8−quinolinxanthine and 8−isoquinolinxanthine derivatives as pde 5 inhibitors

The present invention relates to a compound derived from 8-quinolinoxanthine and 8-isoquinolinoxanthine, its use, a pharmaceutical composition containing the compound, and a method for producing the compound.

From WO 99/62905, 8-phenylxanthine derivatives of the formula indicated in the cited document are known having strong and selective inhibitory properties against cyclic guanosine 3 ', 5'-monophosphate phosphodiesterases. These derivatives have a phenyl group in the molecule and an associated -SO2NR group associated with it⁴R⁵.

From US 4,599,338 xanthine derivatives containing an 8- [3- (4-aminocarbonylpiperazine or -piperidine group) are known.

The subject of the invention is a new compound, 8-quinolinoxanthine and 8-isoquinolinoxanthine derivative, of formula I

<img file="PL212407B1_D0001.tif" />

in free form or in pharmaceutically acceptable salt form, where:

₁

R¹ is a hydrogen atom or a C1-C4-alkyl group;

₂

R² represents C1-C8-alkyl, hydroxy-C1-C8-alkyl, C1-C4-alkylcarbonyloxy-C1-C8-alkyl, C2-C4-alkenyl, C3-C6-cycloalkyl-C1-C4-alkyl, tetrahydrofuryl- ( C1-C4) -alkyl, methylcyclopropylmethyl or a phenyl-C1-C4-alkyl group in which the phenyl ring is optionally substituted with one or two substituents selected from: C1-C4-alkoxy, amino, C1-C4-alkylcarbonylamino, C1-C4-alkylsulfonylamino, di (C1-C4-alkyl) -sulfonylamino, halogen atom, di (C1-C4-alkyl) aminosulfonylamino group, or the phenyl ring can be fused to a 5-membered heterocyclic ring having two oxygen atoms,

R³ is a hydrogen atom or a C1-C4-alkyl group;

R⁴ is a hydrogen atom or a C1-C4-alkyl group;

₃

R³ is selected from the groups represented by the formulas II (quinolinyl group), III (isoquinolinyl group) or IIIA (oxohydroisoquinolinyl group):

<img file="PL212407B1_D0002.tif" />

PL 212 407 B1

R⁸ represents a hydrogen atom, a halogen atom, a cyano group, C1-C4-alkyl, di (C1-C4-alkyl) amino, morpholinyl, piperidyl, piperazinyl or a phenyl group substituted with one or two (C1-C4) -alkoxy groups

R⁹ is a hydrogen atom or a C1-C4-alkyl group

R¹⁰ is a hydrogen atom or a halogen atom

R¹¹ represents a hydrogen atom, a halogen atom, a C1-C4-alkoxy, hydroxyl, cyano or C2-C4-alkynyl group

R¹² is a hydrogen atom, a halogen atom, a C1-C4-alkoxy or a hydroxyl group

R¹¹ and r¹² together with the carbon atoms to which they are attached represent a 5-membered heterocyclic group having two ring oxygen atoms;

R¹³ is a hydrogen atom or a halogen atom

R³ represents a C1-C4-alkyl group

Among the compounds of formula I, preferred are those in which R² is as defined above, wherein halogen is only chlorine or bromine.

Preferably the compound of formula I is a compound wherein R⁵ represents an isoquinolinyl group of the formula III, where R⁸ represents a hydrogen atom, a C1-C4-alkyl group, a halogen atom, a cyano group; R⁹ is a hydrogen atom or a C1-C4-alkyl group; R¹⁰ is a hydrogen atom or a halogen atom; R¹¹ represents a hydrogen atom, a halogen atom, a hydroxyl group; C1-C4-alkoxy, C2-C4-alkynyl, cyano, R¹² is a hydrogen atom, a halogen atom, a hydroxyl group or a C1-C4-alkoxy group. R¹¹ and r¹² together with the carbon atoms to which they are attached represent a 5-membered heterocyclic group having two ring oxygen atoms; R¹³ is a hydrogen atom or a halogen atom.

It is also preferred that the compound of formula is a compound in which R¹ is -CH3, R.² is (CH3) 2CHCH2-, R³ and r⁴ are H, R.⁸ is -CH3, R.⁹ and r¹⁰ are H, R.¹¹ and r¹², are -OCH3

It is also preferred that a compound of formula I is a compound in which R¹ is -CH3, R.² is (CH3) 2CHCH2-, R³, R⁴, R⁸, R⁹ and r¹⁰ are H, R.¹¹ and r¹² are -OCH3.

It is also preferred that a compound of formula I is a compound in which R¹ is -CH3, R.² is (CH3) 3CCH2-, R³, R⁴, R⁸, R⁹ and r¹⁰ are H, R.¹¹ and r¹² are -OCH3.

It is also preferred that a compound of formula I is a compound in which R¹ is -CH3, R.² is (CH3) 2CHCH2-, R³, R⁴, R⁹ and r¹⁰ are H, R.⁸ is Cl, R.¹¹ and r¹² are -OCH3.

It is also preferred that a compound of formula I is a compound in which R¹ is -CH3, R.² is (CH3) 2CHCH2-, R³, R⁴, R⁸, R⁹ and r¹⁰ are H, R.¹¹ is -OCH3, R.¹² stands for H.

It is also preferred that a compound of formula I is a compound in which R¹ is -CH3, R.² is cyclopropylmethyl, R³, R⁴, R⁸, R⁹, R¹⁰ and r¹² are H, R.¹¹ stands for -OCH₃.

It is also preferred that a compound of formula I is a compound in which R¹ is -CH3, R.² means (CH₃)₂CHCH₂-, R³, R⁴, R⁸, R⁹, R¹⁰ and r¹² are H, R.¹¹ means CH C-.

It is also preferred that a compound of formula I is a compound in which R¹ is -CH3, R.² is a 4- (N-dimethylaminosulfonylamino) benzyl group, R³, R⁴, R⁸, R⁹ and r¹⁰ are H, R.¹¹ and r¹² are -OCH3.

It is also preferred that a compound of formula I is a compound in which R¹ is -CH3, R.² is HOCH2CH (CH3) CH2-, R³, R⁴, R⁸, R⁹ and r¹⁰ are H, R.¹¹ and r¹² are -OCH3.

It is also preferred that a compound of formula I is a compound in which R¹ is -CH3, R.² is 1-methylcyclopropylmethyl, R³, R⁴, R⁸, R⁹ and r¹⁰ are H, R.¹¹ and r¹² are -OCH₃.

The 8-quinolinoxanthine and 8-isoquinolinoxanthine derivative of formula I finds use as a medicament.

The invention also relates to a pharmaceutical composition comprising an active ingredient and a pharmaceutically acceptable solvent or carrier, wherein the active ingredient is a compound of formula I as defined above.

The invention relates to the use of a compound of formula I for the manufacture of a medicament for the treatment of conditions associated with pulmonary hypertension and sexual dysfunction, especially male sexual dysfunction, especially male erectile dysfunction.

The invention also relates to a process for the preparation of a compound of formula I in free form or in the form of a pharmaceutically acceptable salt by first dehydrating a compound of formula IV

PL 212 407 B1

<img file="PL212407B1_D0003.tif" />

where r¹, R², R⁴ and r⁵ are as defined for a compound of Formula I.

(b) where a compound of formula I is prepared in free form or in the form of a pharmaceutically acceptable salt, wherein R³ represents a C1-C4-alkyl group, reacting a compound of formula I in free form or in the form of a pharmaceutically acceptable salt with a suitable alkylating agent; or (c) where a compound of formula I is prepared in free form or in the form of a pharmaceutically acceptable salt, wherein R² represents a phenyl-C1-C4-alkyl group substituted in the phenyl ring with a C1-C4-alkylsulfonylamino group or a di (C1-C4-alkyl) aminosulfonylamino group, the compound of formula I is reacted in free form or in the form of a pharmaceutically acceptable salt, respectively - with a halide alkylsulfonate or dialkylaminosulfone halide; or (d) where a compound of formula I is prepared in free form or in the form of a pharmaceutically acceptable salt, wherein R² represents a hydroxyl-C1-C8-alkyl group, a hydration reaction of a compound of formula I is carried out in which R² is a C1-C8-alkenyl group; or (e) where a compound of formula I is prepared in free form or in the form of a pharmaceutically acceptable salt, wherein R² represents a C1-C4-alkylcarbonyloxy-C1-C8-alkyl group, an esterification reaction of a compound of formula I is carried out in which R² represents a C1-C8-alkyl group substituted with a hydroxyl group; or (f) where a compound of formula I is prepared in free form or in the form of a pharmaceutically acceptable salt, wherein R² represents a phenyl-C1-C4-alkyl group substituted in the phenyl ring with an amino group, the hydrolysis reaction of a compound of formula I is carried out in which R² represents a phenyl-C1-C4-alkyl group substituted on the phenyl ring with an acylamino group; or (g) where a compound of formula I is prepared in free form or in the form of a pharmaceutically acceptable salt, wherein R⁵ represents a quinolinyl or isoquinolinyl group substituted with a hydroxyl group, a dealkylation reaction of a compound of formula I is carried out in which R⁵ is respectively - a quinolinyl or isoquinolinyl group, substituted with an alkoxy group; or (h) where a compound of formula I is prepared in free form or in the form of a pharmaceutically acceptable salt, wherein R⁵ represents a quinolinyl or an isoquinolinyl group substituted with a halogen atom, a halogenation reaction of a compound of formula I is carried out in which R⁵ is, respectively, a quinolinyl or isoquinolinyl group having an unsubstituted ring carbon atom suitable for halogenation; or (i) where a compound of formula I is prepared in free form or in the form of a pharmaceutically acceptable salt, wherein R² represents a cyclopropyl group substituted with a C1-C4-alkyl group, a compound of formula I, wherein R² represents an alkenyl group undergoes a Simmons-Smith cyclopropanation reaction;

the compounds of formula I obtained are then isolated in free form or in the form of a pharmaceutically acceptable salt.

The invention also relates to a compound of formula IV

<img file="PL212407B1_D0004.tif" />

PL 212 407 B1 wherein R¹, R², R⁴ and r⁵ are as in the compound of formula I. The compound of formula IV is the starting compound for the preparation of the compound of formula I.

"Alkyl" means a straight chain or branched alkyl group which can be, for example, a C1-C10-alkyl such as: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, straight or branched pentyl, straight or branched hexyl, straight or branched heptyl, straight or branched octyl, straight or branched nonyl, straight or branched decyl.

"Alkoxy" means a straight or branched alkoxy group which can be, for example, C1-C10-alkoxy such as: methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, straight or branched pentoxy, straight or branched hexyloxy, straight or branched heptyloxy, straight or branched octyloxy, straight or branched nonyloxy, straight or branched decyloxy.

"Alkenyl" means a straight chain or branched alkenyl group which can be, for example, C2 to C10-alkenyl such as vinyl, 1-propenyl, 2-propenyl, 1-butenyl, isobutenyl or straight or branched pentenyl, hexenyl, heptenyl, octenyl , nonenyl or decenyl.

"Cycloalkylalkyl" means an alkyl group, for example C1 to C10-alkyl such as one of the C1-C10 alkyl groups mentioned above, substituted with C3 to C8-cycloalkyl such as: cyclopropyl, methylcyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methylcyclohexyl, dimethylcyclohexyl or cyclooctyl.

"Heterocycloalkyl" means an alkyl group, for example a C1 to C10-alkyl, such as one of the C1-C10 alkyl groups mentioned above, substituted with a 5-membered heterocyclic group having two ring oxygen heteroatoms.

"Acyl" means an alkylcarbonyl group, for example C1-C10-alkylcarbonyl where the C1-C10-alkyl may be one of the C1-C10-alkyl groups mentioned above, optionally substituted with one or more halogen atoms; cycloalkylcarbonyl, e.g. C3-C8-cycloalkylcarbonyl, where C3-C8-cycloalkyl may be e.g. cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl; A 5- or 6-membered heterocyclylcarbonyl group having one or two heteroatoms selected from: nitrogen, oxygen and ring sulfur, such as furylcarbonyl or pyridylcarbonyl; arylcarbonyl, e.g. C6-C10-arylcarbonyl such as benzoyl; or an aralkylcarbonyl group, for example C6 to C10-aryl-C1-C4-alkylcarbonyl such as benzylcarbonyl or phenylethylcarbonyl.

"Alkynyl means a straight or branched alkynyl group, for example C2-C4-alkynyl such as: ethynyl, propargyl, 2-butynyl.

"Aryl" means a monovalent carbocyclic aromatic group, for example a C6-C10-aryl such as: phenyl, phenyl substituted with one or more, e.g. one, two, or three C1-C4-alkyl groups, or naphthyl.

"Heteroaryl" means a monovalent aromatic heterocycle having 5 ring atoms, two of which are oxygen.

"Halogen" means fluoro, chloro, bromo or iodo.

Compounds of formula I may exist in the form of pharmaceutically acceptable salts. The pharmaceutically acceptable salts of the compounds of formula I include the salts of inorganic acids, for example, hydrohalic acids such as hydrofluoric acid, hydrochloric acid, hydrobromic acid or hydroiodic acid, nitric acid, sulfuric acid, phosphoric acid; organic acids, for example aliphatic monocarboxylic acids such as formic acid, acetic acid, trifluoroacetic acid, propionic acid and butyric acid, aliphatic hydroxy acids such as lactic acid, citric acid, tartaric acid or malic acid, dicarboxylic acids such as maleic acid or succinic acid , aromatic carboxylic acids such as benzoic acid, p-chlorobenzoic acid, diphenylacetic acid, or triphenylacetic acid, aromatic hydroxy acids such as o-hydroxybenzoic acid, p-hydroxybenzoic acid, 1-hydroxynaphthalene-2-carboxylic acid or 3-hydroxynaphthalene-2-carboxylic acid, and sulfonic acids such as methanesulfonic acid or benzenesulfonic acid. The pharmaceutically acceptable salts of compounds of formula I with bases wherein R³ is hydrogen, especially the salts of alkali metals or alkaline earth metals such as sodium, potassium, magnesium or calcium, salts with ammonia or pharmaceutically acceptable organic amines such as ethanolamines or benzylamines. These salts can be prepared from the free compounds of formula I or other salts of compounds of formula I according to known salt formation methods.

