WO2000053166A2

WO2000053166A2 - Use of condensed 2,3-benzodiazepine derivatives for the treatment and prophylaxis of diseases related to the inhibition of lipid peroxidation

Info

Publication number: WO2000053166A2
Application number: PCT/HU2000/000011
Authority: WO
Inventors: Judit RÁCZNÉ BAJNÓGEL; Gábor SZÉNÁSI; Gábor Gigler; György Lévay; Géza SZABÓ; Károly TIHANYI; András Egyed; József Barkóczy; Zoltán RÁTKAI; Zoltán Greff; Géza Schneider; Gyula Simig; László BALÁZS; Imre Domán; Péter KÓTAY NAGY; Péter SERES
Original assignee: EGIS Gyógyszergyár Rt.
Priority date: 1999-02-15
Filing date: 2000-02-15
Publication date: 2000-09-14
Also published as: WO2000053166A3; HUP9900354A3; AU2819400A; HUP9900354A2; HU9900354D0

Abstract

The invention relates to the use of 8-substituted-9H-1,3-dioxolo(4,5-h)(2,3)benzodiazepine derivatives of general formula (I) and pharmaceutically acceptable acid addition salts and quaternary ammonium derivatives thereof for the preparation of pharmaceutical compositions suitable for the treatment or prophylaxis of diseases and states related to increased (pathological) lipid peroxidation and diseases caused by increased production of free radicals, e.g. atherosclerosis, pancreatitis, rheumatoid arthritis, cataract, Parkinson's disease, inflammatory diseases (e.g. hepatitis), ischemic heart disease, cerebral ischemia and renal failure.

Description

Use of condensed 2.3-benzodiazep-ne derivatives for the treatment and prophylaxis of diseases related to the inhibition of lipid peroxidation

Technical field of the invention

This invention relates to the use of 8-substituted-9H- 1,3-dioxolo[4,5-h][2,3]benzodiazepine derivatives for the treatment and prophylaxis of diseases and states related to the inhibition of lipid peroxidation and to the preparation of pharmaceutical compositions suitable for such treatment. Background of the invention

Several 2,3-benzodiazepine derivatives having biological activity are known.

Tofisopam i.e. 1-(3,4-dimethoxyphenyl)-5-ethyl-7,8- -dimethoxy-4-methyl-5H-2,3-benzodiazepine having anxiolytic effect is known form HU-P No. 155 572 and GB-P No. 1 202 579, respectively. The known compound does not comprise the ring system 1 ,3-dioxolo[4,5-h][2,3]benzodiazepine.

From HU-P No. 186 760, 7,8-dihydro-8-methyl-9H-1,3- -dioxolo[4,5-h][2,3]benzodiazepine derivatives having effect on the central nervous system are known, among others. The known compounds are prepared by reducing the corresponding 8-methyl-9H-1,3-dioxolo[4,5-h][2,3]benzodiazepine derivative.

Various substituted 8-methyl-9H-1 ,3-dioxolo[4,5- -h][2,3]benzodiazepine derivatives are known from HU-P No. 191 698 and the corresponding GB-P No. 2 162 184. The known compounds have antiaggressive and anxiolytic activities.

A novel process for the preparation of partly new 8- -methyl-9H-1 ,3-dioxolo[4,5-h][2,3]benzodiazepine derivatives having antiaggressive activity is known from HU-P No. 191 702. According to the novel process, the suitably substituted 2-acetonyl-4,5-methylenedioxybenzophenone is reacted with an excess of hydrazine hydrate.

Further 7,8-dihydro-8-methyl-9H-1 ,3-dioxolo[4,5- -h][2,3]benzodiazepine derivatives having antidepressant and antiparkinsonian activities are known from HU-P No. 206 719.

Some of the 2,3-benzodiazepine derivatives elicit their effect through the non-competitive inhibition of the AMPA/kainate receptors [Donevan, S.D. et al., J. Pharmacol. Exp. Ther., 271, 25-29 (1994)].

From the literature it is known that AMPA/kainate receptors play an important role in the acute and chronic diseases of the central nervous system. Through the inhibition of these receptors, muscle relaxant, neuroprotective and spasm inhibiting effects can be achieved [Vizi, E.S. et al., CNS Drug Reviews, 2, 91-126 (1996); Lees, G.L., CNS Drugs, 5, 51-74 (1996)].

Unpublished Hungarian patent applications Ser. No. 1380/97 and 1381/97 and PCT application No. HU 98/00075 relate to 8-substituted-9H-1 ,3-dioxolo[3,4-h][2,3]benzodiazepine derivatives having muscle relaxant, neuroprotective and anticonvulsive effect. Summary of the invention

It has been found that 8-substituted-9H-1 ,3-dioxolo[4,5- -h][2,3]benzodiazepine derivatives of the general Formula

pharmaceutically acceptable acid addition salts and quaternary ammonium derivatives thereof exhibit lipid peroxidation inhibiting effect. In the general Formula I

X represents a carbonyl group or a methylene group, and R stands for a hydrogen atom, a hydroxy group, a C₁-₄ alkoxy group, a C alkanoyloxy group, a (C1-4 alkyl)sulfonyloxy group or a group of the Formula -NR⁴R⁵, wherein

R⁴ and R⁵ mean, independently, a hydrogen atom, a C alkoxy group, a C₁-₄ alkanoyl group or a Cι-₆ alkyl group which latter is optionally substituted by a saturated or unsaturated heterocyclic group having 5 or 6 members and comprising one or two nitrogen atom(s) or a nitrogen atom and an oxygen atom as the heteroatom, or by an N-[phenyl-(Cι-₄ alkyl)]-N- -(C_M alkyl)amino group, wherein the phenyl group is optionally substituted by 1 to 3 substituent(s), wherein the substituent consists of a C alkoxy group, or

R⁴ and R⁵ form with the adjacent nitrogen atom and optionally with a further nitrogen atom or an oxygen atom a saturated or unsaturated heterocyclic group having 5 to 10 members, or

X forms together with R¹ a cyano group, a tetrazolyl group, a group of the Formula -CHNOH, or a group of the Formula -COR⁶, wherein

R⁶ means a hydroxy group, a CM alkoxy group, a phenoxy group, a naphthyloxy group, or an amino group which latter is optionally substituted by a CM alkyl group,

R² stands for a nitro group, an amino group, a hydroxy amino group (-NHOH) or a (CM alkanolyl)amino group, R³ represents a hydrogen atom, a CM alkyl group, or a group of the Formula -COR⁷, wherein

R⁷ represents a hydrogen atom, a Cι-₆ alkyl group, a Cι-₆ alkyl group substituted by 1 to 3 halo atom(s), CM alkoxy group, phenoxy group, a pyridyl group, a phenyl group or a naphthyl group which two latter groups may be optionally substituted by 1 to 3 substituent(s), or a group of the Formula -(CH₂)_n- -NR⁸R⁹, wherein

R⁸ and R⁹ represent, independently, a hydrogen atom, a CM alkyl group optionally substituted by a phenyl group or a saturated heterocyclic group having 5 or 6 members and containing a nitrogen group or a nitrogen and an oxygen group, and said phenyl group may be optionally substituted by 1 to 3 substituent(s), wherein the substituent consists of a C_M alkoxy group, or

R⁸ and R⁹ form, together with the adjacent nitrogen atom and optionally a further nitrogen or oxygen atom, a saturated or unsaturated heterocyclic group having 5 or 6 members and being optionally substituted by a phenyl group that is optionally substituted by 1 to 3 substituents, wherein the substituent consists of a halo atom or a C_M alkoxy group, n has a value of 0, 1 or 2,

Y is a hydrogen atom, or a methyl group, or

Y forms together with R³ a valence bond between the carbon atom in position 8 and the nitrogen atom in position 7.

The basis of our invention is the discovery that compounds of the general Formula I, their therapeutically useful acid addition salts and quaternary ammonium derivatives possess lipid peroxidation inhibitory effect. Detailed description of the invention

These compounds are excellent antioxidants which, among others, can be used in the following therapeutic indications. Atherosclerosis, caused by increased peroxidation of blood lipids, and ischemia of different organs due to atherosclerosis, like ischemic heart disease, cerebral ischemia and renal failure can all be prevented by long term treatment with compounds of the general Formula I. Outstanding therapeutic effect can be expected if the above diseases are accompanied by elevated plasma cholesterol or triglyceride concentration or in diabetic patients. Further use of compounds of Formula I is cytoprotection in conditions characterized by excessive production of free radicals i.e. protection against reperfusion damage caused by restoration of blood flow after thrombolysis in myocardial or cerebral infarction. In case of ischemic heart disease an antiarrhythmic effect can also be expected. Scavenging free radicals can also be used for reducing or even fully preventing symptoms and organ damage in several other diseases in which increased production of or diminished natural protection against free radicals are pathogenic factors, like in case of acute pancreatitis, rheumatoid arthritis, cataract, inflammatory diseases (e.g. hepatisis), Parkinson's disease etc. Inhibition of lipid peroxidation by long term drug therapy can provide protection against free radicals liberated during natural physiological processes thereby can slow down ageing and can reduce free radical damage of DNA. Furthermore, compounds of the general Formula I can produce beneficial therapeutic effect by scavenging free radicals produced due to radiation or chemical reactions related to drugs and toxic substances ingested. The basis of our invention is the discovery that compounds of the general Formula I, their therapeutically useful acid addition salts and quaternary ammonium derivatives can be used for treatment of states or diseases related to increased (pathological) lipid peroxidation.

