Background of the invention

In the Eur. J. Med. Chem. 1978, 12, 53-59, there are described tliree tetrahydroimidazo- [4,5,l-jk][l,4]benzodiazepines. EP-A-0,336,466, published October 11, 1989 discloses antiviral tetrahydroimidazo[l,4]benzodiazepinones. In Nature 1990, 343. 470, there -are described the same tetrahydroimidazo[l,4]benzodiazepinones and a few corresponding thiones.

Description of the invention

The present invention is concerned with tetrahydroimidazo[l,4]benzodiazepines having the formula :

the phaimaceutically acceptable acid addition salts and the stereochemically isomeric forms thereof, wherein

(a-2);

(a-4);

Alk is Cι_6alkanediyl; R^ is hydrogen, halo or Cι_4alkyl;

R^ and R8 each independendy are hydrogen, halo, C3_6cycloalkyl, trifluoro- methyl, 2,2,2-trifluoroethyl, Ci^alkyl optionally substituted with Cι_4alkyloxy;

R^ is hydrogen, halo or C1.4a.kyl; each RIO independently is hydrogen or Cι_4alkyl; or both R*0 taken together may form a Ci-6allcanediyl radical; n is 2, 3, 4, 5 or 6; R! 1 is hydrogen or C2-6 lkenyl; each Rl2 independently is hydrogen or Cj^alkyl; or both R^ taken together may form a Ci-6alkanediyl radical; m is 0, 1 or 2;

Rl3 is Cι_6alkyl, aryl, arylmethyl, C3_6cycloalkyl or (C3_.6cycloalkyl)Cι„4alkyl; R is hydrogen or Cj.galkyl;

R3 is hydrogen or Cι_6alkyl; R4 and R-- each independendy are hydrogen, C galkyl, halo, cyano, nitro, trifluoromethyl, hydroxy, Cj.6alkyloxy, amino, mono- or di(Cι_6alkyl)amino, Ci.βalkylcarbonylarnino or arylcarbonylamino; and

each aryl is phenyl optionally substituted with from 1 to 3 substituents independently selected from Cι_6alkyl, halo, hydroxy, Cj-galkyloxy, .amino, nitro and trifluoro¬ methyl; provided that when R^ and R-- are other than Ci.galkylcarbonylamino or .arylcarbonylamino, then R ¹ is other than C3-6alkenyl and (C3-6cycloalkyl)Ci-6alkyl.

The compounds of formula (I) may also exist in their tautomeric form. Said tautomeric form, .although not explicidy indicated in the above formula, is intended to be included within the scope of the present invention.

In the foregoing definitions the term halo is generic to fluoro, chloro, bromo and iodo; Cι.4alkyl defines straight and branched chain saturated hydrocarbon radicals having from 1 to 4 carbon atoms such as, for example, methyl, ethyl, propyl, 1-methyl- ethyl, butyl, 1-medιylpropyl, 2-methylpropyl, 1,1-dimethylethyl and the like; Ci-6alk l defines C\ gall y 1 radicals as defined hereinabove and the higher homologs thereof having from 5 to 6 carbon atoms; Cι_6alkanediyl defines bivalent straight or branched chain hydrocarbon radicals containing 1 to 6 carbon atoms such as, for example,

1,2-ethanediyl, 1,3-propanediyl, 1,4-butanediyl, 1,5-pentanediyl, 1,6-hexanediyl and the branched isomers thereof; C2-6alkenyl defines straight and branched hydrocarbon radicals containing one double bond and having from 2 to 6 carbon atoms such as, for example, ethenyl, 2-propenyl, 2-butenyl, 3-butenyl, 2-methyl-2-propenyl, pentenyl, hexenyl and the like; C3-6alkenyl defines straight and branched hydrocarbon radicals containing one double bond and having from 3 to 6 carbon atoms such as, for example,

2-propenyl, 2-methyl-2-propenyl, 2-butenyl, 3-butenyl, 2-methyl-2-butenyl, 3-methyl-2- butenyl, 4-pentenyl, 5-hexenyl and the like; C3_6alkynyl defines straight and branch chained hydrocarbon radicals containing a triple bond and having from 3 to 6 carbon atoms such as, for example, 2-propynyl, 2-butynyl, 3-butynyl, pentynyl, hexynyl -and the like ; C3_6cycloalkyl defines cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

Each RIO, R! 1 and R 2 in the radicals of formula (a-2) and (a-3), when being as defined hereinbefore but other than hydrogen, is meant to replace a hydrogen atom of the -(CH2)n- or the -CH- moiety in sad radicals.

Depending on the nature of the various substituents, the compounds of formula (I) may have several asymmetric carbon atoms . Unless otherwise mentioned or indicated, the chemical designation of compounds denotes the mixture of all possible stereo- chemically isomeric forms, said mixtures containing all diastereomers and enantiomers of the basic molecular structure. The absolute configuration of each chiral center may be indicated by the stereochemical descriptors R and S. Stereochemically isomeric forms of the compounds of formula (I) are obviously intended to be embraced within the scope of the invention.

Pure stereochemic ly isomeric forms of the compounds of formula (I) may be obtained by the application of art-known procedures. Diastereoisomers may be separated by physical separation methods such as selective cryst^zation and chromatographic techniques, e.g., counter current distribution, liquid chromatography and the like; and enantiomers may be separated from each other by the selective crystallization of their diastereomeric salts with optically active acids. Pure stereochemically isomeric forms may also be derived from the corresponding pure stereochemically isomeric forms of the appropriate starting materials, provided that the reaction occurs stereospecifically.

The compounds of formula (I) have basic properties and, consequently, they may be convened to their therapeutically active non-toxic acid addition salt forms by treatment with appropriate acids, such as, for example, inorganic acids, e.g. hydrochloric, hydro- bromic and the like acids, sulfuric acid, nitric acid, phosphoric acid and the like; or

organic acids, such as, for example, acetic, propanoic, hydroxyacetic, 2-hydroxy- propanoic, 2-oxopropanoic, ethanedioic, propanedioic, butanedioic, (Z)-2-butenedioic, (E)-2-butenedioic, 2-hydroxybutanedioic, 2,3-dihydroxybutanedioic, 2-hydroxy- 1,2,3- propanetricarboxylic, methanesulfonic, ethanesulfonic, benzenesulfonic, 4-methyl- benzenesulfonic, cyclohexanesulfamic, 2-hydroxybenzoic, 4-amino-2-hydroxybenzoic and the like acids. Conversely the salt form can be converted by treatment with alkali into the free base form. The term phaimaceutically acceptable acid addition salts also comprises the solvates which the compounds of formula (I) may form and said solvates are intended to be included within the scope of the present invention. Examples of such solvates are e.g. the hydrates, alcoholates and the like.

Interesting compounds within the invention are those compounds of formula (I) wherein R^ and R-- each independently are hydrogen, C galley 1, halo, cyano, nitro, trifluoromethyl, hydroxy, C^alkyloxy or Ci-6-alkylcarbonylamino.

More interesting compounds are those interesting compounds wherein R! is a - radical of formula (a-1) or (a-3); and/or R^ is hydrogen.

Particularly interesting compounds are those more interesting compounds wherein R! is a radical of formula (a-1), wherein R^ is hydrogen, R^ and R^ are Ci-4alkyl; and/or R^ and R^ are hydrogen or methyl; and/or R^ is hydrogen, Ci-6alkyl, halo or C l -6alkylcarbonylaιnino.

A first particular subgroup within the above defined groups comprises these compounds wherein X represents O.

A second p-articular subgroup within the above defined groups comprises these compounds wherein X represents S.

Particular compounds within the above-mentioned subgroups are these compounds wherein R* represents a radical or formula (a-1) wherein R? and R^ each independently are C3-6cyclo.alkyl, trifluoromethyl or Cl-4alkyl; or R ¹ represents a radical of formula (a-3) wherein n is 2 or 3; and the carbon atom bearing R ² has the (S)-confιguration.

More paιticul.ar compounds are those particular compounds wherein R^ .and R& each independentiy represent Ci-3alkyl; and/or each R ¹ * and R ¹² represent hydrogen. The most interesting compound is (+)-(S)-8-chloro-6-(3-ethyl-2-pentenyl)-4,5,6,7- tetrahydro-5-methyIimidazo[4,5, 1 -jk] [ 1 ,4] benzodiazepin-2(l H)-thione.

The compounds of formula (I) can generally be prepared by condensing a 9-amino- 2,3,4,5-tetrahydro-lH-l,4-benzodiazepine of formula (II) with a reagent of formula (HI), wherein L is an appropriate leaving group, such as, for example, halo, e.g. chloro or bromo.

(TO (HI)

CD

Appropriate agents of formula (III) are for example urea, di(Cι_6alkyl)carbonate, carbonoic dichloride, trichloromethyl chloroformate, l,r-carbonylbis[lH-imidazole], alkali metal, alkaline earth metal or ammonium isocy.anates, phenyl isocyanate, benzoyl isocyanate, thiourea, carbonothioic dichloride, carbon disulfide, l,r-carbonothioyl- . bis[lH-imidazole], xanthogenates, alkali metal, alkaline earth metal or ammonium isothiocyanates, phenyl isothiocyanate, benzoyl isothiocyanate, l,3-dithiolane-2-thione and the like. Said condensation reaction may conveniently be conducted by stirring and optionally heating the react^ts in a reaction-inert solvent, such as, for example, an aromatic hydrocarbon, e.g. benzene, methylbenzene, dimethylbenzene and the like; a halogenated hydrocai-bon, e.g. trichloromethane, tetrachloromethane, chlorobenzene and the like; an ether, e.g. tetrώydrofuran, 1,4-dioxane, l,l'-oxybisbutane, l,l'-oxybis- (2-methoxyethane), l,2-bis(2-methoxyethoxy)ethane and the like; a dipolar aprotic solvent, e.g. N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, l-methyl-2-pyrrolidinone, pyridine, methylpyridine, dimethylpyridine, tetrahydro- thiophene 1,1 -dioxide and the like; or a mixture of such solvents. In some instances however, it may be preferable to heat the reactants without a solvent Further it may be appropriate to add to the reaction mixture a base such as, for example, a tertiaiy amine, e.g. N. -diethylethanamine, K-ethyl-N-(l-methylethyl)-2-propanamine, 4-methyl- morpholine and the like amines. When said reagent of formula (III) is carbon disulfide, the reaction can also be conducted conveniently in an alkanol such as, for example, methanol, ethanol, propanol and the like, in the presence of a base such as sodium or potassium hydroxide and die like or in carbon disulfide as solvent and in the presence of a suitable base such as, for example, an alkyl magnesium halide, e.g. ethyl magnesium bromide, an alkyl lithium, e.g. butyllithium, an amine, e.g., N,N-diethylethanamine, a carbodiimide, e.g. £J,N-dicyclohexylcarbodiimide .and the like reagents. Or, alternatively

the latter reaction may also be conducted in basic solvent such as, for example, pyridine and the like, in the presence of a phosphite such as, for example, diphenylphosphite.

