Pteridine compounds for the treatment of psoriasis

ES 2 206 402 T3

DESCRIPTION

Pteridine compounds for the treatment of psoriasis.

The present invention relates to certain heterocyclic compounds, to processes and intermediates used for their preparation, to pharmaceutical compositions containing them, and to their use in therapy.

Chemokines play an important role in immune and inflammatory responses in various diseases and disorders, including asthma and allergic diseases, as well as autoimmune conditions such as rheumatoid arthritis and atherosclerosis. These small secreted molecules are a growing superfamily of 8-14 kDa proteins characterized by a conserved cysteine ​​motif. Currently, the chemokine superfamily comprises three groups that display characteristic structural motifs, the CXC, CC and C-X3 -C families. The CXC and CC families have sequence similarity and are distinguished from each other based on a single amino acid insertion between the NH-proximal pair of cysteine ​​residues. The C-X3 -C family is distinguished from the other two families on the basis that it has a three amino acid insertion between the NH-proximal pair of cysteine ​​residues.

CXC chemokines include several potent neutrophil chemoattractants and activators, such as interleukin-8 (IL-8) and neutrophil activating peptide 2 (NAP-2).

CC chemokines include potent chemoattractants of monocytes and lymphocytes, but not neutrophils. Examples include human monocyte chemotactic proteins 1-3 (MCP-1, MCP-2, and MCP-3), RANTES (expressed and secreted by normal T cells, regulated with activation), eotaxin, and the inflammatory proteins 1α and 1β of macrophages (MIP-Ια and MIP-Ιβ).

Chemokine C-X3-C (also known as fractalkine) is a potent chemoattractant and activator of microglia in the Central Nervous System (CNS) as well as monocytes, T cells, NK cells and mast cells.

Studies have shown that the actions of chemokines are mediated by subfamilies of G protein-coupled receptors, among which are the receptors called CCR1, CCR2, CCR2A, CCR2B, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CCR10 and CCR11 (for the CC family); CXCR1, CXCR2, CXCR3, CXCR4 and CXCR5 (for the CXC family); and CX3CR 1 for the C-X3-C family. These receptors represent good targets for drug development, since agents that modulate these receptors will be useful in treating disorders and diseases such as those mentioned above.

International Patent Application WO 00/39129 describes certain pteridine derivatives, which are useful for the treatment and / or prevention of transplant rejections and / or for the treatment of inflammatory conditions.

International Patent Application WO 01/19825, which has a publication date of March 22, 2001, describes 8H-pteridin-7-ones, tetrahydropteridin-7-ones, 5H, 8H-pteridin-6,7-diones and pteridine-7-ureas that inhibit cyclin-dependent serine / threonine kinase enzymes, Wee 1 tyrosine kinase, and growth factor-mediated tyrosine kinase, and as such are useful in treating diseases and disorders of cell proliferation .

Chem. Ber., Volume 107, 1974, Manfred OH et al., Pages 339-361; Nucl. Acid. Chem, volume 2, 1978, Manfred OH et al., Pages 735-739; and Dev. Biochem, volume 4, 1978, Kiriasis et al. pages 49-53, describe 4-amino-7,8-dihydro-pteridin-7-one substituted at position 2 with alkylthio groups. These compounds are intermediates in the preparation of other products.

The present invention provides compounds of formula (I) and pharmaceutically acceptable salts or solvates thereof:

<img file="ES2206402T3_D0001.tif" />

in which:

A is a group of formula (a) or (b):

ES 2 206 402 T3

<img file="ES2206402T3_D0002.tif" />

R¹ represents a C3-C7 carbocyclic group, C1-C8 alkyl, C2-C6 alkenyl or C2-C6 alkynyl, the latter four groups being optionally substituted with one or more substituent groups independently selected from halogen atoms, -OR⁴, -NR⁵R⁶, -CONR⁵R⁶, -COOR⁷, -NR⁸COR⁹, -MR¹⁰, -SO2R¹⁰, -SO2NR⁵ R⁶, -NR⁸SO2R¹⁰, an aryl or heteroaryl group both of which may be optionally substituted with one or more substituents independently selected from groups of halogen, cyano, nitro, -OR atoms⁴ , -NR⁵R⁶, -CONR⁵R⁶, -COOR⁷, -NR⁸COR¹⁰, -MR¹⁰, -SO2R¹⁰, -SO2NR⁵R⁶, -NR⁸SO2R¹⁰, C1-C alkyl₆ or trifluoromethyl;

one of R² and R³ is hydrogen, and the other is C3 -C4 alkyl substituted with one or more hydroxy groups.

R⁴ represents hydrogen, a C1-C6 alkyl or phenyl group, the latter two being optionally substituted with one or more substituent groups independently selected from halogen, phenyl, -OR atoms¹¹ and -NR¹²R¹³;

R⁵ and R⁶ independently represent a hydrogen atom or a C1-C6 alkyl or phenyl group, the latter two being optionally substituted with one or more substituent groups independently selected from halogen, phenyl, -OR atoms¹⁴ and -NR¹⁵R¹⁶, -CONR¹⁵R¹⁶, -NR¹⁵ COR¹⁶, -SO2NR¹⁵R¹⁶, NR¹⁵SO2R¹⁶

R⁵ and R⁶ together with the nitrogen atom to which they are attached, they form a saturated 4- to 7-membered heterocyclic ring system optionally comprising an additional heteroatom selected from oxygen and nitrogen atoms, which ring system may be optionally substituted with one or more substituent groups independently selected from phenyl, -OR¹⁴, -COOR¹⁴, -NR¹⁵R¹⁶ , -CONR¹⁵R¹⁶, -NR¹⁵COR¹⁶, -SO₂NR¹⁵R¹⁶, NR¹⁵ SO2 R¹⁶ or C1-C6 alkyl, itself optionally substituted with one or more substituents independently selected from halogen atoms and -NR groups¹⁵ R¹⁶ and -OR¹⁷;

R¹⁰ represents a C1-C6 alkyl or phenyl group, each of which may be optionally substituted with one or more substituent groups independently selected from halogen, phenyl, -OR atoms¹⁷ and -NR¹⁵ R¹⁶;

X is O, S or NR⁸ ;

And it's CR¹⁸ R¹⁹ ;

Z is CR²⁰ in which R²⁰ represents a C1-C6 alkyl or phenyl group, each of which may be optionally substituted with one or more substituent groups independently selected from halogen, phenyl, -OR atoms²¹ and -NR²² R²³ , or an acyl group selected from CO₂ R²¹ or CONR²² R²³ ; and each of R⁷, R⁸, R⁹, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²¹, R²²hydrogen atom, a C1-C6 alkyl or phenyl group.

and R ²³ independently represent a

In the context of the present specification, unless otherwise indicated, an alkyl or alkenyl group, or an alkyl or alkenyl moiety in a substituent group, may be straight or branched.

Aryl groups include phenyl and naphthyl. Heteroaryl is defined as a 5- or 6-membered aromatic ring optionally containing one or more heteroatoms selected from N, S, O. Examples include pyridine, pyrimidine, thiazole, oxazole, pyrazole, imidazole, furan.

Certain compounds of formula (I) are capable of existing in stereoisomeric forms. It will be understood that the invention encompasses all geometric and optical isomers of the compounds of formula (I), and mixtures thereof, including racemates. Tautomers and their mixtures also form an aspect of the present invention.

ES 2 206 402 T3

Suitably, the R group¹ represents a C3-C7 carbocyclic group, C1-C8 alkyl, C2-C6 alkenyl or C2 -C6 alkynyl, the latter four groups being optionally substituted with one or more substituent groups independently selected from halogen atoms, -OR⁴ , -NR⁵ R⁶ , -CONR⁵ R⁶ , -COOR⁷ , -NR⁸ COR⁹ , -MR¹⁰ , -SO2 R¹⁰ , -SO2 NR⁵ R⁶ , -NR⁸ SO2 R¹⁰ , an aryl or heteroaryl group both of which may be optionally substituted with one or more substituents independently selected from groups of halogen, cyano, nitro, atoms,

-OR⁴, -NR⁵R⁶, -CONR⁵R⁶, -COOR⁷, -NR⁸COR¹⁰, -MR¹⁰, -SO2 R¹⁰, -SO2NR⁵R⁶, -NR⁸SO2R¹⁰, C1-C6 alkyl or trifluoromethyl. The substituents can be present in any suitable position of the R groups¹ , aryl and heteroaryl, including nitrogen atoms of heteroaryl groups. Particularly advantageous compounds of formula (I) are those in which R¹ represents an optionally substituted benzyl group. More preferably, R¹ represents benzyl or benzyl substituted with one or more C1-C6 alkyl, C1-C6 alkoxy, or halogen atoms, even more preferably benzyl substituted with two fluorine atoms, and most preferably benzyl substituted with two fluorine atoms that are ortho and meta to the benzyl CH2 bond.

Suitably, R² represents hydrogen or a C3 -C7 carbocyclic group, C1 -C8 alkyl, C2 -C6 alkenyl or C2 -C6 alkynyl, the latter four groups being optionally substituted with one or more substituent groups independently selected from halogen atoms, -OR⁴ , -NR⁵ R⁶ -CONR⁵ R⁶ , -COOR⁷ , -NR⁸ COR⁹ , -MR¹⁰ , -SO2 R¹⁰ , -SO2 NR⁵ R⁶ or -NR⁸ SO2 R⁹ , and R³ represents hydrogen or C2-C6 alkyl optionally substituted with one or more substituent groups independently selected from halogen, phenyl, -OR atoms¹⁰ and -NR¹¹ R¹² ; or R² and R³ represent a 3-8 membered ring optionally containing one or more atoms selected from O, S, NR⁸and itself is optionally substituted with C1-3 alkyl, halogen or OR⁴ .

Preferably one of R² and R³ is hydrogen, and the other is CH (CH3) CH2OH, CH (Et) CH2OH, C (CH3) 2CH2OH, or CH (CH2 OH) 2. When one of R² and R³ is hydrogen, and the other is CH (CH 3) CH2 OH or CH (Et) CH2 OH, the resulting compounds of formula (I) are preferably in the form of the (R) isomer. Most preferably one of R² and R³ is hydrogen, and the other is CH (CH3) CH2 OH.

When A is a group of formula (a), X represents O, S or NR⁸ . When X represents NR⁸ , then R⁸ it is preferably hydrogen or C1-6 alkyl. Preferably X is S. Suitably Y is CR¹⁸ R¹⁹, preferably Y is CH₂ . ^-

Preferably A is a group of formula (b) and Z is CR²⁰ most preferably Z is CH.

Particularly preferred compounds of the invention include:

4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -2 - [(phenylmethyl) thio] -6H-pyrimido [5,4-b] [1,4] thiazin-7 (8H) - ona

2 - [[(2,3-Difluorophenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

2 - [[(2,3-Difluorophenyl) methyl] thio] -4 - [[2-hydroxy-1- (hydroxymethyl) ethyl] amino] -7 (8H) -pteridinone

2 - [[(2,3-Difluorophenyl) methyl] thio] -4 - [(2-hydroxyethyl) amino] -7 (8H) -pteridinone (2R) -2 - [[2 - [[2,3-Difluorophenyl ) methyl] thio] -7,8-dihydro-7-oxo-4-pteridinyl] amino] -propanamide

2 - [[(3-Chloro-2-fluorophenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

2 - [[(3-Chloro-2-fluorophenyl) methyl] thio] -4 - [(2-hydroxyethyl) -amino] -7 (8H) -pteridinone

2 - [[(3-Chloro-2-fluorophenyl) methyl] thio] -4 - [[2-hydroxy-1- (hydroxymethyl) ethyl] amino] -7 (8H) -pteridinone

[(2R) -2 - [[2 - [[(3-chloro-2-fluorophenyl) methyl] thio] -7,8-dihydro-7-oxo-4-pteridinyl] amino acid 1,1-dimethylethyl ester ] propyl] -carbamic

4 - [[(1R) -2-amino-1-methylethyl] amino] -2 - [[(3-chloro-2-fluorophenyl) methyl] thio] -7 (8H) -pteridinone monohydrochloride

2 - [[(3-Chloro-4-methoxyphenyl) methyl] thio] -4 - [[2-hydroxy-1- (hydroxymethyl) ethyl] amino] -7 (8H) -pteridinone

4 - [(2-aminoethyl) amino] -2 - [[(3-chloro-4-methoxyphenyl) methyl] thio] -7 (8H) -pteridinone monotrifluoroacetate

2 - [[(2-Fluoro-4-methoxyphenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

2 - [[(2-Fluoro-3-methylphenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

2 - [[(3-Fluoro-2-methoxyphenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

2 - [[[4- (Difluoromethoxy) phenyl] methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

ES 2 206 402 T3

4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -2 - [[(4-hydroxyphenyl) -methyl] thio] -7 (8H) -pteridinone

4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -2 - [[(3-methylphenyl) -methyl] thio] -7 (8H) -pteridinone

2 - [(1,3-Benzodioxol-4-ylmethyl) thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

2 - [[(2,4-Difluorophenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

2 - [[(3-Chlorophenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -2 - [(5-isoxazolyl-methyl) thio] -7 (8H) -pteridinone

4 - [[- 2-hydroxy-1- (hydroxymethylethyl] amino] -2 - [(5-isoxazolylmethyl) thio] -7 (8H) -pteridinone

4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -2 - [[(5-methyl-2-furanyl) methyl] thio] -7 (8H) -pteridinone

2 - [[(2-Fluorophenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methyl-ethyl] amino] -7 (8H) -pteridinone

4 - [[(1R) -2-Hydroxy-1-methylethyl] amino] -2 - [[(2-thienyl) -methyl) thio] -7 (8H) -pteridinone

2 - [[(2-Fluorophenyl) methyl] thio] -4 - [[- 2-hydroxy-1- (hydroxy-methylethyl] amino] -7 (8H) -pteridinone