The individual steps for preparing a compound of the formula I can be carried out as follows:

PL 212 407 B1

Process (a) may be carried out by heating or by reaction with inorganic or organic bases. This can be done in an organic or aqueous solvent or a water / organic solvent mixture. The reaction with the base can be carried out at ambient temperature or, more preferably, at elevated temperature. Preferably, the reaction is carried out by treatment with aqueous solutions of alkali metal hydroxides in an alcoholic solvent at elevated temperature, for example as described below in the Examples. The compound of formula IV is preferably a compound where R⁵ is a group of formula II or III. Compounds of formula IV can be obtained by reacting a compound of formula

<img file="PL212407B1_D0005.tif" />

where R and R are defined above, with a compound of formula

HOOC-CH-R⁵

R⁴ or an amide-forming derivative thereof, where R⁴ and r⁵ are defined above. The reaction can be carried out by treating a carboxylic acid of formula VI with a peptide coupling agent to form an active ester or mixed anhydride in situ, followed by reacting with a compound of formula V in an organic, e.g. dipolar aprotic, or mixed organohydrocarbon (e.g. chlorohydrocarbon). solvent. The second step of the reaction may be carried out at reduced, room or elevated temperature, preferably at room temperature. Preferably, the acid of formula VI is reacted with the carbodiimide derivative in the presence of hydroxybenzotriazole and optionally a base, or is reacted with a benzotriazolyl- (trisdialkylamino) oxyphosphonium salt. The resulting intermediate is preferably treated with a compound of formula V in a dipolar aprotic solvent or a mixed (hydrogen chloride-water) solvent at ambient temperature. The procedures may be as described below in the examples.

Compounds of formula V can be obtained by reducing a compound of formula

<img file="PL212407B1_D0006.tif" />

where r¹ and r² are defined above.

The reduction can be carried out using known methods, for example by treating a compound of the Formula VII with a reducing agent in an organic or aqueous solvent. The reaction can be performed at room or, more preferably, at elevated temperature. Preferred reducing agents are alkali metal dithionates in an aqueous medium or hydrogen in the presence of a noble metal catalyst. The reaction with sodium dithionite in aqueous solution at 80-90 ° C is particularly preferred.

Compounds of formula VII can be obtained by nitrosating compounds of formula

PL 212 407 B1

<img file="PL212407B1_D0007.tif" />

where r¹ and r² are defined above, for example by means of an organic or inorganic nitrosating agent in an organic or inorganic or mixed aqueous-organic solvent. The nitrosation can be carried out according to known methods at reduced temperature, at ambient temperature or at elevated temperature, preferably with an alkali metal nitrite e.g. sodium nitrite in the presence of an acid e.g. acetic acid at reduced temperature or at ambient temperature, preferably in a mixed hydroalcoholic solution, e.g. water-ethanol.

Compounds of formula VIII can be obtained by reacting compounds of formula

R²—N-CO — N-CO — CH_?CN IX

Η H. ² where r² as defined above, with an inorganic or organic base in which cyclization takes place with the proviso that, if R¹ is a C1-C4-alkyl group by reaction with an alkylating agent. The cyclization is carried out in a manner generally known in the art, preferably in an aqueous, organic or mixed aqueous-organic solvent, at ambient temperature or more preferably at elevated temperature. The base used is an alkali metal hydroxide, e.g. sodium hydroxide, which is reacted in a mixed aqueous / alcoholic solvent, preferably at elevated temperatures, e.g. 80-90 ° C. The optional alkylation is carried out according to generally known methods, e.g. in the presence of an organic or inorganic base, e.g. in an aqueous, organic or mixed aqueous-organic solvent. The alkylation may be conducted below ambient temperature or more preferably at or above ambient temperature. Preferred alkylating agents are alkyl iodides or, especially, dialkyl sulfates. The preferred bases are alkali metal hydroxides in a water-alcohol mixture, especially a water-ethanol mixture.

Compounds of formula IX can be obtained by reacting a compound of formula

R — N — CO — NH₂ X

H. ² with cyanoacetic acid or with an amide-forming compound such as an acid ester or chloride, preferably an acid or an ethyl ester thereof. The reaction is carried out according to generally known methods in organic solvents, preferably in anhydrides such as acetic anhydride, at ambient temperature or most preferably at an elevated temperature, e.g. 65-70 ° C.

₂

Compounds of formula X can be obtained according to known methods, e.g. from the isocyanate R.²NCO by reaction with gaseous or aqueous ammonia or with the R amine²NH2 by reaction with metal cyanate as further described in the examples.

Compounds of formula VIII, wherein R¹ represents a C1-C4-alkyl group and R² as defined above, can be obtained by hydrogenating a compound of formula

<img file="PL212407B1_D0008.tif" />

PL 212 407 B1 where R.¹ represents a C1-C4-alkyl group, R² as defined above, Ar is a phenyl group optionally substituted with one or more C1-C4-alkoxy groups, preferably with a methoxy group. The hydrogenation may be carried out according to known methods, e.g. by using hydrogen gas or a hydrogen source in the presence of a catalyst, e.g. a platinum or preferably a palladium catalyst, at ambient temperature or at an elevated temperature. The reaction is carried out in an organic solvent. Preferably the hydrogenation is carried out using palladium black in formic acid as a catalyst as further described in the examples.

Compounds of formula XI were obtained by reacting a compound of formula

<img file="PL212407B1_D0009.tif" />

where r¹ and r² are as previously defined for a compound of formula XI with a compound of formula ArCH2NH2, where Ar is as defined above. The reaction may be carried out in a known manner, e.g. in an organic solvent, preferably an alcohol such as n-butanol at ambient or elevated temperature, or analogously as described further in the Examples.

The compound of formula XII can be obtained by reacting the compound

<img file="PL212407B1_D0010.tif" />

where r¹ as defined above for formula XI, with the compound of formula R²X, where R.² as defined above for formula XI and X is a halogen atom or a hydroxy group. In the case where X is a hydroxy group, the reaction was carried out in the presence of activators, preferably an azodicarboxylate e.g. di-tert-butyl azodicarboxylate with a triarylphosphine such as diphenylpyridylphosphine. The reaction may be carried out in an organic solvent, preferably in ether, e.g. dioxane, at a temperature below ambient temperature or preferably at ambient or elevated temperature. The reaction may be carried out using the method described by Mitsonobu, Synthesis 1981, 1 or analogously as further described in the Examples. Compounds of formula XIII are known or can be obtained by known methods.

Compounds of formula VI may be prepared from e.g. (i) benzaldehyde or substituted benzaldehyde according to the method described by Dyke et al., Tetrahedron 1968, 24, 1467 or (ii) from optionally substituted, 1,2-nitrogen protected dihydroisoquinolines by reaction with 2-oxo-carboxylic acid according to the method described by Dyke et al. Tetrahedron 1968, 24, 1467 and the possible subsequent conversion of the obtained carboxylic acid into a methyl ester and then into an alkali metal salt according to the method described by J. March, Advanced Organic Chemistry, 4th Edition, Wiley, New York 1992, pp. 393 and 378 or (iii) from an optionally substituted quinoline or isoquinoline by reaction with a reducing agent (hydride) followed by a 2-oxcarboxylic ester according to the method described by Minter et al., J. Org. Chem., 1988, 53, 2653 or (iv) by introducing substituents into the ring containing the nitrogen atom in an acid of formula VI according to the method described by Janin and Biagani, Terahedron 1993, 39, 10305, or by Ford et al. J. Med . Chem. 1985, 28, 164.

Certain selected compounds of Formula VI can be prepared as follows

(I) reaction scheme

<img file="PL212407B1_D0011.tif" />

^χνι XVII

9 Wherein R, R, R, R, R and R are as defined above. Steps (a) to (c) may be performed according to a known method e.g.

(ii) reaction scheme

<img file="PL212407B1_D0012.tif" />

XXI

9 10 11 12 13 where R.^{4 * * *}. R⁹, R¹⁰, R¹¹, R¹² and r¹³ as defined above. Steps (d) to (g) may be performed according to a known method e.g. step (d) according to the method described by Katayama et al.

Chem. Pharm. Bull., 1980, 28, 2226, step (e) according to the method described by Dyke et al. Tetrahe10

Drone 1968, 24, 1467, steps (f) and (g) according to the method described by J. March, Advanced Organic Chemistry, 4th Edition, Wiley. New York. 1992, pages 393 and 378, or analogously according to the method described hereinafter in the examples;

(iii) reaction scheme

<img file="PL212407B1_D0013.tif" />

for example, a compound of formula XVIII can be reduced with a hydride and then reacted with a 2-oxo-carboxylic acid ester according to the method described by B Minter et al., J. Org. Chem. 1988, 53, 2653, or analogously according to the method described further in the examples;

(iv) for the preparation of compounds of formula VI wherein R.⁵ is a 4-isoquinolinyl group substituted at the 1-position, the following reactions can be performed

<img file="PL212407B1_D0014.tif" />

carried out according to a known method, e.g. steps (h) to (j) according to the method described by Janin and Biagni. Tetrahedron, 1993, 39, 10305 and step (k) according to the method described by J. March, Advaneed Organic Chemistry, 4th Edition, Wiley, New York, 1992, page 378, or analogously according to the method described further in the examples;

(V) to obtain compounds of formula VI wherein R⁵ is an isoquinolinyl group of formula III wherein R⁸ is a cyano group, the following reactions can be performed

<img file="PL212407B1_D0015.tif" />

9 10 11 12 13 where R.⁴, R⁹, R¹⁰, R¹¹, R¹² and r¹³ as defined above. Steps (l) to (n) may be performed according to a known method, e.g., steps (l) and (m) according to the method described by Ford et al., J. Med. Chem., 1985, 28, 164, and step (n) according to the method described by J. March, Advanced Organic Chemistry, 4th Edition, Wiley, New York, 1992, page 378;

₅ (vi) to obtain compounds of formula VI wherein R⁵ is an oxodihydroisoquinolinyl group, the following reactions can be performed

<img file="PL212407B1_D0016.tif" />

9 10 11 12 13 where R.⁴, R⁹, R¹⁰, R¹¹, R¹² and r¹³ as defined above. Steps (o) and (p) can be carried out according to a known method, e.g. according to the method described by Holzgrabe. Arch. Pharm., (Weinheim, Ger.), 1988, 321, 767, or analogously according to the method described further in the examples;

(vii) to obtain compounds of formula VI wherein R.⁵ is a quinolinyl group, the following reactions can be performed

<img file="PL212407B1_D0017.tif" />

according to a known method, e.g. by adding a strong base, preferably an alkali metal dialkylamide e.g.

Of lithium diisopropylamide followed by reaction with carbon dioxide, e.g. according to the method described by Brown and Curless. Tetrahedron Lett., 1986, 27, 6005, or analogously according to the method described further in the examples.

(viii) to obtain compounds of formula VI wherein R.⁵ is a 4-isoquinolinyl group, the following reactions can be performed

<img file="PL212407B1_D0018.tif" />

10 11 12 13 where R.^{8 *}, R¹⁰, R¹¹, R¹² and r¹³ as defined above. Steps (q) to (w) can be performed according to a known method, e.g. step (q) by reacting with a carboxyethyltriarylphosphonium ylide, preferably a carboxyethyltriphenylphosphonium ylide, in an organic solvent, preferably an ether or a hydrocarbon, especially toluene, at a temperature below ambient or elevated temperature, preferably at ambient temperature; step (r) by reaction with nitromethane in the presence of an inorganic base or preferably an amine base, especially tetramethylguanidine in the presence of a solvent

Or preferably in the absence of a solvent at a temperature below ambient temperature, at ambient temperature or preferably at an elevated temperature, e.g. 60-80 ° C; step (s) by reaction with a reducing agent, preferably tin (II) salts, especially tin (II) chloride hydrate in an aqueous or preferably an organic solvent, preferably an alcohol such as ethanol at a temperature below ambient temperature, at ambient temperature or preferably at elevated temperature, e.g. reflux; step (t) by reaction with an acid halide or anhydride, preferably an acid chloride of the acid R⁸COOH at elevated temperature or preferably at a temperature below ambient temperature or at ambient temperature e.g. at a temperature of 0⁰C to ambient temperature, in water or preferably in an organic solvent, especially in a chlorine-containing solvent such as dichloromethane, preferably in the presence of a base e.g. an amine such as triethylamine; step (u) by reaction with a phosphorus halide (V) or an oxyhalide, preferably phosphorus pentachloride or phosphorus oxychloride, preferably in an organic solvent such as a hydrocarbon or nitrile, especially acetonitrile, preferably at ambient temperature or especially at elevated temperature e.g. at reflux temperature; step (v) by reaction with a noble metal, preferably a palladium catalyst, preferably in an organic solvent e.g. in a hydrocarbon such as decalin, preferably at elevated temperature e.g. at boiling point; step (w) by reacting with an alkali metal hydroxide, preferably lithium or sodium hydroxide in an organic solvent, in water or in a water-organic solvent mixture, preferably THF-water at a temperature below ambient temperature, at elevated temperature or preferably at ambient temperature; detailed methods for steps (q) to (w) are further described in the examples.