These compounds can particularly be used for treatment of diseases listed below:

Diseases caused by increased production of free radicals, e.g. atherosclerosis, pancreatitis, rheumatoid arthritis, cataract, Parkinson's disease, inflammatory diseases (e.g. hepatitis), ischemic heart disease, cerebral ischemia and renal failure.

Furthermore, prevention of damage in disease states related to increased production of free radicals particularly due to radiation or drugs and toxic substances.

The definition of the terms used in the text of the patent specification and the claims is the follows:

Under a C_M alkoxy group primarily a methoxy, ethoxy, n-propoxy, isopropoxy or n-butoxy group, preferably a methoxy group is meant.

A C_M alkyl group is a methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, ter.-butyl or isobutyl group. Preferably, a C_M alkyl group is a methyl or an ethyl group.

A Ci-₆ alkyl group can be, in addition to alkyl groups listed above, for example a n-pentyl, 2-methylbutyl, n-hexyl, 2,2-dimethylbutyl or 2,3-dimethylbutyl group etc. A C alkanoyl group is, primarily, a formyl, acetyl or n- propionyl group. Preferably, a C_M alkanoyl group is an acetyl group.

Similarly, a C_M alkanoyloxy group is, primarily, a formyloxy, acetyloxy or n-propionyloxy group.

Under a saturated or unsaturated heterocyclic group having 5 or 6 members and comprising one or two nitrogen atom(s) or a nitrogen atom and an oxygen atom as the heteroatom, for example a pyrrolidinyl, piperidinyl, piperazinyl, imidazolyl or morpholino group is meant. Suitably, the other nitrogen atom of the piperazinyl group is substituted.

When the substituents R⁴ and R⁵ form with the adjacent nitrogen atom a saturated or unsaturated heterocyclic group having 5 to 10 members, said heterocyclic group contains one or two nitrogen atom(s) or a nitrogen atom and an oxygen atom as the heteroatom, and it consists of one ring or two condensed rings. The heterocyclic ring(s) contain(s) no double bond or one or more double bond(s). The above heterocyclic group is for example a pyrrolidinyl, imidazolyl, piperidinyl, pyridyl, morpholino, piperazinyl or 1,5-diazabicyclo[4.3.0]non- -5-enyl group. Suitably, one of the nitrogen atoms of the piperazinyl group is substituted.

Under a pharmaceutically suitable acid addition salt an acid addition salt formed with a pharmaceutically suitable inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid etc. or with a pharmaceutically suitable organic acid such as formic acid, acetic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, succinic acid, citric acid, methanesulfonic acid etc. is meant.

A quaternary ammonium derivative is a derivative wherein one of the nitrogen atoms of a compound of the Formula I is present in a quaternerized form.

The invention includes any isomers of the compounds of the Formula I and the mixtures thereof.

Under the isomers of the compounds of the Formula I - due to the presence of at least one chiral centre both enantiomers, and - because of isomerisms that exist in case of certain substitutions - the isomers E and Z, diastereomers, tautomeric forms, and the mixtures thereof such as the racemate are meant.

Compounds of the general Formula I, wherein Y stands for methyl, possess preferable lipid peroxidation inhibiting properties.

Compounds of the general Formula I, wherein X and R¹ together form a cyano group, also exhibit advantageous lipid peroxidation inhibiting effect.

A group of the compounds of the general Formula I exhibiting preferable lipid peroxidation inhibiting effect consists of derivatives in which R³ is a group of the Formula -COR⁷ (wherein R⁷ represents Cι-₆ alkyl, preferably methyl) or a group of the Formula -(CH₂)_n-NR⁸R⁹ (wherein R⁸ and R⁹ together with the nitrogen atom they are attached to, form a piperazino ring substituted by a phenyl group which is substituted by fluorine, chlorine or Ci-₆-alkoxy, particularly methoxy group).

The following compound of the general Formula I possesses particularly strong lipid peroxidation inhibiting properties:

7-acetyl-5-(4-aminophenyl)-7,8-dihydro-8-cyano-8- -methyl-9H-1 ,3-dioxolo[4,5-h][2,3]benzodiazepine.

The following compounds of the general Formula I also exhibit preferably lipid peroxidation inhibiting effect:

8-cyano-7,8-dihydro-7-{3-[4-(2-fluorophenyl)-piperazinyl]- -propionyl}-8-methyl-5-(4-nitrophenyl)-9H-1 ,3-dioxolo[4,5- -h][2,3]benzodiazepine;

8-cyano-7,8-dihydro-8-methyl-7-{3-[4-(2-methoxyphenyl)- -piperazinyl]-propionyl}-5-(4-nitrophenyl)-9H-1,3-dioxolo[4,5- -h][2,3]benzodiazepine;

8-cyano-7,8-dihydro-7-{3-[4-(2-methoxyphenyl)-piperazinyl]- -propionyl}-5-(4-hydroxyaminophenyl)-9H-1 ,3-dioxolo[4,5- -h][2,3]benzodiazepine;

8-cyano-7,8-dihydro-8-methyl-5-(4-amirιophenyl)-7- -trifluoroacetyl-9H-1,3-dioxolo[4,5-h][2,3]benzodiazepine;

1-(4-aminophenyl)-4-cyano-7,8-methylenedioxy-5H-2,3- -benzodiazepine;

5-(4-aminophenyl)-8-cyano-7,8-dihydro-7-{3-[2-(3,4- -dimethoxy)-phenyl]-N-methylethylamino-propionyl}-8-methyl- -9H-1,3-dioxolo[4,5-h][2,3]benzodiazepine; 5-(4-aminophenyl)-8-cyano-7,8-dihydro-7-{3-[4-(2- -fluorophenyl)-piperazinyl]-propionyl}-8-methyl-9H-1 ,3- -dioxolo[4,5-h][2,3]benzodiazepine.

The 8-substituted-9H-1 ,3-dioxolo[4,5-h][2,3]benzodiazepine derivatives of the invention are prepared by the following methods: a) for the preparation of 8-formyl-5-(4-nitrophenyl)-9H-

-1,3-dioxolo[4,5-h][2,3]benzodiazepine of the Formula

being within the scope of the compounds of the Formula I, 8- -methyl-5-(4-nitrophenyl)-9H-1,3-dioxolo[4,5-h][2,3]benzodiazepine is reacted with an oxidizing agent; or b) for the preparation of 5-(4-nitrophenyl)-9H-1 ,3-dioxolo[4,5- -h][2,3]benzodiazepine-8-carboxylic acid of the Formula

being within the scope of the compounds of the Formula I, 8-formyl- -5-(4-nitrophenyl)-9H-1 ,3-dioxolo[4,5-h][2,3]benzodiazepine of the Formula II is reacted with an oxidizing agent; or c) for the preparation of compounds of the Formula I, wherein R¹ is an imidazolyl group, R² represents a nitro group, X stands for a carbonyl group, and Y forms together with R³ a valence bond , 5-(4-nitrophenyl)-9H-1 , 3-dioxolo[4,5- -h][2,3]benzodiazepine-8-carboxyiic acid of the Formula III is reacted with 1 ,1'-carbonyldiimidazole; or d) for the preparation of compounds of the Formula I, wherein R¹ is a group of the Formula -NR⁴R⁵, R² represents a nitro group, X stands for a carbonyl group, Y forms together with R³ a valence bond, R⁴ and R⁵ are as defined in connection with the Formula I, 5-(4-nitrophenyl)-9H-1 ,3- -dioxolo[4,5-h][2,3]benzodiazepine-8-carboxylic acid of the