The compounds of formula (I) can also be prepared by reacting a 4,5,6,7-tetra- hydroiπώtezo{4,5, l-jk] [ 1 ,4] benzodiazepine derivative of formula (IV) with a reagent of formula M2X (V), wherein X is as defined hereinabove.

In formula (IV), ϊβ is a reactive leaving group such as, for example, halo, e.g. chloro, bromo. Appropriate reagents of formula M2X (V) are for example, water, urea, thiourea, an alkali metal thiosulfate, e.g. sodium thiosulfate and the like reagents. Sard reaction can conveniently be conducted by stirring and optionally heating the reactants in a reaction-inert solvent such as, for example, water, an alkanol, e.g., methanol, ethanol, 1-propanol, 2-propanol, butanol, 1,2-ethanediol and the like; or an aromatic hydro¬ carbon, e.g. benzene, methylbenzene, dimethylbenzene and d e like; a halogenated hydrocarbon, e.g. trichloromethane, tetrachloromethane, chlorobenzene and the like; an ether, e.g. tetrahydrofuran, 1,4-dioxane, l,l'-oxybisbutane, l,l'-oxybis(2-methoxy- ethane), l,2-bis(2-methoxyethoxy)ethane and die like; a dipolar aprotic solvent, e.g. N,N-dimedιylformamide, N,N-dimethylacetamide, dimethyl sulf oxide, 1-methyl- 2-pyrrolidinone, pyridine, methylpyridine, dimethylpyridine, tetrahydrothiophene 1,1 -dioxide and the like; or a mixture of such solvents. In some cases it may be appropriate to conduct said reaction in an excess of the reagent of formula (V), optionally in the presence of a reaction-inert solvent as defined above. In particular, the reaction may be conducted at an elevated temperature, more particularly the reflux temperature of the reaction mixture. Further, it may be appropriate to add to d e reaction mixture a base such as, for example, an amine, e.g. N,N-diethylethanamine, N-ethyl-N-(l-methyl- ethyl)-2-propanamine, 4-methylmorpholine and the like amines.

The compounds of formula (I) may also be obtained by N-alkylating an inter¬ mediate of formula (VI) with a reagent of formula R^-W (VII) wherein W represents an appropriate reactive leaving group such as, for example, halo, e.g. chloro, bromo or

iodo; or a sulfonyloxy group, e.g. benzenesulfonyloxy, 4-methylbenzenesulfonyloxy, methanesulfonyloxy and the like.

(VI) (D

Said N-alkylation reaction may conveniendy be conducted in a reaction-inert solvent such as, for example, an aromatic hydrocarbon, e.g., benzene, mediylbenzene, dimethylbenzene and the like; a lower alkanol, e.g., methanol, ethanol, 1-butanol and the like; a ketone, e.g., 2-propanone, 4-methyl-2-pentanone and the like; an ether, e.g., 1,4-dioxane, l,l'-oxybisethane, tetrahydrofuran and the like; a dipolar aprotic solvent, e.g. N,N-dimethylformamide, N,N-dimethylacetamide, nitrobenzene, dimethyl sulfoxide, l-methyl-2-pyrrolidinone, .and the like, or a mixture of such solvents. The addition of an appropriate base such as, for example, an alkali metal carbonate or hydrogen carbonate, e.g. sodium carbonate, sodium hydrogen carbonate ; sodium hydride or an organic base such as, for example, N,N-diethylethanamine or

N-(l-methylethyl)-2-propanamine and the like may be utilized to pick up die acid which is liberated during the course of the reaction. In some circumstances the addition of an iodide salt, preferably an alkali metal iodide, e.g. potassium iodide, is appropriate. Somewhat elevated temperatures and stirring may enhance the rate of the reaction.

The compounds of formula (I) wherein R* is a radical of formula (a-3) and the carbon atom of said Rl radical adjacent to the nitrogen atom bearing said R* contains at least one hydrogen atom, said radical being represented by Rl" ^a , and said compounds by formula (I-a), may also be prepared by the reductive N-alkylation of an intermediate of formula (VI) with a ketone or aldehyde of formula R l-b=O (VIII). In formula (Vlϋ), Rl" ^D represents a geminal bivalent radical derived from Rl" ³ -!! wherein two geminal hydrogen atoms are replaced by =O.

reductive

£I-alkylation

Said reductive N-alkylation reaction may conveniendy be carried out by catalytically hydrogenating the reactants in a suitable reaction-inert organic solvent according to art-known catalytic hydrogenation procedures. The reaction mixture may be stirred and/or heated in order to enhance the reaction rate. Suitable solvents are, for example, water, C^alkanols, e.g. methanol, ethanol, 2-propanol and the like; ethers, e.g. 1,4-dioxane and the like; halogenated hydrocarbons, e.g. trichloromethane .and the like; dipolar aprotic solvents, e.g. N,N-dimethylformamide, dimethyl sulfoxide and die like; esters, e.g. ethyl acetate and the like; or a mixture of such solvents. The term art- known catalytic hydrogenation procedures means that the reaction is carried out under a hydrogen atmosphere and in the presence of an appropriate catalyst such as, for example, palladium-on-charcoal, platinum-on-charcoal and the like. In order to prevent die undesired further hydrogenation of certain functional groups in the reactants and the reaction products it may be advantageous to add an appropriate catalyst-poison to the reaction mixture, e.g., thiophene and the like. Alternatively, said reductive N-alkylation may also be performed following art-known reduction procedures by treating a stirred and, if desired, heated mixture of the reactants with a reducing agent such as, for example, sodium borohydride, sodium cyanoborohydride, formic acid or a salt thereof, in particular die ammonium salt thereof.

The compounds of formula (I) wherein X is S, said compounds being represented by formula (I-b-2), can be prepared by thionation of the compounds of formula (I) wherein X is O, said compounds represented by formula (I-b-1), with 2,4-bis(4- methoxyphenyl)-l,3-didιia-2,4-diphosphetane-2,4-disulfιde (Lawesson's reagent) in an appropriate reaction-inert solvent. Such solvents are for example, aromatic hydrocar¬ bons, e.g. benzene, methylbenzene, dimethylbenzene, dipolar aprotic solvents, e.g. hexamediylphosphoric triamide (HMPA) and die like solvents.

thionation reaction

Alternatively, the compounds of formula (I-b-2) may also be obtained by thionation of the compounds of formula (I-b-1) with phosphorus pentasulfide.

The compounds of formula (I-b-2) may also be obtained by direct tiiiation of a tetrahydroimid^o[4,5,l-jk][l,4]benzodiazepine of formula (IX) with elemental sulfur at an elevated temperature.

Said reaction may conveniendy be conducted widiout a solvent at a temperature above 200°C, more particularly a temperature in the range of 230 to 250°C.

The compounds of formula (I-b-2) may alternatively be prepared by die combined reduction-thiocarbonylation of a 9-nitrobenzodiazepine of formula (X) in the presence of an alkali metal sulfide or hydrogen sulfide, and carbon disulfide.

(X)

Said reduction-thiocarbonylation reaction may conveniently be conducted by stirring the reactants in a reaction-inert solvent, optionally at an elevated temperature.

The compounds of formula (I) may also be prepared by cyclizing a benzimidazole of formula (XI) in a suitable reaction-inert solvent, optionally in the presence of a base and optionally at an elevated temperature.

CXD

In formula (XL), W represents a reactive leaving group as defined hereinbefore. Said cyclization reaction may conveniendy be conducted by stirring, and, if desired, heating the starting material. Suitable solvents are, for example, aromatic hydrocarbons, e.g. benzene, methylbenzene, dimethylbenzene -and the like, halogenated hydrocarbons, e.g. trichloromethane, tetrachloromethane, chlorobenzene and the like, ethers, e.g. tetrahydrofuran, 1,4-dioxane and the like, dipolar aprotic solvents e.g. £LM-dimethyl- formamide, M _> M-dimethylacetamide, acetonitrile, dimethylsulfoxide, pyridine and the like. Bases which may conveniendy be employed in said cyclization reaction are, for example, alkali metal or all aline earth metal carbonates, hydrogen carbonates, hydroxides, oxides, amides, hydrides and the like. In some instances the addition to the reaction mixture of a iodide salt, preferably an alkali metal iodide, e.g. potassium iodide, may be advantageous.

In all of the foregoing and in the following preparations, the reaction products may be isolated from the reaction mixture and, if necess-ary, further purified according to methodologies generally known in the art.

A number of intermediates and starting materials in the foregoing preparations are known compounds which may be prepared according to art-known methodologies of preparing said or similar compounds and some intermediates are new. A number of such preparation methods will be described herein.after in more detail.