(ix) reaction scheme

<img file="PL212407B1_D0019.tif" />

where r⁹, R¹¹, R¹² and r¹³ as defined above, Ac is acyl and Y is halogen. Steps (x) to (a) may be carried out using known methods, e.g. in step (x) by reaction with a halogenating agent, e.g.

PL 212 407 B1 succinimide according to the method described in J. March, op. cit., page 531; step (s) by reaction with a reducing agent e.g. a metal hydride in the presence of an acylating agent e.g. acetic anhydride e.g. as described by Katayama et al., op. cit .; step (z) by reaction with a 2-oxo-carboxylic acid, preferably with glyoxylic acid in the presence of a mineral acid e.g. according to the method described by Dyke et al., Tetrahedron 1968, 24, 1467; a reducing agent was used in step (a) as described by J. March et al., op. cit., p. 566, or analogously as further described in the examples.

Certain compounds of formula V are novel compounds, including intermediates 1 to 10 as described below. Certain compounds of formula VI are novel compounds, including intermediates 20 to 48 as described below.

Process variant (b) may be carried out according to a known method, e.g. by reacting a compound of formula I wherein R³ is a hydrogen atom with a suitable alkylating agent, preferably an alkyl iodide or a dialkyl sulfate, with a compound of formula R³J or (R³) 2SO4, where R.³ is a C1-C4-alkyl group. The reaction may be carried out in the presence of an inorganic or organic base, e.g. in water, in an organic solvent or in a water-organic solvent mixture. The alkylation can be carried out below ambient temperature, and most preferably at ambient or elevated temperature. The preferred bases are alkali metal carbonates. The preferred solvents are dipolar aprotic organic solvents, especially N, N-dimethylformamide.

Process variant (c) may be carried out using known sulfonylation methods, e.g. in the presence of an organic or inorganic base, preferably in the presence of a tertiary organic base such as pyridine, at a temperature below ambient temperature, at ambient temperature or preferably at an elevated temperature. The most preferred methods are described below in the examples.

Process variant (d) may be carried out using known methods, e.g. by treating a compound of formula I wherein R² is an alkenyl group, by reaction with a hydroboration agent followed by processing under oxidative-basic conditions. The hydroboration may be conducted below ambient temperature, more preferably at ambient or elevated temperature. Preferred hydroboration agents are dialkylboranes, e.g., 9-borabicyclo [2.2.0] nonane. The reaction is most preferably carried out at reflux temperature. For working up the reaction under oxidative conditions, hydrogen peroxide and an alkali metal hydroxide, preferably sodium hydroxide, are preferably used. The reaction is preferably worked up at a temperature of 40-60 ° C.

Process variant (e) may be carried out using known esterification methods, e.g. by esterifying a compound of formula I, wherein R² is a hydroxy group with a carboxylic acid or a halide, preferably an acid chloride in the presence of an organic or inorganic base at a temperature below ambient temperature, or preferably at ambient or elevated temperature (e.g. at 40-60 ° C). The bases used are tertiary organic bases e.g. pyridine.

Process variant (f) may be carried out using known methods for converting acylamines to amines, e.g. by reaction with a mineral acid, e.g. sulfuric acid or preferably hydrochloric acid. The reaction is preferably carried out in a water-organic solvent mixture, e.g. ethanol-water mixture. The reaction may be carried out at ambient temperature, preferably at elevated temperature, e.g. reflux temperature.

Process variant (g) may be carried out using known dealkylation methods, e.g. by reaction with HBr or with HJ, usually at elevated temperature, preferably by heating with concentrated hydrobromic acid, e.g. as described hereinafter in the Examples.

Process variant (h) may be carried out using known halogenation methods, e.g. by reaction with bromine or chlorine in a solvent such as acetic acid. The reaction is most preferably carried out at ambient temperature as described later in the examples.

Process variant (i) may be carried out using the Simmons Smith reaction method, e.g. reaction with diethylzinc and chloroiodomethane. The reaction is usually carried out in an organic solvent, preferably a halocarbon. The reaction is most preferably run at ambient temperature as described below in the examples.

Compounds of formula I in free form may be converted into salts, and vice versa, in a conventional manner. The free compounds or their salts are obtained in the form of hydrates or solvates containing the solvent used for crystallization. Compounds of formula I in free or salt form may be recovered from the reaction mixture in a conventional manner. A mixture of isomers

The PL 212 407 B1 may be separated into the individual isomers e.g. enantiomers in a conventional manner e.g. by fractional crystallization.

The compounds of formula I in free form or in pharmacologically acceptable salt form, hereinafter referred to as compounds according to the invention, are used for the preparation of a medicament. In particular, they are inhibitors of cyclic guanosine 3 ', 5'-monophosphate phosphodiesterases (cGMP, PDEs), especially PDE5. The compounds of the invention are selective PDE5 inhibitors; they show particularly good selectivity as PDE5 inhibitors compared to inhibitors of other phosphodiesterases, especially PDE1 and PDE6. also showing a low side effect profile.

The medicaments containing the compounds of the present invention have an appropriate duration of action and many of them quickly begin to act as medicaments. Their properties as inhibitors can be determined using the following test:

PDE5 assay: A 10 mM solution of test compound in DMSO was diluted 100 times with 20 vol. aq. DMSO to give 100 µL of a stock solution which was then diluted with 20 vol. v / v of an aqueous solution of DMSO to obtain 10 solutions with a concentration of 10 μΜ to 0.00051 μΜ. 10 µL of each of these solutions was transferred to the selected well of an Optiplate Packard 96-well microtiter dish. To determine total binding, 10 μΙ 20 vol.% the aqueous DMSO solution was added to the other selected wells. To determine non-specific binding, a 10 mM solution of sildenafil in 100% DMSO was diluted 20 times with 20% v / v. aqueous DMSO solution and 10 µl of the solution thus obtained were added to a further selected well of an Optiplate. To all wells containing the test compound solution, aqueous DMSO solution, or sildenafil solution, 80 μl of the test mixture prepared by mixing 2 ml PDE assay buffer, 2 ml aqueous bovine protein (bovine serum albumin) (BSA) containing 5 mg BSA / ml, 2 ml of 75 μl of cGMP sodium salt (0.2 ml), 10 μl of 3H-cGMP (Amersham) and 11.8 ml of distilled water. (PDE assay buffer was prepared by dissolving 7.56 g of Tris base in 800 mL of water, adding 10.325 mL of 1M MgCl2 and 4.25 mL of 0.5M EDTA, adjusting the pH of the solution to 7.5 with 1N hydrochloric acid and diluting to 1 with water. liter). 11 μl of human PDE5 solution, partially purified from platelets. containing 0.017 PDE5 units in 1 μl (1 unit hydrolyzes 1.0 μΜ of 5'-cyclic GMP to 5'-GMP per min at pH 7.5 at 37 ° C), in 20 mM Mepes at pH 7.4 , 100 mM sodium chloride, 10 vol.% glycerol, 1 mM benzamidine and 2 mM dithiotheriethol were diluted 200-fold with enzyme buffer prepared by adding 2 ml of 0.5M EDTA to Tris base solution (1.21 g in 800 ml of water), adjusting the pH of the solution to 7.5 with 1N HCl and diluting with water up to 1 liter. 10 µl of diluted PDE5 solution was added to all wells containing test compound, a mixture of DMSO and water, or sidenafil solution. The plate was incubated for 1 hour. in room temperature. 50 μl of a suspension of 500 mg of Yttrium Silicate SPA (Amersham) beads in 28 ml of water was added to each well and the plate was incubated for a further 20 min and then sealed using a Packard Top Seal-S according to the manufacturer's instructions. The resulting radioactivity results were measured using a Canberra Packard counter (1 minute per well) as a measure of the extent to which binding of PDE5 to the beads is inhibited. The concentration of test compound where 50% inhibition of PDE5 binding to the beads was observed (IC50) was determined in a known manner from the concentration dependence of inhibition.

The compounds in the examples given below have IC50 values ​​ranging from 0.0005 µΜ to 10 µΜ. For example, the compounds in Examples 7, 10, 15, 35, 45, 49, 55, 60, 68, and 70 have IC50 values ​​of 0.007 μΜ, 0.01 μΜ, 0.006 μΜ, 0.010 μΜ, 0.002 μΜ, 0, respectively in the above test. 0037 μΜ, 0.0055 μΜ, 0.0028 μΜ, 0.007 μΜ, and 0.009 μΜ.

In view of inhibiting PDE5, the agents of the invention can be used to treat such metabolic disorders in which PDE5 plays a role. Treatment may be symptomatic or preventive.

The compounds of the invention find use in the manufacture of a medicament for the treatment of disease states associated with pulmonary hypertension and sexual dysfunction, in particular male sexual dysfunction.

The compounds of the invention may be effective in the manufacture of a medicament for the treatment of female sexual dysfunction, the treatment of premature labor, dysmenorrhea, benign prostatic hyperplasia, spasm of the bladder sphincter, urinary or stool incontinence, Prinzmetal variant of coronary artery disease, hypertension, circulatory failure, atherosclerosis, impaired patency of blood vessels, e.g. percutaneous coronary angioplasty, peripheral vascular disease, bronchitis, asthma, allergic rhinitis, tinnitus, bowel movement disorders, e.g. irritable bowel syndrome, eclampsia, Kawasami syndrome, tole16

Nitrates, multiple sclerosis, diabetic peripheral neuropathy, stroke, Alzheimer's disease, acute respiratory failure, psoriasis, skin necrosis, cancer, metastasis, alopecia, esophageal diseases, treatment of anal fissure diseases, and hypoxia due to vascular stenosis.

The compounds according to the invention find use in the preparation of a medicament especially for the treatment of sexual dysfunction and more particularly for the treatment of male erectile dysfunction.

Drugs containing compounds of the invention are combinations of a compound of formula I in free or pharmaceutically acceptable salt form with a pharmaceutically acceptable solvent or carrier. Such combinations are prepared using conventional carriers and methods known in the galenic art. Oral preparations are in the form of tablets and capsules. Formulations for inhalation may be in the form of an aerosol, in other atomized forms, or in the form of a dry powder. Preparations for topical application to the skin are in the form of creams, ointments or gels.

The medicaments containing the compounds of the invention can be used in combination with other PDE5 inhibitors or with other therapeutic agents used in the treatment of sexual dysfunction, in particular male erectile dysfunction, e.g. with alpha-adrenergic receptor antagonists such as phentolamine methane sulfonate with dopaminergic D2 agonists such as such as apomorphine or with NO donors such as L-arginine. The agent of the invention may be mixed with an co-therapeutic agent and may be used in admixture as a pharmaceutical preparation. It can also be used alone before, simultaneously with or after the administration of an adjuvant. The following examples illustrate the invention.

The intermediates of the compound of formula V were obtained as follows:

Interim product 1

Methylallylamine (211 g, 2.97 mol) was added to a solution of concentrated hydrochloric acid (250 mL) in water (1.97 mol) and then potassium cyanate (240 g, 2.97 mol) was added portionwise. The reaction was heated for 2 h. at 80 ° C, then cooled and evaporated to give (2-methyl-allyl) -urea (244.5 g) mp 114-115 ° C. Urea (268 g, 2.35 mol) was added to a solution of cyanoacetic acid (220 g, 2.59 mol) in acetic anhydride (536 ml). The reaction was heated at 70 ° C for 1 h. cooled to 0 ° C and diluted with ether. The resulting precipitate was filtered off, washed with ether, suspended in water and heated to 75 ° C. Then 2M sodium hydroxide solution was added portionwise over 30 min while maintaining the pH in the range 8-9.5. The reaction mixture was cooled to room temperature, 12 ml of acetic acid was added, then cooled to 10 ° C and the precipitate formed was filtered off, washed with cold water and dried to obtain 6-amino-1- (2-methyl-allyl) -1H-pyrimidine- 2,4-dione mp 267-269 ° C. Uracil (253 g, 1.40 mol) was added to a solution of sodium hydroxide (123 g, 3.07 mol) in water (2.5 L). The solution heated up spontaneously and then it was cooled to 20 ° C. Then, in portions within 1 hour. Dimethyl sulfate (196 mL, 2.06 mol) was added. The reaction was left overnight then cooled to 5 ° C, the 6-amino-3-methyl-1- (2-methyl-allyl) -1H-pyrimidine-2,4-dione precipitate was filtered off, mp 162-163 ° C. Methyluracil (165 g, 0.85 mol) was suspended in a mixture of water (1.551) and concentrated hydrochloric acid (72 ml). A solution of sodium nitrite (58.4 g, 0.85 mol) in water (117 ml) was added dropwise to the thus prepared suspension over 30 minutes and stirred for 3 hours. at 20 ° C. The precipitate formed was filtered off, washed successively with water, methanol and ether to give 6-amino-3-methyl-1- (2-methyl-allyl) -5-nitroso-1H-pyrimidine-2,4-dione, mp 213 ° C (with decomposition ). Nitrouracil (190 g, 0.85 mol) was suspended in 950 ml of water, heated to 85 ° C and sodium dithionite (85%, 347.2 g, 1.6 mol) was added portionwise. After cooling to room temperature, the precipitate was filtered to give 5,6-diamino-3-methyl-1- (2-methyl-allyl) -1H-pyrimidine-2,4-dione, mp 152-153 ° C.

