DE10312969A1 - Benzofuro-1,4-diazepin-2-one derivatives - Google Patents

Abstract

The present invention relates to new benzofuro-1,4-diazepin-2-one derivatives, processes for their preparation and their use as P2X¶4¶ receptor antagonists for the production of medicaments for the treatment and / or prophylaxis of diseases, in particular arteriosclerosis, Restenosis and other inflammatory diseases.

Description

The present invention relates to new benzofuro-1,4-diazepin-2-one derivatives, processes for their preparation and their use as P2X ₄ receptor antagonists for the production of medicaments for the treatment and / or prophylaxis of diseases, in particular arteriosclerosis, restenosis and other inflammatory diseases.

Atherosclerosis is a multifactorial disease that affects many different factors. Inflammatory processes play a central role here, in which inflammation-inducing cytokines such as CD40L and IFNγ are involved [P. Libby, Nature 420 (6917): 868-74 (2002)]. The purine receptor P2X ₄ belongs to the P2X family. So far, six different P2X receptors have been described in humans. These are calcium permeable channels that can be activated by ATP [F. Di Virgilio et al., Blood 97 (3): 587-600 (2001); RA North, A. Surprenant, Annu. Rev. Pharmacol. Toxico1. 40: 563-80 (2000)]. It could be shown that the P2X ₄ channel is highly expressed in highly vascularized organs and vessels [K. Yamamoto et al., Circ. Res. 87 (5): 385-91 (2000)]. Surprisingly, the P2X ₄ receptor is also expressed on human monocytes. When human monocytes were incubated with CD40L and IFNγ, a five-fold increase in P2X ₄ expression was observed. Also in the aortic vein of rabbits after damage by balloon angioplasty and cholesterol feeding [TJ Pulvirenti et al., J. Neurocytol. 29 (9): 623-31 (2000)] and in the arteriosclerotically altered vascular sections of the apoE knockout mouse a high expression of the P2X ₄ receptor was found. Since activated monocytes play a key role in the early stage of atherogenesis and restenosis and the activation of the monocytes by the cytokines mentioned, inhibition of activation leads to a reduction in atherogenesis [P. Libby, Nature 420 (6917): 868-74 (2002)]. Since the activation of monocytes by CD40L and IFNγ is obviously associated with an increase in P2X ₄ receptor expression and the associated increased calcium influx, blocking the P2X ₄ receptors should reduce the inflammatory processes [F. Di Virgilio, A. Solini, Br. J. Pharmacol. 135 (4): 831-42 (2002)]. Thus diseases in which inflammatory processes play a role could be treated via the blockade of the P2X ₄ receptors.

In addition to the indications atherosclerosis and restenosis and its complications (stroke, angina pectoris, heart attack, kidney failure, circulatory disorders of the Limbs) could hence other inflammatory Diseases such as Psoriasis and rheumatoid arthritis mentioned mechanism of treatment become accessible.

The synthesis of some benzofuro [3,2-e] -1,4-diazepin-2-one derivatives is described in J. Heterocyclic Chem. 16, 189-90 (1979) and ibid. 20, 1251-1254 (1983). Benzofuro-l, 4-diazepine derivatives with anti-ulcer effects are used in the EP 350 131-A disclosed.

in welcher
R¹ Halogen
und
R² Wasserstoff, Halogen, Nitro, Cyano oder eine Gruppe der Formel -C(O)-OR³, -C(O)-NR⁴R⁵, -SO₂-OR³ oder -SO₂-NR⁴R⁵, worin
R³, R⁴ und R⁵ unabhängig voneinander für Wasserstoff oder (C₁-C₆)-Alkyl stehen,
oder
R¹ Wasserstoff und
R² Halogen, Nitro, Cyano oder eine Gruppe der Formel -C(O)-OR³, -C(O)-NR⁴R⁵, -SO₂-OR³ oder -SO₂-NR⁴R⁵, worin
R³, R⁴ und R⁵ unabhängig voneinander für Wasserstoff oder (C₁-C₆)-Alkyl stehen,
bedeutet.The present invention relates to compounds of the general formula (I)

in which
R ¹ halogen
and
R ^{2 is} hydrogen, halogen, nitro, cyano or a group of the formula -C (O) -OR ³ , -C (O) -NR ⁴ R ⁵ , -SO ₂ -OR ³ or -SO ₂ -NR ⁴ R ⁵ , wherein
R ³ , R ⁴ and R ⁵ independently of one another represent hydrogen or (C ₁ -C ₆ ) alkyl,
or
R ^{1 is} hydrogen and
R ^{2 is} halogen, nitro, cyano or a group of the formula -C (O) -OR ³ , -C (O) -NR ⁴ R ⁵ , -SO ₂ -OR ³ or -SO ₂ -NR ⁴ R ⁵ , wherein
R ³ , R ⁴ and R ⁵ independently of one another represent hydrogen or (C ₁ -C ₆ ) alkyl,
means.

