# Patent

## Title
1-phenyl1-3,4-dihydropyrimidin-2(1H)-one derivatives and their use

## Abstract

The invention relates to novel 1-Phenyl-3,4-dihydropyrimidin-2(1H)-one derivatives having the formula




    
    
        processes for their preparation, and their use in medicaments, especially for the treatment of chronic obstructive pulmonary diseases, acute coronary syndrome, acute myocardial infarction and heart failure development.

## Background
The present invention relates to novel heterocyclic derivatives, processes for their preparation, and their use in medicaments, especially for the treatment of chronic obstructive pulmonary diseases, acute coronary syndrome, acute myocardial infarction and heart failure development.
The fibrous protein elastin, which comprises an appreciable percentage of all protein content in some tissues, such as the arteries, some ligaments, the lungs and the heart, can be hydrolysed or otherwise destroyed by a select group of enzymes classified as elastases. Human leukocyte elastase (HLE, EC 3.4.21.37), also known as human neutrophil elastase (HNE), is a glycosylated, strongly basic serine protease and is found in the azurophilic granules of human polymorphonuclear leukocytes (PMN). HNE is released from activated PMN and has been implicated causally in the pathogenesis of acute and chronic inflammatory diseases. HNE is capable of degrading a wide range of matrix proteins including elastin and collagen, and in addition to these actions on connective tissue HNE has a broad range of inflammatory actions including upregulation of IL-8 gene expression, oedema formation, mucus gland hyperplasia and mucus hypersecretion. It also acts as a mediator of tissue injury by hydrolysing collagen structures, e.g. in the heart after acute myocardial infarction or during the development of heart failure, thus damaging endothelial cells, promoting extravasation of neutrophils adhering to the endothelium and influencing the adhesion process itself.
Pulmonary diseases where HNE is believed to play a role include lung fibrosis, pneumonia, acute respiratory distress syndrome (ARDS), pulmonary emphysema, including smoking-induced emphysema, chronic obstructive pulmonary diseases (COPD) and cystic fibrosis. In cardiovascular diseases, HNE is involved in the enhanced generation of ischaemic tissue injury followed by myocardial dysfunction after acute myocardial infarction and in the remodelling processes occurring during the development of heart failure. HNE has also been causally implicated in rheumatoid arthritis, atherosclerosis, brain trauma, cancer and related conditions in which neutrophil participation is involved.
Thus, inhibitors of HLE activity can be potentially useful in the treatment of a number of inflammatory diseases, especially of chronic obstructive pulmonary diseases [R. A. Stockley, Neutrophils and protease/antiprotease imbalance, Am. J. Respir. Crit. Care 160, S49-S52 (1999)]. Inhibitors of HLE activity can also be potentially useful in the treatment of acute myocardial syndrome, unstable angina pectoris, acute myocardial infarction and coronary artery bypass grafts (CABG) [C. P. Tiefenbacher et al., Inhibition of elastase improves myocardial function after repetitive ischaemia and myocardial infarction in the rat heart, Eur. J. Physiol. 433, S563-S570 (1997); Dinerman et al., Increased neutrophil elastase release in unstable angina pectoris and acute myocardial infarction, J. Am. Coll. Cardiol. 15, 1559-1563 (1990)], of the development of heart failure [S. J. Gilbert et al., Increased expression of promatrix metalloproteinase-9 and neutrophil elastase in canine dilated cardiomyopathy, Cardiov. Res. 34, S377-S383 (1997)] and of atherosclerosis [Dollery et al., Neutrophil elastase in human atherosclerotic plaque, Circulation 107, 2829-2836 (2003)].
The synthesis of certain 6-methyl-1,4-diphenyl-3,4-dihydro-2(1H)-pyrimidinethione derivatives is described in J. Comb. Chem. 3, 624-630 (2001), J. Fluorine Chem. 90, 17-21 (1998) and Khim. Geterotsikd. Soedin. 9, 1223-1227 (1986) [Chem. Abstr. 107, 39737 (1987)]. A specific pharmacological activity of these compounds is not mentioned.
The present invention relates to compounds of the general formulas (I-A) and (I-B)




wherein

    A represents an aryl or heteroaryl ring,
    R1, R2 and R3 independently from each other represent hydrogen, halogen, nitro, cyano, C1-C6-alkyl, hydroxy or C1-C6-alkoxy, wherein C1-C6-alkyl and C1-C6-alkoxy can be further substituted with one to three identical or different radicals selected from the group consisting of halogen, hydroxy and C1-C4-alkoxy,
    R4 represents C1-C6-alkyl, C1-C6-alkylcarbonyl, C1-C6-alkoxycarbonyl, hydroxy-carbonyl, aminocarbonyl, mono- or di-C1-C4-alkylaminocarbonyl, C6-C10-arylaminocarbonyl, heteroarylcarbonyl, heterocyclylcarbonyl, heteroaryl, heterocyclyl or cyano, wherein C1-C6-alkyl, C1-C6-alkylcarbonyl, C1-C6-alkoxycarbonyl, mono- and di-C1-C4-alkylaminocarbonyl can be further sub-stituted with one to three identical or different radicals selected from the group consisting of C3-C8-cycloalkyl, hydroxy, C1-C4-alkoxy, C1-C4-alkoxy-carbonyl, hydroxycarbonyl, aminocarbonyl, mono- and di-C1-C4-alkylamino-carbonyl, C1-C4-alkylcarbonylamino, amino, mono- and di-C1-C4-alkylamino, heteroaryl, heterocyclyl, tri-(C1-C6-alkyl)-silyl and cyano,
    R5 represents C1-C4-alkyl, which can be substituted with one to three identical or different radicals selected from the group consisting of halogen, hydroxy, C1-C6-alkoxy, C2-C6-alkenoxy, C1-C6-alkylthio, amino, mono- and di-C1-C6-alkylamino, arylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and the radical —O—C1-C4-alkyl-O—C1-C4-alkyl,
    R6A represents hydrogen, C1-C6-alkylcarbonyl, C3-C8-cycloalkylcarbonyl, C1-C6-alkoxycarbon mono- or di-C1-C4-alkylaminocarbonyl, wherein C1-C6-alkylcarbonyl, C1-C6-alkoxycarbonyl, mono- and di-C1-C4-alkylamino-carbonyl can be substituted with one to three identical or different radicals selected from the group consisting of C3-C8-cycloalkyl, hydroxy, C1-C4-alkoxy, amino, mono- and di-C1-C4-alkylamino,
    R6B represents C1-C6-alkyl, which can be substituted with one to three identical or different radicals selected from the group consisting of hydroxy, C1-C4-alkoxy, amino, mono- and di-C1-C4-alkylamino, C1-C4-alkoxycarbonyl, hydroxycarbonyl, aminocarbonyl, mono- and di-C1-C4-alkylaminocarbonyl, C1-C4-alkylcarbonyloxy, aminocarbonyloxy, cyano, aryl, heteroaryl and heterocyclyl, wherein heteroaryl and heterocyclyl can be further substituted with one to two identical or different radicals selected from the group consisting of C1-C4-alkyl, hydroxy and oxo,
    R7 represents halogen, nitro, cyano, C1-C6-alkyl, hydroxy or C1-C6-alkoxy, wherein C1-C6-alkyl and C1-C6-alkoxy can be further substituted with one to three identical or different radicals selected from the group consisting of halogen, hydroxy and C1-C4-alkoxy, and
    Y1, Y2, Y3 and Y4 independently from each other represent CH or N, wherein the ring contains either 0, 1 or 2 nitrogen atoms.


