HU208139B - Process for producing 5,11-dihydro-6h-dipyrido/3,2-b:2',3'-//1,4/ diazepines and pharmaceutical compositions comprising same - Google Patents

Abstract

The present invention relates to novel 5,11-dihydro-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepines. According to the invention, the compounds of formula (I) or their pharmaceutically acceptable acid addition salts are prepared wherein Z is oxygen or sulfur, R2 is C1-C5 alkyl or C3-7cycloalkyl, R3 is hydrogen or C1-C4 alkyl, and R5 is C1-C4 alkyl or trifluoromethyl. The invention further relates to a method for the manufacture of a medicament for the prevention or treatment of HIV-1 infections containing the above compounds as active ingredients. EN 208139 Scope of the description: 12 pages (including 2 sheets)

Description

The present invention relates to novel 5,11-dihydro-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepines, their pharmaceutically acceptable acid addition salts, and to the prevention of HIV infections containing these compounds. and pharmaceutical compositions for treatment thereof.

Human disease, Acquired Immunodeficiency Syndrome (AIDS), is caused by the HIV virus (Human Immunodeficiency Virus), primarily a strain known as HIV-1.

Like other viruses, HIV-1 virus cannot replicate without using the biosynthetic apparatus of the host cell it infects. This apparatus is used to produce the structural proteins that make up the virus. These proteins are encoded by the genetic material of the infectious viral particle, the virion. However, since it is a retrovirus, the genetic material for HIV is RNA, not DNA, as in the host cell genome. Therefore, viral RNA must first be transformed into DNA and then incorporated into the host cell genome in order for the host cell to produce the required viral proteins.

The conversion of RNA to DNA is accomplished by the use of the reverse transcriptase (RT) enzyme, which is present with the RNA in the infectious virion. Reverse transcriptase has three enzymatic functions: it acts as an RNA-dependent DNA polymerase, as a ribonuclease, and as a DNA-dependent DNA polymerase. First, as an RNA-dependent DNA polymerase, reverse transcriptase produces a single-stranded copy of viral RNA. Then acting as a ribonuclease, the reverse transcriptase frees the DNA being produced from the original viral RNA and then destroys the original RNA. Finally, acting as a DNA dependent DNA polymerase, the reverse transcriptase produces a second complementary DNA strand, using the first DNA strand as a template. The two strands form double-stranded DNA, which is integrated into the genome of the host cell by another enzyme called integrase.

Compounds that inhibit the enzyme functions of HIV-1 reverse transcriptase will inhibit HIV-1 replication in infected cells. These compounds are useful in preventing or treating HIV-1 infection in humans.

The present invention relates to novel 5,1-dihydro-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepines and their pharmaceutically acceptable acid addition salts of formula (I):

Z is oxygen or sulfur,

R ^{2 is C} 1-5 alkyl or C 3-7 cycloalkyl,

R ^{3 is} hydrogen or C 1-4 alkyl; and R ^{5 is C} 1-4 alkyl or trifluoromethyl.

It is advantageous

5,11-Dihydro-11-ethyl-4-methyl-6H-dipyrido [3,2-b: 2 ', 3'e] [1,4] -diazepin-6-one, 1-cyclopropyl-5,1 1-dihydro-4-methyl-6H-dipyrido [3,2b: 2 ', 3'-e] [1,4] diazepin-6-one or -thione;

5,11-Dihydro-11-ethyl-2,4-dimethyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-thione, 1-cyclopropyl-5 , 11-dihydro-2,4-dimethyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepin-6-one or -thione.

The compounds of formula (I) and their salts may be prepared by methods known per se or by obvious modifications thereof. The following procedures a) to c) illustrate the preparation of the compounds.

(a) Procedure

Compounds of formula (Ia) - R ² , R ³ and R ⁵ as defined above are prepared by reacting a carboxylic acid amide of formula (II) - wherein R ² , R ³ and formula (Ia) are as defined above. and Hal is cyclized with fluorine, chlorine, bromine or iodine.

The cyclization is conveniently carried out by converting the compound of formula II into its alkali metal salt and then condensing at a temperature between 0 ° C and the boiling point of the reaction mixture. At least moles of the metal compound must be used. Preferably, the metal salt is lithium, sodium or potassium hydride or an alkyl lithium such as n-butyllithium.

The cyclization reaction is usually carried out in an inert solvent such as tetrahydrofuran, 1,4-dioxane, glycol dimethyl ether, diethylene glycol dimethyl ether, triethylene glycol dimethyl ether, dimethylformamide, benzene or anisole. The cyclization can also be carried out by heating the carboxylic acid amide (II) in a dipolar aprotic solvent, preferably sulfolane or dimethylsulfone. The use of catalytic amounts of strong acids such as sulfuric acid, hydrochloric acid, hydrobromic acid, phosphoric acid, polyphosphoric acid, methanesulfonic acid or p-toluenesulfonic acid has proven to be useful. The required reaction temperature is usually 110 to 220 ° C.