Formula III or a reactive derivative thereof of the Formula

wherein Y¹ is a leaving group, is reacted with an amine of the Formula

HN V wherein R⁴ and R⁵ are as stated above; or e) for the preparation of compounds of the Formula I, wherein R¹ is a CM alkoxy group, R² represents a nitro group, X stands for a carbonyl group, Y forms together with R³ a valence bond , 5-(4-nitrophenyl)-9H-1 ,3-dioxolo[4,5- -h][2,3]benzodiazepine-8-carboxylic acid of the Formula III is esterified with a C alkanol; or f) for the preparation of compounds of the Formula I, wherein R¹ is a (C alkyl)sulfonyloxy group, R² represents a nitro group, X stands for a methylene group, Y forms together with R³ a valence bond, 8-formyl-5-(4-nitrophenyl)-9H-1 ,3- -dioxolo[4,5-h][2,3]benzodiazepine of the Formula II is reacted with a reducing agent, and the 8-(hydroxymethyl)-5-(4-nitrophenyl)- -9H-1 ,3-dioxolo[4,5-h][2,3]benzodiazepine obtained is reacted with a (CM alkyl)sulfonyl halide; or g) for the preparation of compounds of the Formula I, wherein R¹ represents a C alkoxy group, a C alkanoyloxy group or a group of the Formula -NR⁴R⁵, R² stands for a nitro group, Y forms together with R³ a valence bond, R⁴ and R⁵ are as stated in connection with Formula I, 8-formyl-5-(4- -nitrophenyl)-9H-1 ,3-dioxolo[4,5-h][2,3]benzodiazepine of the Formula II is reacted with a reducing agent, and the 8- -(hydroxymethyi)-5-(4-nitrophenyl)-9H-1 ,3-dioxolo[4, 5- -h][2,3]benzodiazepine obtained or a reactive alkylating derivative thereof of the Formula

wherein Q stands for a leaving group, is reacted with a C alkanol, a CM alkanecarboxylic acid or a reactive acylating derivative thereof or an amine of the Formula V, wherein R⁴ and R⁵ are as stated above; or h) for the preparation of a compound of the Formula I, wherein X forms together with R¹ a group of the Formula -CHNOH, R² represents a nitro group, Y forms together with R³ a valence bond, 8-formyl-5-(4-nitrophenyl)-9H-1 ,3-dioxolo[4,5- -h][2,3]benzodiazepine of the Formula II is reacted with hydroxylamine; or i) for the preparation of a compound of the Formula I, wherein X forms together with R¹ a cyano group, R² represents a nitro group, Y forms together with R³ a valence bond, 8-(hydroxyiminomethyl)-5-(4-nitrophenyl)-9H-1 ,3- -dioxolo[4,5-h][2,3]benzodiazepine is reacted with a dehydrating agent; or j) for the preparation of a compound of the Formula I, wherein X forms together with R¹ a tetrazolyl group, R² represents a nitro group, Y forms together with R³ a valence bond , 8-cyano-5-(4-nitrophenyl)-9H-1 ,3-dioxolo[4,5- -h][2,3]benzodiazepine is reacted with an alkaline metal azide; or k) for the preparation of 7,8-dihydro compounds of the Formula VI being a narrower group of the compounds of the Formula I, wherein X represents a carbonyl group or a methylene group, and R¹ is as defined in connection with Formula I, a compound of the Formula

wherein X and R are as stated above, is reacted with a reducing agent; or

I) for the preparation of 7,8-dihydro-7-acyl derivatives of the Formula

being a narrower group of the compounds of the Formula I, wherein X represents a carbonyl group or a methylene group, R¹ is as stated in connection with Formula I, R³ stands for a C_M alkanoyl group, a 7,8-dihydro derivative of the Formula VI, wherein X and R¹ are as defined above, is reacted with a C_M alkanecarboxylic acid or a reactive acylating derivative thereof; or m) for the preparation of compounds of the Formula I, wherein R¹ is a group of the Formula -NR⁴R⁵, R² represents a nitro group, X stands for a carbonyl group or a methylene group, one of R⁴ and R⁵ represents a C_M alkanoyl group, while the other is as defined in connection with Formula I, Y means a hydrogen atom and in this case R³ stands for a C_M alkanoyl group, or Y forms together with R³ a valence bond, a compound of the Formula I, wherein R¹ is a group of the Formula -NR⁴R⁵, wherein one of R⁴ and R⁵ means a hydrogen atom, while the other is as defined above, X, R², Y and R³ are as stated above, is reacted with a C_M alkanecarboxylic acid or a reactive acylating derivative thereof; n) for the preparation of compounds of the Formula I, wherein Y represents a methyl group, -X-R¹ stands for a cyano group, R³ is a hydrogen atom, and R² means a nitro group, the compound of the Formula

is reacted with hydrogen cyanide; or o) for the preparation of compounds of the Formula I, wherein Y represents a methyl group, R³ stands for a hydrogen atom, R² means a nitro group and -X-R¹ represents a group of the Formula -COR⁶, wherein R⁶ is as defined in connection with the Formula I, the compound of the Formula

is hydrolyzed with a mineral acid, and the carboxylic acid obtained is optionally converted to an ester or a carboxylic amide; or p) for the preparation of compounds of the Formula I, wherein Y represents a methyl group, -X-R¹ stands for a cyano group or a group of the Formula -COR⁶, R² means a nitro group, R³ is a C_M alkyl group, and R⁶ is as defined in connection with the Formula I, a compound of the Formula I, wherein Y, -X-R¹ and R² are as stated above, R³ represents a hydrogen atom, is reacted with a (CM alkyl) halide; or r) for the preparation of compounds of the Formula I, wherein Y represents a methyl group, -X-R¹ stands for a cyano group or a group of the Formula -COR⁶, R² means a nitro group, R³ is a group of the Formula -COR⁷, R⁷ represents a group of the Formula -(CH₂)_n-NR⁸R⁹, R⁶, R⁸, R⁹ and n are as defined in connection with the Formula a compound of the Formula

wherein -X-R¹, R² and n are as stated above, X¹ is a leaving group, preferably a chloro atom, is reacted with an amine of the Formula HNR⁸R⁹; and, if desired, an obtained compound of the Formula I, wherein R² represents a nitro group, R¹, R³, X and Y are as defined in connection with Formula I, is transformed into a compound of the Formula I, wherein R² represents an amino group, by reduction; and, if desired, an obtained compound of the Formula I, wherein R² represents an amino group, R¹, R³, X and Y are as stated in connection with Formula I, is reacted with a C alkanecarboxylic acid or a reactive acylating derivative thereof; and, if desired, an obtained compound of the Formula I, wherein R² represents nitro, is transformed into a compound of the general Formula I, wherein R² represents a -NHOH group, by reduction; and, if desired, an obtained base of the Formula I is converted to a pharmaceutically suitable acid addition salt or liberated from the acid addition salt; and, if desired, an obtained compound of the Formula I or pharmaceutically suitable acid addition salt thereof is converted to a quaternary ammonium derivative.

In process a) of the invention, the reaction is performed in a manner known in itself in the preparation of aldehydes (Houben-Weyl: Methoden der Organischen Chemie, Aldehyde, Band E3, Georg Thieme Veriag, Stuttgart, 1983).

A preferred oxidizing agent is selenium(IV) oxide.

In process b) of the invention, the reaction is conducted in a manner known in itself in the preparation from carboxylic acids from aldehydes (Houben-Weyl: Methoden der Organischen Chemie, Carbonsaure-Derivate, Band E5, Georg Thieme Veriag, Stuttgart, 1985; Saul Patai: The chemistry of acid derivatives, John Wiley and Sons, New York).

In processes c), d) and e) of the invention, the reactions are carried out in a manner known in itself in the transformations of carboxylic acids (Houben-Weyl: Methoden der Organischen Chemie, Carbonsaure und Carbonsaure- Derivate, Band E5, Georg Thieme Veriag, Stuttgart, 1985).

In processes f) and g) of the invention, the reactions are performed in a manner known in itself in the transformation of oxo compounds to alcohols (Houben-Weyl: Methoden der Organischen Chemie, Alkohoie, Band VI, Georg Thieme Veriag, Stuttgart, 1979). The hydroxy compound formed is reacted also in a manner known in itself with an alkylsulfonyl halide, preferably methylsulfonyl chloride in case of process f); in case of process g), the alkylsulfonyl ester of the hydroxy compound is reacted with an amine or the hydroxy compound is acylated for example with the corresponding alkanecarboxylic anhydride.

In processes h), i) and j) of the invention, the reactions are carried out in a manner known in itself in the transformations of oxo compounds (Houben-Weyl: Methoden der Organischen Chemie, Carbonsaure und Carbonsaure- Derivate, Band E5, Georg Thieme Veriag, Stuttgart, 1985; Houben-Weyl: Methoden der Organischen Chemie, Heterane, Band III, part 4, Georg Thieme Veriag, Stuttgart, 1994).

In process k) of the invention, the reduction is performed in a manner known in itself (Houben-Weyl: Methoden der Organischen Chemie, Band IV, Reduction, Georg Thieme Veriag, Stuttgart, 1989).

In processes f), g) and k) of the invention, the reducing agent is preferably sodium tetrahydroborate. It is to be noted that in case of reducing a compound of the Formula I, wherein X represents a carbonyl group, Y forms together with R³ a valence bond, R² stands for a nitro group, using an equimolar amount of sodium tetrahydroborate, only the carbonyl group is reduced. In the presence of a large excess of sodium tetrahydroborate, in addition to the reduction of the carbonyl group, the double bond between the ring nitrogen in position 7 and the ring carbon atom in position 8 becomes saturated, too.

In processes I) and m) of the invention, the acylation reactions are carried out, in general, using a reactive acylating derivative of the CM alkanecarboxylic acid such as acid halide, acid anhydride or an active ester, at a temperature from -20 to +150°C preferably in the presence of an acid binding agent and/or pyridine, in the presence or absence of an organic solvent (Houben-Weyl: Methoden der Organischen Chemie, Carbonsaure und Carbonsaure-Derivate, Band E5, Georg Thieme Veriag, Stuttgart, 1985.; S. Patai: The chemistry of amides, Interscience Publishers, 1970).