The intermediates of formula (II) can generally be prepared from a 9-aminobenzo- diazepine of formula (Il-a) following M-alkylation reaction procedures such as described hereinabove for the preparation of the compounds of formula (I) and (I-a) from an

intermediate of formula (VI) with an alkylating reagent (VII) or with an aldehyde or ketone of formula (VIII).

(π-a)

In order to simplify the following reaction schemes, the N-alkylated intermediates wherein R* is as defined under formula (I) and die N^-unsubstituted intermediates

(wherein R is replaced by hydrogen) will be represented hereinafter by formulae wherein N is substituted with R^ _{j sa} id R H defining R ¹ and hydrogen. In intermediates (XII), (XIII), (XV), (XVI), (XVIII) and (XIX) of scheme 1 hereinbelow,

RIH also defines a radical of formula

(a-5) (a-6)

Said amide intermediates can conveniently be prepared following art-known N-acylation procedures from corresponding intermediates wherein R-Η- is hydrogen and can be reduced to the corresponding N-alkylated intermediates with complex metal hydrides or hydrides as described under reaction step A of scheme 1. In all of the following reaction schemes, the intermediates wherein RlH j _s hydrogen can also be converted into intermediates wherein RlH J _S R1 following the above described H-alkylation procedures with an alkylating reagent of formula R -W (VII) or with an aldehyde or ketone of formula R ^1_b =O (VIII).

The intermediates of formula (II-H), said intermediates representing the intermediates of formula (II) and (Il-a) can generally be prepared following the reaction steps shown in the reaction scheme 1 below.

Scheme 1

A : nitro-to-amine reduction (if RlH - _aC yi; also amide-to-amine reduction)

B : nitration

C : cyclization

D : -OH-to-W activation

E : H-alkylation : RlHNH-CH(R2)-CH(R3)OH (XXI)

The aniline derivatives in the above reaction scheme may conveniently be prepared by reduction of die corresponding nitrobenzene derivatives following art-known nitro-to- amine reduction procedures (reaction step A). Said reduction may conveniently be conducted by treatment of said nitrobenzenes with a reducing agent such as, for example, a complex metal hydride, e.g. litiiium aluminum hydride; a hydride, e.g. diborane, aluminum hydride and the like, in a reaction-inert solvent such as, for example,

l,l'-oxybisethane, tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyetiιane and the like, optionally in the presence of a cosolvent such as an aromatic hydrocarbon, e.g. benzene, methylbenzene and die like, and, if desired, at an elevated temperature. Alternatively, said reduction may also be accomplished by treatment of said nitrobenzene derivatives with sodium did ionite, sodium sulfide, sodium hydrogen sulfide, titanium(III) chloride and the like reducing agents in a suitable solvent, in particular water.

Said nitro-to-amine reduction may also be conducted following art-known catalytic hydrogenation procedures. For ex.ample, said reduction may be earned out by stiπing the reactants under a hydrogen atmosphere and in the presence of an appropriate catalyst such as, for example, palladium-on-charcoal, platinum-on-charcoal, Raney-nickel and the like catalysts. Suitable solvents are, for example, water, alkanols, e.g. ethanol, eth.anol and the like, esters, e.g. ethyl acetate and the like. In order to enhance the rate of said reduction reaction it may be advantageous to elevate the temperature and/or the pressure of the reaction mixture. Undesired further hydrogenation of certain functional groups in die reactants and die reaction products may be prevented by the addition of a catalyst poison such as, for example, thiophene and the like, to the reaction mixture.

The nitrobenzene derivatives in die above reaction scheme 1 can be prepared from benzenamine derivatives following art-known nitration procedures (reaction step B). For example, die starting materials may be nitrated by treatment with concentrated or fuming nitric acid in d e presence of concentrated sulfuric acid and optionally in the presence of a cosolvent such as, for example, a halogenated hydrocarbon, e.g. dichloromethane, trichloromethane, tetrachloromethane and the like solvents. Alternatively, said nitration may in some instances also be accomplished by adding the nitrate salt of the starting material to concentrated sulfuric acid.

The benzodiazepine derivatives (II-H), (XII) and (XIII) may be obtained from the corresponding aniline derivatives (XIV), (XV) and (XVI) (Reaction step C) following the cyclization procedures such as described hereinabove for the preparation of the compounds of formula (I) from intermediates of formula (XI). Said aniline derivatives in turn, wherein W is a reactive leaving group as defined herein¬ before, may be prepared from the corresponding alkanols by treatment with a halogena- ting reagent such as, for example, thionyl chloride, phosphoryl chloride, phosphorous trichloride and the like; or by treatment with a sulfoπylating reagent, e.g. methane- sulfonyl chloride, 4-methylbenzenesulfonyl chloride and the like (Reaction step D). Said alkanols may be prepared by N-alkylating appropriately substituted benzene derivatives of formulae (.XX), (XXII) or (XXIII) with an aminoethanol derivative of

formula R ^1H NH-CH(R ² )-CH(R ³ )OH (XXI) following art-known N-alkylation procedures such as described hereinabove (Reaction step E).

The intermediates of formula (II-H) may also be obtained following the reaction steps shown in reaction scheme 2 below. Reaction steps designated A through D are intended to refer back to the analogous reaction steps described in the previous reaction scheme.

For example, the intermediates of formula (II-H) can also be prepared from an 9-amino- or a 9-nitrobenzodiazepin-5-one of formula (XXIV) or (XXV) by reduction with a complex metal hydride e.g. lithium aluminum hydride and the like in a suitable reaction- inert solvent such as, for example, 1,2-dimethoxyethane, l,l'-oxybis(2-methoxyethane), 2,5,8,11-tetraoxadodecane, methoxybenzene and the like solvents (Reaction steps F and G). In order to enhance the rate of said reduction reaction it may be advantageous to employ an excess of the reducing reagent and to conduct said reaction at an enhanced temperature of the reaction mixture, in particular the reflux temperature of the reaction" mixture.

The intermediates of formula (XXV) can alternatively be obtained from an appropriately substituted nitrobenzene (XXXVI) by a condensation reaction (reaction step H) with a diamino reagent R ^1H NH-CH(R2)-CH(R3)-NH ₂ of formula (XXXVII) in a suitable reaction-inert solvent such as, for example, an alkanol, e.g. methanol, ethanol, 2-propanol, 1-butanol and die like; an aromatic hydrocarbon, e.g. benzene, metiiyl- benzene, dimethylbenzene and die like; a halogenated hydrocarbon, e.g. trichloro- methane, tetrachloromethane and die like; an ether, e.g. tetrahydofuran, 1,4-dioxane, l,l'-oxybisbutane, l,l'-oxybis(2-methoxyethane) and the like; a ketone, e.g. 2-propa- none, 4-medιyl-2-pentanone and die like; a dipolar aprotic solvent, e.g. N,N-dimethyl- formamide, N,N-dimethylacetamide, dimethyl sulfoxide .and the like; or a mixture of such solvents. It may be appropriate to add a base such as an alkali metal or an alkaline earth metal carbonate, e.g. sodium carbonate, sodium hydrogen carbonate and the like, to the reaction mixture. Said condensation reaction can conveniendy be conducted at an elevated temperature, in particular at the reflux temperature of the reaction mixture.

Scheme 2

(xxvπ) (XXVΠD (XXK)

(XXX) (XXXI) (xxxπ)

I I

(XXXffl) (xxxrv) (XXXV)

F : .amide-to-amine reduction

G : (nitro-to-amino) and (amide-to-amine) reduction

H : cyclization; R ^1H -NH-CH(R ² )-CH(R3)-NH ₂ (XXXVII)

I : N-acylation reaction; R - ^H NH-CH(R ² )-CH(R3)OH (XXI)

The amide derivatives (XXX), (XXXI) and (XXXII) in the above reaction scheme can conveniendy be prepared by N-acylating an ethanolamine of formula R ^1H NH-CH(R ² )-CH(R ³ )-OH (XXI) with an appropriately substituted 2-aminobenzoic acid derivative of formula (XXXIII), (XXXIV) or (XXXV) wherein L ¹ represents hydroxy or a leaving group such as, for example, halo, e.g. chloro or bromo, alkylcar- bonyloxy, e.g. acetyl, alkyloxy, e.g. methoxy, ethoxy and the like, or imidazolyl and the like leaving groups. Said N-acylation reaction (reaction step I) may be carried out by stirring the reactants in a reaction-inert solvent, optionally at an elevated temperature. In those instances where L represents hydroxy, said N-acylation reaction may also be carried out by treating the reactants with reagents capable of forming amides such as, for example, N,N-dicyclohexylcarbodiimide (DCC) option ly in die presence of a catalyst such as hydroxybenzotriazole (HOBT) or 4-dimethylaminopyridine (DMAP); 2-chloro-l- methylpyridinium iodide, l,l'-carbonylbis[lH-imidazole], l,l'-sulfonylbis[lH-imi- dazole] and the like reagents. Suitable solvents are halogenated hydrocarbons, e.g. dichloromediane, trichloromethane and the like, ethers, e.g. tetrahydrofuran, 1,4-dioxane and the like, dipolar aprotic solvents, e.g. N,N-dimethylformamide, N,N-dimethyl- acetamide, pyridine and die like; or mixtures of such solvents.

The intermediates of formula (II-H) wherein R ³ is hydrogen, said intermediates being represented by formula :

can also be prepared from a benzodiazepinedione of formula

(XXXVHO,

following the reduction procedures as described hereinabove for converting intermediates (XXrV) or (XXV) into intermediate (II-H). The preparation of the intermediates of formula (XXXVIII) may generally be conducted following the reaction padiways described in scheme 3 hereinbelow.

In all of the following reaction schemes, those compounds wherein R ³ is hydrogen, are designated by appending the suffix -α to their numerical reference.