(b) procedure

(I) a compound of the formula: - wherein Z is oxygen and R ^2, R ⁵ are as hereinbefore defined, are prepared by a general formula (Ib)

5,11-Dihydro-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepin6-one to the corresponding metal salt of Formula VIa or a compound of Formula VIb in the formulas M ^{+ is} an alkali metal such as lithium, sodium, potassium, rubidium or cesium ions or M ^{+ is} a group of formula MgHal ^{+ in} which Hal is chlorine, bromine or iodine, and the compound thus obtained is converted into ), wherein R ^{2 is} as defined above and X is a reactive group such as halogen, ester or sulfinyloxy.

Conversion of a compound of formula (Ib) to the corresponding alkali metal compound of formula (VIa) or (VIb) may be accomplished by reacting a compound of formula (Ib) with an alkyl lithium compound such as n-butyllithium or tert-butyllithium. in the case of tetramethylene diamine, a lithium dialkylamide such as lithium diisopropylamide, lithium dicyclohexylamide or lithium isopropylcyclohexylamide, an aryl lithium compound such as phenyl lithium, an alkali metal 2

20 hydroxide such as lithium, potassium or sodium hydroxide, an alkali metal hydride such as sodium or potassium hydride, an alkali metal amide such as sodium or potassium amide or a Grignard reagent such as methyl in the presence of magnesium iodide, ethyl magnesium bromide or phenyl magnesium bromide. One equivalent of base is required for the preparation of compound (VIa), while two equivalents are required for the preparation of compound (VIb). The metal compound is conveniently formed in an inert organic solvent at a temperature between -78 ° C and the boiling point of said reaction mixture. When an alkyl lithium, aryl lithium compound, lithium dialkyl amide or Grignard reagent is used to form the metal compound, the preferred solvents are ethers such as tetrahydrofuran, diethyl ether or dioxane, optionally with aliphatic or aromatic hydrocarbons, e.g. mixed with hexane or benzene, and the reaction is carried out at a temperature of -20 ° C to +80 ° C. When the metal compound is formed with an alkali metal hydride or an alkali metal amide, in addition to the solvents mentioned above, xylene, toluene, acetonitrile, dimethylformamide or dimethylsulfoxide may be used, whereas if an alkali metal hydroxide is used, alcohols such as ethanol or methanol ketones, such as acetone, and aqueous mixtures thereof.

The resulting alkali metal salt is converted to the compound of formula (I) by reacting the solution or suspension of the alkali metal compound with a compound of formula (VII) directly, i.e. without isolating the reaction product at -20 ° C to the reflux temperature of the reaction mixture.

The carboxylic acid amides of formula (II) used as starting materials for process (a) are prepared by, for example, a 2-chloronicotinic acid amide of formula (VIII) wherein R ³ , R ⁵ and Hal are as defined above primary amine: - wherein R ² is as defined above - is aminated. For example, compounds of formula Ib may be prepared in a manner analogous to process a).

The reaction may also be carried out in the presence of inorganic or organic auxiliary bases such as triethylamine, Ν, Ν-dimethylaniline or sodium or potassium carbonate.

The reaction may be carried out without the use of a solvent, but inert organic solvents are preferably employed at a temperature in the range of 0 to 175 ° C, preferably at reflux. Suitable inert solvents which may be used include excess primary amines of formula IX, open or cyclic ethers such as tetrahydrofuran, 1,4-dioxane, glycol dimethyl ether, diethylene glycol dimethyl ether; aromatic hydrocarbons such as benzene, toluene, xylene, chlorobenzene or pyridine; alcohols such as methanol, ethanol, isopropanol; dipolar aprotic solvents such as dimethylformamide; 1,3-dimethyl-2-imidazolidinone, 1,3-dimethyl-tetrahydro-2 (1H) -pyrimidinone and sulfolane.

Each of the several starting materials can be prepared by methods known in the literature, commercially available, or known in the literature.

c) procedure

According to process c), compounds of formula I wherein Z is sulfur are prepared by reacting a compound of formula I wherein Z is oxygen, such as 2,4-bis (4- methoxyphenyl) -1,3-dithia-2,4-diphosphetane-2,4-disulfide, bis (tricyclohexyltin) sulfide, bis (tri-n-butyltin) sulfide, bis (triphenyltin) sulfide, with bis (trimethylsilyl) sulfide or phosphorus pentasulfide. The reaction is carried out in an inert organic solvent such as carbon disulfide, benzene or toluene at room temperature or higher, preferably to the boiling point of the reaction mixture, and preferably under anhydrous conditions. When the aforementioned tin or silyl sulfides are used, the sulfurization reaction is preferably carried out in the presence of a Lewis acid such as boron trichloride.

If desired, the compounds of formula I may be converted into their non-toxic pharmaceutically acceptable acid addition salts by known methods, for example by dissolving a compound of formula I in a suitable solvent and acidifying the solution with one or more molar equivalents of the required acid. The present invention also relates to the preparation thereof.

