HK1010871A - New bisimide compounds, process for preparing them and pharmaceutical compositions containing them - Google Patents

Description

New bisimide compounds, process for their preparation and pharmaceutical compositions containing them

The present invention relates to novel bisimide compounds, to a process for their preparation, to pharmaceutical compositions containing them and to their use in the preparation of pharmaceutical compositions for the treatment of cancer.

Compounds derived from bisimides have been described in the literature. In particular those described in patents EP 506008, DE 4034687, WO 9500490 or DE 4232739.

The compounds of the present invention have a completely novel structure compared to the compounds described in the prior art. They are symmetrical or asymmetrical compounds which have not been described before. Furthermore, due to their strong pharmacological activity, they are particularly suitable as new drugs for the treatment of cancer, in particular solid tumors.

More particularly, the invention relates to compounds of formula (I)

Wherein:

m and n, which may be the same or different, represent 0 or 1,

z and Y, which may be the same or different, represent hydrogen, a halogen atom or a linear or branched (C)₁-C₆) Alkyl, straight-chain or branched (C)₁-C₆) Trihaloalkyl, straight or branched (C)₁-C₆) Alkoxy, hydroxy, cyano, nitro, amino, alkylamino or dialkylamino,

z represents a linear or branched C₄To C₁₂Alkylene chain, one or more-CH therein₂The group being selected by any one of the following atoms or groupsSelective substitution: -NR- (wherein R represents a hydrogen atom or a linear or branched (C)₁-C₆) Alkyl), -0-, -S-, -SO-, -SO-₂-or-CONH-, or a substituted or unsubstituted heterocyclic group,

a is formed with two adjacent carbon atoms on the phenyl ring:

-a substituted or unsubstituted benzene ring,

-a substituted or unsubstituted naphthalene ring,

-a substituted or unsubstituted tetrahydronaphthalene ring or a substituted or unsubstituted 1, 4-dioxo-1, 2, 3, 4-tetrahydronaphthalene ring,

-or a substituted or unsubstituted heterocyclic ring,represents any one of the following groups:

wherein:

m₁and n₁Which may be identical or different, represent 0 or 1,

Z₁and Y₁Which may be the same or different, represent hydrogen, a halogen atom or a linear or branched (C)₁-C₆) Alkyl, aryl, heteroaryl, and heteroaryl,

Straight-chain or branched (C)₁-C₆) Trihaloalkyl, straight or branched (C)₁-C₆) Alkoxy radical

A group, a hydroxyl group, a cyano group, a nitro group, an amino group, an alkylamino group or a dialkylamino group,

A₁formed with two adjacent carbon atoms on the phenyl ring:

-a substituted or unsubstituted benzene ring,

-a substituted or unsubstituted naphthalene ring,

a substituted or unsubstituted tetralin ring or a substituted or unsubstituted 1, 4-dioxy-

1, 2, 3, 4-tetrahydronaphthalene ring,

-a substituted or unsubstituted heterocyclic ring,

wherein Z₂And Y₂May be the same or different and represents hydrogen, a halogen atom or a linear or branched (C)₁-C₆) Alkyl, straight-chain or branched (C)₁-C₆) Trihaloalkyl, straight or branched (C)₁-C₆) Alkoxy, hydroxy, cyano, nitro, amino, alkylamino or dialkylamino, isomers thereof and addition salts thereof with a pharmaceutically acceptable acid or base.

Pharmaceutically acceptable acids which may be mentioned include (without any limiting meaning): hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, acetic acid, trifluoroacetic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, tartaric acid, maleic acid, citric acid, ascorbic acid, oxalic acid, methanesulfonic acid, camphoric acid, and the like.

Pharmaceutically acceptable bases which may be mentioned include (without any limiting meaning): sodium hydroxide, potassium hydroxide, triethylamine, tetrabutyl amine, and the like.

The terms substituted or unsubstituted phenyl, substituted or unsubstituted naphthyl and substituted or unsubstituted tetrahydronaphthyl are understood to mean (C) which is linear or branched, substituted or unsubstituted by one or more halogen atoms₁-C₆) Alkyl, straight-chain or branched (C)₁-C₆) Trihaloalkyl, straight or branched (C)₁-C₆) Alkoxy, hydroxy, nitro or cyano or amino (substituted by one or more straight or branched chains of (C)₁-C₆) Alkyl optionally substituted) optionally substituted phenyl, naphthyl or tetrahydronaphthyl.

The term substituted or unsubstituted heteroRing is understood to mean a saturated or unsaturated 5-to 16-membered monocyclic or bicyclic ring containing 1, 2 or 3 heteroatoms selected from oxygen, nitrogen and sulfur, it being understood that the heterocyclic ring may be interrupted by one or more halogen atoms or be straight-chain or branched (C)₁-C₆) Alkyl, straight-chain or branched (C)₁-C₆) Alkoxy, hydroxy, trihalomethyl, nitro or cyano radicals or amino radicals, substituted by one or more straight or branched (C)₁-C₆) Alkyl optionally substituted) optionally substituted.

Preferred compounds according to the invention are those in which:

a is formed with two adjacent carbon atoms on the phenyl ring:

-a substituted or unsubstituted naphthalene ring,

-or a substituted or unsubstituted heterocycle, preferably selected from the group consisting of substituted or unsubstituted indole,

Substituted or unsubstituted benzo [ b ] thiophene and substituted or unsubstituted benzo [ b ] furan rings.

Preferred components are those in which Z represents C₄To C₁₂Alkylene chain, 1, 2 or 3-CH in the chain₂The radical being substituted by 1, 2 or 3-NR-radicals (in which R represents a hydrogen atom or a linear or branched (C)₁-C₆) Alkyl) is substituted.

The invention also includes a process for the preparation of compounds of formula (I) using as starting material an anhydride of formula (II):wherein A, X, Y, m and n are as defined for formula (I), with an excess of an amine of formula (III):

H₂N-Z-NH₂(III) wherein Z is as defined for formula (I) and is isolated to yield: -a compound of formula (I/a), a particular case of compounds of formula (I):wherein A, Z, Y, m and n have the same meanings as in formula (I), -or (IV):wherein A, Z, Y, m and n have the same meanings as in formula (I), -subjecting it to the following reaction:

reacting with an anhydride of formula (II/a):wherein A is₁，Z₁，Y₁，m₁And n₁As defined for formula (I) to give a compound of formula (I/b) -a particular case of compounds of formula (I):wherein A, Z, Y, m, n, Z, A₁，Z₁，Y₁，m₁And n₁Has the same meaning as in formula (I),

or with an anhydride of formula (V):wherein Z₂And Y₂As defined for formula (I) to give a compound of formula (I/c) -a particular case of compounds of formula (I):wherein A, Z, Y, Z₂，Y₂And Z, m and n have the same meanings as in formula (I), and if desired, the compounds of formula (I/a), (I/b) or (I/c) can be subjected to the standard reaction for converting the substituents on the aromatic ring and, where appropriate, purified according to standard purification techniques, the isomers of which can be selectively separated according to standard separation techniques and, if desired, converted into addition salts thereof with a pharmaceutically acceptable acid or base.

The anhydrides of the formula (II) or (V) are commercially available or can be prepared according to known methods.

The invention also encompasses pharmaceutical compositions comprising as active ingredient at least one compound of formula (I) and one or more inert, non-toxic suitable excipients. More specifically, the pharmaceutical compositions of the present invention are in a form suitable for oral, parenteral (intravenous or subcutaneous) or intranasal administration, such as plain or sugar-coated tablets, sublingual tablets, gelatin capsules, lozenges, suppositories, creams, ointments, dermal gels, injectable preparations, drinkable suspensions and the like.