4 - [[- 2-Hydroxy-1- (hydroxymethyl) ethyl] amino] -2 - [(2-thienylmethyl) thio] -7 (8H) -pteridinone

2 - [[(2-Fluoro-5-methylphenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

2 - [[(2-Fluoro-3- (trifluoromethyl) phenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

2 - [[(2-Fluoro-3- (trifluoromethyl) phenyl) methyl] thio] -4 - [[2-hydroxy-1- (hydroxymethyl) ethyl] amino] -7 (8H) -pteridinone

2 - [[(2-Fluoro-3- (trifluoromethyl) phenyl) methyl] thio] -4 - [(2-hydroxyethyl) amino] -7 (8H) -pteridinone

4 - [(2-Aminoethyl) amino] -2 - [[(2-fluoro-3- (trifluoromethyl) -phenyl) methyl] thio] -7 (8H) -pteridinone

2 - [[(2,3-Difluorophenyl) methyl] thio] -4 - [[2-hydroxy-1- (hydroxymethyl) -1-methylethyl] amino] -7 (8H) -pteridinone

2 - [[(2,3-Difluorophenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -6-methyl-7 (8H) -pteridinone

2 - [[(2,3-Difluorophenyl) -methyl] thio] -7,8-dihydro-4 - [[(1R) -2-hydroxy-1-methylethyl] -amino] -7-oxo6 acid ethyl ester -pteridinecarboxylic

2 - [[(2,3-Difluorophenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -6- (trifluoromethyl) -7 (8H) -pteridinone Sodium salt of 2 - [[(2,3-Difluorophenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone 2 - [(2,3-Difluorobenzyl ) thio] -4- (ethylamino) -7 (8H) -pteridinone

2 - [(2,3-Difluorobenzyl) thio] -4- (isopropylamino) -7 (8H) -pteridinone (+/-) - 4- (sec-Butylamino) -2 - [(2,3-difluorobenzyl) thio ] -7 (8H) -pteridinone

2 - ({2 - [(2,3-Difluorobenzyl) thio] -7-oxo-7,8-dihydro-4-pteridinyl} amino) acetamide (+/-) - 2 - [(2,3-Difluorobenzyl) thio] -4 - [(2-hydroxypropyl) -amino] -7 (8H) -pteridinone (S) -2 - [(2,3-Difluorobenzyl) thio] -4 - [(2-hydroxy-1-methyl- ethyl) amino] -7 (8H) -pteridinone (+/-) - 2 - [(2,3-Difluorobenzyl) thio] -4 - [(2-hydroxy-1-methyl-ethyl) amino] -7 (8H ) -pteridinone (R) -2 - [(2,3-Difluorobenzyl) thio] -4 - [(2-hydroxypropyl) -amino] -7 (8H) -pteridinone

2 - [(2,3-Difluorobenzyl) thio] -4 - [(3-hydroxypropyl) amino] -7 (8H) -pteridinone

2 - [(2,3-Difluorobenzyl) thio] -4 - [(2-hydroxyethyl) (methyl) -amino] -7 (8H) -pteridinone

3 - [{2 - [(2,3-Difluorobenzyl) thio] -7-oxo-7,8-dihydro-4-pteridinyl} (methyl) amino] propanenitrile

ES 2 206 402 T3 (R) -2 - [(2,3-Difluorobenzyl) thio] -4 - {[1- (hydroxymethyl) -propyl] amino} -7 (8H) -pteridinone (R) -2- [ (2,3-Difluorobenzyl) thio] -4 - {[1- (hydroxymethyl) -propyl] amino} -7 (8H) -pteridinone

2 - [(2,3-Difluorobenzyl) thio] -4 - [(4-hydroxybutyl) amino] -7 (8H) -pteridinone (+/-) - 2 - [(2,3-Difluorobenzyl) thio] -4 - {[1- {hydroxymethyl) -propyl] amino} -7 (8H) -pteridinone

2 - [(2,3-Difluorobenzyl) thio] -4 - [(2-hydroxy-1,1-dimethyl-ethyl) amino] -7 (8H) -pteridinone

2 - [(2,3-Difluorobenzyl) thio] -4- [ethyl (2-hydroxyethyl) amino] -7 (8H) -pteridinone (+/-) - 4 - [(3-Amino-2-hydroxypropyl) amino ] -2 - [(2,3-Difluorobenzyl) thio] -7 (8H) -pteridinone (+/-) - 2 - [(2,3-Difluorobenzyl) thio] -4 - [(1,3-dimethylbutyl) -amino] -7 (8H) -pteridinone (1R, 2R) -2 - [(2,3-difluorobenzyl) thio] -4 - [(2-hydroxy-cyclopentyl) amino] -7 (8H) -pteridinone

2 - [(2,3-Difluorobenzyl) thio] -4 - [(5-hydroxypentyl) amino] -7 (8H) -pteridinone (+/-) - 2 - [(2,3-difluorobenzyl) thio] -4 - {[1- (hydroxymethyl) -butyl] amino} -7 (8H) -pteridinone (+/-) - 2 - ({2 - [(2,3-difluorobenzyl) thio] -7-oxo-7.8 Methyl-dihydro-4-pteridinyl} amino) propanoate 2 - [(2,3-difluorobenzyl) thio] -4 - [(3-hydroxy-2,2-dimethyl-propyl) amino] -7 (8H) -pteridinone (1R, 2R) -2 - [(2,3-difluorobenzyl) thio] -4 - {[2-hydroxy-1- (hydroxymethyl) propyl] amino} -7 (8H) -pteridinone

4- [bis (2-hydroxyethyl) amino] -2 - [(2,3-difluorobenzyl) thio] -7 (8H) -pteridinone

2 - [(2,3-Difluorobenzyl) thio] -4 - {[2- (2-hydroxyethoxy) ethyl] -amino} -7 (8H) -pteridinone

2 - [(2,3-Difluorobenzyl) thio] -4 - [(2,2-dimethoxyethyl) amino] -7 (8H) -pteridinone

4 - {[2- (diethylamino) ethyl] amino} -2 - [(2,3-difluorobenzyl) -thio] -7 (8H) -pteridinone (S) -2 - [(2,3-difluorobenzyl) thio] -4 - {[1- (hydroxymethyl) -2,2-dimethylpropyl] amino} -7 (8H) -pteridinone (R) -2 - [(2,3-difluorobenzyl) thio] -4 - {[1- ( hydroxymethyl) -3-methylbutyl] amino} -7 (8H) -pteridinone

2 - [(2,3-difluorobenzyl) thio] -4 - [(6-hydroxyhexyl) amino] -7 (8H) -pteridinone

2 - [(2,3-Difluorobenzyl) thio] -4 - {[3- (dimethylamino) -2,2-dimethylpropyl] amino} -7 (8H) -pteridinone (S) -2 - ({2 - [( Ethyl 2,3-difluorobenzyl) thio] -7-oxo-7,8-dihydro-4-pteridinyl} amino) -3-hydroxypropanoate and pharmaceutically acceptable salts and solvates thereof.

According to the invention, there is also provided a process for the preparation of a compound of formula (I), comprising:

(a) treating a compound of formula (IIA):

Ν Ν S — R

Η (ΠΑ) in which R¹ is as defined in formula (I), or is a protected derivative thereof, and L is a leaving group, with an amine HNR² R³, or (b) treating a compound of formula (IIB):

ES 2 206 402 T3

<img file="ES2206402T3_D0003.tif" />

in which R¹ is as defined in formula (I), or is a protected derivative thereof, and L is a leaving group, with an amine HNR² R³ , or (c) treating a compound of formula (IIC):

<img file="ES2206402T3_D0004.tif" />

in which R¹, R² and R³ are as defined in formula (I), or are protected derivatives thereof, with a thiol R¹SH, or (d) treating a compound of formula (IID):

<img file="ES2206402T3_D0005.tif" />

in which R¹ is as defined in formula (I), or is a protected derivative thereof, and L 'is a leaving group, with an amine HNR² R³ , and optionally after process (a), (b), (c) or (d), and in any order:

- remove any of the protecting groups

- form a pharmaceutically acceptable salt.

The reaction of compounds of formula (IIA) and (IIB) with an amine HNR²R³ It can be carried out in a solvent such as N-methyl-pyrrolidinone at a temperature between 0 ° C and 150 ° C. Suitable leaving groups L include halogen, especially chlorine or bromine.

The reaction of compounds (IIC) with a thiol R¹SH can be carried out in a solvent, such as N-methylpyrrolidinone, using a base, such as potassium tert-butoxide, at a temperature between 0 ° C and 150 ° C.

Reaction of compounds of formula (IID) with an amine HNR²R³ can be carried out using a pure amine HNR²R³ or in a solvent such as 1-methylimidazole at a temperature between 50 ° C and 200 ° C, with or without the aid of microwave radiation.

Compounds of formula (IIA), in which R¹ is as defined in formula (I) and L is a leaving group, such as chlorine, can be prepared by treating a compound of formula (IIA), in which R¹ is as defined above and L is a hydroxyl group, with a halogenating agent, such as phosphorous oxychloride. The reaction can be carried out under reflux in the presence of N, N-dimethylaniline.

Compounds of formula (IIA), in which R¹ is as defined in formula (I) and L is a hydroxyl group, they can be prepared by acid treatment of a compound of formula (III), in which R¹ and L are as defined above. Suitable acids include p-toluenesulfonic acid, and the reaction can be carried out in a solvent such as refluxing toluene.

ES 2 206 402 T3

<img file="ES2206402T3_D0006.tif" />

Compounds of formula (III), in which R¹ is as defined in formula (I) and L is a hydroxyl group, they can be prepared by treating a compound of formula (IV), in which R¹ and L are as defined above, with a reducing agent in the presence of ethyl bromoacetate. The reaction can be carried out in a solvent such as ethanol at room temperature using a reducing agent such as sodium borohydride.

<img file="ES2206402T3_D0007.tif" />

Compounds of formula (IV), in which R¹ is as defined in formula (I) and L is a hydroxyl group, they can be prepared by treating a compound of formula (V), in which R¹ and L are as defined above, with a metal thiocyanate in the presence of bromine. The reaction can be carried out in a solvent such as N, N-dimethylformamide, at a temperature between 0 ° C and 100 ° C in the presence of pyridine, using potassium thiocyanate.

<img file="ES2206402T3_D0008.tif" />

Compounds of formula (V), in which R¹ is as defined in formula (I) and L is a hydroxyl group, they are suitably prepared by reacting a compound of formula (VI):

<img file="ES2206402T3_D0009.tif" />

with a compound of formula R¹X where R¹ is as defined above and X is a leaving group, such as bromide, in the presence of a base such as sodium hydroxide. The reaction can be carried out in aqueous NMP at room temperature.

Compounds of formula (VI) are commercially available.

Compounds of formula (IIB), in which R¹ is as defined in formula (I) and L is a leaving group, such as bromine, can be prepared by treating a compound of formula (IIB), wherein R¹ is as defined above and L is NH2, with a diazotizing agent, such as isoamyl nitrite, in the presence of a halogenating agent, such as bromoform. The reaction can be carried out in a solvent, such as DMSO, at a temperature between 0 ° C and 100 ° C.

ES 2 206 402 T3

Compounds of formula (IIB), in which R¹ is as defined in formula (I) and L is NH2, the following can be prepared: i) By treating a compound of formula (VII):

<img file="ES2206402T3_D0010.tif" />

in which R¹ and L are as defined above, with ethyl glyoxylate in the presence of a base, such as sodium methoxide, in a solvent, such as methanol, at room temperature, or ii) Treatment of a compound of formula (VIII):

<img file="ES2206402T3_D0011.tif" />

in which R¹ and L are as defined above, with triethyl phosphonoacetate in the presence of a base such as butyllithium. The reaction can be carried out in a solvent, such as DMF, at a temperature between 0 ° C and 100 ° C.

Compounds of formula (VII), in which R¹ is as defined in formula (I) and L is NH2, they can be prepared by treating a compound of formula (VIII), in which R¹ and L are as defined above, with a reducing agent, such as sodium hydrosulfite. The reaction can be carried out in a solvent, such as water, under reflux.

<img file="ES2206402T3_D0012.tif" />

Compounds of formula (VIII), in which R¹ is as defined in formula (I) and L is NH2, they can be prepared by treating a compound of formula (IX), in which R¹ and L are as defined above, with a nitrosating agent, such as sodium nitrite. The reaction can be carried out in a solvent, such as aqueous acetic acid, at a temperature between 0 ° C and 100 ° C.

<img file="ES2206402T3_D0013.tif" />

Compounds of formula (IX), in which R¹ is as defined in formula (I) and L is NH2, they can be prepared by treating a compound of formula (X) with a compound of formula R¹X, in which R¹ is as defined above and X is a leaving group, such as bromide, in the presence of a base, such as potassium tert-butoxide. The reaction can be carried out in a solvent, such as DMSO, at room temperature.

ES 2 206 402 T3

<img file="ES2206402T3_D0014.tif" />

Compounds of formula (IIC), in which R¹ , R² and R³ are as defined in formula (I), they can be prepared from compounds of formula (I), in which R¹ , R² and R³ they are as defined in formula (I), by treatment with a peracid, such as peracetic acid. The reaction can be carried out in a solvent, such as acetic acid, at a temperature between 0 ° C and 100 ° C.

Compounds of formula (IID), in which R¹ is as defined in formula (I) and L is an alkoxy group, they can be prepared from compounds of formula (IID), in which R¹ is as defined in formula (I) and L is a thioalkyl group, by treatment with a suitable alcohol in the presence of a base. Suitable alcohols include isopropanol, and the base can be sodium, and the reaction can be carried out at a temperature between 0 ° C and 150 ° C.

Compounds of formula (IID), in which R¹ is as defined in formula (I) and L is a thioalkyl group, they can be prepared from compounds of formula (XI) by sequential treatment with an alkyl halide R¹X followed by ethyl glyoxalate in the presence of a base. The reaction can be carried out in a solvent, such as methanol, at a temperature between 0 ° C and 150 ° C, using sodium as the base.