The compounds according to this invention can also be present in the form of their salts, hydrates and/or solvates.
Physiologically acceptable salts are preferred in the context of the present invention.
Physiologically acceptable salts according to the invention are non-toxic salts which in general are accessible by reaction of the compounds (I) with an inorganic or organic base or acid conventionally used for this purpose. Non-limiting examples of pharmaceutically acceptable salts of compounds (1) include the alkali metal salts, e.g. lithium, potassium and sodium salts, the alkaline earth metal salts such as magnesium and calcium salts, the quaternary ammonium salts such as, for example, triethyl ammonium salts, acetates, benzene sulphonates, benzoates, dicarbonates, disulphates, ditartrates, borates, bromides, carbonates, chlorides, citrates, dihydrochlorides, fumarates, gluconates, glutamates, hexyl resorcinates, hydrobromides, hydro-chlorides, hydroxynaphthoates, iodides, isothionates, lactates, laurates, malates, maleates, mandelates, mesylates, methylbromides, methylnitrates, methylsulphates, nitrates, oleates, oxalates, palmitates, pantothenates, phosphates, diphosphates, polygalacturonates, salicylates, stearates, sulphates, succinates, tartrates, tosylates, valerates, and other salts used for medicinal purposes.
Hydrates of the compounds of the invention or their salts are stoichiometric compositions of the compounds with water, such as for example hemi-, mono-, or dihydrates.
Solvates of the compounds of the invention or their salts are stoichiometric compositions of the compounds with solvents.
The present invention includes both the individual enantiomers or diastereomers and the corresponding racemates or diastereomeric mixtures of the compounds according to the invention and their respective salts. In addition, all possible tautomeric forms of the compounds described above are included according to the present invention. The diastereomeric mixtures can be separated into the individual isomers by chromatographic processes. The racemates can be resolved into the respective enantiomers either by chromatographic processes on chiral phases or by resolution.
In the context of the present invention, the substituents, if not stated otherwise, in general have the following meaning:
Alkyl in general represents a straight-chain or branched hydrocarbon radical having 1 to 6, preferably 1 to 4 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec.-butyl, tert.-butyl, pentyl, isopentyl, hexyl, isohexyl. The same applies to radicals such as alkoxy, alkylthio, alkylamino, alkyl-carbonyl, alkoxycarbonyl and alkoxycarbonylamino.
Alkoxy illustratively and preferably represents methoxy, ethoxy, n-propoxy, iso-propoxy, tert.-butoxy, n-pentoxy and n-hexoxy.
Alkenoxy illustratively and preferably represents allyloxy, but-2-en-1-oxy, pent-3-en-1-oxy und hex-2-en-1-oxy.
Alkylthio illustratively and preferably represents methylthio, ethylthio, n-propylthio, isopropylthio, tert.-butylthio, n-pentylthio and n-hexylthio.
Alkylcarbonyl in general represents a straight-chain or branched hydrocarbon radical having 1 to 6, preferably 1 to 4 carbon atoms which is bonded via a carbonyl group. Non-limiting examples include acetyl, n-propionyl, n-butyryl, isobutyryl, pivaloyl, n-hexanoyl.
Alkylcarbonylamino in general represents a straight-chain or branched hydrocarbon lodical having 1 to 6, preferably 1 to 4 carbon atoms which has a carbonylamino (—CO—NH—) function at the position of attachment and which is bonded to the carbonyl group. Non-limiting examples include acetylamino, n-propionylamino, n-butyryl-amino, isobutyrylamino, pivaloylamino, n-hexanoylamino.
Alkylcarbonyloxy in general represents a straight-chain or branched hydrocarbon radical having 1 to 6, preferably 1 to 4 carbon atoms which has a carbonyloxy (—CO—O—) function at the position of attachment and which is bonded to the carbonyl group. Non-limiting examples include acetyloxy, n-propionyloxy, n-butyryloxy, iso-butyryloxy, pivaloyloxy, n-hexanoyloxy.
Cycloalkyl in general represents a cyclic saturated hydrocarbon radical having 3 to 8, preferably 3 to 6 carbon atoms. Non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl.
Cycloalkylcarbonyl represents a cycloalkyl radical having 3 to 8, preferably 3 to 6 ring carbon atoms which is bonded via a carbonyl group, illustratively and preferably representing cyclopropylcarbonyl, cyclobutylcarbonyl, cyclopentylcarbonyl, cyclohexyl-carbonyl and cycloheptylcarbonyl.
Alkoxycarbonyl illustratively and preferably represents methoxycarbonyl, ethoxy-carbonyl, n-propoxycarbonyl, isopropoxycarbonyl, tert.-butoxycarbonyl, n-pentoxy-carbonyl and n-hexoxycarbonyl.
Alkylamino represents an alkylamino radical having one or two (independently selected) alkyl substituents, illustratively and preferably representing methylamino, ethylamino, n-propylamino, isopropylamino, tert.-butylamino, n-pentylamino, n-hexyl-amino, N,N-dimethylamino, N,N-diethylamino, N-ethyl-N-methylamino, N-methyl-N-n-propylamino, N-isopropyl-N-n-propylamino, N-tert.-butyl-N-methylamino, N-ethyl-N-n-pentylamino and N-n-hexyl-N-methylamino.
Alkylaminocarbonyl represents an alkylaminocarbonyl radical having one or two (independently selected) alkyl substituents, illustratively and preferably representing methylaminocarbonyl, ethylaminocarbonyl, n-propylaminocarbonyl, isopropylamino-carbonyl tert.-butylaminocarbonyl, n-pentylaminocarbonyl, n-hexylaminocarbonyl, N,N-dimethylaminocarbonyl, N,N-diethylaminocarbonyl, N-ethyl-N-methylaminocarbonyl, N-methyl-N-n-propylaminocarbonyl, N-isopropyl-N-n-propylaminocarbonyl, N-tert.-butyl-N-methylaminocarbonyl, N-ethyl-N-n-pentylamino-carbonyl and N-n-hexyl-N-methylaminocarbonyl.
Aryl represents a mono- to tricyclic aromatic carbocyclic radical having generally 6 to 14 carbon atoms, illustratively and preferably representing phenyl, naphthyl and phenanthrenyl. The same applies to radicals such as arylamino.
Heteroaryl per se and in heteroarylcarbonyl represents an aromatic mono- or bicyclic radical having generally 5 to 10 and preferably 5 or 6 ring atoms and up to 5 and preferably up to 4 heteroatoms selected from the group consisting of S, O and N, illustratively and preferably representing thienyl, furyl, pyrrolyl, thiazolyl, oxazolyl, isothiazolyl, isoxazolyl, imidazolyl, pyridyl, pyrimidyl, pyridazinyl, indolyl, indazolyl, benzofuranyl, benzothiophenyl, quinolinyl, isoquinolinyl.
Heteroarylcarbonyl illustratively and preferably represents thienylcarbonyl, furyl-carbonyl, pyrrolylcarbonyl, thiazolylcarbonyl, oxazolylcarbonyl, isoxazolylcarbonyl, imidazolylcarbonyl, pyridylcarbonyl, pyrimidylcarbonyl, pyridazinylcarbonyl, indolyl-carbonyl, indazolylcarbonyl, benzofuranylcarbonyl, benzothiophenylcarbonyl, quino-linylcarbonyl, isoquinolinylcarbonyl.
Heterocyclyl per se and in heterocyclylcarbonyl represents a mono- or polycyclic, preferably mono- or bicyclic, nonaromatic heterocyclic radical having generally 4 to 10 and preferably 5 to 8 ring atoms and up to 3 and preferably up to 2 heteroatoms and/or hetero groups selected from the group consisting of N, O, S, SO and SO2. The heterocyclyl radicals can be saturated or partially unsaturated. Preference is given to 5- to 8-membered monocyclic saturated heterocyclyl radicals having up to two heteroatoms selected from the group consisting of O, N and S, such as illustratively and preferably tetrahydrofuranyl, pyrrolidinyl, piperidinyl, morpholinyl, thio-morpholinyl, piperazinyl, perhydroazepinyl.
Heterocyclylcarbonyl illustratively and preferably represents tetrahydrofiran-carbonyl, pyrrolidinecarbonyl, piperidinecarbonyl, morpholinecarbonyl, thiomorpho-linecarbonyl, piperazinecarbonyl, perhydroazepinecarbonyl.
Halogen represents fluorine, chlorine, bromine and iodine.
When stated, that Y1, Y2, Y3 and Y4 represent CH or N, CH shall also stand for a ring carbon atom, which is substituted with a substituent R3.
In another embodiment, the present invention relates to compounds of general formulas (I-A) and (I-B),
wherein


    A represents an aryl or heteroaryl ring,
    R1, R2 and R3 independently from each other represent hydrogen, halogen, nitro, cyano, C1-C6-alkyl, hydroxy or C1-C6-alkoxy, wherein C1-C6-alkyl and C1-C6-alkoxy can be further substituted with one to three identical or different radicals selected from the group consisting of halogen, hydroxy and C1-C4-alkoxy,
    R4 represents C1-C6-alkylcarbonyl, C1-C6-alkoxycarbonyl, hydroxycarbonyl, aminocarbonyl, mono- or di-C1-C4-alkylaminocarbonyl, C6-C10-arylamino-carbonyl, heteroarylcarbonyl, heterocyclylcarbonyl, heteroaryl, heterocyclyl or cyano, wherein C1-C6-alkylcarbonyl, C1-C6-alkoxycarbonyl, mono- and di-C1-C4-alkylaminocarbonyl can be further substituted with one to three identical or different radicals selected from the group consisting of C3-C8-cycloalkyl, hydroxy, C1-C4-alkoxy, C1-C4-alkoxycarbonyl, hydroxycarbonyl, aminocarbonyl, mono- and di-C1-C4-alkylaminocarbonyl, C1-C4-alkyl-carbonylamino, amino, mono- and di-C1-C4-alkylamino, heteroaryl, heterocyclyl and tri-(C1-C6-alkyl)-silyl,
    R5 represents C1-C4-alkyl, which can be substituted with one to three identical or different radicals selected from the group consisting of halogen, hydroxy, C1-C6-alkoxy, C2-C6-alkenoxy, C1-C6-alkylthio, amino, mono- and di-C1-C6-alkylamino, arylamino, hydroxycarbonyl, C1-C6-alkoxycarbonyl and the radical —O—C1-C4-alkyl-O—C1-C4-alkyl,
    R6A represents hydrogen, C1-C6-alkylcarbonyl, C3-C8-cycloalkylcarbonyl, Cl-C6-alkoxycarbonyl, mono- or di-C1-C4-alkylaminocarbonyl, wherein C1-C6-alkylcarbonyl, C1-C6-alkoxycarbonyl, mono- and di-C1-C4-alkylamino-carbonyl can be substituted with one to three identical or different radicals selected from the group consisting of C3-C8-cycloalkyl, hydroxy, C1-C4-alkoxy, amino, mono- and di-C1-C4-alkylamino,
    R6B represents C1-C6-aklyl, which can be substituted with one to three identical or different radicals selected from the group consisting of hydroxy, C1-C4-alkoxy, amino, mono- and di-C1-C4-alkylamino, aryl, heteroaryl and hetero-cyclyl,
    R7 represents halogen, nitro, cyano, C1-C6-alkyl, hydroxy or C1-C6-alkoxy, wherein C1-C6-alkyl and C1-C6-alkoxy can be further substituted with one to three identical or different radicals selected from the group consisting of halogen, hydroxy and C1-C4-alkoxy, and
    Y1, Y2, Y3 and Y4 independently from each other represent CH or N, wherein the ring contains either 0, 1 or 2 nitrogen atoms.


In another embodiment, the present invention relates to compounds of general formulas (I-A) and (I-B),
wherein


    A represents a phenyl or pyridyl ring,
    R1, R2 and R3 independently from each other represent hydrogen, fluoro, chloro, bromo, nitro, cyano, methyl, ethyl, trifluoromethyl or trifluoromethoxy,
    R4 represents C1-C6-alkylcarbonyl, C1-C6-alkoxycarbonyl, hydroxycarbonyl, aminocarbonyl, mono- or di-C1-C4-alkylaminocarbonyl or cyano, wherein C1-C6-alkylcarbonyl, C1-C6-aloxycarbonyl and mono-C1-C4-alkylamino-carbonyl can be substituted with one to three identical or different radicals selected from the group consisting of C3-C6-cycloalkyl, hydroxy, C1-C4-alkoxy, C1-C4-alkoxycarbonyl, amino, mono- or di-C1-C4-alylamino, hetero-aryl and heterocyclyl,
    R5 represents methyl or ethyl,
    R6A represents hydrogen, C1-C6-alkylcarbonyl or C3-C6-cycloalkylcarbonyl, wherein C1-C6-alkylcarbonyl can be substituted with a radical selected from the group consisting of C3-C6-cycloalkyl, hydroxy, C1-C4-alkoxy, amino, mono- and di-C1-C4-alkylamino,
    R6B represents C1-C6-alkyl, which can be substituted with a radical selected from the group consisting of hydroxy, C1-C4-alkoxy, amino, mono- and di-C1-C4-alkylamino, phenyl, heteroaryl and heterocyclyl,
    R7 represents halogen, nitro, cyano, trifluoromethyl, trifluoromethoxy, methyl or ethyl, and
    Y1, Y2, Y3 and Y4 each represent CH.


In another embodiment, the present invention relates to compounds of general formulas (I-A) and (I-B),
wherein


    A represents a phenyl or a pyridyl ring,
    R1 and R3 each represent hydrogen,
    R2 represents fluoro, chloro, bromo, nitro or cyano,
    R4 represents C1-C4-alkylcarbonyl or C1-C4-alkoxycarbonyl, wherein C1-C4-alk-oxycarbonyl can be substituted with a radical selected from the group consisting of hydroxy, C1-C4-alkoxy, C1-C4-alkoxycarbonyl, mono- and di-C1-C4-alkylamino, heteroaryl and heterocyclyl,
    R5 represents methyl,
    R6A represents hydrogen, C1-C6-alkylcarbonyl or C3-C6-cycloalkylcarbonyl,
    R6b represents C1-C4-alkyl, which can be substituted with a radical selected from the group consisting of hydroxy, C1-C4-alkoxy, amino, di-C1-C4-alkylamino, phenyl, pyridyl, imidazolyl, pyrrolidino and morpholino,
    R7 represents trifluoromethyl or nitro, and
    Y1, Y2, Y3 and Y4 each represent CH.


In another embodiment, the present invention relates to compounds according to general formulas (I-A) and (I-B), wherein A is phenyl or pyridyl.
In another embodiment, the present invention relates to compounds according to general formulas (I-A) and (I-B), wherein R1 is hydrogen.
In another embodiment, the present invention relates to compounds according to general formulas (I-A) and (I-B), wherein R2 is cyano, especially wherein A is phenyl or pyridyl and R2 is cyano located in para-position relative to the central di-hydropyrimidinthione ring.
In another embodiment, the present invention relates to compounds according to general formulas (I-A) and (I-B), wherein R3 is hydrogen.
In another embodiment, the present invention relates to compounds according to general formulas (I-A) and (I-B), wherein R4 is C1-C4-alkoxycarbonyl, which can be substituted with dimethylamino, diethylamino, N-ethylmethylamino, pyrrolidino or piperidino, or wherein R4 is C1-C4-alkylcarbonyl, especially methylcarbonyl. In another embodiment, the present invention relates to compounds according to general formulas (I-A) and (I-B), wherein R5 is methyl.
In another embodiment, the present invention relates to compounds according to general formula (I-A), wherein R6A is hydrogen.
In another embodiment, the present invention relates to compounds according to general formula (I-B), wherein R6B is methyl, (1H-imidazol-2-yl)methyl, 2-(diethyl-amino)ethyl, 2-hydroxyethyl, 3-hydroxypropyl and 2-(1-pyrrolidinyl)ethyl.
In another embodiment, the present invention relates to compounds according to general formulas (I-A) and (I-B), wherein R7 is trifluoromethyl or nitro.
In another embodiment, the present invention relates to compounds of general formula (I-C)




wherein

    Z represents CH or N, and R1, R3 and R4 have the meaning indicated above.