The appropriate dosage can be taken according to the nature and severity of the disease, the route of administration, and the age and weight of the patient. The dosage ranges from 0.1 to 400 mg/day, and can be administered in one or more divided doses.

The following examples are illustrative of the present invention, but are not intended to be limiting in any manner.

The starting materials used are known products or prepared according to known methods.

The following preparation methods may yield synthetic intermediates useful in the preparation of the compounds of the present invention.

The structures of the compounds described in the examples and preparations were determined according to conventional spectroscopic techniques (infrared, NMR, mass spectrometry, etc.). Preparation A: 10-methoxy-2-oxacyclopenta [ c ] phenanthrene-1, 3-dione step a: 2-hydroxymethylene-7-methoxy-3, 4-dihydro-2H-naphthalen-1-one

The desired product was prepared from 7-methoxy-1-tetralone and ethyl formate following the procedure described in organic synthesis 1959, 39, 27. And B: 3- (7-methoxy-1-oxo-3, 4-hydro-1H-naphthalen-2-ylidene) -propionic acid tert-butyl ester

A solution of 5mmol of the product prepared in the previous step and 6mmol of tert-butyl triphenylphosphoranylidene acetate in 90ml of dichloromethane is stirred at room temperature under argon for 2 hours. The solvent was concentrated under pressure and the residue was crystallized from isopropyl ether. The precipitate was filtered off and the filtrate was concentrated to give 95% yellow crystalline product. Melting point: 76 ℃ step C: 9-methoxy-5, 6-dihydrobenzo [ h ] -benzopyran-2-one

To a solution of 4.6mmol of the product prepared in the previous step in 10ml of dichloromethane was added dropwise 6.5ml of trifluoroacetic anhydride and 3.6ml of trifluoroacetic acid in that order at room temperature. The solution was stirred for 4 hours and then hydrolyzed with 30ml of water. The aqueous phase is extracted with 30ml of dichloromethane and the organic phase is washed with water, dried and concentrated in vacuo. Melting point: 108-: 6-methoxy-9, 10-dihydrophenanthrene-3, 4-dicarboxylic acid dimethyl ester

A solution of 5.3mmol of the product prepared in the previous step and 2.6ml of methyl acetylenedicarboxylate in 20ml of dimethylformamide was refluxed for 4 hours. The solvent was evaporated in vacuo and the solution poured into 40ml of water and extracted 3 times with 10ml of ether each time. The combined organic phases were washed with water and dried over magnesium sulfate. The solvent was distilled off to obtain a crystalline product. Melting point: 158-160 ℃ step E: 6-Methoxyphenanthrene-3, 4-dicarboxylic acid dimethyl ester

The product prepared in the previous step was dissolved in 150ml of dry toluene. 1.2 equivalents of dichlorodicyanoquinone were added and the solution was refluxed for 4 hours. After cooling, the solution was filtered and the filtrate was evaporated. The residue is then taken up in 50ml of water and the crude product is extracted 3 times with 10ml of ether each time. After concentration of the solution, the product is purified by chromatography on silica gel using CH₂Cl₂And (4) eluting. Melting point: 112-115 ℃ step F: 4-Methoxycarbonyl-6-methoxyphenanthrene-3-carboxylic acid methyl ester and 3-methoxycarbonyl-6-methoxyphenanthrene-4-carboxylic acid methyl ester

A solution of the product prepared in the previous step in 15ml of 1N sodium hydroxide and 20ml of methanol was refluxed for 3 hours. After cooling, the methanol is distilled off and the aqueous phase is acidified with 6N hydrochloric acid. The solution was extracted 3 times with 20ml of dichloromethane and the product was obtained after evaporation of the solvent. Step G: 10-methoxy-2-oxacyclopenta [ c ] phenanthrene-1, 3-dione

The mixture of acids prepared in the previous step was refluxed in 120ml of acetic anhydride for 24 hours. After cooling, the precipitate is filtered off, washed with ether and dried. Melting point: preparation of B at 238 ℃: 2-oxacyclopenta [ c ] phenanthrene-1, 3-dione step a: phenanthrene-3, 4-dicarboxylic acid dimethyl ester

A solution of 65mmol of 2-vinylnaphthalene and 68mmol of acetylenedicarboxylic acid methyl ester in 200ml of nitrobenzene was refluxed for 14 hours. The solvent is evaporated off and the residue is chromatographed on silica gel (eluent: CH)₂Cl₂) To obtain the desired product. Melting point: 117 ℃ step B: 3-methoxycarbonylphenanthrene-4-carboxylic acid and 4-methoxycarbonylphenanthrene-3-carboxylic acid

This mixture was prepared from the product of step a according to the method described in step F of preparation a. Melting point: 257 ℃ step C: 2-oxacyclopenta [ c ] phenanthrene-1, 3-dione

The desired product was prepared from the mixture of the previous step according to the method described in step G of preparation a. Melting point: preparation of C at 245 ℃: naphtho [2, 3-c ] furan-1, 3-dione

The desired product is prepared according to the method described in step G of preparation A, starting from naphthalene-2, 3-dicarboxylic acid. Preparation D: 2-oxa-8-thiadicyclopentadiene [ a, h ] naphthalene-1, 3-diones

The desired product is prepared according to the method described for preparation A, using 6, 7-dihydro-5H-benzo [ b ] thiophen-4-one in step A. Preparation E: 9-aza-9, 10-dimethyl-6-methoxy-2-oxa-9 IH-cyclopenta [ b ] fluorene-1, 3-dione step a: 1, 9-dimethyl-6-methoxy-9H-carbazole-2, 3-dicarboxylic acid dimethyl ester

The desired product was prepared from 1, 9-dimethyl-6-methoxy-9H-pyrano [3, 4-b ] indol-3-one according to the method described in chem. ber, 97, 667, 1964. And B: 1, 9-dimethyl-6-methoxy-2-methoxycarbonyl-9H-carbazole-3-carboxylic acid and 1, 9-dimethyl-6-methoxy-3-methoxycarbonyl-9H-carbazole-2-carboxylic acid

This mixture was prepared from the product described in step a according to the method described in step F of preparation a. And C: 9-aza-9, 10-dimethyl-6-methoxy-2-oxa-9H-cyclopenta [ b ] fluorene-1, 3-dione

The desired product is obtained according to the process described in step G of preparation A, starting from the compound described in the preceding step. Preparation F: 10-nitro-2-oxacyclopenta [ c ] phenanthrene-1, 3-dione step a: 6-Nitrofen-3, 4-dicarboxylic acid dimethyl ester

Ice-cold 38.8mmol of 70% nitric acid are added dropwise to a suspension of 32.3mmol of the product obtained in step A of preparation B in 120ml of trifluoroacetic anhydride cooled to 0 ℃. The solution was stirred at 0 ℃ for 3 hours and then at room temperature for 2 hours. The precipitate is filtered off, washed with diisopropyl ether and dried. Melting point: 180 ℃ (decomposition) step B: 6-Nitro-3-methoxycarbonylphenanthrene-4-carboxylic acid and 6-nitro-4-methoxycarbonylphenanthrene-3-carboxylic acid

This mixture was prepared according to the method described in step B of preparation G from the product obtained in the previous step. And C: 10-nitro-2-oxacyclopenta [ c ] phenanthrene-1, 3-dione

The desired product is obtained according to the process described in step G of preparation A, starting from the compound described in the preceding step. Preparation G: 9-oxacyclopenta [ b ] phenanthrene-8, 10-dione step a: naphthalen-1-ylacrylic acid ethyl ester