<img file="ES2206402T3_D0015.tif" />

Compounds of formula (X) and (XI) are commercially available.

It will be appreciated by those skilled in the art that, in the processes of the present invention, certain functional groups, such as hydroxyl or amino groups, in the starting reagents or intermediates may need to be protected by protecting groups. Thus, the preparation of the compounds of formula (I) may involve, in an appropriate step, the removal of one or more protecting groups. The protection and deprotection of functional groups is fully described in "Protective Groups in Organic Chemistry", edited by JWF McOmie, Plenum Press (1973), and "Protective Groups in Organic Synthesis", 2^to ed. TW Greene and PGM Wuts, Wiley-Interscience (1991).

The novel intermediates form a further aspect of the invention. In particular, the compounds of formula (IIA) and (IIB) are new, and form an aspect of the invention.

The compounds of formula (I) above can be converted into a pharmaceutically acceptable salt or solvate thereof, preferably a base addition salt, such as the sodium, potassium, calcium, aluminum, lithium, magnesium, zinc, benzathine salt. , chloroprocaine, choline, diethanolamine, ethanolamine, ethyldiamine, meglumine, tromethamine or procaine, or an acid addition salt, such as a hydrochloride, hydrobromide, phosphate, acetate, fumarate, maleate, tartrate, citrate, oxalate, methanesulfonate or p-toluenesulfonate.

The compounds of formula (I) have activity as pharmaceuticals, in particular as modulators of the activity of chemokine receptors (especially CXCR2), and can be used in the treatment (therapeutic or prophylactic) of conditions / diseases in humans and non-animals. humans that are exacerbated or caused by excessive or unregulated production of chemokines. Examples of such conditions / diseases include:

(1) (the respiratory system) obstructive airway diseases including chronic obstructive pulmonary disease (COPD); asthma, such as bronchial, allergic, intrinsic, extrinsic and dust asthma, particularly chronic or deep-seated asthma (eg, late asthma and airway hypersensitivity); bronchitis; acute, allergic, atrophic and chronic rhinitis including caseous rhinitis, hypertrophic rhinitis, purulent rhinitis, dry rhinitis and drug rhinitis; membranous rhinitis including croup, fibrinous, and pseudomembranous rhinitis and rhinitis

ES 2 206 402 T3 scrofulose; seasonal rhinitis including nervous rhinitis (hay fever) and vasomotor rhinitis; sarcoidosis, farmer's lung and related diseases, fibroid lung, and idiopathic interstitial pneumonia;

(2) (bone and joints) rheumatoid arthritis, seronegative spondyloarthropathies (including ankylosing spondylitis, psoriatic arthritis, and Reiter's disease), Behcet's disease, Sjogren's syndrome, and systemic sclerosis;

(3) (skin) psoriasis, atopic dermatitis, contact dermatitis and other eczematous dermatitis, seborrheic dermatitis, lichen planus, pemphigus, pemphigus bullosa, epidermolysis bullosa, urticaria, angiodermas, vasculitis, erythema, cutaneous eosinophilia, areataveitis, and cutaneous alounciae, areataveitis, vernal;

(4) (gastrointestinal tract) celiac disease, proctitis, eosinophilic gastroenteritis, mastocytosis, Crohn's disease, ulcerative colitis, food-related allergies that have effects remote from the intestine, eg, migraine, rhinitis, and eczema;

(5) (central and peripheral nervous system) neurodegenerative diseases and dementia disorders, for example Alzheimer's disease, amyotrophic lateral sclerosis and other motor neuronal diseases, Creutzfeldt-Jacob disease and other prion diseases, HIV encephalopathy (dementia complex of the AIDS), Huntington's disease, frontotemporal dementia, Lewy body dementia, and vascular dementia; polyneuropathies, eg, Guillain-Barré syndrome, chronic demyelinating inflammatory polyradiculoneuropathy, multifocal motor neuropathy, plexopathies; CNS demyelination, eg, multiple sclerosis, acute disseminated / hemorrhagic encephalomyelitis, and subacute sclerosing panencephalitis; neuromuscular disorders, eg, myasthenia gravis and Lambert-Eaton syndrome; spinal disorders, eg, tropical spastic paraparesis, and stiff man syndrome: paraneoplastic syndromes, eg cerebellar degeneration and encephalomyelitis; CNS trauma; migraine; and stroke.

(6) (other tissues and systemic disease) atherosclerosis, Acquired Immune Deficiency Syndrome (AIDS), lupus erythematosus, systemic lupus, erythematosus, Hashimoto's thyroiditis, type I diabetes, nephrotic syndrome; eosinophilic fasciitis, hyperIgE syndrome, lepromatous leprosy, and idiopathic thrombocytopenic purpura; postoperative adhesions, and sepsis.

(7) acute or chronic (allograft rejection) following, for example, kidney, heart, liver, lung, bone marrow, skin and cornea transplantation; and chronic host versus graft disease;

(8) Cancers, especially non-small cell lung cancer (NSCLC), malignant melanoma, prostate cancer and squamous sarcoma, and tumor metastases;

(9) Diseases in which angiogenesis is associated with elevated levels of CXCR2 chemokines (eg, NSCLC, diabetic retinopathy).

(10) Cystic fibrosis, reperfusion injury to the heart, brain, peripheral limbs, and other organs.

(11) Minor wounds and chronic skin ulcers (12) Diseases of the reproductive system (eg ovulation, menstruation and implantation disorders, pain before delivery, endometriosis).

Thus, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, as defined hereinbefore, for use in therapy.

Preferably, the compounds of the invention are used to treat diseases in which the chemokine receptor belongs to the CXC chemokine receptor subfamily; more preferably, the target chemokine receptor is the CXCR2 receptor.

The particular conditions that can be treated with the compounds of the invention are psoriasis, diseases in which angiogenesis is associated with elevated levels of CXCR2 chemokines, and COPD. It is preferred that the compounds of the invention are used to treat psoriasis.

As a further aspect of the present invention, certain compounds of formula (I) may have utility as CX3CR1 receptor antagonists. Such compounds are expected to be particularly useful in treating disorders of the central and peripheral nervous system, and other conditions characterized by activation of microglia and / or infiltration of leukocytes (eg, stroke / ischemia and brain trauma).

In a further aspect, the present invention provides the use of a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, as defined hereinbefore, in the manufacture of a medicament for use in therapy.

In still a further aspect, the present invention provides the use of a compound of formula (I), or a

ES 2 206 402 T3 pharmaceutically acceptable salt or solvate thereof, as defined hereinbefore, in the manufacture of a medicament for the treatment of human diseases or morbid conditions in which modulation of chemokine receptor activity is beneficial .

In the context of the present specification, the term "therapy" also includes "prophylaxis", unless there are specific indications to the contrary. The terms "therapeutically" and "therapeutically" should be construed accordingly.

The invention still further provides a compound for use in a method of treating chemokine-mediated disease, wherein the chemokine binds to a chemokine receptor (especially CXCR2), comprising administering to a patient a therapeutically effective amount of a compound. of formula (I), or a pharmaceutically acceptable salt or solvate thereof, as defined hereinbefore.

The invention also provides a compound for use in a method of treating an inflammatory disease, especially psoriasis, in a patient suffering from, or at risk of, said disease, which comprises administering to the patient a therapeutically effective amount of a compound of formula ( I), or a pharmaceutically acceptable salt or solvate thereof, as defined hereinbefore.

For the aforementioned therapeutic uses, the dose administered will, of course, vary with the compound employed, the mode of administration, the treatment desired, and the disorder diagnosed.

The present invention also provides a compound of formula (IA) for use in a method of treating chemokine-mediated disease, wherein the chemokine binds to one or more chemokine receptors, comprising administering to a patient a therapeutically effective amount of a compound of formula (IA), or a pharmaceutically acceptable salt or solvate thereof

<img file="ES2206402T3_D0016.tif" />

in which:

A is a group of formula (a) or (b):

<img file="ES2206402T3_D0017.tif" />

Η

Φ)

R¹ represents a C3 -C7 carbocyclic group, C1-C8 alkyl, C2 -C6 alkenyl or C2 -C6 alkynyl, the latter four groups being optionally substituted with one or more substituent groups independently selected from halogen atoms, -OR⁴, -NR⁵R⁶, -CONR⁵R⁶, -COOR⁷, -NR⁸ COR⁹, -MR¹⁰, -SO2R¹⁰, -SO2NR⁵R⁶, -NR⁸SO2R¹⁰, an aryl or heteroaryl group both of which may be optionally substituted with one or more substituents independently selected from groups of halogen, cyano, nitro, -OR atoms⁴, -NR⁵R⁶, -CONR⁵R⁶ , -COOR⁷, -NR⁸COR¹⁰ , -MR¹⁰, -SO2R¹⁰, -SO2NR⁵R⁶, -NR⁸SO2R¹⁰, C1-C6 alkyl or trifluoromethyl;

R² represents hydrogen or a C3 -C7 carbocyclic group, C1 -C8 alkyl, C2 -C6 alkenyl or C2 -C6 alkynyl, being

ES 2 206 402 T3 these last four groups optionally substituted with one or more substituent groups independently selected from halogen atoms, -OR⁴, -NR⁵R⁶, -CONR⁵R⁶, -COOR⁷, -NR⁸COR⁹, -MR¹⁰, -SO2R¹⁰, -SO2NR⁵R⁶,

-NR⁸SW₂R⁹;

R³ represents hydrogen or C2-C6 alkyl optionally substituted with one or more substituent groups independently selected from halogen, phenyl, -OR atoms¹⁰ and -NR¹¹ R¹² ;

R² and R³ represent a 3-8 membered ring optionally containing one or more atoms selected from O, S, NR⁸ and itself optionally substituted with C1-3 alkyl, halogen or OR⁴;

R⁴ represents hydrogen, a C1-C6 alkyl or phenyl group, the latter two being optionally substituted with one or more substituent groups independently selected from halogen, phenyl, -OR atoms¹¹ Y

-NR¹²R¹³;

R⁵ and R⁶ independently represent a hydrogen atom or a C1-C6 alkyl or phenyl group, the latter two being optionally substituted with one or more substituent groups independently selected from halogen, phenyl, -OR atoms¹⁴ and -NR¹⁵R¹⁶, -CONR¹⁵R¹⁶, -NR¹⁵COR¹⁶, -SO2NR¹⁵R¹⁶, NR¹⁵SO2R¹⁶

R⁵ and R⁶, together with the nitrogen atom to which they are attached, form a saturated 4- to 7-membered heterocyclic ring system optionally comprising an additional heteroatom selected from oxygen and nitrogen atoms, which ring system may be optionally substituted with one or more substituent groups independently selected from phenyl, -OR¹⁴, -COOR¹⁴, -NR¹⁵R¹⁶ , -CONR¹⁵R¹⁶, -NR¹⁵COR¹⁶, -SO₂NR¹⁵R¹⁶, NR¹⁵SO2R¹⁶ or C1-C6 alkyl, itself optionally substituted with one or more substituents independently selected from halogen atoms and -NR groups¹⁵ R¹⁶ and -OR¹⁷;

R¹⁰ represents a C1-C6 alkyl or phenyl group, each of which may be optionally substituted with one or more substituent groups independently selected from halogen, phenyl, -OR atoms¹⁷ and -NR¹⁵R¹⁶;

X is O, S or NR⁸ ;

And it's CR¹⁸ R¹⁹ ;

Z is CR²⁰ in which R²⁰ represents a C1-C6 alkyl or phenyl group, each of which may be optionally substituted with one or more substituent groups independently selected from halogen, phenyl, -OR atoms²¹ and -NR²²R²³, or an acyl group selected from CO₂R²¹ or CONR²²R²³; and each of R⁷, R⁸, R⁹, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹ , R²¹, R²² hydrogen atom, a C1-C6 alkyl or phenyl group.

and R²³ independently represent a

The invention further provides a compound of formula (IA) in the above method wherein the chemokine receptor belongs to the CXC chemokine receptor family, or more specifically is the CXCR2 receptor.

The compounds of formula (I), and the pharmaceutically acceptable salts and solvates thereof, can be used by themselves, but will generally be administered in the form of a pharmaceutical composition in which the compound of formula (I) / salt / solvate (active ingredient) is in association with a pharmaceutically acceptable adjuvant, diluent or carrier. Depending on the mode of administration, the pharmaceutical composition will preferably comprise from 0.05 to 99% by weight (percent by weight), more preferably from 0.05 to 80% by weight, even more preferably from 0.10 to 70% by weight, and even more preferably 0.10 to 50% by weight, of active ingredient, all percentages by weight being based on the total composition.

The present invention also provides a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, as defined hereinbefore, in association with a pharmaceutically acceptable adjuvant, diluent or carrier.

The invention further provides a process for the preparation of a pharmaceutical composition of the invention, which comprises mixing a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, as defined hereinbefore, with an adjuvant, diluent or pharmaceutically acceptable carrier.

The pharmaceutical compositions can be administered topically (eg, to the lung and / or respiratory tract, or to the skin) in the form of solutions, suspensions, heptafluoroalkan aerosols, and dry powder formulations; or systemically, for example, by oral administration in the form of tablets, capsules, syrups, powders or granules, or by parenteral administration in the form of solutions or suspensions, or by subcutaneous administration, or by rectal administration in the form of suppositories, or transdermally. Preferably, the compounds of the invention are administered orally.

