixture (1: 5) to precipitate an insoluble material which was then filtered off. The evaporation and filtering was repeated twice to obtain 145 mg of the title compound in a yellow, amorphous form.

The isoquinoline compounds other than those from example CLXXIV can be treated with an excessive amount of methyl iodide in dimethylformamide as described in this example, to obtain the corresponding compounds, wherein the nitrogen atom on the isoquinoline ring is methylated to a quaternary nitrogen atom.

Example CXCVII

N- [2- (4-Chlorocinnamylamino) ethyl] -5-isoquinolinesulfonamide dihydrochloride.

2.00 g of the product from example CLXXII was suspended in 20 ml of methanol, the suspension was converted to a clear solution by adding 1 ml of concentrated hydrochloric acid and stirred for 10 minutes while cooling with ice to form crystals. The crystals were collected by filtration and recrystallized from a mixture of 20 ml of methanol and 3 ml of water to obtain 1.65 g of 40 of the corresponding dihydrochloride in the form of colorless crystals.

Example CXCVIII

N- [2- (α-Methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide dihydrochloride.

The same process as described in Example CXCIV was repeated, except that the product from Example CLIV was used as a starting material to obtain the corresponding dihydrochloride in the form of colorless crystals.

Example CIC

N- [2- (4-Chloro-α-methylcinnamylamino) ethyl] -5-isoquinoline sulfonamide dihydrochloride.

The same process as described in Example CXCIV was repeated, except that the product from Example CLIII was used as a starting material to obtain the corresponding dihydrochloride in the form of a white hygroscope powder.

Example CC

55 N- [2- (4-Bromo-cinnamylamino) -ethyl] -5-isoquinolinesulfonamide dihydrochloride.

The same process as described in Example CXCIV was repeated, except that the product from Example CLXXVI was used as the starting material to obtain the corresponding dihydro-1945 194549 in the form of colorless crystals.

Example CCI

N- (2-Cinnamylaminoethyl) -5-isoquinolinesulfonamide 1/2-fumarate.

303 mg of the product from example CLXXIII was dissolved in 5 ml of ethyl acetate, to the solution was added a solution of 89 ml of fumaric acid in 2 ml of methanol at room temperature and the mixture was stirred for 30 minutes to form crystals, which were then collected by filtration and washed with ethyl acetate to give 312 mg of the corresponding 1/2 fumarate in the form of colorless crystals.

10

Example CCII

N- [2- (α-Methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide 1/2 fumarate.

The same process as described in example CXCVIII was repeated, except that the product from example CLIV was used as a starting material to obtain the corresponding 1/2 fumarate in the form of colorless crystals.

Example CCIII

N- [2- (4-Chloro-α-methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide L - (+) - tartrate.

In 50 ml of ethyl acetate was dissolved 6.60 g of the product from example CLIII, to the solution was added a solution of 238 g of L - (+) - tartaric acid in methanol to form crystals, which were then collected by filtration and washed with ethyl acetate to give the corresponding L - (+) - tartrate as colorless crystals.

Example CCIV

N- (2-Aminoethyl) -N- (2-cinnamylaminoethyl) -5-isoquinoline sulfonamide trihydrochloride.

To a solution of 1.10 g of amorphous compound obtained in Example CLXXIII, 1.18 g of triphenylphosphine and 0.73 g of 2- (tert-butoxycarbonylamino) ethanol in 15 ml of tetrahydrofuran was added dropwise a solution of 0.78 g of diethyl azodicarboxylate in 5 ml of tetrahydrofuran while cooling with ice and the mixture was stirred at room temperature for 4 hours.

After re-cooling with ice, 0.39 g of triphenylphosphine was added to the reaction mixture and a solution of 0.26 g of diethyl azodicarboxylate in 3 ml of tetrahydrofuran was added dropwise and the reaction mixture was stirred at room temperature for one hour. The reaction mixture was evaporated under reduced pressure and the resulting residue was applied to a silica gel column and eluted with chloroform / methanol (19: 1), to obtain 0.99 g of a light orange, amorphous product. The product was dissolved in 20 ml of methanol, 7.7 ml of 4N hydrochloric acid in ethyl acetate was added to the solution and the mixture was stirred at room temperature for 3 hours. The reaction mixture was evaporated to remove the solvent under reduced pressure and to this was added ethyl acetate to form a solid. The solid was collected by filtration, washed with ethyl acetate and n-hexane and dried under reduced pressure to obtain 0.97 g of the title compound in the form of a colorless, hygroscopic powder.

Example CCV

N- (4-Aminobutyl) -N * [2- (4-chloro-cinnamylamino) -ethyl] -5-isoquinoline sulfonamide trihydrochlorides.

To a solution of 0.4 g of crystals obtained in Example CLXXII, 0.226 g of 4- (tert-45 butoxycarbonylaminojbutanol and 0.445 g of triphenylphosphine in 5 ml of tetrahydrofuran, was added a solution of 0.295 g of diethyl azodicarboxylate in 2 ml of tetrahydrofuran with stirring and cooling with ice .