220ml of a 1N potassium tert-butoxide in tetrahydrofuran are added dropwise to a solution of 0.219mol of ethoxycarbonyltriphenylphosphonium bromide in 200ml of tetrahydrofuran. The solution was stirred at room temperature for 1 hour, and then a solution of 0.22mol of naphthalene-1-carbaldehyde in tetrahydrofuran was added dropwise. The solution was stirred at room temperature for 4 hours, filtered, the solvent was evaporated off and the residue was taken up in 400ml of dichloromethane. The organic phase is washed with 100ml of water, dried and concentrated. The residue was taken up in diisopropyl ether, the precipitate was filtered off and the solvent was concentrated to give the desired product. And B: naphthalen-1-yl acrylic acid

A solution of 0.2mol of the product prepared in the preceding step in 500ml of ethanol is stirred at room temperature in the presence of 250ml of 2N sodium hydroxide solution for 18 hours. The ethanol is evaporated off, the remaining solution is extracted with 100ml of ethyl acetate and the aqueous phase is acidified with 6N hydrochloric acid solution at 0 ℃. The desired product is filtered off and dried. Melting point: step C at 188 ℃: 2, 3-dibromonaphthalen-1-ylpropanoic acid

The desired product was prepared from the acid prepared in the previous step according to the method described in aust.j.chem., 16, 854, 1963. Melting point: 197 ℃ step D: naphthalen-1-ylpropynoic acid

A mixture of 0.1mol of the compound prepared in the previous step in 120ml of ethanol containing 0.36mol of potassium hydroxide was stirred under reflux for 2 hours and then at room temperature for 16 hours. The precipitate was filtered off and the filtrate was concentrated. The residue is taken up in ether and the precipitate is filtered off, washed with dichloromethane and then diluted with 400ml of water. The aqueous phase is acidified with concentrated hydrochloric acid, the desired product is filtered off, dried and recrystallized from carbon tetrachloride. Melting point: 95 ℃ step E: 9-oxacyclopenta [ b ] phenanthrene-8, 10-dione

The desired product was prepared from the compound prepared in the previous step according to the method described in aust.j.chem., 16, 854, 1963. Melting point: preparation of H at 278 ℃: benzo [ d ] benzo [2, 1-b; 3-4-c ] Furan-1, 3-dione step A: benzofuran-3-ylacetic acid methyl ester

A mixture of 0.4mol of coumaranone and 0.48mol of methyl (triphenylphosphoranylidene) acetate in 1 l of p-xylene is refluxed for 18 hours. After cooling to room temperature, the solvent is evaporated off, the residue is taken up in 1 l of ether, the precipitate is filtered off and washed with ether, the filtrate is concentrated and chromatographed on silica gel (eluent: CH)₂Cl₂) The desired product is obtained. And B: 2-benzofuran-3-ylethanol

0.38mol of the solution of the product prepared in the previous step was added to 0.64mol of LiAlH with stirring at 0 deg.C₄In a suspension of 1 liter of ether. The solution was stirred at room temperature for 1 hour, followed by dropwise addition of 100ml of ethyl acetate and 100ml of 1N hydrochloric acid in that order. After stirring for 18 hours, the solution was filtered and the filtrate was left to layer and the organic phase was washed with saturated NaCl solution and water in that order. The organic phase is dried and the solvent is distilled off to give the desired product. And C: 3- (2-bromoethyl) benzofuran

The desired product was prepared from the compound prepared in the previous step according to the method described in aust.j.chem., 44, 907, 1991. Step D: 3-vinylbenzofuran

The desired product was prepared from the compound prepared in the previous step according to the method described in aust.j.chem., 44, 907, 1991. Step E: 1, 2-Dihydrodibenzofuran-3, 4-dicarboxylic acid dimethyl ester

A1 liter degassed toluene solution containing 0.2mol of the compound prepared in the previous step and 0.2mol of acetylene dicarboxylic acid methyl ester was refluxed under an inert gas atmosphere for 24 hours. After concentration of the solvent, the mixture is chromatographed on silica gel (eluent CH)₂Cl₂) The desired product is obtained. Step F: dibenzofuran-3, 4-dicarboxylic acid dimethyl ester

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step G of preparation a. Melting point: step G at 123 ℃: dibenzofuran-3, 4-dicarboxylic acid

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step B of preparation G. Melting point: 260 ℃ and 262 ℃ step H: benzo [ d ] benzo [2, 1-b; 3-4-C ] difuran-1, 3-diones

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step G of preparation a. Melting point: preparation I at 258 deg.C: benzo [ d ] benzo [1, 2-b; 3-4-c ] Furan-1, 3-dione step A: 2-vinylbenzofuran

92ml of a 1N potassium tert-butoxide tetrahydrofuran solution are added dropwise at 0 ℃ to a vigorously stirred solution of 68.4mmol of benzofuran-2-carbaldehyde and 92mmol of methyltriphenylphosphonium bromide in 200ml of tetrahydrofuran and 150ml of dimethylformamide. The solution was stirred at room temperature for 2 hours, poured into 2 l of ice-water and extracted with ether. After concentrating the solvent, the residue is taken up in 1 l of petroleum ether, the precipitate is filtered off and washed with petroleum ether. The filtrate was evaporated to give the desired product. And B: dibenzofurane-1, 2-dicarboxylic acid dimethyl ester

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step E of preparation H. Melting point: step C at 126 ℃: dibenzofuran-1-methoxycarbonyl-2-carboxylic acid and dibenzofuran-2-methoxycarbonyl-1-carboxylic acid

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step B of preparation G. Melting point: 175-: benzo [ d ] benzo [1, 2-b; 3-4-C ] difuran-1, 3-diones

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step G of preparation a. Melting point: preparation of J at 211 ℃: 2-oxa-6-thiacyclopenta [ c ] fluorene-1, 3-dione step a: benzo [ b ] thiophene-2-carbaldehyde

The desired product was prepared from benzo [ b ] thiophene according to the procedures described in the American society for chemistry, 74, 2396, 1952. Melting point: step B at 40-41 ℃: vinyl-2-benzo [ b ] thiophenes

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step A of preparation I. Melting point: 132-: 3, 4-Dihydrodibenzothiophene-1, 2-dicarboxylic acid dimethyl ester

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step E of preparation H. Melting point: 129 ℃ step D: dibenzothiophene-1, 2-dicarboxylic acid dimethyl ester

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step E of preparation a. Step E: 1-methoxycarbonyl dibenzothiophene-2-carboxylic acid and

2-methoxycarbonyl dibenzothiophene-1-carboxylic acid

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step B of preparation G. Melting point: 248-250 ℃ step F: 2-oxa-6-thiacyclopenta [ c ] fluorene-1, 3-dione

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step G of preparation a. Melting point: preparation of K at 269 ℃: 10-methyl-10H-2-oxa-10-azacyclopenta [ a ] fluorene-1, 3-dione step a: 1-methyl-3-vinyl-1H-indoles

The desired product was prepared from 1-methyl-1H-indole-3-carbaldehyde according to the procedure described in step a of preparation I. And B: 9-methyl-4, 9-dihydro-3H-carbazole-1, 2-dicarboxylic acid dimethyl ester

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step E of preparation H. Melting point: 113 ℃ step C: 9-methyl-3H-carbazole-1, 2-dicarboxylic acid dimethyl ester

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step E of preparation a. Melting point: 139 ℃ step D: 9-methyl-1-methoxycarbonyl-3H-indole-2-carboxylic acid and

9-methyl-2-methoxycarbonyl-3H-indole-1-carboxylic acid

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step B of preparation G. Step E: 10-methyl-10H-2-oxa-10-azacyclopenta [ a ] fluorene-1, 3-dione

