Thiazolo[4,5-d]pyrimidine compounds for the treatment of rheumatoid arthritis

ES 2 298 451 T3

DESCRIPTION

Thiazolo [4,5-d] pyrimidine compounds for the treatment of rheumatoid arthritis.

The present invention relates to certain thiazolopyrimidinone compounds for use in the treatment of rheumatoid arthritis.

WO 98/08847 and EP 0778277 both describe a series of 6,5-hetero-bicyclic compounds that are said to be useful as CRF antagonists.

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 four cysteine ​​motif. The chemokine superfamily can be divided into two main groups that exhibit characteristic structural motifs, the Cys-X-Cys (CXC) and Cys-Cys (CC) families. These are distinguished on the basis of the insertion of a single amino acid between the NH-proximal pair of cysteine ​​residues and sequence similarity.

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 from monocytes and lymphocytes, but not from neutrophils such as human monocyte chemotactic proteins 1-3 (MCP-1, MCP-2, and MCP-3), RANTES (Activation-Regulated, Expressed, and Secreted by Normal T Cells), eotaxin and the inflammatory proteins 1α and 1β of macrophages (MIP-1a and MIP-1 / Í).

Studies have shown that the actions of chemokines are mediated by the subfamilies of receptors coupled to the G protein, among which are the receptors designated CCR1, CCR2, CCR2A, CCR2B, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8 , CCR9, CCR10, CXCR1, CXCR2, CXCR3, CXCR4 and CX3CR1. These receptors represent successful targets for drug development, since agents that modulate these receptors could be useful in treating disorders and diseases such as those mentioned above.

According to the present invention, there is therefore provided a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof:

nr²r³

<img file="ES2298451T3_D0001.tif" />

in which

R¹ represents a C3-C7 carbocyclic group, Ci-C8 alkyl, C2-C6 alkenyl or C2-C6 alkynyl, each of the groups being optionally substituted with one or more substituent groups independently selected from halogen atoms, -OR⁴, -NR⁵R⁶, -CONR⁵R⁶, -COOR⁷, -NR⁸COR⁹, -MR¹⁰, -SO2R¹⁰, -SO2NR⁵R⁶, -NR⁸SO2R⁹or an aryl or heteroaryl group, both of which may be optionally substituted with one or more substituents independently selected from halogen atoms, and cyano, nitro, -OR groups⁴, -NR³R⁶, -CONR⁵R⁶, -COOR⁷, -NR⁸COR⁹, -MR¹⁰, -SO2R¹⁰, -SO2NR⁵R⁶, -NR⁸SO2R⁹, Ci-C alkyl₆ or trifluoromethyl;

R² and R³each independently represent a hydrogen atom, or a carbocyclic group C₃-C₇, C alkyl₁-C₈, alkenyl C₂-C₆ or C alkynyl₂-C₆, the last four groups being optionally substituted with one or more substituent groups independently selected from:

(a) halogen atoms, -OR⁴, -NR⁵R⁶, -CONR⁵R⁶, -COOR⁷, -NR⁸COR⁹, -MR¹⁰, -SO2R¹⁰, -SO2NR⁵R⁶, -NR⁸SO2R⁹;

(b) a 3-8 membered ring optionally containing one or more atoms selected from O, S, NR⁸ and which may in turn be optionally substituted with C alkyl₁-C₃ or halogen; or

ES 2 298 451 T3 (c) an aryl group or a heteroaryl group, each of which may be optionally substituted with one or more substituents independently selected from halogen atoms, cyano, nitro, -OR groups⁴, -NR⁵R⁶, -CONR⁵R⁶, -NR⁸COR⁹, -SO2NR⁵R⁶, -NR⁸SO2R⁹, alkyl QC₆ and trifluoromethyl;

R⁴ represents hydrogen, Ci-C alkyl₆ or a phenyl group, the last two of which may be optionally substituted with one or more substituent groups independently selected from halogen atoms, phenyl, -OR¹¹ and -NR¹²R¹³

R⁵ and R⁶ independently represent a hydrogen atom or a C1-C6 alkyl or phenyl group, the last two of which may be optionally substituted with one or more substituent groups independently selected from halogen, phenyl, -OR atoms¹⁴ and -NR¹⁵R¹⁶, -CONR¹⁵R¹⁶, -NR¹⁵COR¹⁶, -SONR¹⁵R¹⁶, -NR¹⁵SO2R¹⁶ or

R⁵ and R⁶, together with the nitrogen atom to which they are attached, form a saturated 4- to 7-membered heterocyclic ring system optionally containing an additional heteroatom selected from oxygen and nitrogen atoms, said ring system being optionally substituted with one or plus substituent groups independently selected from phenyl, -OR¹⁴, -COOR¹⁴, -NR¹⁵R¹⁶, -CONR¹⁵R¹⁶, -NR¹⁵COR¹⁶, -SONR¹⁵R¹⁶, -NR¹⁵SO2R¹⁶ or C1-C alkyl₆, optionally substituted with one or more substituents independently selected from halogen atoms and -NR groups¹⁵MR¹⁶ and -OR¹⁷;

R¹⁰ represents a hydrogen atom or a C alkyl group₁-C₆ or phenyl, the last two being optionally substituted with one or more substituent groups independently selected from halogen, phenyl, -OR atoms¹⁷ and -NR¹⁵R¹⁶; and each of R⁷, R⁸, R⁹, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶ and R¹⁷ independently represents a hydrogen atom or a C alkyl group₁-C₆, or phenyl; for use in the treatment of rheumatoid arthritis.

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 groups include 5- or 6-membered aromatic rings containing one or more heteroatoms selected from N, S, O. Examples include pyridine, pyrimidine, thiazole, oxazole, pyrazole, imidazole, and furan.

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

In formula (I) above, the group R¹ represents a C3-C7 carbocyclic group, C1-C8 alkyl, C2-C6 alkenyl or C2-C6 alkynyl, each of the groups being optionally substituted with one or more substituent groups independently selected from halogen atoms, -OR⁴, -NR⁵R⁶, -CONR⁵R⁶, -COOR⁷, -NR⁸COR⁹, -MR¹⁰, -SO2R¹⁰, -SO2NR⁵R⁶, -NR⁸SO2R⁹ or an aryl or heteroaryl group, both of which may be optionally substituted with one or more substituents independently selected from halogen atoms, or cyano, nitro, -OR groups⁴, -NR⁵R⁶, -CONR⁵R⁶, -COOR⁷, -NR⁸COR⁹, -MR¹⁰, -SO₂R¹⁰, -SO2NR⁵R⁶, -NR⁸SO2R⁹, C alkyl₁-C₆ or trifluoromethyl. 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 C alkyl groups₁-C₆, C alkoxy₁-C₆ or halogen atoms.

When R² and R³ represent a group substituted with one or more 3-8 membered rings optionally containing one or more atoms selected from O, S or NR⁸Examples of such groups include piperidine, pyrrolidine, piperazine, and morpholine.

Preferably one of R² and R³ is hydrogen and the other is C1-C8 alkyl substituted with hydroxy and one or more methyl or ethyl groups. More preferably, one of R² and R³ is hydrogen and the other is CH (CH3) CH₂OH, CH (Et) CH₂OH, C (CH₃)₂CH₂OH or CH (CH₂OH)₂. When one of R² and R³ is hydrogen and the other is CH (CH₃) CH₂OH or CH (Et) CH₂OH, the resulting compounds of formula (I) are preferably in the form of the (R) isomer.

Particularly preferred compounds for use in the invention include:

7 - [(2-Hydroxy-1,1-dimethylethyl) amino] -5 - [(phenylmethyl) -thio] -thiazolo [4,5-d] pyrimidin-2 (3H) -one, (R) -7- [[1- (Hydroxymethyl) propyl] amino] -5 - [(phenylmethyl) -thio] thiazolo [4,5-d] pyrimidin-2 (3H) -one, (R) -7 - [(2-Hydroxy- 1-methylethyl) amino] -5 - [(phenylmethyl) -thio] -thiazolo [4,5-d] pyrimidin-2 (3H) -one,

5 - [[(2,3-Difluorophenyl) methyl] thio] -7 - [(2-hydroxy-1,1-dimethylethyl) amino] thiazolo [4,5-d] pyrimidin-2 (3H) -one,

ES 2 298 451 T3

5 - [[(2,3-Difluorophenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino] thiazolo [4,5-d] pyrimidin-2 (3H) -one ,

5 - [[(2,3-Difluorophenyl) methyl] thio] -7 - [[2- (hydroxy-ethoxy) -ethyl] amino] thiazolo [4,5-d]] pyrimidin-2 (3H) -one ,

5 - [[(2,3-difluorophenyl) methyl] thio] -7 - [[2-hydroxy-1- (hydroxymethyl) ethyl] amino] thiazolo [4,5-d] pyrimidin-2 (3H) one,

7 - [(2-aminoethyl) amino] -5 - [[(2,3-difluorophenyl) methyl] -thio] -thiazolo [4,5-d] pyrimidin-2 (3H) -one,

5 - [[(2,3-difluorophenyl) methyl] thio] -7 - [(2-hydroxyethyl) -amino] thiazolo [4,5-d] pyrimidin-2 (3H) -one,

N- [2 - [[5 - [[(2,3-difluorophenyl) methyl] thio] -2,3-dihydro-2-oxothiazolo [4,5-d] pyrimidin-7-yl] amino] ethyl] methanesulfonamide , (+/-) - 5 - [[(2,3-difluorophenyl) methyl] thio] -7 - [[2- (2-hydroxy-ethoxy) -1-methylethyl] amino] thiazolo [4,5-d ] pyrimidin-2 (3H) -one,

7 - [[(1R) -2-amino-1-methylethyl] amino] -5 - [[(2,3-difluoro-phenyl) methyl] thio] thiazolo [4,5-d] pyrimidin-2 (3H) -ona,

5 - [[(2,3-difluorophenyl) methyl] thio] -7 - [[(1R) -2 - [(2-hydroxy-ethyl) amino] -1-methylethyl] amino] thiazolo [4,5-d ] -pyrimidin-2 (3H) -one,

5 - [[(2,3-difluorophenyl) methyl] thio] -7 - [[(1R) -2- (dimethylamino) -1-methylethyl] amino] thiazolo [4,5- d] pyrimidin-2 (3H) -ona,

5 - [[[4- (2-aminoethoxy) -3-chlorophenyl] methyl] thio] -7 - [[(1 R) -2-hydroxy -1-methylethyl] amino] thiazolo [4,5- d] pyrimidine -2 (3H) -one,

5 - [[3-Chloro-4-methoxyphenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino] thiazolo [4,5-d] pyrimidin-2 (3ph) one ,

5 - [[3-Chloro-2-fluorophenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino] thiazolo [4,5-d] pyrimidin-2 (3H) one ,

5 - [[(2,3-Difluorophenyl) methyl] thio] -7 - [[(3R, 4R) -4-hydroxy-pyrrolidin-3-yl] amino] -thiazolo [4,5-d] pyrimidin-2 (3H) -one,

5 - [[(2,3-Difluorophenyl) methyl] thio] -7 - [(3R) -pyrrolidin-3-ylamino] thiazolo [4,5-d] pyrimidin-2 (3H) -one,

7 - [[(1R) -2-Hydroxy-1-methylethyl] amino] -5 - [[(2-methyl-4-thiazolyl) methyl] thio] thiazolo [4,5-d] pyrimidin-2 (3H) -ona,

7 - [[2-Hydroxy-1- (hydroxymethyl) ethyl] amino] -5 - [[(2-methyl-4-thiazolyl) methyl] thio] thiazolo [4,5-d7p / rimidin-2 (3H) one ,

7 - [(2-Hydroxy-1,1-dimethylethyl) amino] -5 - [[(2-methyl-4-thiazolyl) methyl] thio] thiazolo [4,5-d] pyrimidin-2 (3fí) -one ,

7 - [(2-Hydroxy-1,1-dimethylethyl) amino] -5 - [[(2-methylphenyl) -ethyl) thio] thiazolo [4,5-d] pyrimidin-2 (3fí) -one,

5 - [(2-Furanylmethyl) thio] -7 - [[(1R) -2-hydroxy-1-methyl-ethyl] amino] thiazolo [4,5-d] pyrimidin-2 (3H) -one,

7 - [[(1R) -2-Amino-1-methylethyl] amino] -5 - [[(3-chloro-2-fluorophenyl) methyl] thio] thiazolo [4,5-d] pyrimidin-2 (3H) one, (2S) -2 - [[5 - [[(2,3-Difluorophenyl) methyl] thio] -2,3-dihydro-2-oxothiazolo [4,5-d] pyrimidin-7-yl] amino] -3-hydroxypropanamide,

7 - [[(1R) -2-hydroxy-1-methylethyl) amino] -5 - [(2-thienyl-methyl) -thio] thiazolo [4,5-d] pyrimidin-2 (3H) -one,

7 - [[(1R) -2-hydroxy-1-methylethyl] amino] -5 [[[3-methyl-4- (methylsulfonyl) phenyl] methyl] thio] thiazolo [4,5-d] pyrimidin2 (3H) -ona,

5 - [[[3-chloro-4- (trifluoromethoxy) phenyl] methyl] thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino] thiazolo [4,5-d] pyrimidin-2 (3H) -one,

5 - [[[2- fluoro-3- (trifluoromethyl) phenyl] methyl] thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino] thiazolo [4,5- d] pyrimidin-2 (3H) -one,

5 - [[(2,3-difluorophenyl) methyl] thio] -7- [2 - [(dimethylamino) -ethyl] amino] thiazolo [4,5-d] pyrimidin-2 (3H) -one,

ES 2 298 451 T3

5 - [[(2-fluorophenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methylethyl) amino] thiazolo [4,5-d] pyrimidin-2 (3H) -one,

7 - [[(1R) -2-hydroxy-1-methylethyl] amino] -5 - [[(2-methoxy-phenyl) -methyl] thio] thiazolo [4,5-d] pyrimidin-2 (3H) - ona,

7 - [[(1R) -2-hydroxy-1-methylethyl] amino] -5 - [(2-phenoxy-ethyl) -thio] thiazolo [4,5-d] pyrimidin-2 (3H) -one,

7 - [[(1R) -2-hydroxy-1-methylethyl] amino] -5 - [[(3-methyl-phenyl) -methyl] thio] thiazolo [4,5-d] pyrimidin-2 (3H) - ona,

5 - [[(2-fluoro-3-methylphenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino] thiazolo [4,5-d] pyrimidin-2 (3H) ona,

5 - [[(3-chlorophenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methyl-ethyl] amino] thiazolo [4,5-d] pyrimidin-2 (3H) -one ,

5 - [[(3-bromophenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methyl-ethyl] amino] thiazolo [4,5-d] pyrimidin-2 (3H) -one ,

5 - [[[4- (Diflupromentoxy) phenyl] methyl] thio] -7 - [[(1R) -2-hydroxy -1-methylethyl] amino] thiazolo [4,5- d] pyrimidin-2 (3H) - one, (+/-) - 5 - [[(2,3-difluorophenyl) methyl) thio] -7 - [[2-hydroxy-1- (methoxymethyl) ethyl] amino] thiazolo [4,5-d] pyrimidine -2 (3H) -one,

7 - [[2-hydroxy-1- (hydroxymethyl) ethyl] amino] -5 - [(phenyl-methyl) thio] thiazolo [4,5-d] pyrimidin-2 (3H) -one,

5 - [[(2-bromophenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino] thiazolo [4,5-d] pyrimidin-2 (3H) -one,

5 - [[(2,3-Difluorophenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino] thiazolo [4,5-d] pyrimidin-2 (3H) -one ,

5 - [[3-Chloro-2-fluorophenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino] thiazolo [4,5-d] pyrimidin-2 (3H) one , (+/-) - 5 - [[(2,3-difluorophenyl) methyl] thio] -7 - [[2-hydroxy-1- (methoxymethyl) ethyl] amino] thiazolo [4,5-d] pyrimidine- 2 (3H) -one,

7 - [[2-hydroxy-1- (hydroxymethyl) ethyl] amino] -5 - [(phenyl-methyl) thio] thiazolo [4,5-d] pyrimidin-2 (3H) -one,

7 - [[(1R) -2-Hydroxy-1-methylethyl] amino] -5 - [(phenylmethyl) -thio] thiazolo [4,5-d] pyrimidin-2 (3H) -one,

5 - [(5-chloro-1,2,3-thiadiazol-4-yl) thio] -7 - [[(1R) -hydroxy-1-methylethyl] amino] -thiazolo [4,5-d] pyrimidin- 2 (3H) one, and its pharmaceutically acceptable salts and solvates.

