Quinazolin-4-one ampa antagonists

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DESCRIPTION

Quinazolin-4-ones as AMPA antagonists.

Background of the invention

The present invention relates to quinazolin-4-ones of formula I, as described below, their pharmaceutically acceptable salts, pharmaceutical compositions containing them and their use to treat neurodegenerative, psychotropic and drug and alcohol-induced disorders of the system. central and peripheral nervous.

The role of excitatory amino acids, such as glutamic acid and aspartic acid, as predominant mediators of excitatory synaptic transmission in the central nervous system has been well established. Watkins & Evans, Ann. Rev. Pharmacol. Toxicol., 21, 165 (1981); Monaghan, Bridges, and Cotman, Ann. Rev. Pharmacol. Toxicol., 29, 365 (1989); Watkins, Krogsgaard-Larsen, and Honore, Trans. Pharm. Sci., 11, 25 (1990). These amino acids function in synaptic transmission primarily through excitatory amino acid receptors. These amino acids also participate in various other physiological processes such as motor control, respiration, cardiovascular regulation, sensory perception, and cognition.

Excitatory amino acid receptors are classified into two general types. Receptors that are directly coupled to the opening of cation channels in the cell membrane of neurons are called "ionotropic." This type of receptor has been subdivided into at least three subtypes, which are defined by the depolarizing actions of the selective agonists N-methyl-D-aspartate (NMDA), a-amino-3-hydroxy-5-methylisoxazole4-propionic acid ( AMPA), and kainic acid (KA). The second general type is the G protein or second messenger-linked "metabotropic" excitatory amino acid receptor. This second group, when activated by the agonists quisqualate, ibotenate, or trans 1-aminocyclopentane-1,3-dicarboxylic acid, leads to enhanced hydrolysis of phosphoinositide in the postsynaptic cell. Both types of receptors appear to not only mediate normal synaptic transmission along excitatory pathways, but also participate in modifying the synaptic connection during development and changes in the efficiency of synaptic transmission throughout development. life. Schoepp, Bockaert, and Sladeczek. Trends in Pharmacol. Sci., 11, 508 (1990); McDonald and Johnson, Brain Research Reviews, 15, 41 (1990).

Excessive or inappropriate stimulation of excitatory amino acid receptors leads to neuron damage or loss by a mechanism known as excitotoxicity. This process has been suggested to mediate neuronal degeneration in various diseases. The medical consequences of such neuronal degeneration make the reduction of these degenerative neurological processes an important therapeutic goal.

Excitatory amino acid excitotoxicity has been implicated in the pathophysiology of various neurological disorders. This excitotoxicity has been implicated in the pathophysiology of acute and chronic neurodegenerative diseases including stroke, cerebral ischemia, spinal cord trauma, head trauma, Alzheimer's disease, Huntington's chorea, lateral amyotrophic sclerosis, epilepsy, AIDS-induced dementia, perinatal hypoxia. , hypoxia (such as conditions caused by strangulation, surgery, smoke inhalation, suffocation, immersion drowning, obstruction drowning, electrocution or drug or alcohol overdose), cardiac arrest, neuronal damage from hypoglycemia, eye damage and retinopathy, idiopathic and drug-induced Parkinson's disease, and brain deficiencies after heart bypass surgery and transplantation. Other neurological diseases, which are caused by glutamate dysfunction, require neuromodulation. These other neurological diseases include muscle spasms, headaches, urinary incontinence, psychosis, withdrawal from an addiction (such as alcoholism and drug addiction including addiction to opioids, cocaine, and nicotine), tolerance to opioids, anxiety, vomiting, brain edema, pain acute and chronic, seizures, retinal neuropathy, tinnitus and tardive dyskinesia. The use of a neuroprotective agent, such as an AMPA receptor antagonist, is believed to be useful in treating these disorders and / or in reducing the amount of neurological damage associated with these disorders. Excitatory amino acid receptor (EAA) antagonists are also useful as analgesic agents.

Various studies have shown that AMPA receptor antagonists are neuroprotective in models of focal and global ischemia. The competitive AMPA receptor antagonist NBQX (2,3-dihydroxy6-nitro-7-sulfamoylbenzo [f-] quinoxaline) has been reported to be effective in preventing global and focal ischemic damage. Sheardown et al., Science, 247, 571 (1900); Buchan et al., Neuroreport, 2, 473 (1991); LePeillet et al., Brain Research, 571, 115 (1992). The non-competitive AMPA receptor antagonist GKYI 52466 has been shown to be an effective neuroprotective agent in rat models of global cerebral ischemia. LaPeillet et al., Brain Research, 571, 115 (1992). These studies strongly suggest that delayed neuronal degeneration in cerebral ischemia involves glutamate excitotoxicity mediated at least in part by AMPA receptor activation. In this way, AMPA receptor antagonists may prove useful as neuroprotective agents and ameliorate the neurological consequences of cerebral ischemia in humans. Use of 3-substituted 2- (1H) quinolone derivatives, 3-substituted 4-hydroxy-2- (1H) quinolone derivatives, 2,4-dioxo-1,2,3,4 derivatives 3-substituted tetrahydro-3quinoline and quinazolin-2,4-diones as AMPA receptor antagonists are described in international patent application WO-A-93/11115, in European patents EP-A-0481676, EP -A-0459561 and in the international patent application WO-A-95/19346 respectively.

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Summary of the invention

The present invention relates to compounds of the formula:

<img file="ES2245015T3_D0001.tif" />

wherein A is a fused benzo or thiene aromatic ring;

B is phenyl, pyridyl, or pyrimidyl;

X is N or CH,

Y and Z, taken together, that is, YZ, are -CH₂NH- or -NHCH₂-;

R¹ is selected from hydrogen, alkyl (C₁-C₆) optionally substituted with one to three fluorine, cyano, halo, amino, nitro and alkoxy (C₁-C₆) optionally substituted with one to three fluorine atoms;

R² is halo, cyano, alkyl (C₁-C₆) optionally substituted with one to three fluorine, nitro, amino, alkylthio (C₁-C₆), alkoxy (C₁-C₆) optionally substituted with one to three fluorine atoms, hydroxy, HC (= O) -, alkyl (C₁-C₆) OC (= O) - or NH2-C (= O) -;

R³ and R⁴ are independently selected from hydrogen, (C1-C6) alkyl optionally substituted with one to three fluorine atoms, halo, cyano, hydroxy, (C1-C6) alkoxy optionally substituted with one to three fluorine atoms, -C (= O) H, -CH2 OR⁵ and -CH2NR⁶R⁷;

R⁵ is hydrogen, alkyl (C₁-C₆) or -C (= O) alkyl (C₁ -C₆); Y

R⁶ and R⁷ are independently selected from hydrogen, alkyl (C₁-C₆) and -C (= O) alkyl (C₁-C₆);

or R⁶ and R⁷, taken together with the nitrogen to which they are attached, form a four to seven membered saturated or unsaturated ring in which one of the carbon atoms of such a ring can optionally be replaced by oxygen or nitrogen (e.g. a morpholine, piperidine , pyrrolidine, piperizine, azetidine, pyrrole ring or oxazoline); and the pharmaceutically acceptable salts of such compounds.

The present invention also relates to the pharmaceutically acceptable acid addition salts of the compounds of formula I. The acids that are used to prepare the pharmaceutically acceptable acid addition salts of the base compounds of this invention mentioned above are those that form non-toxic acid addition salts, that is, salts containing pharmacologically acceptable anions, such as the salts. hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, acetate, lactate, citrate, acid citrate, tartrate, bitartrate, succinate, maleate, fumarate, gluconate, saccharate, benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate [ie, 1,1'-methylene-bis- (2-hydroxy-3-naphthoate)].

Examples of preferred compounds of formula I are those in which R¹ it is fluorine.

Other examples of preferred compounds of formula I are those in which YZ is -CH2NH-, ring A is a benzo ring and R¹ it is fluorine.

Other examples of preferred compounds of formula I are those in which R² it is halo, methyl or trifluoromethyl.

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Other examples of preferred compounds of formula I are those in which YZ is -CH₂NH-, ring A is a benzo ring, R¹ is fluoro, ring B is 2-pyridyl or phenyl and R³ is cyano, fluoro, methyl, or -CH₂NR⁶R⁷.

Other more specific embodiments of this invention are as follows:

(a) compounds of formula I in which ring A is benzo;

(b) compounds of formula I in which ring A is thiene;

(c) compounds of formula I in which ring B is phenyl;

(d) compounds of formula I in which ring B is a pyridine or pyrimidine ring; and (e) compounds of formula I in which R⁵ and R⁷Together with the nitrogen to which they are attached, they form a morpholine or pyrrolidine ring.

Examples of specific compounds of this invention are:

3- (2-chloro-phenyl) -6-fluoro-2 - [(pyridin-2-ylmethyl) -amino] -3H-quinazolin-4-one;

6-fluoro-3- (2-methyl-phenyl) -2 - [(pyridin-2-ylmethyl) -amino] -3H-quinazolin-4-one;

3- (2-chloro-phenyl) -6-fluoro-2 - [(2-fluorophenyl-methyl) -amino] -3H-quinazolin-4-one;

3- (2-chloro-phenyl) -2 - [(2-cyanophenyl-methyl) -amino] -6-fluoro-3H-quinazolin-4-one;

3- (2-chloro-phenyl) -2 - [(6-diethylaminomethylpyridin-2-ylmethyl) -amino] -6-fluoro-3H-quinazolin-4-one;

3- (2-chloro-phenyl) -6-fluoro-2 - [(6-pyrrolidin-1-ylmethyl-pyridin-2-ylmethyl) -amino) -3H-quinazolin-4-one;

3- (2-chloro-phenyl) -2 - [(3-pyrrolidin-1-ylmethyl-phenylamino) -methyl] -3H-thieno [3,2-d] pyrimidin-4-one;

3- (2-methyl-phenyl) -2 - [(3-pyrrolidin-1-ylmethyl-phenylamino) -methyl] -3H-thieno [3,2-d] pyrimidin-4-one;

3- (2-chloro-phenyl) -2 - [(2-fluoro-phenylamino) -methyl] -3H-thieno [3,2-d] pyrimidin-4-one;

3- (2-Chloro-pyrid-3-yl) -2 - [(3-pyrrolidin-1-ylmethyl-phenylamino) -methyl] -3H-thieno [3,2-d] pyrimidin-4-one;

2- {[3- (2-Chloro-pyrid-3-yl) -4-oxo-3,4-dihydro-thieno [3,2-d] pyrimidin-2-ylmethyl] -amino} -benzonitrile;

3- (2-chloro-phenyl) -2 - [(3-pyrrolidin-1-ylmethyl-phenylamino) -methyl] -3H-quinazolin-4-one;

6-chloro-3- (2-chloro-phenyl) -2 - [(3-pyrrolidin-1-ylmethyl-phenylamino) -methyl] -3H-quinazolin-4-one;

6-chloro-3- (2-chloro-phenyl) -2 - [(3-diethylaminomethyl-phenylamino) -methyl] -3H-quinazolin-4-one;

6-chloro-3- (2-chloro-pyrid-3-yl) -2 - [(3-diethylaminomethyl-phenylamino) -methyl] -3H-quinazolin-4-one;

6-chloro-3- (2-trifluoromethyl-phenyl) -2 - [(3-diethylaminomethyl-phenylamino) -methyl] -3H-quinazolin-4-one;

2 - {[3- (2-chloro-pyridin-3-iI) -4-oxo-3,4-dihydro-quinazolin-2-ylmethyl] -amino} -benzonitrile;

2 - {[3- (2-methyl-pyridin-3-iI) -4-oxo-3,4-dihydro-quinazolin-2-ylmethyl] -amino} -benzonitrile;

2 - {[6-fluoro-3- (2-methyl-phenyl) -4-oxo-3,4-dihydro-quinazolin-2-ylmethyl] -amino} -nicotinonitrile; Y

2 - {[3- (2-chloro-phenyl) -4-oxo-3,4-dihydro-quinazolin-2-ylmethyl] -amino} -nicotinonitrile.