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step G of preparation a. Melting point: preparation of L at > 300 ℃: 6-methyl-6H-2-oxa-6-azacyclopenta [ c ] fluorene-1, 3-dione step A: (1-methyl-1H-indol-2-yl) methanol

The desired product was prepared from methyl 1-methyl-1H-indole-2-carboxylate according to the procedure described in step B of preparation H. Melting point: 104 ℃ step B: 1-methyl-1H-indole-2-carbaldehyde

A mixture of 0.1mol of the compound prepared in the preceding step, 1mol of manganese oxide and 0.37mol of sodium chloride in 500ml of ether is stirred at room temperature for 48 hours in a pressure vessel. The solution is filtered and concentrated and chromatographed on silica gel (eluent: CH)₂Cl₂) The desired product is obtained. Melting point: 83 ℃ step C: 1-methyl-2-vinyl-1H-indoles

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step A of preparation I. Step D: 9-methyl-2, 9-dihydro-9H-carbazole-3, 4-dicarboxylic acid dimethyl ester

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step E of preparation H. Melting point: 153 ℃ step E: 9-methyl-9H-carbazole-3, 4-dicarboxylic acid dimethyl ester

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step E of preparation a. Melting point: step F at 156 ℃: 9-methyl-9H-carbazole-3, 4-dicarboxylic acid

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step B of preparation G. Melting point: 294 ℃ step G: 6-methyl-6H-2-oxa-6-azacyclopenta [ c ] fluorene-1, 3-dione

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step G of preparation a. Melting point: preparation of M at > 300 ℃: 6-acetyl-6H-2-oxa-6-azacyclopenta [ c ] fluorene-1, 3-dione step A: (1H-indol-2-yl) methanol

The desired product was prepared from ethyl 1H-indole-2-carboxylate according to the procedure described in step B of preparation H. Melting point: 80 ℃ step B: 1H-indole-2-carbaldehyde

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step B for preparation L. And C: 2-vinyl-1H-indoles

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step A of preparation I. Melting point: 91 ℃ step D: 2, 9-dihydro-1H-carbazole-3, 4-dicarboxylic acid dimethyl ester

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step E of preparation H. Melting point: 249 ℃ step E: 1H-carbazole-3, 4-dicarboxylic acid dimethyl ester

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step E of preparation a. Melting point: 219 ℃ step F: 3-methoxycarbonyl-1H-carbazole-4-carboxylic acid and

4-methoxycarbonyl-1H-carbazole-3-carboxylic acid

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step B of preparation G. Step G: 6-acetyl-6H-2-oxa-6-azacyclopenta [ c ] fluorene-1, 3-dione

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step G of preparation a. Preparation of N: 6-methyl-9-nitro-6H-2-oxa-6-azacyclopenta [ c ] fluorene-1, 3-dione step A: 6-Nitro-9-methyl-9H-carbazole-3, 4-dicarboxylic acid dimethyl ester

The desired product was prepared according to the procedure described in step A of preparation F from the compound prepared in step E of preparation L. Melting point: 234 ℃ step B: 6-nitro-9-methyl-9H-carbazole-3, 4-dicarboxylic acid

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step F of preparation a. And C: 6-methyl-9-nitro-6H-2-oxa-6-azacyclopenta [ c ] fluorene-1, 3-dione

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step G of preparation a. Melting point: preparation 0 at > 300 ℃: 6-methyl-9-methoxy-6H-2-oxa-6-azacyclopenta [ c ] fluorene-1, 3-dione step A: 5-methoxy-1-methyl-1H-indolecarboxylic acid methyl ester

100ml of 1N potassium tert-butoxide in tetrahydrofuran are added dropwise to a solution of 0.1mol of methyl 5-methoxy-1H-indolecarboxylate in tetrahydrofuran which is cooled to-20 ℃. The reaction solution was stirred at this temperature for 30 minutes, followed by dropwise addition of a 0.1mol of methyl iodide in tetrahydrofuran. After the completion of the addition, the reaction solution was immediately warmed to room temperature, stirred at that temperature for 1 hour and then filtered. The desired product is obtained after concentration of the solvent. Melting point: step B at 200 ℃: (5-methoxy-1-methyl-1H-indol-2-yl) methanol

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step B of preparation H. Melting point: step C at 158 ℃: 5-methoxy-1-methyl-1H-indole-2-carbaldehyde

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step B for preparation L. Melting point: step D at 88-90 ℃: 5-methoxy-1-methyl-2-vinyl-1H-indole

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step A of preparation I. Melting point: 87-89 ℃ step E: 6-methoxy-9-methyl-2, 9-dihydro-1H-carbazole-3, 4-dicarboxylic acid dimethyl ester

The desired product was prepared from the compound prepared in the previous step according to the procedure described in step E of preparation H. Melting point: 245 ℃ step F: 6-methoxy-9-methyl-1H-carbazole-3, 4-dicarboxylic acid dimethyl ester

The desired product was prepared from the previous compound according to the procedure described in step E of preparation a. Melting point: 211 ℃ step G: 6-methoxy-9-methyl-1H-carbazole-3, 4-dicarboxylic acid

The desired product was prepared from the previous compound according to the procedure described in step F of preparation a. Step H: 9-methoxy-6-methyl-6H-2-oxa-6-azacyclopenta [ c ] fluorene-1, 3-dione

The desired product was prepared from the starting compound according to the procedure described in step G of preparation a. Preparation of P: 5, 6-dimethyl-6H-2-oxa-6-azacyclopenta [ c ] fluorene-1, 3-dione step A: 2-isopropenyl-1-methyl-1H indoles

The desired product was prepared from N-methyl-1H indole according to the method described in journal of organic chemistry, 59, (15), 4250, 1994. And B: 1, 9-dimethyl-1, 2-dihydro-9H-carbazole-3, 4-dicarboxylic acid dimethyl ester

The desired product was prepared from the product of step A according to the procedure described in step E of preparation H. And C: 1, 9-dimethyl-9H-carbazole-3, 4-dicarboxylic acid dimethyl ester

The desired product was prepared from the above product according to the procedure described in step E of preparation A. Step D: 1, 9-dimethyl-9H-carbazole-3, 4-dicarboxylic acid

The desired product was prepared from the previous compound according to the procedure described in step F of preparation a. Step E: 5, 6-dimethyl-6H-2-oxa-6-azacyclopenta [ c ] fluorene-1, 3-dione

The desired product was prepared from the starting compound according to the procedure described in step G of preparation a. Preparation of Q: n, N-bis [2- (1(R) -methylaminoethyl) ethane ] -1, 2-diamine

The desired product was obtained from N- (tert-butoxycarbonyl) -D-alanine and 1, 2-ethylenediamine according to the method described in journal of pharmaceutical chemistry, 40, 449, 1997. Example 1: n, N-bis [2- (2-aza-1, 3-dioxo-10-methoxy-cyclopenta [ c ] phenanthrene-2-

Yl) Ethyl ] ethane-1, 2-diamine dimesylate

40mmol of N-1- [2- (2-aminoethylamino) ethyl ] ethane-1, 2-diamine in 20ml of toluene are added dropwise to 80mmol of a 1 l toluene solution of the compound mentioned in preparation A, and stirred at 80 ℃. The reaction mixture was then refluxed for 17 hours, filtered while hot and concentrated under reduced pressure. The residue was taken up in 300ml of ethanol, stirred under reflux for 3 hours and filtered hot. After cooling to room temperature, the crude product was isolated by filtration and then purified by silica gel chromatography. The collected base form product was stirred in 450ml dichloromethane. To this solution was added two equivalents of methanesulfonic acid in dichloromethane and stirring was continued for 5 hours. The product is filtered off, washed with ether and dried. Example 2: n, N-bis [2- (2-aza-1, 3-dioxo-10-methoxy-cyclopenta [ c ] phenanthrene-2-