ES 2 206 402 T3

The invention will now be further illustrated with reference to the following Examples. In the examples, Nuclear Magnetic Resonance (NMR) spectra were measured on a Varian Unity Inova 300 or 400 MHz spectrometer, and Mass Spectrometry (MS) spectra were measured on a Finnigan Mat SSQ7000 or Micromass Platform spectrometer. When necessary, reactions were carried out under an inert nitrogen or argon atmosphere. Chromatography was generally performed using Matrex Silica 60^® (35-70 microns) or Prolabo Silica gel 60^® (35-70 microns) suitable for flash chromatography on silica gel. Purification by high pressure liquid chromatography was performed using a Waters Micromass LCZ with a Waters 600 pump controller, a Waters 2487 detector and a Gilson FC024 fraction collector, or a Waters Delta Prep 4000. Abbreviations pf and DMSO used in examples represent melting point and dimethylsulfoxide, respectively. Example 1

4 - [[(1R) -2-hydroxy-1-methyl-ethyl] amino] -2 - [(phenylmethyl) thio] -6H-pyrimido [5,4-bJ [1,4] thiazin- 7 (8H) -one (a) 6-amino-1,4-dihydro-2 - [(phenylmethyl) thio] -4-oxo-5-thiocyanic acid pyrimidinyl ester

6-amino-2 - [(phenylmethyl) thio] -4 (1H) -pyrimidinone (10.5 g) [preparation as described in WO 9635678] and potassium thiocyanate (25 g ) in N, N-dimethylformamide (200 ml). Pyridine (6.3 ml) was added, and the solution was cooled to 5 ° C. Bromine (2.2 ml) was added slowly, and the reaction mixture was stirred for 2 hours at 5-10 ° C. The reaction mixture was poured into ice water, stirred for 1 hour, and the solid was isolated by filtration. After washing with water and ether, a pure sample was obtained after trituration with hot methanol.

MS (APCI) 291 (M + H, 100%).

(b) [[6-Amino-1,4-dihydro-4-oxo-2 - [(phenylmethyl) thio] -5-pyrimidinyl] thio] -acetic acid ethyl ester

To a suspension of the product from step a) (1.5 g) in dry ethanol (100 ml) was added sodium borohydride (0.570 g), and the resulting solution was allowed to stir for 15 min. Ethyl bromoacetate (0.570 ml) was added to this solution. The mixture was neutralized with concentrated hydrochloric acid, then evaporated to dryness and purified (SiO2, ethyl acetate: dichloromethane 1: 1 as eluent) to give the subtitle compound as a colorless solid (1.1 g).

MS (APCI) 352 (M + H, 100%).

(c) 2 - [(Phenylmethyl) thio] -1H-pyrimido [5,4-b] [1,4] thiazin-4.7 (6H, 8H) -dione

P-Toluenesulfonic acid (50 mg) was added to a solution of the product from step b) (0.30 g) in dry toluene (60 ml), and the solution was heated under reflux for 11 hours. The resulting solid was collected by filtration, washed with ether, and dried to give the subtitle compound as a colorless solid (0.290 g).

MS (APCI) 306 (M + H, 100%).

(d) 4-Chloro-2 - [(phenylmethyl) thio] -6H-pyrimido [5,4-b] [1,4] -thiazin-7 (8H) -one

A suspension of the product from step c) (1.5 g), phosphorus oxychloride (10 ml) and N, N-dimethyl-aniline (1 ml) was heated under reflux for 2 hours. The mixture was allowed to cool to room temperature, and was carefully poured into a saturated sodium bicarbonate solution, and stirred for 15 min. The crude product was extracted into ethyl acetate and purified (SiO2, dichloromethane as eluent) to give the subtitle compound (0.25 g).

MS (APCI) 324 (M + H +, 100%).

(e) 4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -2 - [(phemlmethyl) -thio] -6H-pyrimido [5,4-b] [1,4] thiazin-7 (8H) -one

The product from step e) (0.250 g) in NMP (5 ml) was treated with (R) -2-amino-1-propanol (0.116 g), and the reaction mixture was heated to 110 ° C for 2 hours . The mixture was evaporated to dryness, and the residue was purified (HPLC, C18 Symmetry column^®, 0.1% aqueous ammonium acetate: acetonitrile, 75:25 isocratic elution) to give the title compound (0.13 g).

MS: APCI 363 (M + H) ¹H NMR: δ (DMSO) 10.84 (1H, s), 7.47-7.19 (5H, m), 6.26 (1H, d), 4.78 (1H, t), 4.36 -4.19 (3H, m), 3.55-3.32 (4H, m), 1.12 (3H, d).

ES 2 206 402 T3

Example 2

2 - [[(2,3-Difluorofeml) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

a) 2 - [[(2,3-Difluorophenyl) methyl] thio] -4,6-pyrimidinediamine

4,6-Diamino-2-pyrimidintiol (7.3 g) was dissolved in DMSO (100 ml) at room temperature under a nitrogen atmosphere. Potassium tert-butoxide (1M in THF, 48.3 ml) was added followed by 2,3-difluorobenzyl bromide (10.0 g). The mixture was stirred for 2 hours at room temperature. The reaction mixture was then partitioned between ethyl acetate and ammonium chloride. The organic phase was washed with ammonium chloride (3x) and brine, then dried over magnesium sulfate and evaporated to give the subtitle product as a white solid (12.2 g).

MS: ADCI (+ ve) 269 (M + 1)

b) 2 - [[(2,3-Difluorophenyl) methyl] thio] -5-nitroso-4,6-pyrimidinediamine

The product from step (a) (2.5 g) was dissolved in acetic acid (150 ml), and the solution was cooled to 5 ° C. A solution of sodium nitrite (625 mg) in water (50 ml) was added dropwise resulting in a dark blue coloration. The reaction was stirred at room temperature for 30 minutes, during which time a pink solid precipitated from solution. This was isolated by filtration and washed with water, then dried at 50 ° C to give the subtitle product as a blue solid (4.14 g).

MS: ADCI (+ ve) 298 (M + 1) ¹H NMR: δ (DMSO) 4.44 (s, 2H), 7.13-7.54 (m, 3H), 8.13 (s, 1H), 8.51 (s, 1H), 9.10 (s, 1H), 10.18 (s, 1H).

c) 2 - [[(2,3-Difluorophenyl) methyl] thio] -4,5,6-pyrimidine-triamine

Na2S2O4 (5.4 g) was added portionwise to a suspension of the product from step (b) (2 g) in boiling water (40 ml). The suspension was allowed to cool and then 50% sulfuric acid was slowly added, and then the mixture was cooled to 0 ° C. The solid was isolated by filtration and washed with cold water, then dried over P₂OR₅ at 50 ° C to give the subtitle product as a yellow solid.

MS: ADCI (+ ve) 284 (M + 1) ¹H NMR: δ (DMSO) 4.33 (s, 2H), 6.42 (br s, 3H), 7.10-7.48 (m, 3H)

d) 4-Amino-2 - [[(2,3-difluorophenyl) methyl] thio] -7 (8H) -pteridinone

The product from step (c) (100 mg) was dissolved in a solution of sodium (0.05 g) in methanol (5 ml). This was allowed to stir for 15 min at room temperature, then ethyl glyoxalate (134 µ ^ was added to the mixture, which was allowed to stir for 12 h at room temperature.

Water (5 ml) was added, then concentrated hydrochloric acid was added slowly to acidify the solution to ~ pH 5 whereby a solid precipitated which was isolated by filtration and dried over P₂OR₅ at 50 ° C to produce a pale yellow solid (44.5 mg).

MS: ADCI (+ ve) 322 (M + 1) ¹H NMR: δ (DMSO) 4.18 (s, 2H), 7.11-7.58 (m, 3H), 7.84 (s, 1H), 12.69 (bs, 1H)

e) 4-Bromo-2 - [[(2,3-difluorophenyl) methyl] thio] -7 (8H) -pteridinone

The product from step (d) (6.0 g) was suspended in DMSO (90 ml), bromoform (60 ml) was added, and the mixture was heated to 100 ° C. Isopentyl nitrite (25 ml) was added, and the mixture was stirred for 5 min. The mixture was quenched in an ice bath, then evaporated to leave an oil. This was repeated three times. Acetonitrile (200 ml) was added, and the solid that separated was removed by filtration. The solvent was evaporated, and the residue was purified by flash chromatography, eluting with dichloromethane and then 5% ethyl acetate in dichloromethane, to give a yellow solid which was suspended in ether, and then collected. The solid was washed with ether and dried to give the subtitle compound as a colorless solid (8.74 g).

MS: APCI (-ve) 382/4 (MH), 382 (100%) ¹H NMR: δ (DMSO) 4.47 (s, 2H), 7.13-7.55 (m, 3H), 8.14 (s, 1H), 13.33 (bs, 1H)

f) 2 - [[(2,3-difluorophenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

The product from step (e) (8.7 g) was dissolved in N-methylpyrrolidinone (40 ml), and Hunigs base (7.9

ES 2 206 402 T3 ml) followed by D-alaninol (2.7 ml). The mixture was stirred at 100 ° C for 15 min. The cooled solution was poured into water, (11), and acidified with dilute hydrochloric acid. The solid which separated was collected, washed with water and air dried. Crystallization from acetonitrile gave the title compound as a pale yellow solid (7.4 g).

mp 215-217 ° C

MS: APCI (+ ve) 380 (M + H, 100%) ¹H NMR: δ (DMSO) 1.14 (d, 3H), 3.48 (m, 2H), 4.31 (m, 1H), 4.45 (dd, 2H) 4.82 (t, 1H) 7.15 (m, 1H), 7.33 (m, 1H), 7.47 (t, 1H), 7.76 (d, 1H), 7.83 (d, 1H), 12.70 (s , 1 HOUR).

Alternatively, Example 2 can be prepared by the following procedure:

g) 2,4-bis [[(2,3-Difluorophenyl) methyl] thio] -7 (8H) -pteridinone

Sodium (3.96 g) was dissolved in methanol (150 ml), 5,6-diamino-2,4-pyrimidinedithiol (15 g) was added, then 2,3-difluorobenzyl bromide (30.9 g) was slowly added ), and the reaction mixture was stirred at room temperature under nitrogen for 10 min. Ethyl glyoxalate (15 ml) was added, followed by more sodium (2.5 g), and the reaction was left for another 20 min. The reaction was then quenched with acetic acid (10 ml), and poured into water (600 ml) with stirring. The resulting precipitate was filtered through celite, and washed with water. The filtrate was discarded, and the solid was washed through celite using acetone. The solution was then evaporated to dryness, and purified by silica gel column chromatography using 10% ethyl acetate in DCM to give the subtitle compound as a cream solid (8 g).

MS: APCI (+ ve) 465 (M + 1)

h) 2 - [[(2,3-Difluorophenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

The product from Example 2, step (g) (2.3 g), and D-alaninol (5 g) were microwaved at 150 ° C for 5 min. The resulting solution was partitioned between ethyl acetate and aqueous ammonium chloride, and the organic layer was washed with ammonium chloride (2x50 ml). The organic layer was evaporated to dryness and purified twice by silica gel chromatography using first DCM: methanol 20: 1 and then DCM: ethyl acetate 1: 1 to give the title compound (220mg).

MS: APCI (+ ve) 380 (M + 1)

Example 3

2 - [[(2,3-Difluorophenyl) methyl] thio] -4 - [[2-hydroxy-1- (hydroxymethyl) ethyl] amino] -7 (8H) -pteridinone

a) 4-Amino-2 - [[(2,3-difluorophenyl) methyl] thio] -7 (8H) -pteridinone

To a solution of triethyl phosphonoacetate (15.0 g) in tetrahydrofuran (60 ml), cooled in an ice bath, butyllithium (2.5 M in hexanes, 25.6 ml) was added at a rate such that the internal temperature was kept below 30 ° C. To this mixture was then added a solution of the product of Example 2, step (b) (10.0 g), in N, N-dimethylformamide (60 ml). The reaction mixture was refluxed for 1 hour, then cooled to room temperature, and quenched with acetic acid (6 ml). The solid thus precipitated was isolated by filtration, washed with water, ethanol and diethyl ether, and dried over P₂OR₅ at 50 ° C to give the subtitle product as a pale green solid (9.3 g).

MS: ADCI (+ ve) 322 (M + 1) ¹H NMR: δ (DMSO) 4.18 (s, 2H), 7.11-7.58 (m, 3H), 7.84 (s, 1H), 12.69 (bs, 1H)

b) 4-bromo-2 - [[(2,3-difluorophenyl) methyl] thio] -7 (8H) -pteridinone

The product from step (a) (0.5 g) was suspended in DMSO (10 ml) and bromoform (10 ml), and the mixture was heated to 125 ° C. Isoamyl nitrite (2 ml) was added, and the mixture was stirred at 125 ° C for 5 minutes before cooling in an ice bath. The solvent was removed by evaporation under high vacuum, and the residue was suspended in dichloromethane (100 ml). This suspension was washed with saturated aqueous ammonium chloride (50 ml), and then filtered through a plug of celite. The filtrate was evaporated and purified by column chromatography, eluting with 10% ethyl acetate in dichloromethane to give the subtitle compound as a white solid (0.22 g).

MS: ADCI (+ ve) 386 (M + 1) ¹H NMR: δ (DMSO) 4.47 (s, 2H), 7.13-7.55 (m, 3H), 8.14 (s, 1H), 13.33 (bs, 1H)

ES 2 206 402 T3

c) 2 - [[(2,3-Difluorophenyl) methyl] thio] -4 - [[2-hydroxy-1- (hydroxymethyl) ethyl] amino] -7 (8H) -pteridinone

The title compound (57 mg) was prepared from the product of step (b) (80 mg) and 2-amino-1,3-propanediol (29 mg), using the method of Example 2, step (f) .

MS: ADCI (+ ve) 396 (M + 1) ¹H NMR: δ (DMSO) 12.73 (1H, br s), 7.85 (1H, s), 7.50 (2H, m), 7.33 (1H, m), 7.15 (1H, m), 4.80 (2H, t), 4.45 (2H, s), 4.23 (1H, m), 3.55 (4H, m).

Example 4

2 - [[(2,3-Difluorophenyl) methyl] thio] -4 - [(2-hydroxyethyl) -amino] -7 (8H) -pteridinone

The title compound (52 mg) was prepared from the product of Example 3 step (b) (150 mg) and ethanolamine (35 µ / l), using the method of Example 2, step (f).