In another embodiment, the present invention relates to compounds of general formula (I-E)




wherein

    Z represents CH or N,
    R1, R3 and R4 have the meaning indicated above, and
    R6B represents C1-C4-alkyl, which can be substituted with a radical selected from the group consisting of hydroxy, di-C1-C4-alkylamino, phenyl, pyridyl, imidazolyl, pyrrolidino and morpholino.


The compounds of the present invention, wherein R6A in general formula (I-A) is hydrogen, can enolize into the corresponding mercaptoamidines:




The compounds of general formulas (I-A), (I-B), (I-C) and (I-E), respectively, can be synthesized by condensing compounds of general formula (II)




wherein A, R1 and R2 have the meaning indicated above, with compounds of general formula (III)





wherein R4 and R5 have the meaning indicated above, and compounds of general formula (IV)





wherein R3, R7, and Y1 to Y4 have the meaning indicated above, in the presence of an acid either in a three-component/one-step reaction or sequentially to give compounds of the general formula (I-D)





wherein

    A, R1to R5, R7, and Y1 to Y4 have the meaning indicated above,
    optionally followed by reaction of the compounds of general formula (I-D) in the presence of a base either
    [A] with compounds of the general formula (V)

R6A*—XA  (V),
    
        wherein R6* has the meaning of R6A as indicated above, but does not represent hydrogen, and XA represents a leaving group, such as halogen,
        to give compounds of the general formula (I-A) or (I-C), respectively, or
    
    
    [B] with compounds of the general formula (VI)

R6B—XB  (VI),
    
        wherein R6B has the meaning indicated above and XB represents a leaving group, such as halogen, tosylate, mesylate or sulfate,
        to give compounds of the general formula (I-B) or (I-E), respectively.
    
    


Suitable solvents for the process (II)+(III)+(IV)→(I-D) are generally customary organic solvents which do not change under the reaction conditions. These include ethers such as diethyl ether, diisopropyl ether, 1,2-dimethoxyethane, dioxan or tetrahydroflran, ethyl acetate, acetone, acetonitrile, dimethylsulfoxide, dimethyl-formamide, or alcohols such as methanol, ethanol, n-propanol, isopropanol, n-butanol or t-butanol, or hydrocarbons such as pentane, hexane, cyclohexane, benzene, toluene or xylene, or halogeno-hydrocarbons such as dichloromethane, dichloroethane, trichloromethane or chlorobenzene. It is also possible to use mixtures of the above-mentioned solvents. Preferred for the process is tetrahydrofuran.
Suitable acids for the process (II)+(III)+(IV)→(I-D) are generally inorganic or organic acids. These preferably include carboxylic acids, such as, for example, acetic acid or trifluoroacetic acid, sulfonic acids, such as, for example, methanesulfonic acid or p-toluenesulfonic acid, hydrochloric acid or phosphoric acids such as poly-phosphoric acids. Preference is given to polyphosphoric acid ethyl ester or poly-phosphoric acid trimethylsilyl ester. The acid is employed in an amount from 0.25 mol to 100 mol, relative to 1 mol of the compound of the general formula (III).
The process is in general carried out in a temperature range from +20° C. to +150° C., preferably from +60° C. to +100° C.
The process is generally carried out at normal pressure. However, it is also possible to carry it out at elevated pressure or at reduced pressure (for example in a range from 0.5 to 5 bar).
Suitable solvents for the process (I-D)+(V)→(I-A)/(I-C) are generally customary organic solvents which do not change under the reaction conditions. These include ethers such as diethyl ether, diisopropyl ether, 1,2-dimethoxyethane, dioxan or tetra-hydrofuran, ethyl acetate, acetone, acetonitrile, dimethylsulfoxide, dimethylform-amide, or halogeno-hydrocarbons such as dichloromethane, dichloroethane, tri-chloromethane or chlorobenzene. It is also possible to use mixtures of the above-mentioned solvents. Preferred for the process is tetrahydrofuran.
Suitable bases for the process (I-D)+(V)→(I-A)/(I-C) are generally inorganic or organic bases. These preferably include cyclic amines, such as, for example, piperidine, pyridine or 4N,N-dimethylaminopyridine, or (C1-C4)-trialkylamines, such as, for example, triethylamine or diisopropylethylamine. Preference is given to pyridine. The base is employed in an amount from 0.1 mol to 10 mol, preferably from 1 mol to 3 mol, relative to 1 mol of the compound of general formula (I-D).
The process is in general carried out in a temperature range from −20° C. to +120° C., preferably from +0° C. to +80° C., especially at room temperature.
The process is generally carried out at normal pressure. However, it is also possible to carry it out at elevated pressure or at reduced pressure (for example in a range from 0.5 to 5 bar).
Suitable solvents for the process (I-D)+(VI)→(I-B)/(I-E) are generally customary organic solvents which do not change under the reaction conditions. These include ethers such as diethyl ether, diisopropyl ether, 1,2-dimethoxyethane, dioxan or tetra-hydrofuran, ethyl acetate, acetone, acetonitrile, dimethylsulfoxide, dimethylform-amide, or halogeno-hydrocarbons such as dichloromethane, dichloroethane, tri-chloromethane or chlorobenzene. It is also possible to use mixtures of the above-mentioned solvents. Preferred for the process is acetone.
Suitable bases for the process (I-D)+(VI)→(I-B)/(I-E) are generally inorganic or organic bases. These preferably include alkali hydroxides, such as lithium, sodium or potassium hydroxide, alkali or alkaline-earth carbonates, such as sodium, potassium, calcium or caesium carbonate, alkali alkoxides, such as sodium or potassium methoxide, sodium or potassium ethoxide, or sodium or potassium tert.-butoxide, or cyclic amines, such as piperidine, pyridine or 4-N,N-dimethylaminopyridine, or (C1-C4)-trialkylamines, such as triethylamine or diisopropylethylamine, or hydrides such as sodium hydride. Preference is given to potassium carbonate. The base is employed in an amount from 0.1 mol to 10 mol, preferably from 1 mol to 3 mol, relative to 1 mol of the compound of general formula (I-D).
To enhance the reactivity of the compounds of general formula (VI) in cases where XB is chloride or bromide, the process (I-D)+(VI)→(I-B)/(I-E) is preferably carried out in the presence of catalytic amounts of iodide sources, such as potassium iodide or tetralutylammonium iodide.
The process is in general carried out in a temperature range from 0° C. to +150° C., preferably from +0° C. to +80° C., especially at room temperature.
The process is generally carried out at normal pressure. However, it is also possible to carry it out at elevated pressure or at reduced pressure (for example in a range from 0.5 to 5 bar).
The compounds of the general formulas (II), (III), (I), (V) and (VI) are known per se, or they can be prepared by customary methods.
The above-mentioned method can be illustrated by the following scheme:




The compounds according to the invention exhibit an unforeseeable, useful pharmacological and pharmacokinetic activity spectrum. They are therefore suitable for use as medicaments for the treatment and/or prophylaxis of disorders in humans and animals.
Surprisingly, the compounds of the present invention show human neutrophil elastase (HNE) inhibitory activity and are therefore suitable for the preparation of medicaments for the treatment of diseases associated with HNE activity. They may thus provide an effective treatment of acute and chronic inflammatory processes, such as rheumatoid arthritis, atherosclerosis, and especially of acute and chronic pulmonary diseases, such as lung fibrosis, cystic fibrosis, pneumonia, acute respiratory distress syndrome (ARDS), in particular pulmonary emphysema, including smoking-induced emphysema, and chronic obstructive pulmonary diseases (COPD), chronic bronchitis and bronchiectasis. The compounds of the present invention may further provide an effective treatment for cardiovascular ischaemic diseases such as acute coronary syndrome, acute myocardial infarction, unstable and stable angina pectoris, coronary artery bypass grafts (CABG) and heart failure development, for atherosclerosis, mitral valvular disease, atrial septal defects, percutaneous transluminal coronary angioplasty O?TCA), inflammation after open heart surgery and for pulmonary hypertension. They may also prove useful for an effective treatment of rheumatoid arthritis, acute inflammatory arthritis, cancer, acute pancreatitis, ulcerative colitis, periodontal disease, Chury-Strauss syndrome, acute and chronic atopic dermatitis, psoriasis, systemic lupus erythematosus, bullous pemphigus, sepsis, alcoholic hepatitis, liver fibrosis, Behcet's disease, allergic fungal sinusitis, allergic sinusitis, Crohn's disease, Kawasaki disease, glomerulonephritis, acute pyelonephritis, colorectal diseases, chronic suppurative otitis media, chronic venous leg ulcers, inflammatory bowel disease, bacterial and viral infections, brain trauma, stroke and other conditions in which neutrophil participation is involved.
The present invention further provides medicaments containing at least one compound according to the invention, preferably together with one or more pharma-cologically safe excipient or carrier substances, and also their use for the above-mentioned purposes.
The active component can act systemically and/or locally. For this purpose, it can be applied in a suitable manner, for example orally, parenterally, pulmonally, nasally, sublingually, lingually, buccally, rectally, transdermally, conjunctivally, otically or as an implant.
For these application routes, the active component can be administered in suitable application forms.
Useful oral application forms include application forms which release the active component rapidly and/or in modified form, such as for example tablets (non-coated and coated tablets, for example with an enteric coating), capsules, sugar-coated tablets, granules, pellets, powders, emulsions, suspensions, solutions and aerosols. Parenteral application can be carried out with avoidance of an absorption step (intravenously, intraarterially, intracardially, intraspinally or intralumbarly) or with inclusion of an absorption (intramuscularly, subcutaneously, intracutaneously, percutaneously or intraperitoneally). Useful parenteral application forms include injection and infusion preparations in the form of solutions, suspensions, emulsions, lyophilisates and sterile powders.
Forms suitable for other application routes include for example inhalatory pharmaceutical forms (including powder inhalers, nebulizers), nasal drops/solutions, sprays; tablets or capsules to be administered lingually, sublingually or buccally, suppositories, ear and eye preparations, vaginal capsules, aqueous suspensions (lotions, shake mixtures), lipophilic suspensions, ointments, creams, rnilk, pastes, dusting powders or implants.
The active components can be converted into the recited application forms in a manner known per se. This is carried out using inert non-toxic, pharmaceutically suitable excipients. These include inter alia carriers (for example microcrystalline cellulose), solvents (for example liquid polyethylene glycols), emulsifiers (for example sodium dodecyl sulphate), dispersing agents (for example polyvinyl-pyrrolidone), synthetic and natural biopolymers (for example albumin), stabilizers (for example antioxidants such as ascorbic acid), colorants (for example inorganic pigments such as iron oxides) or taste and/or odor corrigents.
For human use, in the case of oral administration, it is recommendable to administer doses of from 0.001 to 50 mg/kg, preferably of 0.01 mg/kg to 20 mg/kg. In the case of parenteral administration, such as, for example, intravenously or via mucous membranes nasally, buccally or inhalationally, it is recommendable to use doses of 5 0.001 mg/kg to 0.5 mg/kg.
In spite of this, it can be necessary in certain circumstances to depart from the amounts mentioned, namely as a function of body weight, application route, individual behaviour towards the active component, manner of preparation and time or interval at which application takes place. It can for instance be sufficient in some cases to use less than the aforementioned minimum amount, while in other cases the upper limit mentioned will have to be exceeded. In the case of the application of larger amounts, it can be advisable to divide them into a plurality of individual doses spread through the day.
The percentages in the tests and examples which follows are, unless otherwise stated, by weight; parts are by weight. Solvent ratios, dilution ratios and concentrations reported for liquid/liquid solutions are each based on the volume.
A. Evaluation of Physiological Activity
The potential of the compounds of the invention to inhibit neutrophil elastase activity may be demonstrated, for example, using the following assays:
I. In vitro Enzyme Assays of Human Neutrophil Elastase (HNE)
Assay Contents
assay buffer: 0.1 M HEPES-NaOH buffer pH 7.4, 0.5 M NaCl, 0.1% (w/v) bovine serum albumin;
suitable concentration (see below) of HNE (18 U/mg lyophil., #20927.01, SERVA Electrophoresis GmbH, Heidelberg, Germany) in assay buffer;
suitable concentration (see below) of substrate in assay buffer;
suitable concentration of test compounds diluted with assay buffer from a 10 mM stock solution in DMSO.


