DE102006031314A1 - Use of 1,4-diaryl-dihydropyrimidin-2-one derivative for producing a medicament for the treatment and/or prophylaxis of e.g. pulmonary arterial hypertension, chronic-obstructive lung diseases and sleep apnea syndrome - Google Patents

Abstract

Use of 1,4-diaryl-dihydropyrimidin-2-one derivative (I) or its salts, solvates or solvates of the salts, for producing a medicament for the treatment and/or prophylaxis of pulmonary arterial hypertension. Use of 1,4-diaryl-dihydropyrimidin-2-one derivative of formula (I) or its salts, solvates or solvates of the salts, for producing a medicament for the treatment and/or prophylaxis of the pulmonary arterial hypertension. R 1>H, -(CH 2) nC(=O)-O-R 5> or benzoic acid of formula (a) or (b); R 5>H or 1-4C-alkyl; R 2>CN or -C(=O)-R 6> or -C(=O)-O-R 6>; R 6>1-6C-alkyl or 3-6C-cycloalkyl (which is partially or substituted two times with hydroxy, 1-4C-alkoxy, hydroxycarbonyl, amino, mono- and/or di-1-4C-alkylamino, where respectively, CH 2 is replaced with an O-atom); R 3>, R 4>H, F or Cl; X : CH or N; and asterisk : connecting place with the N-atom. Independent claims are included for: (1) a combination comprising (I) and an active substance from kinase-inhibitor, stimulator and activator of the soluble guanylate cyclase, prostacyclin-analogues, endothelium receptor-antagonist and phosphodiesterase-inhibitors; (2) a medicament comprising the combination; and (3) method for the treatment and/or prophylaxis of the pulmonary arterial hypertension with humans and animals by administering (I), the combination or the medicament in combination with inert, non-toxic auxiliary materials. [Image] ACTIVITY : Hypotensive; Respiratory-Gen; Vulnerary; Respiratory-gen.; CNS-Gen; Thrombolytic; Antiinflammatory. MECHANISM OF ACTION : Neutrophile elastase inhibitor. The neutrophile elastase inhibitory activity of (I) was tested using human neutrophile elastase. The result showed that (I) exhibited a median inhibitory concentration (IC 5 0) of 14.8 nM.

Description

The The present application relates to the use of 1,4-diaryl-dihydropyrimidin-2-one derivatives of the formula (I) for the treatment and / or prophylaxis of pulmonary arterial Hypertension and its use for the manufacture of a drug for the treatment and / or prophylaxis of pulmonary arterial hypertension.

Pulmonary arterial hypertension (PAH) is a progressive lung disease that, if untreated, leads to an average death within 2.8 years of diagnosis. An increasing constriction of the pulmonary circulation leads to an increase in the burden on the right heart, which can lead to right heart failure. By definition, chronic pulmonary hypertension has a pulmonary arterial mean pressure (mPAP) of> 25 mmHg at rest or> 30 mmHg under exercise (normal value <20 mmHg). The pathophysiology of pulmonary arterial hypertension is characterized by vasoconstriction and remodeling of the pulmonary vessels. In chronic PAH, the vascular musculature increases in size, followed by a slow remodeling of the musculature to connective tissue. This increasing obliteration of the pulmonary circulation leads to a progressive load on the right heart, which leads to a reduced output of the right heart and ultimately ends in right heart failure. With a prevalence of 1-2 per million, PAH is an extremely rare disease ( GE D'Alonzo et al., Ann. Intern. Med. 1991, 115, 343-349 ). The mean age of the patients was estimated at 36 years, only 10% of the patents were over 60 years old (ibid). Significantly more women than men are affected.

Despite all advances in the treatment of pulmonary arterial hypertension, there is no prospect of curing this serious disease. Standard therapies on the market (eg, prostacyclin analogs, endothelin receptor antagonists, phosphodiesterase inhibitors) are able to improve the quality of life, exercise tolerance, and prognosis of patients. However, the applicability of these drugs is limited by the partly serious side effects and / or complex application forms. The period of time during which a patient's clinical situation can be improved or stabilized under specific monotherapy is limited. Finally, there is a therapy escalation and thus a combination therapy in which several drugs have to be given at the same time. New combination therapies are one of the most promising future treatment options for the treatment of pulmonary arterial hypertension ( Ghofrani et al., Herz 2005, 30, 296-302 ). In this context, the exploration of new pharmacological mechanisms for the treatment of PAH is of particular interest. New therapies should be combinable with the known com. To minimize the risk of disruptive drug-drug interactions in such a combination therapy, these new drugs should not or only to a very limited extent inhibit metabolising P450 CYP enzymes.

Of the Term "pulmonary arterial hypertension " certain forms of pulmonary hypertension, e.g. of the World Health Organization (WHO) (Clinical Classification of Pulmonary Hypertension, Venice 2003).

The Pulmonary arterial hypertension involves the idiopathic pulmonary Arterial hypertension (IPAH, formerly also as primary pulmonary hypertension), familial pulmonary arterial hypertension (FPAH) and Associated Pulmonary Arterial Hypertension (APAH), which is associated with collagenosis, congenital systemic-pulmonary Shuntvitien, portal hypertension, HIV infections, ingestion certain drugs and medicines, with other diseases (thyroid diseases, Glycogen storage disorders, Gaucher disease, hereditary telangiectasia, hemoglobinopathies myeloproliferative disorders, splenectomy), with diseases with a significant venous / capillary Involvement such as pulmonary veno-occlusive disease and pulmonary capillary hemangiomatosis, as well as the persistent pulmonary hypertension of newborns.

Human leukocyte elastase (HLE, EC 3.4.21.37), also called human neutrophil elastase (HNE, hNE), belongs to the family of serine proteases. The proteolytic enzyme is found in the azurophilic granules of polymorphonuclear leukocytes (polymorphonuclear leukocytes, PMN leukocytes). Intracellular elastase plays an important role in the defense against pathogens by breaking down foreign particles taken up via phagocytosis. Activated neutrophils release the HNE from the granules into the extracellular space (extracellular HNE), leaving some of the released HNE on the outside of the neutrophil cell membrane (membrane-bound HNE). The highly active enzyme is able to break down a variety of connective tissue proteins, such as the proteins elastin, collagen and fibronectin. Elastin is found in high concentrations in all tissue types that show high elasticity, eg in the lungs and in arteries. In a variety of pathological processes (eg tissue injury) the HNE plays a role Le tissue removal and remodeling (tissue remodeling). In addition, HNE is an important modulator in inflammatory processes. For example, HNE induces increased gene expression of interleukin-8 (IL-8). It is therefore believed that the HNE is present in many diseases of the lung (eg, chronic obstructive pulmonary disease, COPD, acute respiratory distress syndrome, ARDS, cystic fibrosis, CF) Emphysema) and also in diseases of the cardiovascular system (eg tissue changes after a myocardial infarction and in heart failure) plays an important role.

In animal models and in patients with increased pulmonary arterial pressure (pulmonary arterial hypertension), connective tissue fragmentation (internal elastic lamina) has been observed [ Rabinovitch et al., Lab. Invest. 55, 632-653 (1986) ]. In animal models of pulmonary arterial hypertension (hypoxic and monocrotaline rat model) increased elastase activity was shown, which was associated with fragmentation of the connective tissue [ Todorovich-Hunter et al., Am. Rev. Respir. Dis. 146, 213-223 (1992) ]. It is thought that the tissue remodeling to be observed is induced in the course of disease progression of pulmonary arterial hypertension by elastase-mediated release of connective tissue growth factors, eg basic fibroblast growth factor (bFGF). Rabinovitch, Am. J. Physiol. 277, L5-L12 (1999) ]. In the hypoxic mouse model for pulmonary arterial hypertension, a positive effect with an overexpressed elastase inhibitor protein could be shown [ Zaidi et al., Circulation 105, 516-521 (2002) ]. In the monocrotaline rat model for pulmonary arterial hypertension, a positive effect could be demonstrated with synthetic, low molecular weight elastase inhibitors; this also had a favorable effect on tissue remodeling [ Cowan et al., Nature Med. 6, 698-702 (2000) ]. However, all previously known low molecular weight elastase inhibitors are less selective, chemically reactive and / or only orally available, which has hitherto thwarted the clinical development of an oral elastase inhibitor in these indications.

In general, it is believed that elastase-mediated pathological processes underlie a shifted balance between free elastase and the body's elastase inhibitor protein (mainly alpha-1 antitrypsin, AAT) [ Stockley, Neutrophils and protease / antiprotease imbalance, Am. J. Respir. Crit. Care Med. 160, 49-52 (1999) ]. AAT is present in plasma in high excess and thus very quickly neutralizes free HNE. In various pathological processes, the concentration of free elastase is increased, so that locally the balance between protease and protease inhibitor is shifted in favor of the protease. In addition, the membrane-bound elastase of the activated PMN cells is largely protected from inhibition by AAT. The same applies to the free elastase, which is located in a difficult-to-access microcompartment between the neutrophilic cell and the adjacent tissue cell (eg endothelial cell). In addition, strongly oxidizing conditions prevail in the environment of activated leukocytes (English: oxidative burst), whereby AAT is oxidized and loses several orders of magnitude in the inhibitory effect.

New Elastase-inhibiting agents (exogenously administered inhibitors the HNE) should therefore have a low molecular weight, in order to be able to even the membranous HNE and in the protected microcompartment reach and inhibit HNE (see above). This is also a good stability the substances in vivo necessary (low in vivo clearance). In addition, should These compounds are stable under oxidative conditions not to lose inhibitory potency in the disease process. In indications where combination therapies are performed or for the future to be expected, such as the PAH, is particular a low level of interaction with enzymes beneficial to the drug agents could change and dismantle (P450 CYP enzymes).

In WO 2004/024700 . WO 2004/024701 . WO 2005/082863 and WO 2005/082864 Various 1,4-diaryl-dihydropyrimidin-2-one derivatives are disclosed as HNE inhibitors for the treatment of chronic obstructive pulmonary disease, acute coronary syndrome, myocardial infarction and heart failure.