MS: ADCI (+ ve) 366 (M + 1) ¹H NMR: δ (DMSO) 12.71 (1H, br s), 8.09 (1H, brt), 7.84 (1H, s), 7.47 (1H, m), 7.32 (1H, m), 7.16 (1H, m), 4.78 (1H, t), 4.45 (2H, s), 3.53 (4H, m).

Example 5 (2R) -2 - [[2 - [[2,3-Difluorophenyl) methyl] thio] -7,8-dihydro-7-oxo-4-pteridinyl] amino] -propanamide

The title compound (160 mg) was prepared from the product of Example 3 step (b) (500 mg) and D-alaninamide hydrochloride (172 mg), using the method of Example 2 step (f).

MS: ADCI (+ ve) 393 (M + 1) ¹H NMR: δ (DMSO) 12.77 (1H, br s); 7.88 (1H, s), 7.85 (1H, d), 7.55 (1H, br s), 7.47 (1H, t), 7.36 (1H, m), 7.19 ( 1H, br s), 7.16 (1H, m), 4.60 (1H, q), 4.46 (2H, q), 1.40 (3H, d).

Example 6

2 - [[(3-Chloro-2-fluorophenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

a) 4-Amino-2 - [(phenylmethyl) thio] -7 (8H) -pteridinone

The subtitle compound (16.5 g) was prepared by the method of Example 3, step (a), using 5-nitroso-2 [(phenylmethyl) thio] -4,6-pyrimidinediamine (22.5 g).

MS (ESI) 286 (M + H, 100%).

b) 4-Amino-2 - [(phenylmethyl) sulfonyl] -7 (8H) -pteridinone

A suspension of the product of Example 6, step (a) (5.00 g), in acetic acid (500 ml) and peracetic acid (36-40% by weight solution in acetic acid, 50 ml) was stirred at 50 ° for 1 hour. After quenching with dimethyl sulfide (15 ml), the solution was poured into ice water (5000 ml). The suspended solid was removed by filtration, and dried in vacuo to give the subtitle compound as a pale yellow powder (4.53 g).

MS (ESI) 316 (MH, 100%).

c) 3-Chloro-2-fluoro-benzenemethanethiol

3-chloro-2-fluorobenzyl bromide (1.00 g) and thiourea (0.35 g) were heated together under reflux in ethanol (10 ml), for 1 hour. The solvent was removed in vacuo, and the residue was suspended in 10% aqueous sodium hydroxide (10 ml), and heated under reflux for 3 hours. The solution was then acidified with conc. HCl, and extracted into ether. Concentration of the organic phase in vacuo gave the subtitle compound as a pale yellow oil (0.71 g).

MS (EI) 176/178 (M +), 143/145 (100%).

d) 4-Amino-2 - [[(3-chloro-2-fluorophenyl) methyl] thio] -7 (8H) -pteridinone

The product of Example 6, step (c) (0.63 g), and a 1M solution of t-butoxide in THF (3.5 ml) were added to a suspension of the product of Example 6, step (b) (1 , 11 g) in NMP (20 ml). The mixture was vigorously stirred

ES 2 206 402 T3 for 1 hour, poured into 1M HCl (15 ml), and diluted with water. The suspended solid was filtered off and dried in vacuo to give the subtitle compound as a pale yellow powder (1.14 g).

MS (ESI) 338 (M + H, 100%).

e) 4-Bromo-2 - [[(3-chloro-2-fluorophenyl) methyl] thio] -7 (8H) -pteridinone

The subtitle compound (0.318 g) was prepared by the method of Example 3, step (b) using the product of Example 6, step (d) (1.10 g).

MS (ESI) 399/401 (MH, 100%).

f) 2 - [[(3-Chloro-2-fluorophenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

The title compound was prepared by the method of Example 2, step (f), using the product of Example 6, step (e) (75 mg). The reaction mixture was poured into water, and acidified with conc. HCl. The suspended solid was filtered off, air dried and recrystallized from acetonitrile to give a yellow solid (37 mg).

mp 209-211 °

MS (APCI) 396 (M + H, 100%).

¹H NMR: δ (DMSO) 12.71 (1H, br s), 7.83 (1H, s), 7.76 (1H, d), 7.64 (1H, t), 7.48 (1H, t), 7.17 (1H, t), 4.82 (1H, t), 4.39-4.48 (2H, m), 4.28 (1H, m), 3.40-3.52 (2H, m), 1.13 (3H, d).

Example 7

2 - [[(3-Chloro-2-fluorophenyl) methyl] thio] -4 - [(2-hydroxyethyl) -amino] -7 (8H) -pteridinone

The title compound (51 mg) was prepared by the method of Example 6, step (f), using the product of Example 6, step (e) (75 mg), and ethanolamine (25 µ / l).

Mp 195 - 196.5 °

MS (APCI) 382 (M + H, 100%).

¹H NMR: δ (DMSO) 12.71 (1H, br s), 8.08 (1H, br m), 7.84 (1H, s), 7.64 (1H, t), 7.49 (1H , t), 7.17 (1H, t), 4.79 (1H, brm), 4.44 (2H, s), 3.53 (4H, brm).

Example 8

2 - [[(3-Chloro-2-fluorophenyl) methyl] thio] -4 - [[2-hydroxy-1- (hydroxymethyl) ethyl] amino] -7 (8H) -pteridinone

The title compound (51 mg) was prepared by the method of Example 6, step (f), using the product of Example 6, step (e) (75 mg), and 2-amino-1,3-propanediol (37 mg).

mp 218.5 - 220.5 °

MS (APCI) 412 (M + H, 100%).

¹H NMR: δ (DMSO) 12.74 (1H, br s), 7.85 (1H, s), 7.67 (1H, t), 7.50 (2H, m), 7.16 (1H, t), 4.81 (2H, t), 4.44 (2H, s), 4.24 (1H, m), 3.51-3.62 (4H, m).

Example 9

[(2R) -2 - [[2 - [[(3-chloro-2-fluorophenyl) methyl] thiol-7,8-dihydro-7-oxo-4-pteridinyl] -amino 1,1-dimethylethyl ester ] propyl] carbamic (a) [(1R) -2-amino-1-methyl-2-oxoethyl] carbamic acid 9H-fluoren-9-ylmethyl ester

A solution of D-alaninamide hydrochloride (3 g), in 10% solution of sodium carbonate (50 ml) and dioxane (50 ml), was treated with FMOC chloride (6.24 g) in dioxane (40 ml ), and stirred overnight. The mixture was diluted with water (500 ml), and the product was collected by filtration and dried in vacuo to give 9.0 g of the subtitle compound.

MS (ESI) BP 311 (+ H)

ES 2 206 402 T3 (b) [(1R) -2-amino-1-methylethyl] carbamic acid 9H-fluoren-9-ylmethyl ester

Borane / dimethyl sulfide complex (4.4 ml) was added to a solution of the product of Example 9, step (a) (6.9 g) in THF (100 ml), and the mixture was heated under reflux for 2 hours. The mixture was carefully quenched by the addition of methanol (100 ml), evaporated to dryness, and the residue was taken up in methanol (100 ml) and acidified to pH 1-2 with concentrated hydrochloric acid. It was refluxed for 30 min, and then evaporated to dryness. The residue was triturated with ether to give a solid, which was collected by filtration, dissolved in water, and the free base was precipitated by the addition of aqueous sodium bicarbonate solution to give the subtitle compound (3.1 g).

MS (ESI) BP297 (+ H) (c) (2R) - [2- (9H-fluoren-9-ylmethoxycarbonylamne) -pn) pil] carbdmco acid 1,1-dinethylethyl ester

Ditert-butyl dicarbonate (2.2 g) was added to a stirred solution of the product of Example 9, step (b) (3.0 g), in THF (100 ml), and the mixture was stirred at room temperature for 30 min. The mixture was evaporated to dryness, and the crude product was purified (SiO2 dichloromethane as eluent) to give the subtitle compound (3.8 g).

NMR δH (CDCl₃) 7.76 (2H, m), 7.42 (2H, m), 7.39-26 (4H, s), 5.01 (1H, s), 4.85 (1H, s), 4, 38 (2H, d), 4.19 (1H, t), 3.77 (1H, m), 3.18 (2H, m), 1.27 (9H, s).

(d) [(2R) -2-αnino-propyl] carbanic acid 1,1-dimethylethic ester

Piperidine (5 ml) was added to a solution of the product from Example 9, step (c) (3.8 g), in THF (100 ml), and the mixture was allowed to stand for 1 hour at room temperature. The mixture was evaporated to dryness, and the residue was purified (SiO2, 5% methanol: dichloromethane as eluent) to give the subtitle compound as a colorless oil (1.7g).

NMR δH (CDCl₃) 4.95 (1H, s), 3.13 (1H, m), 2.99 (1H, m), 2.87 (1H, m), 1.38 (9H, s), 1.08 ( 3H, d).

(e) [(2R) -2 - [[2 - [[(3-chloro-2-fluorophenyl) methyl] thio] -7,8-dihydro-7-oxo-4- pteridinyl] amino] propyl] -carbamic

The title compound (83 mg) was prepared by the method of Example 6, step (f), using the product of Example 6, step (e) (75 mg) and the product of Example 9, step (d) (72 mg).

MS (APCI) 495 (M + H, 100%).

Ή NMR: δ (DMSO) 7.95 (1H, br d), 7.83 (1H, s), 7.62 (1H, t), 7.48 (1H, t), 7.17 (1H, t), 6.94 (1H, br t), 4.34-4.53 (3H, m), 3.04-3.17 (2H, m), 1.32 (9H, s), 1, 11 (3H, d).

Example 10

4 - [[(1R) -2-amno-1-netylethyl] amno] -2 - [[(3-chloro-2-fluorophenyl) nethyl] thio] -7 (8H) -pteridinone monohydrochlorum

A two phase mixture of the product from Example 9 (76 mg) in 1,4-dioxane (3 ml) and conc. HCl was stirred for 2 hours. (0.3 ml), then diluted with water (10 ml), and lyophilized. The residue was dissolved in water (10 ml), washed with ethyl acetate (5 ml), and lyophilized to give the title compound as a pale yellow solid (58 mg).

MS (APCI) 395 (M + H parent amine, 100%).

¹H NMR: δ (D2O) 7.86 (1H, s), 7.35 (1H, br t), 7.18 (1H, br t), 6.95 (1H, br t), 4.62 ( 1H, m), 4.20-4.41 (2H, m), 3.14-3.29 (2H, m), 1.33 (3H, brd).

Example 11

2 - [[(3-ChlorΌ-4-netoxyphenyl) nethyl] thio] -4 - [[2-hydroxy-1- (hydroxynethyl) ethyl] anino] -7 (8H) -pteridinone

a) 4-Amno-2 - [[(3-chlorΌ-4-nexyphenyl) nethyl] thio] -7 (8H) -pteridinone

The subtitle compound (1.36 g) was prepared by the method of Example 6, step (d), using the product of Example 6, step (b) (1.21 g), and 3-chloro-4-methoxybenzene -methanethiol (0.72 g).

MS (ESI) 350 (M + H, 100%).

ES 2 206 402 T3

b) 4-Bromo-2 - [[(3-chloro-4-methoxyphenyl) methyl] thio] -7 (8H) -pteridinone

The subtitle compound (0.28 g) was prepared by the method of Example 3, step (b), using the product of Example 11, step (a) (1.25 g).

MS (ESI) 411/413 (MH, 100%).

c) 2 - [[(3-Chloro-4-methoxyphenyl) methyl] thio] -4 - [[2-hydroxy-1- (hydroxymethyl) ethyl] amino] -7 (8H) -pteridinone

The title compound (89 mg) was prepared by the method of Example 8, using the product of Example 11, step (b) (0.100 g).

mp 209-211 °

MS (APCI) 424 (M + H, 100%).

¹H NMR: δ (DMSO) 12.73 (1H, s), 7.85 (1H, s), 7.54 (1H, d), 7.45 (2H, m), 7.06 (1H, d ), 4.81 (2H, br), 4.33 (2H, s), 4.20-4.28 (1H, m), 3.82 (3H, s), 3.51-3.63 ( 4H, m).

Example 12

4 - [(2-aminoethyl) amino] -2 - [[(3-chloro-4-methoxyphenyl) methyl] thio] -7 (8H) -pteridinone monotrifluoroacetate

The title compound (15 mg) was prepared by the method of Example 2, step (f), using the product of Example step (b) (50 mg), and ethylenediamine (24 µΐ). The reaction mixture was diluted with water (150 ml), lyophilized to a solid, and purified (HPLC, C8 Symmetry column^®, 0.1% aqueous trifluoroacetic acid: acetonitrile, isocratic elution 75:25).

MS (APCI) 393 (M + H parent amine, 100%).

¹H NMR: δ (D2O) 7.89 (1H, s), 7.39 (1H, s), 7.28 (1H, d), 6.92 (1H, d), 4.23 (2H, s ), 3.84 (5H, m), 3.25 (2H, br t). Example 13

2 - [[(2-Fluoro-4-methoxypheml) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinma

a) 2 - [[(2,3-Difluorophenyl) methyl] sulfonyl] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

2 - [[(2,3-Difluorophenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone (0.38 g) was stirred in acetonitrile (150 ml) and water (150 ml) with oxone (3.79 g) for 18 h. Acetonitrile was removed in vacuo to leave an aqueous suspension. The solid was collected and dried in vacuo to give the subtitle compound (0.30 g).

MS (APCI) 412 (M + H) (100%)

b) 2-Fluoro-4-methoxy-benzenemethanethiol

The subtitle compound (0.33 g) was prepared by the method of Example 6, step (c), using 1- (chloromethyl) -2-fluoro-4-methoxy-benzene (0.56 g).