Scheme 3

J : (nitro-to-amino) and or aliphatic amide-to-amine reduction K : cyclization to benzodiazepinedione L : N-acylation of R ^1H NH-CH(R ² )-COOR (XLVII)

In a number of the intermediates shown in scheme 3, for example in (XXXVIII), (XXXIX), (XL), (XLI), (XLϋ) and (XLIII) it is further possible to selectively reduce functional groups such as the nitro group, the ester group and/or the aliphatic amide group, in the presence of the aromatic amide group (reaction step J). Said selective reduction may be carried out by treatment of the appropriate starting material with a complex metal hydride such as, for example, lithium .aluminum hydride in a reaction-inert solvent such as, for example, tetrahydrofuran, 1,4-dioxane and the like. Alternatively, said selective reduction may also be performed by treatment of the appropriate starting material with sodium bis(2-methoxyethoxy) aluminum hydride, or with sodium borohydride in the presence of a suitable metal salt such as, for example, calcium chloride, cerium(III) chloride, aluminum chloride, zirconium(IV) chloride and die like metal salts, in a reaction-inert solvent, in particular an etiier.

The benzodiazepinediones in scheme 3 can be obtained by cyclizing (reaction step K) the corresponding acyclic intermediates of formula (XLI), (XLII) and (XLIII), wherein R represents a group such as Cι_6alkyl or aryl, a) by heating without a solvent under an inert atmosphere, optionally under reduced pressure; b) by treating with a bifunctional catalyst such as, for example, acetic acid, 2-hydroxypyridine, pyrazole, 1 ,2,4-triazole and the like, in a reaction-inert solvent such as, for example, an aromatic hydrocarbon, e.g. methylbenzene, dimethylbenzene and the like, optionally at an elevated temperature; or c) by hydrolyzing the ester and subsequendy treating the corresponding carboxylic acid (R = H) with an appropriate acid, such as, for example, a hydrohalic acid, e.g. hydrochloric acid; sulfuric acid, phosphoric acid and the like acids ; or widi a halogenating reagent such as, for example, thionyl chloride and the like.

Said intermediates in turn, can be prepared from an appropriately protected amino acid of formula R ^1H -NH-CH(R ² )-COOR (XLVII) wherein R is Ci^alkyl or aryl, by a N-acylation reaction (reaction step L) widi an appropriately substituted isatoic anhydride derivative or an appropriate 2-aminobenzoic acid derivative, by stirring the reactants at reflux temperature in .an reaction-inert solvent such as, for example, trichloromethane, pyridine and the like solvents. The intermediates of formula (II-H-α) may alternatively be prepared from benzodiazepin- 2-one derivatives following the procedures described in scheme 4.

Scheme 4

K

M M

M

M : N-alkylation of Rl ^H NH-CH(R ² )-COOR (XLVI)

The intermediates of formula (II-H) wherein R ³ is Ci.galkyl, said radical being represented by R ³ " ^a and said intermediates by formula

can be prepared by the reduction of an amine (XXIV-b) or an imine (LVII), following the reduction procedures as described hereinabove for the preparation of (II-H) from (XXIV) or (XXV).

The imine (LVII) can be prepared by reducing a nitro derivative (LVTfl) in the presence of hydrogen and a suitable metal catalyst such as, for example, palladium-on-charcoal, platinum oxide and the like catalysts. The ketone of formula (LVIII) in turn can be prepared from a 2-amino-3-nitrobenzoic acid or a functional derivative tiiereof (XXXIV) and an α-aminoketone (LIX) following art-known H-acylation procedures.

The intermediates of formula (IV) can generally be prepared from die compounds of formula (I-b-1) by reacting with a halogenating reagent such as, for example, phosphoryl chloride, phosphorous trichloride, phosphorous tribromide, thionyl chloride, oxalyl chloride and the like reagents, optionally at an elevated temperature, in particular the reflux temperature of the reaction mixture, .and optionally in the presence of a base ^' such as, for example, sodium carbonate, sodium hydrogen carbonate, potassium carbonate and die like. The reaction can be conducted in an excess of the halogenating reagent as solvent and optionally a reaction-inert solvent such as an aromatic hydrocarbon or .an etiier may be used as well.

α-b-D (IV)

The intermediates of formula (VI) can be prepared from intermediates of formula (Il-a) following the condensation reaction with a reagent of formula L-C(=X)-L (II) as described hereinbefore for the preparation of d e compounds of formula (I) from the intermediates of formula (II).

The intermediates of formula (VI) can be obtained from a benzylated compound of formula (I-c) following art-known hydrogenolysis procedures.

Hydrogenolysis

Said debenzylation reaction can be accomplished by stirring a compound of formula (I-c) in an appropriate reaction-inert solvent in the presence of a suitable metal catalyst and under a hydrogen atmosphere. Appropriate solvents are, for example, alkanols, e.g. methanol, ethanol and the like; carboxylic esters, e.g. ethyl acetate; carboxylic acids, e.g. acetic acid, propanoic acid and the like. As examples of suitable metal catalysts there may be mentioned palladium-on-charcoal, platinum-on-charcoal and the like catalysts. In order to prevent the further hydrogenation of the starting material and/or the reaction product it may be appropriate to add a catalyst-poison to the reaction mixture such as, for example thiophene.

The intermediates of formula (VI), wherein X is S, said intermediates being represented by formula (VI-b-2), may be prepared by thionation of an intermediate of formula (Vl-b- 1) following the procedures described hereinabove for the preparation of the compounds of formula (I-b-2) from (I-b-1).

thionation reaction

(Vl-b-1) (VI-b-2)

The intermediates of formula (Xfl) wherein RlH J _S hydrogen, said intermediates being represented by (Xll-a) can also be obtained by reacting an appropriately substituted nitrobenzene (LX) and a diamino reagent of formula (LXI). Herein Y is either hydrogen or a removable protective group such as, for example, Cι_6alkylcarbonyl, e.g. acetyl, trichloroacetyl and the like, a benzyl group, a C^galkyloxycarbonyl group, e.g. 1,1-dimethylethyloxycarbonyl, .and the like groups commonly used to protect .an amino group.

(LX) (LXI) (Lxπ) (Xπ-a)

Said reaction may conveniently be conducted by condensing the diamino reagent of formula (LXI) with the nitrobenzene of formula (LX), optionally removing die protective group by alkaline or acid hydrolysis or by catalytic hydrogenation and reducting die thus obtained intermediate (LXII). S d condensation reaction can conveniendy be conducted in a suitable reaction-inert solvent such as, for example, an alkanol, e.g. methanol, ethanol, 2-propanol, 1-butanol .and die like; an aromatic hydrocarbon, e.g. benzene, methylbenzene, dimethylbenzene and the like; a halogenated hydrocarbon, e.g. trichloromethane, tetrachloromediane and the like; an ether, e.g. tetrahydofuran,

1,4-dioxane, l.l'-oxybisbutane, l,l'-oxy(2-methoxyethane) and the like; a ketone, e.g. 2-propanone, 4-methyl-2-pentanone and the like; a dipolar aprotic solvent, e.g. N,N-dimethylformamide, N,N-dimethylacetamide, dimediyl sulfoxide and die like; or a mixture of such solvents. It may be appropriate to add a base such as an alkali metal or earth alkaline metal carbonate, e.g. sodium carbonate, sodium hydrogen carbonate and die like, to the reaction mixture. Said condensation reaction can conveniently be conducted at an elevated temperature, in particular at the reflux tempera-ture of the reaction mixture. Said reductions in the above procedure may conveniendy be conducted

by reacting die intermediate imines widi a suitable reductive reagent such as, for example, sodium borohydride, sodium cyanoborohydride and the like reductive reagents.

In all of die foregoing reaction schemes, the chemical designation of the intermediates defines the mixture of all possible stereochemically isomeric forms; mixtures of a number of possible stereochemically isomeric forms such as, for example, diastereomeric mixtures, enantiomeric mixtures, e.g. racemates and enriched enantiomeric mixtures; and the enantiomerically pure isomeric forms of the bωic molecular structure.

Stereochemically isomeric forms of the intermediates described in the foregoing reaction schemes and of the compounds of formula (I) may be obtained by the application of art-known procedures. For example, diastereoisomers may be separated by physical separation med ods such as destination, selective crystallization, chromatographic techniques, e.g. counter current distribution, liquid chromatography and d e like techniques.

Enantiomerically pure intermediates can conviendy be obtained from the enantiomerically pure isomeric forms of the appropriate starting materials, provided that d e subsequent reactions occur stereospecifically. Particularly interesting enantiomerically pure starting materials for use in die foregoing reaction schemes are aminoacids and/or substituted derivatives thereof, having the formula R ^1H NH-CHR ² -COOR (XL VII), and the corresponding aminoalkanols and/or substituted derivatives thereof, having the formula RlHNH-CH(R ² )-CH(R3)OH (XXI).

Alternatively, enantiomerically pure intermediates may also be obtained by separating die corresponding racemates for eχ.ample, by the selective crystallization of their diastereomeric salts with optically active resolving agents, chromatography of diastereomeric derivates, chromatography of the racemate over a chiral station^ phase and the like techniques.