Surprisingly It has now been found that certain 1,4-diaryl-dihydropyrimidin-2-one derivatives in particular Way for the treatment of pulmonary arterial hypertension (PAH). These compounds described below are low molecular weight, non-reactive, selective and potent inhibitors of neutrophils Elastase that over a sufficiently high bioavailability after oral administration and / or good solubility for parenteral administration feature as well as low in vitro clearance to hepatocytes and one only have low inhibition of CYP enzymes from microsomes. they provide thus promising starting points for new drugs for treatment pulmonary arterial hypertension as a single therapy or in combination with other active substances.

in welcher
X für CH oder N steht,
R¹ für Wasserstoff, eine Gruppe der Formel -(CH₂)_n-C(=O)-O-R^1A oder eine Gruppe der Formel

steht, worin
* die Verknüpfungsstelle mit dem N-Atom bedeutet,
n die Zahl 1 oder 2 bedeutet
und
R^1A Wasserstoff oder (C₁-C₄)-Alkyl bedeutet,
R² für Cyano oder eine Gruppe der Formel -C(=O)-R^2A oder -C(=O)-O-R^2A steht, worin
R^2A (C₁-C₆)-Alkyl oder (C₃-C₆)-Cycloalkyl bedeutet, welche ihrerseits bis zu zweifach, gleich oder verschieden, mit Hydroxy, (C₁-C₄)-Alkoxy, Hydroxycarbonyl, Amino, Mono- und/oder Di-(C₁-C₄)-alkylamino substituiert sein können und worin jeweils eine CH₂-Gruppe, soweit in einer chemisch stabilen Verbindung resultierend, gegen ein O-Atom ausgetauscht sein kann,
und
R³ entweder für Wasserstoff und
R⁴ für Wasserstoff, Fluor oder Chlor steht oder
R³ für Fluor oder Chlor und
R⁴ für Wasserstoff steht,
sowie ihrer Salze, Solvate und Solvate der Salze zur Herstellung eines Arzneimittels zur Behandlung und/oder Prophylaxe der pulmonalen arteriellen Hypertonie.The present invention relates to the use of compounds of the general formula (I)

in which
X is CH or N,
R ^{1 is} hydrogen, a group of the formula - (CH ₂ ) _n -C (= O) -OR ^1A or a group of the formula

stands in which
* means the point of attachment to the N-atom,
n is the number 1 or 2
and
R ^{1A is} hydrogen or (C ₁ -C ₄ ) -alkyl,
R ^{2 is} cyano or a group of the formula -C (= O) -R ^2A or -C (= O) -OR ^2A in which
R ^{2A is} (C ₁ -C ₆ ) -alkyl or (C ₃ -C ₆ ) -cycloalkyl, which in turn is up to twice, identically or differently, with hydroxy, (C ₁ -C ₄ ) -alkoxy, hydroxycarbonyl, amino, Mono- and / or di- (C ₁ -C ₄ ) -alkylamino may be substituted and in which in each case a CH ₂ group, as far as resulting in a chemically stable compound, may be exchanged for an O atom,
and
R ³ either for hydrogen and
R ^{4 is} hydrogen, fluorine or chlorine or
R ^{3 is} fluorine or chlorine and
R ^{4 is} hydrogen,
and their salts, solvates and solvates of the salts for the manufacture of a medicament for the treatment and / or prophylaxis of pulmonary arterial hypertension.

steht, worin
* die Verknüpfungsstelle mit dem N-Atom bedeutet,
R² für Cyano, Acetyl, Cyclobutylcarbonyl, Methoxycarbonyl, Ethoxycarbonyl oder 2-Hydroxyethoxycarbonyl steht,
R³ für Wasserstoff steht
und
R⁴ für Wasserstoff oder Fluor steht,
sowie ihrer Salze, Solvate und Solvate der Salze.Preference is given here to the use of compounds of the formula (1) in which
X is CH or N,
R ^{1 is} hydrogen, a group of the formula -CH ₂ -C (= O) -OH or a group of the formula

stands in which
* means the point of attachment to the N-atom,
R ^{2 is} cyano, acetyl, cyclobutylcarbonyl, methoxycarbonyl, ethoxycarbonyl or 2-hydroxyethoxycarbonyl,
R ^{3 is} hydrogen
and
R ^{4 is} hydrogen or fluorine,
and their salts, solvates and solvates of the salts.

und ihrer Salze, Solvate und Solvate der Salze.Particularly preferred here is the use of compounds of the formula (I) having the following structures:

and their salts, solvates and solvates of salts.

Usable according to the invention Compounds, hereinafter also referred to as compounds of the invention are the compounds of formula (I) and their salts, Solvates and solvates of the salts, the compounds encompassed by formula (I) the above or below Formulas and their salts, solvates and solvates of the salts and the of formula (1), hereinafter referred to as exemplary embodiments compounds and their salts, solvates and solvates of the salts, as far as they are concerned the compounds of formula (I), mentioned below not already salts, solvates and solvates of the salts.

The Compounds of the invention can dependent on of their structure in stereoisomeric forms (enantiomers, diastereomers) exist. The invention therefore includes the enantiomers or Diastereomers and their respective mixtures. From such mixtures enantiomers and / or diastereomers can be the stereoisomer isolate uniform components in a known manner.

Provided the compounds of the invention can occur in tautomeric forms, For example, the present invention encompasses all tautomeric forms.

When Salts are physiologically acceptable in the context of the present invention Salts of the compounds of the invention prefers. Also included are salts that are suitable for pharmaceutical applications themselves are not suitable, but for example for insulation or cleaning the compounds of the invention can be used.

physiological Safe salts of the compounds of the invention include acid addition salts of mineral acids, carboxylic acids and sulfonic acids, e.g. Salts of hydrochloric acid, hydrobromic, Sulfuric acid, Phosphoric acid, methane, ethanesulfonic, toluene sulfonic acid, benzenesulfonic, naphthalenedisulfonic, Acetic acid, Trifluoroacetic acid, propionic acid, Lactic acid, Tartaric acid, malic acid, citric acid, fumaric acid, maleic acid and Benzoic acid.

physiological acceptable salts of the compounds of the invention also include Salts more usual Bases, such as by way of example and preferably alkali metal salts (e.g. Sodium and potassium salts), alkaline earth salts (e.g., calcium and magnesium salts) and ammonium salts derived from ammonia or organic amines with 1 to 16 carbon atoms, such as by way of example and preferably ethylamine, Diethylamine, triethylamine, ethyldiisopropylamine, monoethanolamine, Diethanolamine, triethanolamine, dicyclohexylamine, dimethylaminoethanol, procaine, Dibenzylamine, N-methylmorpholine, arginine, lysine, ethylenediamine and N-methylpiperidine.

When In the context of the invention, solvates are those forms of the compounds according to the invention, which in solid or liquid Condition by coordination with solvent molecules a complex form. Hydrates are a special form of solvates in which the coordination with water takes place. As solvates are in the frame Hydrate is preferred in the present invention.

It also includes the present invention also prodrugs of the compounds of the invention. Of the Term "prodrugs" includes compounds, which may themselves be biologically active or inactive, but during their Residence time in the body to compounds of the invention be implemented (for example, metabolically or hydrolytically).

Unless otherwise specified, in the context of the present invention, the substituents have the following meaning:
(C ₁ -C ₆ ) -Alkyl and (C ₁ -C ₄ ) -alkyl are in the context of the invention a straight-chain or branched alkyl radical having 1 to 6 or 1 to 4 carbon atoms. Preferred is a straight-chain or branched alkyl radical having 1 to 4 carbon atoms. Examples which may be mentioned are: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, 1-ethylpropyl, n-pentyl and n-hexyl.

In the context of the invention, (C ₃ -C ₆ ) -cycloalkyl is a monocyclic, saturated cycloalkyl group having 3 to 6 carbon atoms. By way of example and preferably mention may be made of: cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

(C ₁ -C ₄ ) -Alkoxy in the context of the invention is a straight-chain or branched alkoxy radical having 1 to 4 carbon atoms. Examples which may be mentioned by way of example include: methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy and tert-butoxy.

Mono- (C ₁ -C ₄ ) -alkylamino in the context of the invention represents an amino group having a straight-chain or branched alkyl substituent which has 1 to 4 carbon atoms. Examples which may be mentioned by way of example include methylamino, ethylamino, n-propylamino, isopropylamino, n-butylamino and tert-butylamino.

Di (C ₁ -C ₄ ) -alkylamino in the context of the invention is an amino group having two identical or different straight-chain or branched alkyl substituents, each having 1 to 4 carbon atoms. Examples which may be mentioned by way of example and with preference: N, N-dimethylamino, N, N-diethylamino, N-ethyl-N-methylamino, N-methyl-Nn-propylamino, N-isopropyl-N-methylamino, N-isopropyl-Nn-propylamino, N, N-diisopropylamino, Nn-butyl-N-methylamino and N-tert-butyl-N-methylamino.

in welcher X die oben angegebene Bedeutung hat,
in Gegenwart einer Säure oder eines Säureanhydrids in einer 3-Komponenten-Eintopf-Reaktion oder sequentiell mit einer Verbindung der Formel (III)

in welcher R² die oben angegebene Bedeutung hat,
und einer Verbindung der Formel (IV)

in welcher R³ und R⁴ die oben angegebenen Bedeutungen haben,
zu einer Verbindung der Formel (I-A)

in welcher R², R³, R⁴ und X jeweils die oben angegebenen Bedeutungen haben,
umsetzt und diese im Falle, dass R¹ nicht für Wasserstoff steht, in Gegenwart einer Base mit einer Verbindung der Formel (V) R^1*-Z (V),in welcher
R^1* die oben angegebene Bedeutung von R¹ hat, jedoch nicht für Wasserstoff steht,
und
Z für eine Abgangsgruppe wie beispielsweise Halogen, Mesylat, Tosylat oder Triflat steht, reagiert
und die so erhaltenen Verbindungen der Formel (I-A) bzw. (I) nach dem Fachmann bekannten Methoden in ihre Enantiomere und/oder Diastereomere trennt und gegebenenfalls mit den entsprechenden (i) Lösungsmitteln und/oder (ii) Basen oder Säuren in ihre Solvate, Salze und/oder Solvate der Salze überführt.The compounds of the formula (1) which can be used according to the invention are for the most part of WO 2004/024700 . WO 2005/082863 and WO 2005/082864 known. They can be described as described in detail there or in analogy to this. In a general way, the compounds of the formula (I) can be prepared by reacting a compound of the formula (II)

in which X has the meaning given above,
in the presence of an acid or an acid anhydride in a 3-component one-pot reaction or sequentially with a compound of the formula (III)

in which R ^{2 has} the meaning given above,
and a compound of formula (IV)

in which R ³ and R ^{4 have} the meanings given above,
to a compound of the formula (IA)

in which R ² , R ³ , R ⁴ and X are each as defined above,
and in the event that R ¹ does not represent hydrogen, in the presence of a base with a compound of formula (V) R ^{1 *} -Z (V), in which
R ^{1 * has} the abovementioned meaning of R ¹ , but does not stand for hydrogen,
and
Z represents a leaving group such as halogen, mesylate, tosylate or triflate
and the compounds of the formula (IA) or (I) thus obtained are separated by methods known to those skilled in the art into their enantiomers and / or diastereomers and, if appropriate, with the appropriate (i) solvents and / or (ii) bases or acids in their solvates, Salts and / or solvates of the salts converted.

