WO2008017688A1 - 2-carboxy thiophene derivatives as anti-viral agents - Google Patents

Abstract

Anti-viral agents of compounds of Formula (I) wherein A, R1, R2 and R3 are as defined in the specification, processes for their preparation and their use in HCV treatment are provided.

Description

2-CARBOXY THIOPHENE DERIVATIVES AS ANTI-VIRAL AGENTS

FIELD OF THE INVENTION

The present invention relates to novel 2-carboxy thiophene derivatives useful as anti-viral agents. Specifically, the present invention involves novel inhibitors of Hepatitis C Virus (HCV) replication.

BACKGROUND OF THE INVENTION

Infection with HCV is a major cause of human liver disease throughout the world. In the US, an estimated 4.5 million Americans are chronically infected with HCV. Although only 30% of acute infections are symptomatic, greater than 85% of infected individuals develop chronic, persistent infection. Treatment costs for HCV infection have been estimated at $5.46 billion for the US in 1997. Worldwide over 200 million people are estimated to be infected chronically. HCV infection is responsible for 40-60% of all chronic liver disease and 30% of all liver transplants. Chronic HCV infection accounts for 30% of all cirrhosis, end-stage liver disease, and liver cancer in the U.S. The CDC estimates that the number of deaths due to

HCV will minimally increase to 38,000/year by the year 2010.

Due to the high degree of variability in the viral surface antigens, existence of multiple viral genotypes, and demonstrated specificity of immunity, the development of a successful vaccine in the near future is unlikely. Alpha-interferon (alone or in combination with ribavirin) has been widely used since its approval for treatment of chronic HCV infection. However, adverse side effects are commonly associated with this treatment: flu-like symptoms, leukopenia, thrombocytopenia, depression from interferon, as well as anemia induced by ribavirin (Lindsay, K. L. (1997) Hepatology 26 (suppl 1 ): 71 S-77S). This therapy remains less effective against infections caused by HCV genotype 1 (which constitutes -75% of all HCV infections in the developed markets) compared to infections caused by the other 5 major HCV genotypes. Unfortunately, only -50-80% of the patients respond to this treatment (measured by a reduction in serum HCV RNA levels and normalization of liver enzymes) and, of responders, 50-70% relapse within 6 months of cessation of treatment. Recently, with the introduction of pegylated interferon (Peg-IFN), both initial and sustained response rates have improved substantially, and combination treatment of Peg-IFN with ribavirin constitutes the gold standard for therapy. However, the side effects associated with combination therapy and the impaired response in patients with genotype 1 present opportunities for improvement in the management of this disease.

First identified by molecular cloning in 1989 (Choo, Q-L et al (1989) Science 244:359-362), HCV is now widely accepted as the most common causative agent of post-transfusion non A, non-B hepatitis (NANBH) (Kuo, G et al (1989) Science 244:362-364). Due to its genome structure and sequence homology, this virus was assigned as a new genus in the Flaviviridae family. Like the other members of the Flaviviridae, such as flaviviruses (e.g. yellow fever virus and Dengue virus types 1-4) and pestiviruses (e.g. bovine viral diarrhea virus, border disease virus, and classic swine fever virus) (Choo, Q-L et al (1989) Science 244:359-362; Miller, R.H. and R.H. Purcell (1990) Proc. Natl. Acad. Sci. USA 87:2057-2061 ), HCV is an enveloped virus containing a single strand RNA molecule of positive polarity. The HCV genome is approximately 9.6 kilobases (kb) with a long, highly conserved, noncapped 5' nontranslated region (NTR) of approximately 340 bases which functions as an internal ribosome entry site (IRES) (Wang CY et al 'An RNA pseudoknot is an essential structural element of the internal ribosome entry site located within the hepatitis C virus 5' noncoding region' RNA- A Publication of the RNA Society. 1 (5): 526-537, 1995 JuL). This element is followed by a region which encodes a single long open reading frame (ORF) encoding a polypeptide of -3000 amino acids comprising both the structural and nonstructural viral proteins.

Upon entry into the cytoplasm of the cell, this RNA is directly translated into a polypeptide of -3000 amino acids comprising both the structural and nonstructural viral proteins. This large polypeptide is subsequently processed into the individual structural and nonstructural proteins by a combination of host and vi rally-en coded proteinases (Rice, CM. (1996) in B.N. Fields, D.M.Knipe and P.M. Howley (eds) Virology 2^nd Edition, p931-960; Raven Press, N.Y.). Following the termination codon at the end of the long ORF, there is a 3' NTR which roughly consists of three regions: an - 40 base region which is poorly conserved among various genotypes, a variable length poly(U)/polypyrimidine tract, and a highly conserved 98 base element also called the "3' X-tail" (Kolykhalov, A. et al (1996) J. Virology 70:3363-3371 ; Tanaka, T. et al (1995) Biochem Biophys. Res. Commun. 215:744-749; Tanaka, T. et al (1996) J. Virology 70:3307-3312; Yamada, N. et al (1996) Virology 223:255-261 ). The 3' NTR is predicted to form a stable secondary structure which is essential for HCV growth in chimps and is believed to function in the initiation and regulation of viral RNA replication.

The NS5B protein (591 amino acids, 65 kDa) of HCV (Behrens, S. E. et al (1996) EMBO J. 15:12-22), encodes an RNA-dependent RNA polymerase (RdRp) activity and contains canonical motifs present in other RNA viral polymerases. The NS5B protein is fairly well conserved both intra-typically (-95-98% amino acid (aa) identity across 1 b isolates) and inter-typically (-85% aa identity between genotype 1 a and 1 b isolates). The essentiality of the HCV NS5B RdRp activity for the generation of infectious progeny virions has been formally proven in chimpanzees (A. A. Kolykhalov et al.. (2000) Journal of Virology, 74(4): 2046-2051 ). Thus, inhibition of NS5B RdRp activity (inhibition of RNA replication) is predicted to be useful to treat HCV infection.

Although the predominant HCV genotype worldwide is genotype 1 , this itself has two main subtypes, denoted 1a and 1 b. As seen from entries into the Los Alamos HCV database (www.hcv.lanl.gov) (Table 1 ) there are regional differences in the distribution of these subtypes: while genotype 1 a is most abundant in the United States, the majority of sequences in Europe and Japan are from genotype 1 b. Table 1

Based on the foregoing, there exists a significant need to identify synthetic or biological compounds for their ability to inhibit replication of both genotype 1 a and genotype 1 b of HCV.

PCT publication number WO2002/100851 generically discloses certain compounds, including certain 2-carboxy thiophene compounds, having HCV inhibitory activity. The data provided relates to an HCV polymerase assay utilising the 1 b genotype. The compounds disclosed have the formula (I) v-Yy^s _VR'

Z XX X (D wherein

X is chosen from -N(R³)M(R²) or -JN(R²)(R³);

M is chosen from -SO₂-, -SO-, -S-, -C(O)-, -C(S)-, -CH₂C(O)N(R⁴)-, -CH₂C(S)N(R¹⁵)-, -

CH(R¹⁵)-, -C(=N(R⁸))-, or a bond;

R⁴ is Ci_-6alkyl;

R⁸ is chosen from H, Ci_i₂alkyl, C_2-i₂alkenyl, C_2-i₂alkynyl, C₆-i₄aryl, C_3-i₂heterocycle, C_3- i₂heteroaralkyl, C_6-i₆aralkyl;

R¹⁵ is chosen from H or d-βalkyl;

J is chosen from -C(W)-, -C(R⁶)-, -S-, -S(O)-, or -SO₂-;

W is chosen from O, S or NR⁷;

R⁷ is chosen from H, Ci_i₂alkyl, C_2-i₂alkenyl, C_2-i₂alkynyl, C₆-i₄aryl, C_3-i₂heterocycle, C_3- i₂heteroaralkyl, C_6-i₆aralkyl;

R⁶ is chosen from H, Ci_i₂alkyl, C₆-i₄aryl, or C_6-i₆aralkyl;

Y¹ is chosen from a bond, Ci_-6alkyl, C_2-6alkenyl, C_2-6alkynyl;

Y is chosen from COOR¹⁶, COCOOR⁵, P(0)0ROR^b, S(O)OR⁵, S(O)₂OR⁵, tetrazole,

CON(R⁹)CH(R⁵)COOR⁵, CONR¹⁰R¹¹, CON(R⁹)-SO₂-R⁵, CONR⁹OH, or halogen;

R⁹, R⁵, R¹⁰ and R¹¹ are each independently chosen from H, Ci_i₂alkyl, C_2-i₂alkenyl, C_2-

₁₂alkynyl, C_3-i₂heterocycle, C_3-i₈heteroaralkyl, C_6-i₈aralkyl; or R¹⁰ and R¹¹ are taken together with the nitrogen to form a 3 to 10 membered heterocycle;

R^a and R^b are each independently chosen from H, Ci_i₂alkyl, C_2-i₂alkenyl, C_2-i₂alkynyl, C_6- i₄aryl, C_3-i₂heterocycle, C_3-i₈heteroaralkyl, C_6-i₈aralkyl; or R^a and R^b are taken together with the oxygens to form a 5 to 10 membered heterocycle; R¹⁶ is chosen from H, Ci.-i₂alkyl, C₂-i₂alkenyl, C₂-i₂alkynyl, C_6-i₄aryl, C_3-i₂heterocycle, C_3- iβheteroaralkyl, C_6-i₈aralkyl; provided that R¹⁶ is other than methyl or ethyl; R¹ is chosen from Ci.-i₂alkyl, C_2-i₂alkenyl, C_2-i₂alkynyl, C_6-i₄aryl, C_3-i₂heterocycle, C_3- iβheteroaralkyl, C_6-i₈aralkyl; R² is chosen from Ci.-i₂alkyl, C_2-i₂alkynyl, C₆-i₄aryl, C_3-i₂heterocycle, C_3-i₈heteroaralkyl, C_6- iβaralkyl;

R³ is chosen from H, Ci_i₂alkyl, C_2-i₂alkenyl, C_2-i₂alkynyl, C₆-i₄aryl, C_3-i₂heterocycle, C_3- iβheteroaralkyl, C_6-i₈aralkyl; and Z is chosen from H, halogen, or d-βalkyl.

PCT publication number WO2003/062215 generically discloses certain compounds, including certain 2-carboxy thiophene compounds, having kinase inhibiting activity. The compounds disclosed have the fo

wherein V is, inter alia, a group of formula (i)

R¹ represents T, N-(CO)R⁶R⁷, N(R⁶)COR⁷, or NC(O)OR⁶R⁷; T represents H, (hetero)alkyl, alkenyl, cycloalkyl, (hetero)aryl, arylalkyl or alkylaryl; R⁸ represents T, NR³R⁴, N-C(O)R³R⁴, N(R³)COR⁴, OR³, COR³, CO₂R³ or OCOR³; R², R⁵ represent T, alkylaryl, carboxyl, halo or CF₃; R³, R⁴, R⁶, R⁷ represent T, alkylaryl, COOR⁵ or COR⁵; X, Y, Z represent CH or N; U represents S or NH; and W represents NH, O or S.

PCT publication number WO2004/052885 generically discloses certain compounds, including certain 2-carboxy thiophene compounds, having HCV inhibitory activity. The data provided relates to an HCV polymerase assay utilising the 1 b genotype. The compounds disclosed have the formula (I)

wherein Z is chosen from 3-7 membered heterocycle or 3-7 membered cycloalkyl; Y is 6-10 membered aryl; X is 3-10 membered cycloalkyl; and m is an integer from 0-1

PCT publication number WO2004/110357 generically discloses a range of compounds, including certain 2-carboxy thiophene compounds, having phosphodiesterase 6 delta (PDE6D) modulating activity. The compounds disclosed have the formula (I)

wherein

R¹, R², R³, and R⁴ are independently selected from the group consisting of hydrogen, halogen, alkyl, cycloalkyl, aryl, heteroaryl, NR⁵C(O)R⁷, C(O)NR⁵R⁶, C(O)R⁷ and C(O)OR⁷, wherein R⁵, R⁶ and R⁷ are independently selected from hydrogen, lower alkyl, cycloalkyl or aryl, and where R⁵, R⁶, and R⁷ together can optionally form a 3, 4, 5, 6 or 7 membered ring optionally having one or more degrees of substitution.

Surprisingly, it has now been found that compounds according to the present invention, generically disclosed in WO2002/100851 , WO2003/062215, WO2004/052885 or WO2004/1 10357, and having a specific substitution pattern, exhibit improved properties over those compounds specifically disclosed in WO2002/100851 , WO2003/062215, WO2004/052885 or WO2004/110357.

SUMMARY OF THE INVENTION The present invention involves novel 2-carboxy thiophene compounds represented hereinbelow, pharmaceutical compositions comprising such compounds and use of the compounds in treating viral infection, especially HCV infection.

DETAILED DESCRIPTION OF THE INVENTION The present invention provides a compound of Formula (I) :

wherein:

A represents hydroxy; R¹ represents -R^x-R^γ;

R^x represents phenyl (optionally substituted at one of the mefø-positions to the thiophene by halo, methyl, ethyl or trifluoromethyl) or a 5- or 6-membered heteroaryl (optionally substituted at one of the mefø-positions to the thiophene by halo, methyl, ethyl, methoxy or trifluoromethyl) bonded through a ring carbon atom to the carbon atom of the thiophene;

R^γ represents a 8-, 9- or 10-membered heteroaryl, bonded such that when R^x is phenyl or a 6-membered heteroaryl, R^γ is in the para-position;

R² represents -C_5-7cycloalkyl optionally substituted by one or more substituents selected from fluoro, =CHCH₃, -OH, -OCH₃, =0 or -Ci_-6alkyl (wherein the alkyl may itself be optionally substituted by one or more fluoro atoms); with the proviso that when R¹ represents 4-imidazo[2,1-b][1 ,3]thiazol-6-ylphenyl, then R² may not represent trans-4- (trifluoromethyl)cyclohexyl;

R³ represents -(CH₂)₂-O-(CH₂)_mH, -CH-(CH₂OMe)₂, -CH(R⁴)-CH₂-O-(CH₂)_mH, - CH₂-CH(R⁴)- O-(CH₂)_mH, tetrahydrofuran-3-yl, -CH₂-tetrahydrofuran-3-yl, tetrahydro-2/-/-pyran-4-yl, -CH₂- tetrahydro-2/-/-pyran-4-yl or frans-4-hydroxycyclohex-1-yl;

R⁴ represents -Ci_-3alkyl;

m represents an integer selected from 1 or 2;

or salts, solvates or esters thereof.

The compounds of the present invention exhibit improved potency against the replication of HCV (1 a and 1 b genotypes), and therefore have the potential to achieve greater efficacy in man. High potency in both genotypes is considered to be advantageous.

There is provided as a further aspect of the present invention a compound of Formula (I) or pharmaceutically acceptable salts, solvates and esters thereof for use in human or veterinary medical therapy, particularly in the treatment or prophylaxis of viral infection, particularly flavivirus infection, for example HCV infection.

It will be appreciated that reference herein to therapy and/or treatment includes, but is not limited to prevention, retardation, prophylaxis, therapy and cure of the disease. It will further be appreciated that references herein to treatment or prophylaxis of HCV infection include treatment or prophylaxis of HCV-associated disease such as liver fibrosis, cirrhosis and hepatocellular carcinoma. In a further or alternative aspect there is provided a method for the treatment of a human or animal subject with viral infection, particularly HCV infection, which method comprises administering to said human or animal subject an effective amount of a compound of Formula (I) or pharmaceutically acceptable salts, solvates and esters thereof.