The mixture was stored at room temperature and evaporated under reduced pressure to obtain a residue which was then applied to a silica gel column and eluted with methanol / chloroform (2:98) to obtain 0.28 g of oil. To a solution of the oil in 1 ml of methanol was added 4N hydrochloric acid / 50 ethyl acetate to form a precipitate, which was then collected by filtration, washed with ethyl acetate and dried to give 0.2 g of the title compound as a colorless powder.

Example CCVI

55 N- [2- (4-Chloro-N-methylcinnamylamino) ethyl] -N- [2- (4-piperidyl) ethyl] -5-isoquinolinesulfonamide.

To a solution of 0.39 g of amorphous compound obtained in Example CXC, 0.145 g of 4-piperidinethanol and 0.255 g of triphenylphosphine in 5 ml of tetrahydrofuran was added a solution of 0.245 g of diethylazodi-194549 48 carboxylate in 2 ml of tetrahydrofuran while cooling with ice and cooling. The mixture was stirred at room temperature for 1 hour and evaporated to remove the solvent under reduced pressure. After adding 30 ml of ethyl acetate, the mixture was extracted three times with 5 ml of 1N hydrochloric acid. The extract was made alkaline with sodium bicarbonate and extracted three times with 10 ml of ethyl acetate.

The organic layer was dried over magnesium sulfate and evaporated to remove the solvent. The resulting residue was applied to a chromatographic column of alumina and eluted with 1% methanol in chloroform to give 180 mg of the title compound as a colorless oil.

The oil thus obtained was dissolved in 1 ml of methanol and to this was added 0.3 ml of 4N hydrochloric acid in ethyl acetate and then 30 ml of ether to obtain the corresponding trihydrochloride as a colorless powder.

Example CCVII

N- [2- (4-Chloro-N-methylcinnamylamino) ethyl] -N- [2-morpholinoethyl) -5-isoquinoline sulfonamide trihydrochloride. To a solution of 1 g of amorphous compound obtained in Example CXC, 0.377 g of 2-N-morpholinoethanol and 1.25 g of triphenylphosphine in 5 ml of tetrahydrofuran was added dropwise a solution of 0.835 g of diethyl azodicarboxylate in 2 ml of tetrahydrofuran with stirring and cooling with ice and the mixture was stirred for 2 hours. The solvent was removed by evaporation under reduced pressure, to the residue was added 20 ml of ethyl acetate and the mixture was extracted three times with 10 ml of 1N hydrochloric acid. The extract was made alkaline with sodium bicarbonate and extracted three times with 10 ml of ethyl acetate. The extract was dried over magnesium sulfate and evaporated under reduced pressure to remove the solvent. The resulting residue was applied to a silica gel column and eluted with methanol / ethyl acetate (10:90) to obtain an oil.

The oil thus obtained was dissolved in 4 ml of methanol and added in 2 ml of 4N hydrochloric acid in ethyl acetate and the solvent was evaporated to remove the solvent under reduced pressure, after which the product obtained was recrystallized from ethanol to obtain 0.67 g of the compound mentioned in the preamble is colorless crystals.

Example CCVIII

N- [2- (4-Chloro-N-methylcinnamylamino) ethyl] -N- (2-piperidinoethyl) -5-isoquinolinesulfonamide.

To a solution of 0.39 g of amorphous compound obtained in Example CXC, 0.145 g of 1-piperidinethanol and 0.399 g of triphenylphosphine in 5 ml of tetrahydrofuran was added dropwise a solution of 0.245 g of diethyl azodicarboxylate in 2 ml of tetrahydrofuran with stirring and cooling with ice. The mixture was stirred and evaporated for 2 hours to remove the solvent under reduced pressure and the resulting residue was dissolved in 30 ml of ethyl acetate and extracted three times with 10 ml of 1N hydrochloric acid. The aqueous layer was made alkaline with sodium bicarbonate and extracted three times with 10 ml of ethyl acetate and the organic extract was dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting residue was applied to a silica gel column and eluted with 2% methanol in chloroform to obtain 0.37 g of the title compound 40 as a colorless oil.

To the solution of the above oil in 3 ml of methanol was added 0.5 ml of 4N hydrochloric acid in ethyl acetate and the whole was evaporated to remove the solvent under reduced pressure. Ether was added to the concentrate to form a powder which was then collected by filtration to obtain 0.35 g of the corresponding trihydrochloride in the form of a colorless powder.

45

Example CCIX

N- [2- (4-Chloro-N-methylcinnamylamino) ethyl] -N- (2-dimethylaminoethyl) -5-isoquinolinesulfonamide.