This invention also relates to the use of a compound of formula I in the manufacture of a pharmaceutical composition for treating a disease selected from stroke, cerebral ischemia, spinal cord trauma, head trauma, Alzheimer's disease, Huntington's chorea, amyotrophic sclerosis. lateral, epilepsy, AIDS-induced dementia, muscle spasms, migraines, urinary incontinence, psychosis, seizures, perinatal hypoxia, hypoxia (such as conditions caused by strangulation, surgery, smoke inhalation, suffocation, drowning by immersion, drowning from obstruction, electrocution, or drug or alcohol overdose), cardiac arrest, neuronal damage from hypoglycemia, tolerance to opioids, addiction withdrawal (such as alcoholism and drug addiction, including addiction to opioids, cocaine and nicotine), eye damage, retinopathy, retinal neuropathy, tinnitus, Idiopathic and drug-induced Parkinson's disease, anxiety, vomiting, cerebral edema, chronic or acute pain, tardive dyskinesia and brain deficiencies after heart bypass surgery and transplantation, in a mammal,

ES 2 245 015 T3 comprising an amount of a compound of formula I and a pharmaceutically acceptable carrier.

This invention also relates to the use of a compound of formula I in the manufacture of a medicament for the treatment of a disease selected from stroke, cerebral ischemia, spinal cord trauma, head trauma, Alzheimer's disease, Huntington's chorea, sclerosis. lateral amyotrophic, epilepsy, AIDS-induced dementia, muscle spasms, migraines, urinary incontinence, psychosis, seizures, perinatal hypoxia, hypoxia (such as conditions caused by strangulation, surgery, smoke inhalation, suffocation, drowning by immersion, drowning from obstruction, electrocution, or drug or alcohol overdose), cardiac arrest, neuronal damage from hypoglycemia, tolerance to opioids, addiction withdrawal (such as alcoholism and drug addiction, including addiction to opioids, cocaine and nicotine), eye damage, retinopathy, retinal neuropathy, tinnitus, Idiopathic and drug-induced Parkinson's disease, anxiety, vomiting, brain edema, chronic or acute pain, tardive dyskinesia, and brain deficiencies after heart bypass surgery and transplantation, in a mammal.

This invention also relates to the use of a compound of formula I in the manufacture of a pharmaceutical composition for treating a disorder or disease, which treatment can be achieved or promoted by reducing or inhibiting glutamate neurotransmission in a mammal, wherein the composition comprises a compound of formula I, or a pharmaceutically effective salt thereof and a pharmaceutically acceptable carrier.

This invention also relates to the use of a compound of formula I in the manufacture of a medicament for the treatment of a disorder or disease, treatment or prevention that can be achieved or promoted by reducing or inhibiting glutamate neurotransmission in a mammal.

Unless otherwise indicated, alkyl groups referred to herein, as well as alkyl moieties of other groups referred to herein (eg alkoxy) may be linear or branched and may also be cyclic (eg, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl) or be linear or branched and contain cyclic moieties.

The term "treating", as used herein, refers to reversing, alleviating, inhibiting progression, or preventing the disorder or disease to which said term applies, or one or more symptoms of said disorder or disease. The term "treatment" as used herein refers to the act of treating, treating "as defined immediately above.

Unless otherwise indicated, the terms "halo" and "halogen", as used herein, refer to fluorine, bromine, chlorine or iodine.

The compounds of formula I can have chiral centers and therefore can exist in different enantiomeric and diastereomeric forms. This invention relates to all optical isomers and all stereoisomers of compounds of formula I and their mixtures, and to all pharmaceutical compositions and methods of treatment defined above that contain or employ them, respectively.

Due to the substituent on the carbon at the "2" position and the oxo atom on the carbon at the "4" position of the quinazolin-4-one of formula I, the ring attached to the nitrogen at the "3" position cannot rotate freely. This restricted rotation means that the compounds of formula I exist in two isomeric or atropoisomeric forms. These atropoisomers can separate.

This invention includes, for example, those stereiosomers of compounds of formula I that are atropoisomers. Atropoisomers are isomeric compounds that are chiral, that is, each isomer is not superimposable with its mirror image and the isomers, once separated, rotate the polarized light equally but in opposite directions. Atropoisomers are distinguished from enantiomers in that atropoisomers do not possess a single asymmetric atom. Such compounds are conformational isomers that occur when rotation about a single bond in molecules is hindered or severely slowed as a result of steric interactions with other parts of the molecule and the substituents at both ends of the single bond are asymmetric. A detailed report on atropoisomers can be found in Jerry March, Advanced Organic Chemistry, 101-102 (4^to ed. 1992) and in Oki, Top. Stereochem., 14, 1-81 (1983).

The following structure describes the atropoisomerism of the compound of formula I.

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<img file="ES2245015T3_D0002.tif" />

The bold lines in formula Ia indicate that the bonded atoms, and the groups attached to them, are sterically restricted to exist orthogonally above the plane of the quinazolinone ring. This steric restriction is due to a rotational energy barrier that prevents free rotation on the single bond that connects the nitrogen at the "3" position of the quinazolinone ring with the aryl group containing X (ie, phenyl or pyridyl).

Formulas I and Ia above include compounds identical to those described except for the fact that one or more hydrogens, carbons, or other atoms are replaced by their isotopes. Such compounds are useful as research and diagnostic tools in pharmacokinetic studies of metabolism and in binding assays. Specific research applications include radioligand binding assays, autoradiography studies, and in vivo binding studies.

Detailed description of the invention

Compounds of formula I can be prepared according to the methods of Scheme 1. In the Reaction Scheme and discussion that follows, unless otherwise indicated, rings A and B and substituents R¹ up to R⁷, Y and Z are as defined above for formula I.

Scheme 1

<img file="ES2245015T3_D0003.tif" />

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<img file="ES2245015T3_D0004.tif" />

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Scheme 3

<img file="ES2245015T3_D0005.tif" />

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Scheme 4

<img file="ES2245015T3_D0006.tif" />

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Scheme 5

<img file="ES2245015T3_D0007.tif" />

Scheme 1 illustrates methods for preparing intermediates of formula II, which can be converted to compounds of formula I. With respect to Scheme 1, a compound of formula V can be converted to an acetamide of formula IV by reacting it with acetyl chloride or acetic anhydride in the presence of a base in a reaction-inert solvent. Suitable solvents include methylene chloride, dimethoxyethane, t-butyl methyl ether, dichloroethane, tetrahydrofuran, and dioxane. Suitable bases include trialkylamines such as triethylamine and tributylamine, dimethylaminopyridine, and potassium carbonate, preferably triethylamine. The temperature of this reaction is in the range of from about 0 ° C to about 100 ° C for about 1 hour to about 10 hours, preferably from about 0 ° C to 30 ° C for about 3 hours.

Acetamide of formula IV can be cyclized to form a compound of formula III by reaction with an agent

ES 2 245 015 T3 dehydrating agent, in the presence of a catalyst, in a dry inert reaction solvent. Suitable dehydrating agents include acetic anhydride, phosphorous pentoxide, dicyclohexylcarbodiimide, and acetyl chloride, preferably acetic anhydride. Suitable catalysts include sodium or potassium acetate, acetic acid, p-toluenesulfonic acid, or boron trifluoride etherate, preferably sodium acetate. Suitable solvents include dioxane, toluene, diglyme or dichloroethane, preferably dioxane. The temperature of this reaction can range from about 0 ° C to about 150 ° C, and the reaction is generally carried out for about 1 hour to about 24 hours. The reaction is preferably carried out at about 80 ° C to 100 ° C, for about 3 to 10 hours.

Alternatively, the compound of formula V can be converted directly to a compound of formula III by reacting it with acetic anhydride in the presence of an acid catalyst in a solvent. Examples of acid catalysts that can be used are acetic acid, sulfuric acid, and p-toluenesulfonic acid. Acetic acid is preferred. Examples of solvents that can be used are toluene and xylene. Acetic acid is also the preferred solvent. The temperature for this reaction can range from about 20 ° C to about 150 ° C, and the reaction is generally carried out for about 10 minutes to about 10 hours. The reaction is preferably carried out at about 80 ° C to 120 ° C for about 2 to 5 hours.

The compound of formula III, formed by any of the above methods, can then be reacted with an amine of formula

<img file="ES2245015T3_D0008.tif" />

in a polar protic solvent, in the presence of an acid catalyst, to form a compound of formula II. Suitable acid catalysts include acetic acid, p-toluenesulfonic acid, and sulfuric acid, with acetic acid being preferred. Suitable polar protic solvents include acetic acid, methanol, ethanol, and isopropanol, with acetic acid being preferred. This reaction is generally carried out at a temperature of from about 20 ° C to about 150 ° C for about 1 hour to about 24 hours, preferably for about 6 hours at about 80 ° C to 120 ° C.

Alternatively, a compound of formula IV can be converted directly to a compound of formula II by reaction with a dehydrating agent, an amine of formula VIII, as described above, and a base, in an inert reaction solvent. Examples of dehydrating agents that can be used are phosphorous trichloride, phosphorous oxychloride, phosphorous pentachloride, and thionyl chloride, with phosphorous trichloride being preferred. Suitable bases include pyridine, lutidine, diisopropylethylamine, dimethylaminopyridine, triethylamine, and N-methyl morpholine. Suitable solvents include toluene, cyclohexane, benzene, and xylene. Pyridine is preferably used as the base and the reaction is carried out in toluene as the solvent. In some circumstances, when the combined reactants are a liquid, the reaction may proceed neat. The temperature can range from about 50 ° C to about 150 ° C, and the reaction is generally allowed to progress for about 1 hour to about 24 hours. It is preferably carried out at about 80 ° C to about 120 ° C for about 2-8 hours.

Scheme 2 illustrates the synthesis of compounds of formula I from the corresponding compounds of formula II. Regarding scheme 2, the reaction of a compound of II with the complex of dimethylformamide and dimethyl acetal (DMF-DMA) in dimethylformamide (DMF) at a temperature of approximately 50 ° C to approximately 180 ° C, preferably approximately 100 ° C at about 150 ° C it produces the corresponding amines of formula IX.

Aldehydes of formula X can be formed by reacting the corresponding enamines of formula IX with sodium periodate (NalO₄) or potassium permanganate (KMnO₄) in a solvent mixture containing water and an organic solvent such as ether, dimethoxyethane (DME), dioxane and tetrahydrofuran (THF), preferably THF, at a temperature of about 0 ° C to about 80 ° C, preferably about room temperature . An aqueous buffer is preferably added to the reaction mixture to maintain a pH of about 7.

The aldehydes formed in the above reaction can be converted to the corresponding compounds of formula I by reductive amination using, as amine, a compound of formula

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<img file="ES2245015T3_D0009.tif" />

The reductive amination can be carried out at temperatures in the range of about 0 ° C to about 150 ° C, preferably about 20 ° C to 100 ° C, using any of a variety of reducing agents, for example, sodium cyanoborohydride (NaBH₃CN), sodium triacetoxyborohydride (NaBH (OAc)₃), or sodium borohydride (NaBH₄), in a solvent such as methylene chloride, 1,2-dichloroethane, toluene, benzene, acetic acid, methanol, and ethanol. The preferred solvent will vary with the choice of reducing agent, as will be obvious to those skilled in the art. The reduction can also be achieved by hydrogenation, using hydrogen gas at a pressure of about 1 to about 5 atmospheres, a catalyst selected from rhodium, palladium, palladium hydroxide and platinum oxide. Hydrogenation can also be carried out using a chemical source of hydrogen such as ammonium formate or formic acid. Such transfer hydrogenation will use the same catalysts described above. The reductive amination can optionally be carried out in the presence of a dehydrating agent such as sodium sulfate, magnesium sulfate, calcium sulfate, or molecular sieves.

The reductive amination described above proceeds through an imine intermediate, as described in Scheme 3. If desired, the imine intermediate of formula XIII can be formed (and optionally isolated) by prehydrating the reaction mixture using an acid as acid. p-toluenesulfonic or sulfuric acid, with or without azeotropic removal of water, prior to the addition of the reducing agent. Treatment of the imine of formula XIII under any of the conditions previously described provides the corresponding compound of formula I.