The compounds of the invention can also in the form of their salts, solvates and solvates of their salts.

In the context of the present invention, the substituents generally have the following meaning:
In the context of the invention, (C ₁ -C ₆ ) alkyl and (C ₁ -C ₄ ) alkyl stand for a straight-chain or branched alkyl radical having 1 to 6 or 1 to 4 carbon atoms. A straight-chain or branched alkyl radical having 1 to 4 carbon atoms is preferred.

Be exemplary and preferred called: methyl, ethyl, n-propyl, isopropyl and tert-butyl.

Halogen closes within the scope of the invention Fluorine, chlorine, bromine and iodine. Chlorine or bromine are preferred.

The compounds of the invention can in dependence of the substitution pattern in stereoisomeric forms, which are either like image and mirror image (enantiomers), or that don't like each other Behavior of image and mirror image (diastereomers) exist. The The invention relates to both the enantiomers or diastereomers also their respective mixtures. The racemic shapes can also be used as the diastereomers in a known manner in the stereoisomerically uniform Separate components.

Furthermore, certain connections exist in tautomeric forms. This is known to the person skilled in the art, and such compounds are also within the scope of the invention.

The compounds of the invention can also are present as salts. In the context of the invention are physiological safe salts preferred.

Physiologically acceptable salts can Salts of the compounds according to the invention with inorganic or organic acids. To be favoured Salts with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid or Sulfuric acid, or salts with organic carboxylic or sulfonic acids such as, for example, acetic acid, propionic acid, maleic acid, fumaric acid, malic acid, citric acid, tartaric acid, lactic acid, benzoic acid, or methane, Ethanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid or Naphthalenedisulfonic.

Physiologically acceptable salts can likewise salts of the compounds according to the invention with bases, such as metal or ammonium salts. preferred Examples are alkali metal salts (e.g. sodium or potassium salts), Alkaline earth salts (e.g. magnesium or calcium salts) as well as ammonium salts, which are derived from ammonia or organic amines, such as Ethylamine, di- or triethylamine, ethyldiisopropylamine, monoethanolamine, Di- or triethanolamine, dicyclohexylamine, dimethylaminoethanol, Dibenzylamine, N-methylmorpholine, dihydroabietylamine, 1-ephenamine, N-methylpiperidine, arginine, lysine, ethylenediamine or 2-phenylethylamine.

The compounds according to the invention and their salts can also in the form of their solvates, in particular in the form of their hydrates.

Compounds of the general formula (I) in which
R ¹ chlorine or bromine
and
R ^{2 is} hydrogen, chlorine, bromine, nitro, cyano or a group of the formula -C (O) -OR ³ or -C (O) -NR ⁴ R ⁵ , wherein
R ³ , R ⁴ and R ⁵ independently of one another represent hydrogen or (C ₁ -C ₄ ) -alkyl,
or
R ^{1 is} hydrogen
and
R ^{2 is} chlorine, bromine, nitro, cyano or a group of the formula -C (O) -OR ³ or -C (O) -NR ⁴ R ⁵ , wherein
R ³ , R ⁴ and R ⁵ independently of one another represent hydrogen or (C ₁ -C ₆ ) alkyl,
means.

in welcher
R¹ Chlor oder Brom
und
R² Wasserstoff, Chlor, Brom, Nitro oder Cyano,
oder
R¹ Wasserstoff
und
R² Chlor, Brom, Nitro oder Cyano bedeutet.Compounds of the general formula (Ia) are particularly preferred

in which
R ¹ chlorine or bromine
and
R ^{2 is} hydrogen, chlorine, bromine, nitro or cyano,
or
R ^{1 is} hydrogen
and
R ^{2 means} chlorine, bromine, nitro or cyano.

in welcher R¹ die oben angegebenen Bedeutungen hat,
in einem inerten Lösungsmittel in Gegenwart einer Base mit einer Verbindung der Formel (III)

in welcher R² die oben angegebenen Bedeutungen hat und
X¹ für eine geeignete Abgangsgruppe wie beispielsweise Chlor, Brom oder Iod steht,
zunächst zu Verbindungen der Formel (IV)

in welcher R¹ und R² die oben angegebenen Bedeutungen aufweisen,
umsetzt, diese dann unter zwischenzeitlicher Isolierung oder in einer Eintopfreaktion in Gegenwart einer Base zu Verbindungen der Formel (V)

in welcher R¹ und R² die oben angegebenen Bedeutungen aufweisen,
cyclisiert, anschließend in einem inerten Lösungsmittel in Gegenwart einer Base mit einer Verbindung der Formel (VI)