To a solution of 1.0 g of amorphous compound obtained in Example CXC, 0.267 g of 2-dimethylaminoethanol and 0.992 g of triphenylphosphine in 5 ml of tetrahydrofuran was added dropwise a solution of 0.652 g of diethyl azodicarboxylate in 2 ml of tetrahydrofuran with stirring and cooling with ice.

After 2 hours, the reaction mixture was evaporated under reduced pressure to remove tetrahydrofuran and the resulting residue was dissolved in 10 ml of ethyl acetate and extracted three times with 10 ml of 1N hydrochloric acid. The aqueous layer was made alkaline with sodium bicarbonate and extracted three times with 10 ml of ethyl acetate and the organic layer was dried over magnesium sulfate and evaporated to remove the solvent under reduced pressure. The resulting residue was then applied to a silica gel column and eluted with 3% methanol in chloroform to give 0.77 g of the title compound as a colorless oil.

49 194549

To a solution of the oil thus obtained in 5 ml of methanol was added 1.4 ml of 4N hydrochloric acid in ethyl acetate and after evaporation of the solvent under reduced pressure, ether was added to the resulting concentrate to form a powder which was then collected by filtration and dried to obtain 0.7 g of the corresponding trihydrochloride in the form of a powder.

5

Example CCX

N- (2-Piperidinoethyl) -N- [2- (N-methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.

The amorphous product obtained in example CLXXIII was treated in accordance with the method described in example CXC to obtain N- [2- (N-methylcinnamylamino) ethyl] -5-10 isoquinolinesulfonamide.

To a solution of 0.476 g of the above compound, 0.193 g of 1-piperidinethanol and 0.524 g of triphenylphosphine in 5 ml of tetrahydrofuran was added a solution of 0.348 g of diethylazodicarboxylate in 2 ml of tetrahydrofuran with stirring and cooling with ice and the mixture was stored for 3 hours and evaporated to remove the solvent under reduced pressure. 30 ml of ethyl acetate was added to the concentrate and the mixture was extracted three times with 10 ml of hydrochloric acid. The extract was made alkaline with sodium bicarbonate and extracted three times with 10 ml of ethyl acetate. The ethyl acetate-containing solution was dried over magnesium sulfate and evaporated under reduced pressure to remove the solvent. The resulting residue was applied to a silica gel column and eluted with 5% methanol in chloroform to obtain 0.44 g of the title compound as a colorless oil.

To the oil thus obtained in 5 ml of methanol was added 0.8 ml of 4N hydrochloric acid in ethyl acetate and the solution was evaporated to remove the solvent under reduced pressure. A precipitate obtained by adding 50 ml of ether was collected by filtration and dried to give 0.4 g of the corresponding trihydrochloride as a colorless powder.

25

Example CCXI

N-Anisyl-N- [2- (4-chloro-cinnamylamino) -ethyl] -5-isoquinolinesulfonamide.

In 10 ml of tetrahydrofuran, 0.4 g of crystals obtained in Example CLXXII, 0.276 g of anisyl alcohol and 0.524 g of triphenylphosphine were dissolved, and a solution of 0.404 g of diisopropylazodicarboxylate in 2 ml of tetrahydrofuran was added dropwise with stirring and cooling with ice. The reaction mixture was warmed to room temperature and was stored overnight and then evaporated under reduced pressure to remove the solvent. The resulting residue was dissolved in 30 ml of ethyl acetate and the mixture was extracted twice with 30 ml of 1N hydrochloric acid. The extract was made alkaline with sodium bicarbonate and extracted twice with 30 ml of ethyl acetate. The ethyl acetate solution was washed with water, dried over magnesium sulfate and evaporated under reduced pressure to remove the solvent. The resulting residue was applied to a silica gel column and eluted with 1% methanol in chloroform to give 0.22 g of the title compound as a colorless oil.

Example CCX »N - [2- (4-Chloro-cinnamylamino) -ethyl] -N-phenethyl-5-isoquinolinesulfonamide.

The process described in Example CCXI was repeated except that 0.146 g of phenethyl alcohol was used instead of anisyl alcohol to obtain 0.37 g of the title compound as a colorless oil.

45

Example CCXIII

N-Benzyl-N- [2- (4-chloro-cinnamylamino) -ethyl] -5-isoquinolinesulfonamide.

The process described in Example CCXI was repeated except that 0.162 g of benzyl alcohol was used instead of anisyl alcohol to give 0.3 g of the title compound as a colorless oil.

Example CCXIV

N- [2- (4-Chlorocinnamylamino) ethyl] -N-methyl-5-isoquinolinesulfonamide.

The process described in Example CCXI was repeated except that 48 mg of methanol was used instead of anisyl alcohol to obtain 0.3 g of the title compound as a colorless oil.

194549 50

Reference Example 45 N- (3,4-Dimethoxyphenyl) -5-isoquinolinesulfonamide.