For the process step (II) + (III) + (IV) → (I-A) suitable solvents are usual organic solvents, which do not change under the reaction conditions. To counting For example, ethers such as diethyl ether, diisopropyl ether, methyl tert-butyl ether, 1,2-dimethoxyethane, dioxane or tetrahydrofuran, alcohols such as methanol, Ethanol, n-propanol, isopropanol, n-butanol or tert-butanol, hydrocarbons such as pentane, hexane, cyclohexane, benzene, toluene or xylene, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, trichloromethane or chlorobenzene, or other solvents such as ethyl acetate, acetonitrile, dimethyl sulfoxide or N, N-dimethylformamide. It is also possible Mixtures of the solvents mentioned use. Preference is given to using tetrahydrofuran or dioxane.

When Acid for the process step (II) + (III) + (IV) → (I-A) are usual inorganic or organic acids or acid anhydrides. These include preferably carboxylic acids such as acetic acid or trifluoroacetic acid, sulfonic acids such as methanesulfonic acid, trifluoromethanesulfonic or p-toluenesulfonic acid, Hydrochloric acid, Sulfuric acid, Phosphoric acid, phosphonic or phosphoric acid or phosphonic anhydrides like polyphosphoric acid, polyphosphate or propanephosphonic anhydride. Preference is given to polyphosphoric acid ethyl ester used. The acid is generally in an amount of 0.25 mol to 100 mol, based to 1 mole of the compound (III) used.

Of the Process step (II) + (III) + (IV) → (I-A) is general in a temperature range of + 20 ° C to + 150 ° C, preferably at + 60 ° C to + 100 ° C. The Implementation can be normal, elevated or reduced pressure (e.g., from 0.5 to 5 bar). In general you work at normal pressure.

Suitable solvents for process step (IA) + (V) → (I) are customary organic solvents which do not change under the reaction conditions. These include, for example, ethers such as diethyl ether, diisopropyl ether, methyl tert-butyl ether, 1,2-dimethoxyethane, dioxane or tetrahydrofuran, hydrocarbons such as pentane, hexane, cyclohexane, benzene, toluene or xylene, halogenated hydrocarbons such as di chloromethane, 1,2-dichloroethane, trichloromethane or chlorobenzene, or other solvents such as ethyl acetate, acetone, methyl ethyl ketone, methyl tert-butyl ketone, acetonitrile, dimethyl sulfoxide, N, N-dimethylformamide, N, N'-dimethylpropyleneurea (DMPU) or N-methylpyrrolidone (NMP). It is likewise possible to use mixtures of the solvents mentioned. Preference is given to using tetrahydrofuran or dimethylformamide.

When Base for the process step (I-A) + (V) → (I) are customary inorganic or organic bases. These include preferably alkali or Alkaline earth carbonates such as lithium, sodium, potassium, calcium or cesium carbonate, Alkali alcoholates such as sodium or potassium tert-butoxide, alkali metal hydrides such as sodium or potassium hydride, amides such as lithium or potassium bis (trimethylsilyl) amide or lithium diisopropylamide, or organic amines such as triethylamine, N-methylmorpholine, N-methylpiperidine, N, N-diisopropylethylamine, Pyridine or 4-N, N-dimethylaminopyridine. Preference is given to potassium carbonate, cesium carbonate or sodium hydride. The base is generally in one Amount of from 0.1 mol to 10 mol, preferably from 1 mol to 3 mol, based on 1 mol of the compound (I-A) used.

Of the Process step (I-A) + (V) → (I) is generally preferred in a temperature range of 0 ° C to + 150 ° C at + 20 ° C up to + 80 ° C carried out. The reaction may be at normal, elevated or reduced pressure take place (for example from 0.5 to 5 bar). In general, one works at Normal pressure.

The Compounds of formulas (II), (III), (IV) and (V) are commercial available, known from the literature or can be prepared in analogy to literature methods.

The compounds of the formula (I) which can be used according to the invention can, if appropriate, also be prepared by conversions of functional groups of individual substituents, in particular those listed under R ¹ and R ² , starting from other compounds of the formula (I) obtained by the above process , These conversions are carried out by conventional methods and include, for example, reactions such as esterification, ester cleavage or hydrolysis, reduction, catalytic hydrogenation, oxidation, hydroxylation, amination or alkylation, and the introduction and removal of temporary protecting groups.

Schema 2

The method described above can be illustrated by the following reaction schemes: Scheme 1

Scheme 2

The compounds of the invention can be used for the treatment and / or prophylaxis of pulmonary arterial hypertension, for example, in left atrial or left ventricular diseases and in left-sided heart valve diseases. In addition, the compounds of the invention are for the treatment and / or prophylaxis of pulmonary arterial hypertension in chronic obstructive pulmonary disease, interstitial lung disease, sleep apnea syndrome, diseases with alveolar hypo ventilation, altitude sickness and pulmonary developmental disorders.

Farther the compounds of the invention are suitable for the treatment and / or prophylaxis of pulmonary arterial hypertension due to chronic thrombotic and / or embolic diseases, such as For example, thromboembolism of the proximal pulmonary arteries, obstruction of the distal pulmonary arteries and pulmonary embolism. Furthermore, the Compounds of the invention for the treatment and / or prophylaxis of pulmonary arterial hypertension in association with sarcoidosis, histiocytosis X or lymphangioleiomyomatosis and one by vascular compression from the outside (Lymph nodes, tumor, fibrosing mediastinitis) conditional pulmonary arterial hypertension can be used.

• • Kinase-Inhibitoren, insbesondere Tyrosinkinase-Inhibitoren, wie beispielhaft und vorzugsweise Sorafenib, Imatinib, Gefitinib oder Erlotinib;
• • NO-unabhängige, jedoch Häm-abhängige Stimulatoren der löslichen Guanylatcyclase, wie insbesondere die in WO 00/06568 , WO 00/06569 , WO 02/42301 und WO 03/095451 beschriebenen Verbindungen;
• • NO- und Häm-unabhängige Aktivatoren der löslichen Guanylatcyclase, wie insbesondere die in WO 01/19355 , WO 01/19776 , WO 01/19778 , WO 01/19780 , WO 02/070462 und WO 02/070510 beschriebenen Verbindungen;
• • Prostacyclin-Analoga, wie beispielhaft und vorzugsweise Iloprost, Beraprost, Treprostinil oder Epoprostenol;
• • Endothelinrezeptor-Antagonisten, wie beispielhaft und vorzugsweise Bosentan, Darusentan, Ambrisentan oder Sitaxsentan;
• • Verbindungen, die den Abbau von cyclischem Guanosinmonophosphat (cGMP) und/oder cyclischem Adenosinmonophosphat (cAMP) inhibieren, wie beispielsweise Inhibitoren der Phosphodiesterasen (PDE) 1, 2, 3, 4 und/oder 5, insbesondere PDE 5-Inhibitoren wie Sildenafil, Vardenafil und Tadalafil.

The compounds according to the invention can be used alone or in combination with other active substances. Another object of the present invention are pharmaceutical compositions containing at least one of the compounds of the invention and one or more other active ingredients for the treatment and / or prophylaxis of the aforementioned diseases. As suitable combination active ingredients may be mentioned by way of example and preferably:

• Kinase inhibitors, in particular tyrosine kinase inhibitors, by way of example and preferably sorafenib, imatinib, gefitinib or erlotinib;
• NO-independent, but heme-dependent stimulators of soluble guanylate cyclase, such as those in WO 00/06568 . WO 00/06569 . WO 02/42301 and WO 03/095451 described compounds;
• • NO- and heme-independent activators of soluble guanylate cyclase, especially those in WO 01/19355 . WO 01/19776 . WO 01/19778 . WO 01/19780 . WO 02/070462 and WO 02/070510 described compounds;
• Prostacyclin analogs, such as by way of example and preferably iloprost, beraprost, treprostinil or epoprostenol;
• Endothelin receptor antagonists, such as by way of example and preferably bosentan, darusentan, ambrisentan or sitaxsentan;
• Compounds which inhibit the degradation of cyclic guanosine monophosphate (cGMP) and / or cyclic adenosine monophosphate (cAMP), such as inhibitors of phosphodiesterases (PDE) 1, 2, 3, 4 and / or 5, in particular PDE 5 inhibitors such as sildenafil, Vardenafil and Tadalafil.

Another The present invention is the use of the compounds of the invention alone or in combination with one or more of the aforementioned Active ingredients for the manufacture of a medicament for the treatment and / or Prophylaxis of pulmonary arterial hypertension in left atrial or left ventricular Diseases, left-sided heart valve diseases, chronic obstructive pulmonary disease, Interstitial lung disease, sleep apnea syndrome, diseases with alveolar Hypoventilation, altitude sickness, pulmonary developmental disorders, chronic thrombotic and / or embolic diseases such as For example, thromboembolism of the proximal pulmonary arteries, obstruction of the distal pulmonary arteries and pulmonary embolism, in association with Sarcoidosis, histiocytosis X or lymphangioleiomyomatosis and a by vascular compression from the outside (Lymph nodes, tumor, fibrosing mediastinitis) conditional pulmonary arterial hypertension.

Another The subject of the present invention is a method of treatment and / or prophylaxis of pulmonary arterial hypertension in humans and animals by administering an effective amount, at least one of the compounds of the invention or a medicament containing at least one of the compounds according to the invention.

The according to the use according to the invention to be produced or used according to the invention contain at least one of the compounds of the invention, usually together with one or more inert, non-toxic, pharmaceutical suitable excipients.

Another The present invention relates to medicaments containing at least one of the compounds of the invention in combination with one or more inert, non-toxic, pharmaceutical suitable excipients, for the treatment and / or prophylaxis of aforementioned diseases.