Scheme 4 illustrates an alternative method of preparing the compounds of formula II from those of formula V. The compounds of formula II, thus formed, can then be converted to the desired compounds of formula I using the procedure illustrated in Scheme 2 and previously described. Referring to Scheme 4, a compound of formula V is reacted with a coupling reagent, an amine of formula VIII, as described above, and a base in an inert reaction solvent to form a compound of formula VI. Examples of suitable coupling reagents that activate carboxylic functionality are dicyclohexylcarbodiimide, N3-dimethylaminopropyl-N'-ethylcarbodiimide, 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ), carbonyldiimidazole (CDI), and diethylphosphoryl cyanide. Suitable bases include dimethylaminopyridine (DMAP) and triethylamine. Dimethylaminopyridine is preferred. A catalyst such as hydroxybenzotriazole (HBT) can also be used. The coupling is carried out in an inert solvent, preferably an aprotic solvent. Suitable solvents include acetonitrile, dichloromethane, dichloroethane, and dimethylformamide. The preferred solvent is dichloromethane. The mentioned reaction temperature is generally from about -30 to about 80 ° C, and is preferably from about 0 to about 25 ° C.

The compound of formula VI can be converted to a compound of formula VII by reaction with acetyl chloride or acetic anhydride in the presence of a base (for example, a trialkylamine such as triethylamine or tributylamine, dimethylaminopyridine or potassium carbonate) in a reaction-inert solvent. . Suitable solvents include methylene chloride, tetrahydrofuran, and chloroform, preferably methylene chloride. Preferably, triethylamine is used as the base. This reaction is generally carried out at a temperature of from about 0 ° C to about 35 ° C for about 1 hour to about 10 hours, preferably at about 30 ° C for about 3 hours.

The compound of formula VII is cyclized to a compound of formula II by reaction with triphenylphosphine, a base, and a dialkyl azodicarboxylate in a reaction-inert solvent. Examples of bases that can be used in this reaction are pyridine, triethylamine, and 4-dimethylaminopyridine, with 4-dimethylaminopyridine being preferred. Suitable solvents include dimethylformamide, tetrahydrofuran, and dioxane, with dioxane being preferred. Generally, this reaction is carried out at a temperature of about 25 ° C to about 125 ° C for about 1 hour to about 24 hours, preferably about 80 ° C to 120 ° C for about 8 to 15 hours.

Compounds of formula II can also be made according to the methods described in Miyashita, et al., Heterocycles, 42, 2, 691-699 (1996).

Scheme 5 illustrates a method for preparing compounds of formula I in which YZ is NHCH₂. Referring to Scheme 5, a 2-aminocarboxylic acid of formula V is reacted with an isothiocyanate of formula XIV to form the thione product of formula XV. This reaction is generally carried out in a solvent such as acetic acid, dioxane, tetrahydrofuran, chloroform, dichloroethane or benzene, preferably acetic acid, at a temperature of

ES 2 245 015 T3 at about 20 ° C to about 150 ° C, preferably at about the reflux temperature of the solvent, for about 0.25 hours to 24 hours, generally for about 1-6 hours.

The resulting thiones of formula XV can then be converted to the chlorinated compounds of formula XVI by reaction with a chlorinating agent such as phosphorus trichloride, phosphorus pentachloride, phosphorus oxychloride, thionyl chloride, sulfuryl chloride, or a mixture of one or more of these chlorinating agents, preferably a mixture of phosphorous oxychloride and phosphorous pentachloride. The reaction can be carried out without a solvent or in an inert solvent such as toluene, benzene, chloroform, dichloroethane, or dimethoxyethane. The reaction without solvent is preferred unless the reagents do not freely form a solution. This reaction is generally carried out at a temperature of about 20 ° C to about 180 ° C, preferably about 80 ° C to 150 ° C for about 0.5 to 24 hours, preferably about 1-6 hours.

The reaction of chlorides of formula XVI with an amine of formula

<img file="ES2245015T3_D0010.tif" />

produces the corresponding compounds of formula I in which YZ is NHCH₂. This reaction is generally carried out in a solvent such as methanol, ethanol, propanol, isopropanol, tetrahydrofuran, dioxane or dichloroethane, ethanol being preferred. The reaction is allowed to progress for about 1-24 hours, at a temperature of from about 20 ° C to about 150 ° C. Preferably, the reaction is allowed to progress for about 4-16 hours at a temperature of about 80 ° C to 120 ° C.

Unless otherwise indicated, the pressure of each of the above reactions is not critical. Generally, the reactions will be carried out at a pressure of about one to about three atmospheres, preferably atmospheric pressure (about one atmosphere).

Compounds of formula I which are basic in nature, are capable of forming a wide variety of different salts with different inorganic and organic acids. Although such salts must be pharmaceutically acceptable for administration to animals, in practice it is often desirable to initially isolate a compound of formula I from the reaction mixture as a pharmaceutically unacceptable salt and then simply convert it back to the free base compound. by treating with an alkaline reagent, and subsequently converting the free base to a pharmaceutically acceptable acid addition salt. The acid addition salts of the basic compounds of this invention are readily prepared by treating the basic compound with a substantially equivalent amount of the chosen mineral or organic acid in an aqueous solvent medium or in a suitable organic solvent such as methanol or ethanol. After careful evaporation of the solvent, the desired solid salt is obtained.

The acids that are used to prepare the pharmaceutically acceptable acid addition salts of the basic compounds of this invention are those that form non-toxic acid addition salts, that is, salts containing pharmacologically acceptable anions, such as hydrochloride salts, hydrobromide, hydroiodide, nitrate, sulfate or bisulfate, acid phosphate or phosphate, acetate, lactate, acid citrate or citrate, tartrate or bitartrate, succinate, maleate, fumarate, gluconate, saccharate, benzoate, methanesulfonate, and pamoate [ie, 1,1'-methylene-bis- (2-hydroxy-3-naphthoate)].

The compounds of formula I and their pharmaceutically acceptable salts (hereinafter also referred to as the active compounds of the invention) are useful for the treatment of neurodegenerative, psychotropic and alcohol- and drug-induced disorders of the central and peripheral nervous system and are potent AMPA receptor antagonists. The active compounds of the invention can therefore be used in the treatment or prevention of stroke, cerebral ischemia, spinal cord trauma, head trauma, Alzheimer's disease, Huntington's chorea, amyotrophic lateral sclerosis, epilepsy, AIDS-induced dementia, muscle spasms, headaches, urinary incontinence, psychosis, seizures, perinatal hypoxia, hypoxia (such as conditions caused by strangulation, surgery, smoke inhalation, choking, drowning from immersion, choking from obstruction, electrocution or drug or alcohol overdose), cardiac arrest, neuronal damage from hypoglycemia, tolerance to opioids, withdrawal from addictions (such as alcoholism and drug addiction, including addiction to opioids, cocaine and nicotine ), eye damage, retinopathy, retinal neuropathy, tinnitus, idiopathic and drug-induced Parkinson's disease, anxiety, vomiting, brain edema, chronic or acute pain, Tardive dyskinesia and brain deficiencies after heart bypass surgery and transplantation.

ES 2 245 015 T3

The in vitro and in vivo activity of the compounds of the invention for AMPA receptor antagonism can be determined by methods available to those skilled in the art. One method of determining the activity of the compounds of the invention is by inhibiting the sudden onset diseases induced by pentylenetetrazole (PTZ). Another method to determine the activity of the compounds of the invention is by blocking the absorption of⁴⁵AC²⁺ induced by AMPA receptor activation.

A specific method for determining the inhibition of pentylenetetrazole (PTZ) -induced sudden onset diseases is as follows. The activity of the compounds of the invention for the inhibition of pentylenetetrazole (PTZ) -induced sudden onset diseases in mice can be determined according to the following procedure. This assay examines the ability of compounds to block sudden onset and fatal diseases caused by PTZ. The measures taken are latency to clonic and tonic sudden onset diseases and death. The IDs₅₀s are determined based on the percentage of protection.

Male Charles River CD-1 mice, weighing 14-16 g on arrival and 25-35 g at the time of testing, served as subjects for these experiments. Thirteen mice per cage were housed under standard laboratory conditions on a L: D light cycle (Light: Dark) / 7 am: 7 pm for at least 7 days prior to experimentation. Food and water were available ad libitum until the time of testing.

All compounds were administered in a volume of 10 ml / kg. Drug carriers will depend on the solubility of the compound, but screening will generally be done using saline, distilled water, or E: D: S / 5: 5: 90 (5% emulsifier, 5% emulsifier). DMSO, and 90% saline) as an injection vehicle.

Mice were administered the test compounds or vehicle (ip, sc, or po) and plexiglass cages were placed in groups of five. At a predetermined time after these injections, the mice were given an injection of PTZ (ip, 120 mg / kg) and individual plexiglass cages were placed. The measurements that were taken during this five minute test period were: (1) latency to clonic sudden onset diseases, (2) latency to tonic sudden onset diseases, and (3) latency to death. The treatment groups were compared with the vehicle treated group using the Kruskal-Wallis, Anova and Mann-Whitney U tests (Statview). The percentage of protection was calculated for each group (number of subjects showing no sudden onset illness or death as indicated by a 300 second result) for each measure. ID50's were determined by Probit analysis (Biostat).

Another method to determine the activity of the compounds is to determine the effect of the compounds on the motor coordination of the mice. This activity can be determined according to the following procedure.

Male Charles River CD-1 mice, weighing 14-16 g at arrival and 23-35 g at the time of testing, served as subjects for these experiments. 13 mice per cage were housed under standard laboratory conditions on a L: D / 7 am: 7 pm light cycle for at least 7 days prior to experimentation. Food and water are available ad libitum until the time of the test.

All compounds were administered in a volume of 10 ml / kg. Drug carriers will depend on the solubility of the compound, but screening will generally be done using saline, distilled water, or E: D: S / 5: 5: 90 (5% emulsifier, 5% emulsifier). DMSO, and 90% saline) as an injection vehicle.

The equipment used in these studies consisted of a group of five 13.34 x 13.34 cm squares of wire mesh suspended on 11.43 cm steel poles connected to a 165.1 cm pole that was raised 38.1 cm above the laboratory table. These squares of wire mesh could be turned inside out.

Mice were administered the test compounds or vehicle (ip, sc, or po) and plexiglass cages were placed in groups of five. At a predetermined time after these injections, the mice were placed on top of the wire mesh squares and raised so that they were suspended upside down. During the test minute, mice were scored as 0 if they fell off the sieve, 1 if they stood upside down, or 2 if they climbed to the top. Treatment groups were compared with the vehicle-treated group using the Kruskal-Wallis and Mann-Whitney U tests (Statview).

The following describes a specific method to determine absorption blockage of the ⁴⁵AC²+ induced by AMPA receptor activation.

Primary neuronal cultures

Primary cultures of rat cerebellar granule neurons were prepared as described by Parks, TN, Artman, LD, Alasti, N., and Nemeth, EF, Modulation Of N-Methil-D-Aspartate Receptor-Mediated Increases In Cytosolic Calcium In Cultured Rat Cerebellar Granule Cells, Brain Res. 552, 13-22 (1991). According to this method, the cerebellum of 8-day-old CD rats was excised, minced into 1-mm pieces, and incubated for 15 minutes at 37 ° C in a calcium and magnesium-free Tyrode solution containing 0 trypsin. 1 %. The tissue was then ground using a fine-tipped Pasteur pipet. The cell suspension was placed in poly-D-lysin coated 96-well tissue culture plates at 10⁵ cells per well. The medium consisted of Minimal Essential Medium (MEM), with Earle's salts, 10% heat inactivated Fetal Bovine Serum, 2 mM L-glutamine, 21 mM glucose, Penicillin-Streptomycin.

ES 2 245 015 T3 (100 units per ml) and 25 mM KCl. After 24 hours, the medium was replaced with fresh medium containing 10 µΜ cytosine arabinoside to inhibit cell division. Cultures should be used at 6-8 DIV.

Absorption of⁴⁵ AC² induced by AMPA receptor activation

The effects of drugs on the absorption of ⁴⁵AC²⁺ induced by AMPA receptor activation can be analyzed in rat cerebellar granule cell cultures. The 96-well plate cultures were pre-incubated for approximately 3 hours in serum-free medium and then for 10 minutes in a Mg-free balanced salt solution.²+ (in mM: 120 NaCl, 5 KCI, 0.33 NaH2PO4, 1.8 CaCl2, 22.0 glucose and 10.0 HEPES at pH 7.4) containing DTT 0.5 mM, 10 μM glycine and drugs at 2X final concentration. The reaction was started by rapid addition of an equal volume of balanced salt solution containing 100 μM of AMPA receptor agonist kainic acid and⁴⁵AC²+ (final specific activity 250 Ci / mmol). After 10 minutes at 25 ° C, the reaction was stopped by aspirating the solution containing⁴⁵AC²⁺ and washing cells 5X in an ice cold balanced salt solution containing no added calcium and containing 0.5 mM EDTA. The cells were then lysed by incubation overnight in 0.1% Triton-X100 and the radioactivity in the lysate was then determined. All compounds of the invention, which were tested, had IC50s of less than 5 µM.