Inorganic and organic acids which form non-toxic pharmaceutically acceptable acid addition salts with compounds of formula I include hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, methanesulfonic acid and the like. The compounds of formula I may form acid addition salts with 1 molar equivalent of acid.

The compounds of formula (I) described above have inhibitory activity against reverse transcriptase of HIV-1 virus. When administered in suitable dosage forms, the compounds are useful for the prophylaxis or treatment of AIDS, ARC, and the like, which are associated with HIV-1 infection.

The compounds of formula (I) may be administered in a single dose or in divided doses, orally, parenterally or topically. A suitable oral dose of the compounds of formula I is about 0.5 mg / g daily. For parenteral formulations, the appropriate dosage unit is 0.1 to 250 mg of the above compound, whereas for topical administration, the formulations preferably contain 0.01 to 1% of the active ingredient. It should be noted, however, that the dose administered will vary according to the patient, and the dose of a particular patient will depend on the judgment of the physician, who will determine the appropriate dose according to the size and condition of the patient and the patient's response to the drug.

When the compounds of the present invention are administered orally, they are administered as medicaments in the form of pharmaceutical compositions containing the active compounds in admixture with compatible pharmaceutical carriers. Such carriers are inert, organic or inorganic carriers suitable for oral administration. The carrier includes, for example, water, gelatin, talc, starch, magnesium stearate, acacia, vegetable oils, polyalkylene glycols, petroleum jelly and the like.

HU 208 139 Β

The pharmaceutical compositions may be prepared by conventional procedures. The finished dosage forms may be solid dosage forms such as tablets, dragees, capsules and the like; or liquid dosage forms such as solutions, suspensions, emulsions and the like. The pharmaceutical compositions may also be subjected to conventional pharmaceutical operations such as sterilization. The pharmaceutical compositions may further comprise conventional additives such as preserving, stabilizing, emulsifying, flavoring and wetting agents, buffers, salts, osmotic pressure modifying agents and the like. Suitable solid carriers include starch, lactose, mannitol, methylcellulose, microcrystalline cellulose, talc, silica, calcium phosphate (dibasic), and high molecular weight polymers such as polyethylene glycols.

For parenteral administration, the compounds of Formula I may be administered in aqueous or non-aqueous solutions, suspensions or emulsions, in pharmaceutically acceptable oils or liquid mixtures, which may contain bacteriostatic agents, antioxidants, preservatives, buffers or other solutes to form a solution in the blood. and thickening agents, suspending agents or other pharmaceutically acceptable additives. Examples of such additives are tartrate, citrate and acetate buffers, ethanol, propylene glycol, polyethylene glycol, complexing agents such as EDTA, antioxidants such as sodium bisulfite, sodium metabisulfite and ascorbic acid, high molecular weight pol liquid polyethylene oxides for viscosity control; and polyethylene derivatives of sorbitan anhydrides. If necessary, preservatives such as benzoic acid, methyl or propyl paraben, benzalkonium chloride or other quaternary ammonium compounds may be used.

The compounds of the present invention may also be used as solutions for nasal administration, and may contain, in addition to the compounds of the present invention, suitable buffers, tonicity regulators, antimicrobial preservatives, antioxidants, and viscosity enhancers in aqueous media.The viscosity enhancing agents include polyvinyl alcohols, derivatives, polyvinylpyrrolidones, polysorbates and glycerol. The antimicrobial preservative used may be benzalkonium chloride, thimerosal, chlorobutanol or phenylethyl alcohol.

The compounds of the present invention may also be administered in the form of suppositories.

As mentioned above, the compounds of the present invention inhibit HIV-1 reverse transcriptase enzyme activity. Based on the assay of the compounds described below, they are known to inhibit the RNA-dependent DNA polymerase activity of HIV-1 reverse transcriptase. It is known (data not reported) that the compounds also inhibit the DNA-dependent DNA polymerase activity of HIV-1 reverse transcriptase.

Using the reverse transcriptase assay described below, compounds can be tested for their ability to inhibit the RNA-dependent DNA polymerase activity of HIV-1 reverse transcriptase. Thus, certain specific compounds were tested according to the examples below. The results of the tests are shown in Table I below.

Reverse transcriptase (RT) assay

Theory of investigation

Among the enzymes encoded by the human immunodeficiency virus (HIV-1) is a reverse transcriptase (Benn, S. et al., Science, 230, 949 (1985)), which is called because it transcribes a copy of DNA. from an RNA temple. This activity can be quantitatively measured in a cell-free enzyme assay previously described (Farmeric, WG et al., Science 236, 305 (1987)), and is based on the observation that reverse transcriptase is capable of producing a synthetic template [poly (C) , initiated by oligo d (G)] to transcribe a radiolabeled acid-precipitated DNA strand using ³ H-dGTP as a substrate.