In process n) of the invention, the reaction of the compound of the Formula IX and hydrogen cyanide is carried out in a manner known from the literature (Houben-Weyl: Methoden der Organischen Chemie, Band VIII, Georg Thieme Veriag, Stuttgart).

The 8-methyl-9H-1 ,3-dioxolo[4,5-h][2,3]benzodiazepine derivative of the Formula IX can be prepared by a method that is analogous with the process described in HU-P No. 191 702. In process o) of the invention, the cyano group of the compound of the Formula X can be hydrolyzed in a manner known in itself, preferably in the presence of a mineral acid (S. Patai: The chemistry of the cyano group).

In process p) of the invention, the nitrogen atom in position 8 of the compound of the Formula I can be acylated in a manner known in itself, in general, with an acid chloride, an acid anhydride or a chlorocarbonate ester, optionally in the presence of an acid binding agent, in the presence or absence of a solvent, at a temperature from -20 to +150°C.

For the preparation of carbamoyl derivatives, the acylated derivative obtained by using an active chlorocarbonate ester is reacted with an amino compound, or a compound of the Formula I, wherein R represents a hydrogen atom, is reacted directly with the corresponding isocyanate.

In process r) of the invention, compounds of the Formula I, wherein the carbon atom in position 8 is substituted by a group of the Formula -CO-(CH₂)_n-NR⁴R⁵, can be suitably prepared by reacting the corresponding compound of the Formula XI, wherein R¹, R² and n are as stated in connection with Formula I, X stands for a leaving group, preferably a chloro atom, with an amine of the Formula HNR⁴R⁵, wherein R⁴ and R⁵ are as defined in connection with Formula I. The compound of the Formula XI can be prepared by acylating a compound of the Formula I, wherein R means a hydrogen atom. The reactions given above are performed in a manner known from the art (Houben-Weyl: Methoden der Organischen Chemie, Band XI, G. Thieme Veriag, Stuttgart, 1957; S. Patai: The chemistry of amino group, Interscience Publishers, 1968).

A compound of the general Formula I obtained may be subjected to one or more subsequent modifications, if desired.

The nitro group of the compounds of the Formula I can be converted to an amino group by reduction in a manner known in itself. The reduction can be performed for example with tin (ll)chloride or in the presence of a catalyst using a hydrogen source. For example, the catalyst is Raney nickel, palladium or platinum oxide, the hydrogen source consists of, for example, gaseous hydrogen, hydrazine, hydrazine hydrate, formic acid, trialkylammonium formate or an alkali metal formate.

Compounds of the general Formula I, wherein R² represents nitro, can be transformed into the corresponding compounds of the general Formula I, wherein R² represents a -NHOH group. Reduction may be carried out in a manner known per se.

In case of compounds of the Formula I, wherein R represents an amino group, the latter group can be acylated with a CM alkanecarboxylic acid in a manner known in itself. The acylation reaction can be performed by the method described in connection with processes I) and m).

If desired, a base of the Formula I is reacted with an inorganic or organic acid to transform it into a pharmaceutically suitable acid addition salt, or the base of the Formula I is liberated from the acid addition salt using a stronger base.

A compound of the general Formula I or a pharmaceutically acceptable acid addition salt may be converted into a quaternary ammonium derivative, if desired. The reaction may be carried out in a manner known per se.

The lipid peroxidation inhibiting effect of the compounds of the general Formula I can be determined by the following methods, in which lipid peroxidation induced by an ADP-Fe^3t complex is measured in rat liver microsome by the thiobarbiturate method. Measurement of lipid peroxidation Methods and experimental protocol

Microsome fraction was prepared by differential centrifugation from female Wistar rats in a medium containing 0.15 M KCI and 0.05 M TRIS-HCL (pH = 7.4). Incubation was performed in an 0.1 M TRIS-HCI buffer at pH = 7.4 containing 2 mM ADP, 50 μM FeSo₄ and 0.8-1.0 mg/ml microsomal protein. Final volume of the incubation medium was 1 ml. Test substances were preincubated for 10 min at 37°C then the chemical reaction was started by the addition of 0.5 mM NADPH. Incubation lasted for 20 min while the medium was shaked intensively [J.A. Buege, S.D. Aust: Methods in Enzymology 32, 302 (1978)]. The chemical reaction was stopped by addition of 2 ml thiobarbituric acid reagent (TBA). Light transmission was measured at 535 nm and the amount of TBA reactive substance was calculated by using the 1.56 x 10^"5 M^"1 cm^"1 extinction coefficient. The effect of test substance is expressed as percentage inhibition.

The results obtained are summarized in the following Table I:

Table I Lipid peroxidation inhibition

Test compound Inhibition (%) 10^~5 M

A 92 B 58 C 77 D 100 E 97 F 90 G 97 H 97

The following test compounds are used: A = (±)-7-acetyl-5-(4-aminophenyl)-7,8-dihydro-8-cyano-8-

-methyl-9H-1 ,3-dioxolo[4,5-h][2,3]benzodiazepine; B = (±)-8-cyano-7,8-dihydro-7-{3-[4-(2-fluorophenyl)-

-piperazinyl]-propionyl}-8-methyl-5-(4-nitrophenyl)-9H-

-1 ,3-dioxolo[4,5-h][2,3]benzodiazepine; C = (±)-8-cyano-7,8-dihydro-8-methyl-7-{3-[4-(2-methoxyphenyl)-

-piperazinyl]-propionyl}-5-(4-nitrophenyl)-9H-1 ,3-

-dioxolo[4,5-h][2,3]benzodiazepine; D = (±)-8-cyano-7,8-dihydro-8-methyl-7-{3-[4-(2-methoxyphenyl)-

-piperazinyl]-propionyl}-5-(4-hydroxyaminophenyl)-9H-

-1 ,3-dioxolo[4,5-h][2,3]benzodiazepine; E = (±)-8-cyano-7,8-dihydro-8-methyl-5-(4-aminophenyl)-7-

-trifluoroacetyl-9H-1 ,3-dioxolo[4,5-h][2,3]benzodiazepine; F = 1-(4-aminophenyl)-4-cyano-7,8-methylenedioxy-5H-2,3-

-benzodiazepine; G = (±)-5-(4-aminophenyl)-8-cyano-7,8-dihydro-7-{3-[2-(3,4-

-dimethoxyphenyl)-N-methylethylamino]-propionyl}-8-

-methyl-9H-1 ,3-dioxolo[4,5-h][2,3]benzodiazepine; H = (±)-5-(4-aminophenyl)-8-cyano-7,8-dihydro-7-{3-[4-(2-

-fluorophenyl)-piperazinyl]-propionyl}-8-methyl-9H-1 ,3-

-dioxolo[4,5-h][2,3]benzodiazepine.

Numerous representatives of the 2,3-benzodiazepine derivatives of the general Formula I exhibit considerable lipid peroxidation inhibiting effect in a concentration of 10^"5 M.

Compounds of Formula I are excellent antioxidants which, among others, can be used in the following therapeutic indications. Atherosclerosis, caused by increased peroxidation of blood lipids, and ischemia of different organs due to atherosclerosis, like ischemic heart disease, cerebral ischemia and renal failure can all be prevented by long term treatment with compounds of Formula I. Outstanding therapeutic effect can be expected if the above diseases are accompanied by elevated plasma cholesterol or triglyceride concentration or in diabetic patients. Further use of compounds of Formula I is cytoprotection in conditions characterized by excessive production of free radicals i.e. protection against reperfusion damage caused by restoration of blood flow after thrombolysis in myocardial or cerebral infarction. In case of ischemic heart disease an antiarrhythmic effect can also be expected. Scavenging free radicals can also be used for reducing or even fully preventing symptoms and organ damage in several other diseases in which increased production of or diminished natural protection against free radicals are pathogenic factors, like in case of acute pancreatitis, rheumatoid arthritis, cataract, inflammatory diseases (e.g. hepatitis), Parkinson's disease etc. Inhibition of lipid peroxidation by long term drug therapy can provide protection against free radicals liberated during natural physiological processes thereby can slow down ageing and can reduce free radical damage of DNA. Furthermore, compounds of Formula I can produce beneficial therapeutic effect by inhibiting free radical production due to radiation or chemical reactions related to drugs and toxic substances ingested.

The daily dose of the compounds of the general Formula I, pharmaceutically acceptable acid addition salts and quaternary ammonium derivatives thereof may very between wide ranges, depending on the seriousness of the disease, the age, body weight and condition of the patient and the mode of administration. The daily oral dose is generally 0.1-20 mg/kg. Pharmaceutical compositions containing compounds of the general Formula I are generally administered for a longer period of time in the present indications.

Pharmaceutical compositions comprising a compound of the general Formula I, a pharmaceutically acceptable acid addition salt or a quaternary ammonium derivative thereof may be administered orally, parenterally, rectally or topically. The compositions may be finished in solid or liquid form.