MS (EI) 172 (M +), 139 (100%).

c) 2 - [[(2-Fluoro-4-methoxypheml) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] ammo] -7 (8H) -pteridinone.a

The title compound was prepared by the method of Example 6, step (d), using the product of Example 13, step (a) (0.20 g), the product of Example 13, step (b) (0.10 g), and DMSO (2 ml) instead of NMP as solvent. The reaction mixture was purified (HPLC, C8 Symmetry column^®, 0.1% aqueous ammonium acetate: acetonitrile, elution gradient 80:20 to 20:80) to give a white solid (63 mg).

MS (APCI) 392 (M + H, 100%).

¹H NMR: δ (DMSO) 12.69 (1H, br s), 7.83 (1H, s); 7.75 (1H, br d), 7.53 (1H, t), 6.83 (1H, d), 6.72 (1H, d), 4.83 (1H, br t), 4.34 (3H, m), 3.75 (3H, s), 3.41-3.54 (2H, m), 1.16 (3H, d).

ES 2 206 402 T3

Example 14

2- [[(2-Fluoro-3-methylphenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

a) 2-Fluoro-3-methyl-benzenemethanethiol

The subtitle compound (0.36 g) was prepared by the method of Example 6, step (c), using 2-fluoro-3-methylbenzyl bromide (0.55 g).

MS (El) 156 (M +), 123 (100%).

b) 2 - [[(2-Fluoro-3-methylphenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

The title compound (56 mg) was prepared by the method of Example 13, step (c), using the product of Example 13, step (a) (0.20 g), and the product of Example 14, step (a) (94 mg).

MS (APCI) 376 (M + H, 100%).

¹H NMR: δ (DMSO) 12.69 (1H, br s), 7.83 (1H, s), 7.76 (1H, d), 7.43 (1H, t), 7.18 (1H, t), 7.02 (1H, t), 4.83 (1H, t), 4.28-4.43 (3H, m), 3.41-3.54 (2H, m), 2.23 (3H, s), 1.15 (3H, d).

Example 15

2 - [[(3-Fluoro-2-methoxyphenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

a) 1- (Chloromethyl) -3-fluoro-2-methoxy-benzene

A solution of 3-fluoro-2-methoxy-benzenemethanol (WO 20000419) (0.50 g) and thionyl chloride (0.47 ml) in dichloromethane (30 ml) was stirred for 2 hours, and then concentrated to vacuum to give the subtitle compound, which was used directly in the next step.

b) 3-Fluoro-2-methoxy-benzenemethanethiol

The subtitle compound (0.31 g) was prepared by the method of Example 6, step (c), using the product of Example 15, step (a).

MS (EI) 172 (M +), 139 (100%).

c) 2 - [[(3-Fluoro-2-methoxyphenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

The title compound (84 mg) was prepared by the method of Example 13, step (c), using the product of Example 13, step (a) (0.20 g), and the product of Example 15, step (b) (0.10 g).

MS (APCI) 392 (M + H, 100%).

¹H NMR: δ (DMSO) 12.69 (1H, br s), 7.83 (1H, s), 7.76 (1H, d), 7.37 (1H, d), 7.18 (1H, m), 7.03 (1H, m), 4.83 (1H, br t), 4.31-4.42 (3H, m), 3.91 (3H, s), 3.42-3, 53 (2H, m), 1.16 (3H, d).

Example 16

2 - [[[4- (Difluoromethoxy) phenyl] methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

a) 4- (Difluoromethoxy) -benzenemethanethiol

The subtitle compound (0.20 g) was prepared by the method of Example 6, step (c), using 4 (difluoromethoxy) benzyl bromide (0.53 g).

MS (EI) 190 (M +), 107 (100%).

b) 2 - [[[4- (Difluoromethoxy) phenyl] methyl]] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

The title compound (51 mg) was prepared by the method of Example 13, step (c), using the product of Example 13, step (a) (0.20 g), and the product of Example 16, step (a) (0.17 g).

MS (APCI) 410 (M + H, 100%).

¹H NMR: δ (DMSO) 12.67 (1H, br s), 7.82 (1H, s), 7.74 (1H, d), 7.53 (2H, d), 7.19 (1H, t), 7.10 (2H, d), 4.82 (1H, t), 4.28-4.41 (3H, m), 3.39-3.52 (2H, m), 1.15 (3H, d).

ES 2 206 402 T3

Example 17

4 - [[(1R) -2-Hydroxy-1-methylethyl] amino] -2 - [[(4-hydroxyphenyl) -methyl] thio] -7 (8H) -pteridinone

The title compound (8 mg) was obtained as a by-product during the preparation of Example 16.

MS (APCI) 360 (M + H, 100%).

¹H NMR: δ (DMSO) 12.60 (1H, br), 9.35 (1H, s), 7.81 (1H, s), 7.70 (1H, d), 7.24 (2H, d ), 6.68 (2H, d), 4.83 (1H, t), 4.23-4.36 (3H, m), 3.44-3.55 (2H, m), 1.17 ( 3H, d).

Example 18

4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -2 - [[(3-methylphenyl) -methyl] thio] -7 (8H) -pteridinone

The title compound (75 mg) was prepared by the method of Example 13, step (c), using the product of Example 13, step (a) (0.20 g), and 3-methyl-benzeneomethane-thiol (75 mg).

MS (APCI) 358 (M + H, 100%).

¹H NMR: δ (DMSO) 12.66 (1H, br s), 7.82 (1H, s), 7.73 (1H, br d), 7.16-7.26 (3H, m), 7 .05 (1H, br d), 4.82 (1H, br t), 4.34 (3H, m), 3.40-3.54 (2H, m), 2.28 (3H, s), 1.17 (3H, br d).

Example 19

2 - [(1,3-Benzodioxol-4-ylmethyl) thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

a) 1,3-Benzodioxole-4-methanethiol

The subtitle compound (0.29 g) was prepared by the method of Example 6, step (c), using 4- (bromomethyl) -1,3-benzodioxole (0.51 g).

MS (EI) 168 (M +), 135 (100%).

b) 2 - [(1,3-Benzodioxol-4-ylmethyl) thio] 4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

The title compound (57 mg) was prepared by the method of Example 13, step (c), using the product of Example 13, step (a) (0.20 g), and the product of Example 19, step ( a) (0.10 g).

MS (APCI) 388 (M + H, 100%).

¹H NMR: δ (DMSO) 12.66 (1H, br s), 7.82 (1H, s), 7.75 (1H, br d), 6.99 (1H, d), 6.80 (2H , m), 6.04 (2H, s), 4.82 (1H, t), 4.33 (3H, m), 3.41-3.55 (2H, m), 1.16 (3H, d).

Example 20

2 - [[(2,4-Difluorofeml) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] ammo] -7 (8H) -pteridmone

a) 2,4-Difluoro-benzenemethanethiol

The subtitle compound (0.35 g) was prepared by the method of Example 6, step (c), using 2,4-difluoro-benzyl bromide (0.55 g).

MS (EI) 160 (M +), 127 (100%).

b) 2 - [[(2,4-Difluorofeml) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amine] -7 (8H) -pteridinone

The title compound (77 mg) was prepared by the method of Example 13, step (c), using the product of Example 13, step (a) (0.20 g), and the product of Example 20, step (a ) (87 mg).

MS (APCI) 380 (M + H, 100%).

¹H NMR: δ (DMSO) 12.70 (1H, br), 7.83 (1H, s), 7.67-7.76 (2H, m), 7.24 (1H, br t), 7, 03 (1H, br t), 4.82 (1H, br),

4.30-4.37 (3H, m), 3.43-3.49 (2H, m), 1.15 (3H, br d).

ES 2 206 402 T3

Example 21

2 - [[(3-Chlorophenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

The title compound (66 mg) was prepared by the method of Example 13, step (c), using the product of Example 13, step (a) (0.20 g), and 3-chloro-benzeneomethane-thiol (92 mg) .

MS (APCI) 378 (M + H, 100%).

¹H NMR: δ (DMSO) 12.70 (1H, br s), 7.82 (1H, s), 7.74 (1H, br d), 7.55 (1H, s), 7.45 (1H , br d), 7.34 (2H, m), 4.81 (1H, br t), 4.31-4.43 (3H, m), 3.43-3.50 (2H, m), 1.15 (3H, br d).

Example 22

4 - [[(1R) -2-Hydroxy-1-methylethyl] amino] -2 - [(5-isoxazolyl-methyl) thio] -7 (8H) -pteridinone

The title compound (0.038 g) was prepared by the method of Example 13, step (c), using the product of Example 13, step (a) (0.10 g), and 5-isoxazolylmethyl-mercaptan (0.057 g) .

Mp 191-194 ° C

MS (APCI) 335 (M + H, 100%).

¹H NMR: δ (DMSO) 12.71 (1H, br s); 8.47 (1H, s), 7.84 (1H, s), 7.80 (1H, d), 6.51 (1H, s), 4.81 (1H, t), 4.54 (2H , q), 4.26 (1H, m), 3.44 (2H, m), 1.13 (3H, d).

Example 23

4 - [[- 2-Hydroxy-1- (hydroxymethylethyl] amino] -2 - [(5-isoxazolyl-methyl) thio] -7 (8H) -pteridinone

a) 2 - [[(2,3-Difluorophenyl) methyl] sulfonyl] -4 - [[- 2-hydroxy-1-hydroxymethylethyl] amino] -7 (8H) -pteridinone

2 - [[(2,3-Difluorophenyl) methyl] thio] -4 - [[- 2-hydroxy-1-hydroxymethylethyl] amino] -7 (8H) -pteridinone (2.25 g) was stirred in acetonitrile (750 ml) and water (750 ml) with oxone (22.5 g) for 18 h. Acetonitrile was removed in vacuo to leave an aqueous suspension. The mixture was extracted into ethyl acetate. The combined extracts were washed with saturated sodium chloride solution, dried (MgSO4), and the solvent was removed in vacuo to give the subtitle compound (1.92g).

MS (APCI) 428 (M + H) (100%)

b) 4 - [[- 2-Hydroxy-1- (hydroxymethylethyl] amino] -2 - [(5-isoxazolylmethyl) thio] -7 (8H) -pteridinone

The title compound (0.03 g) was prepared by the method of Example 13, step (c), using the product of Example 23, step (a) (0.20 g), and 5-isoxazolylmethyl-mercaptan (0, 1 g).

Mp 199-203 ° C

MS (APCI) 351 (M + H, 100%).

¹H NMR: δ (DMSO) 8.47 (1H, s), 7.86 (1H, s), 7.54 (1H, d), 6.55 (1H, s), 4.80 (2H, t ), 4.56 (2H, s), 4.22 (1H, m), 3.55 (4H, m).

Example 24

4 - [[(1R) -2-Hydroxy-1-methylethyl] amino] -2 - [[(5-methyl-2-furanyl) methyl] thio] -7 (8H) -pteridinone

The title compound (0.058 g) was prepared by the method of Example 13, step (c), using the product of Example 13, step (a) (0.20 g), and 5-methyl-2-furanylmethyl-mercaptan (0.15 g of 80% purity).

Mp 197-199 ° C

MS (APCI) 348 (M + H, 100%).

¹H NMR: δ (DMSO) 12.65 (1H, br s); 7.82 (1H, s), 7.74 (1H, d), 6.27 (1H, d), 5.96 (1H, d), 4.83 (1H, t), 4.37 (2H ,

s), 4.32 (1H, m), 3.47 (2H, m), 2.22 (3H, s), 1.17 (3H, d).

ES 2 206 402 T3

Example 25

2 - [[(2-Fluorophenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

The title compound (0.089 g) was prepared by the method of Example 13, step (c), using the product of Example 13, step (a) (0.20 g), and 2-fluorobenzyl mercaptan (0.083 g) .

Mp 203-205 ° C

MS (APCI) 362 (M + H, 100%).

¹H NMR: δ (DMSO) 12.69 (1H, br s), 7.83 (1H, s), 7.75 (1H, d), 7.64 (1H, m), 7.32 (1H , m), 7.20 (1H, m), 7.14 (1H, m), 4.82 (1H, t), 4.41 (2H, q), 4.31 (1H, m), 3 , 47 (2H, m), 1.15 (3H, d).

Example 26

4 - [[(1R) -2-Hydroxy-1-methylethyl] amino] -2 - [[(2-thienyl) -methyl) thio] -7 (8H) -pteridinone

The title compound (0.076 g) was prepared by the method of Example 13, step (c), using the product of Example 13, step (a) (0.20 g), and 2-thienylmethyl mercaptan (0.050 g).

Mp 209-212 ° C

MS (APCI) 350 (M + H, 100%).

¹H NMR: δ (DMSO) 12.66 (1H, br s), 7.83 (1H, s), 7.77 (1H, d), 7.37 (1H, d), 7.12 (1H, d), 6.93 (1H, dd), 4.82 (1H, t), 4.62 (2H, q), 4.36 (1H, m), 3.46 (2H, m), 1, 17 (3H, d).

Example 27

2 - [[(2-Fluorophenyl) methyl] thio] -4 - [[- 2-hydroxy-1- (hydroxy-methylethyl] amino] -7 (8H) -pteridinone

The title compound (0.088 g) was prepared by the method of Example 13, step (c), using the product of Example 23 step (a) (0.21 g), and 2-fluorobenzylmercaptan (0.83 g).

Mp 206-208 ° C

MS (APCI) 378 (M + H, 100%).

¹H NMR: δ (DMSO) 12.67 (1H, br s), 7.84 (1H, s), 7.66 (1H, t), 7.47 (1H, d), 7.32 (1H, m), 7.16 (2H, m), 4.80 (2H, t), 4.41 (2H, s), 4.25 (1H, m), 3.57 (4H, m).

Example 28

4 - [[- 2-Hydroxy-1- (hydroxymethyl) ethyl] amino] -2 - [(2-thienylmethyl) thio] -7 (8H) -pteridinone

The title compound (0.075 g) was prepared by the method of Example 13, step (c), using the product of Example 23, step (a) (0.21 g), and 2-thienylmethyl mercaptan (0.05 g) .