Particular salts of compounds of formula (I) include:

5 - [[(2,3-Difluorophenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino] thiazolo [4,5-d] pyrimidin-2 (3H) -one , sodium salt,

5 - [[3-Chloro-2-fluorophenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino] thiazolo [4,5-d] pyrimidin-2 (3H) one , sodium salt, (+/-) - 5 - [[(2,3-difluorophenyl) methyl] thio] -7 - [[2-hydroxy-1- (methoxymethyl) ethyl] amino] thiazolo [4,5- d] pyrimidin-2 (3H) -one, sodium salt

7 - [[2-hydroxy-1- (hydroxymethyl) ethyl] amino] -5 - [(phenyl-methyl) thio] thiazolo [4,5-d] pyrimidin-2 (3fí) -one, sodium salt, or

7 - [[(1R) -2-Hydroxy-1-methylethyl] amino] -5 - [(phenyl-methyl) thio] thiazolo [4,5-d] pyrimidin-2 (3fí) -one, sodium salt.

Additional particular salts of compounds of formula (I) include:

7 - [[(1R) -2-amino-1-methylethyl] amino] -5 - [[(2,3-difluorophenyl) methyl] thio] thiazolo [4,5-d] pyrimidin-2 (3H) -one , trifluoroacetate,

5 - [[(2,3-difluorophenyl) methyl] thio] -7 - [[(1R) -2 - [(2-hydroxyethyl) amino] -1-methylethyl] amino] thiazolo [4,5-d] pyrimidine -2 (3H) -one, trifluoroacetate,

5 - [[(2,3-Difluorophenyl) -methyl] thio] -7 - [[(1R) -2- (dimethylamino) -1-methylethyl] amino] -thiazolo [4,5-d] pyrimidin-2 ( 3H) -one,

ES 2 298 451 T3

5 - [[[4- (2-aminoethoxy) -3-chlorophenyl] methyl] thio] -7 - [[(1R) -2-hydroxy -1-methylethyl] amino] thiazolo [4,5-d] pyrimidine- 2 (3H) -one, trifluoroacetate,

5 - [[(2,3-difluorophenyl) methyl] thio] -7- [2 - [(dimethylamino) -ethyl] amino] thiazolo [4,5-d] pyrimidin-2 (3H) -one, monohydrochloride, or

5 - [[(2,3-Difluorophenyl) methyl] thio] -7 - [(3R) -pyrrolidin-3-ylamino] thiazolo [4,5-d] pyrimidin-2 (3ph) -one, dihydrochloride.

Also provided is a process for the preparation of a compound of formula (I) comprising either: Treatment of a compound of formula (IIA)

<img file="ES2298451T3_D0002.tif" />

where R¹, R² and R³ are as defined in formula (I) with a thiol R¹ SH in the presence of a suitable base and optional formation of a pharmaceutically acceptable salt. The reaction can be carried out in a mixed solvent of DMSO and ethanol at a temperature between 0 ° C and 100 ° C using sodium borohydride as the base.

Compounds of formula (IIA) where R¹, R² and R³ are as defined in formula (I) can be prepared by treating compounds of formula (I) with a suitable oxidizing agent such as oxone. The reaction can be carried out in a solvent such as acetonitrile at a temperature between 0 ° C and 100 ° C.

Or, treatment of a compound of formula (IIB):

NR * R³

<img file="ES2298451T3_D0003.tif" />

(UB) where R¹, R² and R³ are as defined in formula (I) and X is a leaving group with a metal alkoxide, followed by treatment with an acid or base and optional formation of a pharmaceutically acceptable salt.

X is any suitable leaving group such as halogen. The reaction can be carried out in an alcohol solvent such as methanol and the deprotection is carried out in a solvent such as 1,4-dioxane. Examples of metal alkoxides include potassium methoxide. Examples of suitable acids include hydrochloric acid. Preferably, the compound of formula (IIB) is treated with a metal alkoxide such as potassium methoxide followed by an acid such as concentrated HCl in a solvent such as 1,4-dioxane.

Compounds of formula (IIB) where R¹, R² and R³ are as defined in formula (I) and X is a halogen, they can be prepared from corresponding compounds (IIB) where R¹, R² and R³ are as defined in formula (I) and X is NH₂ by treatment with a diazotizing agent such as isoamyl nitrite and a halogenating agent such as bromoform.

ES 2 298 451 T3

Compounds of formula (IIB) where R¹, R² and R³ are as defined in formula (I) and X is NH₂ can be prepared by treating a compound of formula (IIIA):

NR * R³

<img file="ES2298451T3_D0004.tif" />

(ΠΙΑ) where R² and R³ are as defined in formula (I) and X is NH₂ 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 diisopropylethylamine in an inert solvent such as DMSO / N-methylpyrrolidinone at a temperature between 0 ° C and 100 ° C.

Compounds of formula (IIIA) where R² and R³ are as defined in formula (I) and X is NH2 can be prepared by treating a compound of formula (IIB) where R² and R³ are as defined in formula (I), X is NH₂ and R¹ is a suitable benzyl group such as benzyl or 2,3-difluorobenzyl with a reducing medium such as metallic sodium in liquid ammonia, or by treatment of a compound of formula (IIIB):

<img file="ES2298451T3_D0005.tif" />

where R¹ is as defined in formula (I) and L is a leaving group such as chlorine with an amine HNR²R³ where R² and R³ are as defined in formula (I). The reaction can be carried out in a solvent such as N-methylpyrrolidine at a temperature between 0 ° C and 150 ° C. Compounds of formula (IIIB) where R¹ is as defined in formula (I) and L is a halogen can be prepared by treating a compound of formula (IIIB) where R¹is as defined is formula (I) and L is a hydroxyl group with a halogenating agent such as phosphorous oxychloride.

The reaction can be carried out in the presence of refluxing dimethylaniline.

Compounds of formula (IIIB) where R¹ is as defined in formula (I) and L is a hydroxyl group can be formed by treating a compound of formula (IVA) 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 potassium eerc-butoxide in an inert solvent such as DMSO at room temperature.

<img file="ES2298451T3_D0006.tif" />

ES 2 298 451 T3

Or by heating a compound of formula (IVB) where R¹ it is as defined above.

<img file="ES2298451T3_D0007.tif" />

The reaction is preferably carried out in a suitable solvent such as DMF at elevated temperature, for example about 120 ° C.

Compounds of formula (IVB) can be easily prepared by reacting a compound of general formula (V) where R¹ it is as defined above, with bromine potassium thiocyanate in an inert solvent such as dimethylformamide / pyridine.

<img file="ES2298451T3_D0008.tif" />

Compounds of formula (V) are conveniently prepared by reacting a compound of formula (VI):

Oh

<img file="ES2298451T3_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 hydride in an inert solvent such as DMF at room temperature.

Compounds of formula (IVA) and (VI) are either commercially available or well known in the literature.

It will be appreciated by those skilled in the art that in the processes described above the functional groups (eg hydroxyl groups) of the intermediates may need to be protected by protecting groups. The final step in the preparation of the compounds of the invention may involve the removal of one or more protecting groups. The protection and deprotection of functional groups is described in detail in "Protective Groups in Organic Chemistry", compiled by JWF McOmic. Plenum Press (1973), and "Protective Groups in Organic Synthesis", 2^toedition, TW Greene &amp; PGM Wuts, Wiley-Interscience (1991).

The compounds of formula (1) above can be converted into a pharmaceutically acceptable salt or solvate thereof, preferably a base addition salt such as sodium, potassium, calcium, aluminum, lithium, magnesium, zinc, benzathine, 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.

ES 2 298 451 T3

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

(1) (the respiratory tract) obstructive airway diseases including chronic obstructive pulmonary disease (COPD); asthma, such as bronchial, allergic, intrinsic, extrinsic and dust-caused asthma, particularly chronic or inveterate asthma (eg late asthma and airway hyper-sensitivity); 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 scrofulous rhinitis; seasonal rhinitis, including nervous rhinitis (hay fever) and vasomotor rhinitis; sarcoidosis; farmer's lung and related diseases, fibroid lung and idiopathic interstitial pneumonia;

(2) (bones 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, bullous pemphigus, bullous epidermolysis, urticaria, angioderma, vasculitis, erythema, cutaneous eosinophilia, uveitis, and alopeitis spring;

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

(5) (other tissues and systemic diseases) multiple sclerosis, atherosclerosis, Acquired Immune Deficiency Syndrome (AIDS), lupus erythematosus; systemic lupus, erythematosus, Hashimoto's thyroiditis, myasthenia gravis, type I diabetes, nephrotic syndrome, eosinophilia fasciitis, hyper IgE syndrome, lepromatous leprosy, Sezary syndrome, and idiopathic thrombocytopenic purpura, postoperative adhesions, and septicemia;

(6) acute and chronic (allograft rejection) after, for example, kidney, heart, liver, lung, bone marrow, skin and cornea transplantation; and reverse rejection disease;

(7) cancers, especially non-small cell lung cancer (NSCLC), malignant melanoma, prostate cancer, and squamous sarcoma, as well as tumor metastases;

(8) diseases in which angiogenesis is associated with elevated levels of the chemokine CXCR2 (eg NSCLC, diabetic retinopathy);

(9) cystic fibrosis, stroke, reperfusion injury to the heart, brain, extremities, and other organs;

(10) burn wounds and chronic skin ulcers;

(11) reproductive diseases (eg ovulation, menstruation and implantation disorders, premature labor, 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 the treatment of rheumatoid arthritis.

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 being the CXCR2 receptor.

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 the treatment of rheumatoid arthritis.

In yet another 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 the treatment of arthritis. rheumatoid.

In the context of the present specification, the term "therapy" also includes "prophylaxis" unless specific indications are made to the contrary. The terms "therapeutically" and "therapeutically" should be interpreted in the same sense.

ES 2 298 451 T3

For the above-mentioned therapeutic uses, the dose administered will, of course, vary with the compound employed, the mode of administration, the treatment desired, and the condition in question.

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 / salt / solvate of formula (I) (ingredient active) 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% w (weight percent), more preferably from 0.05 to 80% w, still more preferably from 0.10 to 70% w, and still more preferably from 0.10 to 50 wt% 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 conditions can be administered topically (eg to the lung and / or airways or skin) in the form of solutions, suspensions, aerosols with heptafluoroalkanes and dry powder formulations; or systemically, eg by administration or 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 by transdermal route. Preferably, the compounds of the invention are administered via or al.

The invention will be further illustrated below with reference to the examples that follow. 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. If necessary, reactions were carried out under an inert nitrogen or argon atmosphere. Chromatography was generally performed using Silica 60® Matrex (35-70 microns) or Silica Gel 60® Prolabo (35-70 microns) suitable for silica gel flash chromatography. High pressure liquid chromatography purification was performed using a Waters Micromass LCZ equipped with a Waters 600 pump controller, Waters 2487 detector and Gilson FC024 or Waters Delta Prep 4000 fraction collector. Abbreviations pf and DMSO used in the examples mean melting point and dimethyl sulfoxide, respectively.

Example 1

7 - [(2-Hydroxy-1,1-dimethylethyl) amino] -5 - [(femlmethyl) thio] -thiazolo [4,5-d] pyrimidin-2 (3H) -one (a) Thiocyanic acid, 6- amino-1,4-dihydro-4-oxo-2 - [(phenylmethyl) thio] -5-pyrimidinyl-ester

6-amino-2 - [(phenylmethyl) thio] 4 (1H) -pyrimidinone (10.5 g) [preparation as described in WO 9635678] and potassium thiocyanate (25 g) were heated together at 65 ° C 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 510 ° C. The reaction mixture was poured into ice water, stirred for one 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) 2-Amino-5 - [(phenylmethyl) thio] thiazolo [4,5-d] pyrimidin-7 (4H) -one

The product from Example 1, step a) (7.35 g) was heated at 120 ° C in N, N-dimethylformamide (40 ml) / water (10 ml) for 10 hours. After cooling, the resulting solid was filtered off, washed with water, and then with ethyl acetate to give the subtitle compound.

mp 325 ° C

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

(c) 7-Chloro-5 - [(phenylmethyl) thio] thiazolo [4,5-d] pyrimidin-2-amine

The product from Example 1, step b) (0.89 g), phosphorous oxychloride (12 ml) and N, N-dimethylaniline (1.2 ml) were heated under reflux for 2 hours. The cooled reaction mixture was poured into ice water and stirred for 2 hours. Chromatography (SiO₂, methanol / dichloromethane as eluent) gave the subtitle compound.

ES 2 298 451 T3

mp 217-218.5 ° C

MS (APCI) 309 (M + H, 100%).

(d) 2 - [[2-Amino-5 - [(femlmethyl) thio] thiazolo [4,5-d] pyrimidin-7-yl] amino] -2-methyl-1-propan.ol

The product from Example 1, step c) (0.6 g), and 1-amino-2-methyl-propan-2-ol (1.1 g) in tetrahydrofuran (10 ml) was heated in a hermetically sealed container to 100 ° C for 18 hours. The mixture was evaporated to dryness and purified (SiO₂, ethyl acetate as eluent) to give the subtitle compound (0.46 g).

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

(e) 2 - [[2-Bromo-5 - [(femlmethyl) thio] thiazolo [4,5-d] pyrimidin-7-yl] amino] -2-methyl-1-propanol

To a solution of the product from Example 1, step d) (0.1 g) in bromoform (5 ml) was added isoamyl nitrite (0.13 ml) and the mixture was heated at 60 ° C for 10 min. The mixture was evaporated to dryness and purified (SiO₂, ethyl acetate: dichloromethane 1: 9 as eluent) to give the subtitle compound as a colorless solid (0.43 g).

MS (APCI) 427 (M + H, 100%).

(f) 2 - [[2-Methoxy-5 - [(femlmethyl) thio] thiazolo [4,5-d] pyrimidin-7-yl] amino] -2-methyl-1-propan.ol

To a solution of the product from Example 1, step e) (0.36 g) in methanol (5 ml) was added potassium hydroxide (0.095 g) and the mixture was stirred for 30 min. The mixture was neutralized with concentrated hydrochloric acid, then evaporated to dryness and purified (SiO₂, ethyl acetate: dichloromethane 1: 9 as eluent) to give the subtitle compound as a colorless solid (0.245 g).

MS (APCI) 377 (M + H, 100%).

(g) 7 - [(2-hydroxy-1,1-dimethylethyl) amino] -5 - [(phenylmethyl) -thio] thiazolo [4,5-d] pyrimidin-2- (3H) -one

To a solution of the product from Example 1, step f) (0.21 g) in 1,4-dioxane (5 ml) were added water (0.1 ml) and concentrated hydrochloric acid (one drop). The mixture was heated at 45 ° C for 3 hours, and then evaporated to dryness. Recrystallization (acetonitrile) gave the title compound (0.110 g).

mp 207-8 ° C

MS (APCI) 363 (M + H⁺, 100%).

NMR dH (d₆-DMSO) 12.37 (1H, s), 7.43-7.23 (5H, m), 6.61 (1H, bs), 4.81 (1H, t), 4.34 (2H, s ), 3.55 (2H, bs), 1.32 (6H, s).

Example 2 (R) -7 - [[1- (Hydroxymethyl) propyl] amino] -5 - [(phenylmethyl) -thio] thiazolo) [4,5-d) pyrimidin-2- (3H) -one (a) (R) -2 - [[2-Amino-5 - [(phenylmethyl) thio] thiazolo [4,5-d] -pyrimidin-7-yl] amino] -1-butanol

To a mixture of the product of Example 1 step c) (2.5 g) and (R) - (-) - 2-amino-1-butanol (5 g) in a solvent of N-methylpyrrolidinone (10 ml) was added N, N-diisopropyl-ethylamine (5 ml) and the resulting mixture was heated at 100 ° C for 10 hours. The mixture was poured into water and the product was collected by filtration to give the subtitle compound (2.5 g).

MS (APCI) 362 (M + H⁺, 100%).

(b) (R) -2 - [[2-Bromo-5 - [(phenylmethyl) thio] thiazolo [4,5-d] -pyrimidin-7-yl] amino] -1-butanol

Prepared by the method of Example 1 step e), using the product of Example 2 step a).

MS (APCI) 427 (M + H⁺, 100%).

ES 2 298 451 T3 (c) (R) -2 - [[2-Methoxy-5 - [(phenylmethyl) thio] thiazolo [4,5-d] -pyrimidin-7-yl] amino] -1-butanol

Prepared by the method of Example 1 step f), using the product of Example 2 step b).

MS (APCI) 377 (M + H⁺, 100%).

(d) (R) -7 - [[1- (Hydroxymethyl) propyl] amino] -5 - [(phenyl-methyl) thio] thiazolo [4,5-d] pyrimidin-2 (3H) -one

Prepared by the method of Example 1 step g), using the product of Example 2 step c).