The compounds of formula (I) show antiviral and in particular antiretroviral properties. Until recentiy, retroviruses were considered to be the pathogenic agents in a number of non-human warm-blooded animal diseases only, unlike viruses which have been known for quite some time to be die cause of a large number of diseases in warm- blooded animals and humans alike. However, since it has been established that a retrovirus, Human Immunodeficiency Virus (HIV), also known as LAV, HTLV-III or

ARV, is the etiological agent of Acquired Immune Deficiency Syndrome (AIDS) in

humans, retroviral infections and die treatment of subjects suffering therefrom have received the utmost attention. The HIV virus preferentially infects human T-4 cells and destroys diem or changes their normal function, particularly the coordination of die immune system. As a result, an infected patient has an everdecreasing number of T-4 cells, which moreover behave abnormally. Hence, the immunological defense system is unable to combat infections and neoplasms and the HIV infected subject usually dies by opportunistic infections such as pneumonia, or by cancers, rather than as a direct result of HIV infections. Other conditions associated with HIV infection include thrombo- cytopaenia, Kaposi's sarcoma and infection of the central nervous system characterized by progressive demyelination, resulting in dementia and symptoms such as, progressive dysarthria, ataxia and disorientation. HIV infection further has also been associated with peripheral neuropathy, progressive generalized lymphadenopadiy (PGL) and AIDS- related complex (.ARC). The antiviral, in particular antiretroviral and especially the anti- HTV properties of the compounds of formula (I) suggest said compounds to be useful antiviral chemotherapeutical agents for the prophylaxis or treatment of warm-blooded . animals suffering from viral infections, more particularly for the treatment of humans infected by HIV virus.

Due to their antiviral and in particular tiieir antiretroviral properties, d e compounds of formula (I), their pharmaceutically acceptable salts and the stereochemically isomeric forms thereof, are useful in the treatment of warm-blooded animals infected widi viruses, in particular retroviruses or for the prophylaxis of said warm-blooded animals. In general, the compounds of the present invention may be useful in the treatment of warm¬ blooded animals infected with viruses whose existence is mediated by, or depends upon, the enzyme reverse transcriptase. Examples of human retroviral infections include HIV and HTLV-I (human T-lymphotropic virus type I), causing leukemia and lymphoma. As an example of non-human animal retroviral infection tiiere may be mentioned FeLV (feline leukemia virus) which causes leukemia and immunodeficiency. Conditions which may be prevented or treated widi the compounds of the present invention, especially conditions associated wid HIV and odier pathogenic retroviruses, include AIDS, AIDS- related complex (ARC), progressive generalized lymphadenopadiy (PGL), as well as chronic CNS diseases caused by retroviruses, such as, for example HIV mediated dementia and multiple sclerosis.

In view of their antiviral, in particular antiretroviral activity, the subject compounds may be formulated into various pharmaceutical forms for administration purposes. To prepare the pharmaceutical compositions of this invention, an effective amount of the particular compound, in base or acid addition salt form, as the active ingredient is combined in intimate admixture with a pharmaceutically acceptable carrier, which cairier

may take a wide variety of forms depending on the form of preparation desired for administration.These phaπnaceutical compositions are desirably in unitary dosage form suitable, preferably, for administration orally, rectally, percutaneously, or by parenteral injection. For example, in preparing the compositions in oral dosage form, any of the usual pharmaceutical media may be employed, such as, for example, water, glycols, oils, alcohols and the like in the case of oral liquid preparations such as suspensions, syrups, elixirs and solutions: or solid carriers such as starches, sugars, kaolin, lubricants, binders, disintegrating agents and die like in the case of powders, pills, capsules and tablets. Because of their ease in administration, tablets and capsules represent the most advantageous oral dosage unit form, in which case solid pharmaceutical carriers are obviously employed. For parenteral compositions, the earner will usually comprise sterile water, at least in large part, though other ingredients, for ex-ample, to aid solubility, may be included. Injectable solutions, for example, may be prepared in which the carrier comprises saline solution, glucose solution or a mixture of saline and glucose solution. Injectable suspensions may also be prepared in which case appropriate liquid carriers, suspending agents and the like may be employed. In die compositions suitable for percutaneous administration, the carrier optionally comprises a penetration enhancing agent and/or a suitable wetting agent, optionally combined with suitable additives of any nature in minor proportions, which additives do not cause a significant deleterious effect to the skin. Said additives may facilitate the administration to the skin and/or may be helpful for preparing the desired compositions. These compositions may be administered in various ways, e.g., as a transdermal patch, as a spot-on, as an ointment. Acid addition salts of (I) due to their increased water solubility over the corresponding base form, are obviously more suitable in the preparation of aqueous compositions.lt is especially advantageous to formulate the aforementioned pharmaceutical compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used in the specification and claims herein refers to physically discrete units suitable as unitary dosages, each unit containing a predetermined quantity of active ingredient calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. Examples of such dosage unit forms are tablets (including scored or coated tablets), capsules, pills, powder packets, wafers, injectable solutions or suspensions, teaspoonfuls, tablespoonfuls and die like, and segregated multiples thereof.

The present invention is also related with a mediod of treating viral diseases in warm-blooded animals suffering from said viral diseases by administering an effective antiviral amount of a compound of formula (I), a pharmaceutically acceptable acid addition salt or a stereoisomeric form thereof. Those of skill in the treatment of viral diseases could easily determine the effective antiviral amount from the test results

presented herein. In general it is contemplated that an effective .amount would be from 0.1 mg/kg to 200 mg kg body weight, and in particular from 1 mg/kg to 50 mg/kg body weight It may be appropriate to administer the required dose as two, three, four or more sub-doses at appropriate intervals throughout the day. Said sub-doses may be formulated as unit dosage forms, for example, containing 1 to 1000 mg, and in particular 5 to 200 mg of active ingredient per unit dosage form.

The following examples are intended to illustrate and not to limit die invention in all its aspects. Unless odierwise stated, all parts therein are by weight.

Experimental part

A. Preparation of the intermediates

Example 1 a) A solution of 2.6 parts of methyl 2-bromo-3-nitrobenzoate, 1.75 parts of N-[(2-amino-l-methyl)ethyl]benzenemeth^amine and 1.06 parts of sodium carbonate in 8 parts of 1-butanol was stirred for 1/2 hour at reflux temperature. The reaction mixture was evaporated and the residue was diluted with 20 parts of water. The product was extracted with trichloromethane (2x30 parts) and d e combined extracts were dried, filtered and evaporated. The residue was converted into die hydrochloride salt. The product was filtered off, washed with 2-propanol and dried, yielding 3.4 parts (89.5%) of methyl 2-[[2-memyl-2-[(phenylmethyl)amino]ethyl]amino]-3-nitrobenzo ate hydro- chloride; mp. 204°C (interm. 1). b) A mixture of 3.8 parts of intermediate 1; 15 parts of a sodium hydroxide solution 2 N and 4 parts of 2-propanol was stirred for 1 hour at reflux temperature. While refluxing, • there was added a solution of 3 parts of concentrated hydrochloric acid and 5 parts of water. After cooling, the precipitated product was filtered off, washed widi water and recrystallized from acetic acid, yielding 3 parts (82%) of 2-[[[2-methyl-2-[(phenyl- me yl)amino]ethyl]amino]-3-nitrobenzoic acid; mp. 227°C (interm. 2). c) A mixture of 189.3 parts of intermediate 2; 400 parts of thionyl chloride and 400 parts of methylbenzene was stirred for 2 hours at reflux temperature. The reaction mixture was evaporated and the residue was taken up in 600 parts of methylbenzene. The whole was neutralized with NaHCO3 (aq.). The organic layer was separated, dried, filtered and concentrated. The residue was left at room temperature. The precipitate was filtered off, washed with 2-propanol and l,l'-oxybisethane and dried, yielding 123.5 parts of producL The mother liquor was evaporated and the residue was recrystallized from boiling 2-prop.anol. The product was filtered off at room temperature, washed with 2- propanol and l,l'-oxybisedιane and dried, yielding an additional 28 parts of product.

The combined crops were recrystallized from ethanol, yielding 137 parts (85%) of 2,3,4,5-tetrahydro-3-medιyl-9-nitro-4-(phenylmethyl)- 1H- 1 ,4-benzodiazepin-5-one; mp. 125°C (interm. 3). d) To a stirred and refluxing suspension of 14 parts of lithium aluminum hydride in 40 parts of benzene and 50 parts of tetrahydrofuran there was added a solution of 20.2 parts of intermediate 3 in 200 parts of tetrahydrofwan. Stirring at reflux temperature was continued for 2.5 houre. After cooling on ice, there were added successively water, NaOH 15% and water. The whole was filtered and die filtrate was evaporated. The residue was co-evaporated with 40 parts of methylbenzene, yielding 19.8 p.arts (87.6%) of 9-amino-2,3,4,5-tetrώydro-3-methyl-4-(phenylmethyl)-lii-l,4 -benzodiazepine (which was used without further purification in the next reaction step) (interm. 4). e) A mixture of 19.8 parts of intermediate 4 and 7.2 parts of urea was heated at 210-220°C until foaming and evolution of gaseous ammonia ceased (about 10 min). After cooling to 100°C, there were added 120 parts of HC1 1 N. The solution was decanted from the oily residue, boiled with activated charcoal and filtered. After cooling, the . filtrate was basified with ammonium hydroxide and extracted widi trichloromethane (75 and 150 parts). The combined extracts were dried, filtered and evaporated. The residue was triturated in 2-propanol and was then crystallized from ethanol and from 4-methyl- 2-pent.anone. The product was filtered off and dried, yielding 2.5 parts (11.5%) of 4,5,6,7-tetrahydro-5-med yl-6-(phenylmethyl)-imidazo[4,5,l-jk][l,4]-benzodiazepin- 2(lH)-one; mp. 205°C (interm. 5). f) A mixture of 8 parts of intermediate 5 in 80 parts of acetic acid was hydrogenated at normal pressure -and 38°C with 1 part of palladium-on-chaijcoal catalyst 10%. After the calculated amount of hydrogen was taken up, the catalyst was filtered off and d e filtrate was evaporated. The residue was taken up in 75 parts of water and the whole was basified with 30 parts of concentrated ammonium hydroxide. The mixture was left to crystallize at room temperature. The product was filtered off and recrystallized from 2-propanol, yielding 3.7 parts (66.8%) of 4,5, 6,7-teffahydro-5-methylimid.azo[4,5, l-jk] [l,4]benzodiazepin-2(lH)-one; mp. 190.5°C (interm. 6).