According to another aspect of the invention, there is provided the use of a compound of Formula (I) or pharmaceutically acceptable salts, solvates and esters thereof in the manufacture of a medicament for the treatment and/or prophylaxis of viral infection, particularly HCV infection.

It will be appreciated that the compounds of the present invention may contain one or more asymmetric carbon atoms and may exist in racemic, diastereoisomeric, and optically active forms. All of these racemic compounds, enantiomers and diastereoisomers are contemplated to be within the scope of the present invention.

In one aspect, R^x represents phenyl optionally substituted by halo, methyl or trifluoromethyl. In a further aspect, R^x represents unsubstituted phenyl.

In one aspect, R^γ represents furo[3,2-b]pyridin-2-yl, pyrazolo[1 ,5-a]pyrimidin-2-yl, imidazo[1 ,2-a]pyridin-2-yl, imidazo[2,1-t)][1 ,3]thiazol-6-yl, 7-amino-5-methylpyrazolo[1 ,5- a]pyrimidin-2-yl, 5-methylpyrazolo-[1 ,5-a]pyrimidin-2-yl, 7-aminopyrazolo[1 ,5-a]pyrimidin-2-yl, [1 ,3]oxazolo[4,5-b]pyridin-2-yl, furo[2,3-b]pyridin-5-yl, 5-amino-1 ,3-benzoxazol-2-yl, [1 ,3]oxazolo[5,4-b]pyridin-2-yl, or furo[3,2-c]pyridin-2-yl.

In a further aspect, R^γ represents furo[3,2-b]pyridin-2-yl, pyrazolo[1 ,5-a]pyrimidin-2-yl, imidazo[1 ,2-a]pyridin-2-yl, imidazo[2,1-t)][1 ,3]thiazol-6-yl or 7-aminopyrazolo[1 ,5-a]pyrimidin- 2-yl.

In a further aspect, R^γ represents furo[3,2-b]pyridin-2-yl, pyrazolo[1 ,5-a]pyrimidin-2-yl, imidazo[1 ,2-a]pyridin-2-yl, imidazo[2,1-b][1 ,3]thiazol-6-yl, 7-aminopyrazolo[1 ,5-a]pyrimidin-2- yl or [1 ,3]oxazolo[4,5-b]pyridin-2-yl.

In a yet further aspect, R^γ represents furo[3,2-b]pyridin-2-yl, pyrazolo[1 ,5-a]pyrimidin-2-yl, 7- aminopyrazolo[1 ,5-a]pyrimidin-2-yl or imidazo[1 ,2-a]pyridin-2-yl.

In one aspect, R² represents C₆cycloalkyl optionally substituted by one or more Ci_-4alkyl substituents (wherein the alkyl group may itself be optionally substituted by one or more fluoro atoms). In one aspect, the alkyl group is not substituted. In one aspect, R² represents Cβcycloalkyl substituted by methyl or trifluoromethyl. In one aspect, R² represents Cβcycloalkyl substituted at the 4-position, for example with trans stereochemistry. In a further aspect, R² represents frans-4-methylcyclohexyl or frans-4-(trifluoromethyl)cyclohexyl. In a further aspect, R² represents frans-4-methylcyclohexyl. In one aspect, R³ represents -(CH₂)₂-O-(CH₂)_mH, -CH-(CH₂OMe)₂, -CH(Me)CH₂-O-Me, tetrahydrofuran-3-yl, tetrahydro-2/-/-pyran-4-yl, -CH₂-tetrahydro-2/-/-pyran-4-yl or trans-4- hydroxycyclohex-1-yl. In a further aspect, R³ represents methoxyethyl, ethoxyethyl, 1- methyl-2-(methyloxy)ethyl, tetrahydrofuran-3-yl, tetrahydro-2/-/-pyran-4-yl or trans-4- hydroxycyclohex-1 -yl.

In a further aspect, R³ represents -(CH₂)₂-O-(CH₂)_mH, -CH-(CH₂OMe)₂, tetrahydrofuran-3-yl, tetrahydro-2/-/-pyran-4-yl or -CH₂-tetrahydro-2/-/-pyran-4-yl. In a further aspect, R³ represents methoxyethyl, ethoxyethyl, tetrahydrofuran-3-yl or tetrahydro-2/-/-pyran-4-yl. In a further aspect, R³ represents methoxyethyl, tetrahydrofuran-3-yl or tetrahydro-2/-/-pyran-4-yl.

In one aspect, R⁴ represents methyl.

In one aspect, R^x represents phenyl optionally substituted by halo, methyl or trifluoromethyl; R^γ represents furo[3,2-b]pyridin-2-yl, pyrazolo[1 ,5-a]pyrimidin-2-yl, imidazo[1 ,2-a]pyridin-2-yl, imidazo[2,1-t)][1 ,3]thiazol-6-yl, 7-amino-5-methylpyrazolo[1 ,5-a]pyrimidin-2-yl, 5- methylpyrazolo-[1 ,5-a]pyrimidin-2-yl, 7-aminopyrazolo[1 ,5-a]pyrimidin-2-yl, [1 ,3]oxazolo[4,5- b]pyridin-2-yl, furo[2,3-b]pyridin-5-yl, 5-amino-1 ,3-benzoxazol-2-yl, [1 ,3]oxazolo[5,4-b]pyridin- 2-yl, or furo[3,2-c]pyridin-2-yl; R² represents C₆cycloalkyl optionally substituted by one or more substituents (wherein the alkyl group may itself be optionally substituted by one or more fluoro atoms); and R³ represents -(CH₂)₂-O-(CH₂)_mH, -CH-(CH₂OMe)₂, - CH(Me)CH₂-O-Me, tetrahydrofuran-3-yl, tetrahydro-2/-/-pyran-4-yl, -CH₂-tetrahydro-2/-/-pyran- 4-yl or frans-4-hydroxycyclohexyl.

In a further aspect, R^x represents unsubstituted phenyl; R^γ represents furo[3,2-b]pyridin-2-yl, pyrazolo[1 ,5-a]pyrimidin-2-yl, imidazo[1 ,2-a]pyridin-2-yl, imidazo[2,1-b][1 ,3]thiazol-6-yl, 7- aminopyrazolo[1 ,5-a]pyrimidin-2-yl or [1 ,3]oxazolo[4,5-b]pyridin-2-yl; R² represents trans-4- methylcyclohexyl or frans-4-(trifluoromethyl)cyclohexyl; and R³ represents methoxyethyl, ethoxyethyl, 1-methyl-2-(methyloxy)ethyl, tetrahydrofuran-3-yl, tetrahydro-2/-/-pyran-4-yl or frans-4-hydroxycyclohexyl.

In one aspect, the present invention provides at least one chemical entity chosen from compounds of Formula (Ia) :

wherein: A represents hydroxy;

R¹ represents -R^x-R^γ;

R^x represents phenyl (optionally substituted by halo, methyl, ethyl, methoxy or trifluoromethyl) or a 5- or 6-membered heteroaryl bonded through a ring carbon atom to the carbon atom of the thiophene;

R^γ represents a 8-, 9- or 10-membered heteroaryl, bonded such that when R^x is phenyl, R^γ is in the para-position;

R² represents C₆cycloalkyl optionally substituted by one or more unsubstituted Ci_-4alkyl substituents;

R³ represents -(CH₂)₂-O-(CH₂)_mH, -CH-(CH₂OMe)₂, tetrahydrofuran-3-yl, tetrahydro-2H- pyran-4-yl or -CH₂- tetrahydro-2/-/-pyran-4-yl;

m represents an integer selected from 1 or 2;

or salts, solvates or esters thereof.

In one aspect, the present invention provides at least one chemical entity chosen from compounds of Formula (Ib) :

wherein:

A represents hydroxy;

R¹ represents -R^x-R^γ;

R^x represents phenyl (optionally substituted by halo, methyl, ethyl, methoxy or trifluoromethyl) or a 5 or 6-membered heteroaryl bonded through a ring carbon atom to the carbon atom of the thiophene;

R^γ represents a 8, 9 or 10-membered heteroaryl, bonded such that when R^x is phenyl or a 6- membered heteroaryl, R^γ is in the para-position; R² represents C_5-7cycloalkyl optionally substituted by one or more substituents selected from fluoro, =CHCH₃, OH, OCH₃, =0 or -Ci_-6alkyl (wherein the alkyl may itself be optionally substituted by one or more fluoro atoms); with the proviso that when R¹ represents 4-imidazo[2,1-b][1 ,3]thiazol-6-ylphenyl, then R² may not represent trans-4- (trifluoromethyl)cyclohexyl;

R³ represents -(CH₂)₂-O-(CH₂)_mH, -CH-(CH₂OMe)₂, -CH(R⁴)-CH₂-O-(CH₂)_mH, - CH₂-CH(R⁴)- O-(CH₂)_mH , tetrahydrofuran-3-yl, tetrahydro-2/-/-pyran-4-yl or -CH₂-tetrahydro-2/-/-pyran-4-yl;

R⁴ represents Ci_-3alkyl

m represents an integer selected from 1 or 2;

and salts, solvates and esters thereof.

As used herein the term "at least one chemical entity" means at least one chemical substance chosen from the group of compounds consisting of compounds of Formula I and pharmaceutically acceptable derivatives thereof.

It is to be understood that the present invention covers all combinations of aspects, suitable, convenient and preferred groups described herein.

As used herein, the term "compounds of the invention" means the compounds according to Formula I and the salts, solvates and esters thereof. The term "a compound of the invention" means any one of the compounds of the invention as defined above.

As used herein the term "at least one chemical entity" means at least one chemical substance chosen from the group of compounds consisting of compounds of Formula I and salts, solvates and esters thereof.

As used herein, "acetyl" refers to -C(O)CH₃.

As used herein unless otherwise specified, "alkyl" refers to an optionally substituted hydrocarbon group. The alkyl hydrocarbon group may be linear, branched or cyclic, saturated or unsaturated. Where the alkyl group is linear or branched, examples of such groups include methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl or hexyl and the like. Where the alkyl hydrocarbon group is unsaturated, it will be understood that there will be a minimum of 2 carbon atoms in the group, for example an alkenyl or alkynyl group. Where the alkyl hydrocarbon group is cyclic, it will be understood that there will be a minimum of 3 carbon atoms in the group. In one aspect, alkyl moieties are saturated. In one aspect, alkyl moieties are -Ci_-4alkyl. Unless otherwise stated, optional substituents include -Ci_-6alkyl (unsubstituted), =CH(CH₂)_tH, fluoro, -CF₃, -OR^E, -SR^E, -C(O)NR^BR^C, -C(O)R⁰, -CO₂H, -CO₂R⁰, -NR^BR^C, -NR^AC(O)R°, -NR^ACO₂R^D, -NR^AC(O)NR^FR^G, -SO₂NR^FR^G, -SO₂R⁰, nitro, cyano, oxo, aryl, heteroaryl and heterocyclyl.

As used herein, the term "alkenyl" refers to a linear or branched hydrocarbon group containing one or more carbon-carbon double bonds. In one aspect, the alkenyl group has from 2 to 6 carbon atoms. Examples of such groups include ethenyl, propenyl, butenyl, pentenyl or hexenyl and the like.

As used herein, the term "alkynyl" refers to a linear or branched hydrocarbon group containing one or more carbon-carbon triple bonds. In one aspect the alkynyl group has from 2 to 6 carbon atoms. Examples of such groups include ethynyl, propynyl, butynyl, pentynyl or hexynyl and the like.

As used herein unless otherwise specified, "cycloalkyl" refers to an optionally substituted, cyclic hydrocarbon group. The hydrocarbon group may be saturated or unsaturated, monocyclic or bridged bicyclic. Where the cycloalkyl group is saturated, examples of such groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl and the like. Where the cycloalkyl group is unsaturated, examples of such groups include cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl or cyclooctenyl and the like. In one aspect, the cycloalkyl group has from 5 to 7 carbon atoms. In one aspect, cycloalkyl moieties are cyclohexenyl, cyclopentenyl and cyclohexyl. Unless otherwise stated, the cycloalkyl group may be substituted by one or more optional substituents including -Ci_-6alkyl (unsubstituted), =CH(CH₂)_tH, fluoro, -CF₃, -OR^E, -SR^E, -C(O)NR^BR^C, -C(O)R⁰, -CO₂H, -CO₂R⁰, -NR^BR^C, -NR^AC(0)R°, -NR^AC0₂R°, -NR^AC(0)NR^FR^G, -SO₂NR^FR^G, -SO₂R⁰, nitro, cyano, oxo, phenyl and heterocyclyl.

As used herein, the term "alkoxy" refers to an -O-alkyl group wherein alkyl is as defined herein. Examples of such groups include methoxy, ethoxy, propoxy, butoxy, pentoxy or hexoxy and the like.

As used herein, "aryl" refers to an optionally substituted aromatic group with at least one ring having a conjugated pi-electron system, containing up to two conjugated or fused ring systems. "Aryl" includes carbocyclic aryl and biaryl groups, all of which may be optionally substituted. In one aspect, "aryl" moieties contain 6-10 carbon atoms. In one aspect, "aryl" moieties are unsubstituted, monosubstituted, disubstituted or trisubstituted phenyl. In one aspect, unless otherwise stated, "aryl" substituents are selected from the group consisting of -Ci_-6alkyl, halo, -OR^E, -SR^E, -C(O)NR^BR^C, -C(O)R⁰, -CO₂H, -CO₂R⁰, -NR^BR^C, -NR^AC(0)R°, -NR^AC0₂R°, -NR^AC(0)NR^FR^G, -SO₂NR^FR^G, -SO₂R⁰, nitro, cyano, heterocyclyl, -CF₃, -OCF₃ and phenyl.

As used herein, "carbonyl" refers to -C(O)-. As used herein, "cyano" refers to -CN.

As used herein, "halogen" or "halo" refer to a fluorine, chlorine, bromine or iodine atom. References to "fluoro", "chloro", "bromo" or "iodo" should be construed accordingly.