EXAMPLE A
In vitro Inhibition of HNE Using a Fluorogenic Peptide Substrate (Continuous Read-out Signal, 384 MTP Assay Format):
In this protocol, the elastase substrate MeOSuc-Ala-Ala-Pro-Val-AMC (#324740, Calbiochem-Novabiochem Corporation, Merck KGaA, Darmstadt, Germany) is used. The test solution is prepared by mixing 10 μl of test compound dilution, 20 μl of HNE enzyme dilution (final concentration 8-0.4 μU/ml, routinely 2.1 μU/ml) and 20 μl of substrate dilution (final concentration 1 mM-1 μM, routinely 20 μM), respectively. The solution is incubated for 0-2 hrs at 37° C. (routinely one hour). The fluorescence of the liberated AMC due to the enzymatic reaction is measured at 37° C. (TECAN spectra fluor plus plate reader). The rate of increase of the fluorescence (ex. 395 mm. 460 nm) is proportional to elastase activity. IC50 values are determined by RFU-versus-[I] plots. Km and Km(app.) values are determined by Lineweaver-Burk plots and converted to Ki values by Dixon plots.
The preparation examples had IC50 values within the range of 5 nM-5 μM in this assay. Representative data are given in Table 1:










TABLE 1







Example No.
IC50 [nM]



















1
13



3
9



4
23



6
70



7
10



9
40



12
100



14
10



15
5



19
30



29
10



31
80



33
20



58
12










EXAMPLE B
In vitro Inhibition of HNE Using a Fluorogenic, Unsoluble Elastin Substrate (Discontinuous Read-out Signal, 96 MTP Assay Format):
In this protocol the elastase substrate elastin-fluorescein (#100620, ICN Biomedicals GmbH, Eschwege, Germany) is used. The test solution is prepared by mixing 3 μl of test compound dilution, 77 μl of HNE enzyme dilution (final concentration 0.22 U/ml-2.2 mU/ml, routinely 21.7 μU/ml) and 80 μl substrate suspension (final concentration 2 mg/mi). The suspension is incubated for 0-16 hrs at 37° C. (routinely four hours) under slightly shaking conditions. To stop the enzymatic reaction, 160 μl of 0.1 M acetic acid are added to the test solution (final concentration 50 mM). The polymeric elastin-fluorescein is pulled down by centrifugation (Eppendorf 5804 centrifuge, 3.000 rpm, 10 nm). The supernatant is transferred into a new MIP and the fluorescence of the liberated peptide fluorescein due to the enzymatic reaction is measured (BMG Fluostar plate reader). The rate of fluorescence (ex. 490 nm, em. 520 nm) is proportional to elastase activity. IC50 values are determined by RFU-versus-[I] plots.
II. In vitro Human Neutrophil Assays
EXAMPLE A
In vitro PMN Elastolysis Assay:
This assay is used to determine the elastolytic potential of human polymorphonuclear cells (PMNs) and assess the proportion of degradation due to neutrophil elastase [cf. Z. W. She et al., Am. J. Respir. Cell. Mol. Biol. 9, 386-392 (1993)].
Tritiated elastin, in suspension, is coated on to a 96 well plate at 10 μg per well. Test and reference [ZD-0892 (J. Med. Chem. 40, 1876-1885, 3173-3181 (1997), WO 95/21855) and α1 protease inhibitor (α1PI)] compounds are added to the wells at the appropriate concentrations. Human PMNs are separated from peripheral venous blood of healthy donors and resuspended in culture media The neutrophils are added to the coated wells at concentrations ranging between 1×106 to 1×105 cells per well. Porcine pancreatic elastase (1.3 μM) is used as a positive control for the assay, and α1PI (1.2 μM) is used as the positive inhibitor of neutrophil elastase. The cellular control is PMNs without compound at each appropriate cell density. The cells plus compounds are incubated in a humidified incubator at 37° C. for 4 hours. The plates are centrifuged to allow the harvest of cell supematant only. The supernatant is transferred in 75 μl volumes to corresponding wells of a 96 well LumaplateTM (solid scintillant containing plates). The plates are dried until no liquid is visible in the wells and read in a beta counter for 3 minutes per well.
Elastolysis of the 3H-elastin results in an increase in counts in the supernatant. An inhibition of this elastolysis shows a decrease, from the cellular control, of tritium in the supernatant. α1PI gave 83.46±3.97% (mean±s.e.m.) inhibition at 1.2 μM (n=3 different donors at 3.6×105 cells per well). IC50 values were obtained for the reference compound ZD-0892 of 45.50±7.75 nM (mean±s.e.m.) (n=2 different donors at 3.6×105 cells per well).
Given that ZD-0892 is a selective inhibitor of PMN elastase along with the data from α1PI inhibition, these results indicate that the majority of elastin degradation by PMNs is due to the release of neutrophil elastase, and not to another elastolytic enzyme such as matrix metalloproteases (MMPs). The compounds of this invention are evaluated for their inhibitory activity in this HNE-dependent model of neutrophil elastolysis.
EXAMPLE B
In vitro Inhibition of Membrane Bound Elastase:
Measurement of the inhibition of elastase bound to neutrophil membranes is performed using a human neutrophil assay. Neutrophils are stimulated with LPS at 37° C. for 35 min and then spun at 1600 rpm. Subsequently, the membrane bound elastase is fixed to the neutrophils with 3% paraformaldehyde and 0.25% glutaraldehyde for 3 min at 4° C. The neutrophils are then spun, and vehicle and the compound under evaluation are added, followed by addition of the substrate MeOSuc-Ala-Ala-Pro-Val-AMC (#324740, Calbiochem-Novabiochem Corporation, Merck KGaA, Darmstadt, Germany) at 200 μM. Following a 25 min incubation at 37° C., the reaction is terminated with PMSF (phenyhnethanesulfonyl fluoride), and the fluorescence is read at ex: 400 nm and em: 505 nm. IC50 values are determined by interpolation from plots of relative fluorescence vs. inhibitor concentration.
III. In vivo Models
EXAMPLE A
In vivo Model of Acute Lung Injury in the Rat:
Instillation of human neutrophil elastase (HNE) into rat lung causes acute lung damage. The extent of this injury can be assessed by measuring lung haemorrhage.
Rats are anaesthetised with Hypnorm/Hypnovel/water and instilled with HNE or saline delivered by microsprayer into the lungs. Test compounds are administered by intravenous injection, by oral gavage or by inhalation at set times prior to the administration of HNE. Sixty minutes after the administration of elastase animals are killed by an anaesthetic overdose (sodium pentobarbitone) and the lungs lavaged with 2 ml heparinised phosphate buffered saline (PBS). Bronchoalveolar lavage (BAL) volume is recorded and the samples kept on ice. Each BAL sample is centrifuged at 900 r.p.m. for 10 minutes at 4-10° C. The supernatant is discarded and the cell pellet resuspended in PBS and the sample spun down again. The supernatant is again discarded and the cell pellet resuspended in 1 ml 0.1% cetyltrimethyl-ammonium bromide (CTAB)/PBS to lyse the cells. Samples are frozen until blood content is assayed. Prior to the haemorrhage assay the samples are defrosted and mixed. 100 μl of each sample are placed into a separate well of a 96 well flat-bottomed plate. All samples are tested in duplicate. 100 μl 0.1% CTAB/PBS is included as a blank. The absorbance of the well contents is measured at 415 nm using a spectrophotometer. A standard curve is constructed by measuring the OD at 415 nm of different concentrations of blood in 0.1% CTAB/PBS. Blood content values are calculated by comparison to the standard curve (included in each plate) and normalised for the volume of BAL fluid retrieved.
The compounds of this imvention are evaluated intravenously, orally or by inhalation for their inhibitory activity in this model of HNE-induced haemorrhage in the rat.
EXAMPLE B
In vivo Model of Acute Myocardial Infarction in the Rat:
Elastase inhibitors are tested in a rat thread infarct model. Male Wistar rats (weighing >300 g) receive 10 mg/kg aspirin 30 min prior to surgery. They are anaesthetized by isofluran and ventilated (120-130 strokes/min,.200-250 μl stroke volume; MiniVent Type 845, Hugo Sachs Elektronik, Germany) during the whole surgery. Following a left thoracotomy at the fourth intercostal space, the pericardium is opened and the heart briefly exteriorized. A thread is turned around the left coronary artery (LAD) without occluding the artery. The thread is passed under the skin to the neck of the animal. The thorax is closed and the animal is allowed to recover for 4 days. At the fifth day, rats are anaesthetized with ether for 3 min, and the thread is tied and the LAD occluded under ECG control. Test compounds are administered before or after LAD occlusion per os, intraperitoneally or intravenously (bolus or permanent infusion). After 1 hr occlusion, the thread is reopened to allow reperfusion. Hearts are excised, and infarct sizes are determined 48 hours later by staining of the re-occluded hearts with Evans blue, followed by TTC (triphenyltetrazolium chloride) staining of 2 mm heart sections. Normoxic (not occluded tissue) areas stain blue, ischemic (occluded but surviving tissue) areas stain red and necrotic (occluded dead tissue) areas remain white. Each tissue section is scanned and infarct sizes are determined by computer planimetry.
B. EXAMPLES
Abbreviations:


    DMU N,N-dimethylfobramide
    DMSO dimethylsulfoxide
    EI electron-impact ionisation (for MS)
    ESI electro-spray ionisation (for MS)
    h hour(s)
    HPLC high pressure liquid chromatography
    LC-MS liquid chromatography coupled with mass spectroscopy
    MS mass spectroscopy
    NMR nuclear magnetic resonance
    of th. of theoretical (yield)
    RP reverse phase (for HPLC)
    Rt retention time (for HPLC)
    THF tetrahydrofuran

General Methods:



All reactions were carried out under an argon atmosphere unless otherwise noted. Solvents were used as purchased from commercial sources without further purification. ‘Silica gel’ or ‘Silica’ refers to Silica gel 60 (0.040 mm-0.063 mm) from Merck KGaA company, Germany. Compounds purified over preparative HPLC were purified over a RP18-column with acetonitrile and water as the eluent, using a 1:9 to 9:1 gradient.
LC-MS and HPLC Methods:
Method 1:
Instrument: Micromass Platform LCZ, HP1100; Column: Symmetry C18, 50 mm×2.1 mm, 3.5 μm; Eluent A: water+0.05% formic acid, Eluent B: acetonitrile+0.05% formic acid; Gradient: 0.0 min 90% A→4.0 min 10% A→6.0 min 10% A; Oven: 40° C.; Flow: 0.5 ml/min; UV-detection: 208-400 nm.
Method 2:
Instrument MS: Micromass ZQ; Instrument HPLC: Waters Alliance 2790; Column: Uptisphere HDO, 50 mm×2.0 mm, 3.0 μm; Eluent A: water+0.05% formic acid, Eluent B: acetonitrile+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; Temperature: 45° C.; Flow: 0.75 ml/min; UV-detection: 210 nm.
Method 3:
Instrument: Micromass Platform LCZ, HP1100; Column: Grom-Sil 120 ODS-4 HE, 50 mm×2.0 mm, 3 μm; Eluent A: water+0.05% formic acid, Eluent B: acetonitrile+0.05% 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; Temperature: 55° C.; Flow: 0.8 ml/min; UV-detection: 208-400 nm.
Method 4:
Instrument: Micromass Quattro LCZ with HPLC Agilent Series 1100; Column: Uptisphere HDO, 50 mm×2.0 mm, 3 μm; Eluent A: 11 water+1 ml 50% formic acid, Eluent B: 11 acetonitrile+1 ml 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; Temperature: 55° C.; Flow: 0.8 ml/min; UV-detection: 208-400 nm.
Method 5:
Instrument MS: Micromass ZQ; Instrument HPLC: Waters Alliance 2790; Column: Grom-Sil 120 ODS-4 HE 50 mm×2 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; Temperature: 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 6:
Instrument MS: Micromass ZQ; Instrument HPLC: HP 1100 Series; UV DAD; Column: Grom-Sil 120 ODS-4 HE, 50 mm×2 mm, 3.0 μm; Eluent A: water+500 μl 50% formic acid/1, Eluent B: acetonitrile+500 μl 50% formic acid/1; Gradient: 0.0 min 0% B→2.9 min 70% B→3.1 min 90% B→4.5 min 90% B; Temperature: 50° C.; Flow: 0.8 ml/min; UV-detection: 210 nm.
Method 7:
Instrument: Micromass Platform LCZ with HPLC Agilent Series 1100; Column: Grom-Sil 120 ODS-4 HE, 50 mm×2.0 mm, 3 μm; Eluent A: 11 water+1 ml formic acid (50%), Eluent B: 11 acetonitrile+1 ml formic acid (50%); 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; Temperature: 55° C.; Flow: 0.8 mi/min; UV-detection: 208-400 nm.
Method 8:
Instrument: Micromass TOF-MUX-Interface quadruple parallel injection with HPLC Waters 600; Column: Uptisphere HDO, 50 mm×2.0 mm, 3.0 μm; Eluent A: 11 water+1 ml formic acid (50%), Eluent B: 11 acetonitrile+1 ml formic acid (50%); 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→4.6 min 100% A→6.5 min 100% A; Temperature: room temperature; Flow: 0.8 ml/min; UV-detection: 210 nm.
PREPARATION EXAMPLES
Example 1
4-{5-Acetyl-6-methyl-2-thioxo-1-[3-(trifiuoromethyl)phenyl]-1,2,3,4-tetrahydro-4-pyrimidinyl}benzonitrile




3-Trifluoromethylphenyl thiourea (200 mg, 0.91 mol), 4-cyanobenzaldehyde (238.2 mg, 1.82 mmol) and 2,4-pentanedione (181.9 mg, 1.82 mmol) are dissolved in 5 ml THF. Ethyl polyphosphate (0.30 g) is added and the reaction mixture is stirred at reflux temperature overnight. After cooling to room temperature, the reaction is quenched with 10 ml of water and extracted with 10 ml ethyl acetate (2×). The combined organic layers are dried with sodium sulfate and the solvent is removed in vacuo. The product is purified via preparative HPLC (RP18-column; eluent: acetonitrile-water, gradient 10:90 to 90:10).
Yield: 53 mg (14% of th.) LC-MS (method 1): Rt=4.48 min. MS (ESIpos): m/z=416 (M+H)+1H-NMR (200 MHz, DMSO-d6): δ=10.32 (d, 1H); 7.93 (d, 2H); 7.43-7.84 (m, 6H); 5.48 (d, 1H); 2.26 (s, 3H); 1.99 (s, 3H) ppm.
Example 2
Methyl 4-(4-cyanophenyl)-6-methyl-2-thioxo-1-[3-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydro-5-pyrimidinecarboxylate




3-Trifluoromethylphenyl thiourea (200 mg, 0.91 mmol), 4-cyanobenzaldehyde (238.2 mg, 1.82 mmol) and methyl acetoacetate (211 mg, 1.82 mmol) are dissolved in 5 ml THF. Ethyl polyphosphate (0.30 g) is added and the reaction mixture is stirred at reflux temperature overnight. After cooling to room temperature, the reaction is quenched with 10 ml of water and extracted with 10 ml ethyl acetate (2×). The combined organic layers are dried with sodium sulfate and the solvent is removed in vacuo. The product is purified via preparative HPLC (RP18-column; eluent: acetonitrile-water, gradient 10:90 to 90:10).
Yield: 315 mg (80% of th.) LC-MS (method 1): Rt=4.70 min. MS (ESIpos): m/z=432 (N+H)+1H-NMR (200 MHz, DMSO-d6): δ=10.24 (d, 1H); 7.92 (d, 2H); 7.54-7.83 (m, 6H); 5.40 (d, 1H); 3.63 (s, 3H); 2.06 (s, 3H) ppm.
Example 3
Ethyl 4-(4-cyanophenyl)-6-methyl-2-thioxo-1-[3-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydro-5-pyrimidinecarboxylate




3-Trifluoromethylphenyl thiourea (3.00 g, 13.6 mmol), 4-cyanobenzaldehyde (3.57 g, 27.3 mmol) and ethyl acetoacetate (3.55 g, 27.3 mmol) are dissolved in 50 ml THF. Ethyl polyphosphate (4.50 g) is added and the reaction mixture is stirred at reflux temperature overnight. After cooling to room temperature, it is quenched with 50 ml of water and extracted with 100 ml ethyl acetate (2×). The combined organic layers are dried with sodium sulfate and the solvent is removed in vacua. The product is purified via preparative HPLC (RP18-column; eluent: acetonitrile-water, gradient 10:90 to 90:10).
Yield: 3.15 g (52% of th.) LC-MS (method 2): Rt=4.10 min. MS CESIpos): m/z=446 (M+H)+1H-NMR (200 MHz, DMSO-d6): δ=10.18 (d, 1H); 7.92 (d, 2H); 7.45-7.82 (m, 6H); 5.40 (d, 1H); 4.08 (q, 2H); 2.05 (s, 3H); 1.12 (t, 3H) ppm.
Example 4
4-(4-Cyanophenyl)-6-methyl-2-thioxo-1-[3-(trlfluoromethyl)phenyl]-1,2,3,4-tetra-hydro-5-pyrimidinecarboxamide




3-Trifluoromethylphenyl thiourea (200 mg, 0.91 mmol), 4-cyanobenzaldehyde (238.2 mg, 1.82 mmol) and 3-oxobutanamide (183 mg, 1.82 mmol) are dissolved in 5 ml THF. Ethyl polyphosphate (0.30 g) is added and the reaction mixture is stirred at reflux temperature overnight. After cooling to room temperature, it is quenched with 10 ml of water and extracted with 10 ml ethyl acetate (2×). The combined organic layers are dried with sodium sulfate and the solvent is removed in vacuo. The product is purified via preparative HPLC (RP18-column; eluent: acetonitrile-water, gradient 10:90 to 90:10).
Yield: 29 mg (8% of th.) LC-MS (method 1): Rt=4.35 min. MS (ESIpos): m/z=417 (M+H)+1H-NMR (200 MHz, DMSO-d6): δ=9.86 (d, 1H); 7.93 (d, 2H); 7.76 (d, 1H); 7.67 (t, 1H); 7.24-7.62 (m, 4H); 7.59 (d, 2H); 5.40 (d, 1H); 1.74 (s, 3H) ppm.
Example 5
4-(4-Cyanophenyl)-N,6 dimethyl-2-thioxo-1-[3-(trirluoromethyl)phenyl]-1,2,3,4-tetrahydro-5-pyrimidinecarboxamide




3-Trifluoromethylphenyl thiourea (200 mg, 0.91 mmol), 4-cyanobenzaldehyde (238.2 mg, 1.82 mmol) and N-methyl-3-oxobutanamide (299 mg, 1.82 mmol) are dissolved in 5 ml THF. Ethyl polyphosphate (0.30 g) is added and the reaction mixture is stirred at reflux temperature overnight. After cooling to room temperature, the reaction is quenched with 10 ml of water and extracted with 10 ml ethyl acetate (2×). The combined organic layers are dried with sodium sulfate and the solvent is removed in vacuo. The product is purified via preparative HPLC (RP18-column; eluent: acetonitrile-water, gradient 10:90 to 90:10).
Yield: 104 mg (27% of th.) LC-MS (method 1): Rt=4.10 min MS (ESIpos): m/z=431 (M+H)+1H-NMR (200 MHz, DMSO-d6): δ=9.89 (d, 1H); 7.93 (d, 2H); 7.54 (d, 2H); 7.38-8.12 (m, 5H); 5.36 (d, 1H); 2.59 (d, 3H); 1.66 (s, 3H) ppm.
Example 6
4-(4-Cyanophenyl)-N,N,6-trimethyl-2-thioxo-1-[3-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydro-5-pyrimidinecarboxamiide




3-Trifluoromethylphenyl thiourea (200 mg, 0.91 mmol), 4-cyanobenzaldehyde (238.2 mg, 1.82 mmol) and N,N-dimethyl-3-oxobutanamide (235 mg, 1.82 mmol) are dissolved in 5 ml THF. Ethyl polyphosphate (0.30 g) is added and the reaction mixture is stirred at reflux temperature overnight. After cooling to room temperature, it is quenched with 10 ml of water and extracted with 10 ml ethyl acetate (2×). The combined organic layers are dried with sodium sulfate and the solvent is removed in vacuo. The product is purified via preparative HPLC (RP18-column; eluent: aceto-nitrile-water, gradient 10:90 to 90:10).
Yield: 270 mg (67% of th.) LC-MS (method 1): Rt=4.20 min. MS (ESIpos): m/z=445 (M+M)+1H-NMR (200 MHz, DMSO-d6): δ=9.77 (d, 1H); 7.92 (d, 2H); 7.54 (d, 2H); 7.49-7.83 (m, 4H); 5.19 (br. s, 1H); 3.33 (s, 3H); 2.78 (s, 3H); 1.43 (s, 3H) ppm.
Example 7
Ethyl 4-(4-cyanophenyl)-6-methyl-2-(methylsulfanyl)-1-[3-(trifluoromethyl)phenyl]-1,4-dihydro-5-pyrimidinecarboxylate




Ethyl 4-(4-cyanophenyl)-6-methyl-2-thioxo-1-[3-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydro-5-pyrimidinecarboxylate (Example 3; 1000 mg, 2.24 mmol), iodomethane (350.5 mg, 2.47 mmol) and potassium carbonate (341 mg, 1.82 mmol) are dissolved in 20 ml acetone. The reaction mixture is stirred at room temperature overnight and the solvent is removed in vacuo. The product is purified by column chromatography (silica gel; eluent: cyclohexane-ethyl acetate, gradient 90:10 to 50:50).
Yield: 998 mg (97% of th.) LC-MS (method 3): Rt=4.20 min. MS (ESIpos): m/z=460 (M+H)+1H-NMR (200 MHz, DMSO-d6): δ=7.85 (d, 2H); 7.69-7.94 (m, 4H); 7.60 (d, 2H); 5.76 (s, 1H); 4.04 (q, 2H); 2.14 (s, 3H); 2.01 (s, 3H); 1.11 (t, 3H) ppm.
Example 8
Methyl 4-(4-cyanophenyl)-6-methyl-2-(methylsulfanyl)1-[3-(truoromethyl)-phenyl]-1,4-dihydro-5-pyrimidinecarboxylate