Mp 220-223 ° C

MS (APCI) 366 (M + H, 100%).

¹H NMR: δ (DMSO) 12.67 (1H, br s), 7.85 (1H, s), 7.49 (1H, d), 7.38 (1H, d), 7.15 (1H, d), 6.93 (1H, dd), 4.81 (2H, t), 4.64 (2H, s), 4.28 (1H, m), 3.59 (4H, m).

Example 29

2 - [[(2-Fluoro-5-methylphenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

a) 2-Fluoro-5-methyl-benzenemethanethiol acetate

Diisopropyl azodicarboxylate (2.68 ml) was added to a solution of triphenylphosphine (3.57 g) in dry THF (30 ml) at 0 ° C. The mixture was stirred for 1 hr. A solution of 2-fluoro-5-methyl-benzenemethanol (0.96 ml) in dry THF (10 ml) was added dropwise over 40 min. The mixture was stirred for 1 hr, then warmed to room temperature, and stirred another 3 hr. The solvent was evaporated, and the residue was suspended with ether and filtered. The filtrate was evaporated and purified by column chromatography, eluting with 3% ethyl acetate in isohexane to give the subtitle compound as an oil (1.23 g).

ES 2 206 402 T3

GC / MS 100% (EI) 198 (M +), 123 (100%)

b) 2-Fluoro-5-methyl-benzylmethanethiol

2-Fluoro-5-methyl-benzeneomethane-thiol acetate (1.2 g) in methanol, previously saturated with ammonia (10 ml), was stirred at room temperature for 30 min. The solvent was evaporated, and the residue was partitioned between ether and water. The layers were separated, and the water was extracted with ether. The combined ether was washed with water, dried and evaporated to leave an oil (0.9 g).

GC / MS 97% (EI) 156 (M +) 123 (100%)

c) 2 - [[(2-Fluoro-5-methylphenyl) methyl] thio] 4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

The title compound (0.093 g) was prepared by the method of Example 13, step (c), using the product of Example 13, step (a) (0.20 g), and the product of Example 29, step (b ).

Mp 202-204 ° C

MS (APCI) 376 (M + H, 100%).

¹H NMR: δ (DMSO) 12.68 (1H, br s), 7.82 (1H, s), 7.74 (1H, d), 7.41 (1H, d), 7.07 (2H, m), 4.83 (1H, t), 4.33 (3H, m), 3.49 (2H, m), 2.25 (3H, s), 1.17 (3H, d).

Example 30

2 - [[(2-Fluoro-3- (trifluoromethyl) phenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

a) 4- (1-Methylethoxy) -2 - [[[3- (trifluoromethyl) phenyl] -methyl] thio] -7 (8H) -pteridinone

6 - [[[2-fluoro-3- (trifluoromethyl) phenyl] -methyl] thio] -2 - [[[3- (trifluoromethyl) phenyl] methyl] thio] -4,5-pyrimidinediamine (7.12 g) to a sodium methoxide solution prepared from sodium (0.62 g) and methanol (150 ml). Ethyl glyoxalate (6.5 ml of 50% in toluene) was added, and the mixture was refluxed for 30 min. The solvent was evaporated, and redissolved in 2-propanol. Sodium (1.1 g) was added, and the mixture was refluxed for 30 min. The solvent was evaporated, and the residue was taken up in water, and acidified with dilute hydrochloric acid. The mixture was extracted with dichloromethane, and the extracts were washed with water, then dried (MgSO4), and evaporated. Purification by flash chromatography eluting with 10% ethyl acetate in dichloromethane gave the subtitle compound as a solid (2.64 g).

Mp 205-206 ° C

MS (APCI) 415 (M + H, 100%) ¹H NMR: δ (DMSO) 13.03 (1H, s), 8.06 (1H, t), 7.97 (1H, s), 7.70 (1H, t), 7.38 (1H, t ), 5.38 (1H, m), 4.53 (2H, s), 1.31 (6H, d).

b) 2 - [[(2-Fluoro-3- (trifluoromethyl) phenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone

The product from Example 30, step (a) (0.3 g), and D-alaninol (0.6 ml) were sonicated with N-methylimidazole (1 ml) to form a paste. The pasta was heated in a 300W microwave at 160 ° C for 25 min. The solvent was removed by bulb-to-bulb distillation, and the residue was purified by reverse phase HPLC, to give the title compound (0.52 g).

Mp 217-219 ° C

MS (APCI) 430 (M + H, 100%) ¹H NMR: δ (DMSO) 8.03 (1H, t), 7.83 (1H, s), 7.76 (1H, d), 7.68 (1H, t), 7.35 (1H, t ) 4.82 (1H, t), 4.47 (2H, q), 4.26 (1H, m), 3.45 (2H, m), 1.12 (3H, d).

Example 31

2 - [[(2-Fluoro-3- (trifluoromethyl) phenyl) methyl] thio] -4 - [[2-hydroxy-1- (hydroxymethyl) ethyl] amino] -7 (8H) -pteridinone

The title compound (0.049 g) was prepared by the method of Example 30, step (b), using the product of Example 30, step (a) (0.30 g), and 2-aminopropane-1,3-diol (0.66 g).

Mp 244-245 ° C

ES 2 206 402 T3

MS (APCI) 446 (M + H, 100%).

¹H NMR: δ (DMSO) 12.76 (1H, s), 8.06 (1H, t), 7.85 (1H, s), 7.68 (1H, t), 7.52 (1H, d ), 7.35 (1H, t), 4.81 (2H, t), 4.47 (2H, s), 4.23 (1H, m), 3.55 (4H, m).

Example 32

2 - [[(2-Fluoro-3- (trifluoromethyl) phenyl) methyl] thio] -4 - [(2-hydroxyethyl) amino] -7 (8H) -pteridinone

The title compound (0.072 g) was prepared by the method of Example 30, step (b), using the product of Example 30, step (a) (0.30 g), and ethanolamine (0.5 ml)

Mp 221-222 ° C

MS (APCI) 416 (M + H, 100%).

¹H NMR: δ (DMSO) 12.73 (1H, s), 8.08 (1H, m), 7.03 (1H, t), 7.85 (1H, s), 7.68 (1H, t ), 7.36 (1H, t), 4.78 (1H, t), 4.47 (2H, s), 3.52 (4H, m).

Example 33

4 - [(2-Aminoethyl) amino] -2 - [[(2-fluoro-3- (trifluoromethyl) -phenyl) methyl] thio] -7 (8H) -pteridinone

The title compound (0.054 g) was prepared by the method of Example 30, step (b), using the product of Example 30, step (a) (0.30 g), and ethylenediamine (0.5 ml). The product was purified by methanol crystallization.

MS (APCI) 415 (M + H, 100%).

¹H NMR: δ (DMSO) 8.01 (1H, t), 7.91 (1H, br s), 7.67 (1H, m), 7.58 (1H, br s), 7.52 (1H , bs), 7.35 (1H, t), 7.17 (1H, br s), 4.45 (2H, s), 3.42 (2H, t), 3.15 (2H, br s) , 2.75 (8H, m).

Example 34

2 - [[(2,3-Difluorophenyl) methyl] thio] -4 - [[2-hydroxy-1- (hydroxymethyl) -1-methylethyl] amino] -7 (8H) -pteridinone

The product from Example 2, step (e) (400 mg), was dissolved in N-methylpyrrolidinone (15 ml), and Hunigs base (0.36 ml) was added followed by 2-amino-2-methyl-1, 3-propanediol (0.273 g). The mixture was stirred at 110 ° C for 17 hours. The resulting mixture was poured into water, and allowed to stand overnight to allow a solid to precipitate. This solid was collected by filtration, then purified by reverse phase preparative HPLC on a Symmetry C-8 column.^®, using 10 to 95% acetonitrile in 0.1% aqueous ammonium acetate, at 20 ml / min for 10 min, to give the title compound as off-white needles (0.113 g).

MS: APCI (+ ve) 410 (M + H) ¹H NMR: δ (DMSO) 1.30 (3H, s), 3.54-3.58 (2H, m), 3.63-3.67 (2H, m), 4.45 (2H, s) , 4.98-5.01 (2H, t), 7.13 (1H, s), 7.15-7.19 (1H, m), 7.31-7.36 (1H, m), 7 , 45-7.49 (1H, t), 7.84 (1H, s), 12.74 (1H, bs).

Example 35

2 - [[(2,3-Difluorophenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -6-methyl-7 (8H) -pteridinone

a) 4-Amino-2 - [[(2,3-difluorophenyl) methyl] thio] -6-methyl-7 (8H) -pteridinone

The subtitle compound was prepared from the product of Example 2, step (b) (2 g), and triethyl phosphono-propionate (3.2 g), using the method of Example 3, step (a).

MS: APCI (+ ve) 336 (M + 1) ¹H NMR: δ (DMSO) 2.33 (s, 3H), 4.40 (s, 2H), 7.09-7.58 (m, 3H), 12.56 (bs, 1H).

b) 4-Bromo-2 - [[(2,3-difluorophenyl) methyl] thio] -6-methyl-7 (8H) -pteridinone

The subtitled compound was prepared from the product of Example 35, step (a) (1.5 g), and bromoform (30 ml), using the method of Example 3, step (b).

MS: APCI (+ ve) 400 (M + 1) ¹H NMR: δ (DMSO) 2.39 (s, 3H), 4.52 (s, 2H), 7.11-7.55 (m, 3H), 13.21 (bs, 1H).

ES 2 206 402 T3

c) 2 - [[(2,3-Difluorophenyl) nethyl] thio] -4 - [[(1R) -2-hydroxy-1-netylethyl] anino] -6-nethyl-7 (8H) -pteridinone

The title compound was prepared from the product of Example 35, step (b) (200 mg), and D-alaninol (120 µ ^, using the method of Example 2, step (f).

MS: APCI (+ ve) 394 (M + 1) ¹H NMR: δ (DMSO) 1.12 (d, 3H), 2.34 (s, 3H), 3.45 (m, 2H), 4.24 4.33 (m, 1H), 4.43 ( s, 2H), 4.85 (bs, 1H), 7.107.49 (m, 3H), 12.58 (s, 1H).

Example 36

2 - [[(2,3-Difluorophenyl) nethyl] thio] -7,8-dihydro-4 - [[(1R) -2-hydroxy-1-netylethyl] anino] -7-oxo-6 acid ethyl ester -pteridinecarboxylic

a) 4-Anino-2 - [[(2,3-difluoro-phenyl) nethyl] thio] -7,8-dihydro-7-oxo-6-pteridinecarboxylic acid ethyl ester

The product from Example 2, step (b) (5 g), was dissolved in diethyl malonate (100 ml), and heated to 120 ° C for 10 h, with stirring. The reaction mixture was allowed to cool, and the solid that precipitated was filtered, washed with water, and oven dried at 50 ° C to give the product as a yellow solid (3.2 g).

MS: APCI (+ ve) 394 (M + 1) ¹H NMR: δ (DMSO) 1.29-1.33 (t, 3H), 4.28-4.35 (q, 2H), 4.42 (s, 2H) 7.11-7.59 (m , 3H), 7.88 (bs, 1H), 8.08 (bs, 1H), 13.05 (s, 1H).

b) 4-Brono-2 - [[(2,3-difluorophenyl) nethyl] thio] -7,8-dihydro-7-oxo-6-pteridine-carboxylic acid ethyl ester

The subtitled compound was prepared from the product of Example 36, step (a) (2 g), and bromoform (40 ml) using the method of Example 3, step (b).

MS: APCI (+ ve) 458 (M + 1) ¹H NMR: δ (DMSO) 1.31 (t, 3H), 4.40 (q, 2H), 4.52 (s, 2H), 7.13-7.55 (m, 3H), 13.05 (s, 1H).

c) 2 - [[(2,3-Difluorophenyl) -nethyl] thio] -7,8-dihydro-4 - [[(1R) -2-hydroxy-1-netylethyl] -anine] -7oxo acid ethyl ester -6-pteridinecarboxylic

The title compound was prepared from the product of Example 36, step (b) (300 mg), and D-alaninol (180 µ ^, using the method of Example 2, step (f).

MS: APCI (+ ve) 451 (M + 1) ¹H NMR: δ (DMSO) 1.12 (d, 3H), 1.29 (t, 3H), 3.41-3.53 (m, 2H), 4.22-4.32 (m, 3H) , 4.51 (s, 2H), 4.83-4.86 (t, 1H), 7.07-7.47 (m, 3H).

Example 37

2 - [[(2,3-Difluorophenyl) nethyl] thio] -4 - [[(1R) -2-hydroxy-1-netylethyl] anino] -6- (trifluoronethyl) -7 (8H) -pteridinone

a) 4-Anino-2 - [[(2,3-difluorophenyl) nethyl] thio] -6- (trifluoronethyl) -7 (8H) -pteridinone

The subtitle compound was prepared from the product of Example 2, step (c) (5 g), and trifluoropyruvate (10 ml), using the method of Example 2, step (d).

MS: APCI (+ ve) 390 (M + 1) ¹H NMR: δ (DMSO) 4.32 (s, 2H), 7.10-7.70 (m, 3H), 7.92-8.23 (2bs, 2H), 13.23 (bs, 1H) .

b) 4-Brono-2 - [[(2,3-difluorophenyl) nethyl] thio] -6- (trifluoronethyl) -7 (8H) -pteridinone

The subtitled compound was prepared from the product of Example 37, step (a) (1.5 g), and bromoform (30 ml), using the method of Example 3, step (b).

MS: APCI (+ ve) 454 (M + 1).

ES 2 206 402 T3

c) 2 - [[(2,3-Difluorophenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -6- (trifluoromethyl) -7 (8H) -pteridmone

The title compound was prepared from the product of Example 37, step (b) (150 mg), and D-alaninol (100 µ ^, using the method of Example 2, step (f).