Materials

(a) Preparation of the enzyme

Reverse transcriptase from the human immunodeficiency virus (HIV-1) LAV strain (LAV = lymphadenopathy virus) (Benn, S. et al., Science, 230, 949 (1985)) was isolated from the bacterial strain JM109 [Yanisch-Perron, C. Viera, J. and Messing, J., Gene, 33, 103 (1985)], expressing the DNA clone pBRTprtl + (Farmeric, WG et al., Science, 236, 305 (1987)), which is under the control of the lac promoter. pIBI21 expression vector [International Biotechnologies, Inc. New Haven, CT 06535]. The culture was supplemented overnight with 100 pg / ml ampicillin in 2XYT medium (Maniatis,

T., Fritsch, EF and J. Sambrook eds .: Molecular Cloning: A laboratory manuel; Cold Spring Harbor Laboratory (1982), and inoculated at a 1:40 dilution in M9 medium (Maniatis, T., Fritsch, EF, and 10 µg / ml thiamine, 0.5% casein amino acids, and 50 pg / ml ampicillin) for positive selection. J. Sambrook, Molecular Cloning: A Laboratory Manual; Cold Spring Harbor Laboratory, 1982] The culture was incubated at 37 ° C (225 rpm) until its optical density, OD _50, reached 0.3. 0.4 The IPTG (isopropyl-PD-thiogalactopyranoside) repressor inhibitor was added to 0.5 mM and incubated for an additional 2 hours. The bacteria were pelleted with 50 mM Tris buffer, 0.6 mM EDTA and 0.375 M NaCl. and lysozyme (1 mg / ml) was digested on ice for 30 minutes, cells were lysed with 0.2% NP-40 (nonidet P-40) and adjusted to 1M NaCl.

Insoluble cellular debris is removed by centrifugation and the protein precipitated by addition of 3 volumes of saturated aqueous ammonium sulfate. The enzyme was pelleted, resuspended in RT buffer (50 mM Tris, pH 7.5; 1 mM EDTA; 5 mM DTT, 0.1% NP-40; 0.1 M NaCl, and 50% glycerol) and -70 Store at ° C.

b) Composition of 2x concentrated strain reaction mixture

HU 208 139 Β

Strain Reagent 2x concentrated mixture

1M Tris pH 7.4 100 mM

1M dithiothreitol 40 mM

1M NaCl 120 NmM

1% Nonidet P-40 0.1%

1M MgCl 4 mM [poly (C) / oligo d (G)] 5: 1 2 pg / ml ³ H-dGTP (81 μΜ) 0.6 μΜ

Execution of the test

The 2x concentrated strain reaction mixture was aliquoted and stored at -20 ° C. The mixture is thus stable and thawed for use in each assay. This enzyme assay was applied to a 96-well microtiter plate system and has already been described in the literature (Spira, T. et al., J. Clinical Microbiology, 25, 97 (1987)). Tris buffer (50 mM, pH 7.4), solvent (diluted solvent to suit dilution of the compound), or compound in solvent was distributed into 96-well microtiter plates (10 μΐ / well; 3 wells per compound).

The HIV-1 reverse transcriptase enzyme is thawed, diluted in 50 mM Tris buffer, pH 7.4, so that 15 μΐ of the diluted enzyme contains 0.001 units (1 unit of enzyme per minute at 25 ° C). on 1 μΜ substrate) and place 15 μΐ in a container. Add 20 μΐ 0.12-0.5 M EDTA to the first three wells of the microtiter plate. EDTA chelates the Mg 2 present and prevents reverse transcription. This group serves as a basic polymerization and is subtracted from all other groups. To each well was added 25 μΐ 2x concentrated reaction mixture and incubated at room temperature for 60 minutes. At the end of the assay, the DNA is precipitated in each well with 50 μΐ of 10% trichloroacetic acid in 1% sodium pyrophosphate. The microtiter plate is incubated at 4 ° C for 15 minutes and the precipitate is applied to # 30 glass fiber paper (Schleicher-Schuell) using a "Skatron semi-automatic harvester". The filters are then washed with additional 5% trichloroacetic acid containing 1% sodium pyrophosphate, rinsed with 70% aqueous ethyl alcohol, dried and transferred to scintillation vials (Spira T. et al., J. Clinical Microbiology, 25, 97). 1987)]. Add 2 ml of scintillation cocktail to each vial and count on a Beckman β-counter. The percentage inhibitory effect is calculated as follows:

inhibitory%

CPM Mean Test Value - CPM Mean Control Valuex100

CPM mean control value

In order to demonstrate that the compounds which are active in the reverse transcriptase assay also have the ability to inhibit HIV replication in a living system, the compounds of the present invention were also tested in the human T cell culture assay described below. . The results of this assay are illustrated in Table I.

Human T cell culture assay

Theory of investigation

Syncytia formation is a characteristic of in vitro cultures of CD4 + T cells infected with HIV-1 virus. In this assay, T cells are treated with a compound that is thought to be an inhibitor of replication and then infected with HIV-1 virus. After incubation, the culture is examined for syncytia formation. Lack of syncope or decrease in number of syncytes is used as a measure of the ability of the test compound to inhibit HIV replication.