3.06 g of 3,4-dimethoxyaniline was dissolved in 30 ml of pyridine, 5.28 g of 5-isoquinolinesulfonyl chloride.HCl was added in small amounts with stirring and cooling with ice and the mixture was stirred for 30 minutes and further stirred at room temperature overnight. After evaporating the pyridine under reduced pressure and adding 20 ml of water, the mixture was extracted twice with 50 ml of chloroform / isopropanol (10: 1). The extract was dried over magnesium sulfate and evaporated under reduced pressure to remove the solvent. To the resulting residue, 20 ml of benzene / chloroform (3: 1) was added and the mixture was slightly heated and collected to obtain 5.64 g of the title compound as colorless crystals.

Reference Example 46 N- (3,4-Dimethoxyphenyl) -N- (2-phthalimide-ethyl) -5-isoquinolinesulfonamide.

500 mg of crystals obtained in Reference Example 45 were dissolved in 7 ml of dimethylformamide and 4 ml of tetrahydrofuran, 70 mg of 60% sodium hydride was added to the solution with stirring and cooling with ice and the mixture was stirred for 20 minutes and after addition of 406 mg of bromomethyl phthalimide, the mixture was refluxed for 6 hours with stirring. After the addition of 10 ml of ice water, the reaction mixture was extracted with 30 ml of ethyl acetate and the extract was dried over magnesium sulfate and evaporated under reduced pressure to remove the solvent. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (100: 1) to obtain 320 mg of the title compound as colorless crystals.

Reference Example 47 N- (3,4-Dimethoxyphenyl) * N- (2-aminoethyl) -5-isoquinolinesulfonamide.

517 g of crystals obtained in Reference Example 46 were dissolved in 5 ml of methanol and 5 ml of chloroform, 60 mg of hydrazine hydrate was added to the solution and the mixture was refluxed for 3 hours.

Crystallized, insoluble material was filtered off and the filtrate was evaporated under reduced pressure to remove the solvent. After adding 10 ml of ethyl acetate, the mixture was filtered to remove the insoluble material and then evaporated under reduced pressure to obtain 420 mg of the title compound obtained as a light yellow oil.

Example CCXV

N- (3,4-Dimethoxyphenyl) -N- [2- (4-chloro-cinnamylamino) -ethyl] -5-isoquinolinesulfonamide.

320 mg of oil obtained in Reference Example 47 were dissolved in 6 ml of dimethylformamide, 200 mg of potassium carbonate and 150 mg of p-chloro-cinnamyl chloride were added to the solution and the mixture was stirred at room temperature overnight. After adding 20 ml of water, the reaction mixture was extracted twice with 30 ml of chloroform. The extract was washed with an aqueous, saturated solution of sodium chloride, dried over magnesium chloride and evaporated under reduced pressure to remove the solvent. The resulting residue was applied to a silica gel column and eluted with chloroform / methanol (100: 1) to obtain 90 mg of the title compound as colorless crystals.

45 Example CCXVI

N- {2- [Bis (4-chloro-cinnamyl) amino] ethyl} -5-isoquinolinesulfonamide.

In Example CCXV, for eluting using chloroform / methanol, eluting was carried out using chloroform to obtain 100 mg of the title compound in a colorless, amorphous form.

As described above, the following compounds were prepared.

Example CCXVII

N- [2- (4 * Methoxy-α-methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.2 HCl.

Colorless, amorphous form.

51 194549

Example CCXVIII

N- [2- (4-Methoxycarbonyl-N, α-dimethylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.2 HCl.

Colorless, amorphous form.

Example CCXIX

N- [2- (4-Methoxy-N, α-dimethylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Colorless oil.

Example CCXX

W N- (2-Methylaminoethyl) -N- [2- (4-chloro-N-methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Colorless oil.

Example CCXXI

N- (2-Methylaminoethyl) -N- [2- (4-chloro-N-methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.3 HCl.

15 Colorless, amorphous form.

Example CCXXII

N- (2-Hydroxyethyl) -N- [2- (4-methoxy-N, α-dimethylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Colorless, amorphous form.

20

Example CCXXIII

N- [2- (Methoxy) ethyl] -N- [2- (N-methyl-4-methoxy-α-methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide. Colorless oil.

Example CCXXIV

N- [2- (4-Chloro-cinnamylamino) -ethyl] -N- (2-dimethylaminoethyl-5-isoquinoline sulfonarnide.3 HCl.

Colorless, amorphous form.

Example CCXXV

30 N [2- (4-Chlorocinnamylamino) ethyl] -N- (2-methylaminoethyl) -5-isoquinolinesulfonamide.

Colorless, amorphous form.

Example CCXXVI

N-2- (4-Chlorocinnamylamino) ethyl] -N- (2-pyridylmethyl) -5-isoquinolinesulfonamide.3 HCl.

35 Light yellow, amorphous form.

Example CCXXVH

N- [2- (4-Chlorocinnamylamino) ethyl] -N- (3-pyridylmethyl) -5-isoquinolinesulfonamide.3 HCl.