MS: APCI (+ ve) 448 (M + 1) ¹H NMR: δ (DMSO) 1.15 (d, 3H), 3.50 (m, 2H), 4.37 (m, 1H), 4.47 (s, 2H), 4.89 (t, 1H ), 7.12-7.49 (m, 3H), 7.79 (d, 1H), 13.25 (bs, 1H).

Example 38

2 - [[(2,3-DifluorOphenyl) methyl] thio] -4 - [[(1R) -2-hydroxy-1-methylethyl] amino] -7 (8H) -pteridinone sodium salt

The product from Example 2, step (f), was suspended in water containing one equivalent of sodium hydroxide, followed by the addition of a small aliquot of tetrahydrofuran and methanol to aid dissolution. The resulting solution was then lyophilized to give the title compound as a colorless solid.

Mp 255-260 ° C (dec.)

MS: APCI (+ ve) 380 (M + 1) ¹H NMR: δ (DMSO) 7.42 (1H, m), 7.37 (1H, s), 7.31 (1H, m), 7.13 (1H, m), 6.89 (1H, d ), 4.79 (1H, t), 4.40 (2H, s), 4.15 (1H, m), 3.45 (2H, m), 1.12 (3H, d).

Examples 39-72

Examples 39a 72 were prepared by heating the product of Example 2, step (e) (2.5x10^-6 moles), with the appropriate amine (2 equivalents) and N-ethyldiisopropylamine (6 equivalents) in N-methylpyrrolidinone (0.25 ml) in a sealed vessel, at 100 ° C, for 1 hour.

Example 39

2 - [(2,3-difluon) benzyl) thio] -4- (ethylamino) -7 (8H) -pteridinone

MS: APCI (+ ve) 350 (M + 1).

Example 40

2 - [(2,3-difluorobenzyl) thio] -4- (isopropylamino) -7 (8H) -pteridinone

MS: APCI (+ ve) 364 (M + 1).

Example 41 (+/-) - 4- (sec-butylamino) -2 - [(2,3-difluorobenzyl) thio] -7 (8H) -pteridinone

MS: APCI (+ ve) 378 (M + 1).

Example 42

2 - ({2 - [(2,3-difluorobenzyl) thio] -7-oxo-7,8-dihydro-4-pteridinyl} amino) acetamide

MS: APCI (+ ve) 379 (M + 1).

Example 43 (+/-) - 2 - [(2,3-difluorobenzyl) thio] -4 - [(2-hydroxypropyl) -amino] -7 (8H) -pteridinone

MS: APCI (+ ve) 380 (M + 1).

Example 44 (S) -2 - [(2,3-Difluorobenzyl) thio] -4 - [(2-hydroxy-1-methyl-ethyl) amino] -7 (8H) -pteridinone

MS: APCI (+ ve) 380 (M + 1).

ES 2 206 402 T3

Example 45 (+/-) - 2 - [(2,3-difluorobenzyl) thio] -4 - [(2-hydroxy-1-methylethyl) amino] -7 (8H) -pteridinone

MS: APCI (+ ve) 380 (M + 1).

Example 46 (R) -2 - [(2,3-Difluorobenzyl) thio] -4 - [(2-hydroxypropyl) amino] -7 (8H) -pteridinone

MS: APCI (+ ve) 380 (M + 1).

Example 47

2 - [(2,3-difluorobenzyl) thio] -4 - [(3-hydroxypropyl) amino] -7 (8H) -pteridinone

MS: APCI (+ ve) 380 (M + 1).

Example 48

2- [(2,3-Difluorobenzyl) thio] -4 - [(2-hydroxyethyl) (methyl) -amino] -7 (8H) -pteridinone

MS: APCI (+ ve) 380 (M + 1).

Example 49

3- [{2 - [(2,3-difluorobenzyl) thio] -7-oxo-7,8-dihydro-4-pteridinyl} (methyl) amino] propane nitrile

MS: APCI (+ ve) 389 (M + 1).

Example 50 (R) -2 - [(2,3-Difluorobenzyl) thio] -4 - {[1- (hydroxymethyl) -propyl] amino} -7 (8H) -pteridinone

MS: APCI (+ ve) 394 (M + 1).

Example 51 (S) -2 - [(2,3-Difluorobenzyl) thio] -4 - {[1- (hydroxymethyl) -propyl] amino} -7 (8H) -pteridinone

MS: APCI (+ ve) 394 (M + 1).

Example 52

2 - [(2,3-difluorobenzyl) thio] -4 - [(4-hydroxybutyl) amino] -7 (8H) -pteridinone

MS: APCI (+ ve) 394 (M + 1).

Example 53 (+/-) - 2 - [(2,3-difluorobenzyl) thio] -4 - {[1- (hydroxymethyl) propyl] amino} -7 (8H) -pteridinone

MS: APCI (+ ve) 394 (M + 1).

Example 54

2 - [(2,3-difluorobenzyl) thio] -4 - [(2-hydroxy-1,1-dimethylethyl) amino] -7 (8H) -pteridinone

MS: APCI (+ ve) 394 (M + 1).

Example 55

2 - [(2,3-difluorobenzyl) thio] -4- [ethyl (2-hydroxyethyl) amino] -7 (8H) -pteridinone

MS: APCI (+ ve) 394 (M + 1).

ES 2 206 402 T3

Example 56 (+/-) - 4 - [(3-amino-2-hydroxypropyl) amino] -2 - [(2,3-difluoro-benzyl) thio] -7 (8H) -pteridinone

MS: APCI (+ ve) 395 (M + 1).

Example 57 (+/-) - 2 - [(2,3-difluorobenzyl) thio] -4 - [(1,3-dimethylbutyl) -amino] -7 (8H) -pteridinone

MS: APCI (+ ve) 406 (M + 1).

Example 58 (1R, 2R) -2 - [(2,3-difluorobenzyl) thio] -4 - [(2-hydroxy-cyclopentyl) amino] -7 (8H) -pteridinone

MS: APCI (+ ve) 406 (M + 1).

Example 59

2 - [(2,3-difluorobenzyl) thio] -4 - [(5-hydroxypentyl) amino] -7 (8H) -pteridinone

MS: APCI (+ ve) 408 (M + 1).

Example 60 (+/-) - 2 - [(2,3-difluorobenzyl) thio] -4 - {[1- (hydroxymethyl) butyl] amino} -7 (8H) -pteridinone

MS: APCI (+ ve) 408 (M + 1).

Example 61 (+/-) - 2 - ({2 - [(2,3-Difluorobenzyl) thio] -7-oxo-7,8-dihydro-4-pteridinyl} amino) methyl propanoate

MS: APCI (+ ve) 408 (M + 1).

Example 62

2 - [(2,3-Difluorobenzyl) thio] -4 - [(3-hydroxy-2,2-dimethylpropyl) amino] -7 (8H) -pteridinone

MS: APCI (+ ve) 408 (M + 1).

Example 63 (1R, 2R) -2 - [(2,3-difluorobenzyl) thio] -4 - {[2-hydroxy-1- (hydroxymethyl) propyl] amino} -7 (8H) -pteridinone

MS: APCI (+ ve) 410 (M + 1).

Example 64

4- [Bis (2-hydroxyethyl) aminol-2 - [(2,3-difluorobenzyl) thio] -7 (8H) -pteridinone

MS: APCI (+ ve) 410 (M + 1).

Example 65

2 - [(2,3-difluorobenzyl) thio] -4 - {[2- (2-hydroxyethoxy) ethyl] amino} -7 (8H) -pteridinone

MS: APCI (+ ve) 410 (M + 1).

Example 66

2 - [(2,3-Difluorobenzyl) thio] -4 - [(2,2-dimethoxyethyl) amino] -7 (8H) -pteridinone

MS: APCI (+ ve) 410 (M + 1).

ES 2 206 402 T3

Example 67

4 - {[2- (diethylamino) ethyl] amino} -2 - [(2,3-difluorobenzyl) thio] -7 (8H) -pteridinone

MS: APCI (+ ve) 421 (M + 1).

Example 68 (S) -2 - [(2,3-Difluorobenzyl) thio] -4 - {[1- (hydroxymethyl) -2,2-dimethylpropyl] amino} -7 (8H) -pteridinone

MS: APCI (+ ve) 422 (M + 1).

Example 69 (R) -2 - [(2,3-Difluorobenzyl) thio] -4 - {[1- (hydroxymethyl) -3-methylbutyl] amino} -7 (8H) -pteridinone

MS: APCI (+ ve) 422 (M + 1).

Example 70

2 - [(2,3-difluorobenzyl) thio] -4 - [(6-hydroxyhexyl) amino] -7 (8H) -pteridinone

MS: APCI (+ ve) 422 (M + 1).

Example 71

2 - [(2,3-Difluorobenzyl) thio] -4 - {[3- (dimethylamino) -2,2-dimethylpropyl] amino} -7 (8H) -pteridinone.

MS: APCI (+ ve) 436 (M + 1).

Example 72 (S) -2 - ({2 - [(2,3-difluorobenzyl) thio] -7-oxo-7,8-dihydro-4-pteridinyl} amino) -3-hydroxypropanoate

MS: APCI (+ ve) 438 (M + 1).

Ligand binding assay pharmacological data

The [¹²⁵I] IL-8 (human, recombinant) was purchased from Amersham, UK, with a specific activity of 2,000 Ci / mmol. All other chemicals were analytical grade. Elevated levels of hrCXCR2 were expressed in HEK 293 cells (ECACC human embryonic kidney 293 cells # 85120602) (Lee et al. (1992) J. Biol. Chem. 267, p. 16283-16291). The hrCXCR2 cDNA was amplified and cloned from human neutrophil mRNA. DNA was cloned into PCRScript (Stratagene), and clones were identified using DNA. The coding sequence was subcloned into the eukaryotic expression vector RcCMV (Invitrogen). Plasmid DNA was prepared using Quiagen Megaprep 2500, and transfected into HEK 293 cells using Lipofectamine reagent (Gibco BRL). The cells of the most highly expressed clone were harvested in phosphate buffered saline containing 0.2% (w / v) ethylenediaminetetraacetic acid (EDTA), and centrifuged (200 g, 5 min.). The cell pellet was resuspended in ice-cold homogenization buffer [10 mM HEPES (pH 7.4), 1 mM dithiothreitol, 1 mM EDTA, and a panel of protease inhibitors (1 mM phenylmethylsulfonyl fluoride, 2 μg / ml inhibitor of soybean trypsin, 3 mM benzamidine, 0.5 µg / ml of leupeptin and 100 µg / ml of bacitracin)], and the cells were allowed to swell for 10 minutes. The cell preparation was disrupted using a manual glass mortar type homogenizer with a PTFE pestle, and the cell membranes were harvested by centrifugation (45 minutes, 100,000 g, 4 ° C). The membrane preparation was stored at -70 ° C in homogenization buffer supplemented with Tyrode's salt solution (137 mM NaCl, 2.7 mM KCl, 0.4 mM NaH2PO4), 0.1% (w / v) of gelatin and 10% (v / v) glycerol.

All assays were performed in 96-well MultiScreen 0.45 µm filter plates (Millipore, UK). Each assay contained ~ 50 pM of [¹²⁵I] IL-8 and membranes (equivalent to ~ 200,000 cells) in assay buffer [Tyrode's salt solution supplemented with 10 mM HEPES (pH 7.4), 1.8 mM CaCl2, 1 mM MgCl2, 0.125 mg / ml of bacitracin and 0.1% (w / v) of gelatin]. Furthermore, a compound of formula (I) according to the Examples was predissolved in DMSO and added to reach a final concentration of 1% (v / v) of DMSO. The assay was started with the addition of the membranes and, after 1.5 hours at room temperature, the membranes were harvested by filtration using a Millipore MultiScreen vacuum filter, and washed twice with assay buffer (no bacitracin). The back plates were removed from the MultiScreen plate mount, the filters were dried at room temperature, perforated and then counted in a γ Cobra counter.

Compounds of formula (I) according to the Examples were found to have IC values₅₀ less than (&lt;) 10 μM.

ES 2 206 402 T3

Intracellular calcium mobilization assay

Human neutrophils were prepared from EDTA-treated peripheral blood, as previously described (Baly et al. (1997) Methods in Enzymology 287, p. 70-72), in storage buffer [Tyrode's salt solution (137 mM NaCl, 2.7 mM KCl, 0.4 mM NaH2PO4) supplemented with 5.7 mM glucose and 10 mM HEPES (pH 7.4)].

GROa (human, recombinant) chemokine was obtained from R&amp;D Systems (Abingdon, UK). All other chemicals were analytical grade. Changes in intracellular free calcium were measured fluorometrically by loading neutrophils with the calcium-sensitive fluorescent dye, fluo-3, as previously described (Merritt et al. (1990) Biochem. J. 269, p. 513-519). Cells were loaded for 1 hour at 37 ° C in loading buffer (storage buffer with 0.1% (w / v) gelatin) containing 5 μΜ of fluo-3 AM ester, washed with loading buffer and then they were resuspended in Tyrode's salt solution supplemented with 5.7 mM glucose, 0.1% (w / v) bovine serum albumin (BSA), 1.8 mM CaCl2 and 1 mM MgCl2. Cells were pipetted into black-walled, clear-bottom 96-well microplates (Costar, Boston, USA), and centrifuged (200g, 5 minutes, room temperature).

A compound of formula (I) according to the Examples was predissolved in DMSO and added to a final concentration of 0.1% (v / v) of DMSO. The tests were started by adding a concentration A₅₀ of GROa, and the transient increase in fluorescence of fluo-3 (λ_Εχ = 490 nm and λ_Ειη= 520 nm) using a FLIPR (Fluorometric Imaging Plate Reader, Molecular Devices, Sunnyvale, USA).

Compounds of formula (I) were tested according to the Examples, and found to be antagonists of the CXCR2 receptor on human neutrophils.