Example 2 a) To a stirred and cooled (-12°C) mixture of 9.10 p-aits of 2-amino-3-nitrobenzoic acid, 6.95 parts of methyl L-α-alanine monohydrochloride, 13.50 parts of 1 -hydroxy- 1H- 1,2,4-benzotriazole monohydrate and 178 parts of tetrahydrofuran there were added portionwise 5.05 parts of M-methylmorpholine and, after 5 min, 10.3 parts of

N,r^-methanetetraylbis[cyclohexanamine] under an argon atmosphere. Stirring was continued for 5 1/2 houre at -12°C and for 15 hours at room temperature. .After cooling

to 0°C for 1 2 hour, the reaction mixture was filtered and the filtrate was evaporated. The residue was partitioned between ediyl acetate and a saturated sodium hydrogen carbonate solution. The organic layer was separated, washed widi NaHCO ₃ (sat.), dried, filtered and evaporated. The residue was triturated with hexane. The product was filtered off and dried, yielding 13.08 parts (97.9%) of (-)-methyl (S)-2-[(2-amino-3-nitrobenzoyl)- aminojlpropanoate; mp. 132.9°C (interm. 7). b) A mixture of 12.58 parts of intermediate 7; 3.50 parts of palladium-on-charcoal catalyst 10% and 158 parts of ethanol was hydrogenated in a Parr apparatus for 4 hours at room temperature and a pressure of 3.1 10^ Pa. The catalyst was filtered off over diatomaceous earth and die filtrate was evaporated. The residue was placed under reduced pressure (3.3 10 ³ Pa.) and stirred at 150°C for 10 min and at 202°C for 40 min. After cooling, the solid was triturated widi edianol. The product was filtered off, washed with ethanol and l.l'-oxybisetiiane and dried, yielding 5.58 parts (57.7%) of (+)-(S)-9-amino-3,4-dihydro-3-metiιyl- 1H- 1 ,4-benzodiazepine-2,5-dione (interm. 8). c) To a suspension of 5.55 parts of lithium aluminum hydride in 154.5 parts of

1,4-dioχ.ane there were added 5.00 parts of intermediate 8 under an argon atmosphere. After refluxing for 5 hours and subsequent cooling to 10°C, there were added successively 5.55 parts of water, 9.16 parts of NaOH 15% and 16.65 parts of water. The whole was stirred for 2 hours and tiien filtered. The precipitate was washed widi 178 parts of hot tetrahydrofuran -and 133 parts of hot dichloromediane. The combined filtrates were dried, filtered and evaporated. The residue was poured into a solution of 7.36 parts of N-methylmorpholine in 133 parts of dichloromethane. The whole was added dropwise to a solution of 4.82 parts of trichloromethyl chloroformate in 160 parts of dichloromethane at 0°C under an argon stream. Stirring at 0°C was continued for 10 min. After w.aιming to room temperature, the reaction mixture was evaporated and there were added 70 parts of an aqueous 1,4-dioxane solution (15%) to the residue. The mixture was heated on a steam-bath for 45 min under a nitrogen stream, cooled and extracted wid dichloromethane (2x66.5 parts). The aqueous layer was filtered and basified widi concentrated ammonium hydroxide. The precipitate was filtered off, washed widi cold water, dried and triturated widi 2-propanol (2x), yielding 1.59 parts (32.1 %) of (+)-(S)- 4,5,6,7-tetrahydro-5-methylimidazo[4,5,l-jk][l,4]benzodiazep in-2(lH)-one; mp. 206.5°C (interm. 9).

In a similar manner there was also prepared (+)-(S)-4,5,6,7-tetrahydro-5,8-dimethyl- imidazo[4,5,l-jk][l,4]benzodiazepin-2(lH)-one; mp. 207.8°C (interm. 10). d) A mixture of 1.50 parts of intermediate 9; 1.11 parts of bromomethylcyclopropane, 1.18 parts of sodium carbonate, 1.22 parts of potassium iodide and 28.2 parts of N,N-dimethyIformamide was heated at 85°C for 24 hours under an argon atmosphere.

The reaction mixture was evaporated and the residue was partitioned between water and dichloromethane. The organic layer was separated, washed with NaOH 3N and NaCl (sat.), dried, filtered .and evaporated. To the residual oil there were added 3.95 parts of acetonitiile. After cooling at 0°C for 1 hour, the product was filtered off, washed with acetonitrile and further purified by flash column chromatography (silica gel ; CH2CI2 / C2H5OH 97:3). The eluent of the desired fraction was evaporated, yielding 0.77 parts (40.5%) of (+)-(S)-6-(cyclopropylmethyl)-4,5,6,7-tetrahydro-5-methylimi dazo- [4,5,1-jk] [l,4]benzodiazepin-2(lH)-one; mp. 115.9°C (interm. 11). e) To a stirred and cooled (-60 to -50°C) amount of 45.3 parts of concentrated nitric acid there were added portionwise 2.75 parts of intermediate 11 under an argon atmosphere.

When a clear solution was obtained, stirring and cooling was continued for 1/2 hour. The reaction mixture was slowly poured into 400 parts of ice-water and die whole was basified to pH 8 with Na2CC>3. The precipitated product was filtered off and dried in vacuo at 50°C for 16 hours, yielding 0.5 parts (15.6%) of a mixture of (S)-6-(cyclo- propylmethyl)-4,5,6,7-tetrahydro-5-methyl-9-nitroimidazo[4,5 , l-jk][l ,4]benzodiazepin- 2(lH)-one and the 8-nitro-isomer thereof (75:25) (interm. 12). f) To a refluxing mixture of 1.03 parts of hydrazine monohydrate, 23.7 parts of methanol and 0.15 parts of Raney nickel there were added portionwise 0.5 parts of intermediate 12. After refluxing for 20 min and subsequent cooling, the reaction mixture was filtered over diatomaceous earth and the filtrate was evaporated. The residue was purified by preparative thin layer chromatography (eluens : CH2CI2 / CH3OH 90:10). The eluent of the pure fractions was evaporated and the residue was dried in vacuo at 50°C for 16 hours, yielding 0.17 parts (39.0%) of (+)-(S)-9-amino-6-(cycloρropyl- methyl)-4,5 ,6,7-tetrahydro-5-methylimidazo[4,5, 1 -jk] [ 1 ,4] benzodiazepin-2( lH)-one;

25 mp. 188.7°C; [α] _D = +13.4° (cone. = 0.50% in trichloromethane) (interm. 13). g) To a refluxing mixture of 1.6 parts of Raney nickel, 15.48 parts of hydrazine monohydrate and 158 parts of methanol there were added portionwise 12.4 parts of intermediate 12. After refluxing for 20 min and subsequent cooling, the reaction mixture was filtered over diatomaceous earth and the filtrate was evaporated. The residue was triturated in acetonitrile and then taken up in trichloromethane. This solution w.as washed with water, dried, filtered and evaporated. The residue was crystallized from acetonitrile. After filtration, the mother liquor was evaporated and the residue was purified by column chromatography (silica gel ; CH ₂ C1 ₂ / CH ₃ OH(10% NH ₄ OH) 99: 1 → 97:3 → 95:5). The eluent of the desired fractions was evaporated and the residue was dried for 16 hours at 50 °C, yielding 1.25 parts (11.2%) of (+)-(S)-8-amino-6-(cyclopropylmethyl)-

4,5,6,7-tetrahydro-5-methylimidazo[4,5, l-jk] [ 1 ,4]benzodiazepin-2(lH)-one; mp.206.0 °C; [α]^ ⁵ = +4.5° (c = 0.44% in methanol) (interm. 14).

Example 3 a) A mixture of 41.49 parts of 6-chloro-lH-3,l-benzoxazine-2,4-dione and 31.40 parts of methyl L-α-alanine monohydrochloride in 108 p.arts of pyridine was refluxed for 10 hours under an argon atmosphere. After cooling, the reaction mixture was stirred for 12 hours at room temperature. The precipitate was filtered off, rinsed with water and triturated in ethanol. The product was filtered off, washed with ethanol and dried, yielding 24.77 parts (52.5%) of (S)-7-chloro-3,4-dihydro-3-methyl-lH- 1 ,4-benzo- diazepine-2,5-dione (interm. 15). b) To a cooled (0°C) amount of 142 parts of nitric acid there were added portionwise 24.55 parts of intermediate 15 under an argon atmosphere. After cooling for 3 1/2 hours at 0°C, the reaction mixture was slowly added to 450 parts of ice while stirring. The precipitate was filtered off, rinsed with water and dried at room temperature overnight, yielding 27.84 parts (93.9%) of (S)-7-chloro-3,4-dihydro-3-methyl-9-nitro-lH-l,4- benzodiazepine-2,5-dione (interm. 16). c) To a cooled (0°C) suspension of 18.2 parts of lithium aluminum hydride in 261 parts of 1,2-dimedιoxyedιane there were added portionwise 16.14 parts of intermediate 16 under a nitrogen atmosphere. The mixture was stirred for 2 hours at 0°C and for 40 hours at reflux temperature. After cooling to 0°C, there were added a mixture of 18.2 parts of water and 48.1 parts of tetrahydrofuran, 21.1 parts of NaOH 15% and 54.6 parts of water. The mixture was stirred for 1 hour at room temperature and was then filtered. The precipitate was refluxed in tetrahydrofuran for 5 min and filtered off again. The combined filtrates were dried, filtered and evaporated and die residue was dissolved in 399 parts of dichloromethane. After drying and filtering, this solution was combined with 18.2 parts of N-methylmorpholine and the whole was added dropwise to a mixture of 11.9 parts of trichloromethyl chloroformate and 665 parts of dichloromethane at 0"C and under argon. The whole was evaporated and the residue was taken up in 150 ml of a mixture of water and 1,4-dioxane 85: 15. The mixture was heated for 2 hours on a steam-bad under nitrogen. After cooling, the solid was filtered off .and dissolved in 80 parts of water. The solution was basified widi NH ₄ OH and stirred for 45 min. The product was filtered off and crystallized successively from acetonitrile and 2-propanol, yielding 2.28 parts (16%) of (+)-(S)-9-chloro-4,5,6,7-tetrahydro-5-methylimidazo[4,5, 1 -jk] [ 1 ,4] benzodiazepin-

20 2(lH)-one; mp. 202.2°C; [α]^ = +72.6° (cone. = 0.98% in methanol) (interm. 17).