MP 217-8 ° C

MS (APCI) 363 (M + H⁺, 100%).

NMR OH (d6-DMSO) 12.37 (1H, s), 7.43-7.21 (6H, m), 4.68 (1H, t), 4.32 (2H, q), 4.09 (1H, bs), 3.47-3.32 (2H, m), 1.69-1.59 (1H, m), 1.48-1.41 (1H, m), 0.82 (3H , t).

Example 3 (R) -7 - [(2-Hydroxy-1-methylethyl) amino] -5 - [(phenylmethyl) -thio] thiazolo [4,5-d] pyrimidin-2 (3H) -one (a) ( R) -2 - [[2-Amino-5 - [(phenylmethyl) thio] thiazolo [4,9-d] -pyrimidin-7-yl] amino] -1-propanol

Prepared by the method of Example 2 step a), using the product of Example 1 step c) and (R) - (-) - 2-amino-1-propanol.

MS (APCI) 412 (M + H⁺, 100%).

(b) (R) -2 - [[2-Bromo-5 - [(phenylmethyl) thio] thiazolo [4,5-d] -pyrimidin-7-yl] amino] -1-propanol

Prepared by the method of Example 1 step b), using the product of Example 3 step a).

MS (APCI) 348 (M + H⁺, 100%).

(c) (R) -2 - [[2-Methoxy-5 - [(phenylmethyl) thio] thiazolo [4,5-d] -pyrimidin-7-yl] amino] -1-propanol

Prepared by the method of Example 1 step f), using the product of Example 3 step b).

MS (APCI) 363 (M + H⁺, 100%).

(d) (R) -7 - [[2-Hydroxy-1-methylethyl] amino] -5 - [(phenylmethyl) -thio] thiazolo [4,5-d] pyrimidin-2 (3H) -one

Prepared by the method of Example 1 step g), using the product of Example 3 step c).

MS (APCI) 349 (M + H⁺, 100%).

NMR OH (d₆-DMSO) 12.38 (1H, s), 7.44-7.20 (6H, m), 4.72 (1H, t), 4.32 (2H, m), 4.23 (1H, m ), 3.49-3.29 (2H, m), 1.11 (3H, d).

Example 4

5 - [[(2,3-Difluorophenyl) methyl] thio] -7 - [(2-hydroxy-1,1-dimethylethyl) amino] thiazolo [4,5-d] pyrimidin-2 (3H) -one (a ) 2-Amino-5 - [[(2,3-difluorofeml) methyl] thio] thiazolo [4,5-dJpyrimidin-7 (4H) -one

Potassium i-butoxide solution (0.45 ml of 1M solution in tetrahydrofuran) was added to a stirred solution of 2-amino-5,6-dihydro-5-thioxo-thiazolo [4,5-d] pyrimidin-7 (4H) -one (0.09 g) [cited in: Indian J. Chem., Sect. B (1989), 28 B (11), 964-5.] And 2,3-difluorobenzyl bromide in dimethyl- sulfoxide (2 ml). After stirring for 3 days, the reaction mixture was poured into water to give the subtitle compound, which was isolated by filtration.

MS (APCI) 327 (M + H⁺, 100%).

ES 2 298 451 T3 (b) 7-Chloro-5 - [[(2,3-difluorophenyl) methyl] thio] thiazolo [4,5-dJpyrimidin-2-amine

Prepared by the method of Example 1 step c), using the product of Example 4 step a).

MS (APCI) 345 (M + H⁺, 100%).

(c) 2 - [[2-Amino-5 - [[(2,3-difluorophenyl) methylJthioJ-thiazolo [4,5-dJpyrimidin-7-ylJaminoJ-2-methyl-1-propanol

Prepared by the method of Example 2 step a), using the product of Example 4 step b) and 2-amino-2-methylpropanol.

MS (APCI) 398 (M + H⁺, 100%).

(d) 2 - [[2-Bromo-5 - [[(2,3-difluorophenyl) methylJthioJthiazolo [4,5-dJpyrimidin-7-ylJaminoJ-2-methyl-1-propanol

Prepared by the method of Example 1 step e), using the product of Example 4 step c).

MS (APCI) 462 (M + H⁺, 100%).

(e) 2 - [[5 - [[(2,3-Difluorophenyl) methylJthioJ-2-methoxythiazolo [4,5-dJpyrimidin-7-ylJaminoJ-2-methyl-1-propanol

Prepared by the method of Example 1 step f), using the product of Example 4 step d).

MS (APCI) 413 (M + H⁺, 100%).

(f) 5 - [[(2,3-Difluorophenyl) methylJthioJ-7- (2-hydroxy-1,1-dimethylethyl) aminoJthiazolo [4,5-dJpyrimidin-2 (3H) -one

Prepared by the method of Example 1 step f), using the product of Example 4 step e).

MS (APCI) 399 (M + H⁺, 100%).

NMR OH (d₆-DMSO) 12.41 (1H, s), 7.41-7.30 (2H, m), 7.21-7.13 (1H, m), 6.64 (1H, bs), 4.79 (1H, t), 4.41 (2H, s), 3.53 (2H, d), 1.29 (6H, s).

Example 5

5 - [[(2,3-Difluorophenyl) methylJthioJ-7 - [[(1R) -2-hydroxy-1-methylethylJaminoJthiazolo [4,5-dJpyrimidyl-2- (3H) -one (a) (2R) -2 - [[2-amino-5 - [[(2,3-difluorophenyl) methylJthioJ-thiazolo [4,5-dJpyrimidin-7-ylJaminoJ-1-propanol

Prepared by the method of Example 2 step a), using the product of Example 4 step b) and (R) - (-) - 2-amino-1-propanol.

MS (APCI) 384 (M + H⁺, 100%).

(b) (2R) -2 - [[2-Bromo-5 - [[(2,3-difluorophenyl) methylJthioJ-thiazolo [4,5-dJpyrimidin-7-ylJaminoJ-1-propanol

Prepared by the method of Example 1 step e), using the product of Example 5 step a).

MS (APCI) 448 (M + H⁺, 100%).

(c) (2R) -2 - [[5 - [[(2,3-Difluorophenyl) methylJthioJ-2-methoxy-thiazolo [4,5-dJpyrimidin-7-ylJaminoJ-1-propanol

Prepared by the method of Example 1 step f), using the product of Example 5 step b).

MS (APCI) 398 (M + H⁺, 100%).

(d) 5 - [[(2,3-Difluorophenyl) methylJthioJ-7 - [[(1R) -2-hydroxy-1-methylethylJaminoJ-thiazolo [4,5-dJpyrimidin-2 (3H) -one

Prepared by the method of Example 1 step g), using the product of Example 5 step c).

ES 2 298 451 T3

MS (APCI) 385 (M + H⁺, 100%).

NMR δΗ (d₆-DMSO) 12.41 (1H, s), 7.41-7.11 (4H, m), 4.72 (1H, t), 4.39 (2H, m), 4.21 (1H, m ), 3.47-3.29 (2H, m), 1.09 (3H, d).

Example 6

5 - [[(2,3-Difluorophenyl) meM] thio] -7 - [[2- (hidn) xethoxy) -ethyl] amino] thiazolo [4,5-d] pyrimidin-2- (3H) -one ( a) 2- [2 - [[2-Amino-5 - [[(2,3-difluorophenyl) methyl] thio] thiazolo [4,5-d] pyrimidin-7-yl] amino] ethoxy] ethanol

Prepared by the method of Example 2 step a), using the product of Example 4 step b) and 2- (2-aminoethoxy) ethanol.

MS (APCI) 414 (M + H⁺, 100%).

(b) 2- [2 - [[2-Bromo-5 - [[(2,3-difluorophenyl) methyl] thio] -thiazolo [4,5-d] pyrimidin-7-yl] amino] ethoxy] ethanol

Prepared by the method of Example 1 step e), using the product of Example 6 step a).

MS (APCI) 478 (M + H⁺, 100%).

(c) 2- [2 - [[5 - [[(2,3-Difluorofeml) methyl] thio] -2-methoxythiazolo [4,5-d] pyrimidin-7-yl] amino] ethoxy] ethanol

Prepared by the method of Example 1 step f), using the product of Example 6 step b).

MS (APCI) 429 (M + H⁺, 100%).

(d) 5 - [[(2,3-Difluorophenyl) methyl] thio] -7 - [[2- (2-hydroxy-ethoxy) ethyl] amino] -thiazolo [4,5-d] pyrimidin-2 (3H ) -ona

Prepared by the method of Example 1 step g), using the product of Example 6 step c).

MP 213-4 ° C

MS (APCI) 415 (M + H⁺, 100%).

NMR δΗ (d6-DMSO) 12.41 (1H, s), 7.39-7.11 (4H, m), 4.57 (1H, t), 4.39 (2H, s), 3.57 -3.38 (8H, m).

Example 7

5 - [[(2,3-Difluorophenyl) methyl] thio] -7 - [[2-hydroxy-1- (hydroxymethyl) ethyl] amino] thiazolo [4,5-d] pyrimidin-2- (3H) -one (a) 2 - [[2-Amino-5 - [[(2,3-difluorophenyl) methyl] thio] thiazolo- [4,5-d] pyrimidin-7-yl] amino] -1,3-propanediol

Prepared by the method of Example 2 step a), using the product of Example 4 step b) and 2-amino-1,3-propanediol.

MS (APCI) 400 (M + H⁺, 100%).

(b) 2 - [[2-Bromo-5 - [[(2,3-difluorophenyl) methyl] thio] thiazolo- [4,5-d] pyrimidin-7-yl] amino] -1,3-propanediol

Prepared by the method of Example 1 step e), using the product of Example 7 step a).

MS (APCI) 464 (M + H⁺, 100%).

(c) 2 - [[5 - [[(2,3-Difluorophenyl) methyl] thio] -2-methoxythiazolo [4,5-d] pyrimidin-7-yl] amino] -1,3-propanediol

Prepared by the method of Example 1 step f), using the product of Example 7 step b).

MS (APCI) 415 (M + H⁺, 100%).

ES 2 298 451 T3 (d) 5 - [[(2,3-Difluorophenyl) methyl] thio] -7 - [[2-hydroxy-1- (hydroxymethyl) ethyl] amino] thiazolo [4,5-d] pyrimidine -2 (3H) -one

Prepared by the method of Example 1 step g), using the product of Example 7 step c).

MP 178-9 ° C

MS (APCI) 401 (M + H⁺, 100%).

NMR δΗ (d₆-DMSO) 12.41 (1H, s), 7.42-7.11 (4H, m), 4.66 (2H, s), 4.40 (2H, s), 4.19 (1H, m ), 3.49 (4H, m).

Example 8

7 - [(2-Aminoethyl) amino] -5 - [[(2,3-difluorophenyl) methyl] thio] -thiazolo [4,5-d] pyrimidin-2- (3H) -one (a) Acid 2- [[2-amino-5 - [[(2,3-difluorophenyl) methyl] thio] -thiazolo [4,5-dJ-pyrimidin-7-yl] amino] ethyl) -carbamic ester

1,1-dimethylethyl

Prepared by the method of Example 2 step a), using the product of Example 4 step b) and (2-aminoethyl) carbamic acid, 1,1-dimethylethyl ester.

MS (APCI) 469 (M + H⁺, 100%).

(b) [2 - [[2-Bromo-5 - [[(2,3-difluorophenyl) methyl] thio] -thiazolo [4,5-dJ-pyrimidin-7-yl] amino] ethyl) -carbamic acid, ester

1,1-dimethylethyl

Prepared by the method of Example 1 step e), using the product of Example 8 step a).

MS (APCI) 533 (M + H⁺, 100%).

(c) [2 - [[5 - [[(2,3-Difluorophenyl) methyl] thio] -2-methoxythiazolo [4,5-d] pyrimidin-7-yl] amino] ethyl) -carbamic acid, ester

1,1-dimethylethyl

Prepared by the method of Example 1 step f), using the product of Example 8 step b).

MS (APCI) 489 (M + H⁺, 100%).

(d) 7 - [(2-Aminoethyl) amino] -5 - [[(2,3-difluorophenyl) methyl] -thio] thiazolo [4,5-dJpyrimidin-2 (3H) -one

Prepared by the method of Example 1 step g), using the product of Example 8 step c).

MP 215-6 ° C

MS (APCI) 370 (M + H⁺, 100%).

NMR δΗ (d6-DMSO) 12.00 (1H, s), 7.45-7.11 (3H, m), 6.35 (1H, bs), 4.37 (2H, s), 3.48 (2H, m), 2.92 (2H, t).

Example 9

5 - [[(2,3-Difluorophenyl) methyl] thio] -7 - [(2-hydroxyethyl) -amino] thiazolo [4,5-dJ-pyrimidin-2- (3H) -one (a) 2 - [[2 -Amino-5 - [[(2,3-difluorophenyl) methyl] thio] thiazolo- [4,5-dJpyrimidin-7-yl] amino] ethanol

Prepared by the method of Example 2 step a), using the product of Example 4 step b) and ethanolamine.

MS (APCI) 370 (M + H⁺, 100%).

(b) 2 - [[2-Bromo-5 - [[(2,3-difluorophenyl) methyl] thio] thiazolo [4,5-dJpyrimidin-7-yl] amino] ethanol

Prepared by the method of Example 1 step e), using the product of Example 9 step a).

MS (APCI) 434 (M + H⁺, 100%).

ES 2 298 451 T3 (c) 2 - [[5 - [[(2,3-D (fluorq / enzy7) meíz77ízo7-2-meíoxzízazo / or [4,5-d7p / r / mzd / n-7-z // am / no7eian /

Prepared by the method of Example 1 step f), using the product of Example 9 step b).

MS (APCI) 385 (M + H⁺, 100%).

(d) 5 - [[(2,3-Difluorophenyl) methyl] thio] -7 - [(2-hydroxyethyl) -amino] thiazolo [4,5-d] pyrimidin-2 (3H) -one

Prepared by the method of Example 1 step g), using the product of Example 9 step c).

MP 217-9 ° C

MS (APCI) 371 (M + H⁺, 100%)

NMR δΗ (de-DMSO) 12.43 (1H, s), 7.67-7.64 (1H, m), 7.39-7.33 (2H, m), 7.16-7.12 ( 1H, m), 4.73 (1H, t), 4.40 (2H, s), 3.52-3.42 (4H, m).

Example 10

N- [2 - [[5 - [[(2,3-Difluorophenyl) methyl] thio] -2,3-dihydro-2-oxothiazolo [4,5-d] pyrimidin-7-yl] amino] ethyl] methanesulfonamide (a) N- [2 - [[2-Amino-5 - [[(2,3-difluorophenyl) methyl] thio] thiazolo [4,5-d] pyrimidin-7-yl] amino] ethyl] methanesulfonamide

Prepared by the method of Example 2 step a), using the product of Example 4 step b) and N- [2-aminoethyl] methanesulfonamide.

MS (APCI) 448 (M + H⁺, 100%).

(b) N- [2 - [[2-Bromo-5 - [[(2,3-difluorophenyl) methyl] thio] thiazolo [4,5-d] pyrimidin-7-yl] amino] ethyl] methanesulfonamide

Prepared by the method of Example 1 step e), using the product of Example 10 step a).

MS (APCI) 511 (M + H⁺, 100%).

(c) N- [2 - [[5 - [[(2,3-Difluorophenyl) methyl] thio] -2-methoxythiazolo [4,5-dJ-pyrimidin-7-yl] amino] ethyl] methanesulfonamide

Prepared by the method of Example 1 step f), using the product of Example 10 step b).

MS (APCI) 462 (M + H⁺, 100%).

(d) N- [2 - [[5 - [[(2,3-Difluorophenyl) methyl] thio] -2,3-dihydro-2-oxothiazolo [4,5-dJpyrimidin-7-yl] amino] ethyl] methanesulfonamide

Prepared by the method of Example 1 step g), using the product of Example 10 step c).

MP 225-6 ° C

MS (APCI) 448 (M + H⁺, 100%).

NMR δΗ (de-DMSO) 12.49 (1H, s), 7.72 (1H, t), 7.41-7.13 (4H, m), 4.43 (2H, bs), 3.49 (2H, m), 3.13 (2H, m), 2.89 (3H, s).

Example 11 (+/-) - 5 - [[(2,3-Difluorophenyl) methyl] thio] -7 - [[2- (2-hydroxy-ethoxy) -1-methylethyl] amino] thiazolo [4,5- dJpyrimidin-2 (3H) -one (a) (+/-) - 2- [2 - [[2-Amino-5 - [[(2,3-difluorophenyl) methyl] thio] -thiazolo [4,5- dJpyrimidin-7-yl] amino] propoxy] ethanol

Prepared by the method of Example 2 step a), using the product of Example 4 step b) and (±) -2- [2-aminopropoxy] ethanol.