Light yellow, amorphous form.

40

Example CCXXVIII

N- [2- (3,4-dimethoxy-α-methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.2 HCl.

Yellow, amorphous form.

45 Example CCXXIX

N- [2- (α-Methyl-3,4,5-trimethoxycinnamylamino) ethyl] -5-isoquinolinesulfonamide.2 HCl.

Example CCXXX

N- (2-Dimethylaminoethyl) -N- [2- (N-methyl-3,4,5-trimethoxycinnamylamino) ethyl] -5-isoquinolinesulfonamide.

50 Colorless oil.

Example CCXXXI

N- (2-Dimethylaminoethyl) -N- [2- (N-methyl-3,4,5-trimethoxycinnamylamino) ethyl] -5-isoquinolinesulfonamide.3 HCl.

55 Yellow, amorphous form.

194549 52

Example CCXXXII

N- [2- (4-Chloro-cinnamylamino) -ethyl] -N- (3,4,5-trimethoxy-benzyl) -5-isoquinolinesulfonamide-2 HCl.

Colorless, amorphous form.

Example CCXXXIII

N-Cyanomethyl-N- [2- (4-methoxycarbonyl-N-α-dimethylcinnamylarnino) ethyl] -5-isoquinoline sulfonamide. Colorless oil.

Example CCXXXIV

N-Cyanomethyl-N- (2- (4-methoxycarbonyl-N, α-dimethylcinnamylamino) ethyl] -5-isoquinolinesulfonamide-2 HCl Colorless, amorphous form.

Example CCXXXV

N- (2-Dimethylaminoethyl) -N- [2- (4-methoxycarbonyl-N, α-dimethylcinnamylamino) ethyl] -5-15 isoquinolinesulfonamide.

Colorless oil.

Example CCXXXVI

N- (2-Dimethylaminoethyl) -N- [2- (4-methoxycarbonyl-N, α-dimethylcinnamylamino) ethyl] -5-20 isoquinolinesulfonamide.3 HCl.

Colorless, amorphous.

Example CCXXXVII

N- (2-Morpholinoethyl) -N- [2- (4-methoxycarbonyl-N, α-dimethylcinnamylamino) ethyl] -5-isoquinolinesulfonamide. 25 Colorless oil.

Example CCXXXVIII

N- (2-Morpholinoethyl) -N- [2- (4-methoxycarbonyl-N, α-dimethylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.3 HCl.

30 Colorless, amorphous form.

Example CCXXXIX

N- (2-Aminoethyl) -N- [2- (4-methoxycarbonyl-N, α-dimethylcinnamylamino) ethyl] -5-isoquinoline sulfonamide. Colorless oil.

35

Example CCXL

N- (2-Aminoethyl) -N- [2- (4-methoxycarbonyl-N, α-dimethylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.3 HCl.

Colorless, amorphous form.

40

Example CCXLI

N- [2- (4-Chloro-N-methylcinnamylamino) ethyl] -N-methoxycarbonylmethyl-5-isoquinolinesulfonamide.2 HCl. Yellow, amorphous form.

45 Example CCXLII

N-Carboxymethyl-N- [2- (4-chloro-N-methylcinnamylamino) ethyl] 5-isoquinolinesulfonamide.

Light brown, amorphous form.

Example CCXLIII

50 N- [2-Carboxymethyl-4-chloro-cinnamylamino) -ethyl] -N-methyl-5-isoquinolinesulfonamide.

Light yellow, amorphous form.

Example CCXLIV

N- [2- (4-Chloro-N-methylcinnamylamino) ethyl] -N-methyl-5-isoquinolinesulfonamide.2 HCl.

55 Light brown, amorphous form.

53 194549

Example CCXLV

N-Carbamoyl-N- [2- (4-chloro-N-methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.2 HCl.

Colorless, amorphous.

Example CCXLVI

N- [2- (4-Chlorocinnamylamino) ethyl] -N - [(5-methyl-4-imidazolyl) methyl] -5-isoquinolinesulfonamide.HCl. Light yellow, amorphous form.

Example CCXLVII

10 N- [2- (4-Chloro-N-methoxycarbonylmethylcinnamylamino) ethyl] -N-methyl-5-isoquinolinesulfonamide.2 HCl. Yellow, amorphous form.

Example CCXLVIII

N- [2- (N-Carbamoylmethyl-4-chloro-cinnamylamino) -ethyl] -N-methyl-5-isoquinolinesulfonamide-2 HCl.

15 Light yellow, amorphous form.

Example CCIL

N- [2- (4-Chloro-N-cyanomethylcinnamylamino) ethyl] -N-methyl-5-isoquinolinesulfonamide.2 HCl.

Light brown, amorphous form.

20

Example CCL

N- [2- (4-Chloro-N-methylcinnamylamino) ethyl] -N-morpholinocarbonylmethyl-5-isoquinolinesulfonarnide.