Interim product 2

Using the general procedure described for Intermediate 1, (3-nitrobenzyl) urea [J. Med. Chem. 1996, 39, 1924] converted to 6-amino-3-methyl-1- (3-nitrobenzyl) -1H-pyrimidine-2,4-dione [MH]^- 275. A suspension of this compound (4.88 g, 17.7 mmol) in ethanol (200 mL) was hydrogenated with a catalyst 10% Pd / C (0.484 g) 1.5 h. under a pressure of 1 atm. The catalyst was filtered off through a pad of celite, the filtrate was concentrated to obtain 6-amino-1- (3-amino-benzyl) -3-methyl-1H-pyrimidine-2,4-dione [MH] acetate.^- 245. To a suspension of 6-amino-1- (3-amino-benzyl) -3-methyl-1H-pyrimidine-2.4-dione acetate (5.01 g, 16.35 mmol) in 50 ml of pyridine cooled to 0 [Deg.] C, acetic anhydride (1.85 ml, 19.57 mmol) was added. The reaction mixture was warmed to room temperature, stirred for 6 h. and the solvent was evaporated. The residue was triturated with water and the precipitate of N- [3- (6-amino-3-methyl-2,4-dioxo-3,4-dihydro-2H-pyrimidin-1-ylmethyl) -phenyl] acetamide was filtered off.¹H NMR (400 MHz. DMSO): δ: 2.00 (s 3H), 3.09 (s 3H), 4.72 (s 1H), 5.02 (s 2H), 6.75 (s 2H) ,

PL 212 407 B1

6.88 (d J 6 1H), 7.25 (t J 6 1H), 7.30 (s 1H), 7.55 (d J 6 1H). Using the general procedure for Intermediate 1, this compound was converted to N- [3- (5,6-diamino-3-methyl-2,4-dioxo-3,4-dihydro-2H-pyrimidin-1-ylmethyl) - phenyl] acetamide [MH]⁺ 304.

Interim product 3

Using the general procedure described for Intermediate 1, (4-nitrobenzyl) urea [J. Med. Chem. 1996, 39, 1924] converted to 6-amino-3-methyl-1- (4-nitro-benzyl) -1H-pyrimidine-2,4-dione [MH]⁺ 277. A solution of calcium chloride (4.94 g, 45 mmol) in water (100 ml) was added to the solution of 6-amino-3-methyl-1- (4-nitro-benzyl) -1H-pyrimidine-2,4-dione (19.08 g, 69.0 mmol) in acetic acid (300 mL). Zinc dust (58.8 g, 900 mmol) was then added portionwise while cooling the reaction mixture, then stirred at room temperature for 1.5 h, filtered through a celite pad and washed successively with ethanol and acetic acid. After evaporation of the filtrate and washings, 6-amino-1- (4-amino-benzyl) -3-methyl-1H-pyrimidine-2,4-dione acetate [M-3H] was obtained.243. To a 0 ° C-cooled suspension of 6-amino-1- (4-aminobenzyl) -3-methyl-1H-pyrimidine-2,4-dione (17.0 g, 69.0 mmol) in 260 mL of pyridine was added ( 7.2 mL, 76.0 mmol) of acetic anhydride. The reaction mixture was warmed to room temperature, stirred for 6 h. and the solvent was evaporated. The residue was triturated with water, filtered off and the resulting precipitate of N- [4- (6-amino-3-methyl-2,4-dioxo-3,4-dihydro-2H-pyrimidin-1-ylmethyl) -phenyl] -acetamide [ MH]⁺ 289. Using the general procedure described for Intermediate 1, this compound was converted to N- [4- (5,6-diamino-3-methyl-2,4-dioxo-3,4-dihydro-2H-pyrimidin-1-ylmethyl). ) -phenyl] acetamide M [MH]⁺ 304.

Interim product 4

To a 0 ° C-cooled suspension of 6-chloromethyluracil (2.0 g, 12 mmol) in THF / dioxane (1: 1, 16 ml) was added 2-pyridyl diphenylphosphine (3.60 g, 13.7 mmol) followed by cyclobutanomethanol (1.3 mL, 13.8 mmol) then added ditertbutyl azodicarboxylate (3.15 g, 13.7 mmol). The mixture was stirred overnight at room temperature, then 15 ml of a 4M solution of hydrogen chloride in dioxane was added and evaporated. The residue was dissolved in methylene chloride, washed with 3.5M hydrochloric acid and dried with magnesium sulfate. After evaporation of methylene chloride, the crude product was purified by flash chromatography, eluent: methylene chloride - methanol (100: 1). Obtained 6-chloro-1-cyclobutylmethyl-3-methyl-1H-pyrimidine-2,4-dione,¹H NMR (400MHz. CDCl3): δ: 1.702.00 (m 6H), 2.60 (m 1H), 3.20 (s 3H), 4.00 (d J 7 2H), 5.78 (s 1H ) which was dissolved in 50 ml of n-butanol and added (4 ml, 26.5 mmol) of veratrylamine and heated for 16 h. at boiling point. The solvent was evaporated, the residue was dissolved in methylene chloride, washed with 1M aqueous hydrochloric acid solution, dried with magnesium sulfate and the methylene chloride was evaporated. The crude product was purified by flash chromatography, eluent: methylene chloride - methanol (50: 1) to give 1-cyclobutylmethyl-6- (3,4-dimethoxy-benzylamino) -3-methyl-1H-pyrimidine-2,4-dione ¹H NMR (400MHz, CDCl3) d 1.60-1.80 (m 4H), 1.80-2.00 (m 2H), 2.50 (m 1H), 3.21 (s 3H), 3. 80 (s 6H), 3.85 (d J 7 2H), 4.11 (d J 5 1H), 4.25 (m 1H), 4.84 (s 1H), 6.74 (s 1H), 6.80 (s, 2H), which was dissolved in 50 mL of formic acid, was added 0.26 g of field black and heated at 40 ° C for 21 hours. It was then filtered through Celite, evaporated and purified using preparative HPLC. Obtained 6-amino-1-cyclobutylmethyl-3-methyl-1H-pyrimidine-2,4-dione M⁺209, which was converted to 5,6-diamino-1-cyclobutylmethyl-3-methyl-1H-pyrimidine-2,4-dione using the general method described for Intermediate 1. HPLC retention time 0.17 min (4 min gradient acetonitrile - water 30-95%).

Interim product 5

5,6-diamino-3-methyl-1- (tetrahydrofuran-2-ylmethyl) -1H-pyrimidine-2,4-dione mp 115-116 ° C prepared from (tetrahydrofuran-2-ylmethyl) -urea (Collect. Czech. Chem. Commun. 1972, 37, 2786) using the general method described for Intermediate 1.

Interim product 6

5,6-diamino-3-methyl-1- (2-methylbutyl) -1H-pyrimidine-2,4-dione mp 163-165 ° C was prepared using the general method described for Intermediate 1.

Interim product 7

5,6-diamino-1-hexyl-3-methyl-1H-pyrimidine-2,4-dione was prepared from 6-amino-1-hexyl-1H-pyrimidine-2,4-dione (J. Med. Chem. 1993 36, 1465) using the general method described for Intermediate 1, HPLC retention time 6.01 (8 min acetonitrile - water gradient 0-95%).

PL 212 407 B1

Interim product 8

5.6-diamino-1- (3,4-dimethoxy-benzyl) -3-methyl-1H-pyrimidine-2,4-dione [MH]^- 305 was prepared from (3,4-dimethoxybenzyl) urea (Farmaco, Ed. Sci. 1977, 32, 813) using the general method described for Intermediate 1.

Interim product 9

5.6-diamino-1-benzo [1,3] dioxol-5-ylmethyl-3-methyl-1H-pyrimidine-2,4-dione mp 183-186 ° C was prepared using the general method described for Intermediate 1.

Interim product 10

5.6-diamino-1- (2,4-dichloro-benzyl) -3-methyl-1H-pyrimidine-2,4-dione was prepared using the general method described for Intermediate 1. ¹H NMR (400MHz, DMSO-d6) δ 3.16 (s 3H), 5.05 (s 2H), 6.18 (s 2H), 6.82 (d, J 9 1H), 7.38 (d , J 9 1H), 7.62 (s 1H).

Other intermediates of the compound of formula V were prepared according to the literature listed below:

Literature:

(1) Eur. J. Med. Chem. 1990, 25, 653 (2) J. Med. Chem. 1996, 39, 2 (3) FR 2 531 085 (4) Eur. J. Med. Chem. -Chim. Ther. 1974,9,313

<img file="PL212407B1_D0020.tif" />

Interim product 20

A mixture (0.50 g, 1.68 mmol) of 3- (3,4-dimethoxy-phenyl) -5-nitropentanoic acid ethyl ester [J. Med. Chem., 1989, 32, 1450] and (1.90 g, 8.4 mmol) hydrated tin (II) chloride in 10 mL of ethanol were heated for 2 h. at reflux, then cooled to room temperature and evaporated. The crude product was dissolved in 15 mL of methylene chloride, cooled to 0 ° C, and 5 mL of triethylamine was added followed by 3,5-dimethoxybenzoyl chloride (0.404 g, 2.02 mmol). The mixture was stirred overnight at room temperature, then evaporated and dissolved in ethyl acetate, washed with water and dried with sodium sulfate. Ethyl acetate was evaporated and the crude product was purified by flash chromatography; eluent: hexane - ethyl acetate (1: 1). 4- (3,5-Dimethoxy-benzoylamino) -3- (3,4-dimethoxy-phenyl) -butyric acid ethyl ester [MH] was obtained.⁺ 432. This compound (0.200 g, 0.46 mmol) was dissolved in 8 mL of acetonitrile, and phosphorus oxychloride (0.211 g, 1.38 mmol) was added.

PL 212 407 B1 and heated for 3 hours. at boiling point. The solvent was then evaporated, the residue was dissolved in ethyl acetate, washed with saturated sodium carbonate solution, dried with sodium sulfate, and ethyl acetate was evaporated to obtain [1- (3,5-dimethoxy-phenyl) -6.7-dimethoxy- acid ethyl ester. 3,4-dihydro-isoquinolin-4-yl] -acetic [MH]⁺ 414. This compound (0.50 g, 1.21 mmol) was dissolved in 10 ml of decoline and 50 mg of 10% Pd / C was added and heated for 2.5 h. at 190 ° C, then cooled to room temperature and methylene chloride was added. The solution was filtered through Celite, the filtrate and washings evaporated to give [1- (3,5-dimethoxyphenyl) -6,7-dimethoxy-isoquinolin-4-yl] -acetic acid ethyl ester.¹H NMR (400MHz, CDCl3) δ 1.18 (t J 7 3H), 3.78 (s 6H), 3.80 (s 3H), 3.90 (s 2H), 3.99 (s 3H), 4, 10 (q J 7 2H), 6.50 (d J 0.5 1H), 6.75 (d J 0.5 2H), 7.20 (s 1H), 7.36 (s 1H), 8, 40 (s, 1H). This compound (0.30 g, 0.73 mmol) was dissolved in 10 mL of methanol, added (0.80 mL, 0.80 mmol) 1M aqueous lithium hydroxide solution and stirred overnight at room temperature. After evaporation of the methanol, the pH of the residue was adjusted to 7 by adding 1M aqueous hydrochloric acid. The resulting precipitate was filtered off, and after drying, [1- (3,5-dimethoxy-phenyl) -6,7-dimethoxy-isoquinolin-4-yl] -acetic acid was obtained.

Interim product 21

A mixture of 3-isopropoxy-4-methoxy-benzaldehyde (3.9 g, 20 mmol) and (ethoxycarbonylmethylene) triphenylphosphate (6.96 g, 20 mmol) in 100 ml of toluene was heated at reflux for 2 hours, cooled to temperature room and evaporated. The crude product was dissolved in methylene chloride and filtered through a pad of silica gel to give (E) -3- (3-isopropoxy-4-methoxyphenyl) -acrylic acid ethyl ester TLC Rf 0.70; hexane - ethyl acetate (1: 1). This acid was dissolved in 10 ml of nitromethane, 0.5 ml of 1,1,3,3-tetramethylguanidine was added and the mixture was heated for 36 hours. at 70 ° C. The solvent was evaporated, the residue was dissolved in ethyl acetate and washed with a 2N aqueous solution of hydrochloric acid, then with water and brine. After drying with sodium sulfate and evaporation of ethyl acetate, the obtained crude product was purified by flash chromatography; eluent: hexane - ethyl acetate (4: 1) to obtain 3- (3-isopropoxy-4-methoxy-phenyl) -4-nitro-butyric acid ethyl ester,¹H NMR (400MHz, CDCl3) δ 1.20 (t, J 7 3H), 1.38 (d J 7 6H), 2.75 (d J 6 2H), 3.90 (m 1H), 4.10 (m 2H), 4.48-4.78 (m 3H), 6.75-6.86 (m 3H). This compound was converted to [1- (3,5-diisopropoxyphenyl) -6-isopropoxy-7-methoxy-isoquinolin-4-yl) -acetic acid using the general method described for Intermediate 20. Identified as ethyl ester [MH]⁺ 496.

Interim product 22

To 2,2-dimethoxyethylamine (21 mmol) was added a solution of 3-ethoxy-4-methoxy-benzaldehyde (3.6 g, 20 mmol) in 15 mL of ethanol and heated to reflux for 2 h. The reaction mixture was then cooled to room temperature, sodium borohydride (0.794 g, 21 mmol) was added and stirred overnight at room temperature. Ethanol was distilled off, water was added and extraction was carried out with ethyl acetate. The combined organic extracts were washed with water, brine and dried with magnesium sulfate, the solvent was evaporated to give (2,2-dimethoxy-ethyl) - (3-ethoxy-4-methoxybenzyl) amine [MH]⁺ 270. This compound (2.70 g, 10 mmol) was suspended in 50 ml of 6N hydrochloric acid and glyoxolic acid (0.88 g, 12 mmol) was added and the mixture was heated for 1 h. at 100 ° C. After cooling to room temperature, 30 ml of methanol was added, the mixture was filtered and identified as the methyl ester M⁺ 276. Lithium hydroxide (10 mmol) in a THF-methanol-water mixture was added to the filtrate and left to stand overnight. After distilling off the solvents, the crude product was partitioned between water and methylene chloride. The aqueous layer was washed with methylene chloride and the solvent was evaporated to dryness to give (7-ethoxy-6-methoxy-isoquinolin-4-yl) -acetic acid lithium salt which was used to prepare xanthine without identification.