ES 2 298 451 T3

MS (APCI) 428 (M + H⁺, 100%).

(b) (+/-) - 2- [2 - [[2-Bromo-5 - [[(2,3-difluorophenyl) methyl] thio] -thiazolo [4,5-d] pyrimidin-7-yl] amino] propoxy] ethanol

Prepared by the method of Example 1 step e), using the product of Example 11 step a).

MS (APCI) 492 (M + H⁺, 100%).

(c) (+/-) - 2- [2 - [[5 - [[(2,3-Difluorophenyl) methyl] thio] -2-methoxythiazolo [4,5-d] pyrimidin-7-yl] amino] propoxy] ethanol

Prepared by the method of Example 1 step f), using the product of Example 1 step b).

MS (APCI) 443 (M + H⁺, 100%).

(d) (±) -5 - [[(2,3-Difluorophenyl) methyl] thio] -7 - [[2- (2-hydroxyethoxy) -1-methylethyl] amino] thiazolo [4,5-d] pyrimidine -2 (3H) one

Prepared by the method of Example 1 step g), using the product of Example 11 step c).

MP 221-2 ° C

MS (APCI) 429 (M + H⁺, 100%)

NMR OH (d6-DMSO) 12.43 (1H, s), 7.47-7.30 (3H, m), 7.17-7.13 (1H, m), 4.56 (1H, t) , 4.40 (2H, s), 4.35 (1H, m), 3.49-3.32 (6H, m), 1.10 (3H, d).

Example 12

7 - [[(1R) -2-Amino-1-methylethyl] amino] -5 - [[(2,3-difluorophenyl) methyl] thio] thiazolo [4,5-d] pyrimidin-2 (3H) -one , trifluoroacetate (a) (2R) -2 - [[2-Amino-5 - [[(2,3-difluorophenyl) methyl] thio] -thiazolo [4,5-d] pyrimidin-7-yl] amino] propanamide

Prepared by the method of Example 2 step a), using the product of Example 4 step b) and (2R) -2-amino-propanamide hydrochloride.

MS (APCI) 397 (M + H⁺, 100%).

(b) N '- [(1R) -2-Amino-1-methylethyl] -5 - [[(2,3-difluorofeml) methyl] thio] thiazolo [4,5-dJpyrimidine-2,7-diamine

To a solution of the product of Example 12 step a) (0.3 g) in dry tetrahydrofuran (10 ml) was added 2M borane in THF (10 ml) and the mixture was heated under reflux for 6 hours. It was quenched while hot with methanol (30 ml), evaporated to dryness and the residue was taken up in methanol (30 ml) containing a few drops of concentrated hydrochloric acid. The mixture was then refluxed for an additional hour, and evaporated to dryness to give a pale yellow solid.

MS (APCI) 383 (M + H⁺, 100%).

(c) [(2R) -2 - [[2-amino-5 - [[(2,3-difluorophenyl) -methyl] thio] thiazolo [4,5-d] pyrimidin-7-yl] amino] propyl acid ] -carbamic, 1,1-dimethylethyl ester

To a solution of the product from Example 12 step b) (1.6 g) in THF (50 ml) was added ditert-butyl bicarbonate (0.91 g) and the mixture was stirred for 2 days. It was evaporated to dryness to give 2.0 g.

MS (APCI) 483 (M + H⁺, 100%).

(d) [(2R) -2 - [[2-bromo-5 - [[(2,3-difluorophenyl) -methyl] thio] thiazolo [4,5-d] pyrimidin-7-yl] amino] propyl acid ] -carbamic, 1,1-dimethylethyl ester

Prepared by the method of Example 1 step e), using the product of Example 12 step c).

ES 2 298 451 T3

MS (APCI) 547 (M + H⁺, 100%).

(e) Acid 7 (2R) -2-775-77 (2,3-d¿fluoro7en¿Z) -meí¿Z / í¿o7-2-meíox¿-í¿azoZo74,5-d7p¿r¿m D¿n-7-¿Z7am¿no7prop¿Z7-carbamic, 1,1-dimethyl ester

Prepared by the method of Example 1 step f), using the product of Example 12 step d).

MS (APCI) 498 (M + H⁺, 100%).

(f) 7-77 (1R) -2-Amino-1-meíiieíiZ7amino7-5-77 (2,3-difluoro7eniZ) meíiZ7íio7-iazoZo74,5-d7pyrimidin-2 (3H) -one, ir-fluoroacety

Prepared by the method of Example 1 step g), using the product of Example 12 step e) and purified by the method of Example 15 step f).

MS (APCI) 384 (M + H⁺, 100%).

NMR OH (de-DMSO) 12.55 (1H, s), 7.81 (3H, bs), 7.45-7.31 (4H, m), 7.18-7.13 (1H, m) , 4.51-4.34 (3H, m), 2.95 (2H, m), 1.14 (3H, d).

Example 13

5-77 (2,3-D¿fluoro7en¿Z) meí¿Z7í¿o7-7-77 (1R) -2-7 (2-A¿drox¿eí¿Z) am¿no7-1-meí¿Zeí ¿Z7am¿no7-í¿azoZo74,5-d7p¿r¿m¿d¿n-2 (3H) -ona, ír¿fluoroaceíaío

To a solution of the product from Example 12, step f) (100 mg) in dry THF (5 ml) was added [[(1,1-dimethylethyl) dimethylsilyl] oxy] -acetaldehyde (49 mg) followed by sodium triacetoxyborohydride ( 61 mg) and the mixture was stirred for one hour. The mixture was then acidified with concentrated hydrochloric acid, stirred at room temperature for one hour and then evaporated to dryness. The product was purified (HPLC, Novapak® C18 column, 0.1% aqueous TFA: acetonitrile, gradient elution 75:25 to 5:95 over 15 minutes) to provide the title compound (0.021 g).

MS (APCI) 428 (M + H⁺, 100%).

NMR OH (de-DMSO) 7.39-7.29 (2H, m), 7.17-12 (1H, m), 6.92 (1H, m), 4.91 (1H, s), 4 , 48-4.32 (3H, m), 3.54 (2H, m), 2.94-2.82 (4H, m), 1.12 (3H, m).

Example 14

5-77 (2,3-DZfluoro7en¿Z) meí¿Z7í¿or7-7-77 (1R) -2- (d¿meíZam¿no) -1-meí¿Zeí¿Z7am¿no7í¿azoZo74.5 -d7p¿r¿m¿d¿n-2 (3H) -ona

Prepared by the method of Example 13 using the product of Example 12 step f) and 40% aqueous formaldehyde solution.

MS (APCI) 412 (M + H⁺, 100%).

NMR OH (d₆-DMSO) 12.00 (1H, s), 7.39-7.31 (2H, m), 7.18-7.09 (2H, m), 4.39 (2H, q), 4.30 (1H, m), 3.31 (6H, bs), 2.43-2.38 (1H, m), 2.24-20 (1H, m), 1.07 (3H, d).

Example 15

5-7774- (2-Am¿noeíox¿) -3-cZoro / 'en¿Z7meí¿Z7í¿o7-7-77 (1R) -2-A¿drox¿-1-meí¿Zeí¿Z7am¿no7í¿ azoZo74,5-d7p¿r¿m¿d¿n-2 (3H) one, ír¿fluoroaceíaío (a) 2- (2-C / oro-4-form¿Z / enox¿) aceíam¿da

To a solution of 3-chloro-4-hydroxybenzaldehyde (10 g) in methanol (100 ml) was added 1.0 M potassium i-butoxide (64 ml). 2-Chloroacetamide (5.96 g) was added to the mixture and the mixture was refluxed overnight. The mixture was evaporated, the residue was triturated with water (500 ml) and the solid was collected to give the subtitle compound (4.4 g).

NMR OH (CDCl₃) 9.89 (1H, s), 7.97 (1H, d), 7.82 (1H, dd), 7.04 (1H, d), 6.73 (1H, s), 5.87 ( 1H, s), 4.63 (2H, s).

ES 2 298 451 T3 (b) 2- [2-Chloro-4- (hydroxymethyl) phenoxyJacetamide

To a solution of the product from Example 15 step a) (4.4 g) in ethanol (500 ml) was added sodium borohydride (1.56 g) and the mixture was left stirring for one hour. It was acidified with glacial acetic acid, evaporated to dryness and extracted into ethyl acetate, after which it was washed with water to give the subtitle compound (4.3 g).

NMR OH (CDCl₃) 7.44 (1H, d), 7.29 (1H, d), 6.90 (1H, d), 6.81 (1H, s), 5.85 (1H, s), 4.63 ( 2H, s), 4.48 (2H, s), 1.96 (1H, s).

(c) 2- [4 - [(Acetylthio) methylJ-2-chlorophenoxyJacetamide

Diisopropyl azocarboxylate (5.5 ml) was added to a stirred solution of triphenylphosphine (7.31 g) in THF at 0 ° C. After the addition was complete, a colorless precipitate deposited. A mixture of the product from Example 15 step b) (3.0 g) and thiol-acetic acid (2.00 ml) in THF (30 ml) at 0 ° C was added to this suspension. The mixture was allowed to reach room temperature overnight, evaporated to dryness and the residue was purified (SiO₂, 10% ethyl acetate / 90% ether as eluent) to give the subtitle compound (3.5 g)

NMR OH (CDCl₃) 7.35 (1H, d), 7.17 (1H, dd), 6.84 (1H, d), 6.76 (1H, s), 5.81 (1H, s), 4.54 ( 2H, s), 4.04 (2H, s), 2.35 (3H, s).

(d) 2- [2-Chloro-4- (mercaptomethyl) phenoxyJacetamide

Sodium hydroxide pellets (0.15 g) were added to a solution of the product from Example 15 step c) (1.0 g) in methanol (50 ml) and the mixture was stirred for 2 days. The mixture was diluted with water and the subtitle compound was collected by filtration (0.7 g).

NMR OH (d6 DMSO) 7.44 (1H, s), 7.38 (1H, d), 7.21 (1H, dd), 6.98 (1H, d), 4.55 (2H, s) , 3.76 (2H, s).

(e) 7 - [[(1R) -2-Hydroxy-1-methylethylJaminoJ-5 - [(phenyl-methyl) sulfonylJthiazolo [4,5-dJpyrimidin-2 (3H) -one

To a solution of the product of Example 3 step d) (240 mg) in acetonitrile (100 ml) and water (100 ml) was added oxone (2.4 g) and the mixture was heated at 40 ° for 2 hours. The acetonitrile was removed by rotary evaporation and the subtitle compound was collected by filtration (235 mg).

MS (APCI) 381 (M + H⁺, 100%).

(f) 5 - [[[4- (2-Aminoethoxy) -3-chlorophenylJmethylJthioJ-7 - [[(1R) -2-hydroxy-1-methylethylJaminoJthiazolo [4,5-dJpyrimidin-2 (3H) -one, trifluoroacetate

A mixture of the product of Example 15 step e) (100 mg), the product of Example 15 step d) (329 mg) and sodium borohydride (50 mg) in a solution of DMSO (1 ml) and ethanol (10 ml) heated at 55-60 ° C for 12 hours. The reaction mixture was evaporated to dryness and the residue was purified (HPLC, Novapak® C18 column, 0.1% aqueous TFA: acetonitrile, gradient elution 95: 5 to 5:95 over 15 minutes) to provide the compound of the titer (0.023 g).

MS (APCI) 442 (M + H⁺, 100%).

NMR OH (D₂O) 7.46 (1H, bs), 7.32 (1H, d), 7.00 (1H, d), 4.36-4.20 (5H, m), 3.61 (2H, m) , 3.46 (2H, m), 1.20 (3H, d).

Example 16

5 - [[(3-Chloro-4-methoxyphenyl) methylJthioJ-7 - [[(1R) -2-hydroxy-1-methylethylJaminoJthiazolo [4,5-dJpyrimidin-2 (3H) -one (a) 3-Chloro- 4-methoxybenzenemethanethiol

Thiourea (3.04 g, 0.04 mol) was added to a solution of 3-chloro-4-methoxybenzyl bromide (4.0 g), 0.02 mol) in ethanol (200 ml) and heated under reflux for 16 hours. The reaction mixture was concentrated in vacuo and the residue was subsequently dissolved in aqueous sodium hydroxide solution (30 g, 0.75 mol in 300 ml of water) and heated at 80 ° C for one hour. The reaction mixture was cooled in an ice bath and acidified by the addition of concentrated hydrochloric acid. The product was isolated by extraction 3 times in diethyl ether. The combined organic phases were dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo to give the subtitle compound as a colorless oil in 83% yield (3.0 g)

ES 2 298 451 T3

NMR dH (CDCl₃) 7.34 (1H, m,), 7.18 (1H, dd,), 6.86 (1H, d), 3.89 (3H, s), 3.68 (2H, d), 1, 76 (1H, t).

(b) 5 - [[(3-Chloro-4-methoxyphenyl] methyl) thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino] thiazolo [4,5-d] pyrimidin-2 (3H) -one

3-Chloro-4-methoxybenzenemethanethiol (0.128 g, 0.68 mmol) prepared in Example 16 step a), the product of Example 15 step e) (0.130 g, 0.349 mmol), and sodium borohydride (0.026 g, 0 , 68 mmol) were heated to reflux at 50 ° C in a mixture of dimethylsulfoxide (6 ml) and ethanol (10 ml). After 3 hours and again after 5 hours of reaction time, additional portions of sodium borohydride (0.05 g, 1.3 mmol) in ethanol (2 ml) were added to the reaction mixture and the reaction was continued Reflux at 50 ° C until conversion was complete by HPLC MS (15 hours total). The reaction mixture was neutralized by the addition of concentrated hydrochloric acid and the ethanol was removed in vacuo. The residue was purified by reverse phase chromatography on Symmetry C8, eluting with a gradient of 25% to 95% acetonitrile in 0.1M aqueous ammonium acetate over 10 minutes. The product was lyophilized from methanol / water / acetonitrile to obtain the subtitle compound in 33% yield as a white lyophilizate (0.046 g).

MS (APCI) 413 (M + H⁺, 100%).

NMR dH (de-DMSO) 12.39 (1H, bs), 7.47 (1H, m), 7.36 (1H, m), 7.25 (1H, d), 7.06 (1H, d ), 4.72 (1H, t), 4.324.21 (3H, m), 3.82 (3H, s), 3.49-3.30 (2H, m), 1.11 (3H, d) .

Example 17

5 - [[(3-Chloro-2-fluorophenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino] thiazolo [4,5-d] pyrimidin-2 (3H) -ona

a) 3-Chloro-2-fluorobenzenemethanethiol

The subtitle compound was prepared as a colorless oil in 65% yield (2.51 g) by the method described in Example 16 step a) from 3-chloro-2-fluorobenzyl bromide (5.0 g, 0.022 mol).

NMR dH (CDCl₃) 7.32-7.21 (2H, m), 7.04 (1H, t), 3.75 (2H, d), 1.90 (1H, t).

b) 5 - [[(3-Chloro-2-fluorophenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino] thiazolo [4,5-d] pyrimidin-2 ( 3H) -one

The title compound was prepared by the method of Example 16 step b) from 3-chloro-2-fluorobenzenemethanethiol, prepared in Example 17 step a), and the product of Example 15 step e).

The product was obtained in 12% yield as a white lyophilisate (0.038 g).

MP 234-5 ° C.

MS (APCI) 401 (M + H⁺, 100%).

NMR dH (de-DMSO) 12.4 (1H, bs), 7.55 (1H, m), 7.48 (1H, t), 7.26 (1H, d), 7.17 (1H, t ), 4.72 (1H, bs), 4.38 (2H, m), 4.19 (1H, m), 3.3 (2H, m), 1.08 (3H, d).

Example 18

5 - [[(2,3-Difluorophenyl) methyl] thio] -7 - [[(3R, 4R) -4-hydroxypyrrolidin-3-yl] amino] -thiazolo [4,5-d] pyrimidin-2 (3H ) -one (a) Acid 3 - [[2-amino-5 - [[(2,3-difluorophenyl) methyl] thio] -thiazolo [4,5-dJpyrimidin-7-yl] amino] -4-hydroxy- (3R, 4R) -1-pyrrolidinecarboxylic, 1,1-dimethylethyl ester

(3R, 4R) -3-amino-4-hydroxy-1-pyrrolidinecarboxylic acid, 1,1-dimethylethyl ester (0.73 g), diisopropylethyl-amine (1.0 ml) and the product of Example 4 step were stirred b), in NMP (10 ml) at 100 ° C for 28 hours. The cooled mixture was poured into water and the solid produced was collected, washed with water, and air dried. The crude material was purified (SiO2, ethyl acetate as eluent) to give the subtitle compound as a colorless solid (0.58 g), mp 182-5 ° C.