As used herein, unless otherwise specified, "heteroaryl" refers to an optionally substituted, 5- , 6-, 8-, 9- or 10-membered, aromatic group comprising one to four heteroatoms selected from N, O and S, with at least one ring having a conjugated pi-electron system, containing up to two conjugated or fused ring systems. In one aspect, "heteroaryl" moieties are unsubstituted, monosubstituted, disubstituted or trisubstituted (where applicable) pyridine, pyrazine, thiazole, thiophene, oxadiazole, oxazole, pyrimidine, pyridazine, benzodioxole, benzofuran, benzodioxin, indole, benzimidazole, benzofuran, indole, indazole, isoindole, benzothiophene, benzothiazole, benzoxazole, benzisoxazole, benzisothiazole, benzotriazole, furopyridine, furopyrimidine, furopyridazine, furopyrazine, furotriazine, pyrrolopyridine, pyrrolopyrimidine, pyrrolopyridazine, pyrrolopyrazine, pyrrolotriazine, thienopyridine, thienopyrimidine, thienopyridazine, thienopyrazine, thienotriazine, thiazolopyridine, thiazolopyrimidine, thiazolopyridazine, thiazolopyrazine, thiazolotriazine, oxazolopyridine, oxazolopyrimidine, oxazolopyridazine, oxazolopyrazine, oxazolotriazine, imidazopyridine, imidazopyrimidine, imidazopyridazine, imidazopyrazine, imidazotriazine, pyrazolopyridine, pyrazolopyrimidine, pyrazolopyridazine, pyrazolopyrazine, pyrazolotriazine, triazolopyridine, triazolopyrimidine, triazolopyridazine, triazolopyrazine, quinoline, naphthyridine, quinoxaline, quinazoline, isoquinoline, cinnoline, pyridopyridazine, pyridopyrimidine, pyridopyrazine, pyrazinopyrazine, pteridine, pyrazinopyridazine, pyrimidopyridazine, pyrimidopyrimidine, imidazothiazole, thiazolooxazole. All isomers of the above heteroaryls are within the scope of this invention. Each heteroaryl group may be attached at any ring carbon or may be attached through nitrogen when the nitrogen is part of a 5-membered ring. In one aspect, unless otherwise stated, "heteroaryl" substituents are selected from the group consisting of -Ci_-6alkyl, halo, -OR^E, -SR^E, -C(O)NR^BR^C, -C(O)R⁰, -CO₂R⁰, -NR^BR^C, -NR^AC(O)R°, -NR^ACO₂R^D, -NR^AC(O)NR^FR^G, -SO₂NR^FR^G, -SO₂R⁰, oxo, nitro, cyano, heterocyclyl, -CF₃ and phenyl.

As used herein, "heterocyclic" and "heterocyclyl" refer to an optionally substituted, 5- or 6- membered, saturated or partially saturated, cyclic group containing 1 or 2 heteroatoms selected from N, optionally substituted by hydrogen, -Ci_-6alkyl, -C(O)R⁰, -C(O)NR^BR^C, -C(O)OH, -SO₂R⁰, aryl or heteroaryl; O; and S, optionally substituted by one or two oxygen atoms. Ring carbon atoms may be optionally substituted by -Ci-βalkyl, -0R^A, -C(O)R⁰, or -SO₂R⁰. In one aspect, unless otherwise stated, "heterocyclic" moieties are unsubstituted or monosubstituted tetrahydro-2H-pyran-4-yl, piperidinyl and tetrahydrofuran- 3-yl.

As used herein, "nitro" refers to -NO₂. As used herein, "oxo" refers to =0.

As used herein, "Et" refers to "ethyl", "iPr" refers to "isopropyl", "Me" refers to "methyl", "OBn" refers to "benzyloxy", and "Ph" refers to "phenyl".

R^A represents hydrogen or -Ci_-6alkyl.

R^B and R^c independently represent hydrogen, -d-βalkyl, aryl, heterocyclyl or heteroaryl; or RR^BB aanndd RR^cc ttooggeetthheerr wwiitthh tthhee nniittirogen atom to which they are attached form a 5 or 6 membered saturated cyclic group.

R^D is selected from the group consisting of -Ci_-6alkyl, aryl, heterocyclyl, heteroaryl, arylalkyl, and heteroarylalkyl.

R^E represents hydrogen, -Ci_-6alkyl, arylalkyl, heteroarylalkyl, aryl, heterocyclyl or heteroaryl.

R^F and R^G are independently selected from the group consisting of hydrogen, -d-βalkyl, aryl, heteroaryl, arylalkyl, and heteroarylalkyl; or R^F and R^G together with the nitrogen atom to which they are attached form a 5- or 6-membered saturated cyclic group.

In a further aspect, the present invention provides a compound chosen from the group consisting of:

3-[[(trans-4-methylcyclohexyl)carbonyl](tetrahydro-3-furanyl)amino]-5-(4-pyrazolo[1 ,5- a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylic acid;

3-[[(trans-4-methylcyclohexyl)carbonyl](tetrahydro-2H-pyran-4-yl)amino]-5-(4-pyrazolo[1 ,5- a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylic acid;

5-(4-furo[3,2-b]pyridin-2-ylphenyl)-3-[[(trans-4-methylcyclohexyl)carbonyl](tetrahydro-2H- pyran-4-yl)amino]-2-thiophenecarboxylic acid; 5-[4-(7-aminopyrazolo[1 ,5-a]pyrimidin-2-yl)phenyl]-3-[[(trans-4- methylcyclohexyl)carbonyl](tetrahydro-2H-pyran-4-yl)amino]-2-thiophenecarboxylic acid;

5-(4-imidazo[1 ,2-a]pyridin-2-ylphenyl)-3-[[(trans-4-methylcyclohexyl)carbonyl](tetrahydro-2H- pyran-4-yl)amino]-2-thiophenecarboxylic acid;

5-(4-imidazo[2,1-b][1 ,3]thiazol-6-ylphenyl)-3-[[(trans-4-methylcyclohexyl)carbonyl](tetrahydro- 2H-pyran-4-yl)amino]-2-thiophenecarboxylic acid;

5-(4-furo[3,2-b]pyridin-2-ylphenyl)-3-{[(trans-4-methylcyclohexyl)carbonyl][2-

(methyloxy)ethyl]amino}-2-thiophenecarboxylic acid;

5-[4-(7-aminopyrazolo[1 ,5-a]pyrimidin-2-yl)phenyl]-3-{[(trans-4-methylcyclohexyl)carbonyl][2-

(methyloxy)ethyl]amino}-2-thiophenecarboxylic acid; 3-{[(trans-4-methylcyclohexyl)carbonyl][2-(methyloxy)ethyl]amino}-5-(4-pyrazolo[1 ,5- a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylic acid;

5-(4-imidazo[1 ,2-a]pyridin-2-ylphenyl)-3-{[(trans-4-methylcyclohexyl)carbonyl][2-

(methyloxy)ethyl]amino}-2-thiophenecarboxylic acid; 5-(4-imidazo[2,1-b][1 ,3]thiazol-6-ylphenyl)-3-{[(trans-4-methylcyclohexyl)carbonyl][2-

(methyloxy)ethyl]amino}-2-thiophenecarboxylic acid;

3-{[2-(ethyloxy)ethyl][(trans-4-methylcyclohexyl)carbonyl]amino}-5-(4-furo[3,2-b]pyridin-2- ylphenyl)-2-thiophenecarboxylic acid;

5-[4-(7-aminopyrazolo[1 ,5-a]pyrimidin-2-yl)phenyl]-3-{[2-(ethyloxy)ethyl][(trans-4- methylcyclohexyl)carbonyl]amino}-2-thiophenecarboxylic acid;

3-{[2-(ethyloxy)ethyl][(trans-4-methylcyclohexyl)carbonyl]amino}-5-(4-pyrazolo[1 ,5- ajpyrimidin^-ylphenyl^-thiophenecarboxylic acid;

3-{[2-(ethyloxy)ethyl][(trans-4-methylcyclohexyl)carbonyl]amino}-5-(4-imidazo[1 ,2-a]pyridin-2- ylphenyl)-2-thiophenecarboxylic acid; 3-{[2-(ethyloxy)ethyl][(trans-4-methylcyclohexyl)carbonyl]amino}-5-(4-imidazo[2, 1 - b][1 ,3]thiazol-6-ylphenyl)-2-thiophenecarboxylic acid;

3-([(trans-4-methylcyclohexyl)carbonyl]{2-(methyloxy)-1-[(methyloxy)methyl]ethyl}amino)-5-

(4-pyrazolo[1 ,5-a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylic acid;

5-(4-furo[3,2-b]pyridin-2-ylphenyl)-3-[[(trans-4-methylcyclohexyl)carbonyl](tetrahydro-3- furanyl)amino]-2-thiophenecarboxylic acid;

5-(4-imidazo[1 ,2-a]pyridin-2-ylphenyl)-3-[[(trans-4-methylcyclohexyl)carbonyl](tetrahydro-2H- pyran-4-ylmethyl)amino]-2-thiophenecarboxylic acid;

5-(4-furo[3,2-b]pyridin-2-ylphenyl)-3-[[(trans-4-methylcyclohexyl)carbonyl](tetrahydro-2H- pyran-4-ylmethyl)amino]-2-thiophenecarboxylic acid; 3-[[(trans-4-methylcyclohexyl)carbonyl](tetrahydro-2H-pyran-4-ylmethyl)amino]-5-(4- pyrazoloπ .δ-ajpyrimidin^-ylphenyl^-thiophenecarboxylic acid;

3-([2-(methyloxy)ethyl]{[trans-4-(trifluoromethyl)cyclohexyl]carbonyl}amino)-5-(4- pyrazoloπ .δ-alpyrimidin^-ylphenyl^-thiophenecarboxylic acid;

5-(4-furo[3,2-b]pyridin-2-ylphenyl)-3-([2-(methyloxy)ethyl]{[trans-4- (trifluoromethyl)cyclohexyl]carbonyl}amino)-2-thiophenecarboxylic acid;

5-(4-imidazo[2,1-b][1 ,3]thiazol-6-ylphenyl)-3-[[(trans-4-methylcyclohexyl)carbonyl](tetrahydro-

2H-pyran-4-ylmethyl)amino]-2-thiophenecarboxylic acid;

5-(4-pyrazolo[1 ,5-a]pyrimidin-2-ylphenyl)-3-(tetrahydro-2H-pyran-4-yl{[trans-4-

(trifluoromethyl)cyclohexyl]carbonyl}amino)-2-thiophenecarboxylic acid; 5-(4-furo[3,2-b]pyridin-2-ylphenyl)-3-(tetrahydro-2H-pyran-4-yl{[trans-4-

(trifluoromethyl)cyclohexyl]carbonyl}amino)-2-thiophenecarboxylic acid formic acid salt; 5-[4-(7-aminopyrazolo[1 ,5-a]pyrimidin-2-yl)phenyl]-3-(tetrahydro-2H-pyran-4-yl{[trans-4- (trifluoromethyl)cyclohexyl]carbonyl}amino)-2-thiophenecarboxylic acid; 3-[[(trans-4-methylcyclohexyl)carbonyl](tetrahydro-2H-pyran-4-ylmethyl)amino]-5-(4- [i ^oxazolo^S-blpyridin^-ylphenyl^-thiophenecarboxylic acid; 3-{[(trans-4-methylcyclohexyl)carbonyl][1-methyl-2-(methyloxy)ethyl]amino}-5-(4- pyrazoloπ .δ-ajpyrimidin^-ylphenyl^-thiophenecarboxylic acid; 5-(4-imidazo[1 ,2-a]pyridin-2-ylphenyl)-3-(tetrahydro-2H-pyran-4-yl{[trans-4- (trifluoromethyl)cyclohexyl]carbonyl}amino)-2-thiophenecarboxylic acid; 3-{(frans-4-hydroxycyclohexyl)[(frans-4-methylcyclohexyl)carbonyl]amino}-5-(4-pyrazolo[1 ,5- a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylic acid;

and salts, solvates and esters, and individual enantiomers thereof where appropriate.

Also included in the present invention are pharmaceutically acceptable salt complexes. The present invention also covers the pharmaceutically acceptable salts of the compounds of

Formula (I). Suitable pharmaceutically acceptable salts of the compounds of Formula (I) include acid salts, for example sodium, potassium, calcium, magnesium and tetraalkylammonium and the like, or mono- or di- basic salts with the appropriate acid for example organic carboxylic acids such as acetic, lactic, tartaric, malic, isethionic, lactobionic and succinic acids; organic sulfonic acids such as methanesulfonic, ethanesulfonic, benzenesulfonic and p-toluenesulfonic acids and inorganic acids such as hydrochloric, sulfuric, phosphoric and sulfamic acids and the like.

The present invention also relates to solvates of the compounds of Formula (I), for example hydrates.

The present invention also relates to pharmaceutically acceptable esters of the compounds of Formula (I), for example carboxylic acid esters -COOR, in which R is selected from straight or branched chain alkyl, for example n-propyl, n-butyl, alkoxyalkyl (e.g. methoxymethyl), aralkyl (e.g. benzyl), aryloxyalkyl (e.g. phenoxymethyl), aryl (e.g. phenyl optionally substituted by halogen,

or amino); or for example -CH₂OC(O)R' or -CH₂OCO₂R' in which R' is alkyl (e.g. R' is f-butyl). Unless otherwise specified, any alkyl moiety present in such esters preferably contains 1 to 18 carbon atoms, particularly 1 to 4 carbon atoms. Any aryl moiety present in such esters preferably comprises a phenyl group.

In one aspect, the compound of Formula (I) is in the form of parent compound, a salt or a solvate.

As used herein, the term "pharmaceutically acceptable" used in relation to an ingredient (active ingredient such as an active ingredient, a salt thereof or an excipient) which may be included in a pharmaceutical formulation for administration to a patient, refers to that ingredient being acceptable in the sense of being compatible with any other ingredients present in the pharmaceutical formulation and not being deleterious to the recipient thereof.

Throughout the specification and the claims which follow, unless the context requires otherwise, the word 'comprise', and variations such as 'comprises' and 'comprising', will be understood to imply the inclusion of a stated integer or step or group of integers but not to the exclusion of any other integer or step or group of integers or steps.

It will further be appreciated that certain compounds of the present invention may exist in different tautomeric forms. All tautomers are contemplated to be within the scope of the present invention.

PROCESSES

Compounds of Formula (I) in which A is hydroxy may be prepared from a compound of Formula (II)

in which A is a protected hydroxy group, for example an alkoxy, benzyloxy or silyloxy group and R¹, R², and R³ are as defined above for Formula (I). For example when A is methoxy or ethoxy, and R¹, R², R³ and R⁴ are as defined above for Formula (I), by treatment with an appropriate base, for example aqueous sodium hydroxide or lithium hydroxide, optionally in a solvent such as methanol, tetrahydrofuran or combinations thereof. Suitably, the temperature is in the range 25 to 100⁰C, for example 25 to 5O⁰C. Alternatively, when A is methoxy or ethoxy and R¹, R² and R³ are as defined above for Formula (I), by treatment with lithium iodide in a suitable solvent such as pyridine, lutidine or collidine, suitably in the temperature range 100-170⁰C.

For example when A is te/f-butoxy, and R¹, R² and R³ are as defined above for Formula (I), by treatment with an appropriate acid, for example trifluoroacetic acid. Suitably, the reaction is carried out in a solvent, for example dichloromethane. Suitably, the temperature is in the range 0 to 5O⁰C, for example 15 to 3O⁰C.

For example when A is silyloxy, and R¹, R² and R³ and are as defined above for Formula (I), by treatment with a suitable fluoride source for example tetrabutylammonium fluoride. The reaction is carried out in a suitable solvent, for example tetrahydrofuran. Suitably, the temperature is in the range 0 to 5O⁰C, for example 15 to 3O⁰C. Compounds of Formula (I) in which A is hydroxy, or (II) in which A is an alkoxy, benzyloxy or silyloxy group and R¹ and R³ are as defined above for Formula (I), may be prepared by reaction of a compound of

in which A is hydroxy or an alkoxy, benzyloxy or silyloxy group, and R² and R³ are as defined above for Formula (I) and X is a halogen such as bromide or iodide; with a suitable boronic acid R¹-B(OH)₂ or boronate ester R¹-B(0R')(0R"), in which R' and R" are independently alkyl or R' and R" together with the carbon atoms to which they are attached form a ring optionally substituted by alkyl, such as a pinacol ester, in the presence of a palladium catalyst such as tetrakistriphenyl phosphine palladium or bis-[(diphenylphosphino)- ferrocene]-palladium-(ll) chloride, in the presence of a suitable base such as sodium carbonate, in a suitable solvent such as DMF, DME, 1 ,4-dioxane, methanol or toluene, or combinations thereof, at a temperature in the range 50-100⁰C, optionally under an inert atmosphere.