Interim product 23

To a solution of (6,7-dimethoxy-isoquinolin-4-yl) -acetic acid ethyl ester (Tetrahedron 1973, 29, 3881) (1.668 g, 6.07 mmol) in 20 ml of chloroform was added portionwise over 5 hours. m-chloroperbenzoic acid (1.153 g, 6.67 mmol). The reaction mixture was washed with saturated sodium hydrogen carbonate solution and brine, dried with MgSO4, and the solvent was distilled off to obtain (6,7-dimethoxy-2-oxo-isoquinolin-4-yl) -acetic acid ethyl ester 1.71 g. 96%. The ester was dissolved in 30 mL of chloroform, POCl3 (3 mL, 32.3 mmol) was added and heated for 2 h. At boiling point. After the chloroform was evaporated, methylene chloride and ice water were added and the mixture was made basic with an aqueous ammonia solution. The aqueous layer was then extracted with methylene chloride, the combined organic layers were washed with brine, dried with magnesium sulfate, and evaporation of the solvent gave ethyl ester of (1-chloro-6,7-dimethoxy-isoquinolin-4-yl) -acetic acid. Chloroester (0.50 g, 1.6 mmol) was suspended in 15 mL of 2M sodium hydroxide, added

Ml of ethanol and the solution was stirred for 2 hours. at room temperature and the solvent was distilled off. Upon acidification to pH 2 with concentrated hydrochloric acid, a precipitate formed which was filtered off and dried to obtain (1-chloro-6,7-dimethoxy-isoquinolin-4-yl) -acetic acid.¹H NMR (400MHz, DMSO-d6) δ: 3.92 (s 3H), 3.96 (s 3H), 4.02 (s 2H), 7.31 (s 1H), 7.44 (s 1H) , 8.04 (s, 1H).

Interim product 24

Pyruvic acid was added to (2,2-dimethoxy-ethyl) - (3-methoxy-benzyl) amine (Tetrahedron, 1973, 29, 3881) and reacted according to the general procedure for Intermediate 22 to give the hydrochloride of the acid 2- (7- methoxy-isoquinolin-4-yl) propionic acid, mp 174-176 ° C. After reaction with gaseous hydrogen chloride in ethanol, the hydrochloride of the corresponding ethyl ester was obtained, m.p. 190-192 ° C, which was then reacted sequentially with m-chloroperbenzoic acid and phosphorus oxychloride according to the procedure described for Intermediate 23 to obtain 2- (1-chloro-7-methoxy-isoquinolin-4-yl) -propionic acid ethyl ester mp 126-128 ° C. This compound (47 g, 0.16 mol) was dissolved in 400 ml of ethanol, 150 ml of 2N sodium hydroxide were added, the mixture was heated for 1 hour. at 60 ° C and the solvent was distilled off. After crystallization from acetone, 2- (1-chloro-7-methoxy-isoquinolin-4-yl) -propionic acid was obtained, mp 167-168 ° C.

Interim product 25

Dimethyl sulfate (12.7 ml, 0.10 mol) was added portionwise to a solution of 2- (7-methoxy-2-oxy-isoquinolin-4-yl) -propionic acid ethyl ester (28 g, 0.10 mol). The reaction temperature was increased to 100 ° C and this temperature was maintained for 2 hours. It was then cooled to room temperature, dissolved in 50 ml of water, and a solution of sodium cyanide (15 g, 0.31 mol) in 90 ml of water was added over 10 minutes while cooling the reaction mixture and stirred for 3 hours. in room temperature. The crude product was extracted with chloroform, the chloroform extracts were washed with saturated sodium bicarbonate solution, brine, dried with sodium sulfate and the solvent was evaporated. After crystallization from 3N ethanolic HCl in ether, 2- (1-cyano-7-methoxy-isoquinolin-4-yl) -propionic acid ethyl ester hydrochloride with mp 89-98 ° C was obtained. This compound was made acidic by hydrolysis as described for Intermediate 22 and used for xanthine synthesis without purification.

Interim product 26

To a solution of isoquinoline (1.64 g, 12.7 mmol) in 25 mL of THF was added dropwise a solution of sodium triethylborohydride (1M THF, 12.7 mL, 12.7 mmol). The reaction was stirred at room temperature for 1 hr. then a solution of ethyl glyoxalate (1.43 g, 13.9 mmol) in toluene (previously heated 1.5 h at 110 ° C) was added dropwise. Then it was stirred for 4 hours. at room temperature, cooled to 0 ° C and 25.4 ml of a 0.5M aqueous solution of sodium hydroxide, 12.7 ml of a 30% aqueous solution of hydrogen peroxide were added and stirred for 2 hours. Acidified with 1N HCl, washed three times with ethyl acetate; the aqueous layer was concentrated and cooled in the refrigerator. The resulting precipitate was filtered off and dried to give isoquinolin-4-yl-acetic acid hydrochloride MH⁺ 188.

Interim product 27

To (1-chloro-6,7-dimethoxy-isoquinolin-4-yl) -acetic acid ethyl ester (0.200 g, 0.65 mmol) suspended in 1 ml of toluene was added excess morpholine and heated to reflux until the starting material disappeared completely. . After evaporation of the solvent, the residue was partitioned between water and methylene chloride; the organic layer was separated, dried with magnesium sulfate, and the solvent was evaporated to obtain (6,7-dimethoxy-1-morpholin-4-yl-isoquinolin-4-yl) -acetic acid ethyl ester [MH]⁺ 361. The crude ester (0.240 g, 0.66 mmol) was dissolved in 20 mL of ethanol, 3 mL of 2M sodium hydroxide solution was added and stirred overnight at room temperature. Concentrated hydrochloric acid was then added to adjust the pH to 1, the solvent was evaporated and the crude acid was used without purification for the synthesis of Xanthine Intermediate.

Interim product 28

The procedure of Intermediate 27 was repeated using an excess of N-methylpiperazine in place of morpholine to give [6,7-dimethoxy-1- (4-methyl-piperazin-1-yl) -isoquinolin-4-yl] -acetic acid ethyl ester (0.186 g , 38%) ¹H NMR (DMSO-d6) δ 1.19 (t J 7 3H), 2.30 (s 3H), 2.61 (m 4H), 3.10-3.30 (m 4H), 3.92 ( s 6H), 3.97 (s 2H), 4.10 (q J 7 2H), 7.19 (s 1H), 7.37 (s 1H), 7.91 (s 1H). The ester (0.186 g, 0.50 mmol) was dissolved in 20 mL of ethanol, 3 mL of 2M sodium hydroxide solution was added and stirred overnight at room temperature. After adjusting the pH of the solution to 1 with concentrated hydrochloric acid, the solvent was distilled off and the crude acid was used without purification for the synthesis of Xanthine Intermediate.

PL 212 407 B1

Interim product 29

To a solution of 6-methoxyisoquinoline (Synth. Commun. 1999, 29, 1617) (0.207 g, 1.30 mmol) in 9 mL of acetic acid, N-chlorosuccinimide (0.347 g, 2.60 mmol) was added. The reaction was heated for 3 h. at 50 ° C, cooled to room temperature, evaporated and the residue was partitioned between ethyl acetate and 1M aqueous sodium hydroxide solution. The organic layer was washed with water and brine, dried with magnesium sulfate and the ethyl acetate evaporated to give 5-chloro-6-methoxyisoquinoline [MH]⁺ 194. To a solution of this intermediate (0.175 g, 0.90 mmol) in 4.5 mL of THF was added acetic anhydride (0.101 mL, 1.08 mmol) and sodium triacetoxyborohydride (0.229 g, 1.08 mmol) and stirred for 22 at at ambient temperature. The solvent was then evaporated, the residue was dissolved in ethyl acetate, washed with 0.5M aqueous hydrochloric acid, brine and dried with magnesium sulfate. After evaporation of the solvent, 1- (5-chloro-6-methoxy-1H-isoquinolin-2-yl) -ethanone was obtained, m.p. 78-80 ° C. A suspension of this compound (0.150 g, 0.60 mmol) and glyoxylic acid (76 mg, 0.80 mmol) in 6M aqueous hydrochloric acid (2.8 mL) was heated at 100 ° C for 3 h. After cooling to room temperature, the resulting precipitate was filtered off to obtain (5-chloro-6-methoxy-isoquinolin-4-yl) -acetic acid [MH]⁺ 252. To a suspension of this compound (0.970 g, 3.38 mmol), ammonium formate (1.05 g, 16.9 mmol) in 25 mL of acetic acid - water 1: 1 was added 10% Pd / C (0.730 g ) and stirred for 16 hours. at ambient temperature. It was then filtered through Celite, the filtrate and washings were evaporated and purified by extraction with acetone in a Soxhlet apparatus. (6-Methoxy-isoquinolin-4-yl) -acetic acid was obtained. Another method of reducing (5-chloro-6-methoxy-isoquinolin-4-yl) -acetic acid to (6-methoxy-isoquinolin-4-yl) -acetic acid is by mixing a suspension of (5-chloro-6-methoxy- isoquinolin-4-yl) acetic acid (20 g, 69.4 mmol) in 400 ml of 1M sodium hydroxide solution for 20 min, filtering the salt formed and reducing with hydrogen against 10% Pd / C (1.4 g) at atmospheric pressure for 2.25 hours The resulting suspension was filtered through glass wool and celite and washed with water. After filtration, the solution was cooled in an ice bath and slowly neutralized over 30 min, then acidified with 5M hydrochloric acid (80 ml). The resulting suspension turns into crystals after 20 hours. at 5 ° C, The crystals were filtered off, washed with cold ethanol (25 ml) and dried under reduced pressure. (6-Methoxy-isoquinolin-4-yl) -acetic acid was obtained.

Interim product 30

To a mixture of 1- (6-chloro-1H-isoquinolin-2-yl) -ethanone (J. Org. Chem., 1980, 45, 1950) (1.44 g.

5.90 mmol) in 6N HCl (24 mL) was added glyoxylic acid (1.37 g, 9.28 mmol). The reaction was heated for 3 h. at 100 ° C, cooled to room temperature, washed with ether and evaporated to a volume of 10 ml. It is cooled overnight in a refrigerator and the precipitate formed is filtered off and dried to obtain (6-chloro-isoquinolin-4-yl) -acetic acid hydrochloride.¹H NMR (400 MHz, DMSO) δ: 4.45 (s 2H), 8.18 (d J 9 1H), 8.52 (s 1H), 8.70 (d, J 8 1H), 8.83 (s 1H), 9.96 (s 1H).

Interim product 31

To a 0 ° C-cooled solution of 6-bromoisoquinoline (J. Chem. Soc. Perkin Trans 2, 1998, 437) (1.544 g, 7.42 mmol) in 10 ml acetic acid and 3 ml acetic anhydride was added sodium borohydride ( 1.12 g, 29.6 mmol). After 4 hours heating at 60 ° C, the reaction mixture was cooled, evaporated and diluted with water. The mixture was adjusted to pH 10 with potassium carbonate, extracted with ethyl acetate, the organic layer was washed twice with 0.5N HCl and brine, and dried with sodium sulfate. Evaporation gave 1- (6-bromo-1H-isoquinolin-2-yl) -ethanone MH⁺253. To a mixture of 1- (6-bromo-1H-isoquinolin-2-yl) -ethanone (1.50 g, 5.90 mmol) in 20 mL of 6N HCl was added glyoxylic acid (0.812 g, 8.80 mmol) and heated for 2 hours. at 100 ° C, then cooled to room temperature and washed with ethyl acetate. After evaporation, the residue was dissolved in 20 ml of methanol, 10 drops of concentrated sulfuric acid were added and heated for 14 hours. at boiling point. After partial evaporation of the solvent, the precipitate formed was filtered off, washed with methanol and dried to obtain (6-bromo-isoquinolin-4-yl) -acetic acid methyl ester hydrochloride, MH⁺ 281. To a 0 ° C solution of the obtained (6-bromo-isoquinolin-4-yl) -acetic acid methyl ester hydrochloride (50 mg, 0.18 mmol) dissolved in 3 mL THF - water (3: 1) lithium hydroxide hydrate was added. After 1 hour the solvent was evaporated to obtain the lithium salt of (6-bromo-isoquinolin-4-yl) -acetic acid MH⁺ 266.

Interim product 32

To a suspension of (6-bromo-isoquinolin-4-yl) -acetic acid methyl ester (0.325 g, 1.03 mmol) in 1.75 mL DMF was added trimethylsilylacetylene (0.17 mL, 1.23 mmol), triethylamine (10 mL), cuprous iodide (40 mg, 0.20 mmol) and (Ph3P) 2PdCl2 (73 mg, 0.10 mmol). Everything was heated for

The reaction time was 212 407 B1 min at 45 ° C, cooled to ambient temperature and diluted with ethyl acetate. After washing with water and brine, the organic layer was dried with magnesium sulfate, the solvent was evaporated and the crude product was purified by flash chromatography (eluent: hexane - ethyl acetate 1: 1). Obtained (6-trimethylsilanylethynyl-isoquinolin-4-yl) -acetic acid methyl ester [MH]⁺298. This compound (0.221 g, 0.74 mmol) was dissolved in 7.5 mL of methanol and potassium carbonate (75 mg, 0.54 mmol) was added. The reaction was stirred for 30 min at ambient temperature, the solvent was evaporated, the residue was purified by flash chromatography (eluent: methylene chloride - methanol 5: 1) to obtain (6-ethynyl-isoquinolin-4-yl) -acetic acid [MH]⁺ 212.