The solid pharmaceutical compositions may be e.g. tablets, coated tablets, hard or soft gelatine capsules. The dosage forms may contain conventional non-toxical solid carriers e.g. mannitol, lactose, cellulose, cellulose derivatives, sodium croscarmellose, starch, magnesium stearate, sodium saccharine, talc, glucose, sacharose, magnesium carbonate etc.

Suppositories suitable for rectal administration may contain as carrier e.g. polyalkylene glycols, acetylate triglycerides etc.

Liquid compositions may be prepared by dissolving or suspending the active ingredient in a suitable carrier e.g. water, sodium chloride solution, aqueous dextrose solution, glycerol, ethanol etc. The compositions may optionally contain a minor amount of suitable non-toxical auxiliary agents e.g. wetting, dispersing, emulsifying agents, buffer materials etc. (such as sodium acetate, sorbitane monolaurate, triethanol amine etc.).

Injectable compositions (subcutaneous, intramuscular or intravenous) are suitable for paranteral administration. Injectable compositions may contain e.g. water, sodium chloride solution, dextrose solution, glycerol, ethanol etc. as carrier. The compositions may optionally contain non-toxical auxiliary agents e.g. wetting, dispersing or emulsifying agents, buffer materials etc. (e.g. sodium acetate, sorbitane monolaurate, triethanol amine oleate etc.).

Topical compositions may be finished in semisolid, liquid or solid form e.g. geles, cremes, solutions, suspensions, ointments or dusting powders.

Further details of the present invention are to be found in the Examples without limiting the scope of protection to said Examples.

Example 1

(±)-8-Cvano-7.8-dihvdro-8-methyl-5-(4-nitrophenvn-9H-1.3- -dioxolo,4,5-h]r2,3]benzodiazepine

36.0 g (111.4 mmoles) of 8-methyl-5-(4-nitrophenyl)-9H- -1 ,3-dioxolo[4,5-h][2,3]benzodiazepine and 180 cm³ of glacial acetic acid are introduced into an acid-resistant steel bomb tube of 400 cm³ capacity. To the suspension, 21.75 g (334.1 mmoles) of potassium cyanide are added at a temperature of 20 to 26°C under cooling with ice-water in 20 minutes. The bomb tube is sealed and stirred at 70°C for 22 hours. After cooling, the reaction mixture is stirred with 600 cm³ of dichloromethane and 600 cm³ of water, the phases are separated, the aqueous layer is further extracted twice using 300 cm³ of dichloromethane each time, the combined organic phases are washed three times with 300 cm³ of water each time, dried over anhydrous magnesium sulfate, and evaporated. The residue is crystallized from 250 cm³ of ether, the crystals are filtered, and washed three times using 60 cm³ of ether each time.

Thus, 33.6 g (86.0 %) of the title compound are obtained. M.p.: 162-164°C.

Analysis: for C₁₈H₁ N₄0₄ (350.34) calculated: N 15.99 %; found: N 15.62 %.

¹H NMR (CDCI₃): δ 8.23 (2H, d, J=8.9 Hz), 7.78 (2H, d, J=8.9 Hz), 6.84 (1 H, s), 6.52 (1 H, s), 6.05 (1 H, d, J=1.3 Hz), 6.03 (1 H, d, J=1.3 Hz), 5.58 (1 H, s), 3.12 (1 H, d, J=14.1 Hz), 2.83 (1 H, d, J=14.1 Hz), 1.68 (3H, s). Example 2

(±)-7-Acetyl-8-cvano-7.8-dihvdro-8-methyl-5-(4-nitrophenvπ- -9H-1.3-dioxolo.4,5-h],2,31benzodiazepine

10.51 g (30 mmoles) of the compound prepared according to Example 1 are added to 44 cm³ of acetyl chloride, and the reaction mixture is stirred at 10°C for an hour. The reaction mixture is allowed to warm to room temperature, and stirred at 25°C for further 3 days, then evaporated under reduced pressure. To the evaporation residue, 250 cm³ of water are added, and the mixture is stirred for half an hour under cooling with ice-water. The crystals obtained are filtered, washed three times using 20 cm³ of cold water each time, and dried under a lamp emitting infra red radiation. 11.2 g (95.1 %) of the crude product obtained are suspended in 20 cm³ of ethanol, stirred for half an hour, then filtered. The crystals are washed twice with 10 cm³ of ethanol each time, and once with 25 cm³ of ether. After drying, 9.5 g (80.7 %) of the title compound are obtained, m.p.: 289-292°C. Analysis: for C₂₀H₁₆N₄O₅ (392.37) calculated: C 61.22 %, H 4.11 %, N 14.28 %; found: C 60.85 %, H 4.18 %, N 13.98 %.

¹H NMR (CDCI₃): δ 8.29 (2H, d, J=9.0 Hz), 7.83 (2H, d, J=9.0 Hz), 6.99 (1 H, s), 6.51 (1 H, s), 6.10 (1 H, d, J=1.2 Hz), 6.07 (1 H, d, J=1.2 Hz), 3.11 (2H, m), 2.30 (3H, s), 1.84 (3H, s). Example 3

, ±_-7-Acetyl-7,8-dihvdro-8-methyl-5-,4-nitrophenyl)-9H- -1 ,3-dioxolof4,5-h1,2,31benzodiazepine-8-carboxamide

9.8 g (24.98 mmoles) of the compound prepared according to Example 2 are added to 130 cm³ of concentrated hydrochloric acid. The reaction mixture is stirred at 5 to 10°C for 2 hours, then at 25°C for an hour, and evaporated under reduced pressure. To the evaporation residue, 120 cm³ of ethanol are added, and the solution is evaporated again. To the evaporation residue, 150 cm³ of water are added. After 30 minutes' stirring, the crystals are filtered, washed three times with 10 cm³ of water each time, twice with diisopropyl ether, and dried under a lamp emitting infra red radiation. 9.4 g (91.7 %) of the crude product obtained are transferred to a silica gel column that is eluted with ethyl acetate. The adequate fraction is evaporated, the evaporation residue is rubbed with ether, the crystals obtained are filtered, and washed with ether.

Thus, 4.5 g (43.9 %) of the title compound are obtained. M.p.: 183-184.5°C. Analysis: for C₂₀H_1δN₄O₆ (410.39) calculated: C 58.53 %, H 4.42 %, N 13.65 %; found: C 58.70 %, H 4.52 %, N 13.21 %.

¹H NMR (DMSO-d₆): δ 8.30 (2H, d, J=8.8 Hz), 7.79 (2H, d, J=8.8 Hz), 7.03 (1 H, s), 6.89 (2H, bs), 6.56 (1 H, s), 6.11 (1 H, s), 3.09 (1 H, d, J=14.2 Hz), 2.83 (1 H, d, J=14.2 Hz), 2.27 (3H, s), 1.43 (3H,s). Example 4

(±)-8-Cvano-7,8-dihvdro-8-methyl-5-(4-nitrophenyl)-7- -trifluoroacetyl-9H-1 ,3-dioxolof4,5-h1f2,31benzodiazepine

8.76 g (25 mmoles) of the compound prepared according to Example 1 are dissolved in 60 cm³ of chloroform. To the solution, 6.5 cm³ (46 mmoles) of trifluoroacetic anhydride are added, drop by drop, under cooling with icewater at 5 to 10°C in 10 minutes. The mixture is stirred at 10°C for 2 hours, at 25°C for 25 hours, then poured into 300 cm³ of ice-water. The layers are separated, the aqueous phase is extracted twice using 100 cm³ of chloroform each time. The organic phase is dried, then evaporated. The evaporation residue is crystallized in 70 cm³ of ether. After 60 minutes' stirring, the crystals are filtered, and washed three times using 100 cm³ of ether each time.

Thus, 8.6 g (77.1 %) of the title compound are obtained. M.p.: 231-234°C.

Analysis: for C₂oH₁₃F₃N₄θ5 (446.33) calculated: C 53.82 %, H 2.94 %, N 12.55 %; found: C 54.09 %, H 2.94 %, N 12.32 %.

¹H NMR (DMSO-de): δ 8.38 (2H, d, J=8.6 Hz), 7.86 (2H, d, J=8.6 Hz), 7.28 (1 H, s), 6.76 (1 H, s), 6.19 (2H), 3.44 (2H, m), 1.89 (3H, s). Example 5

(±)-8-Cvano-7.8-dihvdro-7-{2-f2-(3,4-dimethoχyphenyl)-N- -methylethylamino1-acetylj-8-methyl-5-,4-nitrophenvQ-9H-1 ,3- -dioxolof4,5-h][2.3]benzodiazepine

To 11.35 g (26.6 mmoles) of the compound prepared according to Example 16, 130 cm³ of acetonitrile and 10.4 g (53.3 mmoles) of 2-(3,4-dimethoxyphenyl)-N-methylethylamine are added. The reaction mixture is boiled for 5.5 hours, then evaporated. The residue is stirred in 100 cm³ of water at 25°C for 3 hours, then the crystals are filtered, and washed with water.

Thus, 15.2 g (97.6 %) of the title compound are obtained. M.p.: 138-140°C.