Ethyl 4-(4-cyanophenyl)-6-methyl-2-(methylsulfanyl)-1-[3-(trifluoromethyl)phenyl]-1,4-dihydro-5-pyrimidinecarboxylate (Example 7; 100 mg, 0.22 mmol) and sodium methoxide (117.6 mg, 2.18 mmol) are dissolved in 5 ml methanol and stirred at reflux temperature for 3 h. The reaction is quenched with 10 ml of water and the aqueous phase is extracted with 10 ml methylene chloride (2×). After drying with sodium sulfate, the solvent is removed in vacuo and the product is purified by column chromatography (silica gel; eluent: cyclohexane-ethyl acetate, gradient 90:10 to 50:50).
Yield: 60 mg (62% of th.) LC-MS (method 2): Rt=4.27 min. LC-MS (ESIpos): m/z=446 (M+H)+.
Example 9
Ethyl 4-(4-cyanophenyl)-6-methyl-2-(ethylsulfanyl)-1-[3-trifluoromethyl)phenyl]-1,4-dihydro-5-pyrimidinecarboxylate




Ethyl 4-(4-cyanophenyl)-6-methyl-2-thioxo-1-[3-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydro-5-pyrimidinecarboxylate (Example 3; 100 mg, 0.22 mmol), iodoethane (38.5 mg, 0.25 mmol) and potassium carbonate (34.1 mg, 0.25 mmol) are dissolved in 3 ml acetone and stirred at room temperature overnight. The solvent is removed in vacuo and the product is purified via preparative HPLC (RP18-column; eluent: acetonitrile-water, gradient 10:90 to 90:10).
Yield: 62 mg (58% of th.) LC-MS (method 2): Rt=4.67 min. MS (ESIpos): m/z=474 (M+H)+1H-NMR (200 MHz, DMSO-d6): δ=7.85 (d, 2H); 7.68-7.94 (m, 4H); 7.60 (d, 2H); 5.75 (s, 1H); 4.03 (q, 2H); 2.74 (m, 2H); 2.01 (s, 3H); 1.11 (t, 3H); 1.01 (t, 3H) ppm.
Example 10
Ethyl 4-(4-cyanophenyl)-6-methyl-2-(propylsulfanyl)-1-[3-trifluoromethyl)phenyl]-1,4dihydro-5-pyrimdinecarboxylate




Ethyl 4-(4-cyanophenyl)-6-methyl-2-thioxo-1-[3-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydro-5-pyrimidinecarboxylate (Example 3; 100 mg, 0.22 mmol), 1-iodopropane (42.0 mg, 0.25 mmol) and potassium carbonate (34.1 mg, 0.25 mmol) are dissolved in 3 ml acetone and stirred at room temperature overnight. The solvent is removed in vacuo and the product is purified via preparative HPLC (RP18-column; eluent: acetonitrile-water, gradient 10:90 to 90:10).
Yield: 45 mg (41% of th.) LC-MS (method 3): Rt=4.45 min. MS (ESIpos): m/z=488 (M+H)+.
Example 11
Ethyl 4-(4-cyanophenyl)methyl-2-(butylsulfanyl)-1-[3-(trifluoromethyl)phenyl]-1,4-dihydro-5-pyrimidinecarboxylate




Ethyl 4-(4cyanophenyl)-6-methyl-2-thioxo-1-[3-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydro-5-pyrimidinecarboxylate (Example 3; 100 mg, 0.22 mmol), 1-iodobutane (45.0 mg, 0.25 mmol) and potassium carbonate (34.1 mg, 0.25 mmol) are dissolved in 3 ml acetone and stirred at room temperature overnight. The solvent is removed in vacuo and the product is purified via preparative HPLC (RP18-column; eluent: acetonitrile-water, gradient 10:90 to 90:10).
Yield: 53 mg (47% of th.) LC-MS (method 3): Rt=4.58 min. MS (ESIpos): m/z=402 (M+H)+.
Example 12
Ethyl 4-(4-cyanophenyl)-6-methyl-2-[(4-pyridinylmethyl)sulfanyl]-1-[3-(trifluoro-methyl)phenyl]-1,4-dihydro-5-pyrimidinecarboxylate




Ethyl 4-(4-cyanophenyl)-6-methyl-2-thioxo-1-[3-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydro-5-pyrimidinecarboxylate (Example 3; 100 mg, 0.22 mmol), 4-(bromo-methyl)pyridine hydrobromide (62.5 mg, 0.25 mmol), N,N,N-tributyl-1-butan-aminium iodide (7 mg, 0.03 mmol) and potassium carbonate (65.2 mg, 0.47 mmol) are dissolved in 3 ml acetone and stirred at room temperature overnight. The solvent is removed in vacuo and the product is purified via preparative HPLC (RP18-column; eluent: acetonitrile-water, gradient 10:90 to 90:10).
Yield: 34 mg (28% of th.) LC-MS (method 3): Rt=3.92 min. MS (ESIpos): m/z=537 (M+H)+1H-NMR (200 MHz, DMSO-d6): δ=8.34 (m, 2H); 7.89 (m, 2H); 7.82 (d, 2H); 7.72 (m, 2H); 7.57 (d, 2H); 7.53 (m, 1H); 7.13 (dd, 1H); 5.78 (s,1H); 3.94-4.14 (m, 4H); 2.00 (s, 3H); 1.10 (t, 3H) ppm.
Example 13
Ethyl 4-(4-cyanophenyl)-6-methyl-2-[(3-pyridinylmethyl)sulfanyl]-1-[3-(trifluoro-methyl)phenyl]-1,4-dihydro-5-pyrimidinecarboxylate




Ethyl 4-(4-cyanophenyl)-6-methyl-2-thioxo-1-[3-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydro-5-pyrimidinecarboxylate (Example 3; 100 mg, 0.22 mmol), 3-(chloro-methyl)pyridine hydrochloride (40.5 mg, 0.25 mmol), N,N,N-tributyl-1-butan-aminium iodide (7 mg, 0.03 mmol) and potassium carbonate (65.2 mg, 0.47 mmol) are dissolved in 3 ml acetone and stirred at room temperature overnight. The solvent is removed in vacuo and the product is purified via preparative HPLC (RP18-column; eluent: acetonitrile-water, gradient 10:90 to 90:10).
Yield: 25 mg (21% of th.) LC-MS (method 2): Rt=3.93 min. MS (ESIpos): m/z=537 (M+H)+.
Example 14
Ethyl 4-(4-cyanophenyl)-2-{[2-(diethylamino)ethyl]sulfanyl}-6-methyl-1-[3-(tri-fluoromethyl)phenyl]-1,4dihydro-5-pyrimidinecarboxylate




Ethyl 4-(4-cyanophenyl)-6-methyl-2-thioxo-1-[3-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydro-5-pyrimidinecarboxylate (Example 3; 100 mg, 0.22 mmol), N-(2-bromo-ethyl)-N,N-diethylamine hydrobromide (64.5 mg, 0.25 mmol), N,N,N-tributyl-1-butanaminium iodide (7 mg, 0.03 mmol) and potassium carbonate (65.2 mg, 0.47 mmol) are dissolved in 3 ml acetone and stirred at room temperature overnight. The solvent is removed in vacuo and the product is purified via preparative HPLC (RP18-column; eluent: acetonitrile-water, gradient 10:90 to 90:10).
Yield: 15 mg (12% of th.) LC-MS (method 2): Rt=3.17 min. MS (ESIpos): m/z=545 (M+H)+.
Example 15
Ethyl 4-(4cyanophenyl)2-[(1H-imidazol-2-ylmethyl)sulfanyl]-6-methyl-1-[3-(tri-fluoromethyl)phenyl]-1,4-dihydro-5-pyrimidinecarboxylate




Ethyl 4-(4-cyanophenyl)-6-methyl-2-thioxo-1-[3-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydro-5-pyrimidinecarboxylate (Example 3; 100 mg, 0.22 mmol), 2-(bromo-methyl)-1H-imidazole hydrobromide (64.5 mg, 0.25 mmol), N,N,N-tributyl-1-butan-aminium iodide (7 mg, 0.03 mmol) and potassium carbonate (65.2 mg, 0.47 mmol) are dissolved in 3 ml acetone and stirred at room temperature overnight. The solvent is removed by distillation in vacuo and the product is purified via preparative HPLC (RP18-column; eluent: acetonitrile-water, gradient 10:90 to 90:10).
Yield: 13 mg (11% of th.) LC-MS (method 3): Rt=3.74 min. MS (ESIpos): m/z=526 (M+H)+.
Example 16
2-Cyanoethyl 4-(4-cyanophenyl)-6-methyl-2-thioxo-1-[3-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydro-5-pyrimidinecarboxylate




Under argon, trimethylsilyl polyphosphate (10.6 g) is dissolved in 75.7 ml absolute dioxane. To this solution are added 4-cyanobenzaldehyde (5.95 g, 45.4 mmol), 3-trifluoromethylphenyl thiourea (5 g, 22.7 mmol) and 2-cyanoethyl acetoacetate (6.4 g, 45.4 mmol). The reaction mixture is stirred at 80° C. for 3 hours. After evaporation of the dioxane solvent, the residue is dissolved in ethyl acetate and washed with saturated aqueous sodium bicarbonate solution, 38% sodium bisulfite solution and saturated sodium chloride solution. After drying with magnesium sulfate and evaporating off the solvent, the residue is dissolved in 15 ml methanol and purified by preparative HPLC (column: Kromasil 100 C 185 μm, 30 mm×150 mm; precolumn: Gromsil ODS 4 HE 15 μm, 10 mm×20 mm; flow rate: 66 ml/min; solvent A: acetonitrile, solvent B: water; gradient: 0 min 10% A, 3 min 10% A, 11 min 90% A, 13 min 90% A, 13.2 min 10% A, 15 min 10% A; wavelength: 220 nm; injection volume: approx. 2000 μl; number of injections: 12). The product containing fractions are combined and concentrated in vacuo.
Yield: 7.73 g (72.3% of th.) MS(ESIpos): m/z=471 (M+H)+1H-NMR (200 MHz, DMSO-d6): δ=10.3 (d, 1H); 7.9 (d, 2H); 7.4-7.8 (m, 6H); 5.4 (d, 1H); 4.25 (m, 2H); 2.9 (tr, 2H); 2.1 (s, 3H) ppm.
In analogy to the procedure of Example 16, the following compounds are prepared:
















Starting
Yield
Mass


Ex.-No.
Structure
materials
[%]
[M + H]+



















17




4-cyano- benzaldehyde; 3-chlorophenyl thiourea; ethyl acetoacetate
71
412


 


18




3-nitro- benzaldehyde; 3-trifluoro- methylphenyl thiourea; ethyl acetoacetate
74
466









Example 19
4-(4-Cyanophenyl)-6-methyl-2-thioxo-1-[3-(trifluoromethyl)phenyl]-1,2,3,4-tetra-hydropyrimidine-5-carboxylic acid