in welcher
X² und X³ gleich oder verschieden sind und für eine geeignete Abgangsgruppe wie beispielsweise Chlor, Brom oder Iod stehen, in Verbindungen der Formel (VII)

in welcher R¹, R² und X³ die oben angegebenen Bedeutungen aufweisen,
überführt, abschließend in einem inerten Lösungsmittel mit Ammoniak unter Cyclisierung umsetzt und die resultierenden Verbindungen der Formel (I) gegebenenfalls mit den entsprechenden Lösungsmitteln und/oder Basen oder Säuren in ihre Solvate, Salze und/oder Solvate der Salze überführt.In addition, a process for the preparation of the compounds according to the invention was found, characterized in that compounds of the formula (II)

in which R ^{1 has} the meanings given above,
in an inert solvent in the presence of a base with a compound of formula (III)

in which R ^{2 has} the meanings given above and
X ^{1 represents} a suitable leaving group such as chlorine, bromine or iodine,
first to compounds of formula (IV)

in which R ¹ and R ^{2 have} the meanings given above,
reacted, then with intermediate isolation or in a one-pot reaction in the presence of a base to give compounds of the formula (V)

in which R ¹ and R ^{2 have} the meanings given above,
cyclized, then in an inert solvent in the presence of a base with a compound of formula (VI)

in which
X ² and X ^{3 are the} same or different and for a suitable leaving group such as chlorine, bromine or iodine, in compounds of the formula (VII)

in which R ¹ , R ² and X ^{3 have} the meanings given above,
transferred, finally reacted with ammonia with cyclization in an inert solvent and the resulting compounds of the formula (I), if appropriate with the appropriate solvents and / or bases or acids, are converted into their solvates, salts and / or solvates of the salts.

As a solvent for the process step (II) + (III) → (IV) inert organic solvents are suitable, that do not change under the reaction conditions. For this belong Halogenated hydrocarbons such as dichloromethane, trichloromethane, carbon tetrachloride, Trichloroethane, tetrachloroethane, 1,2-dichloroethane or trichlorethylene, Ethers such as diethyl ether, dioxane, tetrahydrofuran, glycol dimethyl ether or diethylene glycol dimethyl ether, hydrocarbons such as benzene, Xylene, toluene, hexane, cyclohexane or petroleum fractions, esters such as ethyl acetate, Ketones such as acetone or 2-butanone, heteroaromatics such as pyridine, amides such as dimethylformamide, dialkyl sulfoxides such as dimethyl sulfoxide, or Nitriles such as acetonitrile. It is also possible to use mixtures of the above solvent use. Dimethylformamide is preferred.

As a base for process step (II) + (III) → (IV) are the usual ones inorganic or organic bases. These include - preferred Alkali or alkaline earth carbonates such as sodium, potassium or calcium carbonate, Alkali hydrides such as sodium hydride, amides such as lithium bis (trimethylsilyl) amide or lithium diisopropylamide, or organic amines such as pyridine, 4-NN-dimethylaminopyridine, 4-pyrrolidinopyridine, triethylamine, ethyldiisopropylamine, N-methylmorpholine, N-methylpiperidine, 1,5-diazabicyclo [4.3.0] non-5-ene (DBN) or 1,8-diazabicyclo [5.4.0] -undec-7-ene (DBU). Particularly preferred is triethylamine.

The base comes in a lot from 1 to 5 mol, preferably in an amount of 1 to 2 mol to 1 mole of the compound of formula (II) used.

The reaction generally takes place in a temperature range of 0 ° C up to + 150 ° C, preferably in a temperature range from + 20 ° C to + 100 ° C. With normal, increased or carried out at reduced pressure (e.g. from 0.5 to 5 bar). Generally one works at normal pressure.

As a solvent for the process step (IV) → (V) are also inert organic solvents, which are under do not change the reaction conditions. These include halogenated hydrocarbons such as dichloromethane, trichloromethane, carbon tetrachloride, trichloroethane, Tetrachloroethane, 1,2-dichloroethane or trichlorethylene, ethers such as Diethyl ether, dioxane, tetrahydrofuran, glycol dimethyl ether or Diethylene glycol dimethyl ether, alcohols such as methanol, ethanol, n-propanol, isopropanol, n-butanol or tert-butanol, hydrocarbons such as benzene, xylene, toluene, Hexane, cyclohexane or petroleum fractions, Ketones such as acetone or 2-butanone, Heteroaromatics such as pyridine, amides such as dimethylformamide, dialkyl sulfoxides such as dimethyl sulfoxide, or nitriles such as acetonitrile. It is the same possible, Mixtures of the solvents mentioned use. Methanol and ethanol are preferred.