The Compounds of the invention can act systemically and / or locally. For this purpose they can be applied in a suitable manner, e.g. oral, parenteral, pulmonary, nasal, sublingual, lingual, buccal, rectal, dermal, transdermal, conjunctival, otic or as an implant or stent.

For this Application routes can the compounds of the invention be administered in suitable administration forms.

For the oral Applicable to the state of the art working, the compounds of the invention rapidly and / or modified donating application forms that the Compounds of the invention in crystalline and / or amorphised and / or dissolved form included, e.g. Tablets (non-coated or coated Tablets, for example, with enteric or delayed dissolving or insoluble coatings that control the release of the compound of the invention), in the oral cavity rapidly disintegrating tablets or films / wafers, films / lyophilisates, Capsules (for example hard or soft gelatine capsules), dragees, Granules, pellets, powders, emulsions, suspensions, aerosols or Solutions.

The Parenteral administration can bypass a resorption step happen (e.g., intravenously, intraarterial, intracardiac, intraspinal or intralumbar) or involving absorption (e.g., intramuscular, subcutaneous, intracutaneous, percutaneous or intraperitoneal). For parenteral administration are suitable as application forms u.a. Injection and infusion preparations in the form of solutions, Suspensions, emulsions, lyophilisates or sterile powders.

For the others Application routes are suitable, e.g. Inhalation medicines (i.a. Powder inhalers, nebulizers), nasal drops, solutions or sprays, lingual, sublingual or buccal tablets to be applied, films / wafers or capsules, suppositories, ear or eye preparations, vaginal capsules, aqueous Suspensions (lotions, shake mixtures), lipophilic suspensions, ointments, creams, transdermal therapeutic Systems (e.g., plasters), milk, pastes, foams, powdered powders, implants or stents.

Prefers are the oral or parenteral administration, especially oral or intravenous Application.

The Compounds of the invention can in the cited Application forms are transferred. This can be done in a conventional manner by mixing with inert, Non-toxic, pharmaceutically suitable excipients happen. Among these adjuvants include et al excipients (for example, microcrystalline cellulose, lactose, mannitol), solvent (e.g., liquid Polyethylene glycols), emulsifiers and dispersing or wetting agents (For example, sodium dodecyl sulfate, Polyoxysorbitanoleat), binder (For example, polyvinylpyrrolidone), synthetic and natural polymers (for example, albumin), stabilizers (e.g., antioxidants such as for example ascorbic acid), Dyes (e.g., inorganic pigments such as iron oxides) and flavor and / or scent remedies.

in the In general, it has proven to be beneficial in parenteral Application amounts of about 0.001 to 1 mg / kg, preferably about 0.01 to 0.5 mg / kg body weight to achieve effective results. For oral administration is the dosage about 0.01 to 100 mg / kg, preferably about 0.01 to 20 mg / kg and most preferably 0.1 to 10 mg / kg of body weight.

Nevertheless it may be necessary, if necessary, of the quantities mentioned to deviate, depending on of body weight, Route of administration, individual behavior towards the active substance, type of Preparation and time or interval to which the application he follows. So it can in some cases be sufficient, with less than the aforementioned minimum quantity to get along while in other cases exceeded the mentioned upper limit must become. In case of application of larger quantities it may be recommended be to distribute these in several single doses throughout the day.

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

The Percentages in the following tests and examples are provided not stated otherwise, weight percentages; Parts are parts by weight. Solvent ratios, dilution ratios and concentration data of liquid / liquid solutions Unless otherwise stated, each refers to the volume.

A. Examples

Abbreviations:

• aq.

  aqueous, aqueous solution

  cat.

  catalytic

  CDI

  N, N'-carbonyldiimidazole

  DC

  TLC

  DMF

  dimethylformamide

  DMSO

  dimethyl sulfoxide

  d. Th.

  the theory (at yield)

  ee

  enantiomeric excess

  eq.

  Equivalent (s)

  IT I

  Electrospray ionization (in MS)

  H

  Hour (s)

  HPLC

  High pressure, high performance liquid chromatography

  LC-MS

  Liquid chromatography-coupled Mass spectrometry

  min

  Minute (s)

  MPLC

  medium pressure

  MS

  Mass spectrometry

  NMR

  nuclear magnetic resonance spectrometry

  RT

  room temperature

  R _t

  Retention time (at HPLC)

  THF

  tetrahydrofuran

  UV

  Ultraviolet spectrometry

  v / v

  Volume to volume ratio (one Solution)

HPLC and LC-MS methods:

Method 1 (analytical HPLC):

• device type HPLC: HP 1100 Series; UV DAD; Pillar: Phenomenex Synergi 2 μ Hydro-RP Mercury 20 mm × 4 mm; Eluent A: 1 l of water + 0.5 ml of 50% formic acid, eluent B: 1 l of acetonitrile + 0.5 ml of 50% formic acid; Gradient: 0.0 min 90% A → 2.5 min 30% A → 3.0 min 5% A → 4.5 min 5% A; Flow: 0.0 min 1 ml / min → 2.5 min / 3.0 min / 4.5 min 2 ml / min; Oven: 50 ° C; UV detection: 210 nm.

Method 2 (analytical HPLC):

• device type HPLC: HP 1050 Series; UV DAD; Pillar: Phenomenex Synergi 2 μ Max-RP Mercury 20 mm × 4 mm; Eluent A: 1 liter of water + 0.05% trifluoroacetic acid, eluent B: 1 liter of acetonitrile; Gradient: 0.0 min 90% A → 2.5 min 30% A → 3.0 min 5% A → 4.5 min 5% A; Flow: 0.0 min 1 ml / min → 2.5 min / 3.0 min / 4.5 min 2 ml / min; Oven: 50 ° C; UV detection: 210 nm.

Method 3 (analytical HPLC):

Instrument: HP 1100 with DAD detection; Column: Kromasil 100 RP-18, 60 mm × 2.1 mm, 3.5 μm; Eluent A: 5 ml HClO ₄ (70%) / L water, eluent B: acetonitrile; Gradient: 0 min 2% B → 0.5 min 2% B → 4.5 min 90% B → 6.5 min 90% B → 6.7 min 2% B → 7.5 min 2% B; Flow: 0.75 ml / min; Column temperature: 30 ° C; UV detection: 210 nm.

Method 4 (analytical HPLC):

Instrument: HP 1100 with DAD detection; Column: Kromasil 100 RP-18, 60 mm × 2.1 mm, 3.5 μm; Eluent A: 5 ml HClO ₄ (70%) / L water, eluent B: acetonitrile; Gradient: 0 min 2% B → 0.5 min 2% B → 4.5 min 90% B → 9 min 90% B → 9.2 min 2% B → 10 min 2% B; Flow: 0.75 ml / min; Column temperature: 30 ° C; UV detection: 210 nm.

Method 5 (analytical HPLC on chiral Phase):

Chiral silica gel phase based on the selector poly (N-methacryloyl-D-leucine-tert-butylamide; Column: 250 mm × 4.6 mm; Eluent: ethyl acetate; Flow: 2.0 ml / min; Temperature: 24 ° C; UV detection: 260 nm.

Method 6 (LC-MS):

Device type MS: Micromass ZQ; device type HPLC: Waters Alliance 2795 / HP 1100; Column: Phenomenex Synergi 2 μ Hydro-RP Mercury 20 mm × 4 mm; Eluent A: 1 l of water + 0.5 ml of 50% formic acid, eluent B: 1 liter of acetonitrile + 0.5 ml of 50% formic acid; Gradient: 0.0 min 90% A → 2.5 min 30% A → 3.0 min 5% A → 4.5 min 5% A; Flow: 0.0 min 1 ml / min → 2.5 min / 3.0 min / 4.5 min 2 ml / min; Oven: 50 ° C; UV detection: 210 nm.

Method 7 (LC-MS):

Device type MS: Micromass ZQ; device type HPLC: Waters Alliance 2795; Pillar: Phenomenex Synergi 2 μ Hydro-RP Mercury 20 mm × 4 mm; Eluent A: 1 l of water + 0.5 ml of 50% formic acid, eluent B: 1 liter of acetonitrile + 0.5 ml of 50% formic acid; Gradient: 0.0 min 90% A → 2.5 min 30% A → 3.0 min 5% A → 4.5 min 5% A; Flow: 0.0 min 1 ml / min → 2.5 min / 3.0 min / 4.5 min 2 ml / min; Oven: 50 ° C; UV detection: 210 nm.

Method 8 (LC-MS):

Instrument: Micromass Quattro LCZ with HPLC Agilent Series 1100; Column: Phenomenex Synergi 2 μ Hydro-RP Mercury 20 mm × 4 mm; Eluent A: 1 l of water + 0.5 ml of 50% formic acid, eluent B: 1 liter of acetonitrile + 0.5 ml of 50% formic acid; Gradient: 0.0 min 90% A → 2.5 min 30% A → 3.0 min 5% A → 4.5 min 5% A; Flow: 0.0 min 1 ml / min → 2.5 min / 3.0 min / 4.5 min 2 ml / min; Oven: 50 ° C; UV detection: 208-400 nm.

Method 9 (LC-MS):

Instrument: Micromass Platform LCZ with HPLC Agilent Series 1100; Column: Thermo HyPURITY Aquastar 3 μ 50 mm × 2.1 mm; Eluent A: 1 l of water + 0.5 ml of 50% formic acid, eluent B: 1 liter of acetonitrile + 0.5 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 → 5.5 min 10% A; Oven: 50 ° C; Flow: 0.8 ml / min; UV detection: 210 nm.

Method 10 (LC-MS):

Device type MS: Micromass ZQ; device type HPLC: HP 1100 Series; UV DAD; Pillar: Phenomenex Gemini 3 μ 30 mm × 3.0 mm; Eluent A: 1 l of water + 0.5 ml of 50% formic acid, eluent B: 1 liter of acetonitrile + 0.5 ml of 50% formic acid; Gradient: 0.0 min 90% A → 2.5 min 30% A → 3.0 min 5% A → 4.5 min 5% A; Flow: 0.0 min 1 ml / min → 2.5 min / 3.0 min / 4.5 min 2 ml / min; Oven: 50 ° C; UV detection: 210 nm.

Method 11 (preparative HPLC):

Device: Gilson Abimed HPLC; binary Pump system; Pillar: Kromasil-100A C18, 5 μm, 250 mm × 21 mm; Eluent A: water / 0.5% trifluoroacetic acid, eluent B: acetonitrile; 0-10 min 10% B, ramp 10.01-55 min 100% B; Flow: 20 ml / min; UV detection: 210 nm.