Interim product 33

To a 0 ° C-cooled solution of 6-methoxyisoquinoline (Synth. Commun. 1999, 29, 1617) was added bromine (0.211 ml, 6.28 mmol) in 10 ml methylene chloride and stirred for 20 h. at ambient temperature. The reaction mixture was then poured into 1M aqueous sodium hydroxide solution, the layers were separated, the organic layer was washed with brine, dried with magnesium sulfate and the solvent was evaporated. The crude product was purified by flash chromatography (eluent: methylene chloride - methanol 20: 1) to give 5-bromo-6-methoxyisoquinoline [MH]⁺ 240, converted to (5-bromo-6-methoxy-isoquinolin-4-yl) -acetic acid [MH]⁺ 298 according to the method described for Intermediate 29.

Interim product 34

[1- (3,5-Diisopropoxy-phenyl) -6,7-dimethoxy-isoquinolin-4-yl) -acetic acid was prepared according to the method described for Intermediate 20. ¹H NMR (400MHz CDCl3) δ 1.25 (d, J612H), 3.78 (s 3H), 3.86 (s 2H), 3.92 (s 3H), 6.46 (d J 0, Δ 1H), 6.65 (d J 0.5 2H), 7.20 (s 2H), 8.30 (s 1H).

The following compounds were prepared according to the method described for Intermediate 21:

Interim product 35

1- (3,5-Dimethoxy-phenyl) -6-isopropoxy-7-methoxy-isoquinolin-4-yl) -acetic acid, [MH]⁺ 412.

Interim product 36

(1-tert.-butyl-6-isopropoxy-7-methoxy-isoquinolin-4-yl) -acetic acid, ¹H NMR (400MHz, CDCl3) δ 1.32 (d J 7 6H), 1.52 (s 9H), 3.80 (s 2H), 3.90 (s 3H), 4.75 (heptet J 7 1H ), 7.28 (s 1H), 7.66 (s 1H), 8.08 (s 1H).

Interim product 37

(6-isopropoxy-1-isopropyl-7-methoxy-isoquinolin-4-yl) -acetic acid, [MH] ⁺ 318.

The following compounds were prepared according to the method described for Intermediate 20:

Interim product 38

(6,7-Dimethoxy-1-methyl-isoquinolin-4-yl) -acetic acid, [MH]⁺ 262.

Interim product 39

(1-tert.-Butyl-6,7-dimethoxy-isoquinolin-4-yl) -acetic acid, ¹H NMR (400MHz, CDCl3) d 1.75 (s 9H), 3.95 (s 6H), 4.04 (s 2H), 7.28 (s 1H), 7.75 (s 1H), 8, 66 (s, 1H).

Interim product 40

(1-isopropyl-6,7-dimethoxy-isoquinolin-4-yl) -acetic acid, ¹H NMR (400MHz, CDCl3) δ 1.25 (t J 7 3H), 1.45 (d J 7 3H), 3.82 (J 7 1H heptet), 3.90 (s 2H), 3.08 ( s 2H), 4.15 (q J 7 2H), 7.28 (s 1H), 7.48 (s 1H), 8.30 (s 1H).

Interim product 41

2- (7-Methoxy-1-morpholin-4-yl) -propionic acid mp 225-227 ° C, obtained according to the procedure for Intermediate 27.

The following compounds were prepared analogously to Intermediate 22:

Interim product 42

[MH]⁺ 332 (7-Hydroxy-6-methoxy-isoquinolin-4-yl) -acetic acid, lithium salt, via (3-benzyloxy-4-methoxy-benzyl) - (2,2-dimethoxy-ethyl) -amine.

Interim product 43

(6,7-Dimethoxy-3-methyl-isoquinolin-4-yl) -acetic acid, ¹H NMR (400MHz, DMSO) δ: 2.50 (s 3H), 3.91 (s 3H), 3.93 (s 3H), 4.02 (s 2H), 7.30 (s 1H), 7 . 43 (s 1H), 8.30 (s 1H).

Interim product 44

(6-ethoxy-7-methoxy-isoquinolin-4-yl) -acetic acid, M⁺ 261.

The following compounds were prepared analogously to Intermediate 22 using pyruvic acid in place of glyoxylic acid:

PL 212 407 B1

Interim product 45

2- (6-ethoxy-7-methoxy-isoquinolin-4-yl) -propionic acid lithium salt, characterized as methyl ester, M⁺ 290.

Interim product 46

2- (7-ethoxy-6-methoxy-isoquinolin-4-yl) -propionic acid lithium salt, characterized as methyl ester, M⁺ 290.

Interim product 47

6,7- (dimethoxy-isoquinolin-4-yl) -propionic acid, characterized as a methyl ester. M.⁺ 276.

Interim product 48

(8-Fluoro-6-methoxy-isoquinolin-4-yl) -acetic acid was prepared according to the procedure of Intermediate 31 and characterized as the methyl ester, [M]⁺ 250.

Interim product 49

(6,7-Dimethoxy-isoquinolin-4-yl) -acetic acid and Intermediate 50: [1,3] dioxolo [4,5-g] isoquinolin-8-yl-acetic acid, was prepared as described in Dyke et al. .. Tetrahedron 1968, 24, 1467.

Interim product 51

(7-Methoxy-isoquinolin-4-yl) -acetic acid was prepared according to Dyke et al., Tetrahedron, 1973, 29, 3881.

Interim 52

2,2-Dimethoxyethylamine (13.85 ml, 0.13 mol) was added to a solution of 3-fluoro-4-methoxybenzaldehyde (20 g, 0.13 mol) in toluene (200 ml). The resulting solution was purged with nitrogen and refluxed overnight in a Dean-Stark apparatus. After removing the solvent under reduced pressure, (2,2-dimethoxy-ethyl) - [1- (3-fluoro-4-methoxyphenyl) methylidene] amine was obtained, which was dissolved in ethyl acetate and acetic anhydride (13.1 g, 0.13 mol). Platinum oxide was added under an atmosphere of nitrogen and the mixture was stirred under an atmosphere of hydrogen until absorption was complete. After filtration, washing with saturated aqueous NaHCO3 (3 x 100 mL), brine and water, drying over MgSO4 and concentration, N- (2,2-dimethoxy-ethyl) -N- (3-fluoro-4-methoxy-benzyl) - acetamide. This compound (38.9 g, ca 0.13 mol) was dissolved in anhydrous CH2Cl2 and added slowly over 20 min with stirring to a mixture of AlCl3 (90 g) and CH2Cl2 under a nitrogen atmosphere. The total volume of CH2Cl2 was 250 ml. The mixture was stirred for a further 10 min at room temperature and 40% aqueous NaOH was added while cooling in an ice bath. The mixture was then diluted with water (250 mL), filtered through glass wool, the organic phase was separated and the aqueous phase was extracted with CH2Cl2 (2 x 200 mL). After drying over MgSO4 and evaporation under reduced pressure, a crude oil was obtained which was purified by flash chromatography (eluent 1% methanol in CH2Cl2) to give 1- (7-fluoro-6-methoxy-1H-isoquinolin-2-yl) -ethanone as one with products. This compound (0.60 g, 2.7 mmol) was mixed with glyoxylic acid (0.325 g, 3.5 mmol) and water (10 ml) and stirred for 20 min at room temperature. Concentrated hydrochloric acid (10 ml) was added and the resulting mixture was heated under reflux for 1 h. Concentration and purification by preparative HPLC gave (7-fluoro-6-methoxy-isoquinolin-4-yl) -acetic acid, [MH]⁺ 236.

The following compounds were prepared analogously to Intermediate 20:

Interim product 53

(1-Methyl-6-methoxy-isoquinolin-4-yl) -acetic acid.

Interim product 54

(6-Isopropoxy-1-methyl-isoquinolin-4-yl) -acetic acid.

Interim product 55

(6-Ethoxy-1-methyl-isoquinolin-4-yl) -acetic acid.

Interim product 56

A solution of (6-bromo-isoquinolin-4-yl) -acetic acid methyl ester (52 mg, 0.19 mmol), obtained as an intermediate for Intermediate 31, in DMF (3 ml) was added to zinc dicyanide (26 mg, 0.22 mmol) under a nitrogen atmosphere. 1,1'-Bis (diphenylphosphino) ferrocene (15 mg) and tris (dibenzylideneacetone) dipalladium (0) (8 mg) were added to the resulting mixture, and the mixture was stirred at 120 ° C for 22 hours. The solution was cooled and diluted with chloroform (30 ml), washed with water (2 x 20 ml) and brine (20 ml). Chloroform (40 mL) was added again, and the solution was dried over MgSO4, filtered and concentrated. Two flash column chromatography on silica gel (eluents 40: 1 CH2Cl2: methanol then 50: 1 CH2Cl2: methanol) gave (6-cyano-isoquinolin-4-yl) -acetic acid methyl ester [MH]⁺ 227. This compound was saponified with LiOH in 3: 1 THF / water. The resulting mixture was partially evaporated to remove THF, diluted with water

The mixture was added to 10 ml and washed with ethyl acetate. The aqueous phase was neutralized with 1M hydrochloric acid (to pH 4-5) and extracted exhaustively with ethyl acetate. The organic phase, after drying over MgSO4, was filtered and concentrated to give (6-cyano-isoquinolin-4-yl) -acetic acid M⁺ 212.

Interim product 57

(5-Chloro-6-methoxy-isoquinolin-4-yl) -acetic acid was obtained as described in the procedure for Intermediate 29.

Interim product 58

K2CO3 (72 mg) was added to a solution of (6-trimethylsilanylethynyl-isoquinolin-4-yl) -acetic acid methyl ester prepared as in Intermediate 32 (0.19 g, 0.64 mmol) in anhydrous methanol (7 ml). and it was stirred for 1 hour. Another portion of K2CO3 (11 mg) was added and stirring was continued for 30 min. The reaction mixture was neutralized with glacial acetic acid and concentrated. Purification by flash column chromatography on silica gel (ethyl acetate / hexane 1: 1) gives (6-ethynyl-isoquinolin-4-yl) -acetic acid methyl ester M⁺ 225. This compound (79 mg, 0.35 mmol) was dissolved in methanol under an inert atmosphere and 10% Pd on carbon (79 mg) was added. The resulting suspension was stirred vigorously under an atmosphere of hydrogen gas. After 90 min. filtered, washed with methanol and concentrated to give (6-ethyl-isoquinolin-4-yl) -acetic acid methyl ester M⁺ 229. LiOH (12.5 mg) was added to a solution of this compound (68 mg, 0.30 mmol) in THF / methanol / water (3: 1: 1, 3.5 ml) and stirred for 20 h. in room temperature. After concentration under reduced pressure, (6-ethyl-isoquinolin-4-yl) -acetic acid lithium salt M⁺ 221.

Interim product 59

Intermediate 29 solution (0.5 g, 2.3 mmol) was suspended in 48% HBr aqueous solution (10 mL) and heated at 100 ° C for 48 h. Another 48% HBr aqueous solution (10 mL) was added and heated at 100 ° C for a further 24 h. The reaction mixture was cooled to 5 ° C for 4 h. and the resulting precipitate was filtered off. After washing with water and drying under high vacuum at 50 ° C, the hydrobromide of (6-hydroxy-isoquinolin-4-yl) -acetic acid [MH] was obtained.⁺ 204.4. This compound was suspended in DMF (2 mL), K2CO3 (0.22 g, 1.58 mmol) was added followed by ethyl iodide (0.085 mL, 1.06 mmol) and stirred at room temperature for 24 h. After concentration and purification by flash column chromatography on silica gel (eluent: CH2Cl2 / methanol 10: 1), ethyl ester of (6-ethoxy-isoquinolin-4-yl) -acetic acid [MH] was obtained.⁺ 260. This compound (25 mg, 0.11 mmol) was dissolved in water (1 mL), LiOH (5 mg, 0.11 mmol) was added and stirred at room temperature for 30 min. After acidification with a minimum of 6N HCl and concentration, crude (6-ethoxy-isoquinolin-4-yl) acetic acid was obtained.

Examples 1-70

Compounds of formula I which also have the formula

<img file="PL212407B1_D0021.tif" />

where r¹ to r⁴ and r⁸ to r¹³ as defined above, in free or salt form, and methods for their preparation are given in the table, while a description of the methods follows. R³ is H in all examples except No. 44 where it is a CH3 group. R⁴ is H in all examples except Nos. 25-27 and 41-43, where is CH3, R⁹ is H in all examples except No. 29 where it is a CH3 group. R¹⁰ represents H in all examples except No. 57 where it is Br and No. 75 where it is Cl. R¹³ represents H in all examples except No. 56 where it is F and No. 65 where it is Br.