2-Cyanoethyl 4-(4-cyanophenyl)-6-methyl-2-thioxo-1-[3-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydro-5-pyrimidinecarboxylate (Example 16; 47 mg, 0.1 mmol) is dissolved in 0.5 ml dioxane. After addition of 1,8-diazabicyclo[5.4.0]undec-7-ene (15 μl, 15 mg, 0.1 mmol), the reaction mixture is stirred at room temperature for 3 hours. The reaction mixture is diluted with 1 N hydrochloric acid and extracted with ethyl acetate. After drying with magnesium sulfate and evaporating off the solvent, the residue is purified by preparative HPLC (column: Nucleosil 100-5 C 18 Nautilus 20 mm×50 mm, 5 μm; solvent A: acetonitrile, solvent B: water+0.1% formic acid; flow rate: 25 ml/min; gradient: 0 min 10% A, 2 min 10% A, 6 min 90% A, 7 min 90% A, 7.1 min 10% A, 8 min 10% A; wavelength: 220 nm; injection volume: approx. 550 μl; number of injections: 1). The product containing fractions are combined and concentrated in vacuo.
Yield: 7.73 g (72.3 % of th.) MS (EI): m/z=417 (M+H)+LC-MS (method 1): Rt=4.2 min. 1H-NMR (200 MHz, DMSO-d6): δ=12.8 (broad s, 1H); 10.15 (d, 1H); 7.9 (d, 2H); 7.4-7.8 (m, 6H); 5.4 (d, 1H); 2.1 (s, 3H) ppm.
Example 20
4-{5-(1-Hydroxyethyl)-6-methyl-2-thioxo-1-[3-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydro-4-pyrimidinyl}benzonitrile




4-{5-Acetyl-6-methyl-2-thioxo-1-[3-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydro-4-pyrimidinyl}benzonitrile (Example 1; 100 mg, 0.24 mmol) is dissolved in 3 ml methanol, and sodium borohydride (10 mg, 0.26 mmol) is added. After stirring at room temperature for 1 h, the crude mixture is purified via preparative HPLC (RP18-column; eluent: acetonitrile-water, gradient 10:90 to 90:10) to give the product as a 1.6:1-mixture of the two diastereomers.
Yield: 86 mg (86% of th.) LC-MS (method 4): Rt=4.10 min. MS (ESIpos): m/z=418 (M+H)+.
Example 21
Ethyl 4-(4-cyano-2-methylphenyl)-6-methyl-2-thioxo-1-[3-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydro-5-pyrimidinecarboxylate




4-Cyano-2-methylbenzaldehyde (200 mg, 1.38 mmol), 3-trifluoromethylphenyl thiourea (276 mg, 1.25 mmol), ethyl 3-oxobutanoate (179 mg, 1.38 mmol) and trimethylsilyl polyphosphate (225 mg) in 5 ml THF are stirred overnight at reflux temperature. The reaction mixture is cooled to room temperature, and after addition of 20 ml 0.5 M hydrochloric acid the aqueous phase is extracted with ethyl acetate (2×20 ml). The combined organic layers are washed with saturated aqueous sodium carbonate solution and dried with sodium sulfate. The solvent is removed in vacuo and the crude product is purified via preparative HPLC (RP18-column; eluent: aceto-nitrile-water, gradient 10:90 to 90:10).
Yield: 320 mg (56% of th.) LC-MS (method 5): Rt=4.32 min. MS (ESIpos): m/z=460 (+H)+1H-NMR (200 MHz, DMSO-d6): δ=10.09 (d, 1H); 7.63-7.86 (m, 7H); 5.61 (d, 1H); 3.97 (q, 2H); 2.56 (s, 3); 2.08 (s, 3H); 1.00 (t, 3H) ppm.
Example 22
4-{5-Acetyl-6-methyl-3-propionyl-2-thioxo-1-[3-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydro-4-pyrimidinyl}benzonitrile




4-{5-Acetyl-6-methyl-2-thioxo-1-[3-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydro-4-pyrimidinyl}benzonitrile (Example 1; 100 mg, 0.24 mmol) is dissolved in 3 ml THF. After addition of pyridine (21 mg, 0.26 mmol) and propanoyl chloride (22 mg, 0.24 mmol), the reaction mixture is stirred at room temperature overnight and is then quenched with 10 ml water. After extraction with ethyl acetate (2×10 ml), the organic layer is dried with sodium sulfate, and the crude product is purified via flash chromatography on silica gel using a gradient of cyclohexane/ethyl acetate.
Yield: 78 mg (69% of th.) LC-MS (method 6): Rt=4.23 min. MS (ESIpos): m/z=472 (M+H)+1H-NMR (200 MHz, DMSO-d6): δ=7.44-7.97 (m, 8H); 6.66 (s, 1H); 3.28 (m, 1H); 2.87 (m, 1H); 2.45 (s, 3H); 2.09 (s, 3H); 1.17 (t, 3H) ppm.
In analogy to the procedure of Example 22, the following compounds are prepared:

















Starting
Yield
Rt [min]
Mass


Ex.-No.
Structure
materials
[%]
(method)
[M + H]+




















23




Example 1; cyclopropyl carbonyl chloride
74
4.22 (6)
484


 


24




Example 1; cyclopentyl- acetyl chloride
74
4.73 (6)
526


 


25




Example 1; cyclobutyl carbonyl chloride
42
4.44 (6)
498


 


26




Example 3; propanoyl chloride
68
4.30 (7)
502


 


27




Example 3; cyclopropyl carbonyl chloride
70
4.30 (7)
514


 


28




Example 3; cyclopentyl acetyl chloride
61
4.60 (7)
556


 


29




Example 3; cyclobutyl carbonyl chloride
59
4.40 (7)
528









Example 30
4-{5-Acetyl-6-methyl-2-(methylsulfanyl)-1-[3-(trifluoromethyl)phenyl]-1,4-dihydro-4-pyrimidinyl}benzonitrile




4-{5-Acetyl-6-methyl-2-thioxo-1-[3-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydro-4-pyrimidinyl}benzonitrile (example 1; 100 mg, 0.24 mmol), 1-iodomethane (38.0 mg, 0.27 mmol) and potassium carbonate (34.1 mg, 0.25 mmol) are dissolved in 3 ml acetone and stirred at room temperature overnight. The solvent is removed in vacuo and the crude product is purified via preparative HPLC (RP1 8-column; eluent: aceto-nitrile-water, gradient 10:90 to 90:10).
Yield: 67 mg (65% of th.) LC-MS (method 4): Rt=4.30 min. 5 MS (ESIpos): m/z=430 (M+H)+1H-NMR (200 MHz, DMSO-d6): δ=7.55-7.96 (m, 8H); 5.85 (s, 1H); 2.19 (s, 3H); 2.16 (s, 3H); 1.98 (s, 3H) ppm.
Example 31
Ethyl 4-(4-cyano-2-methylphenyl)-6-methyl-2-(methylsulfanyl)-1-[3-(trifluoro-methyl)phenyl]-1,4-dihydro-5-pyrimidinecarboxylate




Ethyl 4-(4-cyano-2-methylphenyl)-6-methyl-2-thioxo-1-[3-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydro-5-pyrimidinecarboxylate (Example 21; 100 mg, 0.22 mmol), 1-iodomethane (34.0 mg, 0.24 mmol) and potassium carbonate (34.1 mg, 0.25 mmol) are dissolved in 3 ml acetone and stirred at room temperature overnight. The solvent is removed in vacuo and the crude product is purified via preparative HPLC (RP18-column; eluent: acetonitrile-water, gradient 10:90 to 90:10).
Yield: 78 mg (76% of th.) LC-MS (method 3): Rt=4.70 min. MS (ESIpos): m/z=474 (M+H)+.
Example 32
Acetic acid 2-[5-acetyl(4-cyanophenyl)-6-methyl-1-[3-(trifluoromethyl)phenyl]-1,4-dihydropyrimidine-2-ylsulfanyl]-ethyl ester




To a solution of 2-bromoethyl acetate (37.6 mg, 0.23 mmol) in 500 μl DMF are added potassium carbonate (82.9 mg, 0.6 mmol) and 4-{5-acetyl-6-methyl-2-thioxo-1-[3-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydro-4-pyrimidinyl}benzonitrile (Example 1; 62.3 mg, 0.15 mmol). The reaction mixture is shaken for 15 hours, filtered and purified by preparative HPLC (column: Nucleosil 100-5 C 18 Nautilus 20 mm×50 mm, 5 μm; solvent A: acetonitrile, solvent B: water+0.1% formic acid; flow rate: 25 ml/min; gradient: 0 min 10% A, 2 min 10% A, 6 min 90% A, 7 min 90% A, 7.1 min 10% A, 8 nm 10% A; wavelength: 220 um; injection volume: approx. 550 μl; number of injections: 1). The product containing fractions are combined and concentrated in vacuo.
Yield: 0.6 mg (0.8% of th.) LC-MS (method 3): Rt=4.38 min. MS (ESIpos): m/z=502 (M+H)+.
In analogy to the procedure of Example 32, the following compounds are prepared:

















Starting
Yield
Rt [min]
Mass


Ex.-No.
Structure
materials
[%]
(method)
[M + H]+




















33




Example 17; carbamic acid 3-bromo- propyl ester
20.3
3.95 (7)
513


 


34




Example 3; 1-(2-chloro- ethyl)- pyrrolidine hydrochloride
3.3
3.19 (7)
543


 


35




Example 17; 1-(2-chloro- ethyl)- pyrrolidine hydrochloride
3.8
3.16 (7)
509


 


36




Example 17; 4-(2-chloro- ethyl)- morpholine hydrochloride
3.8
3.14 (7)
525


 


37




Example 17; 3-bromo- propan-1-ol
34.5
4.02 (8)
470


 


38




Example 3; 2-bromo- ethanol
25.9
 4.3 (7)
474


 


39




Example 3; 3-bromo- propan-1-ol
18.5
4.29 (3)
504


 


40




Example 1; 2-bromo- N,N-diethyl- acetamide
37.8
4.24 (3)
529


 


41




Example 17; 2-chloro- methyl-1- methyl-1H- imidazole hydrochloride
54.1
3.59 (3)
506


 


42




Example 3; 2-bromoethyl acetate
16.3
4.49 (3)
532


 


43




Example 3; bromoacetic acid
26.5
3.94 (7)
504


 


44




Example 17; iodomethane
37.6
4.44 (3)
426


 


45




Example 3; bromo- acetonitrile
49.5
3.71 (3)
485


 


46




Example 3; 2-chloro- methyl-1- methyl-1H- imidazole hydrochloride
40.8
3.13 (7)
540


 


47




Example 3; carbamic acid 3-bromo- propyl ester
14.6
3.94 (7)
547


 


48




Example 17; 2-bromo- ethanol
33.6
4.21 (3)
456


 


49




Example 3; 6-chloro- methyl-1H- pyrimidine- 2,4-dione
53.8
3.75 (7)
570


 


50




Example 1; iodoethane
24.0
4.05 (7)
444


 


51




Example 3; 2-bromo- propionitrile
13.3
3.75 (3)
499


 


52




Example 3; 2-bromo- N,N-diethyl- acetamide
51.3
4.45 (3)
559


 


53




Example 3; 2-bromo- N-methyl- acetamide
59.4
 3.9 (7)
517


 


54




Example 3; 4-chloro- methyl-3,5- dimethyl- isoxazole
60.1
4.22 (7)
555


 


55




Example 1; 4-chloro- methyl-2- methyl- thiazole hydrochloride
5.4
3.96 (7)
527


 


56




Example 1; 4-chloro- methyl-3,5- dimethyl- isoxazole
21.6
4.02 (7)
525









Example 57
4-(4-Cyanophenyl)-6-methyl-2-(methylsulfanyl)-1-[3-(trifluoromethyl)phenyl]-1,4-dihydro-5-pyrimidinecarboxylic acid