Compounds of Formula (I) in which A is hydroxy, or (II) in which A is an alkoxy, benzyloxy or silyloxy group and R¹, R² and R³ are as defined above for Formula (I), may be prepared by reaction of a compound of For

in which A is hydroxy or an alkoxy, benzyloxy or silyloxy group, and R² and R³ are as defined above for Formula (I) and X is a suitable boronic acid -B(OH)₂ or boronate ester -B(OR')(OR"), in which R' and R" are independently alkyl or R' and R" together with the carbon atoms to which they are attached form a ring optionally substituted by alkyl, such as a pinacol ester, with R¹-Hal wherein Hal is a halogen such as bromide or iodide, in the presence of a palladium catalyst such as (tetrakistriphenyl phosphine) palladium or bis- [(diphenylphosphino)-ferrocene]-palladium-(ll) chloride, in the presence of a suitable base such as sodium carbonate, in a suitable solvent such as DMF, DME, 1 ,4-dioxane, methanol or toluene, or combinations thereof, at a temperature in the range 50-100⁰C, optionally under an inert atmosphere.

Compounds of Formula (III) in which A is an alkoxy, benzyloxy or silyloxy group may be prepared from compounds of Formula (IV)

in which A is an alkoxy, benzyloxy or silyloxy, and R ^(I2V a^>nd R³ are as defined above for Formula (I), by treatment with a suitable base such as lithium diisopropylamide and a halogen source such as iodine in a suitable solvent such as tetrahydrofuran, and at a temperature in the range -78°C to -20⁰C, optionally under an inert atmosphere.

Compounds of Formula (III)' in which A is an alkoxy, benzyloxy or silyloxy group may be prepared from compounds of

in which A is an alkoxy, benzyloxy or silyloxy, and R ^(I2V a^>nd R³ are as defined above for Formula (I), by treatment with a suitable base such as lithium diisopropylamide and a boronate source such as B(OR)3 wherein R is an alkyl group, for example methyl, in a suitable solvent such as tetrahydrofuran, and at a temperature in the range -78°C to -20⁰C.

Compounds of Formula (III) in which A is hydroxy may be prepared from compounds of Formula (III) in which A is an alkoxy, benyloxy or silyloxy group, for example by treatment with an appropriate base, acid or fluoride source as described in relation to the preparation of compounds of Formula (I) from compounds of Formula (II).

Compounds of Formula (IV) in which A is an alkoxy, benzyloxy or silyloxy group and R² and

R are as defined above may be prepared by reaction of a compound of Formula (V)

in which A an alkoxy, benzyloxy or silyloxy group, and R¹ and R³ are as defined above for Formula (I); with a suitable acylating agent, for example R²-C(O)-Y, wherein Y is a halo atom, for example chloro or bromo, and R² is as defined above for Formula (I). The reaction may be carried out in a suitable solvent, for example dichloromethane, and optionally in the presence of a suitable base, for example pyridine or triethylamine. A phosphine such as triphenylphosphine may optionally be used in place of the base. Compounds of Formula (V) in which A is an alkoxy, benzyloxy or silyloxy group may be prepared by reaction of a compound of Formula (Vl)

in which A an alkoxy, benzyloxy or silyloxy group, by treatment with a suitable vinyl ether, or a suitable aldehyde or a suitable ketone in the presence of a suitable acid, such as acetic acid, and a suitable reducing agent such as sodium triacetoxyborohydride, in a suitable solvent such as dichloromethane. Alternatively, compounds of Formula (V) in which A is an alkoxy, benzyloxy or silyloxy group may be prepared from compounds of Formula (Vl) in which A is an alkoxy, benzyloxy or silyloxy are as defined above for Formula (I), by treatment with a suitable alkylating agent R³-X' where X' is a halo group such as chloride, bromide or iodide, or X' is a sulphonate ester such as methanesulfonate and R³ is as defined above for Formula (I), in suitable solvent such as dimethylformamide in the presence of a suitable base such as triethylamine.

Compounds of Formula (V) may also be prepared by reacting a compound of Formula (VII)

in which A is an alkoxy, benzyloxy or silyloxy group and X is a halo group such as bromo, with an amine R³-NH₂ in the presence of a palladium catalyst such as tris(dibenzylidenacetone)dipalladium in the presence of a reagent such as 2,2'- bis(diphenylphosphino)-1 ,1 '-binaphthyl (BINAP) and a base such as cesium carbonate, in a suitable solvent such as toluene and in a suitable temperature range such as 80-120⁰C.

Compounds of Formula (IV) in which A is an alkoxy, benzyloxy or silyloxy group and R² and R³ are as defined above for Formula (I), may also be prepared by reaction of a compound of Formula (VIII)

in which A is an alkoxy, benzyloxy or silyloxy group, and R² is as defined above for Formula

(I); with a suitable alkylating agent R³-X' in which X' is a halo atom such as chloro, bromo or iodo, or X' is a sulphonate ester such as methanesulfonate and R³ is as defined above for Formula (I), in a suitable solvent such as dimethylformamide, in the presence of a suitable base such as triethylamine and/or sodium hydride.

Compounds of Formula (VIII) in which A is an alkoxy, benzyloxy or silyloxy group may be prepared by reaction of a compound of Formula (Vl) in which A an alkoxy, benzyloxy or silyloxy group, with a suitable acylating agent, for example R²-C(O)-Y, wherein Y is a halo atom, for example chloro or bromo, and R² is as defined above for Formula (I). The reaction may be carried out in a suitable solvent, for example dichloromethane, in the presence of a suitable base, for example pyridine or triethylamine. A phosphine such as triphenylphosphine may optionally be used in place of the base.

Compounds of Formula (II) in which A is an alkoxy, benzyloxy or silyloxy group may also be prepared by reaction of a com

in which A an alkoxy, benzyloxy or silyloxy group, and R¹ and R³ are as defined above for Formula (I) with a suitable acylating agent, for example R²-C(0)-Y, wherein Y is a halo atom, for example chloro or bromo, and R² is as defined above for Formula (I). The reaction may be carried out in a suitable solvent, for example dichloromethane, in the presence of a suitable base, for example pyridine or triethylamine and thereafter removing any protecting group if desired. A phosphine such as triphenylphosphine may optionally be used in place of the base.

Compounds of Formula (IX) in which A is an alkoxy, benzyloxy or silyloxy group and R¹ and R³ are as defined above for Formula (I), may also be prepared by reaction of a compound of Formula (X)

in which X is a halogen such as bromide or iodide and R³ is as defined above for Formula (I), with a suitable boronic acid R¹-B(OH)₂ or boronate ester R¹-B(OR')₂ wherein R¹ is as defined above for Formula (I), in the presence of a palladium catalyst such as tetrakistriphenyl phosphine palladium or bis-[(diphenylphosphino)-ferrocene]-palladium-(ll) chloride, in the presence of a suitable base such as sodium carbonate, in a suitable solvent such as DMF, methanol or toluene, or combinations thereof, at a temperature in the range 50-100⁰C, optionally under an inert atmosphere. Compounds of Formula (X) in which A is an alkoxy, benzyloxy or silyloxy group, R³ is as defined above for Formula (I) and X is a halogen such as bromide or iodide, may be prepared by reaction of a compound of Formula (Xl)

wherein A is an alkoxy, benzyloxy or silyloxy group and X is a halogen such as bromide or iodide, by treatment with a suitable vinyl ether, or a suitable aldehyde or a suitable ketone in the presence of a suitable acid, such as acetic acid, and a suitable reducing agent such as sodium triacetoxyborohydride, in a suitable solvent such as dichloromethane.

Alternatively, compounds of Formula (X) in which A is an alkoxy, benzyloxy or silyloxy group may be prepared from compounds of Formula (Xl) in which A is an alkoxy, benzyloxy or silyloxy, by treatment with a suitable alkylating agent R³-X' where X' is a halo group such as chloride, bromide or iodide, or X' is a sulphonate ester such as methanesulfonate and R² is as defined above for Formula (I), in suitable solvent such as dimethylformamide in the presence of a suitable base such as triethylamine.

Compounds of Formula (X) in which A is an alkoxy, benzyloxy or silyloxy group, R³ is as defined above for Formula (I) and X is a halogen such as bromide or iodide, may also be prepared by reaction of a compound of Formula (V), by treatment with a suitable base such as lithium diisopropylamide and a halogen source such as iodine or bromine, in a suitable solvent such as tetrahydrofuran, and at a temperature in the range -78°C to -20⁰C.

Compounds of Formula (Xl) in which A is an alkoxy, benzyloxy or silyloxy group as defined above for Formula (I) and X is a halogen such as iodide, may be prepared from a compound of Formula (XII)

in which P is a suitable protecting group such as COCF₃ or CO₂ ¹Bu. For example, when P is COCF₃, by treatment with a suitable base such as aqueous potassium carbonate optionally in the presence of an alcohol such as methanol, or when P is CO₂ ¹Bu by treatment with a suitable acid such as hydrochloric acid in a suitable solvent such as 1 ,4-dioxane.

Compounds of Formula (XII) in which A is an alkoxy, benzyloxy or silyloxy group as defined above for Formula (I), X is a halo atom such as iodide and P is a suitable protecting group such as COCF₃ or CO₂ ¹Bu, may be prepared by reaction of a compound of Formula (XIII)

wherein P is a suitable protecting group such as COCF₃ or CO₂ ¹Bu, with a suitable base such as lithium diisopropylamide and a halogen source such as iodine, in a suitable solvent such as tetrahydrofuran, and at a temperature in the range -78°C to -20⁰C.

Compounds of Formula (XIII) in which A is an alkoxy, benzyloxy or silyloxy group as defined above for Formula (I) may be prepared by treating compounds of Formula (VI) with trifluoroacetic anhydride or di-tert-butyl di carbonate in a suitable solvent such as ether or acetonitrile, optionally in the presence of a catalyst such as DMAP.

Compounds of Formula (III) in which A is an alkoxy, benzyloxy or silyloxy group may also be prepared from compounds of Formula (XIV)

in which A is an alkoxy, benzyloxy or silyloxy, and R² is as defined above for Formula (I), by treatment with a suitable alkylating agent R³-X' where X' is a halo group such as chloride, bromide or iodide, or X' is a sulphonate ester such as methanesulfonate and R³ is as defined above for Formula (I), in suitable solvent such as dimethylformamide in the presence of a suitable base such as triethylamine.

Compounds of Formula (XIV) in which A is an alkoxy, benzyloxy or silyloxy group may also be prepared from compounds of Formula (Xl) with a suitable acylating agent, for example R²- C(O)-Y, wherein Y is a halo atom, for example chloro or bromo, and R² is as defined above for Formula (I). The reaction may be carried out in a suitable solvent, for example dichloromethane, in the presence of a suitable base, for example pyridine or triethylamine. A phosphine such as triphenylphosphine may optionally be used in place of the base.

Compounds of Formula (Vl) and (VII) are commercially available or well known in the art.

Compounds of Formula (I) in which A is hydroxy, or (II) in which A is an alkoxy, benzyloxy or silyloxy group, may be prepared by reaction of a compound of Formula (II)'

in which Z represents a halo substituent, and R^x, R², R³, and A are as defined for Formula

(II), by reaction with a suitable heteroaryl boronic acid, R^γ-boronic acid in which R^γ is as defined in Formula (I), in the presence of a palladium catalyst such as palladium (II) acetate, Pd(PPh₃)₄, a reagent such as 2-dicyclohexylphosphino-2'(N,N-dimethylamino)-biphenyl, and an additional reagent such as caesium fluoride, in a suitable solvent such as 1 ,4-dioxane.

Alternatively, the heteroaryl boronic acids may be reacted in the presence of a palladium catalyst such as palladium (II) acetate, a reagent such as 2-dicyclohexylphosphino-2'(N,N- dimethylamino)-biphenyl, and an additional reagent such as caesium fluoride, in a suitable solvent such as 1 ,4-dioxane.

Compounds of Formula (I) in which A is hydroxy, or (II) in which A is an alkoxy, benzyloxy or silyloxy group, may also be prepared by reaction of a compound of Formula (II)'

in which Z represents B(OH)₂, and R^x, R², R³ and A are as defined for Formula (II), by reaction with a suitable heteroaryl halide R^γ-hal, in which suitably the halide is bromo or iodo, in the presence of a palladium catalyst such as palladium (II) acetate, Pd(PPh₃)₄, a reagent such as 2-dicyclohexylphosphino-2'(N,N-dimethylamino)-biphenyl, and an additional reagent such as caesium fluoride, in a suitable solvent such as 1 ,4-dioxane. Alternatively, the heteroaryl boronic acids may be reacted in the presence of a palladium catalyst such as palladium (II) acetate, a reagent such as 2-dicyclohexylphosphino-2'(N,N-dimethylamino)- biphenyl, and an additional reagent such as caesium fluoride, in a suitable solvent such as 1 ,4-dioxane.

Compounds of Formula (II)' in which Z is halo may be prepared by reaction of a compound of Formula (III) with a boronic acid of Formula Z-R^x-boronic acid under the conditions described above for the preparation of compounds of Formula (I) and (II) from (IX) and R^γ-R^x-boronic acid. Boronic acids Z-R^x-boronic acid, R^γ-R^x-boronic acid and R^γ-boronic acid are commercially available or may be prepared by analogy to methods provided in Organometallics (1983) 2, 1316, Chem Revs. (1995) 95, 2457, Journal of Org Chem (2004) 69, 1999, SynLett (2004) (5), 892, Bioorg Med Chem (2005) 13, 2305, Tetrahedron Letters (2004) 44, 9359 and Tetrahedron Letters (2005) 45, 6657.

Compounds of Formula (I) or (II) in which R¹ represents a 4-(furopyridine)phenyl, may be prepared by treatment of a compound of Formula (II)"

in which R¹ represents a 4-ethynylphenyl derivative, and R², R³, and A are as defined above for Formula (II), with a suitable pyridine (the pyridine being substituted with adjacent hydroxy and iodo groups), with a suitable catalyst such as bis(triphenylphosphine)palladium (II) chloride and copper (I) iodide, in a suitable solvent such as triethylamine or DMF. Suitably the temperature is in the range 50-80⁰C. For examples of furopyridine synthesis see Bioorganic and Medicinal Chemistry Letters (2002) 12, 1399, Synthesis (1986) 749.

Compounds of Formula (I) or (II) in which R¹ represents a 4-(pyrrolopyridine)phenyl, may be prepared by treatment of a compound of Formula (II)" in which R¹ represents 4- ethynylphenyl with an appropriate pyridine (the pyridine being substituted by adjacent amino and iodo groups), in the presence of a suitable catalyst such as bis(triphenylphosphine)palladium (II) chloride and copper (I) iodide, in a suitable solvent such as triethylamine. Suitably the temperature is in the range 50-80⁰C. For examples of pyrrolopyridine synthesis see Heterocycles (1986) 24, 31 , Tetrahedron (2003) 59, 1571 , Synlett (1992) 515.