Suitable as a base for process step (IV) → (V) the usual inorganic or organic bases. These preferably include alkali or Alkaline earth carbonates such as sodium, potassium or calcium carbonate, alkali hydroxides such as lithium, sodium or potassium hydroxide, alkali alcoholates such as Sodium or potassium methoxide, sodium or potassium ethanolate or Potassium tert-butoxide, alkali hydrides such as sodium hydride, amides such as Lithium bis (trimethylsilyl) amide or lithium diisopropylamide, or organic amines such as pyridine, 4-N, N-dimethylaminopyridine, 4-pyrrolidinopyridine, Triethylamine, ethyldiisopropylamine, N-methylmorpholine, N-methylpiperidine, 1,5-diazabicyclo [4.3.0] non-5-ene (DBN) or 1,8-diazabicyclo [5.4.0] undec-7-ene (DBU). Sodium methoxide and sodium ethoxide are particularly preferred.

The base comes in a lot from 0.5 to 5 mol, preferably in an amount of 1 to 2 mol to 1 mol of the compound of formula (IV) used.

The reaction generally takes place in a temperature range of 0 ° C up to + 120 ° C, preferably in a temperature range from + 20 ° C to + 100 ° C. With normal, increased or carried out at reduced pressure (e.g. from 0.5 to 5 bar). Generally one works at normal pressure.

Suitable solvents for process step (V) + (VI) → (VII) are all inert solvents which do not change under the reaction conditions. These include halogenated hydrocarbons such as Di chloromethane, trichloromethane, tetrachloromethane, trichloroethane, tetrachloroethane, 1,2-dichloroethane or trichlorethylene, ethers such as diethyl ether, dioxane, tetrahydrofuran, glycol dimethyl ether or diethylene glycol dimethyl ether, hydrocarbons such as benzene, xylene, toluene, ether or cyclone such as ethane, cyclone such as ethane, cyclone, hexane, ethane, cyclone, hexane, ethane, cyclone, hexane, ethane, cyclone such as ethane, ether or cyclone, such as ethane, ethane, cyclone, ethane, cyclone, ethane, cyclone, ethane, cyclone, ethane, cyclone, hexane, ethane, ethane, cyclone, ethane, ethane, cyclohexane, ethane, cyclone, ethane, cyclone, ethane, ethane, cyclone, ethane, ethane, cyclone, ethane, ethane, cyclone, ethane, cyclone, ethane, cyclone, ethane, cyclone, ethane, cyclone; Acetone or 2-butanone, amides such as dimethylformamide, dialkyl sulfoxides such as dimethyl sulfoxide, or nitriles such as acetonitrile. It is also possible to use mixtures of the solvents mentioned. Dichloromethane and trichloromethane are preferred.

As a base for process step (V) + (VI) → (VII) are the usual ones inorganic or organic bases. These preferably include alkali or Alkaline earth carbonates and bicarbonates such as sodium, potassium or calcium carbonate and sodium or potassium hydrogen carbonate, alkali hydrides such as sodium hydride, amides such as lithium bis (trimethylsilyl) amide or Lithium diisopropylamide, or organic amines such as pyridine, 4-NN-dimethylaminopyridine, 4-pyrrolidinopyridine, triethylamine, ethyldiisopropylamine, N-methylmorpholine, N-methylpiperidine, 1,5-diazabicyclo [4.3.0] non-5-ene (DBN) or 1,8-diazabicyclo [5.4.0] undec-7-ene (DBU). Sodium bicarbonate is particularly preferred.

The base comes in a lot from 1 to 10 mol, preferably in an amount of 1 to 5 mol to 1 mole of the compound of formula (V) used.

The reaction generally takes place in a temperature range from -20 ° C to + 50 ° C, preferably in a temperature range from -20 ° C up to + 20 ° C. The reaction can be normal, increased or reduced Printing done (e.g. from 0.5 to 5 bar). Generally you work at Normal pressure.

As a solvent for the process step (VII) → (I) all inert solvents are suitable, that do not change under the reaction conditions. For this belong Halogenated hydrocarbons such as dichloromethane, trichloromethane, carbon tetrachloride, Trichloroethane, tetrachloroethane, 1,2-dichloroethane or trichlorethylene, ether such as diethyl ether, dioxane, tetrahydrofuran, glycol dimethyl ether or Diethylene glycol dimethyl ether, or hydrocarbons such as benzene, Xylene, toluene, hexane, cyclohexane or petroleum fractions. It is the same possible mixtures of the solvents mentioned use. Dioxane is preferred.

The compounds of the formulas (II), (III) and (VI) are commercially available, known from the literature or according to the usual literature Methods can be produced [cf. e.g. J. Med. Chem. 13, 674-680 (1970)].

The inventive method can by following formula scheme can be illustrated:

scheme

The compounds of the invention show a unpredictable, valuable pharmacological spectrum of action and are therefore suitable for use as medicinal products for treatment and / or prophylaxis of diseases in humans and animals.