Yl) Ethyl ] propane-1, 3-diamine dimesylate

The desired product was prepared according to the procedure described in example 1, using the compound described in preparation A and N-1- [3- (2-aminoethylamino) propyl ] ethane-1, 2-diamine. Example 3: 1, 8-bis (2-aza-1, 3-dioxo-10-methoxy-cyclopenta [ c ] phenanthrene-2-

4-azaoctane methanesulfonate

The desired product was prepared according to the procedure described in example 1, using the compound described in preparation A and 4-azaoctane-1, 8-diamine. Melting point: 294 ℃ example 4: n, N-bis [3- (2-aza-1, 3-dioxo-10-methoxy-cyclopenta [ c ] phenanthrene-2-

Methyl) propyl methylamine methanesulfonate the desired product was prepared according to the procedure described in example 1, using the compound described in preparation A and N, N-bis (3-aminopropyl) methylamine. Example 5: n, N-bis [2- (2-aza-1, 3-dioxocyclopenta [ c ] phenanthren-2-yl) ethyl ] -

Propane-1, 3-diamine disulfonate

The desired product was prepared according to the procedure described in example 1, using the compound described in preparation B and N-1- [3- (2-aminoethylamino) propyl ] ethane-1, 2-diamine. Analyzing trace elements:

c% H% N% S% calculated 60.584.966.897.89 found 60.794.916.837.79 example 6: n, N-bis [2- (2-aza-1, 3-dioxo-cyclopenta [ c ] phenanthren-2-yl) ethyl ] -

Ethane-1, 2-diamine disulfonate

The desired product was prepared according to the procedure described in example 1, substituting the product described in preparation B for the product described in preparation A. Analyzing trace elements:

c% H% N% S% calculated 60.144.797.018.03 found 60.324.896.978.02 example 7: n, N-bis [2- (2-aza-1, 3-dioxo-naphtho [2, 3-c ] furan-2-yl) ethyl ] -

Ethane-1, 2-diamine disulfonate

The desired product was prepared according to the procedure described in example 1, substituting the product described in preparation C for the product described in preparation A. Analyzing trace elements:

c% H% N% S% calculated 55.014.908.029.18 found 54.404.977.778.32 example 8: n, N-bis [2- (2-aza-1, 3-dioxo-8-thiadicyclopentadiene [ a, h ] naphthalene-2-

Yl) Ethyl ] ethane-1, 2-diamine dimesylate

The desired product was prepared according to the procedure described in example 1, substituting the product described in preparation D for the product described in preparation A. Analyzing trace elements:

c% H% N% S% calculated 45.504.225.5919.18 found 45.964.115.6319.88 example 9: n, N-bis [3- (2-aza-1, 3-dioxo-8-thiadicyclopentadiene [ a, h ] naphthalene-2-

Methyl) propyl methylamine methanesulfonate the desired product was prepared according to the procedure described in example 1, using the compound described in preparation D and N, N-bis (3-aminopropyl) methylamine. Example 10: n, N-bis [2- (2, 9-diaza-9, 10-dimethyl-1, 3-dioxo-6-methoxy-)

9H-cyclopenta [ b ] fluoren-2-yl) ethyl ] ethane-1, 2-diamine disulfonate

The desired product was prepared according to the procedure described in example 1, substituting the product described in preparation E for the product described in preparation A and the reaction solvent with ethanol for toluene. Analyzing trace elements:

c% H% N% S% calculated 56.715.429.417.18 found 56.715.619.517.11 example 11: n, N-bis [2- (2, 9-diaza-9, 10-dimethyl-1, 3-dioxo-6-methoxy-)

9H-cyclopenta [ b ] fluoren-2-yl) ethyl ] propane-1, 3-diamine disulfonate

The desired product was prepared according to the procedure described in example 1, starting from the compound described in preparation E and N-1- [3- (2-aminoethylamino) propyl ] ethane-1, 2-diamine and replacing the reaction solvent with ethanol for toluene. Analyzing trace elements:

c% H% N% S% calculated 56.945.569.277.02 found 57.495.619.357.13 example 12: n, N-bis [3- (2, 9-diaza-9, 10-dimethyl-1, 3-dioxo-6-methoxy-9H-)

Cyclopenta [ b ] fluoren-2-yl) propyl ] methylamine methanesulfonate

The desired product was prepared according to the procedure described in example 1, starting from the compound described in preparation E and N, N-bis (3-aminopropyl) methylamine and replacing the reaction solvent with ethanol for toluene. Analyzing trace elements:

c% H% N% S% calculated 63.385.708.804.03 found 64.195.829.184.03 example 13: 1, 8-bis (2, 9-diaza-9, 10-dimethyl-1, 3-dioxo-6-methoxy-9H-ring

Pentadieno [ b ] fluoren-2-yl) -4-azaoctane methanesulfonate

The desired product was obtained according to the procedure described in example 1, starting from the compound described in preparation E and 4-azaoctane-1, 8-diamine and replacing the toluene by ethanol as reaction solvent. Analyzing trace elements:

c% H% N% S% calculated 63.385.708.804.03 found 62.955.808.964.02 example 14: n, N-bis [3- (2, 9-diaza-9, 10-dimethyl-1, 3-dioxo-6-methoxy-9H-)

Cyclopenta [ b ] fluoren-2-yl) propyl ] amine methanesulfonate

The desired product was prepared according to the procedure described in example 1, starting from the compound described in preparation E and N, N-bis (3-aminopropyl) amine and replacing the reaction solvent with ethanol for toluene. Analyzing trace elements:

c% H% N% S% calculated 62.985.548.964.10 found 62.915.638.784.17 example 15: n, N-bis [3- (2, 9-diaza-9, 10-dimethyl-1, 3-dioxo-6-methoxy-)

9H-cyclopenta [ b ] fluoren-2-yl) propyl ] ethane-1, 2-diamine disulfonate

The desired product is obtained according to the method described in example 1, starting from the compound described in preparation E and N- [2- (3-aminopropylamino) ethyl ] propane-1, 3-diamine and replacing the toluene by ethanol as reaction solvent. Analyzing trace elements:

c% H% N% S% calculated 57.385.699.126.96 found 57.085.668.646.50 example 16: n, N-bis [2- (2-aza-1, 3-dioxo-10-hydroxycyclopenta [ c ] phenanthren-2-yl)

Ethyl ] ethane-1, 2-diamine hydrobromide salt

A suspension of 1mmol of the compound from example 1 in 40ml of 47% hydrobromic acid is refluxed for 8 hours. After cooling to room temperature, the solid was filtered off, washed with water and refluxed in ethanol for 1 hour. The desired product is obtained after filtration. Example 17: n, N-bis [3- (2-aza-1, 3-dioxo-10-hydroxycyclopenta [ c ] phenanthren-2-yl)

Propyl methylamine hydrobromide

The desired product was prepared according to the procedure described for example 16, starting from the compound of example 4. Example 18: n, N-bis [3- (2, 9-diaza-9, 10-dimethyl-1, 3-dioxo-6-hydroxy-9H-)

Cyclopenta [ b ] fluoren-2-yl) ethyl ] ethane-1, 2-diamine dihydrobromide salt

The desired product was prepared according to the procedure described for example 16, starting from the compound of example 10. Example 19: n, N-bis [2- (2, 9-diaza-9, 10-dimethyl-1, 3-dioxo-6-hydroxy-9H-)