Colorless oil.

Example CCLI

N- {2- [N- (2-Aminoethyl) -4-chloro-cinnamylamino] ethyl} -N-methyl5-isoquinolinesulfonamide.3 HCl.

Colorless, amorphous form.

Example CCLII

N- [2- (4-Chloro-N-methylcinnamylamino) ethyl] -N- [2- (1-piperazinyl) ethyl] -5-isoquinolinesulfonamide.4 HCl. Colorless, amorphous form.

Example CCLIII

N- [2- (4-Chloro-N-methylcinnamylamino) ethyl] -N- [2- (4-methyl-1-piperazinyl) ethyl-5-isoquinolinesulfonamide.4 HCl.

Colorless, amorphous form.

Example CCLIV

N- [2- (3-Methoxy-α-methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.2 HCl.

40 Light yellow crystals.

Example CCLV

N- [2- (4-Hydroxymethyl-α-methyl-cinnamylamino) -ethyl] -5-isoquinolinesulfonamide.

Colorless crystals.

45

Example CCLVI

N- [2- (α-Methyl-4-methylthiocinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Colorless, amorphous form.

50 Example CCLVII

N- [2- (α-Methyl-4-methylsulfonylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Colorless, amorphous form.

Example CCLVIII

55 N- [2- (α-Methyl-4-methylsulfonyl cinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Colorless, amorphous.

«194549 54

Example CCLIX

N- [2- (4-Cyano-α-methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Colorless needles.

Example CCLX

N- [2- (4-Carbamoyl-α-methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Colorless needles.

Example CCLXI

N- [2- (4-acetamide-α-methyl-cinnamylamino) -ethyl] -5-isoquinoline sulfonamide.

Colorless, amorphous form.

Example CCLXII

N- [2- (3-Nitro-3-methoxy-α-methylcinnamylamino) ethyl] -5-isoquinoline sulfonamide.HCl.

15 Colorless crystals.

Example CCLX III

N- [2- (2-Methoxy-α-methylcinnamylamino) ethyl] -5-isoquinolinesulfonamide.

Colorless, amorphous form.

20

Example CCLXtV

The following experiments were conducted to confirm the use of the above compounds of the invention.

25 Relaxation effect on the smooth muscle of blood vessels (V.R. EDS0).

A rabbit was killed by bleeding and the upper portion of the mesenteric vein was removed and cut into a spiral shape to prepare a band-shaped sample by a conventional method. The sample was suspended by applying a voltage to a Krebs-Henseleit solution, bubbling an oxygen gas containing 5% carbon dioxide. The sample was contracted by adding potassium chloride to maintain a predetermined voltage. A test compound was then administered cumulatively. The relaxation activity of the test compound is expressed by ED (pM), i.e., a concentration of the compound that relaxes the stress to 50% of the stress obtained in the presence of potassium chloride (as 100%) only.

The inhibition of platelet aggregation (P.A .; IC &).

(1) The preparation of washed plates.

The blood was obtained from a healthy person and mixed with one tenth volume of 0.38% sodium citrate and the mixture was centrifuged at 700 x g for 10 minutes to obtain a platelet-rich plasma (PRP). To the PRP, one sixth volume of the 40 ACD solution (2.2% sodium citrate, 0.8% citric acid and 2.2% glucose, freshly prepared for use) was added and the mixture was centrifuged at 1500 xg for 10 minutes to obtaining a layer of plates.

Subsequently, the platelet layer was suspended in a modified HEPES-Tyrode solution (135 mM NaCl, 2.7 mM KCl, 1 mM MgCl 2, 0.1 mg / ml glucose, 20 mM HEPES, pH = 7.4). One sixth volume of ACD solution was added to this suspension and the whole was further centrifuged at 1500 x g 45 for 5 minutes to prepare a platelet layer. The platelet layer was then suspended in a modified HEPES Tyrode solution to obtain approximately 3 x 105 / μΙ in washed platelet suspension.

(2) Measurement of the caking of the plates.

To 270 µl of washed platelet suspension was added 3 μΙ of a solution of a 50 test compound, dissolved in a suitable mixture in various concentrations and the mixture was stored at 37 ° C for 2 minutes. After adding 30 μΙ of a 20 pg / ml solution of collagen, the absorbance was measured with a 4-channel analyzer to determine the agglomeration (HEMA Tracer 601; Niko Bioscience).

(3) Determination of the influence on the compounds to be investigated.

55 For comparison, the above method was carried out, except that the mixture to be determined was used without the test compound, and the absorbance before the addition of collagen and the maximum absorbance after addition of collagen were measured and the difference between the two absorbances was 55 194549 taken as 100% caking or agglutinating.

For the test compound, the absorbance before collagen addition and the maximum absorbance after collagen addition were measured and 1% inhibition was determined compared to the comparative test. A concentration of the test compound giving 50% inhibition was expressed as IC

Calmodulin-dependent phosphodiesterase inhibition.