Compounds of Formula (I) or (II) in which R¹ represents phenyl substituted by a 4- imidazo[1 ,2-a]pyridine-2-yl, may be prepared by analogy to methods described in Tetrahedron Letters (2001 ) 42, 3077.

Compounds of Formula (I) or (II) in which R¹ represents a 4-(pyrazolopyrimidine)phenyl, may be prepared by treating a compound of Formula (II)" in which R¹ represents 4-(phenyl)-1 H- pyrazole-5-amine with 1 ,1 ,3,3-tetramethoxypropane in a suitable solvent such as acetic acid, suitably the temperature is in the range 90-110⁰C.

Suitable methods for the preparation of compounds the above discussed R^γ derivatives may be found in the chemical literature, for example those described in Comprehensive Heterocyclic Chemistry, Edited by A.R. Katritzky and CW. Rees, Pergamon 1984, and Heterocyclic Chemistry, Edited by J.A. Joules and K. Mills, 4^th Ed, Blackwell Science.

Compounds of Formula R^Y-R^X-B(OR')₂ or R^Y-B(OR')₂ for use in the preparation of compounds of Formula (II) are available commercially or may be prepared from compounds of Formula (II) R^γ-R^x-hal or R^γ-hal by methods well known in the art.

Compounds of Formula R^γ-R^x-hal or R^γ-hal for use in the preparation of compounds of Formula (II) are available commercially or may be prepared by methods well known in the art. Some examples of heteroaryl halide preparation are given below (but are not limited to these examples).

A 2-(4-bromophenyl)imidazo[1 ,2-a]pyridine derivative may be prepared by analogy to methods described in Tetrahedron Letters (2001 ) 42, 3077.

A 4-(furopyridine)phenyl bromide, may be prepared by treatment of a 4-ethynylphenyl bromide with a suitable pyridine (the pyridine being substituted with adjacent hydroxy and iodo groups), with a suitable catalyst such as bis(triphenylphosphine)palladium (II) chloride and copper (I) iodide, in a suitable solvent such as triethylamine or DMF. Suitably the temperature is in the range 50-80⁰C. For examples of furopyridine synthesis see Bioorganic and Medicinal Chemistry Letters (2002) 12, 1399, Synthesis (1986) 749.

A 4-(pyrazolopyrimidine)phenyl bromide may be prepared by treating a 3-(4-bromophenyl)- 1 H-pyrazole-5-amine with 1 ,1 ,3,3-tetramethoxypropane in a suitable solvent such as acetic acid, suitably the temperature is in the range 90-110⁰C.

Esters of compounds of Formula (I), in which A is -OR where R is selected from straight or branched chain alkyl, aralkyl, aryloxyalkyl, or aryl, may also be prepared by esterification of a compound of Formula (I) in which A is hydroxy by standard literature procedures for esterification.

It will be appreciated that compounds of Formula (I), (II), (III), (IV), (VIII), (IX) and (X) which exist as diastereoisomers may optionally be separated by techniques well known in the art, for example by column chromatography or recrystallisation. For example, the formation of an ester using a chiral alcohol, separation of the resulting diastereoisomers, and subsequent hydrolysis of the ester to yield the individual enantiomeric acid of Formula (I) (II), (III), (IV), (VIII), (IX) and (X).

It will be appreciated that racemic compounds of Formula (I), (II), (III), (IV), (VIII), (IX) and (X) may be optionally resolved into their individual enantiomers. Such resolutions may conveniently be accomplished by standard methods known in the art. For example, a racemic compound of Formula (I), (II), (III), (IV), (VIII), (IX) and (X) may be resolved by chiral preparative HPLC. Alternatively, racemic compounds of Formula (I), (II), (III), (IV), (VIII), (IX) and (X) which contain an appropriate acidic or basic group, such as a carboxylic acid group or amine group may be resolved by standard diastereoisomeric salt formation with a chiral base or acid reagent respectively as appropriate. Such techniques are well established in the art. For example, a racemic compound may be resolved by treatment with a chiral acid such as (R)-(-)-1 ,1 '-binaphthyl-2,2'-diyl-hydrogen phosphate or (-)-di-O,O'-p-tolyl-L-tartaric acid, in a suitable solvent, for example isopropanol. The free enantiomer may then be obtained by treating the salt with a suitable base, for example triethylamine, in a suitable solvent, for example methyl te/f-butyl ether. Alternatively, racemic acid compounds may be resolved using a chiral base, for example (S)-alpha methylbenzylamine, (S)-alpha phenylethylamine, (1 S, 2S)-(+)-2-amino-1-phenyl-1 ,3-propane-diol, (-) ephidrine, quinine, brucine. Individual enantiomers of Formula (II), (III), (IV), (VIII), (IX) and/or (X) may then be progressed to an enantiomeric compound of Formula (I) by the chemistry described above in respect of racemic compounds.

With appropriate manipulation and protection of any chemical functionality, synthesis of compounds of Formula (I) is accomplished by methods analogous to those above and to those described in the Experimental section. Suitable protecting groups can be found, but are not restricted to, those found in T W Greene and P G M Wuts 'Protective Groups in Organic Synthesis', 3^rd Ed (1999), J Wiley and Sons.

Various of the synthetic procedures described above in general terms (and below in specific terms) may involve heating the reactants. It will be appreciated that heating may be carried out by various conventional methods but also with the use of a microwave reactor.

EXAMPLES

ABBREVIATIONS

AcOH acetic acid

Biotage Biotage flash equipment for use with pre-packed silica cartridges

DBU 1 ,8-Diazabicyclo[5.4.0]undec-7-ene

DCE 1 ,2-dichloroethane

DCM dichloromethane

DMAP 4-dimethylaminopyridine

DME 1 ,2-dimethoxyethane

DMF Λ/,Λ/-dimethylformamide

EtOAc ethyl acetate

Et₂O diethyl ether h hours

HCI hydrochloric acid

HPLC high pressure liquid chromatography ISCO Companion Automated flash chromatography equipment with fraction analysis by UV absorption available from Presearch.

IPA isopropyl alcohol LDA Lithium diisopropylamide MDAP HPLC reverse phase HPLC on a Ci₈ column using a two-solvent gradient elution with (A) water containing formic acid (0.1%) and (B) acetonitrile-water (95:5 v/v) containing formic acid

(0.05%) as the eluents, and analysis of the fractions by electrospray mass spectroscopy. MeCN acetonitrile

MeOH methanol mins minutes

NH₂ SPE aminopropyl ion exchange cartridge

OASIS HLB cartridge Sample extraction cartridge available from Waters PdCI₂(dppf) [1 ,1 '-Bis(diphenylphosphino)ferrocene]dichloro-palladium(ll)

ROT Rotamer

SCX-2 propylsulphonic acid ion exchange cartridge

SPE solid phase extraction column

TFA trifluoroacetic acid THF tetrahydrofuran

All mass spectroscopy was performed using electrospray as the method of ionisation.

Intermediate 1 2-(4-Bromophenyl)pyrazolo[1 ,5-a]pyrimidine

A solution of 3-(4-bromophenyl)-1 /-/-pyrazol-5-amine (5.00 g) in AcOH (80 mL) was treated with 1 ,1 ,3,3-tetramethoxypropane (4.13 g) and the mixture heated at 11 O⁰C for 1.5 h. On cooling to room temperature the precipitated solid was isolated by filtration, washed with water (3 x 10 mL) and dried in vacuo at 40⁰C. This was recrystallised from acetic acid and dried in vacuo at 40⁰C to give the title compound. MS calcd for (Ci₂H₈BrN₃+ H)⁺: 274/276 MS found (electrospray): (M+H)⁺ = 274/276

Intermediate 2

2-[4-(4,4,5,5-Tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl]pyrazolo[1,5-a]pyrimidine

A mixture of Intermediate 1 (100 mg), bis(pinacolato)diboron (134 mg), potassium acetate (108 mg) and 1 ,1 '-bis (diphenylphosphino)ferrocene dichloropalladium (II) (20 mg) in dry 1 ,4- dioxane (2 ml.) was heated to 100°C under nitrogen for 15 h. The solvent was evaporated and the residue partitioned between water (10 ml.) and DCM (20 ml_). The aqueous phase was extracted further with DCM (10 mL) and the combined organics evaporated. This was purified by SPE chromatography, eluting with cyclohexane/EtOAc (3:1 ) to give the title compound.

MS calcd for (Ci₈H₂₀BN₃O₂+ H)⁺: 322 MS found (electrospray): (M+H)⁺ = 322

Intermediate 3 2-(4-Bromophenyl)furo[3,2-b]pyridine

T )> <( ))— Br

To 4-bromophenylactetylene (4.3 g) was added 2-iodo-3-hydroxypyridine (5.25 g), bis(triphenylphosphine)palladium dichloride (1.5 g), copper (I) iodide (0.58 g) and triethylamine (100 mL). The reaction was stirred at 9O⁰C, under nitrogen, for 4 h. The reaction was cooled, diluted with EtOAc (400 mL) and washed twice with saturated ammonium chloride solution, brine and then concentrated. The crude product was dissolved in DCM, filtered and purified by ISCO Companion silica chromatography eluting with a gradient of ethyl acetate in cyclohexane (30-100%) to give the title compound. MS calcd for (Ci₃H₈BrNO + H)⁺: 274/276 MS found (electrospray): (M+H)⁺ = 274/276

Intermediate 4 2-[4-(4,4,5,5-Tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl]furo[3,2-fe]pyridine

A mixture of Intermediate 3 (250 mg), bis(pinacolato)diboron (345 mg), potassium acetate (265 mg) and 1 ,1 '-bis (diphenylphosphino)ferrocene dichloro palladium (II) (48 mg) in dry 1 ,4-dioxane (5 mL) was heated to 100°C under nitrogen for 7 h. The solvent was evaporated and partitioned between water (10 mL) and DCM (20 mL). The aqueous phase was extracted further with DCM (10 mL) and the combined organics evaporated. The residue was purified by SPE chromatography eluting with cyclohexane/EtOAc (2:1 then 1 :1 ). Further purification by SPE chromatography, eluting with cyclohexane then cyclohexane/EtOAc (3:1 ) to (1 :1 ) to (1 :2) then EtOAc, EtOAc/MeCN (1 :1 ) and finally MeCN gave the title compound. MS calcd for (Ci₉H₂₀BNO₃+ H)⁺: 322 MS found (electrospray): (M+H)⁺ = 322

Intermediate 5 2-(4-Bromophenyl)pyrazolo[1,5-a]pyrimidin-7 -amine

5-Amino-3-(4-bromophenyl)pyrazole (5 g) was dissolved in acetic acid (80 ml.) and trans-3- (dimethylamino)-acryonitrile (2.3 ml.) was added. The reaction was heated to 110⁰C for 3 h., and was then allowed to cool to room temperature before being evaporated in vacuo. The residue was treated with saturated sodium bicarbonate solution and the resulting precipitate was filtered off, washed with water and dried in a vacuum oven for 18 h to give the title compound.

MS calcd for (Ci₂H₁₉N₄Br+ H) ⁺ : 289/291 MS found (electrospray): (M+H)⁺= 289/291

Intermediate 6

2-[4-(4,4,5,5-Tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl]pyrazolo[1,5-a]pyrimidin-7- amine

Intermediate 5 (1.5 g) was dissolved in dry 1 ,4-dioxane (30 ml_). To this stirred solution was added potassium acetate (1.42 g), bis(pinacolato)diboron (1.71 g) and 1 ,1 '-bis (diphenylphosphino)ferrocene dichloro palladium (II) (254 mg). The reaction mixture was then heated to 100⁰C, and stirred under nitrogen for 18 h. The reaction was then cooled and recharged with potassium acetate (1.42 g), bis(pinacolato)diboron (1.713g) and 1 ,1 '-bis (diphenylphosphino)ferrocene dichloro palladium (II) (254 mg). The reaction mixture was then reheated to 100⁰C, and stirred under nitrogen for another 18 h. The reaction was allowed to cool and the solvent removed by evaporation and the residue was partitioned between water and DCM. The organic layer was separated, washed with water, and concentrated. The residue was then purified by ISCO Companion silica chromatography eluting with a gradient of EtOAc /cyclohexane (0% to 100%), to give the title compound. MS calcd for (Ci₈H₂₁BN₄O₂+ H)⁺: 337 MS found (electrospray): (M+H)⁺ = 337

Intermediate 7 2-[4-(4,4,5,5-Tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl]imidazo[1 ,2-a]pyridine

A mixture of 2-(4-bromophenyl)imidazo[1 ,2-a]pyridine² (300 mg), bis(pinacolato)diboron (508 mg), potassium acetate (393 mg) and 1 ,1 '-bis (diphenylphosphino)ferrocene dichloro palladium (II) (58 mg) in dry 1 ,4-dioxane (6 mL) was heated to 100⁰C under nitrogen for 24 h. The solvent was evaporated and the residue was partitioned between water (10 mL) and DCM (20 mL). The aqueous phase was extracted further with DCM (10 mL) and the combined organics evaporated. The residue was purified by SPE chromatography eluting with cyclohexane/EtOAc (3:1 ). Further purification by SPE (silica) eluting with cyclohexane then cyclohexane/EtOAc [(3:1 ) followed by (1 :1 )] gave the title compound. MS calcd for (Ci₉H₂IBN₂O₂+ H)⁺: 321 MS found (electrospray): (M+H)⁺ = 321

Ref 2: Burkholder, Conrad; Dolbier, William R.; Medebielle, Maurice; Ait-Mohand, Samia, Tetrahedron Lett, 42, 17, 2001 , 3077 - 3080.

Intermediate 8

6-[4-(4,4,5,5-Tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl]imidazo[2,1 -b][1 ,3]thiazole

6-(4-lodophenyl)imidazo[2,1-b][1 ,3]thiazole (2 g) was dissolved in dry 1 ,4-dioxane (40 mL). To this stirred solution was added potassium acetate (1.8 g), bis(pinacolato)diboron (2.34 g) and 1 ,1 '-bis (diphenylphosphino)ferrocene dichloro palladium (II) (350 mg). The reaction mixture was heated to 100⁰C, and stirred under nitrogen for 18 h. Further potassium acetate (1.8 g), bis(pinacolato) diboron (2.3 g) and 1 ,1 '-bis (diphenylphosphino)ferrocene dichloro palladium (II) (350 mg) were added. The reaction mixture was then left to stir at 100⁰C for a further 24 h. The solvent was removed by evaporation and the residue was partitioned between water and DCM. The layers were separated using a hydrophobic frit, and the organic phase was concentrated by evaporation to give a solid. This was purified by ISCO Companion silica chromatography eluting with a gradient of EtOAc / cyclohexane (10% to 50%), to give the title compound.