MS (APCI) 511 (M + H, 100%).

ES 2 298 451 T3 (b) Acid 3 - [[2-bromo-5 - [[(2,3-difluorophenyl) methyl] thio] -thiazolo [4,5-dJpyrimidin-7-yl] amino] -4- hydroxy- (3R, 4R) -1-pyrrolidinecarboxylic, 1,1-dimethylethyl ester

Prepared by the method of Example 1 step e), using the product of Example 18 step a).

MS (APCI) 572 (MH⁺, 100%).

(c) 3 - [[5 - [[(2,3-Difluorophenyl) methyl] thio] -2-methoxythiazolo [4,5-dJ-pyrimidin-7-yl] amino] -4-hydroxy- (3R, 4R) acid -1-pyrrolidinecarboxylic, 1,1-dimethylethyl ester

Prepared by the method of Example 1 step f), using the product of Example 18 step b).

MS (APCI) 526 (M + H⁺, 100%).

(d) 5 - [[(2,3-Difluorophenyl) methyl] thio] -7 - [[(3R, 4R) -4-hydroxypyrrolidin-3-yl] amino] thiazolo [4,5-dJpyrimidin-2 (3H ) ona

Prepared by the method of Example 1 step g), using the product of Example 18 step c). Mp 270 ° C (decomposition).

MS (APCI) 412 (M + H⁺, 100%).

NMR δΗ (d6-DMSO) 7.32 (2H, m), 7.14 (1H, m), 6.46 (1H, d), 5.57 (1H, s), 4.39 (2H, s ), 4.30 (2H, m), 3.39 (2H, m), 3.12 (1H, dd), 2.98 (1H, d),

Example 19

5 - [[(2,3-Difluorophenyl) methyl] thio] -7 - [(3R) -pyrrolidin-3-ylamino] thiazolo [4,5-dJpyrimidin-2 (3H) -one, dihydrochloride (a) Acid 3 - [[2-amino-5 - [[(2,3-difluorophenyl) methyl] thio] -thiazolo [4,5-dJpyrimidin-7-yl] amino] - (3R) -1-pyrrolidinecarboxylic ester 1,1 -dimethylethyl

Prepared by the method of Example 18 step a), using (R) -3-amino-1-pyrrolidinecarboxylic acid, 1,1-dimethylethyl ester and the product of Example 4 step b).

MS (APCI) 495 (M + H⁺, 100%).

(b) 3 - [[2-Bromo-5 - [[(2,3-difluorophenyl) methyl] thio] -thiazolo [4,5-dJ-pyrimidin-7-yl] amino] - (3R) -1-pyrrolidinecarboxylic acid , 1,1-dimethylethyl ester

Prepared by the method of Example 1 step e), using the product of Example 19 step a).

MS (APCI) 559 (M + H⁺, 100%).

(c) 3 - [[5 - [[(2,3-Difluorophenyl) methyl] thio] -2-methoxythiazolo [4,5-dJ-pyrimidin-7-yl] amino] - (3R) -1-pyrrolidinecarboxylic acid, ester 1,1-dimethylethyl

Prepared by the method of Example 1 step f), using the product of Example 19 step b).

MS (APCI) 510 (M + H⁺, 100%).

(d) 5 - [[(2,3-Difluorophenyl) methyl] thio] -7 - [(3R) -pyrrolidin-3-ylamino] thiazolo [4,5-dJpyrimidin-2 (HR) -one, dihydrochloride

Prepared by the method of Example 1 step g), using the product of Example 19 step c) which was then converted to the salt.

MP 178-181 ° C

MS (APCI) 396 (M + H⁺, 100%).

ES 2 298 451 T3

NMR δΗ (de-DMSO) 12.75 (1H, s), 9.19 (2H, bd), 7.91 (1H, d), 7.37 (2H, m), 7.17 (1H, m ), 4.66 (1H, m), 4.43 (2H, dd), 3.10-3.50 (4H, m), 2.17 (1H, m), 1.96 (1H, m) .

Example 20

7 - [[(1R) -2-Hydroxy-1-methylethyl] amino-5 - [[(2-methyl-4-thiazolyl) methyl] thio] thiazolo [4,5-dJpyrimidin-2 (3H) -one ( a) 6-Amino-2 - (((2-methyl-4-thiazolyl) methyl) thio) -4 (3H) -pyrimidinone

Hydrated 4-amino-6-hydroxy-2-mercaptopyrimidine (16.1 g) and powdered sodium hydroxide (8.0 g) were stirred in dry DMF (100 ml) for 20 min. 4-Chloromethyl-2-methylthiazole hydrochloride monohydrate (20 g) was added little by little and the resulting suspension was stirred for 18 hours. The mixture was poured into water and the solid was collected, washed with water and dried to provide the subtitle compound (24.3 g).

MS (APCI) 255 (M + H⁺, 100%).

(b) 2-Amino-5 - [[(2-methyl-4-thiazolyl) methyl] thio] -thiazolo- [4,5-dJpyrimidin-7 (6H) -one

The product from Example 20 step a) (24.3 g) and potassium thiocyanate (37.1 g) were stirred in dry DMF (400 ml) with pyridine (13.1 ml) at 0 ° C. Bromine (4.5 ml) was added over one hour. After stirring for two hours, the mixture was poured into water. The resulting solution was concentrated to a low volume and then water was added. The resulting solid was collected, adsorbed on 2M hydrochloric acid, and precipitated by adding saturated sodium bicarbonate solution. The solid was collected, washed with water and dried to give the subtitle compound (8.7 g).

MS (APCI) 312 (M + H⁺, 100%).

(c) 7-Chloro-5 - [[(2-methyl-4-thiazolyl) methyl] thio] thiazolo- [4,5-dJpyrimidin-2-amine

Prepared by the method of Example 1 step c), using the product of Example 20 step b) (4.3 g).

MS (APCI) 330/332 (M + H⁺), 330 100%).

(d) (2R) -2 - [[(2-Amino-5 - [[(2-methyl-4-thiazolyl) methyl] thio] -thiazolo [4,5-dJ-pyrimidin-7-yl) amino] -1 -propanol

Prepared by the method of Example 18 step a), using the product of Example 20 step c).

MP 220-2 ° C

MS (APCI) 369 (M + H⁺, 100%).

(e) (2R) -2 - [[2-Bromo-5 - [[(2-methyl-4-thiazolyl) methyl] thio] -thiazolo [4,5-dJ-pyrimidin-7-yl] amino] -1- propanol

Prepared by the method of Example 1 step e), using the product of Example 20 step d).

MS (APCI) 443 (M + H⁺, 100%).

(f) (2R) -2 - [[2-Methoxy-5 - [[(2-methyl-4-thiazolyl) methyl] thio] -thiazolo [4,5-dJpyrimidin-7-yl] amino] -1- propanol

Prepared by the method of Example 1 step f), using the product of Example 20 step e).

MS (APCI) 384 (M + H⁺, 100%).

(g) 7 - [[(1R) -2-Hydroxy-1-methylethyl] amino] -5 - [[(2-methyl-4-thiazolyl) methyl] thio] thiazolo [4,5-d] pyrimidin-2 (3H) -one

Prepared by the method of Example 1 step g), using the product of Example 20 step f).

MP 208-9 ° C

MS (APCI) 370 (M + H⁺, 100%).

ES 2 298 451 T3

NMR OH (d6-DMSO) 12.37 (1H, s), 7.35 (1H, s), 7.32 (1H, d), 4.73 (1H, t), 4.36 (2H, s ), 4.21 (1H, m), 3.38 (2H, m), 2.62 (3H, s), 1.10 (3H, d).

Example 21

7 - [[2-Hydroxy-1- (hydroxymethyl) ethyl] amino] -5 - [[(2-methyl-4-thiazolyl) methyl] thio] -thiazolo [4,5-d] -pyrimidm-2 (3H ) -one (a) 2 - [[(2-Amino-5 - [[(2-methyl-4-thiazolyl) methyl] thio] -thiazolo [4,5-d] pyrimidin-7-yl) amino] - 1,3-propanediol

Prepared by the method of Example 18 step a), using the product of Example 20 step c) and 2-amino-1,3-propanediol.

mp 158-160 ° C

MS (APCI) 385 (M + H, 100%).

(b) 2 - [[2-Bromo-5 - [[(2-methyl-4-thiazolyl) methyl] thio] -thiazolo [4,5-d] pyrimidin-7-yl] amino] -1,3- propanediol

Prepared by the method of Example 1 step e), using the product of Example 21 step a).

MS (APCI) 448 (M + H⁺, 100%).

(c) 2 - [[2-Methoxy-5 - [[(2-methyl-4-thiazolyl) methyl] thio] -thiazolo [4,5-d] pyrimidin-7-yl] amino] -1,3- propanediol

Prepared by the method of Example 1 step f), using the product of Example 21 step b).

MS (APCI) 400 (M + H⁺, 100%).

(d) 7 - [[2-Hydroxy-1- (hydroxymethyl) ethyl] ammo] -5 - [[(2-methyl-4-thiazolyl) methyl] thio] thiazolo [4,5-d] pyrimidin-2 ( 3H) -one

Prepared by the method of Example 1 step g), using the product of Example 21 step c). mp 239-243 ° C

MS (APCI) 386 (M + H⁺, 100%).

NMR OH (d6-DMSO) 12.37 (1H, s), 7.38 (1H, s), 7.24 (1H, d), 4.67 (2H, t), 4.36 (2H, s ), 4.20 (1H, m), 3.50 (4H, m), 2.62 (3H, s).

Example 22

7 - [(2-Hydroxy-1,1-dimethylethyl) amino] -5 - [[(2-methyl-4-thiazolyl) methyl] thio] -thiazolo [4,5-d] pyrimidin-2 (3H) - one (a) 2 - [[2-Amino-5 - [[(2-methyl-4-thiazolyl) methyl] thio] -thiazolo [4,5-d] pyrimidin-7-yl) amino] -2-methyl -1-propanol

Prepared by the method of Example 18 step a), using the product of Example 20 step c) and 2-amino-2-methylpropanol

mp 250-252 ° C

MS (APCI) 383 (M + H, 100%).

(b) 2 - ([2-Bromo-5 - ([(2-methyl-4-thiazolyl) methyl] thio) -thiazolo [4,5-d] pyrimidin-7-yl] amino] -2-methyl- 1-propanol

Prepared by the method of Example 1 step e), using the product of Example 22 step a).

MS (APCI) 446 (M + H⁺, 100%).

(c) 2 - [[2-Methoxy-5 - [[(2-methyl-4-thiazolyl) methyl] thio] -thiazolo [4,5-d] pyrimidin-7-yl] amino] -2-methyl- 1-propanol

Prepared by the method of Example 1 step f), using the product of Example 22 step b).

MS (APCI) 398 (M + H⁺, 100%).

ES 2 298 451 T3 (d) 7 - [(2-Hydroxy-1,1-dimethylethyl) amino] -5 - [[(2-methyl-4-thiazolyl) methyl] thio] -thiazolo [4,5-dJpyrimidine -2 (3H) -one

Prepared by the method of Example 1 step g), using the product of Example 22 step c).

MP 231-2 ° C

MS (APCI) 384 (M + H⁺, 100%).

NMR δΗ (d₆-DMSO) 12.36 (1H, s), 7.37 (1H, s), 6.61 (1H, bs), 4.80 (1H, t), 4.37 (2H, s), 3, 55 (2H, d), 2.62 (3H, s), 1.31 (6H, s).

Example 23

7 - [(2-Hydroxy-1,1-dimethylethyl) ammo] -5 - [[(2-methylpheml) -methyl] thio] -thiazolo [4,5-dJpyrimidm-2 (3H) -one (a) 7 - [(2-Hydroxy-1,1-dimethylethyl) amino] -5 - [(phenyl-methyl) sulfonyl] thiazolo [4,5-dJpyrimidin-2 (3H) -one

A stirred solution of the product from Example 1 step g) (0.14 g) in glacial acetic acid (30 ml) was treated with peracetic acid (30-40% in acetic acid, 2 ml), stirred for 2 h, and then at 50 ° C for 1 h. The solution was quenched with an excess of dimethyl sulfide and evaporated to give a gum.

MS (APCI) 395 (M + H⁺, 100%).

(b) 7 - [(2-Hydroxy-1,1-dimethylethyl) ammo] -5 - [[(2-methyl-pheml) methyl] thio] -thiazolo [4,5-dJpyrimidm-2 (3H) -one

The product from Example 23 step a) was taken up in DMSO (1.73 ml), treated with ¹ potassium butoxide and divided into 3 portions. A portion was treated with 2-methylphenylmethyl-mercaptan (0.053 g), stirred at 50 ° C for 1 hr and 2 hr, neutralized with glacial acetic acid, and subjected to reverse phase preparative HPLC on a C8 Symmetry 19 column. x 50 mm using 10 to 60% acetonitrile in 0.1% aqueous ammonium acetate for 6 min at 20 ml / min to give the title compound.

MS (APCI) 377 (M + H⁺, 100%).

NMR δH (d6-DMSO) 1.33 (s, 6H); 2.35 (s, 3H); 3.57 (d, 2H); 4.33 (s, 2H); 4.82 (t, 1H); 6.57 (broad s, 1H); 7.127.20 (mult., 3H); 7.41 (d, 1H); 12.37 (broad s, 1H).

Example 24

5 - [(2-Furanylmethyl) thiol-7-1 [(1R) -2-hydroxy-1-methylethyl] -amino] -thiazolo [4,5-dJpyrimidin-2 (3H) -one (a) 7- [ [(1R) -2-Hydroxy-1-methylethyl] amino] -5-1- (phenyl-methyl) sulfonyl] -thiazolo [4,5-dJpyrimidin-2 (3H) -one

The subtitle compound was prepared from the product of Example 3 step d), using the method of Example 23 step a).

MS (ES) 381 (M + H⁺, 100%).

(b) 5 - [[(2-Furanylmethyl) thio] -7 - [[(1R) -2-hydroxy-1-methyl-ethyl] amino] -thiazolo [4,5-dJpyrimidin-2 (3H) -one

The title compound was prepared from the product of Example 24 step (a), using the method of Example 23 step (b) using furfuryl mercaptan.

MS (APCI) 339 (M + H⁺, 100%).

NMR δH (d4-methanol) 1.12 (d, 3H); 3.41-3.45 (mult., 1H); 3.49-3.53 (mult., 1H); 4.24-4.32 (mult, 3H); 6.18-6.22 (mult., 2H); 7.29 (broad s, 1H),

Example 25

7 - [[(1R) -2-Amino-1-methylethyl] amino] -5 - [[(3-chloro-2-fluorophenyl) methyl] thio] -thiazolo [4,5-dJpyrimidin-2 (3H) - one (a) [(1R) -2-amino-1-methyl-2-oxoethyl] carbamic acid, 9H-fluoren-9-ylmethyl ester

A solution of D-alaninamide hydroxychloride (3 g) in 10% sodium carbonate solution (50 ml) and dioxane (50 ml) was treated with FMOC chloride (6.24 g) in dioxane (40 ml) and stirred overnight. Mix

ES 2 298 451 T3 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) BP311 (+ H) (b) [(1R) -2-amino-1-methylethylJcarbamic acid, 9H-fluoren-9-ylmethyl ester

To a solution of the product from Example 25 step a) (6.9 g) in THF (100 ml) was added borane-methyl sulfide complex (4.4 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) BP 297 (+ H) (c) (2R) -12- (9H) -fluoren-9-ylmethoxycarbonylamino) -propyl) carbamic acid, 1,1-dimethylethyl ester

To a stirred solution of the product from Example 25 step b) (3.0 g) in THF (100 ml) was added ditert-butyl dicarbonate (2.2 g) 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).

OH NMR (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-AminopropylJcarbamic acid, ester 1,1-dimethylethyl

Piperidine (5 ml) was added to a solution of the product from Example 25 step c) (3.8 g) in THF (100 ml) and the mixture was allowed to stand for one 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 (1.7 g).

OH NMR (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-amino-5 - [(phenylmethyl) thioJthiazolo- [4,5-dJpyrimidin-7-ylJaminoJpropylJcarbamic acid, 1,1-dimethylethyl ester

The product from Example 1 step c) (2.0 g) and the product from Example 25 step d) (1.3 g) in an NMP solvent (10 ml) containing Hunig's base (3 ml) was heated to 110 ° C for 10 hours. The mixture was evaporated to dryness and purified (SiO₂, dichloromethane: ethyl acetate (1: 1) as eluent) to give the subtitle compound (1.9 g).