Method 12 (preparative HPLC):

Device: Gilson Abimed HPLC; binary Pump system; Pillar: Kromasil-100A C18, 5 μm, 250 mm × 20 mm; Eluent A: water / 0.05% trifluoroacetic acid, eluent B: acetonitrile; 0-1 min 10% B, ramp 1.01-30 min 95% B, 30.01-34 min 95% B, 34.01-35 min 10% B; Flow: 25 ml / min; UV detection: 210 nm.

Method 13 (preparative HPLC):

Device: Gilson Abimed HPLC; binary Pump system; Pillar: Kromasil-100A C18, 5 μm, 250 mm × 20 mm; Eluent A: water / 0.1% trifluoroacetic acid, eluent B: acetonitrile; 0-1 min 10% B, ramp 1.01-40 min 90% B, 40-45 min 90% B; Flow: 25 ml / min; UV detection: 210 nm.

Method 14 (preparative HPLC):

Device: Gilson Abimed HPLC; binary pump system; Column: GromSil 120 ODS-4HE, 250 mm × 40 mm, 10 μm; Eluent A: water, eluent B: acetonitrile; 0-3 min 10% B, ramp 3.01-27 min 98% B, 27.01-34 min 98% B, 34.01-38 min 10% B; Flow: 50 ml / min; UV detection: 214 nm.

Method 15 (preparative HPLC):

Instrument: Abimed Gilson Pump 305/306, Manometric Module 806; Column: GromSil 120 ODS-4HE, 10 μm, 250 mm × 30 mm; Eluent A: water, eluent B: acetonitrile; Gradient: 0.0 min 30% B → 3 min 30% B → 31 min 95% B → 44 min 95% B → 45 min 30% B; Flow: 50 ml / min; Column temperature: RT; UV detection: 210 nm.

Method 16 (preparative HPLC on chiral phase):

Enantiomerentrennung on chiral silica gel phase based on the selector poly (N-methacryloyl-L-leucine-tert-butylamide); Pillar: 125 mm × 20 mm; the sample is dissolved in a mixture of THF / ethyl acetate 1: 5 solved; Eluent: ethyl acetate; Flow: 20 ml / min; UV detection: 260 nm; Temperature: 24 ° C.

Method 17 (preparative HPLC on chiral phase):

Enantiomerentrennung on chiral silica gel phase based on the selector poly (N-methacryloyl-L-leucine-d-menthylamide); Pillar: 250 mm × 30 mm; Eluent: Step gradient 100% ethyl acetate → 100% methanol; Flow: 50 ml / min; Temperature: 24 ° C; UV detection: 260 nm. Analytical column: 250 mm × 4.6 mm; eluent: Ethyl acetate / methanol 10: 1; Flow: 2 ml / min.

Method 18 (preparative HPLC on chiral phase):

Enantiomerentrennung on chiral silica gel phase based on the selector poly (N-methacryloyl-L-leucine-I-menthylamide); Pillar: 680 mm × 40 mm; Eluent: Step gradient 100% ethyl acetate → 100% methanol; Flow: 50 ml / min; Temperature: 24 ° C; UV detection: 260 nm. Analytical column: 250 mm × 4.6 mm; eluent: ethyl acetate; Flow: 2 ml / min.

Method 19 (preparative HPLC on chiral phase):

Enantiomerentrennung on chiral silica gel phase based on the selector poly (N-methacryloyl-L-leucine D-menthylamide); Pillar: 250 mm × 30 mm; Eluent: Step gradient 100% ethyl acetate → 100% methanol; Flow: 30 ml / min; Temperature: 24 ° C; UV detection: 260 nm. Analytical column: 250 mm × 4.6 mm; eluent: Ethyl acetate / methanol 5: 1; Flow: 2 ml / min.

Method 20 (preparative HPLC on chiral phase):

Enantiomerentrennung on chiral silica gel phase based on the selector poly (N-methacryloyl-D-leucine-3-pentylamide; Pillar: 500 mm × 63 mm; Eluent: Step gradient 100% ethyl acetate (0-16.33 min) → 100% methanol (16.34-24.12 min) → 100% Ethyl acetate (24.13-35.0 min); Flow: 100 ml / min; Temperature: 24 ° C; UV detection: 340 nm.

Method 21 (preparative HPLC on chiral phase):

Enantiomerentrennung on chiral silica gel phase based on the selector poly (N-methacryloyl-L-leucine dicyclopropylmethylamide); Pillar: 250 mm × 20 mm; Eluent: Step gradient isohexane / ethyl acetate (40:60) (0-8 min) → 100% ethyl acetate (8.01-12 min) → isohexane / ethyl acetate (40:60) (12.01-20 min); Flow: 25 ml / min; Temperature: 24 ° C; UV detection: 280 nm. Analytical Pillar: 250 mm × 4.6 mm; Eluent: ethyl acetate / isohexane 1: 1; Flow: 2 ml / min.

Method 22 (preparative HPLC):

Device: Gilson Abimed HPLC; binary Pump system; Pillar: GromSil 120 ODS-4HE, 250 mm × 40 mm, 10 μm; eluent A: water + 0.1% trifluoroacetic acid, Eluent B: acetonitrile; 10% B, ramp to 90% B in 45 min.

Starting compounds and intermediates:

Example 1A N- [3-fluoro-5- (trifluoromethyl) phenyl] urea

34 g (189,819 mmol) of 5-fluoro-3- (trifluoromethyl) aniline are dissolved in 227 ml of 2-propanol and treated at 50 ° C dropwise over a period of 10 minutes with 32.803 g (284.728 mmol) of trimethylsilyl isocyanate. The mixture is stirred overnight at 50 ° C and then concentrated on a rotary evaporator. The residue is stirred with dichloromethane, the solid is filtered off with suction and dried under high vacuum. 25.0 g (59% of theory) of the target compound are obtained.
LC-MS (Method 7): R _t = 1.69 min; MS (ESIpos): m / z (%) = 223.0 (100) [M + H] ⁺
HPLC (Method 3): R _t = 3.88 min
¹ H-NMR (400 MHz, DMSO-d ₆ ): 6 = 6.12 (br.s, 2H), 7.12 (d, 1H), 7.53 (d, 1H), 7.61 (s, 1H), 9.12 (br. s, 1H).

Example 2A N- [4-fluoro-3- (trifluoromethyl) phenyl] urea

2500 mg (13.957 mmol) of 4-fluoro-3- (trifluoromethyl) aniline are dissolved in 15 ml of 1 N hydrochloric acid and admixed with 1132 mg (13.957 mmol) of potassium cyanate. The suspension is stirred overnight at room temperature and then diluted with ethyl acetate to give a clear biphasic solution. The organic phase is separated and the aqueous extracted with ethyl acetate. After drying the combined organic phases and stripping off the solvent on a rotary evaporator, the crude product is chromatographed on silica gel (eluent: dichloromethane / methanol 80: 1, then 10: 1). 2180 mg (70% of theory) of the target compound are obtained.
LC-MS (Method 10): R _t = 1.82 min; MS (ESIpos): m / z (%) = 223.0 (100) [M + H] ⁺ .

Example 3A 4 - {(4R) -5- (1H-Imidazol-1-ylcarbonyl) -6-methyl-2-oxo-1- [3- (trifluoromethyl) phenyl] -1,2,3,4-tetrahydropyrimidine 4-yl} benzonitrile

Under an argon blanket gas atmosphere, (4R) -4- (4-cyanophenyl) -6-methyl-2-oxo-1- [3- (trifluoromethyl) phenyl] -1,2,3,4-tetrahydropyrimidine-5-carboxylic acid (2.05 g, 5 mmol; preparation according to WO 2005/082863 , Example 10A) in dry DMF (21 ml) and treated with 1,1'-carbonyldiimidazole (2.43 g, 15 mmol). The mixture is stirred for 30 min at room temperature. It is then concentrated, taken up in ethyl acetate (about 150 ml), washed with saturated aqueous sodium bicarbonate solution (about 50 ml) and saturated aqueous sodium chloride solution (50 ml), dried over sodium sulfate, filtered and concentrated in vacuo. The title compound is obtained as crude product (2.87 g, quantitative yield, about 80% purity), which is reacted without further purification.
HPLC (Method 2): R _t = 2.3 min.

EXAMPLES

Example 1 Ethyl (4R) -4- (4-cyanophenyl) -6-methyl-2-oxo-1- [3- (trifluoromethyl) phenyl] -1,2,3,4-tetrahydropyrimidine-5-carboxylate

Racemic ethyl 4- (4-cyanophenyl) -6-methyl-2-oxo-1- [3- (trifluoromethyl) phenyl] -1,2,3,4-tetrahydropyrimidine-5-carboxylate (60 g; WO 2004/024700 , Example 1) is dissolved in ethyl acetate (360 ml) and separated by chromatography into the enantiomers (Method 20). The title compound (29.9 g, 99% of th., 99.1% ee) and the isomeric ethyl (4S) -4- (4-cyanophenyl) -6-methyl-2-oxo-1- [3- (trifluoromele thyl) phenyl] -1,2,3,4-tetrahydropyrimidine-5-carboxylate.
HPLC (Method 5): R _t = 1.55 min.

The further analytical data correspond to those reported for the racemic compound (see WO 2004/024700 , Example 1).

Example 2 4 - {(4R) -5-isobutyryl-6-methyl-2-oxo-1- [3- (trifluoromethyl) phenyl] -1,2,3,4-tetrahydropyrimidin-4-yl} benzonitrile

2000 mg (15.604 mmol) of 5-methylhexane-2,4-dione, 2046 mg (15,604 mmol) of 4-formylbenzonitrile and 3186 mg (15,604 mmol) of 1- [3- (trifluoromethyl) phenyl] urea are dissolved in 60 ml of THF and with 10 g of ethyl polyphosphate (PPE). The mixture is stirred overnight at 80 ° C and then partitioned between water and ethyl acetate. The organic phase is separated, dried over sodium sulfate, filtered and the solvent is removed on a rotary evaporator. The residue is initially purified by flash chromatography on silica gel (eluent: cyclohexane / ethyl acetate 1: 1) and then by preparative HPLC on a chiral phase (method 18), the enantiomers being separated (desired enantiomer: R _t = 4.70 min). , After further purification by means of preparative HPLC (Method 15), 530 mg (8% of theory) of the title compound are obtained.
MS (ESIpos): m / z (%) = 428 (100) [M + H] ⁺
HPLC (Method 3): R _t = 8.33 min
¹ H-NMR (400 MHz, DMSO-d ₆ ): δ = 0.80 (d, 3H), 0.95 (d, 3H), 1.86 (s, 3H), 2.95 (tt, 1H), 5.48 (d, 1H) , 7.55 (d, 1H), 7.62 (d, 2H), 7.68 (t, 1H), 7.70 (s, 1H), 7.77 (d, 1H), 7.89 (d, 2H), 8.41 (d, 1H).