Test procedure

Target cells were AC 8166, which have the introduced human lymphoma T-cell-derived cells, a subclone of 5 x ⁴ / LOO μΐ initial RPMI containing 10% fetal bovine serum in 1640 medium in 96 well (flat bottom) plates and a solution of the test compound dissolved in dimethyl sulfoxide, chosen amount. After 24 hours, each culture was inoculated with HIV-1 strain HTLV-IIIB (GM Shaw, RH Hahn, SK Arya, JE Groopman, RC Gallo and F. Wong-Staall, Science 226: 1165 (1984)) 50-100 TCID. ₅₀ doses (the dose that produces effect in 50% of the cultures; TCID = tissue culture infective dose). Control cultures receive only compound or virus only. Four days after the viral infection, the cultures are visually examined for viral giant syncytia frequency and distribution. Percent inhibitory activity of the test compound is determined relative to control values. The presence or absence of viral replication was confirmed by harvesting cell-free culture fluids from each experimental group and, after 3 days, determining the presence or absence of infectious growth by syncytial formation in secondary human T-cell cultures.

In order to determine the specificity of the enzyme inhibitory activity of the compounds of the invention, some are tested for inhibitory activity against reverse transcriptase from feline leukemia virus and calf-derived DNA polymerase using methods known per se. None of the compounds tested in this way showed inhibitory activity against these enzymes. These results indicate that the compounds of the present invention are sufficiently specific for their enzyme inhibitory activity against HIV reverse transcriptase.

To roughly determine the cytotoxicity of the compounds of the present invention, some of these compounds were assayed in the MTT cellular cytotoxicity assay described below. The result of the investigation5

EN 208 139 Β are shown in Table I below. Compounds having relatively high ED ₅₀ values are preferred.

MT MTT assay for cellular cytotoxicity

Theory of investigation

The MTT [3- (4,5-Dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide] assay is based on the cleavage of tetrazolium bromide by metabolically active cells, resulting in a high quantitative blue color. This assay has been previously described [Mosmann, Tim, J. Immunol. Methods, 65, 55 (1983)] but optimized for the following test.

Test method

The target cell line in the assay is the H9 cell line [Jacobs, JP: J. Natl. Cancer Inst., 34, 231 (1965)], a suspension of a standard human lymphoma cell line grown in RPMI 1640 medium supplemented with 10% fetal bovine serum. Cells (100 µl) were placed in wells of a microtiter plate at a concentration of 10 ⁵ cells / ml in the presence of various concentrations of inhibitor. Cells were incubated in a humidified carbon dioxide incubator at 37 ° C. After 5 days, 20 µl of MTT (5 mg / ml in RPMI 1640, sonicated, filtered through a 0.2 µ filter and stored at 4 ° C) was added to each well. After an additional 4 hours incubation at 37 ° C, 60 µl of triton-X was added to each well and mixed thoroughly to facilitate solubilization of the crystals. 5 µl of anhydrous ethanol was then added to each well, and the resulting mixture was incubated for 30 minutes at 60 ° C, and the color change was read immediately on a Dynatech plate reader at 570 nm. Data from the assay were used for non-linear regression analysis to obtain ED50 values. spreadsheet

Compound (example number) Port. Tr Test% Inhibition 10 Rg / ml T-cell assay% inhibition 3 pg / ml Cytotoxicity assay (ED ₅₀ , μΜ) First 100 NV NV Second 100 100 200 Third 100 100 250 4th 75 ^x NV NV 5th 85 ^x NV NV 6th 76 ^x NV NV

Remarks: ^x 1 μΜ NV was not tested

The invention is illustrated by the following examples, without limiting the scope of the invention to the specific examples.

Example 1

5,11-Dihydro-11-ethyl-2-methyl-4-trifluoromethyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepin-6-one

a) 3-Cyano-2-hydroxy-6-methyl-4- (trifluoromethyl) pyridine

A solution of cyanoacetamide (14.0 g) in ethanol (80 ml) was heated to 50 ° C, and piperidine (14 g) and trifluoroacetylacetone (25 g) were added. The resulting mixture was stirred at 70 ° C for 30 minutes and then at room temperature overnight. The reaction mixture was concentrated in vacuo and the residue diluted with water (100 mL). 15 ml of concentrated hydrochloric acid are carefully added to the reaction mixture while stirring. After 15 minutes, the precipitate was filtered off and dried in vacuo overnight to give 27.8 g of the title cyanopyridine.

b) A mixture of 3-aminocarbonyl-2-chloro-6-methyl-4- (trifluoromethyl) pyridine in ml of phosphorus trichloride and 9.8 g of the above cyanopyridine was heated at reflux for 5 hours. The mixture was then cooled and cautiously quenched with 400 mL of ice water. The product was extracted with dichloromethane, washed with saturated sodium bicarbonate solution and dried over anhydrous magnesium sulfate. The solution is filtered and concentrated in vacuo, the crude chlorine compound is dissolved in 50 ml of concentrated sulfuric acid and heated to 140 ° C for 20 minutes. The mixture was then cooled, carefully poured onto 600 mL of ice, the precipitate was filtered off, washed with ice water and dried to give 7.6 g of the desired amide. The filtrate was extracted with ethyl acetate (200 mL), dried over anhydrous magnesium sulfate, filtered and concentrated to give an additional 1.7 g of product.