MS (ESI) BP 447 (+ H).

(f) [(2R) -2 - [[2-amino-5 - [(phenylmethyl) sulfonylJthiazolo [4,5-dJpyrimidin-7-ylJaminoJpropylJcarbamic acid, 1,1-dimethylethyl ester

To a solution of OXONE (7.0 g) in water (400 ml) was added sodium hydrogen carbonate until the pH was adjusted to 7.4. A solution of the product from Example 25 step e) (1.9 g) in acetonitrile (100 ml) was added to this solution and the mixture was heated at 4 ° C for 2 hours. After completion of the reaction, the acetonitrile was removed by rotary evaporation to give the subtitle compound (1.7 g).

MS (ESI) BP 479 (+ H).

(g) 3-Chloro-2-fluoro-benzenemethanethiol

A mixture of 3-chloro-2-fluorobenzyl bromide (5.0 g), thiourea (3.4 g) in ethanol solvent (200 ml) was refluxed for 16 hours. The mixture was evaporated to dryness and a solution of sodium hydroxide (30 g) in water (300 ml) was added to the residue and the mixture was heated under reflux for one hour. Allow to cool to room temperature and acidify with concentrated hydrochloric acid, extract the product into ether to give the subtitle compound as an oil (2.51 g).

ES 2 298 451 T3

NMR dH (CDCl₃) 7.32-21 (2H, m), 7.04 (1H, t), 3.75 (2H, d), 1.90 (1H, t).

(h) [(2R) -2 - [[2-amino-5 - [[(3-chloro-2-fluorophenyl) -methyl] thio] thiazolo [4,5-d] pyrimidin-7-yl] amino acid ] propyl] carbamic, 1,1-dimethylethyl ester

To a mixture of the product of Example 25 step f) (1.2 g), the product of Example 25 step g) (1.6 g) in a mixed solvent of ethanol (30 ml) and DMSO (5 ml) was added sodium borohydride (100 mg) and the mixture was heated at 50 ° C for 2 hours. The ethanol was removed by rotary evaporation and the crude product was extracted into ethyl acetate and washed with water. The subtitle compound was obtained by purification (SiO₂, (1: 1) dichloromethane: ethyl acetate as eluent) to give (1.95 g).

MS (ESI) PB 499 (+ H).

(i) [(2R) -2 - [[2-bromo-5 - [[(3-chloro-2-fluorophenyl) -methyl] thio] thiazolo [4,5-dJpyrimidin-7-yl] amino] propyl acid ] carbamic, 1,1-dimethylethyl ester

Prepared by the method of Example 1 step e), using the product of Example 25 step h).

MS (APCI) 562 (M + H⁺, 100%).

(j) [(2R) -2 - [[5 - [[(3-chloro-2-fluorophenyl) methyl] -thio] -2-methoxythiazolo [4,5-dJpyrimidin-7-yl] amino] propyl] acid -carbamic, 1,1-dimethylethyl ester

Prepared by the method of Example 1 step f), using the product of Example 25 step i).

MS (APCI) 514 (M + H⁺, 100%).

(k) 7 - [[(1R) -2-Amino-1-methylethyl] amino] -5 - [[(3-chloro-2-fluorophenyl) methyl] thio] thiazolo [4,5-dJpyrimidin-2 (3H ) -ona

Prepared by the method of Example I step g), using the product of Example 25 step j).

MP 241-3 ° C

MS (APCI) 400 (M + H⁺, 100%).

NMR dH (de-DMSO) 7.56 (1H, m), 7.49 (1H, m), 7.17 (1H, m), 7.05 (1H, bs), 4.44 (1H, m ), 4.39 (2H, ab), 2.92 (2H, d), 1.13 (3H, d).

Example 26 (2S) -2 - [[5 - [[(2,3-Difluorophenyl) methyl] thio] -2,3-dihydro-2-oxothiazolo [4,5-dJpyrimidin-7-yl] amino] -3 -hydroxy-propanamide (a) (2S) -2 - [[2-Amino-5 - [[(2,3-difluorophenyl) methyl] thio] -thiazolo [4,5-dJ-pyrimidin-7-yl] amino] - 3-hydroxy-propanamide

The subtitle compound was prepared according to Example 2 step a) using the product from Example 4 step b) (2 g, 6 mmol), 1-serinamide (0.66 g, 6 mmol), NMP (80 ml) , and diisopropylethylamine (2 ml) to give the subtitle compound (1.36 g).

MP 145-151 ° C

MS (APCI) 413 (M + H⁺, 100%).

NMR dH (de-DMSO) 8.10 (2H, bis), 7.40-7.07 (6H, m), 4.57 (1H, q), 4.43 (1H, d), 4.36 (1H, d), 3.71 (2H, d).

(b) (2S) -2 - [[5 - [[(2,3-Difluorophenyl) methyl] thio] -2,3-dihydro-2-oxothiazolo [4,5-dJpyrimidin-7-yl] amino] - 3-hydroxypropanamide

Prepared by consecutive use of the methods of Example 1 Steps e), f) and g), using the product of Example 26 step (a). Compounds formed during the separate steps were not purified or characterized.

ES 2 298 451 T3

MS (APCI) 414 (M + H⁺, 100%).

NMR OH (d₆-DMSO) 12.47 (1H, br), 7.47 (1H, br), 7.42 (1H, s), 7.34 (2H, m), 7.13 (1H, m), 7, 09 (1H, s), 4.90 (1H, t), 4.58 (1H, m), 4.39 (2H, m), 3.70 (2H, m).

Example 27

7-77 (¿R) -2-H¿drox¿-1-meí¿ / eí¿ / 7 am¿no7-5-7 (2-í¿en¿ / meí¿ /) - í¿o7í¿azo / o74 , 5-d7p¿r¿m¿d¿n-2 (3H) -one (a) 7-77 (1R) -2-H¿drox¿-1-meí¿ / eí¿ / 7 am¿no7-5- 7 (2-í¿en¿ / meí¿ /) í¿o7í¿azo / o74,5-d7p¿r¿m¿d¿n-2 (3H) -ona

The title compound was prepared by the method described in Example 16 step b) from the product of Example 15 step e) (0.300 g, 0.79 mmol) and 2-thiophenomethanethiol (0.32 ml, 3.9 mmol ).

The product was obtained in a low yield of 3% as a white lyophilisate (0.010 g).

MS (APCI) 355 (M + H⁺, 100%).

NMR OH (d₆-DMSO) 12.50 (1H, bs), 7.36 (1H, m), 7.16 (1H, bs), 7.07 (1H, m), 6.92 (1H, m), 4, 72 (1H, bs), 4.55 (2H, d), 4.26 (1H, m), 3.44 (2H, m), 1.12 (3H, d).

Example 28

7-77 (1R) -2-H¿drox¿-1-meí¿ / eí¿ / 7 am¿no7-5-7773-meí¿ / -4- (meí¿ / sufon¿ /) / en¿ / 7meí¿ / 7í¿o7í¿azo / o74,5-d7p¿r¿m¿d¿n-2 (3H) ona (a) 3-Meí¿ / -4- (meí¿ / í¿o) benza / dehyde

Tin (IV) chloride (13.6 ml, 0.116 mol) was added to an ice bath cooled solution of 1-methyl-2 (methylthio) benzene (10 g, 0.073 mol) in anhydrous dichloromethane (200 ml) under nitrogen and stirred for a further 2 hours at 0 ° C. Α, α-Dichloromethylmethyl ether (6.56 ml, 0.073 mol) was introduced and the reaction mixture was stirred for one hour at &lt;10 ° C before removing the cooling. After reaching room temperature, the reaction mixture was poured into ice / water (400 ml), stirred and then extracted with dichloromethane (x 3). The combined organic layers were dried over anhydrous magnesium sulfate, filtered, concentrated over silica gel, and purified by flash chromatography, eluting with diethyl ether / isohexane (10: 1) to give the subtitle compound as a brown oil (6 , 54 g) in 54% yield.

GCMS 166 (M⁺, 100%).

NMR Oh (CDCl₃) 9.91 (1H, s), 7.68 (1H, m), 7.62 (1H, s), 7.24 (1H, t), 2.54 (3H, s), 2.36 ( 3H, s).

(b) 3-Meí¿ / -4- (meí¿ / í¿o) benzenomeíanoZ

Sodium borohydride (1.40 g, 0.037 mol) was added to an ice bath cooled solution of the product from Example 28 step a) (6.16 g, 0.037 mol) in ethanol (50 ml). After one hour, the reaction mixture was neutralized by the careful addition of aqueous hydrochloric acid (2 molar) and concentrated in vacuo to remove the organic solvent. The remaining aqueous solution was then extracted with ethyl acetate (x 3). The combined organic layers were dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the subtitle compound as a brown oil (6 g) in quantitative yield.

GCMS 168 (M⁺, 100%).

NMR oh (CDCl₃) 7.18 (3H, m), 4.62 (2H, bs), 2.46 (3H, s), 2.33 (3H, s).

(c) 3-Meí¿ / -4- (meí¿ / su / fon¿ /) benzenemean /

3-Chloroperoxybenzoic acid (57-86% grade, 20.4 g) was stirred in dichloromethane (150 ml), dried over anhydrous magnesium sulfate and then filtered. The filtrate was added dropwise over one hour to a stirred, ice-bath-cooled solution of the product from Example 28 step b) (5.67 g, 0.034 mol) in dichloromethane (50 ml). The reaction mixture was filtered and the filtrate was washed with aqueous sodium hydrogen carbonate solution followed by aqueous sodium dithionite solution (10 g Na₂OR₄S₂ in 150 ml of water). The organic layer was dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo before purification by flash chromatography, elution with dichloromethane / methanol (100: 2). The subtitle compound was obtained as a yellow oil (5.52 g) in 82% yield.

ES 2 298 451 T3

MS (APCI) 201.1 (M + H⁺, 94.3%).

NMR δ_Η (CDCl₃) 7.87 (1H, d), 7.38 (2H, m), 5.40 (1H, q), 4.56 (2H, d), 3.18 (3H, s), 2.61 ( 3H, s).

(d) 3-Methyl-4- (methylsulfonyl) benzenemethanethioacetate

Diethyl azodicarboxylate (4.33 ml, 0.028 mol) was added to an ice bath cooled solution of triphenylphosphine (7.20 g, 0.028 mol) in tetrahydrofuran (40 ml). A solution of the product of Example 28 step c) (5.5 g, 0.028 mol) dissolved in tetrahydrofuran (20 ml) was added to the resulting suspension. After the precipitate had dissolved, thiolacetic acid was added to the reaction solution and cooling was suppressed. After 16 hours at room temperature, the reaction mixture was concentrated on silica gel and purified by flash chromatography, eluting with isohexane / ethyl acetate (2: 1). The subtitle compound was obtained as a pink solid (2.46 g) in 35% yield. NMR δΗ (de-DMSO) 7.84 (1H, d), 7.36 (2H, m), 4.16 (2H, s), 3.19 (3H, s), 2.61 (3H, s) ), 2.37 (3H, s).

(e) bis [[3-Methyl-4- (methylsulfonyl) phenyl] methyl] -disulfide

A mixture of the product from Example 28 step d) (1.98 g, 7.66 mmol) and 7 molar methanolic ammonia (30 ml) was stirred for 24 hours. The product precipitated out of solution as a white solid and was isolated by filtration, after which it was dried in vacuo. The filtrate was treated analogously with 7 molar ammonia in methanol and provided a second crop of white solid product. In total, the subtitle compound was obtained in 32% yield (0.534 g).

MS (APCI) 451 (M + NH₄⁺, 98.9%).

NMR δΗ (d6-DMSO) 7.88 (2H, s), 7.38-7.34 (4H, m), 3.88 (4H, s), 3.20 (6H, s), 2.64 (6H, s).

(f) 7 - [[(1R) -2-hydroxy-1-methylethyl] amino] -5 - [[[3-methyl-4- (methylsulfonyl) phenyl] methyl] thio] thiazolo [4,5-d] pyrimidin-2 (3H) -one

The title compound was prepared by the method described in Example 16 step b) using the product from Example 15 step e) (0.20 g, 0.53 mmol) and the product from Example 28 step e) (0.34 g, 0.79 mmol) to give 11% product as a white lyophilisate (0.025 g).

MS (APCI) 441 (M + H⁺, 100%).

NMR δΗ (d6-DMSO) 12.40 (1H, s), 7.81 (1H, d), 7.52 (2H, m), 7.33 (1H, d), 4.74 (1H, t ), 4.35 (2H, s), 4.19 (1H, m), 3.41 (1H, m), 3.34-3.28 (1H, m), 3.18 (3H, s) , 2.61 (3H, s), 1.08 (3H, d).

Example 29

5 - [[[3-Chloro-4- (trifluoromethoxy) phenyl] methyl] thio] -7 - [[(1R) -2-hydroxy -1-methylethyl] amino] thiazolo [4,5- ^ 0-0 ^ ^ 2- (3H) -one (a) 3-Chloro-4- (trifluoromethoxy) benzenemethanethiol

To a solution of 3-chloro-4- (trifluoromethoxy) benzyl bromide (5 g) in ethanol (100 ml) was added thiourea (5 g) and the mixture was heated under reflux for 2 hours. The mixture was evaporated to dryness and the residue was taken up in water (100 ml). Sodium hydroxide pellets (3 g) were added to this solution and the mixture was heated under reflux for one hour. The mixture was allowed to cool to room temperature and was acidified with concentrated hydrochloric acid, the mixture was extracted with ether, dried and evaporated to give the subtitle compound as a colorless waxy solid (3.5 g).

NMR δΗ (CDCl3) 7.35-7.09 (3Η, m), 3.58 (2Η, s).

(b) 5 - [[[3-Chloro-4- (trifluoromethoxy) phenyl] methyl] thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino] thiazolo [4,5-d] pyrimidin2 (3H) -one

The title compound was prepared by the method described in Example 16 step b) using the product from Example 15 step e) (0.40 g, 1.05 mmol) and the product from Example 29 step a) (0.71 g, 1.5 mmol) to give 10% product as a white lyophilisate (0.046 g).

MS (APCI) 467 (M + H⁺, 100%).

ES 2 298 451 T3

NMR Oh (d6-DMSO) 12.42 (1H, s), 7.75 (1H, m), 7.52 (2H, m), 7.43 (1H, d), 4.72 (1H, t ), 4.34 (2H, d), 4.18 (1H, quint), 3.46-3.27 (2H, m), 1.07 (3H, d).

Example 30

5 - [[[2-Fluoro-3- (trifluoromethyl) feml] methyl] thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino] thiazolo [4,5-dJpyrimidin-2 (3H ) one (a) 2-Fluoro-3- (trifluoromethyl) benzenemethanethiol

The subtitle compound was prepared from 2-fluoro- (3-trifluoromethyl) benzyl bromide (10 g) using the method of Example 29 step a).

NMR Oh (CDCl₃) 7.68-7.18 (3H, m), 3.74 (2H, s), 1.98 (1H, s).

(b) 5 - [[[2-Fluoro-3- (trifluoromethyl) phenyl] methyl] thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino] thiazolo [4,5-d] pyrimidin2 (3H) -one

The title compound was prepared by the method described in Example 16 step b) using the product from Example 15 step e) (0.47 g, 1.23 mmol) and the product from Example 30 step a) (0.775 g, 3.7 mmol) to give 5% product as a white lyophilisate (0.025 g).

MS (APCI) 435 (M + H⁺, 100%).

NMR oh (d6-DMSO) 12.42 (1H, s), 7.92 (1H, t), 7.68 (1H, t), 7.35 (2H, m), 4.71 (1H, bs ), 4.42 (2H, m), 4.16 (1H, quintet), 3.40-3.30 (2H, m), 1.07 (3H, d).

Example 31

5 - [[(2,3-Difluorophenyl) methyl] thio] -7- [2 - [(dimethylamino) -ethyl] amino] thiazolo [4,5-d] pyrimidin-2 (3H) -one, monohydrochloride (a ) 2-Bromo-7-chloro-5 - [[(2,3-difluorophenyl) methyl] thio] -thiazolo [4,5-d] pyrimidine

The product from Example 4 step b) (8.0 g) was suspended in bromoform (200 ml) followed by addition of tert-butyl nitrite (8 ml) and the whole was heated at 60 ° C for 30 minutes. The solvents were removed under reduced pressure and the residue was purified by column chromatography (silica-dichloromethane / isohexane 1: 1) to give a yellow solid (5.6 g).

MS (APCI) 409/411 (M + H, 100%).