The compositions of the present invention can be formulated in a traditional manner using one or more pharmaceutically acceptable excipients. Thus, the active compounds of the invention can be formulated for oral, sublingual, transcutaneous (eg, patch) intranasal, parenteral (eg, intravenous, intramuscular, or subcutaneous) or rectal administration or in a suitable form of administration by inhalation or insufflation.

For oral administration, the pharmaceutical compositions may take the form of, for example, tablets or capsules prepared by conventional methods with pharmaceutically acceptable excipients such as binding agents (for example, pregelatinized cornstarch, polyvinylpyrrolidone, or hydroxypropylmethylcellulose); fillers (eg, lactose, microcrystalline cellulose, or calcium phosphate); lubricants (eg, magnesium stearate, talc, or silica); disintegrants (eg potato starch or sodium starch glycolate); or wetting agents (eg, sodium lauryl sulfate). Tablets can be coated by methods well known in the art. Liquid preparations for oral administration can take the form of, for example, solutions, syrups or suspensions or they can be presented as a dry product for constitution with water or other suitable vehicle before use. Such liquid preparations can be prepared by conventional means with pharmaceutically acceptable additives such as anti-settling agents (eg, sorbitol syrup, methylcellulose, or edible hydrogenated fats); emulsifying agents (eg, lecithin or acacia); non-aqueous vehicles (eg, almond oil, oily esters, or ethyl alcohol); and preservatives (eg, methyl or propyl p-hydroxybenzoates or sorbic acid).

Compositions for sublingual administration may take the form of tablets or lozenges formulated in a conventional manner.

The active compounds of the invention can be formulated for parenteral administration by injection, which includes using traditional catheterization techniques or intravenous infusion. Formulations for injection may be presented in unit dose form, for example, in ampoules or in multi-dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as anti-settling, stabilizing, and / or dispersing agents. Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, eg, sterile pyrogen-free water, before use.

The active compounds of the invention can also be formulated in rectal compositions such as suppositories or retention enemas, for example, containing conventional suppository bases such as cocoa butter or other glycerides.

For intranasal administration or administration by inhalation, the active compounds of the invention are conveniently administered in the form of a solution or suspension from a container provided with a mechanical spray for spraying that is pressed or pumped by the patient or as an aerosol presentation from a container. pressurized or a nebulizer, with the use of a suitable propellant, for example, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve to dispense a metered amount. The pressurized container or nebulizer may contain a solution or suspension of the active compound. Capsules and cartridges (made, for example, of gelatin) for use in an inhaler or insufflator can be formulated containing a powder mixture of a compound of the invention and a suitable powder base such as lactose or starch.

A proposed dose of the active compounds of the invention for oral, parenteral or sublingual administration to the average adult human for the treatment of the aforementioned diseases (eg, stroke) is 0.01 to 50 mg / kg of the active ingredient per dose. unit that could be administered, for example, 1 to 4 times a day.

Aerosol formulations for the treatment of the aforementioned diseases (eg, stroke) in the average adult human are preferably arranged so that each metered dose or "instantaneous discharge"

ES 2 245 015 T3 "aerosol" contains from 20 pg to 1,000 pg of the compound of the invention. The total daily dose with an aerosol will be within the range of 100 pg to 10 mg. Administration can be several times a day, for example 2, 3, 4 or 8 times, giving for example 1, 2 or 3 doses each time.

The following Examples illustrate the preparation of compounds of the present invention. Commercial reagents were used without further purification. Melting points are uncorrected. All NMR data was recorded at 250, 300 or 400 MHz in deuterated chloroform unless otherwise indicated and reported in parts per million (δ) and referenced to the fixed deuterium signal of the solvent sample. All non-aqueous reactions were run in dry glass vessels with dry solvents under an inert atmosphere for convenience and to maximize yields. All reactions were stirred with a magnetic stir bar unless otherwise indicated. Unless otherwise indicated, all mass spectra were obtained using chemical impact conditions. Ambient temperature refers to 20-25 ° C. Melting points are uncorrected.

Example 1

2 - {[3- (2-chloro-pyridin-3-yl) -6-fluoro-4-oxo-3,4-dihydro-quinazolin-2-ylmethyl] -amino} -benzonitrile

A mixture of 3- (2-chloro-pyridin-3-yl) -6-fluoro-3,4-dihydro-quinazolin-4-one-2-carboxaldehyde hydrate (0.164 g, 0.51 mmol), toluene ( 25 mL), anthranilonitrile (0.135 g, 1.12 mmol), and glacial acetic acid (0.064 mL, 1.12 mmol) were refluxed for 6 hours with azeotropic removal of water. The mixture was cooled to room temperature and anthranilonitrile (0.135 g, 1.12 mmol) and glacial acetic acid (1 mL) were added. The reaction was refluxed for 24 hours. P-Toluenesulfonic acid was added in catalytic amounts (a couple mg) and the reaction was refluxed for an additional 24 hours. The mixture was cooled to room temperature, diluted with ethyl acetate, and extracted with saturated aqueous bicarbonate, water, and brine. The organic layer was dried over sodium sulfate and concentrated to give 0.378 g of the crude imine as a dark oil. The imine had an R_F 0.15 on silica gel TLC developed with 30% ethyl acetate / hexane, UV detection.

The crude imine (0.378 g) was dissolved in ethanol (10 mL) and 10% palladium on carbon (0.14 g) and formic acid (0.5 mL) were added. The mixture was stirred at room temperature overnight. The mixture was filtered through celite and the filtrate was concentrated. The residue was taken up in 1 mL of 50% ethyl acetate / hexane and applied to a column of silica gel (1x4 inches (2.5 x 10.2 cm), packed with 25% ethyl acetate / hexane ) for purification by flash chromatography. The elution was carried out as follows: 25% ethyl acetate / hexane (50 mL), unweighted recovered anthranilonitrile (50 mL), traces of impurities (250 mL), and 0.112 g (54%) of

2- {[3- (2-chloro-pyridin-3-yl) -6-fluoro-4-oxo-3,4-dihydro-quinazolin-2-ylmethyl] -amino} -benzonitrile as an oil which solidified leaving it to be y had: mp 183-187 ° C; ¹H NMR δ 8.66 (dd, J = 1.8, 4.8 Hz, 1H), 7.95-7.86 (m, 2H), 7.79 (dd, J = 1.8, 7, 8 Hz, 1H), 7.60-7.53 (m, 2H), 7.43 (dd, J = 1.5, 7.7 Hz, 1H), 7.33 (t, J = 8 Hz, 1H), 6.73 (t, J = 7 Hz, 1H), 6.42 (d, J = 8.4 Hz, 1H), 6.02 (s broad; 1H), 3.95 (c AB, Av₁-₃ = 4.5Hz, J = 2Hz, 2H); APCI MS m / z = 406 (P +¹).

Example 2

3- {[3- (2-chloro-phenyl) -6-fluoro-4-oxo-3,4-dihydro-quinazolin-2-ylmethyl] -amino} -benzonitrile

A mixture of 3- (2-chloro-phenyl) -6-fluoro-3,4-dihydro-quinazolin-4-one-2-carboxaldehyde (0.150 g, 0.50 mmol), glacial acetic acid (10 mL), 3-aminobenzonitrile (0.050 g, 0.42 mmol), and anhydrous sodium sulfate (0.71 g, 5 mmol) was stirred at room temperature overnight. TLC indicated that the imine had formed (Rf = 0.43 on silica gel TLC developed with 30% ethyl acetate / hexane, UV detection). Sodium triacetoxyborohydride (0.267 g, 1.26 mmol) was added and the reaction was allowed to stir over the weekend (72 hours). The mixture was poured into saturated aqueous bicarbonate and repeatedly extracted with ethyl acetate. The combined organic layer was dried over magnesium sulfate and concentrated to give a yellow solid. This solid was triturated with 50% ethyl ether / isopropyl ether to give 0.133 g (78%) of 3 - {[3- (2-chloro-phenyl) -6-fluoro-4-oxo-3,4-dihydro- quinazolin-2-ylmethyl] amino} -benzonitrile as an off-white solid having: mp 225-228 ° C; Ή NMR δ 7.93 (dd, J = 3.8 Hz, 1H), 7.87 (dd, J = 4.5, 8.5 Hz, 1H), 7.70 (dd, J = 1.5 , 7.5 Hz, 1H), 7.62-7.52 (m, 4H), 7.40 (dd, J = 1.5, 7.5 Hz, 1H), 7.20 (t, J = 8Hz, 1H), 6.97 (d, J = 7.5Hz, 1H), 6.80 (dd, J = 2.8Hz, 1H), 6.66 (s, 1H), 3, 88 (c AB, Avi-₃ = 39.5 Hz, J = 17 Hz, 2H). Analysis calculated for C₂₂H₁₄CIFN₄O-0.5H₂O: C, 63.85; H, 3.65; N, 13.54. Found: C, 63.90; H, 3.31; N, 13.46.

Example 3

3- (2-chlorO-phenyl) -2 - [(3-diethylaminomethyl-phenylamino) -methyl] -6-fluoro-3H-quinazolin-4-one

A mixture of 3- (N, N-diethylamino-methyl) -aniline (0.080 g, 0.45 mmol) and 3- (2-chloro-phenyl) -6-fluoro-3,4-dihydroquinazolin-4-one- 2-carboxaldehyde (0.150 g, 0.50 mmol) in methanol (15 mL) was refluxed for 48 hours to form the imine intermediate. The reaction was cooled to room temperature and sodium borohydride (0.038 g, 1 mmol) was added and the mixture was refluxed for 24 hours. More sodium borohydride (0.038 g, 1 mmol) was added and refluxing continued for 2 hours. Sodium borohydride (0.038 g, 1 mmol) was added and the mixture was refluxed for a further 18 hours. The reaction was allowed to stir overnight at room temperature. Water was added and the mixture was stirred for

ES 2 245 015 T3 minutes. The reaction mixture was repeatedly extracted with ethyl acetate. The combined organic phase was washed with saturated aqueous bicarbonate and brine. The organic phase was dried over magnesium sulfate and concentrated to give a dark yellow oil. The oil was flash chromatographed on silica gel (30 g), elution proceeding as follows: 20% ethyl acetate / hexane (150 mL), nothing; (50 mL), 0.015 g of an unknown product; (75 mL), nothing; (125 mL), 0.01 g, 3- (2-chloro-phenyl) -6-fluoro-3,4-dihydro-quinazolin-4-one-2-methanol; (350 mL), nothing; 30% ethyl acetate / hexane (300 mL), nothing; 50% ethyl acetate / hexane (450 mL), nothing; (350 mL) and chloroform (50 mL), 0.016 g (0.8%) of 3- (2-chloro-phenyl) -2 - [(3-diethylaminomethyl-phenylamino) -methyl] -6-fluoro-3H- quinazolin-4one as a viscous yellow oil having: 'II NmR δ 7.92 (dd, J = 3.8 Hz, 1H), 7.82 (dd, J = 5.9 Hz, 1H), 7.667.65 (m, 1H), 7.57-7.46 (m, 4H), 7.14 (t, J = 8Hz, 1H), 7.08 (s, 1H), 6.70 (d, J = 7Hz, 1H), 6.59 (d, J = 8Hz, 1H), 5.28 (s, 1H), 4.06-3.81 (m, 4H), 3.46 (q, J = 7Hz, 4H), 1.19 (t, J = 7Hz, 6H); APCI MS m / z = 465.1 (P +¹).