MS calcd for (Ci₇H₁₉BN₂O₂S+ H)⁺: 327 MS found (electrospray): (M+H)⁺ = 327

Intermediate 9 Methyl 3-{[(frans-4-methylcyclohexyl)carbonyl]amino}-2-thiophenecarboxylate

Triethylamine (12.01 g), followed by a solution of frans-4-methylcyclohexanecarbonyl chloride¹ (19 mL) was added slowly to a solution of methyl 3-amino-2-thiophenecarboxylate (12.44 g) in dry DCM (200 ml_). The solution was allowed to reach 35⁰C and then stirred under nitrogen for 16 h at room temperature. After this time, the mixture was washed with saturated sodium bicarbonate solution and extracted with DCM. The combined organics were dried (hydrophobic frit) and evaporated. This was purified by silica chromatography (50 g Si Biotage) eluting with EtOAc/cyclohexane (1 :20) to give the title compound. MS calcd for (C₂₃H₂₈CINO₃S+ H)⁺: 282 MS found (electrospray): (M+H)⁺ =282 Ref 1 : WO 2004/052885.

Intermediate 10

Methyl S-I^frans^-methylcyclohexylJcarbonylJ^etrahydro-S-furanylJamino]^- thiophenecarboxylate

3-Hydroxytetrahydrofuran (7.84 ml.) was dissolved in dry DCM (400 ml.) under nitrogen and pyridine (10.2 ml.) was added. The reaction mixture was then cooled to -1O⁰C. Trifluoromethylsulphonyl anhydride (19.6 ml.) was added dropwise and the mixture stirred for 1 h (solution A).

To a solution of methyl 3-{[(frans-4-methylcyclohexyl)carbonyl]amino}-2- thiophenecarboxylate (a synthesis of which is described above as Intermediate 9) (3 g) in dry THF (60 ml.) at -78⁰C under nitrogen was added N,N-dimethylformamide (7.5 ml_), followed by KHMDS solution (28 ml_, 0.5 M. in toluene) dropwise. The reaction mixture was stirred under nitrogen for 0.5 h at -78⁰C, and then allowed to reach -1 O⁰C and maintained at that temperature for 1.5 h. Solution A (22 ml.) was added dropwise at -1O⁰C and the reaction mixture was stirred for 0.5 h, and then allowed to reach room temperature. This was then quenched with saturated sodium bicarbonate solution. The organic phase was separated, washed with brine, dried over sodium sulphate, filtered and evaporated. The residue was purified by ISCO Companion silica chromatography, eluting with a gradient of EtOAc in cyclohexane (5% to 100%) to give the title compound. MS calcd for (Ci₇H₂₄NIO₃S + H)⁺: 352 MS found (electrospray): (M+H)⁺ = 352

Intermediate 11 Method A

Methyl S-iodo-S-IKfrans^-methylcyclohexylJcarbonylKtetrahydro-S-furanylJamino]^- thiophenecarboxylate

A solution of methyl 3-[[(frans-4-methylcyclohexyl)carbonyl](tetrahydro-3-furanyl)amino]-2- thiophenecarboxylate (a synthesis of which is described above as Intermediate 10) (263 mg) in dry THF (3.2 ml.) was added dropwise at -78°C under nitrogen to a 2M solution of LDA in THF/heptane/ethyl benzene (0.75 ml.) maintaining an internal temperature of -78°C. The reaction was stirred at -78°C for 2 h. A solution of iodine (380 mg) in dry THF (3.2 ml.) was added dropwise to the stirred reaction mixture maintaining an internal temperature of -78°C. After stirring under nitrogen at -78°C for 1 h, the reaction mixture was quenched by addition of saturated ammonium chloride solution (1 ml.) and warmed to 0⁰C. The mixture was diluted with 5% sodium thiosulfate solution, the organic phase was separated and the aqueous phase was extracted with EtOAc. The combined organic phases were dried (Na₂SO₄), filtered and evaporated. The residue was purified by ISCO Companion silica chromatography, eluting with a gradient of EtOAc in cyclohexane (5% to 100%) to give the title compound. MS calcd for (Ci₇H₂₄NIO₃S + H)⁺: 478 MS found (electrospray): (M+H)⁺ = 478

Alternative Preparation of Intermediate 11 Method B Methyl S-iodo-S-IKfrans^-methylcyclohexylJcarbonylKtetrahydro-S-furanylJamino]^- thiophenecarboxylate

Intermediate 40 (78 mg) was dissolved in DCM (2 ml.) and trans-4- methylcyclohexanecarbonyl chloride¹ (43 mg) was added. The reaction was stirred and heated at 85⁰C under nitrogen for 20 h. The reaction was allowed to cool to room temperature and was quenched with saturated sodium bicarbonate solution. The organics were extracted with DCM, separated using a hydrophobic frit and evaporated in vacuo. The crude material was purified using ISCO Companion silica chromatography, eluting with a gradient 5-100% EtOAc in cyclohexane to give the title compound. MS calcd for (Ci₈H₂₄INO₄S + H)⁺: 478 MS found (electrospray): (M+H)⁺ = 478 Ref 1 : WO 2004/052885.

Intermediate 12 Methyl 3-(tetrahydro-2H-pyran-4-ylamino)-2-thiophenecarboxylate

Tetrahydro-4H-pyran-4-one (5.87 mL) was added to a solution of methyl-3-amino-2 thiophene carboxylate (5 g) in dry DCM (100 mL) at room temperature under nitrogen.

Glacial acetic acid (5.46 mL) was added slowly. Sodium triacetoxyborohydride (10.11 g) was then added in portions over 30 min. The resulting solution was left to stir at room temperature for 20 h. The mixture was treated with 8% sodium bicarbonate solution slowly

(300 mL), the layers were separated and the DCM layer washed further with sodium bicarbonate solution (2 x 100 mL), dried (hydrophobic frit) and evaporated to give the crude product. This was purified by ISCO Companion silica chromatography eluting with a gradient of EtOAc /cyclohexane (5% to 80%), to give the title compound.

MS calcd for (C_H H₁₅SNO₃+ H)⁺: 242

MS found (electrospray): (M+H)⁺ = 242

Intermediate 13

Methyl S-IKfrans^-methylcyclohexylJcarbonylKtetrahydro^H-pyran^-ylJamino]^- thiophenecarboxylate

Triphenylphosphine (5.52 g) was added in portions to a solution of intermediate 12 (2.99 g) in dry DCM (48 mL) at room temperature under nitrogen. trans-4-

Methylcyclohexanecarbonyl chloride¹ (3.98 mL) was added in 1 mL portions. The solution was heated to 45⁰C under nitrogen for 24 h. Further frans-4-methylcyclohexanecarbonyl chloride¹ (1 mL) was added and heating continued for 16 h. The mixture was cooled, treated with saturated sodium bicarbonate solution and stirred at room temperature to neutralise for 1 h. The layers were separated and the organic layer was washed with saturated bicarbonate solution, dried (hydrophobic frit) and evaporated. This was purified by ISCO Companion silica chromatography eluting with a gradient of EtOAc /cyclohexane (10% to 70%), to give the title compound. MS calcd for (Ci₉H₂₇SNO₄+ H)⁺: 366 MS found (electrospray): (M+H)⁺ = 366

Intermediate 14

Methyl S-iodo-S-IKfrans^-methylcyclohexylJcarbonylKtetrahydro^H-pyran^- yl)amino]-2-thiophenecarboxylate

A solution of Intermediate 13 (2.87 g) in dry THF (29 ml.) was added dropwise at -78°C under nitrogen to a 2M solution of LDA in THF/heptane/ethyl benzene (7.85 mL) maintaining an internal temperature -78°C. The reaction was stirred at -78°C for 2 h. A solution of iodine (3.99 g) in dry THF (29 mL) was added dropwise to the stirred reaction mixture maintaining an internal temperature -78°C. After stirring under nitrogen at -78°C for 0.25 h, the reaction mixture was quenched by addition of saturated ammonium chloride solution (1 mL) and warmed to 0⁰C. The mixture was diluted with 5% sodium thiosulfate solution, the organic phase was separated and the aqueous phase was extracted with EtOAc. The combined organic phases were dried (Na₂SO₄), filtered and evaporated. The residue was purified by ISCO Companion silica chromatography, eluting with a gradient of EtOAc in cyclohexane (20% to 80%) to give the title compound. MS calcd for (Ci₇H₂₄NIO₃S + H)⁺: 491.8 MS found (electrospray): (M+H)⁺ = 491.8

Intermediate 15

Methyl S-IKfrans^-methylcyclohexylJcarbonylKtetrahydro-S-furanylJaminol-S-^- pyrazolo[1,5-a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylate

2-[4-(4,4,5,5-Tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl]pyrazolo[1 ,5-a]pyrimidine (a synthesis of which is described above as Intermediate 2) (71 mg), methyl 5-iodo-3-[[(frans-4- methylcyclohexyl)carbonyl](tetrahydro-3-furanyl)amino]-2-thiophenecarboxylate (a synthesis of which is described above as Intermediate 11 ) (75 mg), sodium carbonate (67 mg) and tetrakis(triphenylphosphine) palladium (18 mg) in DMF (6.75 mL) were heated at 100°C under a nitrogen atmosphere for 3 h. The solvent was evaporated and the mixture was partitioned between water (10 mL) and DCM (15 mL). The combined organic phases was passed through a hydrophobic frit and evaporated. This was purified by ISCO Companion silica chromatography eluting with a gradient of EtOAc /cyclohexane (20% to 100%), to give the title compound. MS calcd for (C₃₀H₃₂SN₄O₄+ H)⁺: 545 MS found (electrospray): (M+H)⁺ = 545

Intermediate 16 Methyl S^^frans^-methylcyclohexylJcarbonylJ^^methyloxyJethylJamino}^- thiophenecarboxylate

A solution of methyl3-{[(frans-4-methylcyclohexyl)carbonyl]amino}-2-thiophenecarboxylate (a synthesis of which is described above as Intermediate 9) (12.0 g) in dry DMF (350 ml. ) was stirred at room temperature under nitrogen and treated portion-wise with sodium hydride (2.56 g, 60% in oil). The reaction mixture was stirred for 10 mins then treated with a solution of bromoethylmethyl ether (6.4 ml.) in dry DMF (10 ml_). The reaction was stirred at 6O⁰C for 16 h. The reaction was allowed to cool and concentrated in vacuo. The residue was partitioned between EtOAc and water and the aqueous was rextracted with EtOAc and the combined organics dried over sodium sulphate, and evaporated. This was then purified by ISCO Companion silica chromatography eluting with a gradient of EtOAc /cyclohexane (0% to 70%), to give the title compound MS calcd for (Ci₇H₂₅SNO₄+ H)⁺: 340 MS found (electrospray): (M+H)⁺ = 340

Intermediate 17

Methyl S-iodo-S-fl^rans^-methylcyclohexylJcarbonyl^-fmethyloxyJethyllamino}^- thiophenecarboxylate

A solution of Intermediate 16 (3.7 g in d (40 ml.) was added dropwise at -70⁰C under nitrogen to a 2M solution of LDA in THF/heptane/ethyl benzene (16.4 ml.) maintaining an internal temperature -78°C. The reaction was stirred at -70⁰C for 2 h. A solution of iodine (5.56 g) in dry THF (40 ml.) was added dropwise to the stirred reaction mixture maintaining an internal temperature -70⁰C. After stirring under nitrogen at -78°C for 1 h, the reaction mixture was quenched by addition of saturated ammonium chloride solution (75 ml.) and warmed to 0°C. The mixture was diluted with 5% sodium thiosulfate solution (75 ml_), then the organic phase was separated and the aqueous phase was extracted with EtOAc. The combined organic phases were dried (Na₂SO₄), filtered and evaporated. The residue was purified by ISCO Companion silica chromatography, eluting with a gradient of EtOAc in cyclohexane (0% to 30%) to give the title compound. MS calcd for (Ci₇H₂₄NIO₄S + H)⁺: 466 MS found (electrospray): (M+H)⁺ = 466 Intermediate 18

Methyl S-^-furoIS^-felpyridin^-ylphenylJ-S-fl^rans^-methylcyclohexylJcarbonyl]^-

(methyloxy)ethyl]amino}-2-thiophenecarboxylate

2-[4-(4,4,5,5-Tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl]furo[3,2-ιb]pyridine (a synthesis of which is described above as Intermediate 4) (0.36 g), methyl 5-iodo-3-{[(frans-4- methylcyclohexyl)carbonyl][2-(methyloxy)ethyl]amino}-2-thiophenecarboxylate (a synthesis of which is described above as Intermediate 17) (0.4 g), 2N sodium carbonate (2 ml.) and tetrakis(triphenylphosphine) palladium (139 mg) in DMF (6 ml.) were heated at 100°C under a nitrogen atmosphere for 2 h. The reaction was allowed to cool, the solvent removed and the mixture partitioned between water and EtOAc. The aqueous re-extracted with more EtOAc and the combined organic phases were washed with water, brine and dried over sodium sulphate, and evaporated. This was then purified by ISCO Companion silica chromatography eluting with a gradient of EtOAc /cyclohexane (0% to 100%), to give the title compound.

MS calcd for (C₃₀H₃₂SN₂O₅+ H)⁺: 533 MS found (electrospray): (M+H)⁺ = 533

Intermediate 19 Methyl 5-[4-(7-aminopyrazolo[1 ,5-a]pyrimidin-2-yl)phenyl]-3-{[(frans-4- methylcyclohexyl)carbonyl][2-(methyloxy)ethyl]amino}-2-thiophenecarboxylate

2-[4-(4,4,5,5-Tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl]pyrazolo[1 ,5-a]pyrimidin-7-amine (a synthesis of which is described above as Intermediate 6) (0.37 g), methyl 5-iodo-3-{[(frans-4- methylcyclohexyl)carbonyl][2-(methyloxy)ethyl]amino}-2-thiophenecarboxylate (a synthesis of which is described above as Intermediate 17) (0.4 g), 2N sodium carbonate solution (2 ml.) and tetrakis(triphenylphosphine) palladium (139 mg) in DMF (6 ml.) were heated at 100°C under a nitrogen atmosphere for 2 h. The reaction was allowed to cool, the solvent removed and the mixture was partitioned between water and EtOAc. The aqueous re- extracted with more EtOAc and the combined organic phases were washed with water, brine and dried over sodium sulphate, and evaporated. This was then purified by ISCO Companion silica chromatography eluting with a gradient of EtOAc /cyclohexane (0% to 20%), to give the title compound. MS calcd for (C₂₉H₃₃SN₅O₄+ H)⁺: 548 MS found (electrospray): (M+H)⁺ = 548

Intermediate 20

Methyl 3-{[(frans-4-methylcyclohexyl)carbonyl][2-(methyloxy)ethyl]amino}-5-(4- pyrazolo[1,5-a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylate

2-[4-(4,4,5,5-Tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl]pyrazolo[1 ,5-a]pyrimidine (a synthesis of which is described above as Intermediate 2) (0.34g), Intermediate 17 (0.4 g), 2N sodium carbonate solution (2 ml) and tetrakis(triphenylphosphine) palladium (139 mg) in DMF (6 ml.) were heated at 100°C under a nitrogen atmosphere for 2 h. The reaction was allowed to cool, the solvent removed and the mixture was partitioned between water and EtOAc. The aqueous re-extracted with more EtOAc and the combined organic phases were washed with water, brine and dried over sodium sulphate, and evaporated. This was then purified by ISCO Companion silica chromatography eluting with a gradient of EtOAc /cyclohexane (0% to 100%), to give the title compound. MS calcd for (C₂₉H₃₂SN₄O₄+ H)⁺: 533 MS found (electrospray): (M+H)⁺ = 533