The compounds according to the invention act as antagonists of the P2X ₄ receptor.

Because of their pharmacological properties, the compounds according to the invention can be used alone or in combination with other medicaments for the treatment and / or prophylaxis of inflammatory diseases. In particular, they are suitable for the treatment of chronic inflammatory diseases of the vascular intima such as arteriosclerosis and restenosis, of inflammatory diseases of the central nervous system such as multiple sclerosis and pain, of inflammatory Connective tissue diseases such as rheumatoid arthritis, chronic polyarthritis, panniculitis and tendinitis, ankylosing spondylitis, inflammatory skin diseases such as psoriasis, psoriasis and neurodermatitis, chronic inflammatory bowel diseases such as enteritis, enterocolitis, Crohn's disease and inflammatory diseases of the ulcerative colitis small airways as well as myositis and endocarditis.

The compounds of the invention can be used alone or preferably in combination with other active ingredients if necessary from the group CETP inhibitors, antidiabetics, antioxidants, Thyroid hormones and / or thyroid mimetics, inhibitors of HMG-CoA reductase, HMG-CoA reductase gene expression inhibitors, squalene synthase inhibitors, ACAT inhibitors, cholesterol absorption inhibitors, fibrates, MTP inhibitors, Trigyceride-lowering agent, nicotinic acid and derivatives, antiplatelet agents, anticoagulants, Calcium antagonists, ACE inhibitors, angiotensin II receptor antagonists, Beta blockers and steroidal and non-steroidal anti-phlogistics be administered.

The effectiveness of the compounds according to the invention let yourself check, for example, using the tests described in the example section.

Another subject of the present Invention are medicaments which contain at least one compound according to the invention, preferably together with one or more pharmacologically acceptable auxiliary substances or carriers included, and their use for the aforementioned purposes.

The compounds of the invention can be systemic and / or act locally. To this end, they can do so in an appropriate manner can be applied, e.g. oral, parenteral, pulmonary, nasal, sublingual, lingual, buccal, rectal, dermal, transdermal, conjunctival, otic or as an implant or stent. Oral application is preferred.

For these application routes can the active ingredients are administered in suitable application forms. For the oral application are suitable according to the state of the art, rapid and / or modified application forms delivering the active ingredient, such as. Tablets (non-coated as well as coated tablets, e.g. with enteric coatings), capsules, Dragees, granules, pellets, powders, emulsions, suspensions and Solutions. Parenteral administration can be done by bypassing an absorption step happen (e.g. intravenously, intraarterial, intracardial, intraspinal or intralumbal) or with absorption (e.g. intramuscular, subcutaneous, intracutaneous or intraperitoneal). Suitable for parenteral administration themselves as application forms Injection and infusion preparations in the form of solutions, Suspensions, emulsions, lyophilisates and sterile powders.

For the other application routes are e.g. Inhalation dosage forms (e.g. powder inhalers, nebulizers), nose drops / solutions, Sprays, lingual, sublingual or buccal tablets or capsules, suppositories, ear and eye preparations, vaginal capsules, aqueous Suspensions (lotions, shake mixes), lipophilic suspensions, ointments, creams, milk, pastes, powder, Implants or stents.

The active ingredients can be in a manner known per se in the listed Application forms are transferred. This is done using inert non-toxic, pharmaceutical suitable auxiliaries. Which includes et al excipients (e.g. microcrystalline cellulose), solvents (e.g. liquid polyethylene glycols), emulsifiers (e.g. sodium dodecyl sulfate), dispersing agents (e.g. polyvinylpyrrolidone), synthetic and / or natural Biopolymers (e.g. albumin), stabilizers (e.g. antioxidants such as. Ascorbic acid), Colorants (e.g. inorganic pigments such as iron oxides) or taste and / or odor correctives.

In general, it has proven to be beneficial proven in parenteral application amounts from about 0.001 to 10 mg / kg, preferably about 0.005 to 3 mg / kg body weight to achieve more effective Deliver results. In the case of oral application, the amount is about 0.001 to 100 mg / kg, preferably about 0.005 to 30 mg / kg body weight.

Nevertheless, it may be necessary be different from the amounts mentioned, depending on of body weight, Application route, individual behavior towards the active ingredient, on the preparation and the time or interval at which the application takes place. In some cases it can be sufficient with less than the aforementioned minimum amount get along while in other cases the specified upper limit was exceeded must become. In the case of application of larger quantities, it can be recommended be spread over the day in several separate doses.

The percentages in the following Unless otherwise stated, tests and examples are percentages by weight; Parts are parts by weight. Solvent ratios, dilution ratios and obtain concentration information from liquid / liquid solutions each on the volume.

The following examples explain The invention. The invention is not limited to the examples.