Example 4

3- (2-chlorO-phenyl) -6-fluorO-2- (pyrimidin-2-ylaminomethyl) -3H-quinazolin-4-one

A mixture of 3- (2-chloro-phenyl) -6-fluoro-3,4-dihydro-quinazolin-4-one-2-carboxaldehyde (0.200 g, 0.66 mmol), methanol (20 mL), and 2 -aminopyrimidine (0.059 g, 0.62 mmol) was refluxed for 29 hours. The mixture was cooled to room temperature and 10% palladium on carbon (0.236 g) and formic acid (1.1 mL) were added. The mixture was left stirring overnight at room temperature. The reaction was treated with 6N sodium hydroxide to adjust the pH to 10. The mixture was filtered through celite and the bed was washed with ethyl acetate. The filtrate was dried over magnesium sulfate and concentrated. The residue was flash chromatographed on silica gel (20 g), elution proceeding as follows: 10% ethyl acetate / hexane (500 mL), nothing; 20% ethyl acetate / hexane (250 mL), unweighted impurity, 20% ethyl acetate / hexane (100 mL), mixed fraction, unweighted; 20% ethyl acetate / hexane (150 mL) and 30% ethyl acetate / hexane (150 mL), 3- (2-chloro-phenyl) -6-fluoro-3,4-dihydroquinazolin-4-one- Unweighted 2-methanol (a by-product). Elution was continued: 30% ethyl acetate / hexane (410 mL), nothing; 40% ethyl acetate / hexane (450 mL), nothing; 40% ethyl acetate / hexane (50 mL) and 50% ethyl acetate / hexane (450 mL), 0.028 g (8%) of 3- (2-chloro-phenyl) -6-fluoro-2- ( pyrimidin-2-ylaminomethyl) 3H-quinazolin-4-one as a white solid having: mp 182-185 ° C;¹H NMR δ 8.29 (d, J = 5 Hz, 2H), 7.92 (dd, J = 3.8 Hz, 1H), 7.82 (dd, J = 5.9 Hz, 1H), 7 , 66-7.64 (m, 1H), 7.55-7.49 (m, 3H), 7.40-7.38 (m, 1H), 6.63 (t, J = 5Hz, 1H ), 4.28 (dd, J = 4.5, 18 Hz, 1H), 4.11 (dd, J = 4.18 Hz, 1H). Analysis calculated for C1₉H_nCIFN₅O.0.75 H₂O: C, 58.05; H, 3.20; N, 17.81. Found: C, 58.06; H, 3.25; N, 17.33. APCI MS m / z = 382 (P +¹).

Example 5

3- (2-chloro-pyridin-3-yl) -6-fluoro-2- (m-tolylamino-methyl) -3H-quinazolin-4-one

A mixture of 3- (2-chloro-pyridin-3-yl) -6-fluoro-3,4-dihydro-quinazolin-4-one-2-carboxaldehyde 6 (0.150 g, 0.49 mmol) and 3-diethylaminomethylaniline (0.073 g, 0.41 mmol) in methanol (10 mL) was stirred at room temperature for 72 hours and then concentrated to give 0.252 g of yellow solid imine having: APCI MS m / z = 464 (P +¹). The crude imine was dissolved in ethanol (10 mL) and 10% palladium on carbon (0.252 g) and formic acid (0.90 mL, 24.3 mmol) were added. The mixture was stirred at room temperature for 6 hours. Solid potassium carbonate (0.5 g) was added, the mixture was stirred 1 hour more. The reaction was filtered through celite and the bed was washed with ethyl acetate. The filtrate was concentrated. The residue was taken up in ethyl acetate and washed with saturated bicarbonate and brine, dried over magnesium sulfate, and concentrated to give a yellow oil. The oil was flash chromatographed on silica gel (15 g), elution proceeding as follows: 10% ethyl acetate / hexane (300 mL), nothing; 20% ethyl acetate / hexane (400 mL), nothing; 20% ethyl acetate / hexane (150 mL), 0.033 g (20%) of 3- (2-chloropyridin-3-yl) -6-fluoro-2- (m-tolylamino-methyl) -3H-quinazolin- 4-one as a white solid having: mp 177-180 ° C;¹H NMR δ 8.63 (dd, J = 2.5 Hz, 1H), 7.92 (dd, J = 3.8 Hz, 1H), 7.83 (dd, J = 5.9 Hz, 1H) , 7.76 (dd, J = 2.8Hz, 1H), 7.58-7.50 (m, 2H), 7.05 (t, J = 8Hz, 1H), 6.61 (d, J = 7 Hz, 1H), 6.43-6.39 (m, 2H), 3.90 (c AB, Av ^ 23 Hz, J = 7 Hz, 2H), 2.25 (s, 3 H) . Analysis calculated for C₂₁H₁₆CIFN₄O-0.25 H₂O: C; 63.16; H, 4.16; N, 14.03. Found: C, 63.06; H, 4.28; N, 13.72.

Example 6

3- (2-chloro-pyridin-3-yl) -6-fluoro-2 - [(6-methyl-pyridin-2-ylamino) -methyl] -3H-quinazolin-4-one

To a mixture of 3- (2-chloro-pyridin-3-yl) -6-fluoro-3,4-dihydro-quinazolin-4-one-2-carboxaldahyde (0.20 g, 0.66 mmol) and 2 -amino-6-methylpyridine (0.0476 g, 0.44 mmol) in methanol (10 mL) was added acetic anhydride (0.042 mL, 0.44 mmol). The mixture was gently refluxed overnight, cooled to room temperature, and diluted with saturated aqueous bicarbonate. This aqueous mixture was repeatedly extracted with ethyl acetate. The combined organic phase was washed with saturated aqueous bicarbonate and brine, dried over magnesium sulfate, and concentrated to give 0.303 g of crude imine as a yellow oil. The imine was dissolved in ethanol (10 mL) and 10% palladium on charcoal (0.17 g) and formic acid (0.75 mL) were added. The mixture was stirred at room temperature overnight. Sodium bicarbonate (0.5 g) was added. The mixture was stirred for a further 1 hour, then filtered through celite and the bed was washed with ethanol and ethyl acetate. The filtrate was concentrated to give a colorless solid. This solid was triturated with ethyl acetate getting the product to dissolve in the solvent. The solution was concentrated and flash chromatographed on silica gel (20 g). Elution proceeded as follows: 50% ethyl acetate / hexane (350 mL), nothing; 50% ethyl acetate / hexane (550 mL), a mixture of the desired product and 3- (217

ES 2 245 015 T3 chloro-pyrid-3-yl) -6-fluoro-3,4-dihydro-quinazolin-4-one-2-methanol as a white solid. This mixture was triturated with 50% isopropyl ether / hexane to give 0.025 g (14%) of 3- (2-chloro-pyridin-3-yl) -6-fluoro-2 - [(6-methyl-pyridin-2ylamino ) -methyl] -3H-quinazolin-4-one as a white solid having: mp 190-195 ° C;¹H NMR δ 8.56 (dd, J = 1.5 Hz, 1H), 7.91 (dd, J = 3.8Hz, 1H), 7.80-7.76 (m, 2H), 7.53 (dt, J = 3.8 Hz, 1H), 7.46 (dd, J = 5.8 Hz, 1H), 7.29 (t, J = 8 Hz, 1H), 6.44 (d, J = 7Hz, 1H), 6.24 (d, J = 8Hz, 1H), 5.64 (s, 1H), 4.27 (dd, J = 6.17Hz, 1H), 4.10 ( dd, J = 5.17Hz, 1H), 2.03 (s, 3H). Analysis calculated for C₂₀H₁₅CIFN₅Oh₂O: C, 58.05; H, 4.14; N, 16.92. Found: C, 58.45; H, 3.71; N, 16.53.

Example 7

3- (2-Chloro-phenyl) -6-fluoro-2- (pyridin-2-ylaminomethyl) -3H-quinazolin-4-one hydrochloride

A mixture of 3- (2-chloro-phenyl) -6-fluoro-3,4-dihydro-quinazolin-4-one-2-carboxaldehyde (0.20 g, 0.66 mmol), 2-aminopyridine (0.069 g, 0 , 73 mmol), and acetic acid (0.075 mL, 1.32 mmol) in toluene (25 mL) was refluxed for 5 hours with azeotropic removal of water. The reaction was cooled to room temperature and extracted with saturated aqueous bicarbonate. The phases were separated and the organic phase was dried over sodium sulfate and concentrated to give the crude imine as a brown foam having: mp 158-162 ° C. A part of this imine (0.10 g, 0.264 mmol) was dissolved in ethanol (10 mL) and 10% palladium on carbon (0.10 g) and formic acid (0.45 mL, 11.9 mmol) were added ). The mixture was stirred for 4 hours at room temperature, and then filtered through celite and the bed was rinsed with ethyl acetate and water. The layers of the two filtered phases were separated and the organic layer was washed with saturated aqueous bicarbonate, dried over magnesium sulfate and concentrated to give 0.065 g (65%) of the free base of the product as a brown oil having: ¹H NMR δ 8.02 (d, J = 4 Hz, 1H), 7.94 (dd, J = 5.9 Hz, 1H), 7.66-7.61 (m, 1H), 7.577.36 ( m, 6H), 6.58 (t, J = 5Hz, 1H), 6.50 (d, J = 8Hz, 1H), 5.83 (s broad, 1H), 4.25 (dd, J = 5.18 Hz, 1H), 4.03 (dd, J = 4.18 Hz, 1H); APCI MS m / z = 381.1 (P +¹). The free base was dissolved in ether (15 mL) and cooled to 0 ° C. Ethereal hydrochloric acid solution (0.20 mL, 0.2 mmol, approximately 1 N) was added to the solution. The precipitate was collected and dried to give 0.025 g of 3- (2-chloro-phenyl) -6-fluoro-2- (pyridin-2-ylaminomethyl) -3Hquinazolin-4-one hydrochloride as an amorphous solid having: mp 90-100 ° C.

Example 8

3- (2-chloro-pyridin-3-yl) -6-fluoro-2 - [(3-pyrrolidin-1-ylmethyl-phenylamino) -methyl] -3H-quinazolin-4-one

A mixture of 3- (2-chloro-pyridin-3-yl) -6-fluoro-3,4-dihydro-quinazolin-4-one-2-carboxaldehyde (0.150 g, 0.49 mmol), 3-pyrrolidin- 1-ylmethylaniline (0.044 g, 0.25 mmol), and acetic acid (0.14 mL, 2.45 mmol) in dichloroethane (10 mL) was cooled to 0 ° C and sodium triacetoxyborohydride (0.312 g, 1 , 47 mmol). The reaction was stirred for 1 hour at 0 ° C and then allowed to warm to room temperature. The reaction was stopped by the addition of saturated aqueous bicarbonate and stirring was continued for 30 minutes. The phases were separated and the organic layer was extracted with methylene chloride. The combined organic phase was washed with brine, dried over magnesium sulfate, and concentrated. The residue was subjected to flash chromatography on silica gel (45 g), the elution proceeding as follows: 10% methanol / 0.5% ammonium hydroxide / chloroform (200 mL), nothing; (50 mL), unweighted impurity; (25 mL), nothing; (75 mL), unweighted mixed fraction; (200 mL), 0.010 g (8%) of 3- (2-chloro-pyridin-3-yl) -6-fluoro2 - [(3-pyrrolidin-1-ylmethyl-phenylamino) -methyl] -3H-quinazolin- 4-one as a pale yellow solid that had:¹H NMR δ 8.60 (d, J = 5 Hz, 1H), 7.80 (t, J = 5 Hz, 1H), 7.76 (d, J = 9 Hz, 1H), 7.71 (dd , J = 3.9 Hz, 1H), 7.58-7.50 (m, 2H), 7.35 (dd, J = 5.8 Hz, 1H), 7.14-7.08 (m, 2H), 6.80 (dd, J = 4.9 Hz, 1H), 4.44 (s, 1H), 3.90-3.65 (m, 4H), 2.79 (s broad, 4 H), 1.90 (s broad, 4H); APCI MS m / z = 464 (P +¹).