In a similar manner there was also prepared 4,5,6,7-tetrahydro-5,9-dimethylimidazo- [4,5,1-jk] [l,4]benzodiazepin-2(lH)-one; mp. 199.2°C (interm. 18).

Example 4 a) To a homogeneous solution of 8.42 parts of (S)-2-aminopropanamide monohydro- bromide, 12.26 parts of sodium acetate and 79 parts of metiianol mere were added 10.96 parts of 2,6-dichloro-3-nitrobenzaldehyde and, after 1/2 hour, a mixture of 3.77 parts of sodium cyanotrihydroborate and 7.9 parts of metiianol. The whole was stirred for 45 min at room temperature. After acidifying to pH 1 widi HC1 3N, stirring was continued overnight The reaction mixture was evaporated and the residue was basified with NaHCO3 (sat.). The product was extracted widi dichloromethane and the extract was dried, filtered .and evaporated. The residue was recrystallized from 2-propanol, yielding 10.29 parts (70.7%) of (S)-2-[[(2,6-dichloro-3-nitrophenyl)methyl]amino]propanamide (interm.19). b) A mixture of 10.03 parts of intermediate (19), 348 parts of 1,2-dimethoxyethane and 92.5 parts of a solution of borane tetrahydrofurancomplex in tetrahydrofuran 1M was stirred for 3 days at room temperature under argon. There were slowly added 142 parts of methanol and 180 ml of HC1 3N and stirring was continued over weekend. The reaction mixture was basified with 200 ml of NaOH 3N and was then evaporated. The residue was extracted widi dichloromediane and the extract was dried, filtered and evaporated. The residue was refluxed in a mixture of 3.0 parts of sodium acetate and 81 parts of 1-butanol for 3 days under argon. The solvent was evaporated and the residue was dissolved in dichloromethane. This solution was washed widi NaHCO3, dried, filtered and evaporated. The residue was purified twice by flash column chromatography (silica gel ; CH2CI2 / CH3OH 99: 1). The eluent of die desired fraction was evaporated and the residue was converted into the (E)-2-butenedioate salt in methanol. The salt was filtered off (1st fraction) and the motiier liquor was evaporated. The residue was chromatographed and also converted into the salt (2nd fraction). The combined fractions were treated wid a mixture of dichloromediane and NaOH 3N to set free the base, yielding 3.73 (45.0%) of (S)-6-chloro-2,3,4,5-tetrahydro-3-methyl-9-nitro-lH-L4- benzodiazepine (interm. 20). c) To 1.89 parts of intermediate 20 under argon there were added 1.24 parts of sodium c^bonate, 1.30 parts of potassium iodide and a solution of 1.46 parts of l-bromo-3- ethyl-2-pentene in 17.86 parts of N.N-dimethylformamide. After stirring overnight, the residue was taken up in l,l'-oxybisethane. The whole was washed with water and NaCl (sat.), dried, filtered and evaporated, yielding 2.67 parts (100%) of (S)-6-chloro-4-(3-

ethyl-2-pentenyl)-2,3,4,5-tetrahydro-3-methyl-9-nitro- 1 H- 1 ,4-benzodiazepine (interm.21).

In a similar manner there was also prepared (S)-6-chloro-4-(2-cyclopentyUdeneedιyl)- 2,3,4,5-tetrahydro-3-methyl-9-nitro-lH-l,4-benzodiazepine (interm.22). d) To a cooled(0°C) mixture of 1.16 parts of lithium aluminum hydride and 26.7 parts of tetrahydrofuran under argon tiiere was added dropwise a solution of 2.57 parts of intermediate 21 in 40.05 parts of tetrahydrofuran. The whole was stirred for 1/2 hour at 0 °C, for 1 hour at room temperature and for 8 hours at reflux temperature. Then there were slowly added 1.16 parts of water, 1.16 ml of NaOH 3N, 3.48 parts of water and 35.6 parts of tetrahydrofuran. After stirring for 1/2 hour, the precipitate was filtered off and washed with hot tetrahydrofuran. The combined filtrates were evaporated and the residue was dissolved in dichloromethane. This solution was dried, filtered and evaporated, yielding 2.41 parts (100%) of (S)-6-chloro-4-(3-edιyl-2-pentenyl)-2,3,4,5- tetrahydro-3-methyl-lH-l,4-benzodiazepin-9-amine (interm. 23). e) To a refluxing mixture of 0.31 parts of Raney nickel, 2.3 parts of intermediate 22 and 71.1 p.arts of methanol under argon, there were added dropwise 2.12 parts of hydrazine monohydrate. After refluxing for 1/2 hour, the reaction mixture was evaporated and the residue was dissolved in l,l'-oxybisethane. This solution was washed with water and NaCl (sat.), dried, filtered and evaporated, yielding 2.04 parts (97.2%) of (S)-6-chloro- 4-(2-cyclopentylideneedιyl)-2,3,4,5-tetrahydro-3-methyl- IH- 1 ,4-benzodiazepin-9-amine (interm.24).

Example 5

A mixture of 4.0 parts of compound 2; 46.2 parts of phosphoryl chloride and 1.30 parts of sodium carbonate was heated at 85°C for 18 hours while nitrogen was bubbled through. The excess of phosphoryl chloride was removed and to the residue tiiere were added successively 100 parts of water, 100 ml of NaHCO3 (sat.) and 266 parts of dichloromediane. The aqueous layer was separated and re-extracted with dichloro¬ methane. The combined organic layers were dried, filtered and evaporated, yielding 1.90 parts (44.9%) of (S)-2,9-dichloro-6-(3-ethyl-2-pentenyl)-4,5,6,7-tetrahydiO-5 -methyl- imidazo[4,5,l-jk] [1,4] benzodiazepine (interm. 25).

In a similar manner compound 20 was converted into (+)-(S)-2,9-dichloro-6-(2- cyclopentylideneethyl)-4,5,6,7-tetrahydro-5-methyl-imidazo[4 ,5,l-jk][l,4]- benzodiazepine (iπterm.26).

Example 6

To a cooled (-78°C) solution of 1.23 parts of compound 18 in 93.1 parts of dichloromethane under argon, there were added successively 1.38 parts of trifluoroacetic anhydride, after 10 min, 0.79 parts of 2,6-dimethylpyridine and, after 15 min, 23.1 ml of a solution of HCl in 1 , 1 '-oxy bisethane 0.8N. The whole was left for 1 min and was dien poured into NaHCO3 (sat). The organic layer was separated, dried, filtered and evaporated, yielding 1.61 parts (100%) of (S)-2,8-dichloro-6-(3-ethyl-2-pentenyl)- 4,5,6,7-tetrahydro-5-medιylimidazo[4,5,l-jk] [1,4] benzodiazepine (interm. 27).

B. Preparation of the final compounds Example 7

To a mixture of 1.00 part of intermediate 6; 0.782 parts of sodium carbonate, 0.816 parts of potassium iodide and 94 parts of N,H-dimethylformamide there were added 1.18 parts of 2,3-dibromopropene under an argon atmosphere. The whole was heated at 65-70°C for 5 hours under argon and was dien evaporated. The residue was partitioned between dichlorometh.ane -and water. The organic layer was separated and washed with water. The combined aqueous layers were re-extracted with dichloromethane. The combined organic layers were washed widi NaCl (sat), dried, filtered and evaporated. The residue was dissolved in refluxing acetonitrile and recrystallized upon cooling (2x). The product was filtered off, washed widi cold acetonitrile and dried in vacuo at 82°C, yielding 0.762 parts (48.1%) of 6-(2-bromo-2-propenyl)-4,5,6,7-tetrahydro-5-methyl-imidazo[4 ,5,l- jk][l,4]benzodiazepin-2(lH)-one; mp. 150.0°C (comp. 1).

Example 8 To a mixture of 1.50 parts of intermediate 10; 0.77 parts of sodium carbonate and 9.4 parts of M,N-ώmemylformamide there were added 1.47 parts of l-bromo-3-ethyl-2- pentene. After heating at 60°C for 1 1/2 hour and subsequent cooling, the reaction mixture was partitioned between water and dichloromethane. The organic layer was separated, dried, filtered .and evaporated. The residue was crystallized from acetonitrile. The product was filtered off, washed with cold acetonitrile and dried, yielding 0.93 parts (43.0%) of (+)-(S)-6-(3-ethyl-2-pentenyl)-4,5,6,7-tetrahydro-5,8-dimeth ylimidazo- [4,5,l-jk][l,4]benzodiazepin-2(lH)-one; mp. 117.3°C; [α]p ⁵ = +4.9° (cone. = 1.0% in metiianol) (comp. 5).

Example 9

To a stirred solution of 0.32 parts of intermediate 13 in 26.7 parts of tetrahydrofuran there were added 0.093 parts of acetyl chloride. Stirring was continued for 16 hours at

room temperature. The reaction mixture was evaporated and the residue was basified with Na2CO3 (sat). The product was extracted widi trichloromediane and die extract was dried, filtered and evaporated. The residue was purified by flash column chromato¬ graphy (silica gel ; CH2CI2 / CH3OH 100:0 → 97:3). The eluent of the desired fraction was evaporated and die residue was dried in vacuo at 50°C for 16 hours, yielding 0.18 parts (47.7%) of (+)-(S)-N-[6-(cyclopropylmethyl)-l,2,4,5,6,7-hexahydro-5-met hyl-2-

25 oxoimidazo[4,5,l-jk][l,4]benzodiazepin-9-yl]acetamide; mp. 243.9°C; [α] _D = +15.1°

(cone. = 0.43% in methanol) (comp. 8).