¹H NMR (CDCI₃): δ 8.28 (2H, d, J=8.9 Hz), 7.79 (2H, d, J=8.9 Hz), 6.99 (1 H, s), 6.74 (3H, m), 6.48 (1 H, s), 6.09 (1 H, d, J=1.1 Hz), 6.05 (1 H, d, J=1.1 Hz), 3.86 (3H, s), 3.83 (3H, s), 3.77 (1 H, d, J=17.1 Hz), 3.55 (1 H, d, J=17.1 Hz), 3.09 (2H, s), 2.8 (4H, m), 2.54 (3H, s), 1.86 (3H, s). Example 6

(±)-8-Cvano-7.8-dihvdro-8-methyl-5-(4-nitrophenyl)-7-(3- -chloropropionyl)-9H-1 ,3-dioxolof4,5-h1f2,31benzodiazepine

15.6 g (44.5 mmoles) of the compound prepared according to Example 1 are added to 60 cm³ (616 mmoles) of 3-chloropropionyl chloride under stirring at 5 to 10°C in 15 minutes. The mixture is stirred at 5 to 10°C for an hour, at 25°C for 6 days, then poured onto 300 cm³ of crushed ice. The mixture is stirred for 100 minutes, then extracted three times using 300 cm³ of dichloromethane each time. The organic phase is washed with 100 cm³ of 5 % aqueous sodium hydroxide solution and 100 cm³ of water, dried over anhydrous magnesium sulfate, and evaporated. The evaporation residue is boiled with 150 cm³ of ethanol, cooled, and the crystals formed are filtered.

Thus, 10.7 g (54. 6 %) of the title compound are obtained. M.p.: 216-218°C.

Analysis: for C₂₁ Hι₇CIN₄0₅ (440.85) calculated: C 57.22 %, H 3.89 %, Cl 8.04 %, N 12.71 %; found: C 57.10 %, H 4.10 %, Cl 8.02 %, N 12.41 %.

¹H NMR (DMSO-de): δ 8.34 (2H, d, J=8.8 Hz), 7.87 (2H, d, J=8.8 Hz), 7.21 (1 H, s), 6.69 (1 H, s), 6.16 (1 H, s), 6.15 (1 H, s), 3.83 (2H, m), 3.50-2.90 (4H, m), 1.75 (3H, s). Example 7

(±)-8-Cvano-7,8-dihvdro-8-methyl-7-{3-,4-(2-methoxyphenyl)- -piperazinvn-propionyli-5-(4-nitrophenyl)-9H-1 ,3-dioxolof4,5- -h].2,31benzodiazepine

To 5.95 g (13.5 mmoles) of the compound prepared according to Example 6, 100 cm³ of acetonitrile and 5.1 g (26.5 mmoles) of (2-methoxyphenyl)-piperazine are added. The reaction mixture is boiled for 3 hours, cooled, filtered, the solids are washed with water and ether. The crude product is boiled in 80 cm³ of ethanol, cooled, and filtered.

Thus, 4.8 g (59.6 %) of the title compound are obtained. M.p.: 222-223.5°C. Analysis: for C₃₂H₃₂N₆0₆ (596.65) calculated: C 64.42 %, H 5.41 %, N 14.09 %; found: C 64.78 %, H 5.45 %, N 14.08 %.

¹H NMR (CDCI₃): δ 8.29 (2H, d, J=9.0 Hz), 7.86 (2H, d, J=9.0

Hz), 7.10-6.80 (5H, m), 6.51 (1 H, s), 6.09 (1H, d, J=1.2 Hz),

6.05 (1 H, d, J=1.2 Hz), 3.86 (3H, s), 3.30-2.60 (14H, m), 1.85

(3H, s).

Example 8

(±)-8-Cvano-7,8-dihvdro-7-{3-r2-(3,4-dimethoxyphenyl)-

-N-methylamino1-propionyli-8-methyl-5-(4-nitrophenyl)-9H-

-1 ,3-dioxolof4,5-hir2,31benzodiazepine

To 6.17 g (14 mmoles) of the compound prepared according to Example 6, 70 cm³ of acetonitrile and 5.48 g (28 mmoles) of 2-(3,4-dimethoxyphenyl)-N-methylethylamine are added. The reaction mixture is boiled for 5.5 hours, then evaporated. The residue is stirred in 50 cm³ of water at 25°C for 60 minutes, the crystals are filtered. The crude product filtered is heated in 100 cm³ of water to boiling, then cooled, the crystals are filtered, washed with water and petroleum ether.

Thus, 7.1 g (81.6 %) of the title compound are obtained. M.p.: 96-98°C.

Analysis: for C₃₂H₃₃N₅0₇ (599.65) calculated: N 11.68 %; found: N 11.22 %. ¹H NMR (CDC ): δ 8.27 (2H, d, J=8.8 Hz), 7.85 (2H, d, J=8.8 Hz), 6.99 (1 H, s), 7.85-7.65 (3H, m), 6.50 (1 H, s), 6.08 (1 H, s), 6.05 (1 H, s), 3.86 (3H, s), 3.85 (3H, s), 3.20-2.60 (10H, m), 2.41 (3H, s), 1.83 (3H, s). Example 9

(±)-7-f3-(N-Benzyl-2-morpholinoethylamino)-propionyll-8- -cvano-7,8-dihvdro-8-methyl-5-(4-nitrophenyl)-9H-1 ,3- -dioxolo[4,5-h][2,31benzodiazepine

To 20 g (45.3 mmoles) of the compound prepared according to Example 6, 500 cm³ of acetonitrile and 25.66 g (113 mmoles) of N-benzyl-2-morphoiinoethylamine are added. The reaction mixture is boiled for 6 hours, then allowed to stand at 25°C for 12 hours. The N-benzyl-2-morpholinoethylamine hydrochloride precipitated is filtered, and the filtrate is evaporated. The evaporation residue is stirred in 300 cm³ of water at 25°C for 18 hours, the crystals are filtered, and washed with water. The crude product is transferred to a silica gel column that is eluted with a mixture of hexane, acetone and methanol in a ratio of 1 :3:0.1. The adequate fraction is evaporated, the residue is suspended in water, and filtered.

Thus, 15.5 g (54.8 %) of the title compound are obtained. M.p.: 78-79°C.

Analysis: for CsΛNeOe (624.70) calculated: N 13.45 %; found: N 12.93 %. ¹H NMR (CDCI₃): δ 8.26 (2H, d, J=8.9 Hz), 7.75 (2H, d, J=8.9 Hz), 7.35-7.15 (5H, m), 6.99 (1 H, s), 6.40 (1 H, s), 6.10 (1 H, d, J=1.2 Hz), 6.06 (1 H, d, J=1.2 Hz), 3.70-3.59 (6H, m), 3.08 (2H, m), 2.95-2.60 (6H, m), 2.55-2.30 (6H, m), 1.83 (3H, s). Example 10

,±)-8-Cyano-7,8-dihvdro-7-{3-[4-(2-fluorophenyl)-piperazinvπ- -propionyl , -8-methyl-5-.4-nitrophenyl)-9H-1 ,3-dioxolo[4,5- -hir2,31benzodiazepine

To 5.5 g (12.48 mmoles) of the compound prepared according to Example 6, 75 cm³ of acetonitrile and 4.0 g (22.19 mmoles) of 2-fluorophenyl-piperazine are added. The reaction mixture is boiled for 7.5 hours, then allowed to stand for 12 hours. The crystals precipitated are filtered, washed with water and ether. The crude product is dissolved in 250 cm³ of toluene, and precipitated with 150 cm³ of petroleum ether, the crystals are filtered.

Thus, 3.96 g (54.3 %) of the title compound are obtained. M.p.: 191-192°C.

Analysis: for C31H29FN6O5 (584.61) calculated: N 14.38 %; found: N 14.11 %.

¹H NMR (CDCI₃): δ 8.29 (2H, d, J=8.8 Hz), 7.86 (2H, d, J=8.8 Hz), 7.20-6.80 (5H, m), 6.51 (1 H, s), 6.09 (1 H, d, J=1.2 Hz), 6.05 (1 H, d, J=1.2 Hz), 3.30-3.05 (6H, m), 3.05-2.60 (8H, m), 1.85 (3H, s). Example 11

(±)-7-Acetyl-5-(4-aminophenyP-8-cvano-7,8-dihydro-8- -methyl-9H-1 .3-dioxolof4,5-h1f2,3]benzodiazepine

18.3 g (46.6 mmoles) of the compound prepared according to Example 4 are suspended in a mixture of 370 cm³ of ethanol and 90 cm³ of water. To the suspension, 3.7 g 10 % palladium/carbon catalyst are added, then 46.7 cm³ (941.7 mmoles) of 98 % hydrazine hydrate are added in 20 minutes while the temperature of the reaction mixture reaches 40°C and the starting compound dissolves. The mixture is stirred for 2.5 hours at room temperature. During this time, the reaction mixture cools to 25°C, and the product precipitates. The catalyst is filtered, washed twice with 200 cm³ of ethanol each time, then three times using 500 cm³ of chloroform each time. The filtrate is under reduced pressure, and 300 cm³ of water are added to the crystalline residue. After 1 hour's stirring, the crystals are filtered, washed three times using 70 cm³ of water each time, and twice using 50 cm³ of ether each time. The 14.0 g (82.8 %) of the crude product are recrystallized from 420 cm³ of ethyl acetate, the crystals are filtered, washed three times with 30 cm³ of ether each time, and dried under a lamp emitting infra red radiation.