Methyl 4-(4-cyanophenyl)-6-methyl-2-(methylsulfanyl)-1-[3-(trifluoromethyl)-phenyl]-1,4-dihydro-5-pyrimidinecarboxylate (Example 8; 9.51 g, 21.4 mmol) is dissolved in 175 ml THF/ethanol/water (10:5:1). Potassium hydroxide (3.59 g, 64.1 mmol) is added, and the reaction mixture is stirred overnight. The reaction mixture is diluted with 100 ml of water, and the aqueous phase is washed with diethyl ether. The resulting aqueous phase is acidified to pH 1 with hydrochloric acid, and the resulting precipitate is filtered off, dissolved in ethyl acetate and washed with water. After drying with sodium sulfate, the solvent is removed in vacuo to give the product.
Yield: 655 mg (7% of th.) LC-MS (method 5): Rt=3.77 min. MS (ESIpos): m/z=432 (M+H)+1H-NMR (200 MHz, DMSO-d6): δ=12.32 (broad s, 1H); 7.52-7.98 (m, 8H); 5.75 (s, 1H); 2.15 (s, 3H); 2.02 (s, 3H) ppm.
Example 58
2-(Diethylamino)ethyl 4-(4-cyanophenyl)-6-methyl-2-(methylsulfanyl)-1-[3-(tri-fluoromethyl)phenyl]-1,4-dihydro-5-pyrimidinecarboxylate




4-(4-Cyanophenyl)-6-methyl-2-(methylsulfanyl)-1-[3-(trifluoromethyl)phenyl]-1,4-dihydro-5-pyrimidinecarboxylic acid (Example 57; 100 mg, 0.23 mmol) is dissolved in 3 ml acetone. Potassium carbonate (67 mg, 0.49 mmol), N-(2-bromoethyl)-N,N-diethylamine hydrobromide (67 mg, 0.25 mmol) and N,N,N-tributyl-1-butan-aminium iodide ( ̃5 mg) are added, and the reaction mixture is stirred at room temperature for 4 hours. The solvent is removed in vacuo, ethyl acetate is added and after extraction with 1 N sodium hydroxide solution, the organic layer is dried and the solvent is evaporated. The crude product is purified via preparative HPLC (RP18-column; eluent: acetonitrile-water, gradient 10:90 to 90:10).
Yield: 90 mg (73% of th.) LC-MS (method 5): Rt=3.00 min. MS (ESIpos): m/z=531 (M+H)+1H-NMR (200 MHz, DMSO-d6): δ=7.55-7.98 (m, 8H); 5.78 (s, 1H); 4.07 (t, 2H); 2.62 (t, 2H); 2.41-2.58 (m, 4H); 2.15 (s, 3H); 2.03 (s, 3H); 0.91 (t, 6H) ppm.
In analogy to the procedure of Example 58, the following compounds are prepared:

















Starting
Yield
Rt [min]
Mass


Ex.-No.
Structure
materials
[%]
(method)
[M + H]+




















59




Example 57; 1-(2-chloro- ethyl)- pyrrolidine hydrochloride
43
3.80 (4)
513


 


60




Example 57; ethyl bromo- acetate
42
4.50 (4)
518


 


61




Example 57; 2-chloro- N-ethyl- N-methyl- ethanamine hydrochloride
48
3.70 (4)
517


 


62




Example 57; 1-(2-chloro- ethyl)- piperidine hydrochloride
46
3.80 (4)
543


 


63




Example 57; methyl chloroacetate
71
4.10 (7)
504









C. Operative Examples Relating to Pharmaceutical Compositions

The compounds according to the invention can be converted into pharmaceutical preparations as follows:
Tablet:
Composition:
100 mg of the compound of Example 1, 50 mg of lactose (monohydrate), 50 mg of maize starch (native), 10 mg of polyvinylpyrrolidone (PVP 25) (from BASF, Ludwigshafen, Germany) and 2 mg of magnesium stearate. Tablet weight 212 mg, diameter 8 mm, curvature radius 12 mm.
Preparation:
The mixture of active component, lactose and starch is granulated with a 5% solution (m/m) of the PVP in water. After drying, the granules are mixed with magnesium stearate for 5 min. This mixture is moulded using a customary tablet press (tablet format, see above). The moulding force applied is typically 15 kN.
Orally Administrable Suspension:
Composition:
1000 mg of the compound of Example 1, 1000 mg of ethanol (96%), 400 mg of Rhodigel (xanthan gum from FMC, Pennsylvania, USA) and 99 g of water.
A single dose of 100 mg of the compound according to the invention is provided by 10 ml of oral suspension.
Preparation:
The Rhodigel is suspended in ethanol and the active component is added to the suspension. The water is added with stirring. Stirring is continued for about 6 h until the swelling of the Rhodigel is complete.

## Claims

1. A compound of formula (I-A)




wherein
A represents a phenyl ring,
R1 represents hydrogen, or C1-C6-alkyl wherein C1-C6-alkyl can be further substituted with one to three identical or different radicals selected from the group consisting of halogen, hydroxy and C1-C4-alkoxy,
R2 represents cyano,
R3 represents hydrogen,
R4 represents C1-C6-alkyl, C1-C6-alkylcarbonyl, C1-C6-alkoxycarbonyl, hydroxy-carbonyl, aminocarbonyl, mono- or di-C1-C4-alkylaminocarbonyl, C6-C10-aryl-aminocarbonyl, heteroarylcarbonyl, heterocyclylcarbonyl, heteroaryl, heterocyclyl or cyano, wherein C1-C6-alkyl, C1-C6-alkylcarbonyl, C1-C6-alkoxycarbonyl, mono- and di-C1-C4-alkylaminocarbonyl can be further substituted with one to three identical or different radicals selected from the group consisting of C3-C8-cycloalkyl, hydroxy, C1-C4-alkoxy, C1-C4-alkoxycarbonyl, hydroxycarbonyl, aminocarbonyl, mono- and di-C1-C4-alkylaminocarbonyl, C1-C4-alkylcarbonyl-amino, amino, mono- and di-C1-C4-alkylamino, heteroaryl, heterocyclyl, tri-(C1-C6-alkyl)-silyl and cyano,
R5 represents C1-C4-alkyl,
R6a represents hydrogen, C1-C6-alkylcarbonyl, C3-C8-cycloalkylcarbonyl, wherein C1-C6-alkyl-carbonyl can be substituted with one to three identical or different radicals selected from the group consisting of C3-C8-cycloalkyl, hydroxy, C1-C4-alkoxy, amino, mono- and di-C1-C4-alkylamino,
R7 represents halogen, or C1-C6-alkyl, wherein C1-C6-alkyl can be further substituted with one to three identical or different radicals selected from the group consisting of halogen, hydroxy and C1-C4-alkoxy, and
Y1, Y2, Y3 and Y4 each represent CH.

2. The compound of formula (I-A) according to claim 1, wherein
A represents a phenyl, ring,
R1 hydrogen, C1-C6-alkyl wherein C1-C6-alkyl can be further substituted with one to three identical or different radicals selected from the group consisting of halogen, hydroxy and C1-C4-alkoxy,
R2 represents cyano,
R3 represents hydrogen,
R4 represents C1-C6-alkylcarbonyl, C1-C6-alkoxycarbonyl, hydroxycarbonyl, amino-carbonyl, mono- or di-C1-C4-alkylaminocarbonyl, C6-C10-arylaminocarbonyl, heteroarylcarbonyl, heterocyclylcarbonyl, heteroaryl, heterocyclyl or cyano, wherein C1-C6-alkylcarbonyl, C1-C6-alkoxycarbonyl, mono- and di-C1-C4-alkylaminocarbonyl can be further substituted with one to three identical or different radicals selected from the group consisting of C3-C8-cycloalkyl, hydroxy, C1-C4-alkoxy, C1-C4-alkoxycarbonyl, hydroxycarbonyl, aminocarbonyl, mono- and di-C1-C4-alkylaminocarbonyl, C1-C4-alkylcarbonylamino, amino, mono- and di-C1-C4-alkylamino, heteroaryl, heterocyclyl and tri-(C1-C6-alkyl)-silyl,
R5 represents C1-C4-alkyl,
R6a represents hydrogen, C1-C6-alkylcarbonyl, C3-C8-cycloalkylcarbonyl, wherein C1-C6-alkyl-carbonyl, can be substituted with one to three identical or different radicals selected from the group consisting of C3-C8-cycloalkyl, hydroxy, C1-C4-alkoxy, amino, mono- and di-C1-C4-alkylamino,
R7 represents halogen, or C1-C6-alkyl, wherein C1-C6-alkyl can be further substituted with one to three identical or different radicals selected from the group consisting of halogen, hydroxy and C1-C4-alkoxy, and
Y1, Y2, Y3 and Y4 each represent CH.

3. The compound of formula (I-A) according to claim 1, wherein
A represents a phenyl ring,
R1 represents hydrogen, methyl, ethyl, trifluoromethyl or trifluoromethoxy,
R2 represents cyano,
R3 represents hydrogen,
R4 represents C1-C6-alkylcarbonyl, C1-C6-alkoxycarbonyl, hydroxycarbonyl, aminocarbonyl, mono- or di-C1-C4-alkylaminocarbonyl or cyano, wherein C1-C6-alkylcarbonyl, C1-C6-alkoxycarbonyl and mono-C1-C4-alkylaminocarbonyl can be substituted with one to three identical or different radicals selected from the group consisting of C3-C6-cycloalkyl, hydroxy, C1-C4-alkoxy, C1-C4-alkoxycarbonyl, amino, mono- or di-C1-C4-alkylamino, heteroaryl and heterocyclyl,
R5 represents methyl or ethyl,
R6a represents hydrogen, C1-C6-alkylcarbonyl or C3-C6-cycloalkylcarbonyl, wherein C1-C6-alkylcarbonyl can be substituted with a radical selected from the group consisting of C3-C6-cycloalkyl, hydroxy, C1-C4-alkoxy, amino, mono- and di-C1-C4-alkylamino,
R7 represents halogen, nitro, cyano, trifluoromethyl, trifluoromethoxy, methyl or ethyl, and
Y1, Y2, Y3 and Y4 each represent CH.

4. The compound of formula (I-A) according to claim 1, wherein
A represents a phenyl ring,
R1 and R3 each represent hydrogen,
R2 represents cyano,
R4 represents C1-C4-alkylcarbonyl or C1-C4-alkoxycarbonyl, wherein C1-C4-alkoxycarbonyl can be substituted with a radical selected from the group consisting of hydroxy, C1-C4-alkoxy, C1-C4-alkoxycarbonyl, mono- and di-C1-C4-alkylamino, heteroaryl and heterocyclyl,
R5 represents methyl,
R6a represents hydrogen, C1-C6-alkylcarbonyl or C3-C6-cycloalkylcarbonyl,
R7 represents trifluoromethyl or nitro, and
Y1, Y2, Y3 and Y4 each represent CH.

5. The compound of general formula (I-A) according to claim 1, wherein R1 is hydrogen.

6. The compound of formula (I-A) according to claim 1, wherein R4 is C1-C4-alkoxycarbonyl, which can be substituted with dimethylamino, diethylamino, N -ethylmethylamino, pyrrolidino or piperidino, or wherein R4 is C1-C4-alkylcarbonyl.

7. The compound of formula (I-A) according to claim 1, wherein R5 is methyl.

8. The compound of formula (I-A) according to claim 1, wherein R7 is trifluoromethyl or nitro.

9. The compound of formula (I-A) according to claim 1, wherein R6a is hydrogen.

10. A compound of formula (I-C)



wherein
Z represents CH, and R1, R3 and R4 have the meaning indicated in claim 1.

11. A composition containing at least one compound of formula (I-A) or (I-C), as defined in claims 1 or 10, and a pharmacologically acceptable diluent.