Cyclopenta [ b ] fluoren-2-yl) ethyl ] propane-1, 3-diamine dihydrobromide salt

The desired product was prepared according to the procedure described for example 16, starting from the compound of example 11. Analyzing trace elements:

calcd for C% H% N% Br% 55.204.759.9018.83 found 55.114.449.7317.53 example 20: 1, 8-bis (2, 9-diaza-9, 10-dimethyl-1, 3-dioxo-6-hydroxy-9H-cyclopenta-lene

Dieno [ b ] fluoren-2-yl) -4-azocane hydrobromide

The desired product was prepared according to the procedure described for example 16, starting from the compound of example 13. And (3) dividing the trace elements into:

calcd for C% H% N% Br% 62.245.099.3010.62 found 61.045.029.259.57 example 21: n, N-bis [3- (2, 9-diaza-9, 10-dimethyl-1, 3-dioxo-6-hydroxy-9H-)

Cyclopenta [ b ] fluoren-2-yl) propyl ] ethane-1, 2-diamine dihydrobromide salt

The desired product was prepared according to the procedure described for example 16, starting from the compound of example 15. Analyzing trace elements:

calcd for C% H% N% Br% 55.704.919.7418.53 found 55.884.689.1317.87 example 22: 1- [2- (1, 3-dioxo-2, 3-dihydro-5-nitrobenzo [ d, e ] isoquinolin-2-yl) ethyl ester

3- [2- (2-aza-1, 3-dioxo-10-methoxy-cyclopenta [ c ] phenanthrene-

2-yl) ethyl ] propane-1, 3-diamine disulfonate step a: 2- [2- (3-aminopropylamino) ethylamino ] -5-nitrobenzo [ d, e ] isoquinoline-1, 3-

Diketone trimethosulfonate

A solution of 50mmol of 3-nitronaphthoic anhydride in 300ml of THF at 0 ℃ is added dropwise to a stirred solution of 50mmol of N, N-bis (2-aminoethyl) propane-1, 3-diamine in 200ml of THF at 0 ℃. After the addition was complete, the reaction mixture was stirred at room temperature for 3 hours, under reflux for 2 hours and then filtered while hot. The filtrate is stirred at room temperature and a solution of 150mmol of methanesulfonic acid in 100ml of THF is added dropwise. The precipitate is filtered off, added to 1 l of ethanol and refluxed for 2 hours. The insoluble material was filtered off and the filtrate was cooled. Filtration gave the desired product in 32% yield. And B: 1- [2- (1, 3-dioxo-2, 3-dihydro-5-nitrobenzo [ d, e ] isoquinolin-2-yl) ethyl ester

3- [2- (2-aza-1, 3-dioxo-10-methoxy-cyclopenta [ c ] phenanthrene-2-

Yl) Ethyl ] propane-1, 3-diamine dimesylate

5mmol of the compound of preparation A were added portionwise, with stirring at room temperature, to a solution containing 5mmol of the compound of preparation A prepared in the preceding stepCompound and 15mmo l diisopropylethylamine in 1 l ethanol. After the addition was complete, the mixture was refluxed for 15 hours and then filtered while hot. The cooled filtrate was filtered and the solid was purified by chromatography on silica gel using 95 CH₂Cl₂/5 MeOH/0.1 NH₂OH as eluent. The fractions containing the product are combined and concentrated, the residue is taken up in 1 l of dichloromethane and 2 equivalents of methanesulfonic acid are added. The product was isolated by filtration. Example 23: 1- [2- (1, 3-dioxo-2, 3-dihydro-5-nitrobenzo [ d, e ]]Isoquinolin-2-yl) ethyl

2- [2 (2-aza-1, 3-dioxo-10-methoxy-cyclopenta [ c ] phenanthrene-

2-yl) ethyl ] ethane-1, 2-diamine disulfonate

The desired product was prepared according to the procedure described for example 22, using N, N-bis (2-aminoethyl) ethane-1, 2-diamine in step A. Analyzing trace elements:

c% H% N% S% calculated 53.004.528.318.37 found 53.004.648.408.54 example 24: n, N-bis [2- (2-aza-6-oxacyclopenta [ c ] fluorene-1, 3-dione-2-yl)

Ethyl propane-1, 3-diamine disulfonate

The desired product was prepared according to the procedure described in example 1 from the compound described in preparation I and N-1- [3- (2-aminoethylamino) propyl ] ethane-1, 2-diamine. Analyzing trace elements:

c% H% N% S% calculated 56.054.587.078.09 found 56.154.487.028.30 example 25: n, N-bis [2- (2-aza-1, 3-dioxo-10-methoxy-cyclopenta [ c ] phenanthrene-

2-yl) ethyl propane-1, 3-diamine disulfonate

The desired product was prepared according to the procedure described in example 1 from the compound described in preparation F and N-1- [3- (2-aminoethylamino) propyl ] ethane-1, 2-diamine. Analyzing trace elements:

c% H% N% S% calculated 54.544.249.317.10 found 54.734.279.247.24 example 26: n, N-bis [2- (2-aza-10-oxa-1, 3-dioxo-cyclopenta [ a ] fluorene-2-

Yl) Ethyl ] propane-1, 3-diamine dimesylate

The desired product was prepared according to the procedure described in example 1 from the compound described in preparation H and N-1- [3- (2-aminoethylamino) propyl ] ethane-1, 2-diamine. Analyzing trace elements:

c% H% N% S% calculated 56.054.587.078.09 found 56.484.667.037.85 example 27: n, N-bis [2- (2-aza-1, 3-oxo-10-methoxy-cyclopenta [ c ] phenanthrene-

2-Yl) (1(R) -methyl) ethyl ] ethane-1, 2-diamine disulfonate

The desired product was prepared according to the procedure described in example 1 from the compound described in preparation A and the compound described in preparation Q. Analyzing trace elements: old fashioned C% H% N% S% calculated 59.585.236.327.23 found 59.775.266.156.99 example 28: n, N-bis [2- (2-aza-1, 3-dioxo-cyclopenta [ b ] phenanthren-2-yl) ethyl ]

Propane-1, 3-diamine disulfonate

The desired product was prepared according to the procedure described in example 1 from the compound described in preparation G and N-1- [3- (2-aminoethylamino) propyl ] ethane-1, 2-diamine. Example 29: n, N-bis [2- (2-aza-1, 3-dioxo-10-methyl-10H pyrrolo [3, 4-a ] carbo [ ]

Azol-2-yl) ethyl ] propane-1, 3-amine dimesylate

The desired product was prepared according to the procedure described in example 1 from the compound described in preparation K and N-1- [3- (2-aminoethylamino) propyl ] ethane-1, 2 diamine. Analyzing trace elements:

c% H% N% S% calcd 57.205.1710.267.83 found 57.185.099.928.04 example 30: n, N-bis [2- (2-aza-1, 3-dioxo-6-thiacyclopenta [ c ] fluoren-2-yl)

Ethyl propane-1, 3-diamine disulfonate

The desired product was prepared according to the procedure described in example 1 from the compound described in preparation J and N-1- [3- (2-aminoethylamino) propyl ] ethane-1, 2-diamine. Analyzing trace elements:

c% H% N% S% calculated 53.874.406.7915.55 found 53.264.816.8815.88 example 31: n, N-bis [2- (2-aza-1, 3-dioxo-6-methyl-6H-pyrrolo [3, 4-c ] carbazole

-2-yl) ethyl ] propane-1, 3-diamine disulfonate

The desired product was prepared according to the procedure described in example 1 from the compound described in preparation L and N-1- [3- (2-aminoethylamino) propyl ] ethane-1, 2-diamine. Analyzing trace elements:

c% H% N% S% calculated 57.205.1710.267.83 found 57.185.099.928.04 example 32: n, N-bis [2- (2-aza-1, 3-dioxo-10-methoxy-cyclopenta [ c ] phenanthrene-