Example 3 4 - {(4R) -5- (Cyclobutylcarbonyl) -6-methyl-2-oxo-1- [3- (trifluoromethyl) phenyl] -1,2,3,4-tetrahydropyrimidin-4-yl} benzonitrile

The preparation of the racemic compound is carried out as in WO 2005/082864 (Example 20). The racemate is separated into the enantiomers by preparative HPLC on a chiral phase (Method 19).
HPLC (Method 19): R _t = 2.68 min
¹ H-NMR (400 MHz, DMSO-d ₆ ): δ = 1.67 (m, 114), 1.82 (m, 2H), 1.94 (s, 3H), 1.96-2.19 (m, 3H), 3.49 (dt, 114), 5.37 (d, 114), 7.53 (d, 114), 7.61 (d, 2H), 7.68 (m, 2H), 7.78 (d, 1H), 7.89 (d, 2H), 8.42 (d, 1H ).

Example 4 Tert-butyl [(6R) -6- (4-cyanophenyl) -5- (cyclobutylcarbonyl) -4-methyl-2-oxo-3- [3- (trifluoromethyl) phenyl] -3,6-dihydropyrimidine 1 (2H) -yl] acetate

170 mg (0.387 mmol) 4 - {(4R) -5- (cyclobutylcarbonyl) -6-methyl-2-oxo-1- [3- (trifluoromethyl) phenyl] -1,2,3,4-tetrahydropyrimidine-4- yl} benzonitrile are dissolved in 3 ml of DMF and treated with 91 mg (0.464 mmol) of tert-butyl bromoacetate and 96 mg (0.696 mmol) of potassium carbonate. The mixture is stirred overnight at room temperature. Subsequently, a further 91 mg (0.464 mmol) of tert-butyl bromoacetate and 50 mg (0.362 mmol) of potassium carbonate are added and the suspension is stirred at 50 ° C. for a period of six hours. For workup, the mixture is added to 30 ml of water. The aqueous phase is extracted several times with ethyl acetate, the organic phases are combined and the solvent is stripped off on a rotary evaporator. The residue is taken up in methanol and purified by preparative HPLC (Method 14). 196 mg (92% of theory) of the target compound are obtained.
LC-MS (Method 10): R _t = 3.06 min; MS (ESIpos): m / z (%) = 554.2 (18) [M + H] ⁺ .

Example 5 [(6R) -6- (4-Cyanophenyl) -5- (ethoxycarbonyl) -4-methyl-2-oxo-3- [3- (trifluoromethyl) phenyl] -3,6-dihydropyrimidine-1 (2H) -yl] acetic acid tert-butyl ester

Under an argon blanket gas atmosphere, ethyl (4R) -4- (4-cyanophenyl) -6-methyl-2-oxo-1- [3- (trifluoromethyl) phenyl] -1,2,3,4-tetrahydropyrimidine-5 carboxylate (10.74 g, 25 mmol) and solid potassium carbonate (5.53 g, 40 mmol) in dimethylformamide (100 ml). While stirring, tert-butyl bromoacetate (7.8 g, 40 mmol) is added dropwise. The reaction mixture is stirred for 20 h at room temperature and then reacted at 60 ° C for a further 3 h. The reaction mixture is concentrated in vacuo and the residue taken up in ethyl acetate (200 ml). The organic phase is then washed with water (2 × 50 ml each) and then with saturated aqueous sodium chloride solution (50 ml), dried over sodium sulfate, filtered and concentrated. The crude product is flash-chromatographed on silica gel (eluent: gradient cyclohexane → cyclohexane / ethyl acetate 6: 4). This gives 12.5 g (91% of theory) of the title compound as a solid.
LC-MS (Method 8): R _t = 2.94 min; MS (ESIpos): m / z (%) = 488.1 (100) [M + HC ₄ H ₈ ] ⁺ ;
MS (ESIneg): m / z (%) = 542.2 (100) [MH] ^-
1 H-NMR (400 MHz, DMSO-d ₆ ): δ = 1.05 (t, 3H), 1.25 (s, 9H), 2.05 (s, 3H), 3.85 (d, 1H), 4.0 (m, 3H) , 5.55 (s, 1H), 7.60-7.90 (m, 8H).

Example 6 2,3-Dihydroxypropyl (4R) -4- (4-cyanophenyl) -6-methyl-2-oxo-1- [3- (trifluoromethyl) phenyl] -1,2,3,4-tetrahydropyrimidine-5 carboxylate

200 mg (0.453 mmol) allyl (4R) -4- (4-cyanophenyl) -6-methyl-2-oxo-1- [3- (trifluoromethyl) phe nyl] -1,2,3,4-tetrahydropyrimidine-5-carboxylate (preparation according to WO 2005/082863 , Example 6A) are dissolved in 20 ml of acetone / water (3: 1). The solution is cooled to 0 ° C and treated with a solution of 72 mg (0.453 mmol) of potassium permanganate in 10 ml of acetone / water (3: 1). After stirring for 90 minutes at room temperature, saturated sodium thiosulfate solution is added and the mixture is stirred for a further 30 minutes. The workup is carried out by aspirating the batch through a kieselguhr column, which is eluted with a little methanol. After separation of the organic phase, the aqueous phase is back-extracted twice with ethyl acetate. The combined organic phases are concentrated and the residue is purified by preparative HPLC (Method 14). 119 mg (54% of theory) of the target compound are obtained.
LC-MS (Method 8): R _t = 2.06 min; MS (ESIpos): m / z (%) = 476.1 (100) [M + H] ^+
1 H-NMR (400 MHz, DMSO-d ₆ ): δ = 2.05 (s, 3H), 3.19-3.31 (m, 2H), 3.60 (m, 1H), 3.89 (ddd, 1H), 4.05 (ddd, 1H), 4.61 (q, 1H), 4.92 (dd, 1H), 5.44 (m, 1H), 7.57 (br, s, 1H), 7.63 (m, 2H), 7.70 (m, 2H), 7.79 (i.e. , 1H), 7.87 (d, 2H), 8.41 (m, 1H).

Example 7 4 - {(4R) -5-Acetyl-1- [4-fluoro-3- (trifluoromethyl) phenyl] -6-methyl-2-oxo-1,2,3,4-tetrahydropyrimidin-4-yl} benzonitrile

465 mg (4.646 mmol) of 2,4-pentanedione, 609 mg (4.646 mmol) of 4-formylbenzonitrile and 1200 mg (4.646 mmol) of N- [4-fluoro-3- (trifluoromethyl) phenyl] urea are dissolved in 12 ml of THF and with 3000 mg of ethyl polyphosphate (PPE). The mixture is stirred under reflux for six hours and then concentrated on a rotary evaporator. The residue is first purified by flash chromatography on silica gel (eluent: cyclohexane / ethyl acetate 5: 1 → 2: 1 → 1: 1) and then by preparative HPLC (method 14). The resulting racemate is separated by preparative HPLC on a chiral phase (Method 17). This gives 303 mg (16% of theory) of the title compound.
HPLC (Method 17): R ₄ = 4.70 min
¹ H-NMR (400 MHz, DMSO-d ₆ ): δ = 2.02 (s, 3H), 2.20 (s, 3H), 5.47 (d, 1H), 7.61 (m, 4H), 7.74 (br. 1H), 7.88 (d, 2H), 8.48 (d, 1H).

Example 8 [(6R) -6- (4-Cyanophenyl) -5- (cyclobutylcarbonyl) -4-methyl-2-oxo-3- [3- (trifluoromethyl) phenyl] -3,6-dihydropyrimidine-1 (2H) yl] acetic acid

190 mg (0.343 mmol) of tert-butyl [(6R) -6- (4-cyanophenyl) -5- (cyclobutylcarbonyl) -4-methyl-2-oxo-3- [3- (trifluoromethyl) phenyl] -3, Dissolve 6-dihydropyrimidine-1 (2H) -yl] acetate in 3 ml of dichloromethane, add 3 ml of trifluoroacetic acid and stir at room temperature for two hours. The reaction mixture is concentrated and the residue is purified by preparative HPLC (Method 14). 160 mg (94% of theory) of the target compound are obtained.
LC-MS (Method 10): R _t = 2.62 min; MS (ESIpos): m / z (%) = 498.1 (100) [M + H] ^+
1 H-NMR (400 MHz, DMSO-d ₆ ): δ = 1.66 (m, 1H), 1.83 (m, 2H), 1.92 (s, 3H), 2.02 (m, 2H), 2.16 (m, 1H) , 3.53 (dt, 1H), 3.84 (d, 1H), 4.18 (d, 1H), 5.60 (s, 1H), 7.59 (d, 1H), 7.65-7.74 (m, 4H), 7.81 (d, 1H ), 7.88 (d, 2H), 12.70 (s, 1H).

Example 9 5 - {(4R) -5-Acetyl-6-methyl-2-oxo-1- [3- (trifluoromethyl) phenyl] -1,2,3,4-tetrahydropyrimidin-4-yl} -pyridine-2 carbonitrile

The representation of the title connection takes place as in WO 2004/024700 (Example 74).

Example 10 2-Hydroxyethyl (4R) -4- (4-cyanophenyl) -6-methyl-2-oxo-1- [3- (trifluoromethyl) phenyl] -1,2,3,4-tetrahydropyrimidine-5-carboxylate

The representation of the title connection takes place as in WO 2004/024700 (Example 72).