(1) The preparation of calmodulin-dependent phosphodiesterase (Ca 2+ PDE).

Ca 2+ PDE was partially purified from the brain of a rat by DEAE-Sepharose column chromatography. (2) Preparation of calmodulin.

Calmodulin was purified from the brains of a calf using a column with an affinity for the inhibitory W-7 agent for calmodulin.

(3) Measurement of Ca 2+ PDE activity.

A reaction mixture containing 20 μl 500 mM Tris-HCl (pH = 8.0), 20 μΙ 50 mM MgCl 2, 20 μΙ 2 mM CaCl 2 (or 15 10 mM EGTA), 20 μ11 mg / ml bovine serum albumin, PDE, 200 mg calmodulin, sample to be tested and distilled water to bring the total volume to 200 μΙ. To the mixture was added 20 μΙ of 4 μΜ [3 H] -cGMP (2.5 Ci / ml), after which the mixture was stored at 30 ° C for 15 minutes and then heated in boiling water for 3 to 5 minutes to complete the reaction. and cooled in an ice-water bath. 20 μο 5'-nucleotidase (Snake venum) was added to the mixture and the mixture was again stored at 30 ° C for 10 minutes. After addition of about 2 ml of water, the sample was applied to a column of a cation exchanger (Biorad AG.AG50W-X4 to adsorb the [3 H] -guanosine and further about 2 ml of washing water before the sample was added to the column. was washed with about 20 ml of water and the adsorbed [3 H] -guanosine was eluted with 3 ml of 3N NH 4 OH and the eluate was immediately collected in an ampoule After adding 10 ml of an emulsified solution of a signal-releasing agent (ACS) -II, AMERSHAM) the radioactivity was measured using a LS7500 scintillation counter (Beckmann), the enzyme activity in the presence of calmodulin was set at 0% and the concentration of the test compound in μΜ giving 50% inhibition was expressed as ΙΟ ^.

The results are shown in the following table.

Example V.R. (ED *,) P.A. (IC, *,) Ca 2+ PDE (IC, *,) (μΜ) (μΜ)

35 I

II 47

III

IV 12 53

V-VI

40 VII 1.6 13 VIII 1.8 51

IX

X 6.4 37 XI 59

45 XII

XIII 1.2

XIV

XV 43

XVI - XVIII

50 XIX 22 20 XX 1 23

XXI

XXII 1.8 XXIII 0.25 66 55 XXIV 8.3 63 XXV 0.55 70 194549 56 example V.R. (ED50) P.A. (IC, *) Ca 2+ PDE (IC. *) _ (ΜΜ) (μΜ) O _________________ XXVI 10

XXVII-XXVIII

XXIX 7.6 73

XXX

10 XXXI 24 ± 7.0

XXXII

XXXIII 21 XXXIV 1.8 XXXV 6.9 75 XXXVI 0.81 10.5 XXXVII 4.4 29 XXXVIII 8.1

XXXIX

on XL 4.7 20 U XLI 1.8 10 XLII 6.2 XLIII 0.36 3.8

XLIV

XLV 7.5 XLVI 4 36 XLVII 0.39 18 XLVIII 3.9 6.5 IL 0.92 L 0.67 70 11 30 LI 0.17 38

Lil 9.9

L111 LIV 1.7 35 LV 8'6 LVI 1.2 39 LVII 0.75 50 LVIII 0.25 63

LIX

LX 10 40 LXI 10 LXII 2.3

LXIII-LXVI

LXVII 2.2 3.6 LXVIII 3.3 ± 1.4 6 45 LXIX 14

LXX

LXXI 14 93 LXXII 4 6.2 LXXIII 14 21 50 LXXIV 2.8 LXXV 9.7 LXXVI 2.9 LXX VII 1.4 LXXVIII 1.8 55 LXX IX 5.5 57 194549 example V.R. (ED ^,) P.A. (IC. *) Ca2 + PDE (IC *,) 5 (μΜ) (μΜ) LXXX 2.4 13 LXXXI 1.7 32 LXXXII 0.86 33 LXXXIII 0.39 24 10 LXXXIV 43 LXXXV 75 LXXXVI 13 24 LXXXVII 1, 7 1.9 LXXXVI II 4.3 14

15 LXXXIX

XC 0.33 3.4 XCI 1.1 XCII 63 XCIII 0.31 3.4

XCIV-XCVI

XCVII 39 XCVIII 22 IC 2.8 61 25 C 95

Cl 29

Cll 24

CIII-CIV

CV 5.4 12 CVI 8.9 28

30 CVII

CVIII 1.2 10 CIX 0.59 1.3 CX 3.8 9.3 CXI 7.0 77

35 CXII

CXIII 0.88 20 1.2 CXIV 54 100

CXV

CXVI 1.5

CXVII-CXVIII

CXIX 11 51 CXX 19 ± 3.0

CXXI-CXXII

CXXIII 4.8 22 45 CXXIV 11 CXXV 9.2 CXXVI 10 CXX VII 39 CXXVIII 1.8 50 CXXIX 0.82 CXXX 13 CXXXI 1.5

CXXXII-CXLI

CXLII 6.8 1.1. CXLIII 0.82 1.5 194549 58 Example V.R. (ED ^) P.A. (IC, *,) Ca2 + PDE (IC50) 5 (μΜ) (μΜ) CXLIV 1.8 1.0 CXLV 12 CXLVI 2.8 CXLVII 1.2 42