Intermediate 21

Methyl 5-(4-imidazo[1 ,2-a]pyridin-2-ylphenyl)-3-{[(frans-4- methylcyclohexyl)carbonyl][2-(methyloxy)ethyl]amino}-2-thiophenecarboxylate

2-[4-(4,4,5,5-Tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl]imidazo[1 ,2-a]pyridine (a synthesis of which is described above as Intermediate 7) (0.44 g), Intermediate 17 (0.53 g), 2N sodium carbonate solution (2.65 ml.) and tetrakis(triphenylphosphine) palladium (186 mg) in DMF (10 ml.) were heated at 100°C under a nitrogen atmosphere for 1.5 h. The reaction was allowed to cool, solvent removed and the mixture was partitioned between water and EtOAc . The aqueous re-extracted with more EtOAc and the combined organic phases were washed with water, brine and dried over sodium sulphate, and evaporated. This was purified by ISCO Companion silica chromatography eluting with a gradient of EtOAc /cyclohexane (0% to 100%), to give the title compound. MS calcd for (C₃₀H₃₃SN₃O₄+ H)⁺: 532 MS found (electrospray): (M+H)⁺ = 532

Intermediate 22

Methyl 5-(4-imidazo[2,1 -fe][1, 3]thiazol-6-ylphenyl)-3 -{[(frans-4- methylcyclohexyl)carbonyl][2-(methyloxy)ethyl]amino}-2-thiophenecarboxylate

6-[4-(4,4,5,5-Tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl]imidazo[2,1-b][1 ,3]thiazole (a synthesis of which is described above as Intermediate 8) (0.46 g), Intermediate 17 (0.5 g), 2N sodium carbonate solution (2.5 ml.) in water and tetrakis(triphenylphosphine) palladium (173 mg) in DMF (6 ml.) were heated at 100°C under a nitrogen atmosphere for 2 h. The reaction was allowed to cool, the solvent removed and the mixture was partitioned between water and EtOAc. The aqueous was re-extracted with more EtOAc and the combined organic phases were washed with water, brine and dried over sodium sulphate, and evaporated. This was purified by ISCO Companion silica chromatography eluting with a gradient of EtOAc /cyclohexane (0% to 100%), to give the title compound. MS calcd for (C₂₈H₃₁S₂N₃O₄+ H)⁺: 538 MS found (electrospray): (M+H)⁺ = 538

Intermediate 23

Methyl S^^^ethyloxyJethylJ^frans^-methylcyclohexylJcarbonylJamino}^- thiophenecarboxylate

A solution of Intermediate 9 (6.0 g) in dry DMF( 200 ml. ) was stirred at O⁰C under nitrogen and treated portionwise with sodium hydride (0.94 g, 60% in oil). The reaction mixture was stirred at room temperature for 1 h then treated with a solution of bromoethylethyl ether (2.67 ml_) in dry DMF (10 ml_). The reaction was stirred at 9O⁰C for 24 h and 21⁰C for another 24 h. The reaction was quenched with methanol and concentrated in vacuo. The residue was partitioned between EtOAc and water and the aqueous was re-extracted with EtOAc and the combined organics dried over sodium sulphate, and evaporated. This was purified by ISCO Companion silica chromatography eluting with a gradient of EtOAc /cyclohexane (0% to 60%), to give the title compound. MS calcd for (Ci₈H₂₇SNO₄+ H)⁺: 354 MS found (electrospray): (M+H)⁺ = 354

Intermediate 24

Methyl 3-{[2-(ethyloxy)ethyl][(frans-4-methylcyclohexyl)carbonyl]amino}-5-iodo-2- thiophenecarboxylate

n-Butyl lithium (10 ml_, 1.6 mol in hexanes) was added dropwise to diisiopropylamine (2.27 ml.) at O⁰C under nitrogen, followed by the addition of 10 ml. dry THF. The reaction mixture was stirred at O⁰C for 10 mins and then cooled to -78⁰C. A solution of Intermediate 23 (2.84 g) in dry THF (30 ml.) was added dropwise under nitrogen to the solution of the LDA maintaining an internal temperature -78°C. The reaction was stirred at -78°C for 1 h. A solution of iodine (3.67g) in dry THF (30 ml.) was added dropwise to the stirred reaction mixture maintaining an internal temperature of -78°C. After stirring under nitrogen at -78°C for 1 h, the reaction mixture was quenched by addition of saturated ammonium chloride solution (55 ml.) and warmed to -5°C. The mixture was diluted with 5% sodium thiosulfate solution (75 ml_), the organic phase was separated and the aqueous phase was extracted with EtOAc. The combined organic phases were washed with brine, dried (Na₂SO₄), filtered and evaporated. The residue was purified by ISCO Companion silica chromatography, eluting with a gradient of EtOAc in cyclohexane (0% to 60%) to give the title compound. MS calcd for (Ci₈H₂₆NIO₄S + H)⁺: 480 MS found (electrospray): (M+H)⁺ = 480

Intermediate 25

Methyl 3-{[2-(ethyloxy)ethyl][(frans-4-methylcyclohexyl)carbonyl]amino}-5-(4-furo[3,2- fe]pyridin-2-ylphenyl)-2-thiophenecarboxylate

2-[4-(4,4,5,5-Tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl]furo[3,2-b]pyridine (a synthesis of which is described above as Intermediate 4) (0.48 g), Intermediate 24 (0.55 g), 2N sodium carbonate (2.65 ml.) and tetrakis(triphenylphosphine) palladium (186 mg) in DMF (6 ml.) were heated at 100°C under a nitrogen atmosphere for 2 h. The reaction was allowed to cool, solvent removed and the mixture partitioned between water and EtOAc. The aqueous re-extracted with more EtOAc and the combined organic phases were washed with water, brine and dried over sodium sulphate, and evaporated. This was purified by ISCO Companion silica chromatography eluting with a gradient of EtOAc /cyclohexane (0% to 100%), to give the title compound. MS calcd for (C₃IH₃₄SN₂O₅+ H)⁺: 547 MS found (electrospray): (M+H)⁺ = 547

Intermediate 26

Methyl 5-[4-(7-aminopyrazolo[1,5-a]pyrimidin-2-yl)phenyl]-3-{[2-(ethyloxy)ethyl][(frans- 4-methylcyclohexyl)carbonyl]amino}-2-thiophenecarboxylate

2-[4-(4,4,5,5-Tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl]pyrazolo[1 ,5-a]pyrimidin-7-amine (a synthesis of which is described above as Intermediate 6) (0.5 g), Intermediate 24 (0.55 g), 2N sodium carbonate solution (2 ml.) and tetrakis(triphenylphosphine) palladium (186 mg) in DMF (2.65 ml.) were heated at 100°C under a nitrogen atmosphere for 2.5 h. The reaction was allowed to cool, solvent removed and the mixture was partitioned between water and EtOAc. The aqueous re-extracted with more EtOAc and the combined organic phases were washed with water, brine and dried over sodium sulphate, and evaporated. This was purified by ISCO Companion silica chromatography eluting with a gradient of EtOAc /cyclohexane (0% to 100%), to give the title compound. MS calcd for (C₃₀H₃₅SN₅O₄+ H)⁺: 562 MS found (electrospray): (M+H)⁺ = 562

Intermediate 27 Methyl 3-{[2-(ethyloxy)ethyl][(frans-4-methylcyclohexyl)carbonyl]amino}-5-(4- pyrazolo[1,5-a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylate

2-[4-(4,4,5,5-Tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl]pyrazolo[1 ,5-a]pyrimidine (a synthesis of which is described above as Intermediate 2) (0.48 g), methyl 3-{[2-(ethyloxy) ethyl][(frans-4-methylcyclohexyl)carbonyl]amino}-5-iodo-2-thiophenecarboxylate (a synthesis of which is described above as Intermediate 24 (0.55 g), 2N sodium carbonate solution (2.65 ml.) and tetrakis(triphenylphosphine) palladium (186 mg) in DMF (10 ml.) were heated at 100⁰C under a nitrogen atmosphere for 1.5 h. The reaction was allowed to cool, the solvent removed and the mixture was partitioned between water and EtOAc. The aqueous was re- extracted with more EtOAc and the combined organic phases were washed with water, brine and dried over sodium sulphate, and evaporated. This was purified by ISCO Companion silica chromatography eluting with a gradient of EtOAc /cyclohexane (0% to 100%), to give the title compound. MS calcd for (C₃₀H₃₄SN₄O₄+ H)⁺: 547 MS found (electrospray): (M+H)⁺ = 547

Intermediate 28

Methyl 3-{[2-(ethyloxy)ethyl][(frans-4-methylcyclohexyl)carbonyl]amino}-5-(4- imidazo[1,2-a]pyridin-2-ylphenyl)-2-thiophenecarboxylate

Intermediate 7 (0.44 g), Intermediate 24 (0.55 g), 2N sodium carbonate solution (2.65 ml.) and tetrakis(triphenylphosphine) palladium (186 mg) in DMF (10 ml.) were heated at 100°C under a nitrogen atmosphere for 1.5 h. The reaction was allowed to cool, solvent removed and the mixture was partitioned between water and EtOAc. The aqueous was re-extracted with more EtOAc and the combined organic phases were washed with water, brine and dried over sodium sulphate, and evaporated. This was purified by ISCO Companion silica chromatography eluting with a gradient of EtOAc /cyclohexane (0% to 100%), to give the title compound.

MS calcd for (C₃i H₃₅SN₃O₄+ H)⁺: 546 MS found (electrospray): (M+H)⁺ = 546

Intermediate 29

Methyl S-^-fethyloxyJethyllKfrans^-methylcyclohexylJcarbonyllamino}^-^- imidazo[2,1 -ϋ][1,3]thiazol-6-ylphenyl)-2-thiophenecarboxylate

Intermediate 8 (0.49 g), Intermediate 24 (0.55 g), 2N sodium carbonate solution (2.5 ml.) and tetrakis(triphenylphosphine) palladium(O) (186 mg) in DMF (10 ml.) were heated at 100°C under a nitrogen atmosphere for 2 h. The reaction was allowed to cool, solvent removed and the mixture was partitioned between water and EtOAc. The aqueous was re-extracted with more EtOAc and the combined organic phases were washed with water, brine and dried over sodium sulphate, and evaporated. This was purified by ISCO Companion silica chromatography eluting with a gradient of EtOAc /cyclohexane (0% to 100%), to give the title compound.

MS calcd for (C₂₉H₃₃S₂N₃O₄+ H)⁺: 552 MS found (electrospray): (M+H)⁺ = 552

Intermediate 30 Methyl S-I^frans^-methylcyclohexylJcarbonylJ^etrahydro^H-pyran^-ylJaminol-S- phenyl-2-thiophenecarboxylate

To a stirred solution of Intermediate 14 (300 mg) in DME (3 ml.) and water (0.75 ml_), was added phenylboronic acid (112 mg), potassium phosphate (259 mg), and tetrakis(triphenylphosphine)palladium(0) (71 mg). The reaction mixture was then heated at 8O⁰C for 1.5 h. After cooling to room temperature, the reaction mixture was partitioned between saturated sodium bicarbonate solution and ethyl acetate. The layers were separated, and the organic phase was dried over magnesium sulphate, filtered, and concentrated by evaporation. The residue was then purified by ISCO Companion silica chromatography, eluting with a gradient of ethyl acetate in cyclohexane to give the title compound.

MS calcd for (C₂₅H₃₁NO₃S + H)⁺: 442 MS found (electrospray): (M+H)⁺ = 442

Intermediate 31

Methyl 3-{[1 -methyl-2-(methyloxy)ethyl]amino}-5-phenyl-2-thiophenecarboxylate

To a solution of methyl S-amino-δ-phenyl^-thiophenecarboxylate (2.0 g; 8.58 mmol) and glacial acetic acid (5 ml.) in dry dichloromethane (100 ml.) was added methoxyacetone (0.91 g, 10.3 mmol) and the reaction stirred at room temperature under nitrogen for 10 minutes. Sodium triacetoxyborohydride (3.6 g, 17.2 mmol) was added and the reaction heated at reflux under nitrogen for 3 days. A further addition of sodium triacetoxyborohydride (1.8 g, 8.6 mmol) was added and the reaction heated at reflux under nitrogen for 2 days. After cooling to room temperature sodium bicarbonate solution (100 ml.) was added and the reaction stirred for 30 minutes, dried using a hydrophobic frit and evaporated to dryness. The crude product was purified by 120 g Si ISCO chromatography eluted EtOAc in cyclohexane (gradient from 0-100%) to give title compound. MS calcd for (Ci₆H₁₉NO₃S + H)⁺: 306 MS found (electrospray): (M+H)⁺ = 306

Intermediate 32

Methyl S-fl^rans^-methylcyclohexylJcarbonylHI -methyl^-fmethyloxyJethyllamino}^- phenyl-2-thiophenecarboxylate

To a solution of Intermediate 31 (0.17 g; 0.56 mmol) and triphenylphosphine (0.22 g; 0.84 mmol) in dry DCM (25 ml.) was added frans-4-methylcyclohexanecarbonyl chloride¹ (135 mg; 0.84 mmol). The reaction was heated to reflux under nitrogen for 4 days. A further addition of frans-4-methylcyclohexanecarbonyl chloride (45 mg; 0.28 mmol) was added and the reaction heated at reflux under nitrogen for 8 days. The reaction was cooled to room temperature, acidified with 2M HCI, taken into DCM, washed with water, dried using a hydrophobic frit and evaporated to dryness. The crude product was purified by 12 g silica ISCO chromatography eluted with 0 -100% EtOAc / cyclohexane to give the title compound. MS calcd for (C₂₄H_3INO₄S + H)⁺: 430 MS found (electrospray): (M+H)⁺ = 430

Intermediate 33

Methyl 3-({2-(methyloxy)-1 -[(methyloxy)methyl]ethyl}amino)-2-thiophenecarboxylate

1 ,3-Dimethoxyacetone (3 ml.) and glacial acetic acid (2.2 ml.) were added to a stirred solution of methyl 3-amino-2-thiophenecarboxylate (2 g) in dry DCM (40 ml_). Sodium triacetoxyborohydride (4.05 g) was then added portion-wise, and the reaction was allowed to stir at room temperature under nitrogen for 48 h. The mixture was quenched with saturated sodium bicarbonate solution, and the organics were extracted with DCM. The organics were separated using a hydrophobic frit, and were evaporated in vacuo. The crude material was purified by ISCO Companion silica chromatography, eluting with a gradient 0-60% EtOAc in cyclohexane to give the title compound. MS calcd for (CnH₁₇NO₄S + H)⁺: 260 MS found (electrospray): (M+H)⁺ = 260

Intermediate 34

Methyl 3-([(frans-4-methylcyclohexyl)carbonyl]{2-(methyloxy)-1 -

[(methyloxy)methyl]ethyl}amino)-2-thiophenecarboxylate

To a solution of methyl 3-({2-(methyloxy)-1-[(methyloxy)methyl]ethyl}amino)-2- thiophenecarboxylate (1.47 g, a synthesis of which is described above as Intermediate 33) in DCM (29.48 ml.) was added frans-4-methylcyclohexanecarbonyl chloride¹ (1.8 g) and the mixture was allowed to stir under nitrogen at 85⁰C for 18 h. A further portion of trans-4- methylcyclohexanecarbonyl chloride (0.9 ml.) was added and the mixture was allowed to stir under nitrogen at 85⁰C for 64 h. The mixture was then allowed to cool to room temperature and was quenched with saturated sodium bicarbonate solution. The organics were extracted into DCM and were separated using a hydrophobic frit. The organics were evaporated in vacuo and the residue was purified by ISCO Companion silica chromatography, eluting with a gradient 5-90% EtOAc in cyclohexane to give the title compound. MS calcd for (Ci₉H₂₉NO₅S + H)⁺: 384 MS found (electrospray): (M+H)⁺ = 384 Ref 1 : WO2004/052885