c) 3-Amino-3-chloro-6-methyl-4- (trifluoromethyl) pyridine

6.6 g of sodium hydroxide are dissolved in 60 ml of water and 9.3 g of bromine are added at 5 ° C. When the solution is clear, 9.2 g of 3-aminocarbonyl-2-chloro-6-methyl-4- (trifluoromethyl) pyridine are added rapidly while maintaining the temperature below 5 ° C. The resulting mixture is stirred until the 3-aminocarbonylpyridine is dissolved (about 30 minutes). At this time, the cooling bath was removed and the reaction mixture was heated to 75 ° C for 30 minutes.

The mixture was cooled to room temperature, the amino-pyridine product 3 was extracted with ethyl acetate, dried over anhydrous magnesium sulfate, filtered and evaporated to give 4.9 g of the desired product.

d) 2-Chloro-N- (2-chloro-6-methyl-4-trifluoromethyl-3-pyridinyl) 3-pyridine carboxamide

2.1 g of 3-amino-2-chloro-6-methyl-4-trifluoromethyl) pyridine are dissolved in 10 ml of tetrahydrofuran and 7 ml of 1.5 M cyclohexane lithium are added to the solution at -78 ° C over 3 minutes. diisopropylamine solution is added dropwise. The reaction mixture is stirred for 5 minutes and 0.9 g of 2-chloronicotinic acid chloride in 3 ml of tetrahydrofuran is added over 1 minute. After 5 minutes, a further 3 ml of lithium diisopropylamine solution was added, followed by another 0.5 g of acid chloride in 1 ml of tetrahydrofuran. The resulting mixture was stirred for 10 minutes and quenched with water (100 mL). Ethyl acetate (30 mL) was added to the organic layer, extracted with water, the combined aqueous layers were extracted with dichloromethane, dried over anhydrous magnesium sulfate, filtered and evaporated to give the crude product. It was washed with a little ethyl acetate and dried to give 1.3 g of the title compound.

HU 208 139 Β

e) N- (2-Chloro-6-methyl-4-trifluoromethyl-3-pyridinyl) -2-ethylamino-3-pyridine carboxamide

To a suspension of 2-chloro-N- (2-chloro-6-methyl-4-trifluoromethyl-3-pyridinyl) -3-pyridinecarboxamide (1.3 g) in xylene (5 ml) was added ethylamine (0.4 g), and the resulting mixture was heated in a pressure tube for 30 minutes at 160 ° C. The reaction mixture was cooled, diluted with ethyl acetate, washed, dried, and concentrated. The product was chromatographed on a silica gel column with ethyl acetate / hexane (1: 1) as eluent. 0.5 g of product is obtained.

f) 5,11-Dihydro-11-ethyl-2-methyl-4-trifluoromethyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-4 you

For 0.2 g of a 50% solution of sodium hydride in oil, 0.5 g of N- (2-chloro-6-methyl-4-trifluoromethyl-3-pyridinyl) -2-ethylamino dissolved in 3 ml of pyridine 3-Pyridine carboxamide was added. The reaction mixture was heated to 150 ° C, then cooled and concentrated in vacuo. Water was added to the residue, the product was extracted with ethyl acetate, dried over anhydrous magnesium sulfate, filtered and evaporated. The product was purified by silica gel column chromatography eluting with dichloromethane followed by dichloromethane / methanol. The solution was concentrated in vacuo and the residue crystallized from hexane to give 0.09 g of the title compound, m.p. 150-151 ° C.

Example 2

5,11-Dihydro-11-ethyl-4-methyl-6H-dipyrido [3,2-b:

2 ', 3'-e] [1,4] diazepin-6-one

a) A mixture of 2-chloro-4-methyl-3-nitropyridine g, 2-hydroxy-4-methyl-3-nitropyridine, 12.5 g phosphorus pentachloride and 62 ml phosphorus trichloride oxide was refluxed for 2 hours. The mixture was then cooled, poured into crushed ice and stirred until a precipitate formed. The product was extracted with dichloromethane, dried over anhydrous sodium sulfate and concentrated to give a brown oil which was washed with hot hexane and evaporated in vacuo. 16.2 g of the title compound are obtained, m.p. 45-47 ° C.