U CXLVIII-CLII

CLIII 1.5 25 CLIV 60 CLV 4.1 55 CLVI 4.4 19 19 ® CLVII 3.2 36 8.6 CLVIII 17 21 CLIX 5.1 24 92 CLX 1.8 56 20 CLXI 4 · 7 CLXII 35 CLXI II 31 CLXIV 4.1 CLXV 11 CLXVI 1.4 90 * CLXVII 1.2 52 CLXVIII 3.6 26 CLXIX 4.6 CLXX 5.2 CLXXI 5.4 CLXXII 1.0 51 9.4

CLXXIII

CLXXIV 11 CLXXV 2.0 13 CLXXVI 1.2 55 8.5 CLXXVII 2.7 56 24 CLXXVIII 2.4 50 CLXXIX 2.8 51 7.8 CLXXX 1.8 4.8 CLXXI 8.4 38 4.7 40 CLXXII 17 CLXXIII 2.8 CLXXIV 2.8 12 CLXXV 0.21 „CLXXVI-

CLXXVII

CLXXXVIII 4 189-1 0.22 2.7 189-11 0.20 189-111 0.29 1.0 CXC 0.19 CXCI 1.9 26 CXCII 0.12 84 CXCI II 3.2 12 CXCIV 80

5 CXCV-CC

Claims (2)

59 194549 example V.R. (ED *,) P.A. (IC. *,) Ca2 + PDE (IC, *) 5 (μΜ) (μΜ) CCI 3.2 CCII 1.3 CCIII-CCXVI It appears that certain compounds according to the present invention have an inhibitory effect on the platelet clotting, as well as inhibitory action against protein kinase A, light chain myosin kinase, protein kinase C, calmodulin dependent protein kinase 11, cyclic AMP dependent phosphodiesterase and the like, but have little influence on cardiac functions. As is apparent from the above results, the present compounds have a smooth muscle relaxation activity as mentioned above and are therefore suitable as a vasodilatory agent or as a means of improving circulation in the brain; and because the present compound has an inhibitory effect on platelet aggregation, they are also suitable as a prophylactic or therapeutic agent for thrombosis. In addition, because the compounds have an inhibitory action against various kinases, they are suitable as anti-tumor agents. The above compounds have low toxicity and are therefore useful as pharmaceutical preparations. Thus, the invention relates to novel quinoline sulfonamod derivatives with a relaxation activity with regard to the smooth muscle of blood vessels as well as an inhibitory effect with respect to platelet aggregation and an inhibitory effect with regard to protein kinase A, myosin-light chain kinase, protein kinase C and calmodulin-dependent proteinase 11, but has little influence on the function of the heart. Conclusions Isoquinoline sulfonamides with a relaxation activity for the smooth muscles of blood vessels and a low toxicity, characterized in that these compounds correspond to formula 1 on the formula sheet, wherein Y represents N or H 3 C-N and R 1 represents a hydrogen atom, an optionally substituted lower alkyl group, a formyl group, a halophenylpropargyl group, an optionally substituted aralkyl group or optionally substituted phenyl group, and R 2 represents a group represented by formula 3 on the formula sheet, wherein R 10 represents a hydrogen atom, a nitro group, a optionally substituted amino group, an optionally substituted hydroxyl group, a lower alkyl group or a halogen atom, or R1 and R10 together represent a lower alkylene group, a hydrogen atom, a hydroxyl group or a lower alkoxy group, R12 and R13 each represent a hydrogen atom or together = 0 Ar represents a phenyl group, a naphthyl group or a heterocyclic group, 45. is an integer from 1 to 3 and a group> CR14 R15 or> NR14, where R14 is a hydrogen atom, an optionally substituted hydroxyl group, an optionally substituted phenyl group , an acyl group, a substituted c arbonyl group, an optionally substituted alkoxycarbonyl group, a substituted carbamoy group, an optionally substituted amino group, an arylsulfonyl group, an aralkylsulfonyl group, an aralkyl group 50 or a heterocyclyl group, and R15 represents a hydrogen atom or a lower alkyl group or R15 and R14 together represent an alkylene dixy 0, 194549 60 and quaternary ammonium salts form salts of the compound of formula 1 and non-toxic salts of the compound of formula 1. A pharmaceutical composition containing a compound according to claim 1. Hereby 4 sheets of drawing