Thus, 10.5 g (62.1 %) of the title compound are obtained. M.p.: 162-164X.

Analysis: for C₂₀H₁₈N₄O₃ (362.39) calculated: C 66.28 %, H 5.01 %, N 15.46 %; found: C 66.88 %, H 5.12 %, N 14.78 %. ¹H NMR (CDCI3): δ 7.46 (2H, d, J=8.7 Hz), 6.96 (1 H, s), 6.67 (2H, d, J=8.7 Hz), 6.65 (1H, s), 6.05 (1 H, d, J=1.2 Hz), 6.01 (1 H, d, J=1.2 Hz), 4.15 (2H, bs), 3.05 (1 H, d, J=13.9 Hz), 2.92 (1 H, d, J=13.9 Hz), 2.16 (3H, s), 1.81 (3H, s). Example 12

.± -5-(4-Aminophenvn-8-cvano-7,8-dihvdro-7-{ 3-f2-(3.4- -dimethoxyphenyl)-N-methyl-ethylamino]-propionyl}-8-methyl- -9H-1 ,3-dioxolo[4,5-h1f2,31benzodiazepine

10.0 g (16.6 mmoles) of the compound prepared according to Example 8 are transferred into a mixture of 350 cm³ of methanol and 60 cm³ of water. To the mixture, 5.0 g of 10 % palladium/carbon catalyst are added, then, in 20 minutes, 30.0 cm³ (605 mmoles) of 98 % hydrazine hydrate are added at 15 to 20°C. The mixture is stirred at room temperature for 6.5 hours, the catalyst is filtered, and washed three times using 100 cm³ of methanol each time. The filtrate is evaporated under reduced pressure, and, to the residue, 100 cm³ of water are added. After 1 hour's stirring, the crystals are filtered, and washed three times with 30 cm³ of water each time. The crude product is transferred to a silica gel column that is eluted with a mixture of chloroform and methanol in a ratio of 4:1. The adequate fraction is evaporated, and the residue is stirred in 30 cm³ of ether for half an hour. The crystals obtained are filtered, and washed with ether.

Thus, 3.5 g (33.7 %) of the title compound are obtained. M.p.: 148-150°C. Analysis: for C32H₃₅N₅O₅ (569.64) calculated: N 12.29 %; found: N 11.89 %.

¹H NMR (CDCI₃): δ 7.48 (2H, d, J=8.6 Hz), 6.96 (1 H, s), 6.92-

6.64 (3H, m), 6.62 (2H, d, J=8.6 Hz), 6.62 (1H, s), 6.05 (1H, d,

J=1.3 Hz), 5.98 (1 H, d, J=1.3 Hz), 4.15 (2H, bs), 3.85 (6H, s),

3.04 (1 H, d, J=14.1 Hz), 2.92 (1 H, d, J=14.1 Hz), 2.88-2.54

(8H, m), 2.32 (3H, s), 1.80 (3H, s).

Example 13

(±)-5-(4-Aminophenyl)-8-cyano-7,8-dihvdro-7-( 3-f4-(2- fluorophenyl)-piperazinvn-propionyl}-8-methyl-9H-1 ,3-dioxolor4,5-

-h1[2,31benzodiazepine

16.4 g (27.5 mmoles) of the compound prepared according to Example 10 are transferred into 180 cm³ of ethanol. To the mixture, 7.26 g (32.2 mmoles) of crystalline tin(ll) chloride (SnCI₂.2H₂0) are added, and the reaction mixture is boiled for 3.5 hours. After cooling, the reaction mixture is evaporated under reduced pressure. To the residue, 180 cm³ of water are added. The mixture is make alkaline by the addition of 135 cm³ of 40 % aqueous sodium hydroxide solution, and extracted three times using 400 cm³ of dichloromethane each time. The dichloromethane phase is dried, and evaporated under reduced pressure. To the evaporation residue, 50 cm³ of ether are added, the mixture is stirred for 30 minutes, the crystals are filtered, and washed with ether. The crude product obtained is transferred to a silica gel column that is eluted with a mixture of chloroform and methanol in a ratio of 9:1. The adequate fraction is evaporated, and the residue is crystallized from 10 cm³ of ether. The crystals are filtered, and washed with ether.

Thus, 1.85 g (34.9 %) of the title compound are obtained. M.p.: 159-161 °C.

Analysis: for C₃₁ H₃₁ FN₆0₃ (554.63) calculated: C 67.13 %, H 5.63 %, N 15.15 %; found: C 66.50 %, H 5.50 %, N 15.11 %.

¹H NMR (CDCI₃): δ 7.50 (2H, d, J=8.8 Hz), 7.15-6.8 (4H, m), 6.96 (1 H, s), 6.67 (2H, d, J=8.8 Hz), 6.65 (1 H, s), 6.05 (1 H, d, J=1.3 Hz), 5.98 (1 H, d, J=1.3 Hz), 4.19 (2H, bs), 3.09 (4H, t, J=4.8 Hz), 3.05-2.68 (6H, m), 2.65 (4H, t, J=4.8 Hz), 1.81 (3H, s). Example 14

,±)-5-(4-Aminophenyl)-8-cvano-7,8-dihvdro-8-methyl-7- -trifluoroacetyl-9H-1₁3-dioxolor4,5-hir2,31benzodiazepine

4.0 g (8.96 mmoles) of the compound prepared according to Example 4 are transferred into 160 cm³ of ethanol, 9.0 g (40.0 mmoles) of crystalline tin(ll) chloride (SnCI₂.2H₂0) are added, and the mixture is boiled for 90 minutes. After cooling, the reaction mixture is evaporated under reduced pressure. To the residue, 120 cm³ of water are added, and the mixture is extracted twice using 150 cm³ of dichloromethane each time. The organic phase is washed twice with 30 cm³ of 5 % aqueous sodium hydroxide solution each time, then once with 100 cm³ of water. The pH of the aqueous phase is adjusted with 10 % aqueous sodium hydroxide solution to a value of 10, and it is extracted three times using 70 cm³ of dichloromethane each time. The dichloromethane layers obtained before and after the alkalization are combined, dried, and evaporated under reduced pressure. To the evaporation residue, 50 cm³ of diisopropyl ether are added, after 60 minutes' stirring, the crystals are filtered, and washed three times using 10 cm³ of diisopropyl ether each time. The 1.7 g (45.6 %) of crude product obtained are transferred to a silica gel column that is eluted with pure chloroform. The R_f value of the product amounts to 0.53 in a mixture of toluene and methanol in a ratio of 7:3. The fraction containing the product is evaporated under reduced pressure, the residue is crystallized from 10 cm³ of n-hexane. The crystals are filtered, and washed with 10 cm³ of n-hexane. Thus, 0.7 g (18.7 %) of the title compound are obtained. M.p.: 129-130°C. Analysis: for C₂oH₁₅F₃N₄θ3 (416.36) calculated: N 13.46 %; found: N 13.12 %.

¹H NMR (CDCI₃): δ 7.49 (2H, d, J=8.7 Hz), 6.98 (1 H, s), 6.67 (2H, d, J=8.7 Hz), 6.66 (1 H, s), 6.09 (1 H, d, J=1.3 Hz), 6.05 (1 H, d, J=1.3 Hz), 4.14 (2H, bs), 3.15 (1 H, d, J=14.4 Hz), 2.98 (1 H, bs), 1.89 (3H, s). Example 15

8-Cvano-5-(4-nitrophenylV9H-1.3-dioxolor4.5-hir2,3lbenzodiazepine 2.00 g (5.7 mmoles) of the oxime obtained in Example 6 are suspended in 100 cm³ of dichloromethane. To the suspension obtained, 1.37 g (1.90 cm³ , 13.6 mmoles) of triethylamine, then, 0.78 g (0.53 cm³, 6.8 mmoles) of methanesulfonyl chloride in 10 cm³ of dichloromethane are added, drop by drop, under cooling with ice-water. The reaction mixture is stirred at room temperature for 4 hours, then washed twice with 30 cm³ of water each time, once with 30 cm³ of saturated brine, dried over anhydrous magnesium sulfate, and evaporated under reduced pressure. The crude product obtained is recrystallized from 75 cm³ of acetonitrile, the crystals are washed with 20 cm³ of diethyl ether.

Thus, 1.27 g (67 %) of the title compound are obtained. M.p.: 230-234°C.