Example 9

6-Fluoro-3- (2-methyl-pyridin-3-yl) -2 - [(3-pyrrolidin-1-ylmethyl-phenylamino) -methyl] -3H-quinazolin-4-one

A mixture of 3- (2-methyl-pyridin-3-yl) -6-fluoro-3,4-dihydro-quinazolin-4-one-2-carboxaldehyde (0.158 g, 0.56 mmol), 3-pyrrolidin- 1-ylmethylaniline (0.050 g, 0.28 mmol), and acetic acid (0.16 mL, 2.8 mmol) in dichloroethane (15 mL) was cooled to 0 ° C and sodium triacetoxyborohydride (0.297 g, 1 , 4 mmol). The reaction was stirred for 1 hour at 0 ° C then was allowed to warm to room temperature and stirred for 16 hours. The reaction was stopped by the addition of saturated aqueous bicarbonate and stirring was continued for 30 minutes. The phases were separated and the aqueous layer was extracted with methylene chloride. The combined organic phase was washed with brine, dried over magnesium sulfate, and concentrated. The residue was subjected to flash chromatography on silica gel (35 g), elution proceeding as follows: 10% methanol / 0.5% ammonium hydroxide / chloroform (50 mL), nothing; (75 mL), unweighted impurity; (50 mL), 3- (2-methyl-pyridin-3-yl) -6-fluoro-3,4-dihydro-quinazolin-4-one-2-methanol unweighted; (50 mL), unweighted mixed fraction; (100 mL), 0.031 g (25%) of 3- (2-methyl-pyridin-3-yl) -6-fluoro2 - [(3-pyrrolidin-1-ylmethyl-phenylamino) -methyl] -3H-quinazolin- 4-one as a yellow foam that had: ¹H NMR δ 8.72 (d, J = 5 Hz, 1H), 7.92 (dd, J = 3.8 Hz, 1H), 7.82 (dd, J = 5.9 Hz, 1H), 7 , 58-7.52 (m, 2H), 7.39 (dd, J = 5.8 Hz, 1H), 7.08 (t, J = 8 Hz, 1H), 6.67-6.64 ( m, 2H), 6.37 (d, J = 8 Hz, 1H), 5.10 (s, 1H), 3.90 (dd, J = 4.5, 17 Hz, 1H), 3.70 (dd, J = 5.17 Hz, 1H), 3.56 (c AB, Av ^ = 21 Hz, J = 12.5 Hz, 2H), 2.54 (s, 4H), 2.36 (s, 3H) ; 1.78 (s, 4H), APCI MS m / z = 444 (P +¹).

ES 2 245 015 T3

Example 12

3- (2-chloro-phenyl) -6-fluoro-2 - [(3-pyrrolidin-1-ylmethyl-phenylamino) -methyl] -3H-quinazolin-4-one

A mixture of 3- (2-chloro-phenyl) -6-fluoro-3,4-dihydro-quinazolin-4-one-2-carboxaldehyde (0.50 g, 1.65 mmol), 3-pyrrolidin-1-ylmethylaniline ( 0.291 g, 1.65 mmol) and anhydrous sodium sulfate (2.3 g, 16.5 mmol) in dichloroethane (20 mL) was refluxed for 24 hours. The mixture was diluted with water and stirred for 20 min. The phases were separated and the organic phase was washed with saturated aqueous bicarbonate and brine, dried over magnesium sulfate, and concentrated to give 0.61 g of crude imine. The imine was dissolved in ethanol (20 mL) and 10% palladium on carbon (0.61 g) and formic acid (2.6 mL, 69.7 mmol) were added. The reaction was stirred at room temperature for 3 hours and then saturated aqueous sodium bicarbonate was added. The mixture was filtered through celite and the filtrate was concentrated to remove most of the ethanol. The milky aqueous residue was extracted with ethyl acetate. The organic phase was washed with water and brine, dried over magnesium sulfate, and concentrated. The residue was subjected to flash chromatography on silica gel (45 g), the elution proceeding as follows: 10% methanol / 0.5% ammonium hydroxide / chloroform (200 mL), nothing; (200 mL), 0.322 g (53%) 3- (2-chloro-phenyl) -6-fluoro-2 - [(3-pyrrolidin-1-ylmethyl-phenylamino) -methyl] 3H-quinazolin-4-one like a whitish foam that had: pf 105-110 ° C; Ή NMR δ 7.92 (dd, J = 3.8 Hz, 1H), 7.80 (dd, J = 5.9 Hz, 1H), 7.67-7.65 (m, 1H), 7, 59-7.50 (m, 3H), 7.40-7.38 (m, 1H), 7.08 (t, J = 8Hz, 1H), 6.696.65 (m, 2H), 6.40 (d, J = 8 Hz, 1H), 5.13 (s, 1H), 3.94 (dd, J = 5.17 Hz, 1H), 3.81 (dd, J = 4.5, 17 Hz , 1H), 3.59 (s broad, 2H); 2.58 (s broad, 4H), 1.80 (s broad, 4H). Analysis calculated for C₂₆H₂₄ClFN₄Oh₂O: C, 64.93; H, 5.45; N, 11.65. Found: C, 65.09; H, 5.04; N, 11.48.

Example 13

2- {[3- (2-chloro-phenyl) -6-fluoro-4-oxo-3,4-dihydro-quinazolin-2-ylmethyl] -amino} -nicotinonitrile

A mixture of 3- (2-chloro-phenyl) -6-fluoro-3,4-dihydro-quinazolin-4-one-2-carboxaldehyde (0.439 g, 1.45 mmol), 2-aminonicotinonitrile (0.150 g, 1.26 mmol) and anhydrous sodium sulfate (2.1 g, 14.5 mmol) in acetic acid (10 mL) was stirred at room temperature for 24 hours. The mixture was heated at this time at 80 ° C for 24 hours. The reaction was cooled to room temperature and sodium triacetoxyborohydride (1.3 g, 6.13 mmol) was added portionwise. The reaction was stirred for 16 hours and then stopped by carefully pouring it into a mixture of saturated aqueous sodium bicarbonate (150 mL) and ethyl acetate (20 mL). This stopping solution was stirred 30 min. The phases were separated and the organic layer was washed with brine, dried over magnesium sulfate, and concentrated. The residue was flash chromatographed on silica gel (50 g), elution proceeding as follows: 20% ethyl acetate / hexane (350 mL), nothing; 30% ethyl acetate / hexane (350 mL), unweighted 3- (2-chloro-phenyl) -6-fluoro-3,4-dihydroquinazolin-4-one-2-methanol; 40% ethyl acetate / hexane (400 mL), impure product. This partially purified product was flash chromatographed on silica gel (20 g), elution proceeding as follows: 5% ethyl acetate / hexane (1000 mL), nothing; 10% ethyl acetate / hexane (1,000 mL), nothing; 15% ethyl acetate / hexane (750 mL), nothing; (500 mL), unweighted impurity; 20% ethyl acetate / hexane (1,000 mL), nothing; (1,000 mL), unweighted impurity; 30% ethyl acetate / hexane (600 mL), 0.050 g (10%) of 2 - {[3- (2-chloro-phenyl) -6-fluoro-4-oxo-3,4-dihydro-quinazoline-2- ylmethyl] -amino} -nicotinonitrile as a light tan solid having: mp 145-150 ° C;¹H NMR δ 8.16 (d, J = 5 Hz, 1H), 7.93 (dd, J = 3.8 Hz, 1H), 7.84 (dd, J = 5.9 Hz, 1H), 7 , 71-7.63 (m, 2H), 7.56-7.51 (m, 3H), 7.40 (d, J = 7Hz, 1H), 6.85 (s, 1H), 6 .62 (dd, J = 5.7 Hz, 1H), 4.23 (c AB, Av_1-3 = 74 Hz, J = 19 Hz, 2H); APCI MS m / z = 406.2 (P +¹).

Example 14

3- (2-chloro-pyridin-3-yl) -6-fluoro-2 - [(2-fluoro-phenylamino) -methyl] -3H-quinazolin-4-one

A mixture of 3- (2-chloro-pyridin-3-yl) -6-fluoro-3,4-dihydro-quinazolin-4-one-2-carboxaldehyde (0.20 g, 0.66 mmol), 2- fluoroaniline (0.053 mL, 0.55 mmol) and anhydrous sodium sulfate (0.9 g, 6.6 mmol) in dichloroethane (10 mL) were refluxed for 18 hours. The reaction was concentrated and the residue was partitioned between ethyl acetate and water. The phases were separated and the organic phase was washed with saturated aqueous bicarbonate and brine, dried over magnesium sulfate, and concentrated to give 0.186 g of crude imine as a yellow foam. The imine was dissolved in ethanol (10 mL) and 10% palladium on carbon (0.186 g) and formic acid (0.90 mL, 23.5 mmol) were added. The reaction was stirred at room temperature for 1.5 hours. The acid was quenched by the addition of solid potassium carbonate and stirred 30 minutes. The mixture was filtered through celite and the bed was washed with ethanol and ethyl acetate. The filtrate was concentrated and the residue was taken up in ethyl acetate. This organic phase was washed with water and brine, dried over magnesium sulfate, and concentrated. The residue was flash chromatographed on silica gel (13 g), elution proceeding as follows: 10% ethyl acetate / hexane (350 mL), nothing; 20% ethyl acetate / hexane (50 mL), 0.012 g, recovered imine; (150 mL), nothing; (250 mL), 0.048 g (24%) 3- (2-chloro-pyridin-3-iI) -6-fluoro-2 - [(2-fluoro-phenylamino) -methyl] -3H-quinazolin-4-one as a white solid having: mp 180-182 ° C;¹H NMR δ 8.62 (d, J = 5 Hz, 1H), 7.95-7.91 (m, 2H), 7.80 (d, J = 8 Hz, 1H), 7.57 (dt, J = 3.8Hz, 1H), 7.50 (dd, J = 5.8Hz, 1H), 7.016.91 (m, 2H), 6.71 (dd, J = 7.12Hz, 1H), 6.53 (t, J = 9 Hz, 1H), 4.05 (c AB, Av₁-₃ = 19 Hz, J = 17 Hz, 2H); APCI MS m / z = 399 (P +¹).

ES 2 245 015 T3

Example 15

3- (2-chloro-phenyl) -6-fluoro-2 - [(2-fluoro-phenylamino) -methyl] -3H-quinazolin-4-one

A mixture of 3- (2-chloro-phenyl) -6-fluoro-3,4-dihydro-quinazolin-4-one-2-carboxaldehyde (0.10 g, 0.33 mmol), 2-fluoroaniline (0.064 mL, 0 , 66 mmol) and acetic acid (0.038 mL, 0.66 mmol) in methanol (10 mL) was stirred 1.5 hours at room temperature. Sodium cyanoborohydride (0.083 g, 1.32 mmol) was added and the reaction was stirred overnight. The reaction was diluted with water and concentrated to remove most of the methanol. The milky white liquid residue was treated with saturated aqueous bicarbonate and extracted with ethyl acetate. The organic layer was washed with aqueous bicarbonate and brine, dried over magnesium sulfate, and concentrated. The residue was flash chromatographed on silica gel (20 g) eluting the product with 5% and then 10% ethyl acetate / hexane. In this way 0.10 g (76%) of 3- (2-chloro-phenyl) -6-fluoro-2 - [(2-fluoro-phenylamino) -methyl] -3H-quinazolin-4one was isolated as a foam yellow that had: ¹H NMR δ 7.95 (dd, J = 3, 8.5 Hz, 1H), 7.85 (dd, J = 5.9 Hz, 1H), 7.69 (dd, J = 2, 7.5 Hz, 1H), 7.63-7.46 (m, 3H), 7.43-7.37 (m, 1H), 7.05-6.90 (m, 2H), 6.70-6 , 62 (m, 1H), 6.43 (dt, J = 1.9 Hz, 1H), 5.30 (s broad, 1H), 3.91 (c AB, Δν₁ -₃ = 27.5 Hz, J = 17 Hz, 2H). Analysis calculated for C₂₁H₁₄CIF₂N₃O-0.25 H₂O: C; 62.70; H, 3.63; N, 10.44. Found: C, 62.78; H, 3.51; N, 10.29.