Abbreviations CI chemical ionization (for MS) DC TLC DCI direct chemical ionization (for MS) DMF N, N-dimethylformamide DMSO dimethyl sulfoxide theory the theory (with yield) IT I Electrospray ionization (for MS) LC / MS Liquid chromatography-coupled mass spectroscopy MS mass spectroscopy NMR Nuclear Magnetic Resonance Spectroscopy R _f Retention index (for DC) R _t Retention time (for LC / MS)

LC / MS methods

Method 1

Device type MS: Micromass ZQ; Device type HPLC: Waters Alliance 2790; Pillar: Uptisphere C 18, 50 mm × 2.0 mm, 3.0 µm; Eluent B: acetonitrile + 0.05% formic acid, eluent A: water + 0.05% formic acid; Gradient: 0.0 min 5% B → 2.0 min 40% B → 4.5 min 90% B → 5.5 min 90% B; Oven: 45 ° C; Flow: 0.0 min 0.75 ml / min → 4.5 min 0.75 ml / min → 5.5 min 1.25 ml / min; UV detection: 210 nm.

Method 2

Instrument: Micromass Platform LCZ with HPLC Agilent series 1100; Pillar: Grom-SIL 120 ODS-4 HE, 50 mm × 2.0 mm, 3 µm; Eluent A: 1 l water + 1 ml 50% formic acid, eluent B: 1 l acetonitrile + 1 ml of 50% formic acid; Gradient: 0.0 min 100% A → 0.2 min 100% A → 2.9 min 30% A → 3.1 min 10% A → 4.5 min 10% A; Oven: 55 ° C; Flow: 0.8 ml / min; UV detection: 208-400 nm.

A. Starting compounds

Example I (3-aminobenzofuran-2-yl) - (3-bromophenyl) methanone

2.00 g (16.8 mmol) of 2-hydroxybenzonitrile, 4.67 g (16.8 mmol) of 3-bromophenacyl bromide and 1.87 g (18.5 mmol) of triethylamine in 20 ml of dimethylformamide are stirred at 70 ° C. for 2 h. After adding 100 ml of ethyl acetate, the reaction mixture is washed with water (3 × 100 ml) and saturated sodium chloride solution (2 × 100 ml). The organic phase is dried over magnesium sulfate and the solvent is removed under reduced pressure. The residue is dissolved in 30 ml of ethanol. After adding 5.98 g (18.5 mmol) of sodium ethanolate, the mixture is heated under reflux for 2 h. 50 ml of ethyl acetate are added and the mixture is washed with water (3 × 100 ml) and saturated sodium chloride solution (2 × 100 ml). The organic phase is dried over magnesium sulfate. After removal of the solvent under reduced pressure, 5.08 g (92% of theory) of the desired product are obtained.
MS (DCI): m / z = 315.9 [M + H] ^+
1 H NMR (200 MHz, DMSO-d ₆ ): δ = 7.27-7.36 (m, 1H); 7.49-7.65 (m, 5H); 7.77-7.84 (m, 1H); 8.04-8.20 (m, 3H).

Example II 2-Bromo-N- {2 - [(3-bromophenyl) carbonyl] benzofuran-3-yl} acetamide

3.53 g (17.5 mmol) of bromoacetyl bromide are added at 0 ° C. to a mixture of 5.03 g (15.9 mmol) of the compound from Example I and 5.35 g (63.6 mmol) of sodium hydrogen carbonate in 250 ml of chloroform. The mixture is stirred at 0 ° C for 1 h. After removing the solvent under reduced pressure, 200 ml of ethyl acetate is added, and the mixture is washed with saturated sodium hydrogen carbonate solution (100 ml) and saturated sodium chloride solution (100 ml). The organic phase is dried over magnesium sulfate. After removal of the solvent under reduced pressure, 5.81 g (83% of theory) of the desired product are obtained.
MS (CI): m / z = 436 [M + H] ^+
1 H NMR (200 MHz, DMSO-d ₆ ): δ = 4.20 (s, 2H); 7.37-8.14 (m, 8H); 10.98 (s, 1H).

B. embodiments

Example 1 5- (3-Bromophenyl) -1,3-dihydro-2H-benzofuro [3,2-e] -1,4-diazepin-2-one

245 ml (123 mmol) of a 0.5 M solution of ammonia in dioxane are added to 5.37 g (12.3 mmol) of the compound from Example II in 100 ml of diethyl ether. The mixture is stirred for 2 days at room temperature. 100 ml of ethyl acetate are added and the mixture is washed with water (3 × 250 ml) and saturated sodium chloride solution. The organic phase is dried over magnesium sulfate and the solvent is removed under reduced pressure. 1.81 g (42% of theory) of the desired product are obtained.
R _f = 0.24 (dichloromethane / methanol 100: 2)
MS (ESI): m / z = 355 [M + H] ⁺
LC / MS (method 1): R _t = 3.47 min., M / z = 354 [M] ^+
1 H NMR (200 MHz, DMSO-d ₆ ): δ = 4.45 (s, 2H); 7.37-7.80 (m, 6H); 7.88-7.93 (m, 1H); 7.96-8.03 (m, 1H); 11.59 (s, 1H).