2-yl) ethyl butane-1, 4-diamine disulfonate

The desired product was prepared according to the procedure described in example 1 from the compound described in preparation A and N-1- [4- (2-aminoethylamino) butyl ] ethane-1, 2-diamine. Analyzing trace elements:

c% H% N% S% calculated 59.585.236.327.23 found 59.175.376.266.98 example 33: n, N-bis [2- (2-aza- ], 3-dioxo-6H-pyrrolo [3, 4-c ] carbazol-2-yl)

Ethyl propane-1, 3-diamine disulfonate

The desired product was prepared according to the procedure described in example 1 from the compound described for preparation M and N-1- [3- (2-aminoethylamino) propyl ] ethane-1, 2-diamine. Analyzing trace elements:

c% H% N% S% calculated 56.194.8410.638.11 found 56.345.0810.718.24 example 34: 1- [2- (1, 3-dioxo-2, 3-dihydro-5-nitrobenzo [ d, e ] isoquinolin-2-yl) ethyl ester

3- [2- (2-aza-1, 3-dioxo-6-methyl-6H-pyrrolo [3, 4-c ] carbazole

-2-yl) ethyl ] propane-1, 3-diamine disulfonate

The desired product was prepared according to the procedure described for example 22, using the compound described for preparation L in step B. Analyzing trace elements:

c% H% N% S% calculated 53.334.7210.367.91 found 53.804.7010.467. S8 example 35: n, N-bis [2- (2-aza-1, 3-dioxo-6-methyl-9-methoxy-6H-pyrrolo [)

[3, 4-c ] carbazole) -2-yl ] propane-1, 3-diamine disulfonate

The desired product was prepared according to the procedure described in example 1 using the compound described for preparation O and N-1- [3- (2-aminoethylamino) propyl ] ethane-1, 2-diamine. Example 36: n, N-bis [2- (2-aza-1, 3-dioxo-6-methyl-6H-pyrrolo [3, 4-c ] carbazole

-2-yl) (1- (R) -methyl) ethyl ] ethane-1, 2-diamine disulfonate

The desired product was prepared according to the procedure described in example 1, using the compound described for preparation L and the compound described for preparation Q. Example 37: 1- [2- (1, 3-dioxo-2, 3-dihydro-5-nitrobenzo [ d, e ] isoquinoline-2-)

Radical) (1- (R) -methyl) ethyl ] -2- [2- (2-aza-1, 3-dioxo-6-methyl-6H-

Pyrrolo [3, 4-c ] carbazol-2-yl) (1- (R) -methyl) ethyl ] ethane-1, 2-diamine

Dimethyl sulfonate

The desired product was prepared according to the procedure described for example 22, using the compound described for preparation L in step B. Example 38: n, N-bis [2- (2-aza-1, 3-dioxo-6-methyl-6H-9-nitropyrrolo)

[3, 4-c ] carbazol-2-yl) (1- (R) -methyl) ethyl ] ethane-1, 2-diamine dimethyl

Sulfonate salt

The desired product was prepared according to the procedure described in example 1, using the compound described in preparation N. Example 39: n, N-bis [2- (2-aza-5, 6-dimethyl-1, 3-dioxo-6-methyl-6H-pyrrolo [)

[3, 4-c ] carbazol-2-yl) (1- (R) -methyl) ethyl ] ethane-1, 2-diamine dimethylsulfonic acid

Acid salts

The desired product was prepared according to the procedure described in example 1, using the compound described in preparation P. Example 40: 1- [2- (2-aza-1, 3-dioxo-6-methyl-6H-pyrrolo [3, 4-c ] carbazole-2-

Radical) (1- (R) -methyl) ethyl ] - [2- (2-aza-1, 3-dioxo-10-methoxy-

Cyclopenta [ c ] phenanthren-2-yl) (1- (R) -methyl) ethyl ] ethane-1, 2-diamine

Dimethyl sulfonate

The desired product was prepared according to the procedure described for example 22, using the compound described in preparation A in step A and the compound described in preparation L in step B.

Pharmacological studies of the compounds of the invention example 41: proliferation in vitro

In this assay, the concentration at which the product inhibits cell growth by 50% (IC) can be determined by comparison with untreated control cells₅₀) To determine the in vitro antiproliferative capacity of the compounds.

The assay is based on a split-colorimetric assay in which soluble tetrazolium salt, 3- (4, 5-dimethylthiazol-2-yl) -2, 5-diphenyltetrazolium bromide (MTT), is reduced to medium-insoluble crystals of purple formazan * by the mitochondrial succinate dehydrogenase of living cells.

To ensure complete cell attachment, cells were seeded in 96-well plates at 625 cells/well and incubated continuously after 24 hours at 37 ℃ in the presence of a defined concentration range of test product. After four passages of doubling of the cell number (as measured by growth kinetics of the cell line), the live cells were stained with MTT (one-tenth the volume of the culture medium added to the wells in PBS containing 5mg/ml MTT) for 4 hours at 37 ℃. At the end of the incubation, a solution of SDS (20% SDS, 50% aqueous dimethylformamide, pH4.7) was added to the wells in the same volume, and after overnight contact, the solution dissolved crystals of formazan *. The optical density of the medium was then determined with a Titertek Multi-Scan MCC plate reader (Labsystem) at 540 nm. Calibration curves for each cell line were obtained by inoculating cells at increasing densities and a linear relationship between viable cell count and optical density readings was established by a standard curve.

The cell lines used in this assay were:

LLC cell line: from murine Lewis lung carcinoma.

-HT-29 cell line: from human colon cancer.

The results obtained from the above two cell lines indicate that the compounds of the invention have very strong antiproliferative capacity.

For exampleIC of the Compound of example 22₅₀Equal to 53.60nM in LLC line and 1.4nM in HT-29 line. Example 42: antitumor Activity in vivo

The antitumor activity of the compounds of the present invention was studied in nude mice. Fragments of human epidermal cancer KB-3-1 were subcutaneously transplanted into female Swiss nude mice of 4 to 6 weeks of age and a body weight of 20 to 22 g. When the tumor diameter reached approximately 6mm, the mice were divided into control and treatment groups (7 mice per group) to obtain the same average tumor volume in each group. Administering the test product by intravenous route; the injection is administered once daily for 5 days. Tumors were measured twice weekly and tumor volumes were calculated according to the following formula: volume (length x width)²)/2. Antitumor activity was assessed by two parameters: average T/C, expressed as a percentage; and SGD (specific growth retardation). Mean T/C (%) at each time point T is 100 × (Vt/V0) treatment mean/(V0/Vt) control mean, where Vt: tumor volume at time point t; v0: tumor volume at the beginning of treatment. SGD ═ Td treatment-Td control)/Td control, where Td ═ mean tumor doubling time. The results obtained with the products of the invention show that they strongly inhibit the growth of tumors.