PL 212 407 B1

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<td></td><td>at</td><td></td><td>at</td><td></td><td>at</td><td></td><td>at</td><td></td><td>at</td><td>at</td>

<td>Ό</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>m</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>r * rf</td><td></td><td>rH</td><td></td><td>ABOUT!</td><td></td><td>ro</td><td></td><td></td><td></td><td>m</td><td>What</td>

<td>0 = ~ l</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>loam</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>CU</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

PL 212 407 B1

<td>What ΓΡ + ι — 1st rd</td><td> 11 + 39</td><td> 11 + 40</td><td> 11 + 49</td><td>σ&gt; y + OJ</td><td>σ₍+ 0Ί H.</td><td>0Ί 'd' + FROM ! ~ ł</td><td> 14+49</td><td> 15 + 49</td>

<td></td><td></td><td> &lt;</td><td>AT</td><td> &lt;</td><td>AT</td><td>AT</td><td>AT</td><td>AT</td>

<td></td><td></td><td></td><td></td><td>ro i - 1 tp</td><td></td><td></td><td></td><td></td>

<td>+ s γ- γο +</td><td>about 00 Ή⁷</td><td>+ £ LP vo Ή *</td><td>sT &lt;M</td><td></td><td></td><td></td><td></td><td></td>

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PL 212 407 B1

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PL 212 407 B1

<td> 11 + 24</td><td>m OJ + rH rH</td><td>rH + i — 1 rH</td><td> 11 + 42 1-</td><td>r * i M.¹+ 1 — I H.</td><td>σι sf + l&gt;</td><td>ΟΊ + WHAT</td><td>σ&gt; + (Tt</td>

<td>AT</td><td>ABOUT</td><td>ABOUT</td><td>Q</td><td>Q</td><td>Q</td><td>ABOUT</td><td>Q</td>

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PL 212 407 B1

<td>σι -τ + ο Γ- (</td><td>σ \ + σ&gt; γΗ</td><td> 11 + 23</td><td>'• Ό ΓΗ + AND^-1 γ—)</td><td> 11 + 44</td><td>Γ- ΓΗ + γΗ γΗ</td><td> 11 + 28</td><td> 11 + 22</td>

<td>Ρ</td><td>Ρ</td><td>Ρ</td><td>Ο</td><td>CJ</td><td>Ρ</td><td>Ρ</td><td>AT</td>

<td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

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<td></td><td></td><td></td><td></td><td></td><td></td><td>/ \ σ)</td><td></td>

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<td></td><td>\ II</td><td>CI</td><td>&lt;Ν</td><td>CN</td><td></td><td></td><td></td>

<td>&lt;ν -γ ^</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

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<td></td><td></td><td>ο</td><td>ο</td><td>υ</td><td>υ</td><td>ο</td><td>υ</td>

<td></td><td>ΞΕ</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td>Ο</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

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<td>m</td><td>ΓΟ</td><td>γο</td><td>θ '!</td><td>m</td><td>η</td><td>m</td><td></td>

PL 212 407 B1

<td> 11 + 45</td><td>LD + r- ( H.</td><td> 11 + 47</td><td> 1</td><td>CTi OJ + i — 1 i — 1</td><td>σ&gt; OJ + &lt;o</td><td> 15 + 30</td><td>about WHAT + H [-1</td><td> 14 + 29</td><td> 14 + 30</td><td colspan="2">0Ί -sjl + tr</td>

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PL 212 407 B1

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PL 212 407 B1

<img file="PL212407B1_D0022.tif" />

PL 212 407 B1

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PL 212 407 B1

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PL 212 407 B1

<img file="PL212407B1_D0023.tif" />

PL 212 407 B1

Method A

To 5,6-diamino-1-isobutyl-3-methyl-1H-pyrimidine-2,4-dione (0.223 g, 1.05 mmol) and (6.7-dimethoxy-1-methyl-isoquinolin-4- 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (0.201 g, 1.30 mmol) was added thereto (0.25 g, 0.96 mmol) dissolved in 5 ml of methanol and 1 ml of water. . at ambient temperature. The methanol was evaporated and the obtained precipitate was filtered off, dissolved in 5 ml of methanol and 0.5 ml of 5M aqueous sodium hydroxide solution was added, heated for 1 hour. at reflux, cooled to room temperature and evaporated. The residue was dissolved in water and extracted with methylene chloride. The combined organic extracts were dried over sodium sulfate. methylene chloride was evaporated to give 8- (6,7-dimethoxy-1-methyl-isoquinolin-4-ylmethyl) -3-isobutyl-1-methyl-3,7-dihydro-purine-2,6-dione, M⁺ 437.

Method B1

(6-ethynyl-isoquinolin-4-yl) -acetic acid (58 mg, 0.28 mmol) was dissolved in 1 mL of DMF and O- (7-azabenzotriazo-1-yl) -N, N, N 'hexafluorophosphate was added. N'-tetramethyluronium (0.125 g, 0.33 mmol) and Hunig's base (0.180 mL, 1.03 mmol) followed by 5,6-diamino-1-isobutyl-3-methyl-1H-pyrimidine-2,4-dione (58 mg, 0.28 mmol) in 0.7 mL of DMF. The reaction was stirred for 2 h. in room temperature. The solvent was evaporated and the residue was purified by tlash chromatography (eluent: methylene chloride - methanol 30: 1). The purified compound was dissolved in 2 ml of methanol and 2.75 ml of water and 0.25 ml of a 4M aqueous sodium hydroxide solution were added. The reaction was heated for 2 h. at 40 ° C and then stirred for 16 hours. in room temperature. The solvent was evaporated and the crude product was purified by flash chromatography (eluent: methylene chloride - methanol 30: 1) to give 8- (6-ethynyl-isoquinolin-4-ylmethyl) -3-isobutyl-1-methyl-3,7-dihydro. purine-2,6-dione [MH]⁺ 388.

Method B2

To a suspension of (6-methoxy-isoquinolin-4-yl) -acetic acid (3.5 g, 13.82 mmol) in 70 ml of acetonitrile at room temperature, Hunig's base (6.15 ml, 36 mmol), hexafluorophosphate O - (7-benzotriazo-1-yl) -N, N, N ', N'-tetramethyluronium (6.29 g, 16.6 mmol) and 5.6-diamino-1-isobutyl-3-methyl-1H-pyrimidine- 2,4-dione (3.22 g, 15.5 mmol). The reaction was stirred for 2 hours. at ambient temperature and then the solvent is evaporated off. The residue was triturated with ethyl acetate (50 ml), filtered off, washed with ethyl acetate and dried at 50 ° C in vacuo. The dried compound was suspended in a mixture of 30 ml of methanol and 60 ml of 4M aqueous sodium hydroxide solution and heated for 45 min at 80 ° C. The slurry was neutralized with acetic acid and cooled overnight at 0-5 ° C. The precipitate formed was filtered off and washed with 30 ml of a methanol-water mixture (1: 9) and then with 30 ml of methanol. After drying at 50 ° C under high vacuum, 3-isobutyl-8- (6-methoxy-isoquinolin-4-ylmethyl) -1-methyl-3,7-dihydro-purine-2,6-dione [MH] was obtained.⁺ 394,5.

Method C

For the suspension of 5,6-diamino-1-isobutyl-3-methyl-1H-pyrimidine-2,4-dione (0.327 g, 1.54 mmol), (1-chloro-6,7-dimethoxy-isoquinolin-4 acid) -yl) -acetic acid (0.414 g, 1.54 mmol) and 1-hydroxybenzotriazole (0.251 g, 1.85 mmol) in CH2Cl2 (2 ml) were added 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (5.6M) aqueous solution, 0.33 mL, 1.85 mmol). Then ml of water was added, the two-phase mixture was shaken 18 h. and the resulting precipitate was filtered off, which was suspended in 10 ml of methanol, added 5 ml of 4M NaOH and heated for 4 hours. at boiling point. After evaporation of methanol, the residue was acidified with concentrated hydrochloric acid to pH 2, the precipitate formed was filtered off and purified using preparative HPLC to give 8- (1-chloro-6,7-dimethoxy-isoquinolin-4-ylmethyl) -3-isobutyl-1-, hydrochloride salt. methyl-3,7-dihydro-purine-2,6-dione [MH]⁺ 458.

Method D

To a mixture of 5,6-diamino-1-isobutyl-3-methyl-1H-pyrimidine-2,4-dione (20 g, 0.094 mol), 5,6-dimethoxy-isoquinolin-4-yl) -acetic acid (26 , 7 g, 0.094 mol) and 1-hydroxybenzotriazole (19.2 g, 0.142 mol) in 400 ml of methylene chloride and water mixed 1: 1 was added 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide (20.6 ml , 0.11 mole). The mixture was stirred for 4.5 hours. at ambient temperature and the resulting precipitate was filtered off, which was suspended in water (500 ml), filtered, washed with 250 ml of water and dried. After trituration with methanol and drying, a pure product was obtained, as well as a less pure product from the concentration of the methanol washings. The obtained compound (16.08 g) was dissolved in 100 ml of water and 100 ml of methanol, 56 ml of 4M aqueous sodium hydroxide solution was added and heated overnight at 70 ° C. After cooling to ambient temperature, methanol was evaporated and the residue was acidified to pH 1 with concentrated hydrochloric acid. The hydrochloride formed was filtered off and dried. The hydrochloride was converted to free

Base by adding aqueous sodium hydroxide solution to pH 11, washed with water to give 3-isobutyl-8- (5,6-dimethoxy-isoquinolin-4-ylmethyl) -1-methyl-3,7-dihydro-purine- 2,6-dione [MH]⁺ 424,6.

Method E

A suspension of the product of example 11 (72 mg, 0.13 mmol) in 6N HCl (2.5 ml) and 1.5 ml of ethanol was heated to reflux for 5 h. and then left overnight at room temperature. The resulting precipitate was filtered off, washed with water and dried to obtain 3- (3-aminobenzyl) -8- (6,7-dimethoxy-isoquinolin-4-ylmethyl) -1-methyl-3,7-dihydro-purine-2,6 dihydrochloride. -dion.¹H NMR (400MHz, DMSO) δ: 3.20 (s 3H), 3.95 (s 3H), 4.00 (s 3H). 4.75 (s, 2H). 5.15 (s 2H), 7.15 (m 2H), 7.20 (s 1H), 7.30 (t J 6 1H), 7.65 (s 1H), 7.95 (s 1H), 8.50 (s 1H), 9.50 (s 1H), 13.6 (br s 1H).

Method F

The product of example 58 (37 mg, 0.07 mmol) was suspended in 1.5 ml of pyridine and dimethylsulfamoyl chloride (23 ml, 0.21 mmol) was added. Heated for 22 hours. at 50 ° C and the solvent was evaporated. Triturate with water and filter the resulting precipitate which is dried to give 3- [3- (N, N-dimethylsulfamoyl) amino-benzyl] -8- (6,7-dimethoxy-isoquinolin-4-ylmethyl) -1-methyl-3, 7-dihydro-purine-2,6-dione.¹H NMR (400MHz, DMSO) δ: 2.64 (s 6H), 3.26 (s 3H), 3.86 (s 3H), 3.98 (s 3H), 4.50 (s 2H), 5 , 15 (s 2H), 6.98 (d J 6 1H), 7.08 (d J 6 1H), 7.15 (s 1H), 7.22 (t J 6 3H), 7.55 (s 1H), 7.62 (s 1H), 8.38 (s 1H), 9.15 (s 1H), 9.82 (s 1H), 13.60 (s 1H).

Method G.

The product of example 24 (100 mg, 0.25 mmol) was heated at 100 ° C in 5 ml of concentrated hydrobromic acid for 36 h. The solvent was evaporated and the obtained crude product was purified by preparative HPLC to give 8- (7-hydroxy-isoquinolin-4-ylmethyl) -3-isobutyl-1-methyl-3,7-dihydro-purine-2,6-dione [M]⁺ 379.

Method H.

The product of example 64 (41 mg, 0.09 mmol) was dissolved in 2 ml of acetic acid and the solution was added

100 μΐ bromine in acetic acid (148 mg / ml). It was stirred for 1 hour. at room temperature, the solvent was evaporated, the residue was dissolved in hot methanol, filtered and evaporated to give 8- (8-bromo-6,7-dihydroxy-isoquinolin-4-ylmethyl) -3-isobutyl-1-methyl-3,7- dihydro-purine-2,6-dione M⁺ 474.

Method I.

Suspension of the product of Example 13, 3-allyl-8- (6,7-dimethoxy-isoquinolin-4-ylmethyl) -1-methyl-3,7-dihydro-purine-2,6-dione hydrochloride (0.760 g, 187 mmol ), 9-borabicyclo [2.2.0] nonane (18.7 mL, 9.35 mmol, 0.5M THF solution), and diisopropylethylamine (0.33 mL, 1.89 mmol) in 9 mL THF was heated for 2 5 hours at boiling point. Then 6 ml of a 4M sodium hydroxide solution, 3 ml of 27.5% hydrogen peroxide were added sequentially, and the mixture was heated for 1.5 hours. at 50 ° C. After evaporation, the crude product was purified by flash chromatography (eluent: methylene chloride 19: 1) and then triturated with water to give 8- (6,7-dimethoxy-isoquinolin-4-ylmethyl) -3- (3-hydroxypropyl) -1 -methyl-3,7-dihydro-purine-2,6-dione [MH]⁺ 426.

Method J

To a solution of the product of example 10, 8- (6,7-dimethoxy-isoquinolin-4-ylmethyl) -3-isobutyl-1-methyl-3,7-dihydro-purine-2,6-dione (0.100 g, 0.100 g 24 mmol) in 2 mL of DMF were added potassium carbonate (48 mg, 0.35 mmol) and iodomethane (0.018 mL, 0.295 mmol). Stirred overnight, then purified using preparative HPCL to give 8- (6,7-dimethoxy-isoquinolin-4-ylmethyl) -3-isobutyl-1,7-dimethyl-3,7-dihydro-purine-2,6-. dione [MH]⁺ 438.