Example 16

3- (2-chloro-phenyl) -6-fluoro-2 - [(6-methyl-pyridin-2-ylamino) -methyl] -3H-quinazolin-4-one

A mixture of 3- (2-chloro-phenyl) -6-fluoro-3,4-dihydro-quinazolin-4-one-2-carboxaldehyde (0.170 g, 0.56 mmol) and 2-amino-6-picoline ( 0.050 g, 0.46 mmol), in methanol (10 mL) refluxed overnight. The reaction was concentrated and the residue was flash chromatographed on silica gel (25 g). Elution proceeded as follows: 10% ethyl acetate / hexane (500 mL), nothing; 20% ethyl acetate / hexane (300 mL), nothing; (100 mL), 0.082 g of intermediate imine; (300 mL), 0.192 g of a mixture of starting material and imine. Pure imine (0.082 g, 0.21 mmol) was dissolved in ethanol (10 mL) and 10% palladium on carbon (0.082 g) and formic acid (0.357 mL, 9.45 mmol) were added. The reaction was stirred at room temperature for 24 hours. The reaction was carefully neutralized with sodium bicarbonate and then filtered through celite. The bed was washed well with ethyl acetate. The filtrate was concentrated to give 0.068 g (82%) of 3- (2-chloro-phenyl) -6-fluoro-2 - [(6-methyl-pyridin-2-ylamino) methyl] -3H-quinazolin-4- one as a colorless oil that had:¹H NMR δ 7.92 (dd, J = 3.8 Hz, 1H), 7.80 (dd, J = 5.9 Hz, 1H), 7.65-7.64 (m, 1H), 7, 53-7.48 (m, 4H), 7.43-7.40 (m, 1H), 7.37-7.31 (m, 1H), 6.47 (d, J = 7Hz, 1H ), 6.25 (d, J = 8 Hz, 1H), 4.18 (dd, J = 5.18 Hz, 1H), 4.00 (dd, J = 5.18 Hz, 1H), 2, 37 (s, 3H); APCI MS m / z = 395 (P +¹). Example 17

3- (2-chloro-phenyl) -2 - [(2-fluoro-phenylamino) -methyl] -3H-thieno [3,2-d] pyrimidin-4-one

A mixture of 3- (2-chloro-phenyl) -4-oxo-3,4-dihydro-thieno [3,2-d] pyrimidine-2-carboxaldehyde (0.435 g, 1.5 mmol), 2-fluoroaniline ( 0.121 mL, 1.25 mmol), and anhydrous sodium sulfate (2.1 g, 15 mmol) in glacial acetic acid (10 mL) was stirred overnight at room temperature. Sodium triacetoxyborohydride (0.795 g, 3.75 mmol) was added in one portion and the reaction was stirred for four hours. The reaction was carefully poured into saturated aqueous sodium bicarbonate and repeatedly extracted with methylene chloride. The combined organic phase was washed with water and brine, dried over sodium sulfate, and concentrated. The residue was flash chromatographed on silica gel eluting with hexane / ethyl acetate 2: 1. After elution of a 50 mL head, 15 mL fractions were collected. Fractions 3-12 were combined and concentrated to give 0.190 g (39%) of 3- (2-chlorophenyl) 2 - [(2-fluoro-phenylamino) -methyl] -3H-thieno [3,2-d] pyrimidin-4-one as a tan solid having: mp 175-176 ° C; ¹H NMR δ 7.84 (d, J = 5.5 Hz, 1H), 7.66 (dd, J = 1.5, 8 Hz, 1H), 7.53 (m symmetric, 2H), 7.42 (d, J = 5.5 Hz, 1H), 7.37 (dd, J = 2.7 Hz, 1H), 6.97 (ddd, J = 1.5, 8, 11.5 Hz, 1H) , 6.90 (t, J = 7.5 Hz, 1H), 6.63 (m symmetric, 1H); 6.40 (dt, J = 1.5, 9 Hz, 1H); 5.17 (s broad, 1H); 3.96 (dd, J = 5.5, 17 Hz, 1H), 3.87 (dd, J = 5.17 Hz, 1H). Analysis calculated for C₁₉H₁₃CIFN₃OS: C, 59.15; H, 3.40; N, 10.89. Found: C, 58.96; H; 3.41; N, 11.17.

Example 18

3- (2-Chloro-phenyl) -2 - [(3-pyrrolidin-1-ylmethyl-phenylamino) -methyl] -3H-thieno [3,2-d] pyrimidin-4-one hydrochloride

To a mixture of 3- (2-chloro-phenyl) -4-oxo-3,4-dihydro-thieno [3,2-d] pyrimidine-2-carbaldehyde (0.326 g, 1.13 mmol) and 3- ( Pyrrolidin-1-yl-methyl) -aniline (0.132 g, 0.75 mmol) in dichloroethane (10 mL) was added sodium triacetoxyborohydride (0.477 g, 2.25 mmol) portionwise. The reaction was stirred for 24 hours, then carefully poured into saturated aqueous sodium bicarbonate and repeatedly extracted with chloroform. The combined organic phase was washed with water and brine, dried over magnesium sulfate, and concentrated. The residue was flash chromatographed on silica gel (12 g, packed in chloroform). After eluting with chloroform (25 mL), elution was continued with 2-5% methanol / chloroform. The fractions containing the product (TLC on silica gel R_F = 0.20 (9: 1 chloroform / methanol with 1% triethylamine)] were combined and concentrated to give 0.166 g (53%) of the free base of the title product as a viscous oil had: ¹H NMR δ 7.84 (d, J = 5.5 Hz, 1H), 7.65 (m, 1H), 7.52 (m symmetric, 2H), 7.40 (d, J = 5 Hz, 1H ), 7.37 (m, 1H), 7.07 (t, J = 8Hz, 1H), 6.66 (d, J = 7.5Hz, 1H), 6.61 (s, 1H), 6.37 (dd, J = 2.8 Hz, 1H), 5.02 (t broad, J = 5 Hz, 1H), 3.94 (dd, J = 5.17 Hz, 1H), 3.84 (dd, J = 5.17 Hz, 1H), 3.51 (s, 2H), 2.48 (s broad, 4H), 1.75 (s broad, 4H). This oil was taken up in ether and treated with ethereal solution of

ES 2 245 015 T3 1N hydrochloric acid (0.75 mL, 0.75 mmol). The resulting thick suspension was concentrated and dried to give 0.115 g (31%) of 3- (2-chloro-phenyl) -2 - [(3-pyrrolidin-1-ylmethyl-phenylamino) -methyl] -3H- hydrochloride thieno [3,2-d] pyrimidin-4one as a tan solid having: mp 148-151 ° C.

Example 19

3- (2-chloro-phenyl) -6-fluoro-2- (2-fluoro-benzylamino) -3H-quinazolin-4-one

A suspension of 265 mg (0.87 mmol) of 3- (2-chloro-phenyl) -6-fluoro-2-thioxo-2,3-dihydro-1H-quinazolin-4-one in 2.0 mL of POCI3 It was treated with 300 mg of PCI5 and the mixture was refluxed for 2.5 hours. The solvent was evaporated, the residues were taken up in ethyl acetate, washed with water and brine, and then the solvent was evaporated to give

2- chloro-3- (2-chloro-phenyl) -6-fluoro-3H-quinazolin-4-one as a solid. This material was dissolved in 10 mL of absolute ethanol and 216 mg (1.73 mmol) of 2-fluorobenzylamine and the mixture was refluxed overnight. The reaction mixture was cooled and the solvent was evaporated. The residues were partitioned between ethyl acetate and dilute HCl and the aqueous phase was further extracted with ethyl acetate. The combined organic extracts were dried over brine and over magnesium sulfate (MgSO₄) and evaporated to give a gum which was crystallized from ether to give 157 mg (45%) of the desired product, mp 163-165 ° C.

Example 20

3- (2-chloro-phenyl) -6-fluoro-2 - [(pyridin-2-ylmethyl) -amino] -3H-quinazolin-4-one

A suspension of 306 mg (1.0 mmol) of 3- (2-chloro-phenyl) -6-fluoro-2-thioxo-2,3-dihydro-1H-quinazolin-4-one in 2.0 mL of POCI3 It was treated with 350 mg of PCI5 and the mixture was refluxed for 2.5 hours. The solvent was evaporated and the residues were taken up in ethyl acetate, washed with water and brine, and then the solvent was evaporated to give 2-chloro-3- (2-chloro-phenyl) -6-fluoro-3H-quinazolin -4-one as a solid. This material was dissolved in 15 mL of absolute ethanol and 238 mg (2.20 mmol) of 2-aminomethylpyridine and the mixture was refluxed overnight. The reaction mixture was cooled and the solvent was evaporated. The residue was dissolved in ethyl acetate and washed twice with water. The organic layer was dried with brine and with MgSO₄ and evaporated to give a gum which was crystallized from ether to give 190 mg (50%) of the desired product, mp 156-158 ° C.

Preparation 1

3- (2-chloro-pyridin-3-yl) -6-fluoro-3,4-dihydro-quinazolin-4-one-2-carboxaldehyde

A mixture of 3- (2-chloro-pyridin-3-yl) -6-fluoro-2-methyl-3,4-dihydro-quinazolin-4-one (3.5 g, 12.0 mmol) and dimethylformamide dimethyl Acetal (3.2 mL, 24 mmol) in dimethylformamide (12 mL) was refluxed for 24 hours. The reaction was cooled to room temperature and concentrated under reduced pressure to give an orange solid. The solid was triturated with ethanol and the yellow crystalline solid was collected and dried to give 3.61 g (87%) of 3- (2-chloro-pyridin-3-yl) -6-fluoro2- (2-dimethylamino- vinyl) -3,4-dihydro-quinazolin-4-one, having: mp 197-200 ° C; ¹H NMR δ 8.46 (dd, J = 1.5, 5 Hz, 1H), 7.85 (d, J = 12.2Hz, 1H), 7.70 (dd, J = 3.5 Hz , 1H), 7.68-7.65 (m, 1H), 7.45-7.38 (m, 2H), 7.31 (dt, J = 3, 8.5 Hz, 1H), 3, 87 (d, J = 12.2Hz, 1H), 2.80 (bs, 6h). Analysis calculated for C₁₇H₁₄CIFN₄O: C, 59.22; H; 4.09; N, 16.25. Found: C, 59.14; H, 3.96; N, 16.25.

To a mixture of sodium periodate (1.9 g, 8.7 mmol) in tetrahydrofuran (10 mL) and aqueous buffer of pH 7 (15 mL) was added 3- (2-chloro-pyridin-3-yl) -6-fluoro-2- (2-dimethylamino-vinyl) -3,4-dihydro-quinazolin-4-one (1.0 g, 2.90 mmol) all at once. The reaction was stirred for 1 hour. The precipitate was collected, washed well with water, and dried under a stream of nitrogen to give 0.863 g (92%) of 3- (2-chloro-pyridin-3-yl) -6-fluoro-3,4-dihydro -quinazoline-4one-2-carboxaldehyde as a yellow solid that was approximately a 25: 1 mixture of hydrate and free aldehyde. The product was characterized as follows: mp 160-164 ° C;¹H NMR (DMSO_d6) δ (hydrate) 8.51 (dd, J = 2.5 Hz, 1H), 8.08 (dd, J = 2.8 Hz, 1H), 7.87-7.78 (m, 3H) , 7.60 (dd, J = 5.8 Hz, 1H), 6.64 [c AB, Dn_M = 18 Hz, J = 7 Hz, 2H (OHs of the hydrate)], 5.27 [s broad, 1H (methine CH of the hydrate)]. The addition of D₂Or the NMR sample caused the multiplet to disappear at 6.64 ppm and the singlet to be further defined at 5.27 ppm. The presence of the free aldehyde was demonstrated by a singlet less than 9.51 ppm.

Preparation 2

3- (2-chloro-phenyl) -6-fluoro-3,4-dihydro-quinazolin-4-one-2-carboxaldehyde

A mixture of 3- (2-chloro-phenyl) -6-fluoro-2-methyl-3,4-dihydro-quinazolin-4-one (1.0 g, 3.46 mmol) and dimethylformamide dimethyl acetal (0, 92 mL, 6.92 mmol) in dimethylformamide (4 mL) was heated at 140 ° C for 24 hours. The reaction was cooled to room temperature and concentrated under reduced pressure. The dark residue was triturated with methanol and the strong yellow crystalline solid that formed was collected and dried to give 1.075 g (90%) of 3- (2-chloro-phenyl) -6-fluoro-2- (2-dimethylamino-vinyl ) -3,4-dihydro-quinazolin-4-one having: mp 210-211 ° C; ¹H NMR δ 7.86 (d, J = 12.3 Hz, 1H), 7.79 (dd, J = 3, 8.5 Hz, 1H), 7.61-7.54 (m, 1H), 7.51-7.29 (m, 5H), 4.06 (d, J = 12.3Hz, 1H), 2.80 (bs, 6H). Analysis calculated for C₁₈H₁₅CIFN₃O: C, 62.89; H, 4.40; N, 12.22. Found: C, 62.75; H, 4.28; N, 12.29.