b) 3-Amino-2-chloro-4-methylpyridine

To 470 ml of acetic acid was added 16.2 g of 2-chloro-4-methyl-3-nitropyridine and the resulting mixture was stirred at room temperature for 15 minutes. Then, a solution of 160 g of tin chloride dihydrate in 200 ml of concentrated hydrochloric acid was added in one portion, and the resulting mixture was stirred overnight at room temperature. The mixture was diluted to 1 liter with water and 10N sodium hydroxide was slowly added with cooling until the white precipitate of tin hydrochloride dissolved. The product was extracted with dichloromethane, dried over anhydrous sodium sulfate and evaporated. 12.8 g of a yellow oil are obtained, which solidifies on standing. The product can be used in the following reaction, almost pure 3-amino-2-chloro-4-methylpyridine.

c) 2-Chloro-N- (2-chloro-4-methyl-3-pyridinyl) -3-pyridine carboxamide

The above carboxamide was prepared from 12.8 g of 3-amino-2-chloro-4-methylpyridine, 15.8 g of 2-chloronicotinic acid chloride, 7.1 g of pyridine, 30 ml of cyclohexane and 60 ml of dioxane. After removal of the solvent, the product was dissolved in dichloromethane, washed with water and dried over anhydrous sodium sulfate. The solvent was removed and the residue was washed with ethyl acetate to give 1.2 g of the title compound, m.p. 193-194 ° C.

d) N- (2-Chloro-4-methyl-3-pyridinyl) -2-ethylamino-3-pyridinecarboxamide

To a suspension of 21.0 g of 2-chloro-N- (2-chloro-4-methyl-3-pyridinyl) -3-pyridinecarboxamide in 150 ml of xylene is added 12.7 g of ethylamine in a steel bomb. The mixture was heated in an oil bath at 165 ° C for 6 hours and then stirred overnight at room temperature. The solvent was removed in vacuo and water was added to the residue. The product was extracted with ether and the extract was dried over anhydrous sodium sulfate and concentrated to give an oil. The oil was dissolved in ethyl acetate and hexane was added to give a precipitate. The solid was filtered off and dried, yielding 16.5 g of the title compound, m.p. 122-124 ° C.

e) 5,11-Dihydro-11-ethyl-4-methyl-6H-dipyrido [3,2-b:

2 ', 3'-e] [l, 4] diazepin-6-one

To a solution of 16.0 g of N- (2-chloro-4-methyl-3-pyridinyl) -2-ethylamino-3-pyridinecarboxamide (16.0 g) in 200 ml of dimethylformamide was added a 50% suspension of 7.9 g of sodium hydride. and stir the reaction mixture for 30 minutes. The mixture was heated at reflux for 2 hours, then cooled and carefully treated with crushed ice. The solvent was removed in vacuo and water was added to the residue. The product was extracted with ether, dried over anhydrous sodium sulfate and concentrated. The residue was boiled with ethyl acetate / cyclohexane (1: 1) and filtered to give 4.1 g of an almost pure product. 2.0 g of this product are further purified by recrystallization from dichloroethane to give 1.0 g of pure 5,11dihydro-11-ethyl-4-methyl-6H-dipyrido [3,2-b: 2 ' 3'-e] [1,4] diazepin-6-one, m.p. 212-214'C.

Example 3

-Cyclopropyl-5,11-dihydro-4-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepin-6-one

Following the procedure of Example 2, but using cyclopropylamine instead of ethylamine, the title compound is obtained, m.p. 247249C.

Example 4

5,11-Dihydro-11-ethyl-4-methyl-6H-dlpyrido [3,2-b:

', 3'-e] [1,4] diazepin-6-one

5.75 g (0.025 mol) of 5,11-dihydro-4-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepin-6-one were prepared with 100 ml of dimethylformamide. To this solution was added 2.00 g of a 50% sodium hydride dispersion in mineral oil. After hydrogen evolution ceased, the mixture was heated to 50 ° C for 30 minutes and then cooled to room temperature. 7.80 g of ethyl iodide (neat) are then added dropwise (15 minutes) to the reaction mixture and stirred overnight at room temperature.

The excess sodium hydride is decomposed by the careful addition of ice and water is added to the mixture.

HU 208 139 Β

The product was extracted with ether, dried over anhydrous sodium sulfate and evaporated. 4.5 g (70%) of 5,11-dihydro-11-ethyl-4-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepin-6-one are obtained. mp 212-214 ° C.

Example 5

In a manner similar to that described in the Examples above, the compound of Example II was prepared. Table 4-6. compounds.

II. spreadsheet

In the compounds of formula (I) in the Examples, R ² , R ³ and R ^{5 are as} defined below and Z is oxygen, unless indicated as sulfur.