Method K

Suspension of the product of Example 68, 8- (6,7-dimethoxy-isoquinolin-4-ylmethyl) -3- (3-hydroxy-2-methyl-propyl) -1-methyl-3,7-dihydro-purine-2, 6-dione (63 mg, 0.14 mmol) and acetyl chloride (18 mL, 0.25 mmol) in 1 mL of pyridine was heated for 18 hours at 50 ° C. After evaporation and purification by tlash chromatography (eluent: methylene chloride - methanol 19: 1), 3- [8- (6,7-dimethoxyisoquinolin-4-ylmethyl) -1-methyl-2,6-dioxo-1,2-acetate was obtained , 6,7-tetrahydro-purin-3-yl] -2-methyl-propyl [MH]⁺ 482.

Method L.

Product of example 18,8- (6,7-dimethoxy-isoquinolin-4-ylmethyl) -1-methyl-3- (2-methyl-allyl) -3,7-dihydro-purine-2,6-dione (100 mg, 0.24 mmol), suspended in 30 ml of 1,2-dichloroethane. Diethylzinc (1.2 mL, 1.20 mmol, 1M hexane solution) was added followed by chloroiodomethane (0.174 mL, 0.24 mmol) and stirred at ambient temperature for 1 h. then a saturated aqueous NH4Cl solution was added. Extracted with chloroform, the organic layer was washed with water, dried over MgSO4 and the solvent was evaporated. Purification by preparative HPLC gave 8- (6,7-dimethoxy-isoquinolin-4-yI-methyl) -1-methyl-3- (1-methyl-cyclopropylmethyl) -3,9-dihydro-purine-2,6-dione [ ΜΗ]⁺ 436.

PL 212 407 B1

Method M

The product of example 53.8- (6-bromo-isoquinolin-4-ylmethyl) -3-isobutyl-1-methyl-3,7-dihydropurine-2,6-dione (245 mg, 0.554 mmol) was suspended in a mixture of triethylamine ( 0.085 mL, 0.61 mmol) and 4 mL CH2Cl2. While stirring, a solution of di-tert-butyl pyrocarbonate (133 mg, 0.61 mmol) in 1 mL CH2Cl2 was added dropwise; after 2 hours, triethylamine (0.170 ml, 1.2 mmol) and di-tert-butyl pyrocarbonate (130 mg, 0.60 mmol) and 0.3 ml of DMF were added again and stirred at room temperature for 2.5 days. It was then concentrated, partitioned between water and hexane, immersed in an ultrasonic bath, filtered, re-concentrated and purified by flash chromatography (on silica gel, eluent: CH2Cl2 - methanol 19: 1) to give 8- (6-bromoisoquinoline acid tert-butyl ester) -4-ylmethyl) -3-isobutyl-1-methyl-2,6-dioxo-1,2,3,6-tetrahydro-purine-7-carboxylic acid ([MH]⁺ 543). This compound (58 mg, 0.11 mmol) was added to Zn (CN) 2 (15 mg, 0.13 mmol), followed by the addition of 1,1'-bis (diphenylphosphino) -ferrocene (9 mg), tris (dibenzylideneacetone) and ) dipalladium (0) (5 mg) and anhydrous DMF (2.5 ml) and stirred for 18 hours. at a temperature of 120 ° C, and for a further 24 hours. at 150 ° C. Zn (CN) 2 (57 mg, 0.49 mmol) and 1 ml of anhydrous DMF were added and the mixture was heated for 2 h. at a temperature of 155 ° C and then 18 hours. at 145 ° C. At the end of this time, 1,1'-bis (diphenylphosphino) -ferrocene (9 mg), tris (dibenzylidene-acetone) dipalladium (0) (9 mg) were added and heating was continued for a further 6 h. at 145 ° C. Concentrated, triturated with water. filtered, washed with a 1: 1 mixture of saturated NaHCO3 and water, extracted with CH2Cl2 and with a mixture of methanol - CH2Cl2 1: 1. Purified by flash chromatography on silica gel twice (eluents: 10: 1 CH2Cl2 - methanol, then 20: 1 CH2Cl2 - methanol) to give 4- (3-iso-butyl-1-methyl-2,6-dioxo-2,3, 6,7-tetrahydro-1H-purine-8-ylmethyl) -isoquinolin-6-carbonitrile [MH]⁺ 389.

Method N

To 5,6-diamino-1-isobutyl-3-methyl-1H-pyrimidine-2,4-dione (0.40 g, 1.9 mmol) and (1-chloro-6,7-dimethoxy-isoquinolin- 1- (3-Dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (0.29 g, 1.9 mmol) was added to 4-yl) acetic acid (0.39 g, 1.78 mmol) in methanol and stirred at ambient temperature for 2 hours The methanol is evaporated off, the precipitate formed is filtered off and crystallized from an ethyl acetate-methanol mixture. The obtained product was heated in a sealed tube (100 ° C, 8h) with 40% aqueous dimethylamine solution. The mixture was concentrated and extracted with ethyl acetate. The extracts were washed with water, brine, dried over sodium sulfate, filtered, drying agent was concentrated. Purification by flash chromatography on silica gel (eluent: ethyl acetate methanol) gave 8- (1-dimethylamino-6,7-dimethoxy-isoquinolin-4-ylmethyl) -3-isobutyl-1-methyl-3,7- dihydro-purine-2,6-dione MH⁺ 467.

Method O

To 5,6-diamino-1-isobutyl-3-methyl-1H-pyrimidine-2,4-dione (0.40 g, 1.9 mmol) and (1-chloro-6,7-dimethoxy-isoquinolin- acid) 1- (3-Dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (0.29 g, 1.9 mmol) was added to 4-yl) acetic acid (0.39 g, 1.78 mmol) in methanol and stirred at ambient temperature for 2 hours The methanol is evaporated off, the resulting precipitate is filtered off, which is recrystallized from an ethyl acetate-methanol mixture. The crystallized compound was heated with piperidine for 8 h. at boiling point. The solution was filtered, washed with water and brine, dried over sodium sulfate, filtered and concentrated. Further purification was carried out by flash chromatography on silica gel (eluent: ethyl acetate - methanol). The resulting precipitate was dissolved in 20% 1N NaOH / methanol and heated for 2 h. at boiling point. Concentrate, add water and extract with ethyl acetate. The organic layers were washed with water and brine, dried over Na2SO4. the drying agent was filtered off, the solvent was evaporated to give 8- (6,7-dimethoxy-1-piperidin-1-yl-isoquinolin-4-ylmethyl) -3-isobutyl-1-methyl-3,7-dihydro-purine-2,6 - MH dione⁺ 507.

NMR for examples (¹H 400MHz DMSO-d6)

Example 12 δ 3.25 (s 3Η), 3.92 (s 3H), 4.02 (s 3H), 4.65 (s 2H), 7.70 (s 1H), 7.88 ( s 1H), 8.45 (s 1H), 9.42 (s 1H), 11.1 (s 1H), 13.60 (s 1H)

Example 13 δ 3.20 (s 3H), 4.95 (s 3H), 4.00 (s 3H), 4.52 (d J 4 2H), 4.70 (s 2H), 5, 04 (d, J 18 1H), 5.09 (d, J 10 1H), 5.88 (m 1H), 7.60 (s 1H), 7.88 (s 1H), 8.46 (s 1H ), 9.42 (s 1H), 13.7 (s 1H).

Example 14 δ 0.20-0.40 (m 4H), 1.10-1.30 (m 1H), 3.21 (s 3H), 3.81 (m 2H), 3.98 ( s 3H), 4.03 (s 3H), 4.66 (s 2H), 7.65 (s 1H), 7.85 (s 1H), 8.45 (s 1H), 9.39 (s 1H), 13.70 (s, 1H).

PL 212 407 B1

Example 15 δ 0.82 (s 9H), 3.20 (s 3H), 3.78 (s 2H), 3.99 (s 3H), 4.04 (s 3H), 7.62 ( s 1H), 7.90 (s 1H), 8.45 (s 1H), 9.44 (s 1H), 13.60 (s 1H).

Example 16 δ 0.81 (d J 7 12H), 1.98 (m 1H), 2.12 (m 1H), 3.70 (d J 8 2H), 3.78 (d J 7 2H ), 3.99 (s 3H), 4.05 (s 3H), 4.70 (s 2H), 7.65 (s 1H), 7.90 (s 1H), 8.46 (s 1H), 9, 45 (s 1H), 13.6 (s 1H).

Example 17 δ 0.80-1.10 (m 6H), 1.40-1.60 (m 4H), 1.80 (m 1H), 3.15 (s 1H), 3.76 ( d J 8 2H), 3.91 (s 3H), 4.02 (s 3H), 4.68 (s 2H), 7.60 (s 1H), 7.88 (s 1H), 8.44 ( s 1H), 13.60 (s 1H).

Example 18 δ 1.69 (s 3H), 3.21 (s 3H), 3.98 (s 3H), 4.01 (s 3H), 4.46 (s 2H), 4.52 ( s 1H), 4.68 (s 2H), 4.76 (s 1H), 7.58 (s 1H), 7.84 (s 1H), 8.45 (s 1H), 9.42 (s 1H ). 13.60 (s, 1H).

Example 19 δ 1.50-1.85 (m 4H), 3.18 (s 3H), 3.50-3.85 (m 4H), 3.95 (s 3H), 4.02 ( s 3H), 4.10-4.20 (m 1H),

4.70 (s 2H), 7.75 (s 1 H), 7.920 (s 1 H), 8.50 (s 1 H), 9.50 (s 1 H), 13.60 (br s 1 H).

Example 20 δ 0.70-0.80 (m 6H), 0.99-1.10 (m 1H), 1.20-1.25 (m 1H), 1.88-2.00 ( m 1H), 3.21 (s 3H), 3.643.80 (s 2H), 3.95 (s 3H), 4.00 (s 3H), 4.68 (s 2H), 7.60 (s 1H ), 7.80 (s 1H), 8.45 (s 1H), 9.42 (s 1H), 13.60 (br s 1H).

Example 21 δ 0.83 (t J 8 3H), 1.63 (sextet J 8 2H), 3.83 (t J 8 2H), 3.99 (s 3H), 4.05 (s 3H ), 4.69 (s 2H), 7.64 (s 1H), 7.88 (s 1H), 8.44 (s 1H), 9.42 (s 1H), 11.10 (s 1H), 13. 60 (s, 1H).

Example 23 δ 0.80 (d J 7 6H), 3.18 (s 3H), 3.75 (d J 8 2H), 4.60 (s 2H), 6.32 (s 2H), 7.71 (s 1H), 7.82 (s 1H), 8.50 (s 1H), 9.42 (s 1H), 13.50 (s 1H).

Example 49 δ 0.12-0.25 (m 4H), 1.02-1.10 (m 1H), 3.20 (s 3H), 3.68 (d, J 7 2H), 4 .00 (s 3H), 4.80 (s 2H),

7.70 (d J 9 1H), 8.21 (d, J 9 1H), 8.38 (s 1H), 9.20 (s 1H), 13.10 (s 1H).

Example 86

3-isobutyl-1-methyl-8- [1- (6-methyl-5-oxo-5,6-dihydro- [1,3] dioxolo [4,5-g] isoquinolin-8-yl) ethyl- 3,7-dihydro-purine-2,6-dione

Benzo [1,3] dioxole-5-ylmethyl- (2,2-dimethoxy-ethyl) -amine (Tetrahedron 1968, 24, 1467) was reacted with pyruvic acid according to the method of Intermediate 22 to give the hydrochloride of the acid 2- [1 , 3] dioxolo [4,5-g] isoquinolin-8-yl-propionic acid, mp 224-226 ° C. The reaction with gaseous HCl in ethanol produces the hydrochloride of the acid ethyl ester, mp 223-225 ° C. This compound (2.73 g, 10 mmol) was dissolved in 20 ml of benzene and dimethyl sulfate (1.26 g, 10 mmol) was added, stirred for 5 hours. at room temperature and then the solvent was evaporated. The crude oil was dissolved in 20 mL of water, cooled to 0-5 ° C and a solution of K3Fe (CN) 6 (5.72 g, 17.4 mmol) in 25 mL of water was added followed by sodium hydroxide (2.04 g, 51 mmol) in water (15 mL). The reaction was carried out for 1.5 hours. at 5 ° C, then concentrated hydrochloric acid was added to adjust the pH of the solution to 2. The precipitated reaction product was filtered off and crystallized from a methanol-methylene chloride mixture to give 2- (6-methyl-5-oxo-5,6-dihydro-acid). [1,3] dioxolo [4,5-g] isoquinolin-8-yl) propionic mp 290 ° C (with decomposition). This compound was converted to xanthine according to the general recipe of Method D, [MH]⁺ 452.

Example 87

8- (6,7-dimethoxy-quinoline-4-ylmethyl) -3-isobutyl-1-methyl-3,7-dihydro-purine-2,6-dione

To 10 ml of THF cooled to -70 ° C was added lithium diisopropylamide (2M pentane solution 2.46 ml, 4.92 mmol) and potassium tert-butoxide (0.552 g, 4.92 mmol) followed by 6.7-dimethoxy. -4-methyl-quinoline [J. Org. Chem., 1997, 623, 568] (1.0 g, 4.92 mmol). After 1 hour

the reaction mixture was poured into a vessel containing an excess of crushed dry ice and brought to room temperature overnight. Then, pyridine hydrochloride (0.57 g, 4.92 mmol) was added and the reaction mixture was partitioned between water and ether. The aqueous layer was evaporated, dissolved in hot methanol, activated charcoal added, filtered through Celite, evaporated to give (6,7-dimethoxy-quinolin-4-yl) -acetic acid, MH⁺ 248. This compound was converted to xanthine according to the general recipe of Method C, mp&gt; 250 ° C.