LC/MS (Methode 2): R_t = 3.5 min., m/z = 321 [M + H]⁺. Beispiel 3 9-Brom-l,3-dihydro-5-phenyl-2H-benzofuro[3,2-e]-1,4-diazepin-2-on

LC/MS (Methode 2): R_t = 3.5 min., m/z = 355 [M + H]⁺.The following are obtained in an analogous manner: Example 2 1,3-Dihydro-5- (3-nitrophenyl) -2H-benzofuro [3,2-e] -1,4-diazepin-2-one

LC / MS (Method 2): R _t = 3.5 min., M / z = 321 [M + H] ⁺ . Example 3 9-bromo-1,3-dihydro-5-phenyl-2H-benzofuro [3,2-e] -1,4-diazepin-2-one

LC / MS (Method 2): R _t = 3.5 min., M / z = 355 [M + H] ⁺ .

C. Description of the biological Testing:

a) Cellular assay

The P2X ₄ receptor is a ligand-activated ion channel. The binding of the agonist ATP leads to the activation of the P2X ₄ receptor, opening of the ion channel and influx of extracellular calcium into the cell. This calcium influx is measured using the calcium-sensitive photoprotein aequorin. For this purpose, a recombinant CHO cell line (Chinese hamster ovary cells) with constitutive expression of the human P2X ₄ receptor and of apoaequorin was produced.

To carry out the experiment, the CHO cells are sown 1-2 days beforehand on microtiter plates in 96, 384 or 1536-well format, in accordance with the microtiter plate format used with 5000 (96 format), 2000 (384 format) or 500 (1536 format) cells per well. On the day of the experiment, the cell culture medium is removed and the cells are incubated for 4 hours in physiological saline (Tyrode buffer) with 5 μg / ml coelenterazine. Test substances are added 5 minutes before the actual experiment. To activate the P2X ₄ receptor, ATP is then added in a concentration of 1-2 μM and the ATP-induced calcium signal is measured in a luminometer as aequorin luminescence.

Substances with antagonistic activity at the P2X ₄ receptor can inhibit the ATP-induced calcium signal either by interference with the binding of ATP to the P2X ₄ receptor, by preventing the opening of the channel or by blocking the calcium influx through the opened channel ,

In this test, the exemplary embodiments 1-3 show IC ₅₀ values of 0.5, 2 and 0.6 μM.

b) ATP-induced oxygen radical formation (ROS) in primary human monocytes:

The assay is performed in Hank's Balanced Salt Solution (HBSS) to which 10 mM glucose is added. Monocytes are isolated, for example using the "Becton Dickinson Vacutainer System" as described by the manufacturer, and suspended in HBSS. In principle, the detection of oxygen radicals is carried out after the luminol-enhanced chemiluminescence method in the presence of horse radish peroxidase (HRPO) [H. Lundqvist, C. Dahlgren, Free Radic. Biol. Med. 20 (6): 785-92 (1996)].

First, the substance to be investigated, luminol (50 μM final concentration), HRPO (10 U / ml final concentration) are incubated with 5 × 10 ⁵ monocytes for 15 min at 37 ° C. Then ATP (100 μM final concentration) is added to the test mixture. The final volume in the test mixture is 200 μl. Immediately after adding ATP, the ROS formation is monitored with a microplate luminometer over a period of 120 seconds.

c) ATP-induced chemotaxis of primary human monocytes:

Monocytes are isolated from blood using standard methods. The chemotaxis of the monocytes is observed in a Transwell system [CC Bleul et al., J. Exp. Med. 184: 1101-1109 (1996)]. The membrane used (3 μm pore size, polyethylene terephthalate, Falcon) is first coated with fibronectin. 10 ⁵ monocytes in RPMI 1640 medium are added to the upper chamber. The lower chamber contains varying concentrations of stimulus or constant stimulus concentration (500 μM ATP or 10 nM MCP-1) and varying concentrations of the test substance. The substances to be examined are in both chambers. The test mixture is incubated for 3 h at 37 ° C. and 5% CO ₂ [W. Falk et al., J. Immunol. Methods 38: 239-247 (1980)]. After the incubation, the cells that have migrated to the lower chamber are determined.