(b) 7-Chloro-5 - [(2,3-difluorophenyl) methylthio] -2-methoxythiazolo [4,5-d] pyrimidine

The product from Example 31 step a) (5.6 g) was suspended in methanol (150 ml) and potassium hydroxide powder (0.77 g) was added. The whole was stirred at room temperature for 2 hours. The mixture was adjusted to pH 7 with a few drops of concentrated hydrochloric acid before being evaporated to dryness. It was purified by column chromatography (silica-isohexane / dichloromethane 3: 2 to 1: 1) to give a white solid (2.0 g).

MS (APCI) 360/362 (M + H, 100%).

(c) 7-Chloro-5 - [[(2,3-difluorophenyl) methyl] thio] thiazolo [4,5-d] pyrimidin-2 (3H) -one

The product from Example 31 step b) (2.0 g) was dissolved in dioxane (150 ml) followed by the addition of concentrated hydrochloric acid (1 ml) and water (1 ml), and the whole was heated at 40 ° C for 67 hours. The mixture was evaporated to dryness and purified by column chromatography (silica-dichloromethane) to give a white solid (1.4 g).

MS (APCI) 346/348 (M + H, 100%).

ES 2 298 451 T3 (d) 5 - [[(2,3-Difluorophenyl) methyl] thio] -7- [2 - [(dimethylamino) ethyl] amino] -thiazolo [4,5-dJpyrimidin-2 (3H) -one, monohydrochloride

The product from Example 31 step (c) (1.4 g) was dissolved in dry tetrahydrofuran (5 ml) and N, N-dimethylethylenediamine (0.25 g) was added to the solution in a finger pump which was heated to 80 ° C for 24 hours. Solvents were removed under reduced pressure and the residue was partitioned between ethyl acetate and brine. The combined organic extracts were dried (sodium sulfate) and evaporated under reduced pressure so that the subsequent residue was purified by column chromatography (silica-ethyl acetate / methanol 5: 1) to give the free base as a sticky solid ( 0.095 g). This was converted to the monohydrochloride by suspending the solid in methanol (10 ml) followed by adding concentrated hydrochloric acid (3 drops) to ensure dissolution, followed by water (100 ml) to lyophilize the compound to obtain a brown powder. (0.080 g).

Mp 263 ° C (decomposition)

MS (APCI) 398 (M + H, 100%).

NMR δΗ (d₆-DMSO) 12.57 (1H, s), 10.22 (1H, s), 7.94 (1H, t), 7.40 (1H, m), 7.34 (1H, m), 7, 16 (1H, m), 4.43 (2H, s), 3.78 (2H, s), 3.21 (2H, m), 2.78 (6H, d)

Example 32

5 - [[(2-Fluorophenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methyl-ethyl] amino] -thiazolo [4,5-dJpyrimidin-2 (3H) -one ( a) 2 - [[2-Amino-5 - [(phenylmethyl) thio] thiazolo [4,5-dJpyrimidin-7-yl] amino] - (2R) -1-propanol

The product from Example 1 step c) (25.0 g), D-alaninol (12.3 g) and diisopropylethylamine (26.0 g) were diluted in N-methylpyrrolidinone (250 ml) and stirred at 100 ° C for 24 hours before cooling and pouring the reaction mixture into water (2.5 L). The precipitate was filtered and dried in vacuo before being pre-absorbed on silica gel. Chromatography using EtOAc, 4% MeOH / EtOAc as eluent provided the desired product as a yellow solid (9.0 g, 32%).

MS (APCI) 347 (M + H, 100%).

(b) 2 - [[2-Amino-5-mercaptothiazolo [4,5-dJpyrimidin-7-yl] -amino] - (2R) -1-propanol

Sodium metallic was added little by little to a solution of the product from Example 32 step a) (5.0 g) in ammonia (150 ml) until a blue tint persisted. Then ammonium chloride was added and the solvent was allowed to evaporate. The residue was dissolved in water (200 ml) and filtered before neutralization with 2M HCl solution. The gray precipitate was filtered, washed with water (200 ml) and dried in vacuo for 48 h to give the subtitle compound as a brown solid (3.0 g).

MS (APCI) 258 (M + H, 100%).

(c) 2 - [[2-Amino-5 - [[(2-fluorophenyl) methyl] thio] thiazolo [4,5-dJpyrimidin-7-yl] amino] - (2R) -1-propanol

2-Fluorobenzyl bromide (0.369 g) was added little by little to a solution of the product from Example 32 step b) (0.5 g) and diisopropylethylamine (0.26 g) in DMSO / N-methylpyrrolidinone (4 ml / 0 , 5 ml) at 50 ° C and stirring was maintained for one hour. The mixture was partitioned between water (200 ml) and EtOAc (120 ml). The organic phase was recovered and further washed with water (200 ml), dried over MgSO₄ and concentrated on silica gel. The subtitle compound was purified by flash chromatography using DCM and then EtOAc as eluents to give a white solid (245 mg, 35%).

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

(d) 2 - [[2-Bromo-5 - [[(2-fluorophenyl) methyl] thio] thiazolo [4,5-dJpyrimidin-7-yl] amino] - (2R) -1-propanol

Isoamyl nitrite (0.3 ml) was added to a suspension of the product from Example 32 step c) (0.23 g) in bromoform (15 ml) and acetonitrile (15 ml) at 50 ° C. Stirring was maintained for 10 min before concentrating to approximately 3 ml. The residue was purified by column chromatography using 20% ​​EtOAc / DCM as eluent to give the subtitle compound as a yellow solid (102mg, 38%).

MS (APCI) 429 (M + H, 100%).

ES 2 298 451 T3 (e) 2 - [[5 - [[(2-Fluorophenyl) methylJthioJ-2-methoxythiazolo- [4,5-dJpyrimidin-7-ylJaminoJ- (2R) -1-propanol

Potassium hydroxide (27 mg) was added to a solution of the product from Example 32 step d) (0.1 g) in MeOH (10 ml). The mixture was stirred for 24 hr before neutralizing to pH 7 with 2M HCl solution. Volatiles were removed in vacuo and the product was used directly in the next step.

MS (APCI) 381 (M + H, 100%).

(f) 5 - [[(2-Fluorophenyl) methylJthioJ-7 - [[(1R) -2-hydroxy-1-methylethylJaminoJ-thiazolo [4,5-dJpyrimidin-2 (3H) -one

The product from Example 32 step e) was dissolved in 1,4-dioxane (50 ml), water (1 ml) and concentrated HCl solution (0.5 ml) and stirred for 20 h at 40 ° C. Volatiles were removed under reduced pressure and the crude product was purified by preparative HPLC to provide the subtitle compound as a white solid (21 mg).

MS (APCI) 367 (M + H, 100%)

NMR OH (d₆-DMSO) 12.40 (1H, s), 8.14-7.11 (5H, m), 4.72 (1H, t), 4.35 (2H, m), 4.22 (1H, m ), 3.47-3.29 (2H, m), 1.10 (3H, d)

Example 33

7 - [[(1R) -2-Hydroxy-1-methylethylJaminoJ-5 - [[(2-methoxyphenyl) methylJthioJ-thiazolo [4,5-dJpyrimidin-2 (3H) -one (a) 2 - [[2- Amino-5 - [[(2-methoxyphenyl) methylJthioJthiazolo [4,5-dJpyrimidin-7-ylJaminoJ- (2R) -1-propanol

Prepared by the method of Example 32 step c), using the product of Example 32 step b).

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

(b) 2 - [[2-Bromo-5 - [[(2-methoxyphenyl) methylJthioJthiazolo [4,5-dJpyrimidin-7-ylJaminoJ- (2R) -1-propanol

Prepared by the method of Example 32 step d), using the product of Example 33 step a).

MS (APCI) 441 (M + H⁺, 100%).

(c) 2 - [[2-Methoxy-5 - [[(2-methoxyphenyl) methylJthioJthiazolo- [4,5-dJpyrimidin-7-ylJaminoJ- (2R) -1-propanol

Prepared by the method of Example 32 step e), using the product of Example 33 step b).

MS (APCI) 393 (M + H⁺, 100%).

(d) 7 - [[(1R) -2-Hydroxy-1-methylethylJaminoJ-5 - [[(2-methoxy-phenyl) methylJthioJ-thiazolo [4,5-dJpyrimidin-2 (3H) -one

Prepared by the method of Example 32 step f), using the product of Example 33 step c).

MS (APCI) 379 (M + H⁺, 100%).

NMR OH (d6-DMSO) 7.40 (1H, dd), 7.22 (1H, dt), 6.97 (1H, d), 6.84 (1H, dt), 6.00 (1H, d ), 4.25 (2H, m), 4.15 1H, m), 3.83 (3H, s), 3: 48-3.31 (2H, m), 1.10 (3H, d).

Example 34

7 - [[(1R) -2-Hydroxy-1-methylethylJaminoJ-5 - [(2-phenoxyethyl) -thioJthiazolo [4,5-dJpyrimidin-2 (3H) -one (a) 2 - [[2-Amino- 5 - [(2-phenoxyethyl) thioJthiazolo [4,5-dJpyrimidin-7-ylJaminoJ- (2R) -1-propanol

Prepared by the method of Example 32 step c), using the product of Example 32 step b).

MS (APCI) 378 (M + H⁺, 100%).

ES 2 298 451 T3 (b) 2 - [[2-Bromo-5 - [(2-phenoxyethyl) thio] thiazolo [4,5-dJpyrimidin-7-yl] amino] - (2R) -1-propanol

Prepared by the method of Example 32 step d), using the product of Example 34 step a).

MS (APCI) 441 (M + H⁺, 100%).

(c) 2 - [[2-Methoxy-5 - [(2-phenoxyethyl) thio] thiazolo [4,5-dJpyrimidin-7-yl] amino] - (2R) -1-propanol

Prepared by the method of Example 32 step e), using the product of Example 34 step b).

MS (APCI) 393 (M + H⁺, 100%).

(d) 7 - [[(1R) -2-Hydroxy-1-methylethyl] amino] -5 - [(2-phenoxyethyl) thio] thiazolo [4,5-dJpyrimidin-2 (3H) -one

Prepared by the method of Example 32 step f), using the product of Example 34 step c).

MS (APCI) 379 (M + H⁺, 100%).

NMR δH (d6-DMSO) 12.37 (1H, s), 7.30-7.26 (3H, m), 6.96-6.91 (3H, m), 4.71 (1H, t) , 4.23-4.14 (3H, m), 3.463.28 (4H, m), 1.08 (3H, d)

Example 35

7 - [[(1R) -2-Hydroxy-1-methylethyl] amino] -5 - [[(3-methylphenyl) -methyl] thio] -thiazolo [4,5-dJpyrimidin-2 (3H) -one (a ) 2 - [[2-Amino-5 - [[(3-methylphenyl) methyl] thio] thiazolo [4,5-dJpyrimidin-7-yl] amino] - (2R) -1-propanol

Prepared by the method of Example 32 step c), using the product of Example 32 step b).

MS (APCI) 362 (M + H⁺, 100%).

(b) 2 - [[2-Bromo-5 - [[(3-methylphenyl) methyl] thio] thiazolo [4,5-dJpyrimidin-7-yl] amino] - (2R) -1-propanol

Prepared by the method of Example 32 step d), using the product of Example 35 step a).

MS (APCI) 425 (M + H⁺, 100%).

(c) 2 - [[2-Methoxy-5 - [[(3-methylphenyl) methyl] thio] thiazolo [4,5-dJpyrimidin-7-yl] amino] - (2R) -1-propanol

Prepared by the method of Example 32 step e), using the product of Example 35 step b).

MS (APCI) 377 (M + H⁺, 100%).

(d) 7 - [[(1R) -2-Hydroxy-1-methylethyl] amino] -5 - [[(3-methyl-phenyl) methyl] thio] -thiazolo [4,5-dJpyrimidin-2 (3H) -ona

Prepared by the method of Example 32 step f), using the product of Example 35 step c).

MS (APCI) 363 (M + H⁺, 100%).

NMR δH (d₆-DMSO) 12.37 (1H, s), 7.23-7.16 (4H, m), 7.04 (1H, d), 4.73 (1H, t), 4.28 (2H, m ), 4.24 (1H, m),

3.48-3.30 (2H, m), 2.28 (3H, s), 1.11 (3H, d).

Example 36

5 - [[(2-Fluoro-3-methylphenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino] -thiazolo [4,5-dJpyrimidin-2 (3H) - one (a) 2 - [[2-Amino-5 - [[(2-fluoro-3-methylphenyl) methyl] thio] thiazolo [4,5-dJpyrimidin-7-yl] amino] - (2R) -1- propanol

Prepared by the method of Example 32 step c), using the product of Example 32 step b).

MS (APCI) 380 (M + H⁺, 100%).

ES 2 298 451 T3 (b) 2 - [[2-Bromo-5 - [[(2-fluoro-3-methylphenyl) methyl] thio] -thiazolo [4,5-dJpyrimidin-7-yl] amino] - ( 2R) -1-propanol

Prepared by the method of Example 32 step d), using the product of Example 36 step a).

MS (APCI) 443 (M + H⁺, 100%).

(c) 2 - [[5 - [[(2-Fluoro-3-methylphenyl) methyl] thio] -2-methoxythiazolo [4,5-dJpyrimidin-7-yl] amino] - (2R) -1-propanol

Prepared by the method of Example 32 step e), using the product of Example 36 step b).

MS (APCI) 395 (M + H⁺, 100%).

(d) 5 - [[(2-Fluoro-3-methylphenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino] thiazolo [4,5-dJpyrimidin-2 (3H ) -ona

Prepared by the method of Example 32 step f), using the product of Example 36 step c).

MS (APCI) 381 (M + H⁺, 100%).

NMR dH (de-DMSO) 12.39 (1H, s), 7.37-7.00 (4H, m), 4.72 (1H, t), 4.33 (2H, m), 4.22 (1H, m), 3.47-3.30 (2H, m), 2.23 (3H, s), 1.11 (3H, d)

Example 37

5 - [[(3-Chlorophenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methyl-ethyl] amino] -thiazolo [4,5-dJpyrimidin-2 (3H) -one ( a) 2 - [[2-Amino-5 - [[(3-chlorophenyl) methyl] thio] thiazolo [4,5-dJpyrimidin-7-yl] amino] - (2R) -1-propanol

Prepared by the method of Example 32 step c), using the product of Example 32 step b).

MS (APCI) 382 (M + H⁺, 100%).

(b) 2 - [[2-Bromo-5 - [[(3-chlorophenyl) methyl] thio] thiazolo [4,5-dJpyrimidin-7-yl] amino] - (2R) -1-propanol

Prepared by the method of Example 32 step d), using the product of Example 37 step a).

MS (APCI) 445 (M + H⁺, 100%).

(c) 2 - [[5 - [[(3-chlorophenyl) methyl] thio] -2-methoxythiazolo [4,5-dJpyrimidin-7-yl] amino] - (2R) -1-propanol

Prepared by the method of Example 32 step e), using the product of Example 37 step b).

MS (APCI) 397 (M + H⁺, 100%).

(d) 5 - [[(3-chlorophenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino-thiazolo [4,5-dJpyrimidin-2 (3H) -one

Prepared by the method of Example 32 step f), using the product of Example 37 step c).

MS (APCI) 383 (M + H⁺, 100%).

NMR dH (d₆-DMSO) 12.40 (1H, s), 7.49 (1H, d), 7.43-7.30 (4H, m), 4.72 (1H, t), 4.32 (2H, m ), 4.21 (1H, m),

3.48-3.26 (2H, m), 1.09 (3H, d).

Example 38

5 - [[(3-Bromophenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methyl-ethyl] amino] -thiazolo [4,5-dJpyrimidin-2 (3H) -one ( a) 2 - [[2-Amino-5 - [[(3-bromophenyl) methyl] thio] thiazolo [4,5-dJpyrimidin-7-yl] amino] - (2R) -1-propanol

Prepared by the method of Example 32 step c), using the product of Example 32 step b).

MS (APCI) 426 (M + H⁺, 100%).

ES 2 298 451 T3 (b) 2-772-Bromo-5-77 (3-bromophen¿ /) meí¿ / 7í¿or7í¿azo / o74,5-d7p¿r¿m¿d¿n-7-¿ / 7 am? No7- (2R) -1-propane /

Prepared by the method of Example 32 step d), using the product of Example 38 step a).

MS (APCI) 491 (M + H⁺, 100%).

(c) 2-775-77 (3-Bromofen¿ /) meí¿ / 7í¿o7-2-meíox¿í¿azo / o74,5-d7p¿r¿m¿d¿n-7-¿/ 7 am¿ no7- (2R) -1-propane /

Prepared by the method of Example 32 step e), using the product of Example 38 step b).

MS (APCI) 443 (M + H⁺, 100%).