For example, the compound of example 1, administered by intravenous route at a dose of 15mg/kg for 5 days, gave the following results: 14 days after treatment initiation, the mean T/C was 24% (i.e., 74% inhibition) and SGD was 1.3. Furthermore, the treatment did not result in any weight loss, so the compound was well tolerated. Example 43: pharmaceutical composition

The. ...3g

Claims (14)

1. A compound of formula (I)Wherein: m and n, which may be the same or different, represent 0 or 1, Z and Y, which may be the same or different, represent hydrogen, a halogen atom or a linear or branched (C)₁-C₆) Alkyl, straight-chain or branched (C)₁-C₆) Trihaloalkyl, straight or branched (C)₁-C₆) Alkoxy, hydroxy, cyano, nitro, amino, alkylamino or dialkylamino, Z represents a linear or branched C₄To C₁₂Alkylene radicalChain, one or more-CH therein₂-the group is optionally replaced by any one of the following atoms or groups: -NR- (wherein R represents a hydrogen atom or a linear or branched (C)₁-C₆) Alkyl), -O-, -S-, -SO-, -SO-₂-or-CONH-, or a substituted or unsubstituted heterocyclic group, a forms with two adjacent carbon atoms of the phenyl ring:

-a substituted or unsubstituted benzene ring,

-a substituted or unsubstituted naphthalene ring,

-a substituted or unsubstituted tetrahydronaphthalene ring or a substituted or unsubstituted 1, 4-dioxo-1, 2, 3, 4-tetrahydronaphthalene ring,

-or a substituted or unsubstituted heterocyclic ring,represents any one of the following groups:wherein: m is₁And n₁Which may be identical or different, represent 0 or 1, Z₁And Y₁Which may be the same or different, represent hydrogen, a halogen atom or a linear or branched (C)₁-C₆)

Alkyl, straight-chain or branched (C)₁-C₆) Trihaloalkyl, straight-chain or branched

(C₁-C₆) Alkoxy, hydroxy, cyano, nitro, amino, alkylamino or dioxane

Alkylamino radical, A₁Formed with two adjacent carbon atoms on the phenyl ring:

-a substituted or unsubstituted benzene ring,

-a substituted or unsubstituted naphthalene ring,

-a substituted or unsubstituted tetralin ring or a substituted or unsubstituted 1, 4-dioxygen

-a 1, 2, 3, 4-tetrahydronaphthalene ring,

-a substituted or unsubstituted heterocyclic ring,

wherein Z₂And Y₂May be the same or different and represents hydrogen, a halogen atom or a linear or branched (C)₁-C₆) Alkyl, straight-chain or branched (C)₁-C₆) Trihaloalkyl, straight or branched (C)₁-C₆) Alkoxy, hydroxy, cyano, nitro, amino, alkylamino or dialkylamino, isomers thereof and addition salts thereof with a pharmaceutically acceptable acid or base.

2. The compound of formula (I) as claimed in claim 1, wherein a forms a substituted or unsubstituted naphthalene ring with two adjacent carbon atoms of the benzene ring.

3. A compound of formula (I) as claimed in claim 1 wherein a forms with two adjacent carbon atoms of the phenyl ring a substituted or unsubstituted heterocyclic ring, preferably selected from the group consisting of optionally substituted indole, optionally substituted benzo [ b ] thiophene and optionally substituted benzo [ b ] furan.

4. A compound of formula (I) according to claim 1, wherein Z represents C₄To C₁₂Alkylene chain, wherein 1, 2 or 3-CH's in the chain₂The radical being substituted by 1, 2 or 3-NR-radicals (in which R represents a hydrogen atom or a linear or branched (C)₁-C₆) Alkyl) is substituted.

5. A compound of formula (I) according to claim 1, whereinRepresentsWherein:

m₁and n₁Which may be identical or different, represent 0 or 1,

Z₁and Y₁Which may be the same or different, represent hydrogen, a halogen atom or a linear or branched (C)₁-C₆)

Alkyl, straight-chain orBranched (C)₁-C₆) Trihaloalkyl, straight-chain or branched

(C₁-C₆) Alkoxy, hydroxy, cyano, nitro, amino, alkylamino or dioxane

An amino group as a radical,

A₁formed with two adjacent carbon atoms on the phenyl ring:

-a substituted or unsubstituted benzene ring,

-a substituted or unsubstituted naphthalene ring,

-a substituted or unsubstituted tetralin ring or a substituted or unsubstituted 1, 4-dioxygen

-a 1, 2, 3, 4-tetrahydronaphthalene ring,

-substituted or unsubstituted heterocycles, their isomers and their addition salts with pharmaceutically acceptable acids or bases.

6. A compound of formula (I) according to claim 1, whereinRepresents

Wherein Z₂And Y₂May be the same or different and represents hydrogen, a halogen atom or a linear or branched (C)₁-C₆) Alkyl, straight-chain or branched (C)₁-C₆) Trihaloalkyl, straight or branched (C)₁-C₆) Alkoxy, hydroxy, cyano, nitro, amino, alkylamino or dialkylamino, isomers thereof and addition salts thereof with a pharmaceutically acceptable acid or base.

7. A compound of formula (I) according to claim 1, wherein m and n simultaneously represent 0.

8. A compound of formula (I) according to claim 1: n, N-bis [2- (2-aza-1, 3-dioxo-10-methoxy-cyclopenta [ c ] phenanthren-2-yl) ethyl ] ethane-1, 2-diamine and addition salts thereof with a pharmaceutically acceptable acid.

9. A compound of formula (I) according to claim 1: n, N-bis [2- (2-aza-1, 3-dioxo-10-methoxy-cyclopenta [ c ] phenanthren-2-yl) ethyl ] propane-1, 3-diamine and addition salts thereof with a pharmaceutically acceptable acid.

10. A compound of formula (I) according to claim 1: 1- [2- (1, 3-dioxo-2, 3-dihydro-5-nitrobenzo [ d, e ] isoquinolin-2-yl) ethyl ] -3- [2- (2-aza-1, 3-dioxo-10-methoxy-cyclopenta [ c ] phenanthren-2-yl) ethyl ] propane-1, 3-diamine and its addition salts with a pharmaceutically acceptable acid.

11. A compound of formula (I) according to claim 1: 1- [2- (1, 3-dioxo-2, 3-dihydro-5-nitrobenzo [ d, e ] isoquinolin-2-yl) ethyl ] -2- [2- (2-aza-1, 3-dioxo-10-methoxy-cyclopenta [ c ] phenanthren-2-yl) ethyl ] ethane-1, 2-diamine and addition salts thereof with a pharmaceutically acceptable acid.

12. A process for the preparation of a compound of formula (I) as claimed in claim 1, using an anhydride of formula (II):wherein A, Z, Y, m and n are as defined for formula (I), with an excess of a diamine of formula (III):

H₂N-Z-NH₂(III) wherein Z is as defined for formula (I) and is isolated to yield: -a compound of formula (I/a), a particular case of compounds of formula (I):wherein A, Z, Y, m and n have the same meanings as in formula (I), -or (IV):thereinA, Z, Y, m and n have the same meaning as in formula (I), -subjecting it to the following reaction:

reacting with an anhydride of formula (II/a):wherein A is₁，Z₁，Y₁，m₁And n₁As defined for formula (I) to give a compound of formula (I/b) -a particular case of compounds of formula (I):wherein A, Z, Y, m, n, Z, A₁，Z₁，Y₁，m₁And n₁Has the same meaning as in formula (I),

or with an anhydride of formula (V):wherein Z₂And Y₂As defined for formula (I) to give a compound of formula (I/c) -a particular case of compounds of formula (I):wherein A, Z, Y, Z₂，Y₂And Z, m and n have the same meanings as in formula (I), and if desired, the compounds of formula (I/a), (I/b) or (I/c) can be subjected to the standard reaction for converting the substituents on the aromatic ring and, where appropriate, purified according to standard purification techniques, the isomers of which can be selectively separated according to standard separation techniques and, if desired, converted into addition salts thereof with a pharmaceutically acceptable acid or base.

13. A pharmaceutical composition comprising as active ingredient at least one compound as claimed in any one of claims 1 to 11, which composition comprises the active ingredient alone or in combination with one or more inert, non-toxic pharmaceutically acceptable excipients.

14. A pharmaceutical composition as claimed in claim 13, containing at least one compound as claimed in any one of claims 1 to 11 as an active ingredient, which is useful as an anticancer agent.