ES 2 245 015 T3

To a well stirred mixture of sodium periodate (2.24 g, 10.47 mmol) in a pH 7 buffer (10 mL) and tetrahydrofuran (10 mL) was added 3- (2-chloro-phenyl) -6 -fluoro-2- (2-dimethylamino-vinyl) -3,4-dihydro-quinazolin-4-one (0.90 g, 2.62 mmol) all at once. The mixture was slightly warm to the touch and stirred 1 hr at room temperature. The reaction was filtered through celite and the bed was rinsed with ethyl acetate. The phases were separated from the filtrate and the aqueous layer was extracted with ethyl acetate. The combined organic phase was washed with brine, dried over magnesium sulfate, and concentrated to give 0.802 g (97%) of 3- (2-chloro-phenyl) -6-fluoro-3,4-dihydro-quinazoline- 4-one-2-carboxaldehyde as a 1: 2 mixture of free aldehyde and hydrate having: 'II NMR δ 9.52 (s) [CHO], 8.20-7.45 (m, 7H), 6 , 75 (d, J = 7.4 Hz) [OH of the hydrate] and 6.49 (d, J = 8 Hz) [OH of the hydrate], 5.14 (t, J = 7.5 Hz) [methine CH of the hydrate].

Preparation 3

6-Fluoro-3,4-dihydro-3- (2-methyl-pyridin-3-yl) -quin, azolin-4-on, a-2-carboxaldehyde

A mixture of 6-fluoro-3,4-dihydro-2-methyl-3- (2-methyl-pyridin-3-yl) -quinazolin-4-one (1.02 g, 3.79 mmol) and dimethyl dimethylformamide Acetal (1.01 mL, 7.58 mmol) in dimethylformamide (5 mL) was heated at 140 ° C for 24 hours. More dimethylformamide dimethyl acetal (2 mL) was added and the reaction heated at 140 ° C for 16 hours. The reaction was cooled to room temperature and concentrated under reduced pressure. The dark residue was triturated with ether and the dark purple crystalline solid that formed was collected and dried to give 0.69 g (56%) of 6-fluoro-2- (2-dimethylaminovinyl) -3,4-dihydro- 3- (2-methyl-pyridin-3-yl) -quinazolin-4-one having:¹H NMR δ 8.61 (dd, J = 1.5, 5 Hz, 1H), 7.87 (d, J = 12.3 Hz, 1H), 7.78 (dd, J = 3, 8.5 Hz, 1H), 7.50-7.45 (m, 2H), 7.39-7.29 (m, 2H), 3.97 (d, J = 12.3 Hz, 1H), 2.86 (s broad, 6H), 2.35 (s, 3H).

To a well stirred mixture of sodium periodate (1.76 g, 8.26 mmol) in a pH 7 buffer (10 mL) and tetrahydrofuran (15 mL) was added 6-fluoro-2- (2-dimethylamino- vinyl) -3,4-dihydro-3- (2-methyl-pyridin-3-yl) -quinazolin-4one (0.69 g, 2.12 mmol) all at once. The mixture was slightly warm to the touch and stirred for 1 hour at room temperature. The reaction was concentrated under reduced pressure and the residue was treated with a 1: 2 mixture of water and chloroform. The tan solid was collected, washed well with water and chloroform, and dried under a stream of nitrogen to give 0.50 g (80%) of 6-fluoro-3,4-dihydro-3- (2-methyl -pyridin-3-yl) -quinazolin-4-one-2-carboxaldehyde as a 1: 1 mixture of free aldehyde and hydrate having: mp 175-177 ° C; ¹H NMR δ 9.46 (s, 0.5H) [CHO], 8.53 (s broad, 1H), 8.10-8.07 (s, 0.5H), 7.94-7, 70 (m, 3.5H), 7.39-7.32 (m, 1H), 6.59 (s broad, 0.5H) [OH of hydrate], 6.47 (s broad, 0, 5H) [OH of the hydrate], 5.17 (s, 0.5H) [methine CH of the hydrate], 2.20 (s, 3H); ApCi MS m / z = 284.1 (P +¹).

Preparation 4

3- (2-chloro-phenyl) -4-oxo-3,4-dihydro-thieno [3,2-d] pyrimidine-2-carbaldehyde

A mixture of 3- (2-chloro-phenyl) -2-methyl-4-oxo-3,4-dihydro-thieno [3,2-d] pyrimidine (9.4 g, 34.06 mmol) and dimethylformamide dimethyl Acetal (9.0, 68.12 mmol) in dimethylformamide (70 mL) was heated at 140 ° C for 24 hours. The reaction was cooled to room temperature and concentrated under reduced pressure (50 ° C thermostatic bath). The residue was suspended in methanol and concentrated under reduced pressure. This methanol suspension / concentration process was repeated twice. The residue was triturated with methanol and the light yellow crystalline solid that formed was collected and dried to give 10.1 g (85%) of 3- (2-chloro-phenyl) -2- (dimethylamino-vinyl) -4 -oxo-3,4-dihydro-thieno [3,2-d] pyrimidine having: ¹H NMR δ 7.83 (d, J = 12.5 Hz, 1H), 7.68 (d, J = 5 Hz, 1H), 7.57 (m, 1H), 7.41 (m, 2H) , 7.31 (m, 1H), 7.14 (d, J = 5Hz, 1H), 4.06 (d, J = 12.5Hz, 1H), 2.77 (s broad, 6H) .

To a well stirred mixture of sodium periodate (9.7 g, 45.3 mmol) in a pH 7 buffer (115 mL) and tetrahydrofuran (115 mL) was added 3- (2-chloro-phenyl) -2 - (dimethylamino-vinyl) -4-oxo-3,4-dihydro-thieno [3,2-d] pyrimidine (5.0 g, 15.1 mmol) all at once. The mixture was slightly warm to the touch and stirred for 1 hour at room temperature. The reaction was filtered through celite and the bed was washed with ethyl acetate. The phases were separated from the filtrate and the aqueous layer was extracted with ethyl acetate. The combined organic phase was washed with brine, dried over sodium sulfate, and concentrated to give a yellow oil. The oil was flash chromatographed on silica gel (70 g), elution proceeding as follows: 50% methylene chloride / hexane (500 mL), unweighted headless; 50% ethyl acetate / hexane (1,000 mL), 4.6 g (100%) of 3- (2-chloro-phenyl) -4-oxo-3,4-dihydro-thieno [3,2-d] pyrimidine-2-carbaldehyde as a foam that was a 2: 1 mixture of free aldehyde and hydrate and had:¹H NMR δ 9.56 (s) [CHO], 7.94 and 7.87 (pair of d, J = 5.2, 5.4 Hz, respectively, 1H), 7.60-7.30 (m , 5H), 5.30 (dd, J = 7, 9.5 Hz) [methine CH of the hydrate), 4.76 (d, J = 9.5 Hz) [OH of the hydrate], 3.79 ( d, J = 7 Hz) [OH of the hydrate]. Note that the doublets at 4.76 and 3.79 ppm are poorly defined and the dd at 5.30 ppm became a singlet when deuterium oxide was added to the NMR sample.

Preparation 5

3- (2-chloro-phenyl) -3,4-dihydro-quinazolin-4-one-2-carboxaldehyde (A) A mixture of 3- (2-chloro-phenyl) -2-methyl-3,4-dihydro -quinazolin-4-one (1.0 g, 3.69 mmol) and dimethylformamide dimethyl acetal (0.64 mL, 4.8 mmol) in dimethylformamide (4 mL) was heated at 140 ° C for 24 hours. More dimethylformamide dimethyl acetal (0.32 mL, 2.4 mmol) was added and heating continued for a further 24 hours. The reaction

ES 2 245 015 T3 was cooled to room temperature and concentrated under reduced pressure (thermostatic bath at 60 ° C). The dark residue was triturated with methanol and the pale orange solid that formed was collected and dried to give 0.75 g (62%) of 3- (2-chlorophenyl) -2- (2-dimethylamino-vinyl) -3 , 4-dihydro-quinazolin-4-one having: mp 228-230 ° C;¹H NMR δ 8.19 (d, J = 8 Hz, 1H), 7.91 (d, J = 12.3 Hz, 1H), 7.71-7.18 (m, 7H), 4.10 (d, J = 12.3 Hz, 1H); 2.81 (s broad, 6H). Analysis calculated for C₁₈H₁₈CIN₃O: C, 66.36; H, 4.95; N, 12.90. Found: C, 66.20; H, 5.03; N, 12.79.

(B) To a well stirred mixture of sodium periodate (1.38 g, 6.45 mmol) in water (6 mL) and tetrahydrofuran (6 mL) was added 3- (2-chloro-phenyl) -2- (2-dimethylamino-vinyl) -3,4-dihydro-quinazolin-4-one (0.70 g, 2.15 mmol) all at once. The mixture was slightly warm to the touch and stirred for 1 hour at room temperature. The reaction was filtered through celite and the bed was rinsed with ether. The filtrate was made basic by the addition of saturated aqueous sodium bicarbonate and the phases were separated. The aqueous layer was extracted with ethyl acetate. The combined organic phase was washed with brine, dried over potassium carbonate, and concentrated. The residue was flash chromatographed on silica gel (1 x 3 inches (2.5 x 7.6 cm)), elution proceeding as follows: 50% methylene chloride / hexane (400 mL), nothing; (100 mL) and 50% ethyl acetate / hexane (150 mL), 0.51 g (82%) of 3- (2-chloro-phenyl) -3,4-dihydroquinazolin-4-one-2-carboxaldehyde as an approximately 1: 1 mixture of free aldehyde and hydrate containing approximately 20% 3- (2-chlorophenyl) -3,4-dihydro-quinazolin-4-one-2-acetaldehyde (by simple hydrolysis of enamine; It exists exclusively in its enolized form to take advantage of an internal hydrogen bond with the N1 of quinazolin-4-one). The characteristics that identify each of the components of this mixture can be observed in the NMR spectrum as defined in the following table:

TABLE

Desplazamie nto NMR key component of Preparation 5 (ppm)

<td>2-carboxaldehyde</td><td>2-carboxaldehyde hydrate</td><td>2-Acetaldehyde enolized</td>

<td>9.60 (s) [CHO]</td><td>5.30 (m) [methine CH] 4.94 (d, J = 9.3 Hz) [OH] 3.90 (d, J = 7.2 Hz) [OH]</td><td>8.71 (d, J = 3.7 Hz) and 4.40 (d, J = 3.7 Hz) [enol protons, cis-coupled]</td>

Consistent with Preparations 1-4, running Preparation 5 in pH 7 buffered water will suppress the formation of the enolyzed 2-acetaldehyde by-product.

Preparation 6

3- (2-chloro-phenyl) -6-fluoro-2-thioxo-2,3-dihydro-1H-quinazolin-4-one

A solution of 1.04 g (6.71 mmol) of 5-fluoroanthranilic acid and 1.13 g (6.71 mmol) of 2-chlorophenylisothiocyanate in 10 ml of glacial acetic acid was refluxed for 2.5 hours. The reaction mixture was cooled and the acetic acid was evaporated to give a yellow solid residue, which was taken up in ethyl acetate and ether. The solid was filtered and washed with ether and air dried to give 1.48 g (72%) of the desired product, mp 295-297 ° C.

Separation of atropoisomers by HPLC Example 21

The HPLC separation of 3- (2-chloro-phenyl) -6-fluoro-2 - [(3-pyrrolidin-1-ylmethyl-phenylamino) -methyl) -3H-quinazolin-4-one atropoisomers is described below continuation.

<td>Column</td><td>Chiralpak AD</td>

<td>Mobile Phase</td><td>85/15 hexane / isopropyl alcohol with 0.1% diethylamine</td>

<td>Flow</td><td>1 ml / min</td>

<td>Detection</td><td>UV (250 nM)</td>

<td>Retention time (first atropoisomer)</td><td>13,635 min</td>

<td>Retention time (second atropoisomer)</td><td>16,509 min</td>

ES 2 245 015 T3

Example 22

The HPLC separation of the atropoisomers of 2 - ([3- (2-chloro-pyridin-3-yl) -6-fluoro-4-oxo-3,4-dihydro-quinazolin-2-ylmethyl] -amino} - benzonitrile is described below.

<td>Column</td><td>Chiralpak AD</td>

<td>Mobile Phase</td><td>70/30 hexane / isopropyl alcohol with 0.1% diethylamine</td>

<td>Flow</td><td>1 ml / min</td>

<td>Detection</td><td>UV (335 nM)</td>

<td>Retention time (first atropoisomer)</td><td>8,522 min</td>

<td>Retention time (second atropoisomer)</td><td>14,101 min</td>