Example 11 2- (Dimethylamino) ethyl (4R) -4- (4-cyanophenyl) -6-methyl-2-oxo-1- [3- (trifluoromethyl) phenyl] -1,2,3,4-tetrahydropyrimidine-5 carboxylate trifluoroacetate

Under an argon blanket gas atmosphere, 4 - {(4R) -5- (1H-imidazol-1-ylcarbonyl) -6-methyl-2-oxo-1- [3- (trifluoromethyl) phenyl] -1,2,3, 4-tetrahydropyrimidin-4-yl) benzonitrile (Example 3A, 2.26 g, 5 mmol) in N, N-dimethylethanolamine (20 ml) and then stirred at 100 ° C. for 3 h. The reaction is concentrated in vacuo, the residue is taken up in ethyl acetate (150 ml) and then washed with water (50 ml) and saturated sodium chloride solution (50 ml). The organic phase is dried over sodium sulfate, filtered and concentrated. The residue is purified by preparative HPLC (Method 22). 2.38 g (77% of theory) of the title compound are obtained.
LC-MS (Method 10): R _t = 1.2 min; MS (ESIpos): m / z (%) = 473.5 (100) [M + H] ^+
1 H-NMR (400 MHz, DMSO-d6): δ = 2.05 (s, 3H), 2.10 (br, s, 6H), 2.40 (m, 2H), 4.10 (m, 2H), 5.40 (d, 1H ), 7.55-7.70 (m, 5H), 7.80 (d, 1H), 7.90 (d, 2H), 8.40 (d, 1H).

Example 12 4 - {(4R) -5-Acetyl-1- [3-fluoro-5- (trifluoromethyl) phenyl] -6-methyl-2-oxo-1,2,3,4-tetrahydropyrimidin-4-yl} benzonitrile

2,4-pentanedione (90.1 mg, 0.9 mmol) in THF (10 ml) is placed under an argon protective gas atmosphere. 4-Cyanobenzaldehyde (118 mg, 0.9 mmol), N- [3-fluoro-5- (trifluoromethyl) phenyl] urea (200 mg, 0.9 mmol) and polyphosphoric acid ethyl ester (551 mg) are added successively. The reaction is stirred under reflux until the TLC control indicates complete conversion (approximately 4 h). The mixture is concentrated in vacuo and the residue is directly chromatographed (Biotage medium-pressure system, silica gel column, eluent: cyclohexane / ethyl acetate 5: 1 → 3: 1 → 2: 1 → 1: 1). 188 mg (50% of theory) of the racemic title compound are obtained. The racemate is then separated into the enantiomers by chromatography (Method 16). From 65 mg of the racemate, 28 mg of the enantiomerically pure title compound (> 99.5% ee) are obtained.
HPLC (Method 16): R _t = 23.27 min
LC-MS (Method 7): R _t = 2.2 min; MS (ESIpos): m / z (%) = 418.1 (100) [M + H] ^+
1 H-NMR (400 MHz, DMSO-d ₆ ): δ = 2.00 (s, 3H), 2.20 (s, 3H), 5.44 (d, 1H), 7.55 (m, 4H), 7.75 (br, s, 1H), 7.85 (d, 2H), 8.45 (d, 1H).

Example 13 [(6R) -6- (4-Cyanophenyl) -5- (ethoxycarbonyl) -4-methyl-2-oxo-3- [3- (trifluoromethyl) phenyl] -3,6-dihydropyrimidine-1 (2H) yl] acetic acid

[(6R) -6- (4-cyanophenyl) -5- (ethoxycarbonyl) -4-methyl-2-oxo-3- [3- (trifluoromethyl) phenyl] -3,6-dihydropyrimidine-1 (2H) -yl ] tert-Butyl acetate (165 mg, 0.30 mmol) is dissolved in an argon blanket gas atmosphere in Di chloromethane (1.5 ml), treated with trifluoroacetic acid (1.5 ml) and stirred for 4 h at room temperature. The reaction mixture is concentrated under reduced pressure and the residue is purified by preparative HPLC (Method 13). There are obtained 132 mg (89% of theory) of the title compound.
HPLC (method 2): R ₄ = 3.0 min
LC-MS (Method 7): R _t = 2.3 min; MS (ESIpos): m / z (%) = 488.1 (100) [M + H] ^+
1 H-NMR (400 MHz, DMSO-d ₆ ): δ = 1.10 (t, 3H), 2.05 (s, 3H), 3.75 (d, 1H), 4.05 (q, 2H), 4.10 (d, 1H) , 5.60 (s, 1H), 7.60-7.75 (m, 5H), 7.80 (d, 1H), 7.90 (d, 2H), 12.70 (s, 1H).

Example 14 [(6R) -5 - {[2- (Carboxymethoxy) ethoxy] carbonyl} -6- (4-cyanophenyl) -4-methyl-2-oxo-3- [3- (trifluoromethyl) phenyl] -3, 6-dihydropyrimidine-1 (2H) -yl] acetic acid

The representation of the title connection takes place as in WO 2005/082864 (Example 5).

Example 15 [(6R) -6- (4-Cyanophenyl) -5-isobutyryl-4-methyl-2-oxo-3- [3- (trifluoromethyl) phenyl] -3,6-dihydropyrimidin-1 (2H) -yl ] acetic acid tert-butyl ester

Sodium hydride (1.17 g, 29.2 mmol) in THF (50 ml) is placed under an argon protective gas atmosphere. A solution of the compound from Example 2 (5.0 g, 11.7 mmol) in THF (150 ml) is added slowly and the reaction mixture is stirred under foaming for 1 h at RT. Subsequently, bromoacetic acid tert-butyl ester (3.4 g, 17.5 mmol) is added slowly. After 2 h, the thin-layer chromatogram shows complete conversion. The mixture is carefully mixed with water (300 ml). The aqueous phase is saturated with solid sodium chloride and extracted three times with ethyl acetate. The combined organic Pha are dried over sodium sulfate, filtered and concentrated in vacuo. The residue is applied to silica gel and purified by MPLC (200 g of silica gel, eluent: cyclohexane / ethyl acetate 4: 1). This gives 5 g of the title compound in 79% purity and 4.4 g in 70% purity, which can be finely purified by preparative HPLC.
LC-MS (Method 7): R _t = 2.8 min; MS (ESIpos): m / z (%) = 486 (100) [MC ₄ H ₈ + H] ⁺ ;
MS (ES Ineg): m / z (%) = 540 (100) [MH] ^- .

For further analytical data see the racemic compound reported ( WO 2005/082863 Example 34A).

Example 16 [(6R) -6- (4-Cyanophenyl) -5-isobutyryl-4-methyl-2-oxo-3- [3- (trifluoromethyl) phenyl] -3,6-dihydropyrimidin-1 (2H) -yl ]acetic acid

[(6R) -6- (4-cyanophenyl) -5-isobutyryl-4-methyl-2-oxo-3- [3- (trifluoromethyl) phenyl] -3,6-dihydropyrimidine-1 (2H) -yl] acetic acid tert-butyl ester (4.43 g, 8.2 mmol) is initially charged under argon protective gas atmosphere in dichloromethane (50 ml), treated with trifluoroacetic acid (18.7 g, 164 mmol) and stirred for about 4 h at room temperature (until the DC control complete Sales indicates). The reaction mixture is concentrated in vacuo and the residue is purified by chromatography (Biotage chromatography unit, silica gel, eluent: dichloromethane → dichloromethane / methanol 100: 1 → 50: 1). There are obtained 1.44 g (34% of theory) of the title compound.
LC-MS (Method 7): R _t = 2.3 min; MS (ESIpos): m / z (%) = 486.1 (100) [M + H] ⁺ ; MS (ES Ineg): m / z (%) = 484.2 (80) [MH] ^- , 969.5 (100)
¹ H-NMR (400 MHz, DMSO-d6): 8 = 0.60 (d, 3H), 0.70 (d, 3H), 1.60 (s, 3H), 2.70 (m, 1H), 3.60 (d, 1H), 4.00 (d, 1H), 5.50 (s, 1H), 7.35-7.50 (m, 5H), 7.60 (d, 1H), 7.65 (d, 2H), 12.50 (br, s, 1H).

Example 17 3 - {[(6R) -5-Acetyl-6- (4-cyanophenyl) -4-methyl-2-oxo-3- [3- (trifluoromethyl) phenyl] -3,6-dihydropyrimidine-1 (2H ) -yl] methyl} benzoic acid

The representation of the title connection takes place as in WO 2005/082863 (Example 21).

Example 18 4 - {[(6R) -5-Acetyl-6- (4-cyanophenyl) -4-methyl-2-oxo-3- [3- (trifluoromethyl) phenyl] -3,6-dihydropyrimidine-1 (2H ) -yl] methyl} benzoic acid

The representation of the title connection takes place as in WO 2005/082863 (Example 22).

B. Evaluation of Pharmacological Activity

The Pharmacological action of the compounds of the invention can be found in the shown below:

Abbreviations:

• AMC

  7-amido-4-methylcoumarin

  BNP

  brain natriuretic peptide

  BSA

  bovine serum albumin

  HEPES

  N- (2-hydroxyethyl) piperazine-N'-2-ethanesulfonic acid

  HNE

  human neutrophils elastase

  IC

  inhibitory Concentration

  me

  OSuc methoxysuccinyl

  NADP

  Nicotinamide adenine dinucleotide phosphate

  v / v

  Volume to volume ratio (one Solution)

  w / v

  Weight to volume ratio (one Solution)

B-1. In vitro HNE inhibition test

The potency of the compounds of the invention is determined in an in vitro inhibition test. The HNE-mediated amidolytic cleavage of a suitable peptide substrate in this case leads to a fluorescence light increase. The signal intensity of the fluorescent light is directly proportional to the enzyme activity. The effective concentration of a test compound in which half of the enzyme is inhibited (50% signal intensity of the fluorescent light) is given as an IC ₅₀ value.

Execution:

In a 384-well microtiter plate, a total of 50 μl assay buffer (0.1 M HEPES pH 7.4, 0.5 M NaCl, 0.1% w / v BSA, 1% v / v DMSO), enzyme (80 pM HNE, Serva , Heidelberg) and substrate (20 μM MeOSuc-Ala-Ala-Pro-Val-AMC, from Bachem, Weil am Rhein) in the presence and absence of the test substance for two hours at 37 ° C incubated. The fluorescent light intensity of the test mixtures is measured (ex. 380 nm, em. 460 nm). The IC ₅₀ values are determined by plotting the fluorescent light intensity versus the drug concentration.

IC ₅₀ values representative of the compounds of the invention are shown in the following table: TABLE 13 Inhibition of human neutrophil elastase (HNE) Exemplary embodiment no. IC ₅₀ [nM] 3 14.8 7 5.2 8th 4.3 9 20.0 10 4.6 13 2.1 14 0.9 16 2.9 17 3.9 18 2.9

B-2. Animal model of pulmonary arterial hypertension

Rat monocrotaline-induced pulmonary hypertension is a widely used animal model of pulmonary arterial hypertension. The pyrrolizidine alkaloid monocrotaline is metabolized to the toxic monocrotaline pyrrole after subcutaneous injection in the liver and leads to endothelial damage in the pulmonary circulation within a few days, followed by remodeling of the small pulmonary arteries (medial hypertrophy, de novo muscularization). A single subcutaneous injection is sufficient to induce severe pulmonary hypertension in rats within 4 weeks [ Cowan et al., Nature Med. 6, 698-702 (2000) ].