D. Examples for pharmaceutical compositions

The compounds of the invention can be pharmaceutical as follows Preparations are transferred:

tablet

composition

100 mg of the compound of example 1, 50 mg lactose (monohydrate), 50 mg corn starch (native), 10 mg polyvinylpyrrolidone (PVP 25) and 2 mg magnesium stearate. Tablet weight 212 mg. diameter 8 mm, radius of curvature 12 mm.

manufacturing

The mixture of active ingredient, lactose and strength comes with a 5% solution (m / m) of the PVP granulated in water. The granulate is after Dry with the magnesium stearate for 5 min. long mixed. This mix comes with a usual Tablet press pressed (for tablet format see above). As Guide value for the pressing uses a pressing force of 15 kN.

Oral suspension

composition

1000 of the compound of Example 1, 1000 mg of ethanol mg (96%), 400 mg of Rhodigel ^® (xanthan gum of the Fa. FMC, Pennsylvania, USA) and 99 g of water.

A single dose of 100 mg compound of the invention correspond to 10 ml oral suspension.

manufacturing

The Rhodigel is suspended in ethanol the active ingredient is added to the suspension. The mixture is added with stirring Water. Until the swelling of the Rhodigel is completed, approx. 6 h stirred.

Claims (10)

Compounds of the general formula (I)

in which R ^{1 is} halogen and R ^{2 is} hydrogen, halogen, nitro, cyano or a group of the formula -C (O) -OR ³ , -C (O) -NR ⁴ R ⁵ , -SO ₂ -OR ³ or -SO ₂ -NR ⁴ R ⁵ , wherein R ³ , R ⁴ and R ⁵ independently of one another are hydrogen or (C ₁ -C ₆ ) alkyl, or R ^{1 is} hydrogen and R ^{2 is} halogen, nitro, cyano or a group of the formula -C (O) -OR ³ , -C (O) -NR ⁴ R ⁵ , -SO ₂ -OR ³ or -SO ₂ -NR ⁴ R ⁵ , wherein R ³ , R ⁴ and R ^{5 are} independently hydrogen or (C ₁ -C ₆ ) alkyl, means, and their salts, solvates and solvates of the salts. Compounds of the general formula (I) according to Claim 1, in which R ^{1 is} chlorine or bromine and R ^{2 is} hydrogen, chlorine, bromine, nitro, cyano or a group of the formula -C (O) -OR ³ or -C (O) - NR ⁴ R ⁵ , in which R ³ , R ⁴ and R ⁵ independently represent hydrogen or (C ₁ -C ₄ ) alkyl, or R ^{1 is} hydrogen and R ^{2 is} chlorine, bromine, nitro, cyano or a group of the formula - C (O) -OR ³ or -C (O) -NR ⁴ R ⁵ , where R ³ , R ⁴ and R ⁵ independently of one another are hydrogen or (C ₁ -C ₄ ) alkyl. Compounds of formula (Ia)

in which R ^{1 is} chlorine or bromine and R ^{2 is} hydrogen, chlorine, bromine, nitro or cyano, or R ^{1 is} hydrogen and R ^{2 is} chlorine, bromine, nitro or cyano, and their salts, solvates and solvates of the salts. Process for the preparation of compounds of the formula (I) or (Ia) as defined in claims 1 to 3, characterized in that compounds of the formula (II)

in which R ^{1 has} the meanings given above, in an inert solvent in the presence of a base with a compound of the formula (III)

in which R ^{2 has} the meanings given above and X ^{1 represents} a suitable leaving group such as chlorine, bromine or iodine, first to give compounds of the formula (N)

in which R ¹ and R ^{2 have} the meanings given above, then reacting them with intermediate isolation or in a one-pot reaction in the presence of a base to give compounds of the formula (V)

in which R ¹ and R ^{2 have} the meanings given above, cyclized, then in an inert solvent in the presence of a base with a compound of the formula (VI)

in which X ² and X ^{3 are the} same or different and represent a suitable leaving group such as chlorine, bromine or iodine, in compounds of the formula (VII)

in which R ¹ , R ² and X ^{3 have} the meanings given above, converted, finally reacted with ammonia with cyclization in an inert solvent and the resulting compounds of the formula (I), if appropriate, with the corresponding solvents and / or bases or acids in their Solvates, salts and / or solvates of the salts are transferred. Compounds according to one of claims 1 to 3 for treatment and / or prophylaxis of diseases. Use of compounds according to one of claims 1 to 3 for the manufacture of pharmaceuticals. Medicaments containing at least one of the compounds according to one of the claims 1 to 3 in combination with at least one pharmaceutically acceptable pharmaceutically acceptable carrier or excipients. Use of compounds according to one of claims 1 to 3 for the manufacture of a medicament for treatment and / or prophylaxis of arteriosclerosis and restenosis. Medicament according to claim 7 for treatment and / or Prophylaxis of arteriosclerosis and restenosis. Method of combating arteriosclerosis and restenosis in humans and animals by administering an effective one Amount of at least one compound according to one of claims 1 to Third