(d) 5-77 (3-Bromofen¿ /) meí¿ / 7í¿o7-7-77 (¿R) -2-A¿drox¿-1-meí¿ / eí¿ / 7 am¿no7-í¿azo / o74,5-d7p¿r¿m¿d¿n-2 (3H) -one

Prepared by the method of Example 32 step f), using the product of Example 38 step c).

MS (APCI) 427 (M + H⁺, 100%).

NMR OH (d₆-DMSO) 12.40 (1H, s), 7.63 (1H, t), 7.46-7.24 (4H, m), 4.72 (1H, t), 4.31 (2H, m ), 4.21 (1H, m),

3.48-3.26 (2H, m), 1.10 (3H, d)

Example 39

5-7774-CDfluoromeíox¿fen¿ / 7meí¿ / 7í¿o7-7-77 (1R) -2-h¿drox¿-1-meí¿ / eí¿ / 7 am¿no7í¿azo / o74,5-d7p¿ r¿m¿d¿n-2 (3H) -one (a) 2-772-Am¿no-5-7774- (d¿fluoromeíox¿) phen¿ / 7meí¿ / 7í¿o7-í¿azo / o74 , 5-d7p¿r¿m¿d¿n-7-¿/ 7 am¿no7- (2R) -1-propane /

Prepared by the method of Example 32 step c), using the product of Example 32 step b).

MS (APCI) 414 (M + H⁺, 100%).

(b) 2-772-Bromo-5-7774- (d¿fluoromeíox¿) phen¿ / 7meí¿ / 7í¿o7-í¿azo / o74,5-d7p¿r¿m¿d¿n-7-¿ / 7 am? No7- (2R) -1-propane /

Prepared by the method of Example 32 step d), using the product of Example 39 step a).

MS (APCI) 477 (M + H⁺, 100%).

(c) 2-775-7774-CD¿fluoromeíox¿lfen¿ / 7meí¿ / 7í¿o7-2-meíox¿í¿azo / o74,5-d7p¿r¿m¿d¿n-7-¿/ 7am No7- (2R) -1-propane /

Prepared by the method of Example 32 step e), using the product of Example 39 step b).

MS (APCI) 429 (M + H⁺, 100%).

(d) 5-7774- (D¿fluoromeíox¿) phen¿ / 7meí¿ / 7í¿o7-7-77 (1R) -2-A¿drox¿-1-meí¿ / eí¿ / 7 am¿no7í¿azo / o74,5-d7p¿r¿m¿d¿n-2 (3H) one

Prepared by the method of Example 32 step f), using the product of Example 39 step c).

MS (APCI) 415 (M + H⁺, 100%).

NMR OH (d6-DMSO) 12.38 (1H, s), 7.48 (2H, dt), 7.26 (1H, d), 7.19 (1H, t), 7.11 (2H, dd ), 4.73 (1H, t), 4.31 (2H, m), 4.21 (1H, m), 3.47-3.30 (2H, m), 1.10 (3H, d)

Example 40 (+/-) - 5-77 (2,3-D¿fluorofen¿ /) meí¿ / 7í¿o7-7-772-A¿drox¿-1- (meíox¿meí¿ /) eí¿ / 7 am¿no7í¿azo/o74,5-d7p¿r¿m¿d¿n-2 (3H) -one (a) (+/-) - 2-Am¿no-3-meíox¿-1-propane / , A¿droc / oruro

To a suspension of DL-3-methoxy-alanine (1.0 g) in dry THF (100 ml) was added borane-methyl sulfide complex (10 ml) and the mixture was heated under reflux for 16 hours. The mixture was then quenched with methanol while

ES 2 298 451 T3 was refluxed, evaporated to dryness and the residue was taken up in methanolic hydrogen chloride (100 ml) and refluxed for a further 2 hours, after which it was evaporated to dryness to give the compound of the subtitle as a colorless gum (1.0 g).

NMR Ó_H (D₂O) 3.40 (3H, s), 3.53-3.74 (4H, m), 3.81 (1H, m).

(b) (+/-) - 2 - [[2-amino-5 - [[(2,3-difluorophenyl) methyl] -thio] ti-azolo [4,5-d] pyrimidin-7-yl] amino ] -3-methoxy-1-propanol

Prepared by the method of Example 12 step a), using the product of Example 4 step b) and the product of Example 40 step a).

MS (APCI) 414 (M + H⁺, 100%).

(c) (+/-) - 2 - [[2-Chloro-5 - [[(2,3-difluorophenyl) methyl] thio] -thiazolo [4,5-d] pyrimidin-7-yl] amino] - 3-methoxy-1-propanol

To a solution of the product of Example 40 step b) (1.0 g) in a mixture of concentrated hydrochloric acid (40 ml) and water (32 ml) cooled in an ice bath was added a solution of sodium nitrite (0 , 4 g) in water (5 ml), and stirred at this temperature for 2 hours. The mixture was then extracted into ethyl acetate, dried and evaporated to give the subtitle compound (0.6 g).

MS (APCI) 434 (M + H⁺, 100%).

(d) (+/-) - 2 - [[5 - [[(2,3-Difluorophenyl) methyl] thio] -2-methoxythiazolo [4,5-d] pyrimidin-7-yl] amino] -3- methoxy-1-propanol

Prepared by the method of Example 1 step f), using the product of Example 40 step c).

MS (APCI) 429 (M + H⁺, 100%).

(e) (+/-) - 5 - [[(2,3-Difluorophenyl) methyl] thio] -7 - [[2-hydroxy-1- (methoxymethyl) ethyl] amino] thiazolo [4,5-d] pyrimidin-2 (3H) one

Prepared by the method of Example 1 step g), using the product of Example 40 step d).

MS (APCI) 415 (M + H⁺, 100%).

Example 41

7 - [[2-Hydroxy-4- (hydroxymethyl) ethyl] amino] -5 - [(phenylmethyl) thio] thiazolo [4,5-d] pyrimidin-2 (3H) -one (a) 2 - [[2 -Amino-5 - [(phenylmethyl) thio] thiazolo [4,5-d] pyrimidin-7-yl] amino] -1,3-propanediol

Prepared by the method of Example 12 step a), using the product of Example 1 step c) and 2-amino-1,3-propanediol.

MS (APCI) 364 (M + H⁺, 100%).

NMR OH (d₆-DMSO) 7.42-7.38 (1H, m), 7.28 (1H, t), 7.22 (1H, t), 5.30 (1H, d), 4.63 (1H, bs ), 4.28 (2H, s), 4.03 (1H, m), 3.54-3.40 (4H, m).

(b) 2 - [[2-Chloro-5 - [(phenylmethyl) thio] thiazolo [4,5-d] pyrimidin-7-yl] amino] -1,3-propanediol

Prepared by the method of Example 40 step c), and the product of Example 41 step a).

MS (APCI) 384 (M + H⁺, 100%).

(c) 2 - [[2-Methoxy-5 - [(phenylmethyl) thio] thiazolo [4,5-d] pyrimidin-7-yl] amino] -1,3-propanediol

Prepared by the method of Example 1 step f), and the product of Example 41 step b).

MS (APCI) 379 (M + H⁺, 100%).

ES 2 298 451 T3 (d) 7 - [[2-hydroxy-1- (hydroxymethyl) ethyl] amino] -5 - [(phenylmethyl) thio] -thiazolo [4,5-d] pyrimidin-2 (3H) - ona

Prepared by the method of Example 1 step g), and the product of Example 41 step c).

MS (APCI) 365 (M + H⁺, 100%).

Example 42 5 - [[(2-Bromophenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methyl-ethyl] amino] -thiazolo [4,5-dJpyrimidin-2 (3H) - one (a) 2 - [[2-Amino-5 - [[(2-bromphenyl) methyl] thio] thiazolo [4,5-dJpyrimidin-7-yl] amino] - (2R) -1-propanol

Prepared by the method of Example 32 step c), using the product of Example 32 step b).

MS (APCI) 428 (M + H⁺, 100%).

(b) 2 - [[2-Bromo-5 - [[(2-bromophenyl) methyl] thio] thiazolo [4,5-dJpyrimidin-7-yl] amino] - (2R) -1-propanol

Prepared by the method of Example 1 step e), using the product of Example 42 step a).

MS (APCI) 491 (M + H⁺, 100%).

(c) 2 - [[5 - [[(2-Bromophenyl) methyl] thio] -2-methoxythiazolo [4,5-dJpyrimidin-7-yl] amino] - (2R) -1-propanol

Prepared by the method of Example 1 step f), using the product of Example 42 step b).

MS (APCI) 443 (M + H⁺, 100%).

(d) 5 - [[(2-Bromophenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino] -thiazolo [4,5-d] pyrimidin-2 (3H) -ona

Prepared by the method of Example 1 step g), using the product of Example 42 step c).

MS (APCI) 427 (M + H⁺, 100%).

NMR δΗ (d₆-DMSO) 12.41 (1H, s), 7.65-7.14 (5H, m), 4.72 (1H, t), 4.42 (2H, s), 4.21 (1H, m ), 3.47-3.30 (2H, m), 1.10 (3Η, ά)

Example 43

5 - [[(2,3-Difluorophenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino] -thiazolo [4,5-dJpyrimidin-2 (3H) -one, sodium salt

The product from Example 5 step d) was suspended in water and 1 equivalent of 0.1 N sodium hydroxide solution was added to this suspension, followed by the addition of a small aliquot of tetrahydrofuran to promote dissolution. The resulting solution was then lyophilized to give the title compound as a colorless solid.

MP 218-220 ° C

MS (APCI) 385 (M + ff, 100%).

NMR δΗ (d6-DMSO) 7.39-7.09 (3Η, m), 5.60 (1Η, d), 4.65 (1Η, m), 4.34 (2Η, q), 4.09 (1Η, m), 3.44-3.27 (2Η, m), 1.06 (3Η, d).

Example 44

5 - [[(3-Chloro-2-bromophenyl) methyl] thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino] -thiazolo [4,5-dJpyrimidin-2 (3H) - one, sodium salt

Prepared as in Example 43 using the product from Example 17 step b).

MS (APCI) 401 (M + II ', 100%).

ES 2 298 451 T3

Example 45 (+/-) - 5 - [[(2,3-Difluorophenyl) methyl] thio] -7 - [[2-hydroxy-1- (methoxymethyl) ethyl] amino] -thiazolo [4,5-d] pyrimidin-2 (3H) -one, sodium salt

Prepared by the method of Example 43 using the product of Example 40 step e).

MP&gt; 250 ° C

MS (APCI) 415 (M + H⁺, 100%).

NMR δH (de-DMSO) 7.39-7.04 (3H, m), 5.51 (1H, d), 4.68 (1H, t), 4.34 (2H, q), 4.22 (1H, m), 3.51-3.35 (4H, m), 3.32 (3H, s).

Example 46

7 - [[(2-Hydroxy-1- (hydroxymethyl) ethyl) amino] -5 - [(phenylmethyl) thio] thiazolo [4,5-d] pyrimidin-2 (3H) -one, sodium salt

Prepared by the method of Example 43 using the product of Example 41 step d).

MP 231-2 ° C

MS (APCI) 365 (M + H⁺, 100%).

NMR δH (de-DMSO) 7.41-7.18 (5H, m), 5.30 (1H, d), 4.63 (2H, s), 4.28 (2H, s), 4.06 (1H, m), 3.50 (4H, m).

Example 47

7 - [[(1R) -2-Hydroxy-1-methylethyl] -amino] -5 - [(phenylmethyl) -thio] thiazolo [4,5-dJpyrimidin-2 (3H) -one, sodium salt

Prepared by the method of Example 43 using the product of Example 3 step d).

MP (shrinks at 110) melts at 221-225 ° C

MS (APCI) 349 (M + H⁺ 100%).

NMR δH (de-DMSO) 7.41-7.18 (5H, m), 5.58 (1H, d), 4.65 (1H, t), 4.28 (2H, q), 4.11 (1H, m), 3.49-3.31 (2H, m), 1.08 (3H, d).

Example 48

5 - [(5-Chloro-1,2,3-thiadiazol-4-yl) thio] -7 - [[(1R) -2-hydroxy-1-methylethyl] amino] thiazolo [4,5-dJpyrimidin-2 (3H) -one (a) (2R) -2 - [[2-Amino-5 - [(5-chloro-1,2,3-thiadiazol-4-yl) -thio] thiazolo [4,5-dJpyrimidin -7-yl] amino] -1-propanol

Prepared by the method of Example 32 step c), using the product of Example 32 step b) and 5-chloro-4 (chloromethyl) -1,2,3-thiadiazole.

MS (APCI) 390 (M + H⁺, 100%).

(b) (2R) -2 - [[2-Chloro-5 - [[(5-chloro-1,2,3-thiadiazol-4-yl) -methyl] thio] thiazolo [4,5-dJpyrimidin-7 -yl] amino] -1-propanol

Prepared by the method of Example 40 step c) and using the product of Example 48 step a).

MS (APCI) 409 (M + H⁺, 100%).

(c) (2R) -2 - [[(5 - [[(5-Chloro-1,2,3-thiadiazol-4-yl) methyl] -thio] -2-methoxythiazolo [4,5-dJpyrimidin-7 -yl] amino] -1-propanol

Prepared by the method of Example 1 step f) and using the product of Example 48 step b).

MS (APCI) 405 (M + H⁺, 100%).

ES 2 298 451 T3 (d) 5 - [[(5-Chloro-1,2,3-thiadiazol-4-yl) thiol-7 - [[(1R) -2-hydroxy-1-methylethylJaminoJ-thiazolo [4 , 5-dJpyrimidin-2 (3H) one

Prepared by the method of Example 1 step g) and using the product of Example 48 step c).

MS (APCI) 391 (M + H⁺, 100%).

NMR OH (d6-DMSO) 12.39 (1H, s), 7.39 (1H, d), 4.76 (2H, AB), 4.70 (1H, t), 4.24 (1H, m ), 3.48-3.30 (2H, m), 1.11 (3H, d).

Pharmacological data

Ligand Binding Assay

It was purchased [¹²⁵I] IL-8 (human, recombinant) from Amersham, UK, with a specific activity of 2000 Ci / mmol. All other chemicals were analytical grade. High levels of hrCXCR2 were expressed in HEK293 cells (human embryonic kidney 293 cells, ECACC No. 85120602) (Lee et al. (1992) J. Biol. Chem. 267 pp 16293-16291). 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 cloned using Qiagen Megaprep 2500 and transfected into HEK 293 cells using Lipofectamine reagent (Gibco BRL). Cells of the maximally expressing 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 phenyl-methyl sulfonyl fluoride, 2 pg / ml soybean trypsin inhibitor, 3 mM benzamidine, 0.5 pg / ml leupeptin and 100 pg / ml bacitracin)] and the cells were allowed to swell for 10 minutes. The cell preparation was disrupted using a glass hand mortar homogenizer with a PTFE handle and the cell membranes were collected 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 0.45 µm 96-well MultiScreen 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, MgCl₂ 1 mM, 0.125 mg / ml of Bacitracin and 0.1% (w / v) of gelatin]. Additionally, a compound of formula (I) according to the examples was predissolved in DMSO and added until reaching a final concentration of 1% (v / v) of DMSO. The assay was started with the addition of membranes and after 1.5 hours at room temperature the membranes were harvested by filtration using a Millipore MultiScreen vacuum manifold and washed twice with assay buffer (no bacitracin). The backing plate was removed from the MultiScreen plate assembly, and the filters were dried at room temperature, punched out, and then counted in a Cobra-γ-counter.

Compounds of formula (I) according to the examples were found to have CI values₅₀ less than (&lt;) 10 pM.

Intracellular Calcium Mobilization Assay

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

GROa (human, recombinant) chemokine was purchased from R&amp;D Systems (Abingdon, UK). All other products were of analytical quality. Changes in free intracellular 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, pp. 513-519). The cells were loaded for one hour at 37 ° C in loading buffer (storage buffer with 0.1% (w / v) gelatin) containing 5 pM fluo-3 AM ester, washed with loading buffer and They were then resuspended in Tyrode's salt solution supplemented with 5.7 mM glucose, 0.1% (w / v) bovine serum albumin (BSA), CaCl₂ 1.8 mM and MgCl₂ 1 mM. Cells were pipetted into 96-well black-walled, light-bottomed 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 the addition of a concentration A₅₀ of GROa and the transient increase in fluorescence of fluo-3

ES 2 298 451 T3 (λ_Εχ = 490 nm and l_Em = 520 nm) was monitored using a FLIPR instrument (Fluorometric Imaging Plate Reader, Molecular Devices, Sunnyvale, USA).

The compounds of formula (I) according to the examples were tested and found to be antagonists of the CXCR2 receptor on human neutrophils.