The model uses male Sprague-Dawley rats. On day 0 the animals receive a subcutaneous injection of 60 mg / kg monocrotaline. The treatment of the animals begins at the earliest 14 days after the monocrotalin injection and extends over a period of at least 14 days. At the end of the study hemodynamic examinations of the animals as well as a determination of arterial and central venous oxygen saturation are carried out. For hemodynamic measurement, the rats are initially anesthetized with pentobarbital (60 mg / kg). Thereafter, the animals are tracheotomized and artificially ventilated (frequency: 60 breaths / min, inspiration to expiration ratio: 50:50, positive end expiratory pressure: 1 cm H ₂ O, tidal volume: 10 ml / kg body weight, FIO ₂ : 0.5). The anesthesia is maintained by isoflurane inhalation anesthesia. Systemic blood pressure is determined in the left carotid artery using a Millar microtip catheter. A polyethylene catheter is advanced over the right v. Jugular vein into the right ventricle to determine right ventricular pressure. Cardiac output is determined by thermodilution. Following hemodynamics, the heart is removed and the ratio of right to left ventricles, including the septum, is determined. Furthermore, plasma samples for the determination of biomarkers (for example, proBNP) and plasma substance levels are obtained.

B-3. CYP Inhibition Assay

The ability of substances to inhibit CYP1A2, CYP2C9, CYP2D6 and CYP3A4 in humans is investigated using pooled human liver microsomes as the enzyme source in the presence of standard substrates (see below) that form CYP-specific metabolites. Inhibition effects are assayed at six different concentrations of the test compounds (2.8, 5.6, 8.3, 16.7, 25 and 50 μM), compared with the extent of CYP-specific metabolite formation of the standard substrates in the absence of the test compounds and the corresponding IC ₅₀ values calculated. A standard inhibitor that specifically inhibits a single CYP isoform is always incubated to make results comparable between different series.

Execution:

Incubation of phenacetin, diclofenac, tolbutamide, dextromethorphan or midazolam with human liver microsomes in the presence of six different concentrations of each test compound (as a potential inhibitor) is performed on a workstation (Tecan, Genesis, Crailsheim, Germany). Standard incubation mixtures contain 1.3 mM NADP, 3.3 mM MgCl ₂ × 6 H ₂ O, 3.3 mM glucose-6-phosphate, glucose-6-phosphate dehydrogenase (0.4 U / ml) and 100 mM phosphate buffer (pH 7.4) in a total volume of 200 μl. Test compounds are preferably dissolved in acetonitrile. 96-well plates are incubated for a defined time at 37 ° C with pooled human liver microsomes. The reactions are stopped by adding 100 μl of acetonitrile, which is a suitable internal standard. Precipitated proteins are separated by centrifugation, the supernatants are pooled and analyzed by LC-MS / MS.

B-4. Hepatocyte assay for determination of metabolic stability

The metabolic stability of test compounds to hepatocytes is determined by incubating the compounds at low concentrations (preferably below 1 μM) and at low cell counts (preferably at 1 x 10 ⁶ cells / ml) to assure linear kinetic conditions in the assay. Seven samples from the incubation solution are taken at a fixed time interval for LC-MS analysis to determine the half life (ie degradation) of the compound. From this half-life different "clearance" parameters (CL) and "F _max " values are calculated (see below).

The CL and F _max values are a measure of the phase I and phase 2 metabolism of the compound in the hepatocytes. In order to minimize the influence of the organic solvent on the enzymes in the incubation approaches, its concentration generally becomes 1% (acetonitrile) or 0.1% (DMSO) limited.

For all species and breeds, a hepatocyte cell count in the liver of 1.1 × 10 ⁸ cells / g liver is expected. CL parameters based on half-lives beyond the incubation period (typically 90 minutes) can only be considered as rough guidelines.

• (QH = speziesspezifischer Leberblutfluss).

The calculated parameters and their meaning are: F _max well-stirred [%] maximum possible bioavailability after oral administration Calculation: (1-CL _blood well-stirred / QH) x 100 CL _blood well-stirred [L / (h · kg)] calculated blood clearance (well-stirred model) Calculation: (QH · CL ' _intrinsic ) / (QH + CL' _intrinsic ) CL ' _intrinsic [ml / (min · kg)] maximum ability of the liver (the hepatocytes) to metabolize a compound (assuming that the liver blood flow is not rate-limiting) Calculation: CL ' _{intrinsic, apparent} · species-specific hepatocyte count [1.1 · 10 ⁸ / g liver] · species-specific liver weight [g / kg] CL ' _{intrinsic, apparent} [ml / (min * mg)] normalizes the elimination constant by dividing it by the hepatocyte cell number x (x x 10 ⁶ / ml) used Calculation: k _el [l / min] / (cell count [x x 10 ⁶ ] / incubation volume [ml])

• (QH = species-specific liver blood flow).

C. Exemplary embodiments for pharmaceutical compositions

The Compounds of the invention can as follows be converted into pharmaceutical preparations:

Tablet:

Composition:

• 100 mg of the compound according to the invention, 50 mg lactose (monohydrate), 50 mg corn starch (native), 10 mg polyvinylpyrrolidone (PVP 25) (BASF, Ludwigshafen, Germany) and 2 mg of magnesium stearate.
• Tablet weight 212 mg. Diameter 8 mm, radius of curvature 12 mm.

production:

The Mixture of compound according to the invention, Lactose and starch is with a 5% solution (m / m) of the PVP in water granulated. The granules will dry after drying mixed with the magnesium stearate for 5 minutes. This mixture will with a usual Pressed tablet press (format of the tablet see above). When Guideline for the compression is used a pressing force of 15 kN.

Orally administrable suspension:

Composition:

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

one Single dose of 100 mg of the compound of the invention correspond 10 ml oral suspension.

production:

The Rhodigel is suspended in ethanol, the compound of the invention is added to the suspension. While stirring the addition of the water takes place. Until the completion of the swelling of the rhododendron is stirred for about 6 h.

Orally administrable solution:

Composition:

500 mg of the compound of the invention, 2.5 g polysorbate and 97 g polyethylene glycol 400. A single dose of 100 mg of the compound of the invention correspond to 20 g of oral solution.

production:

The inventive compound is suspended in the mixture of polyethylene glycol and polysorbate with stirring. The stirring process will be up to the full resolution the compound of the invention continued.

iv solution:

The inventive compound becomes in a concentration below the saturation solubility in a physiological acceptable solvent (e.g., isotonic saline, glucose solution 5% and / or PEG 400 solution 30%) solved. The solution is sterile filtered and placed in sterile and pyrogen-free injection containers bottled.

Claims (9)

Use of a compound of the formula (I)

in which X is CH or N, R ^{1 is} hydrogen, a group of the formula - (CH ₂ ) _n C (= O) -OR ^1A or a group of the formula

where * is the point of attachment to the N atom, n is the number 1 or 2 and R ^{1A is} hydrogen or (C ₁ -C ₄ ) -alkyl, R ^{2 is} cyano or a group of the formula -C (= O ) -R ^2A or -C (= O) -OR ^2A , in which R ^{2A is} (C ₁ -C ₆ ) -alkyl or (C ₃ -C ₆ ) -cycloalkyl, which in turn is up to twice, identically or differently, with hydroxy, (C ₁ -C ₄ ) -alkoxy, hydroxycarbonyl, amino, Mono- and / or di- (C ₁ -C ₄ ) -alkylamino and in which in each case a CH ₂ group may be replaced by an O atom, and R ^{3 is} either hydrogen and R ^{4 is} hydrogen, fluorine or Chloro or R ^{3 is} fluorine or chlorine and R ^{4 is} hydrogen, or one of its salts, solvates or solvates of the salts for the manufacture of a medicament for the treatment and / or prophylaxis of pulmonary arterial hypertension. Use according to claim 1 of a compound of the formula (I) in which X is CH or N, R ^{1 is} hydrogen, a group of the formula -CH ₂ -C (= O) -OH or a group of the formula

in which * is the point of attachment to the N atom, R ^{2 is} cyano, acetyl, cyclobutylcarbonyl, methoxycarbonyl, ethoxycarbonyl or 2-hydroxyethoxycarbonyl, R ^{3 is} hydrogen and R ^{4 is} hydrogen or fluorine, or one of its salts, Solvates or solvates of the salts. Use according to claim 1 or 2 of a compound according to formula (I) selected from the group consisting of compounds consisting of

and their salts, solvates and solvates of the salts. Combination containing at least one compound of the formula (I) as defined in any one of claims 1 to 3, and one or more further active ingredients selected from the group consisting from kinase inhibitors, stimulators and activators of the soluble Guanylate cyclase, prostacyclin analogs, endothelin receptor antagonists and phosphodiesterase inhibitors. Use of the combination according to claim 4 for the production of a medicament for the treatment and / or prophylaxis of the pulmonary arterial hypertension. Use of a compound of formula (I) as defined in any one of claims 1 to 3 or one A combination as defined in claim 4 for the manufacture of a medicament for the treatment and / or prophylaxis of pulmonary arterial hypertension in left atrial or left ventricular diseases, left-sided heart valve diseases, chronic obstructive pulmonary disease, interstitial lung disease, sleep apnea syndrome, diseases involving alveolar hypoventilation, altitude sickness, Pulmonary developmental disorders, chronic thrombotic and / or embolic diseases, in connection with sarcoidosis, histiocytosis X or Lymphangioleiomyomatose as well as by external vascular compression pulmonary arterial hypertension. Medicament containing a combination as in claim 4 defined. Medicament according to claim 7 for the treatment and / or Prophylaxis of pulmonary arterial hypertension. Process for the treatment and / or prophylaxis of pulmonary arterial hypertension in humans and animals Administration of an effective amount of at least one compound Ver of the formula (I) as defined in any one of claims 1 to 3 A combination as defined in claim 4 or containing a drug at least one compound of the formula (I) as in one of claims 1 to 3, in combination with one or more inert, non-toxic, pharmaceutically suitable excipients.