Compound number R ² R ³ / R ⁵ Op. ('C) 4th cyclobutyl 4-Methyl-R ⁵ 214-215 5th cyclopropyl 2,4-Dimethyl-R ³ / R ⁵ > 300 6th cyclopropyl 4-ethyl-R ⁵ 228-230

Example 6

5,11-Dihydro-6H-11-cyclopropyl-4-methyldipyrido- [3,2-b: 2 ', 3' -e] [1,4] diazepine-6-thione

5.0 g (18.77 mmol) of 5,11-dihydro-6H-11-cyclopropyl-4-methyl-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6 and a mixture of 3.8 g (9.40 mmol) of dimeric p-methoxyphenylthienophosphine sulfide (Lawesson's reagent) in 100 mL of toluene is refluxed for 2.5 hours. The solution was then cooled to room temperature and allowed to stand overnight. The toluene was distilled off and the residue was purified by flash chromatography on silica gel with dichloromethane / ethyl acetate (6: 1). This gave 35 yellow oils which solidified on standing. The product was recrystallized from a mixture of ethyl ether and petroleum ether to give a bright yellow solid, which was dried at 80 ° C under high vacuum for 12 hours. 1.7 g (32%) of product are obtained, m.p. 189-194 ° C. 40

Basic composition:

Found: C, 63.81; H, 5.00; N, 19.84; S, 11.35; found: 63.75, 5.10, 19.88, 11.24%.

Examples for the preparation of pharmaceutical compositions

Example A)

Tablets or capsules

A-I THE 2 components How much- ness components How much- ness Example 3 250 mg Example 3 50 mg starch 160 mg dicalcium phosphate wood 160 mg mikrokr. cellulose 90 mg mikrokr. cellulose 90 mg Sodium starch glycolate 10 mg Sodium starch glycolate 10 mg

A-I THE 2 components How much- ness components How much- ness Magnesium steroid reap 2 mg stearic 5 mg Colloidal silica 1 mg Colloidal silica 1 mg

Production Process: The active ingredient and premixed ingredients - except for the lubricant - are prepared as a powder mixture. The lubricant is then blended and the resulting mixture is compressed into tablets or filled into hard gelatin capsules.

Example B Parenteral solution

components

The compound of Example 3 is tartaric acid benzyl alcohol water for injection

The amount is 500 mg

R5 g

0.1% by weight to 100 ml

Preparation method: The additives are mixed with water and the active ingredient is added. Stir the solution until it is clear. The solution was adjusted to pH 3.0, filtered into suitable vials or ampoules and autoclaved.

Example C Nasal drops

components

The compound of Example 3 is citric acid benzalkonium chloride

EDTA polyvinyl alcohol water

The amount is 100 mg

1.92 g

0.025% by weight 0.1% by weight

To 100 ml

Preparation procedure: Add the ingredients to the water, add the active ingredient and mix until the solution is clear. The pH of the solution is adjusted to 4.0 and filled into suitable vials or ampoules.

Claims (4)

A process for the preparation of 5,11-dihydro-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepines of the formula I and their pharmaceutically acceptable acid addition salts comprising the following: Z is oxygen or sulfur, R ^{2 is C} 1-5 alkyl or C 3-7 cycloalkyl, R ^{3 is} hydrogen or C 1 -C 4 alkyl, and R ^{5 is C} 1-4 alkyl or trifluoromethyl, characterized in that a) (I) Compounds of formula wherein Z is oxygen, R ^2, R ³ and R ⁵ are as defined above, Cyclizing a carboxylic acid moiety of formula II wherein R ² , R ³ and R ⁵ are as defined and Hal is fluorine, chlorine, bromine or iodine; obsession b) (I) Compounds of formula wherein Z is oxygen, R ^2, R ³ and R ⁵ are as defined above, (Ib), a compound of formula - R ³ and R ⁵ are as defined above - one (VIa) wherein or a metal salt of formula VIb - wherein M ^{+ is} an alkali metal such as lithium, sodium, potassium, rubidium or cesium or M ⁺ is a MgHal ⁺ group in which Hal is a chlorine, bromine or iodine ion - forming and alkylating the compound so obtained with a compound of formula VII wherein R ^{2 is} as defined above and X is a reactive group; obsession c) for the preparation of a compound of formula I wherein Z is sulfur, reacting a compound of formula I wherein Z is oxygen with a sulfurizing agent; and converting the resulting compound of formula I into a pharmaceutically acceptable acid addition salt thereof, if desired. The method of claim 1 5.11-Dihydro-11-ethyl-4-methyl-6H-dipyrido [3,2-b: 2 ', 3'e] [1,4] diazepin-6-one; 11-cyclopropyl-5,1-dihydro-4-methyl-6H-dipyrido [3,2b: 2 ', 3'-e] [1,4] diazepin-6-one or -thione; 11-Cyclopropyl-5,1-dihydro-2,4-dimethyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepin-6-one and its pharmaceutically acceptable salts. , characterized in that the appropriate starting compounds are reacted. The process according to claim 1, which is 11-cyclopropyl-5,11-dihydro-4-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepin-6-one or - or a pharmaceutically acceptable acid addition salt thereof, which comprises reacting appropriate starting materials. 4. A method comprising (I) compounds of the formula - Z, R ^2, R ³ and R ⁵ are defined as in claim 1 - or the pharmaceutical compositions containing a pharmaceutically acceptable acid addition salts thereof, wherein the active ingredient prepared according to claim 1, the pharmaceutical preparations may contain conventional together with carriers, diluents, fillers and / or other excipients, to form a pharmaceutical composition.