¹H NMR [CDCI₃ + (CD_3.2SO]: δ 8.30 (2H, d, J=8.6 Hz), 7.89 (2H, d, J=8.6 Hz), 6.92 (1 H, s), 6.72 (1 H, s), 6.15 (1 H, s), 6.13 (1 H, s), 3.67 (1 H, d, J=13.8 Hz), 3.17 (1H, d, J=13.8 Hz). Example 16

5-(4-aminophenyl^')-8-cvancr-9H-1,3-dioxolof4.5-hir2,31benzodiazepine 5.0 mmoles of nitro compound are dissolved in a mixture of 100 cm³ of dichloromethane and 100 cm³ of methanol, and the solution is hydrogenized in the presence of 0.10 g of 10 % palladium/carbon catalyst at room temperature and 5.065x10⁵ Pa pressure. After the hydrogenization, the catalyst is filtered, the solvent is evaporated under reduced pressure, and the crude product is recrystallized.

Thus, 5-(4-aminophenyl)-8-cyano-9H-1 ,3-dioxolo[4,5- -h][2,3]benzodiazepine is obtained. Analysis: for Cι₇Hι₂N₄0₂ (304.31) calculated: C 67.10 %, H 3.97 %, N 18.41 %; found: C 65.65 %, H 4.07 %, N 18.06 %.

¹H NMR [(CD₃)₂SO]: δ 7.32 (2H, d, J=8.6 Hz), 7.19 (1 H, s), 6.82 (1 H, s), 6.60 (2H, d, J=8.6 Hz), 6.17 (1 H, s), 6.12 (1 H, s), 5.82 (2H, broad s), 3.75 (1 H, d, J=13.9 Hz), 3.12 (1 H, d, J=13.9 Hz).

Claims

What we claim is,

1 . Use of 8-substituted-9H-1 ,3-dioxolo[4,5- -h][2,3]benzodiazepine derivatives of the general Formula

(wherein

X represents a carbonyl group or a methylene group, and

R¹ stands for a hydrogen atom, a hydroxy group, a CM alkoxy group, a CM alkanoyloxy group, a (CM alkyl)sulfonyloxy group or a group of the Formula -NR⁴R⁵, wherein

R⁴ and R⁵ mean, independently, a hydrogen atom, a CM alkoxy group, a C_M alkanoyl group or a Cι-6 alkyl group which latter may be optionally substituted by a saturated or unsaturated heterocyclic group having 5 or 6 members and comprising one or two nitrogen atom(s) or a nitrogen atom and an oxygen atom as the heteroatom, or by an N-[phenyl-(Cι. alkyl)]-N-(Cι.₄ alkyl)amino group, wherein the phenyl group may be optionally substituted by 1 to 3 substituent(s), wherein the substituent consists of a C alkoxy group, or

R⁶ means a hydroxy group, a C_M alkoxy group, a phenoxy group, a naphthyloxy group, or an amino group which latter may be optionally substituted by a C_M alkyl group,

R⁷ represents a hydrogen atom, a C^ alkyl group, a C-ι-₆ alkyl group substituted by 1 to 3 halo atom(s), CM alkoxy group, phenoxy group, a pyridyl group, a phenyl group or a naphthyl group which two latter groups may be optionally substituted by 1 to 3 substituent(s), or a group of the Formula -(CH₂)_n- -NR⁸R⁹, wherein

R⁸ and R⁹ represent, independently, a hydrogen atom, a C_M alkyl group optionally substituted by a phenyl group or a saturated heterocyclic group having 5 or 6 members and containing a nitrogen group or a nitrogen and an oxygen group, and said phenyl group may be optionally substituted by 1 to 3 substituent(s), wherein the substituent consists of a CM alkoxy group, or

R⁸ and R⁹ form, together with the adjacent nitrogen atom and optionally a further nitrogen or oxygen atom, a saturated or unsaturated heterocyclic group having 5 or 6 members and being optionally substituted by a phenyl group that may be optionally substituted by 1 to 3 substituents, wherein the substituent consists of a halo atom or a C_M alkoxy group, n has a value of 0, 1 or 2,

Y is a hydrogen atom, or a methyl group, or

Y forms together with R³ a valence bond between the carbon atom in position 8 and the nitrogen atom in position 7) and pharmaceutically acceptable acid addition salts and quaternary ammonium derivatives thereof for the preparation of pharmaceutical compositions suitable for the treatment or prophylaxis of diseases and states related to increased (pathological) lipid peroxidation.

2. Use according to Claim 1 for the preparation of pharmaceutical compositions for the treatment or prophylaxis of diseases caused by increased production of free radicals, e.g. atherosclerosis, pancreatitis, rheumatoid arthritis, cataract, Parkinson's disease, inflammatory diseases (e.g. hepatitis), ischemic heart disease, cerebral ischemia and renal failure.

3. Use according to Claim 1 for the preparation of pharmaceutical compositions for the prevention of damage in disease states related to increased production of free radicals, particularly due to radiation or drugs and toxic substances.

4. Use according to any of Claims 1-3 of compounds of the general Formula I wherein Y stands for methyl.

5. Use according to any of Claims 1-4 of compounds of the general Formula I wherein X and R¹ together form cyano.

6. Use according to any of Claims 1-5 of compounds of the general Formula I wherein R³ stands for a group of the Formula -COR⁷ and R⁷ is C-ι.₆-alkyl, particularly methyl.

7. Use according to any of Claims 1-6 of compounds of the general Formula I wherein R³ is a group of the general Formula -(CH₂)_n-NR⁸R⁹ and R⁸ and R⁹ together with the adjacent nitrogen atom, they are attached to, form a piperazine ring substituted by phenyl which bears one or more fluorine, chlorine or C^-alkoxy, particularly methoxy substituent(s).

8. Use according to any of Claims 1-7 of the following compound of the general Formula I:

7-acetyl-5-(4-aminophenyl)-7,8-dihydro-8-cyano-8- methyl-9H-1 ,3-dioxolo[4,5-h][2,3]benzodiazepine.

9. Use according to any of Claims 1-7 of the following compounds of the general Formula I:

8-cyano-7,8-dihydro-8-methyl-7-{3-[4-(2-methoxyphenyl)- -piperazinyl]-propionyl}-5-(4-nitrophenyl)-9H-1 ,3-dioxolo[4,5- -h][2,3]benzodiazepine;

8-cyano-7,8-dihydro-7-{ 3-[4-(2-methoxyphenyl)- -piperazinyl]-propionyl}-5-(4-hydroxyaminophenyl)-9H-1 ,3- -dioxolo[4,5-h][2,3]benzodiazepine;

8-cyano-7,8-dihydro-8-methyl-5-(4-aminophenyl)-7- -trifIuoroacetyl-9H-1 ,3-dioxolo[4,5-h][2,3]benzodiazepine;

1-(4-aminophenyl)-4-cyano-7,8-methylenedioxy-5H-2,3- -benzodiazepine;

5-(4-aminophenyl)-8-cyano-7,8-dihydro-7-{3-[2-(3,4- -dimethoxy)-phenyl]-N-methylethylamino-propionyl}-8-methyl- -9H-1 ,3-dioxolo[4,5-h][2,3]benzodiazepine;

5-(4-aminophenyl)-8-cyano-7,8-dihydro-7-{3-[4-(2- -fluorophenyl)-piperazinyl]-propionyl}-8-methyl-9H-1 ,3- -dioxolo[4,5-h][2,3]benzodiazepine.

10. Pharmaceutical compositions for the treatment or prophylaxis of diseases or states related to increased (pathological) lipid peroxidation comprising a compound of the general Formula I according to Claim 1 or a pharmaceutically acceptable acid addition salt or quaternary ammonium derivative thereof.

11. Pharmaceutical composition according to Claim 1 for the treatment or prophylaxis of diseases caused by increased production of free radicals, e.g. atherosclerosis, pancreatitis, rheumatoid arthritis, cataract, Parkinson's disease, inflammatory diseases (e.g. hepatitis), ischemic heart disease, cerebral ischemia and renal failure.

12. Pharmaceutical compositions according to Claim 10 for the prevention of damages in diseases and states related to increased production of free radicals, particularly due to radiation or drugs and toxic substances.

13. Pharmaceutical composition according to any of Claims 8-12 comprising as active ingredient the compound according to Claim 8.

14. Pharmaceutical composition according to any of Claims 8-12 comprising as active ingredient the compound according to Claim 9.

15. Method for the treatment or prophylaxis of diseases and states related to increased (pathological) lipid peroxidation which comprises administering to a patient in need of such treatment an effective amount of a compound of the general Formula I according to Claim 1 or a pharmaceutically acceptable acid addition salt or quaternary ammonium derivative thereof.

16. Method according to Claim 15 for the treatment of diseases caused by increased production of free radicals, e.g. atherosclerosis, pancreatitis, rheumatoid arthritis, cataract, Parkinson's disease, inflammatory diseases (e.g. hepatitis), ischemic heart disease, cerebral ischemia and renal failure.

17. Method of treatment according to Claim 15 for the prevention of damages in diseases and states related to increased production of free radicals, particularly due to radiation or drugs and toxic substances.

18. Method of treatment according to any of Claims 15- 17 which comprises administering to a patient a compound according to Claim 8.

19. Method of treatment according to any of Claims 15- 17 which comprises administering to a patient a compound according to Claim 9.