Example 10

A mixture of 1.90 parts of intermediate 25; 1.50 parts of thiourea and 39.5 parts of etiianol was refluxed for 16 hours. The reaction mixture was evaporated and the residue was taken up in a mixture of 180 parts of ethyl acetate, 50 parts of water and 50 ml of NaHCO3 (sat.). The organic layer was separated, washed with water, dried, filtered and evaporated. The residue was purified by flash column chromatography (silica gel ; hexane / CH3COOC2H5 8: 1 → 4: 1 ). The eluent of the desired fraction was evaporated and die residue was converted into die hydrochloride salt in ethanol, yielding 1.74 parts (83.4%) of (-)-(S)-9-chloro-6-(3-ethyl-2-pentenyl)-4,5,6,7-tetrahydro-5 -methyl- imidazo[4,5,l-jk][l,4]benzodiazepine-2(lH)-thione monohydrochloride; mp. >280°C

25 (decomp.); [αj -, = -32.7° (cone. = 1.0% in methanol) (comp. 7)

Example 11

To a cooled (0°C) mixture of 0.75 parts of trichloromethyl chloroformate and 33.2 parts of dichloromediane under argon, there was added slowly a solution of 2.39 parts of intermediate 23 in 33.2 parts of dichloromethane. After stirring for 1/2 hour at 0°C, there was added NaHCO3 (sat). The organic layer was separated, dried, filtered and evaporated. The residue was purified by flash column chromatography (silica gel ;

CH2CI2 / CH3OH 98.5: 1.5). The eluent of die desired fraction was evaporated and the residue was crystallized from acetonitrile. The product was filtered off and dried, yielding 1.12 parts (44.1%) of (+)-(S)-8-chloro-6-(3-ethyl-2-pentenyl)-4,5,6,7- tetr^ydro-5-methyIimidazo[4,5,l-jk][l,4]benzodiazepin-2(lH)- one; mp. 156.8°C;

20 [o]J = +7.15° (c = 0.1% in methanol) (comp. 18).

Example 12

1.60 Parts of intermediate 27, 31.6 parts of ethanol and 3.37 parts of diiourea were combined under argon and refluxed overnight. The solvent was evaporated and the

residue was dissolved in l,l'-oxybisethane. This solution was washed with water, dried, filtered and evaporated. The residue was purified twice by flash column chromatography (silica gel ; CH2CI2 / CH3OH 99:1). The eluent of the desired fraction was evaporated and the residue was crystallized from acetonitrile. The product was filtered off and dried, yielding 0.55 parts (42.6%) of (+)-(S)-8-chloro-6-(3-ethyl-2- pentenyl)-4,5,6,7-tetrahydro-5-methylimidazo[4,5, 1 -jk][ 1 ,4] benzodiazepine-2( 1E)-

20 thione; mp. 113.5°C; [α] _D = +0.907° (c = 0.1% in methanol) (comp. 19).

Example 1 1.0 Parts of intermediate 24, 17.8 parts of tetrahydrofuran and 0.71 parts of 1,1'- cai >onothionylbis(lIi-in idazole) were combined under argon and refluxed for 1/2 hour. The solvent was evaporated and the residue was dissolved in dichloromediane. This solution was washed widi water, dried, filtered and evaporated. The residue was purified by flash column chromatography (silica gel ; CH2CI2 / CH3OH 95:5). The eluent of the desired fraction was evaporated and the residue was crystallized from methanol. The " product was filtered off and dried, yielding 0.50 parts (43.9%) of (+)-(S)-8-chloro-6-(2- cyclopentylideneethyl)-4,5,6,7-tetrahydro-5-methylimidazo[4, 5, 1 -jk] [ 1 ,4]-benzodiaze-

20 pine-2(lH)-thione; mp. 203.4°C; [α]^ = +15.44° (c = 1% in DMF) (comp. 22).

Example 14

To a mixture of 0.33 parts of trichloromethyl chloroformate and 20 parts of dichloromethane under argon there were added 1.0 part of intermediate 24 and dropwise a solution of 0.66 parts of jN-methylmorphoUne in 20 parts of dichloromethane. After stirring for 1/2 hour at 0°C, there was added NaHCO3 (sat.) and the whole was stirred for 1 hour at room temperature. The organic layer was separated, dried, filtered and evaporated. The residue was purified by flash column chromatography (silica gel ; CH2CI2 / CH3OH 98:2). The eluent of the desired fraction was evaporated and the residue was converted into the hydrochloride salt in 2-propanol by addition of l,l'-oxybisethane saturated with HCl. After cooling, the salt was filtered off and dried, yielding 0.31 parts (25.7%) of (-)-(S)-8-chloro-6-(2-cyclopentylideneethyl)-4,5,6,7- tetrahydiO-5-mediylimidazo[4,5,l-jk][l,4]benzodiazepin-2(lH) -one monohydro-

20 chloride; mp. 220.0°C; [αjj = -23.91° (c = 0.1% in methanol) (comp. 23).

All compounds listed in Table 1 were prepared following methods of preparation described in Examples 7-14, as is indicated in the column Ex. No.

Table 1

C. Pharmacological example Example 15

A rapid, sensitive and automated assay procedure was used for the in-vitro evaluation of anti-HTV agents. An HIV-1 transformed T4-cell line, MT-4, which was previously - shown (Koyanagi et al., Int. J. Cancer, 26, 445-451, 1985) to be highly susceptible to and permissive for HTV infection, served as die target cell line. Inhibition of the HIV- induced cytopatiiic effect was used as the end point. The viability of both HTV- and mock-infected cells was assessed spectrophotometrically via the in-situ reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The 50% cytotoxic dose (CD50 in μg ml) was defined as the concentration of compound tiiat reduced the absorbance of the mock-infected control sample by 50%. The percent protection achieved by the compound in HTV-infected cells was calculated by the following formula :

(ODT)HIV - (ODC)HIV expressed in %,

(OD _C )MOCK - (OD _C )HIV whereby (ODj _jj iy is the optical density measured widi a given concentration of die test compound in HTV-infected cells; (ODc)|flv is the optical density measured for the control untreated HTV-infected cells; (ODc) ocκ * ^{S me} optical density measured for the control untreated mock-infected cells; all optical density v.alues were determined at 540 nm. The dose achieving 50% protection according to the above formula was defined as die 50% effective dose (ED50 in μg/ml). The ratio of CD50 to ED50 was defined as the selectivity index (ST).

Table 2 : 50% cytotoxic (CD50), 50% effective dose (ED50) and selectivity index (SI).

P, Com osition Exam les

Example 16 : ORAL SOLUTION

9 g of methyl 4-hydroxybenzoate and 1 g of propyl 4-hydroxybenzoate are dissolved in 41 of boiling purified water. In 3 1 of this solution are dissolved first 10 g of 2,3-di- hydroxybutanedioic acid and diereafter 20 g of the A.I. The latter solution is combined widi the remaining part of the former solution and 121 1,2,3-propanetriol and 3 1 of sorbitol 70% solution .are added diereto.40 g of sodium s.accharin are dissolved in 0.51 of water and 2 ml of raspberry and 2 ml of gooseberry essence are added. The latter solution is combined widi die former, water is added q.s. to a volume of 201 providing .an oral solution comprising 5 mg of die active ingredient per teaspoonful (5 ml). The resulting solution is filled in suitable containers.

Example 17 : CAPSULES

20 g of the A.I., 6 g sodium lauryl sulfate, 56 g starch, 56 g lactose, 0.8 g colloidal silicon dioxide, and 1.2 g magnesium stearate are vigorously stirred together. The resulting mixture is subsequently filled into 1000 suitable hardened gelatin capsules, comprising each 20 mg of d e active ingredient.

Example 18 : FILM-COATED TABLETS

.pjejarap n oI.»M,e,t gore A mixture of 100 g of the A.I., 570 g lactose and 200 g starch is mixed well and diereafter humidified wid a solution of 5 g sodium dodecyl sulfate and 10 g poly- vinylpyrrolidone (Kollidon-K 90 ®) in about 200 ml of water. The wet powder mixture is sieved, dried -and sieved again. Then there are added 100 g microcrystalline cellulose (Avicel ®) -and 15 g hydrogenated vegetable oil (Sterotex ®). The whole is mixed well and compressed into tablets, giving 10.000 tablets, each containing 10 mg of the active ingredient.

Coating

To a solution of 10 g methyl cellulose (Mediocel 60 HG®) in 75 ml of denaturated ethanol tiiere is added a solution of 5 g of ethyl cellulose (Ethocel 22 cps ®) in 150 ml of dichloromethane. Then tiiere are added 75 ml of dichloromediane and 2.5 ml 1,2,3-propanetriol. 10 g of polyethylene glycol is molten and dissolved in 75 ml of dichloromethane. The latter solution is added to the former and dien there are added 2.5 g of magnesium octadecanoate, 5 g of polyvinylpyiTolidone and 30 ml of concentrated colour suspension (Opaspray K- 1-2109®) and die whole is homogenated. The tablet cores are coated with the tiius obtained mixture in a coating apparatus.

Example 19 : INJECTABLE SOLUTION

1.8 g methyl 4-hydroxybenzoate and 0.2 g propyl 4-hydroxybenzoate are dissolved in about 0.5 1 of boiling water for injection. After cooling to about 50°C there are added while stirring 4 g lactic acid, 0.05 g propylene glycol and 4 g of the A.I.. The solution is cooled to room temperature and supplemented with water for injection q.s. ad 1 1, giving a solution comprising 4 mg/ml of A.I.. The solution is sterilized by filtration (U.S. P. XVII p. 811) and filled in sterile containers.