Intermediate 35 Methyl 5-iodo-3-([(frans-4-methylcyclohexyl)carbonyl]{2-(methyloxy)-1 - [(methyloxy)methyl]ethyl}amino)-2-thiophenecarboxylate

A solution of LDA (2M solution in THF/heptane/ethyl benzene, 3.13 ml.) was cooled to -78⁰C under nitrogen. A solution of Intermediate 34 (1.2 g) in dry THF (12 ml.) was added dropwise, and the dropping funnel was rinsed through with dry THF (2 ml_). The mixture was stirred at -78⁰C for 30 mins, then a solution of iodine (1.59 g) in dry THF (12 ml.) was added dropwise. The reaction was stirred at -78⁰C under nitrogen for 10 mins, and was quenched with saturated ammonium chloride solution. The mixture was allowed to warm to room temperature before being washed with 5% sodium thiosulphate solution. The organics were extracted with EtOAc and the layers were separated. The organics were washed with brine, dried over sodium sulphate, filtered through a hydrophobic frit and evaporated in vacuo. The crude material was purified by ISCO Companion silica chromatography, eluting with a gradient 5-80% EtOAc in cyclohexane to give the title compound. MS calcd for (Ci₉H₂₈INO₅S + H)⁺: 510 MS found (electrospray): (M+H)⁺ = 510

Intermediate 36

Methyl 3-([(frans-4-methylcyclohexyl)carbonyl]{2-(methyloxy)-1 - [(methyloxy)methyl]ethyl}amino)-5-(4-pyrazolo[1,5-a]pyrimidin-2-ylphenyl)-2- thiophenecarboxylate

To a stirred solution of Intermediate 35 (215 mg) in DME (2.15 ml.) and water (0.54 ml.) was added 2-[4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl]pyrazolo[1 ,5-a]pyrimidine (203 mg, a synthesis of which is described above as Intermediate 2), potassium phosphate (179 mg) and tetrakis(triphenylphosphine)palladium (0) (49 mg). The reaction mixture was stirred at 8O⁰C under nitrogen for 3.5 h. The mixture was allowed to cool to room temperature and was partitioned between saturated sodium bicarbonate solution and EtOAc. The organics were separated, washed with brine, dried using a hydrophobic frit and evaporated in vacuo. The crude material was purified by ISCO Companion silica chromatography, eluting with a gradient 20-100% EtOAc in cyclohexane to give the title compound.

MS calcd for (C_3IH₃₆N₄O₅S + H)⁺: 577 MS found (electrospray): (M+H)⁺ = 577

Intermediate 37

Methyl 3-({[(1,1 -dimethylethyl)oxy]carbonyl}amino)-2-thiophenecarboxylate

To a solution of methyl 3-amino-2-thiophenecarboxylate (5 g) in dry MeCN (100 mL) was added DMAP (389 mg). A solution of di-te/f-butyl dicarbonate (38 mL, 1 M in THF) was added slowly and the resulting solution was left to stir at room temperature under nitrogen for 20 h. 2-Amino-5-diethylaminopentane (1.54 mL) was added and the reaction mixture was stirred for 30 mins. The solvent was evaporated in vacuo and the residue was dissolved in

DCM and was washed with 1 N HCI (100 mL) and water. The organics were separated, dried over sodium sulphate and evaporated in vacuo. The crude material was purified by ISCO

Companion silica chromatography, eluting with a gradient 0-100% EtOAc in cyclohexane to give the title compound.

¹H NMR (CDCI₃) δ 9.36 (1 H, br s), 7.89 (1 H, d), 7.44 (1 H, d), 3.88 (3H, s), 1.53 (9H, s).

Intermediate 38

Methyl 3-({[(1 ,1 -dimethylethyl)oxy]carbonyl}amino)-5-iodo-2-thiophenecarboxylate

A solution of LDA (2M solution in THF/heptane/ethyl benzene, 18.96 mL) was cooled to -

78⁰C under nitrogen. A solution of Intermediate 37 (3.252 g) in dry THF (32 mL) was added slowly via dropping funnel. The mixture was stirred at -78⁰C for 10 mins, then a solution of iodine (6.4g) in dry THF (32 mL) was added dropwise. The reaction was stirred at -78⁰C under nitrogen for 1.5 h, and was quenched with saturated ammonium chloride solution. The mixture was washed with 5% sodium thiosulphate solution and the organics were extracted with EtOAc and the layers were separated. The organics were washed with brine, separated, dried over sodium sulphate and evaporated in vacuo. The crude material was purified by ISCO Companion silica chromatography, eluting with a gradient 0-60% EtOAc in cyclohexane to give the title compound.

¹H NMR (CDCI₃) δ 9.32 (1 H, br), 8.14 (1 H, s), 3.86 (3H, s), 1.52 (9H, s).

Intermediate 39

Methyl 3-amino-5-iodo-2-thiophenecarboxylate

Intermediate 38 (4.097 g) was dissolved in 4M HCI in 1 ,4-dioxane solution (40 ml_). The resulting solution was stirred under nitrogen at room temperature for 7 h. The solvent was evaporated in vacuo and the residue was partitioned between saturated sodium bicarbonate solution and DCM. The organics were separated using a hydrophobic frit and were evaporated in vacuo to give the title compound. 1H NMR (CDCI₃) δ 6.73 (1 H, s), 5.46 (2H, br), 3.82 (3H, s).

Intermediate 40 Methyl 5-iodo-3-(tetrahydro-3-furanylamino)-2-thiophenecarboxylate

A solution of Intermediate 39 (3.06 g) in dry DCM (61 ml.) was added to cyclopentanone (1.85 g). To the resulting solution was added glacial acetic acid (1.85 ml_). Sodium triacetoxyborohydride (4.57 g) was added portion-wise, and the resulting solution was stirred under nitrogen at room temperature for 20 h, then at 4O⁰C for 6 days. The reaction was allowed to cool to room temperature and was quenched with saturated sodium bicarbonate solution. The organic phase was diluted with DCM and the layers were separated. The organics were dried using a hydrophobic frit and were evaporated in vacuo. The crude material was purified by ISCO Companion silica chromatography, eluting with a gradient 5- 100% EtOAc in cyclohexane. The material was purified further by reverse phase ISCO Companion chromatography, using a C18 cartridge, eluting with a gradient 30% MeCN (0.05% formic acid)/water (0.1 % formic acid) to 80% MeCN (0.05% formic acid) to give the title compound. MS calcd for (C_I0H₁₂I NO₃S + H)⁺: 354 MS found (electrospray): (M+H)⁺ = 354

Intermediate 40a

Methyl S-iodo-S-IKfrans^-methylcyclohexylJcarbonyl^tetrahydro-S-furanylJamino]^- thiophenecarboxylate

To a solution of Intermediate 40 (78 mg) in DCM (2 mL) was added trans-4- methylcyclohexanecarbonyl chloride¹ (43 mg) and the resulting solution was heated at 85°C under nitrogen for 20 hours. The reaction was allowed to cool to room temperature and was quenched with saturated sodium bicarbonate solution. The organic phase was extracted with DCM and the layers were separated using a hydrophobic frit. The organic fraction was evaporated in vacuo and the crude material was purified by ISCO Companion silica chromatography, eluting with a gradient 5-100% EtOAc in cyclohexane to give the title compound. MS calcd for (Ci₈H₂₄INO₄S + H)⁺: 478 MS found (electrospray): (M+H)⁺ = 478

Intermediate 41

Methyl 5-(4-furo[3,2-fe]pyridin-2-ylphenyl)-3-[[(frans-4- methylcyclohexyl)carbonyl](tetrahydro-3-furanyl)amino]-2-thiophenecarboxylate

To a solution of Intermediate 40a (45 mg) in DME (0.5 mL) and water (0.1 mL) was added 2- [4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl]furo[3,2-ιb]pyridine (45 mg, a synthesis of which is described above as Intermediate 4), potassium phosphate (40 mg) and tetrakis(triphenylphosphine)palladium (0) (1 1 mg). The reaction mixture was stirred at 8O⁰C under nitrogen for 2 h. The mixture was allowed to cool to room temperature and was partitioned between saturated sodium bicarbonate solution and EtOAc. The organics were separated and evaporated in vacuo. The crude material was purified by ISCO Companion silica chromatography, eluting with a gradient 5-100% EtOAc in cyclohexane to give the title compound.

MS calcd for (C_3IH₃₂N₂O₅S + H)⁺: 545 MS found (electrospray): (M+H)⁺ = 545

Intermediate 42 Methyl 3-[(tetrahydro-2H-pyran-4-ylmethyl)amino]-2-thiophenecarboxylate

To a stirred solution of methyl 3-amino-2-thiophenecarboxylate (6.28 g) in DCM (100 ml.) was added glacial acetic acid (7.2 mL) and tetrahydro-2/-/-pyran-4-carbaldehyde (5 g). Sodium triacetoxyborohydride (10.2 g) was added portion-wise and the reaction was stirred at room temperature overnight. A further portion of sodium triacetoxyborohydride (1 g) was added and the reaction mixture was stirred at room temperature for 1 h. Water (100 mL) was added and the reaction was then neutralised with sodium bicarbonate. The DCM layer was separated and the aqueous was extracted with DCM (x 2). The combined organics were dried by passing through a hydrophobic frit and were concentrated in vacuo. The crude material was purified by ISCO Companion silica chromatography, eluting with a gradient 5- 50% EtOAc in cyclohexane to give the title compound. MS calcd for (Ci₂H₁₇NO₃S + H)⁺: 256 MS found (electrospray): (M+H)⁺ = 256

Intermediate 43

Methyl S-IKfrans^-methylcyclohexylJcarbonylKtetrahydro^H-pyran^- ylmethyl)amino]-2-thiophenecarboxylate

Intermediate 42 (2.2 g) and frans-4-methylcyclohexanecarbonyl chloride¹ (1.6 g) were dissolved in DCE (40 mL). The reaction mixture was stirred at 9O⁰C overnight and was then allowed to cool to room temperature. The reaction was quenched with saturated sodium carbonate solution. The organics were separated and the aqueous was washed with DCM

(x 2). The combined organics were dried by passing through a hydrophobic frit and were evaporated in vacuo to give the title compound. MS calcd for (C₂₀H₂₉NO₄S + H)⁺: 380

MS found (electrospray): (M+H)⁺ = 380

Ref 1 : WO2004/052885

Intermediate 44 Methyl S-iodo-S-IKfrans^-methylcyclohexylJcarbonylKtetrahydro^H-pyran^- ylmethyl)amino]-2-thiophenecarboxylate

A solution of LDA (2M solution in THF/heptane/ethyl benzene, 16.5 ml.) was cooled to -78⁰C. A solution of methyl 3-[[(frans-4-methylcyclohexyl)carbonyl](tetrahydro-2/-/-pyran-4- ylmethyl)amino]-2-thiophenecarboxylate (4.2 g, a synthesis of which is described above as Intermediate 43) in dry THF (42 ml.) was added dropwise, keeping the internal temperature between -78⁰C and -7O⁰C and the mixture was stirred at -78⁰C for 15 mins. A solution of iodine (5.6 g) in dry THF (42 ml.) was added dropwise keeping the internal temperature between -78⁰C and -7O⁰C and the reaction mixture was stirred at -78⁰C for 10 mins. The reaction was quenched by dropwise addition of saturated ammonium chloride solution (60 ml.) and was then allowed to warm to room temperature. The reaction mixture was washed with 5% sodium thiosulphate solution and the organics were separated. The aqueous phase was extracted with DCM (x 2) and the combined organics were dried over sodium sulphate and evaporated in vacuo. The crude material was purified by ISCO Companion silica chromatography, eluting with a gradient 5-100% EtOAc in cyclohexane to give the title compound.

MS calcd for (C₂₀H₂₈INO₄S + H)⁺: 506 MS found (electrospray): (M+H)⁺ = 506

Intermediate 45 Methyl 5-(4-imidazo[1,2-a]pyridin-2-ylphenyl)-3-[[(frans-4- methylcyclohexylJcarbonylJ^etrahydro^H-pyran^-ylmethylJamino]^- thiophenecarboxylate

A mixture of Intermediate 44 (200 mg), 2-[4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)phenyl]imidazo[1 ,2-a]pyridine (152 mg, a synthesis of which is described above as Intermediate 7), sodium carbonate (168 mg) in water (2 ml.) and tetrakis(triphenylphosphine)palladium (0) (50 mg) were stirred in DMF (6 ml.) and heated at 100⁰C for 2 h. The reaction was cooled to room temperature and evaporated in vacuo. The residue was partitioned between DCM and water, separated by hydrophobic frit and evaporated in vacuo. The crude material was purified by ISCO Companion silica chromatography, eluting with a gradient 10-100% EtOAc in cyclohexane to give the title compound.

MS calcd for (C₃₃H₃₇N₃O₄S + H)⁺: 572 MS found (electrospray): (M+H)⁺ = 572

Intermediate 46

Methyl S-IKfrans^-methylcyclohexylJcarbonylKtetrahydro^H-pyran^- ylmethyl)amino]-5-(4-pyrazolo[1,5-a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylate

A mixture of Intermediate 44 (200 mg), 2-[4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)phenyl]pyrazolo[1 ,5-a]pyrimidine (152 mg, a synthesis of which is described above as Intermediate 2), sodium carbonate (168 mg) in water (2 ml.) and tetrakis(triphenylphosphine)palladium (0) (50 mg) were stirred in DMF (6 ml.) and heated at 100⁰C for 2.5 h. The reaction was cooled to room temperature and evaporated in vacuo. The residue was partitioned between DCM and water, separated by hydrophobic frit and evaporated in vacuo. The crude material was purified by ISCO Companion silica chromatography, eluting with a gradient 10-100% EtOAc in cyclohexane to give the title compound. MS calcd for (C₃₂H₃₆N₄O₄S + H)⁺: 573 MS found (electrospray): (M+H)⁺ = 573

Intermediate 47 frans-4-(Trifluoromethyl)cyclohexanecarbonyl chloride

Oxalyl chloride (4.59 ml.) was added dropwise to a solution of trans-4- (trifluoromethyl)cyclohexanecarboxylic acid² (6.85 g) in dry DCM (100 ml.) at room temperature under nitrogen. After 10 mins an effervescence was observed and the reaction was stirred at room temperature overnight. The solvent was evaporated in vacuo to give the title compound. 1H NMR (de-DMSO) δ 2.38-2.19 (2H, m), 1.92 (4H, dd), 1.44-1.22 (4H, m). Ref 2: DE 39 30 1 19 (A1 ) Intermediate 48

Methyl S-^Ifrans^^trifluoromethylJcyclohexyllcarbony^amino)^- thiophenecarboxylate

A mixture of methyl 3-amino-2-thiophenecarboxylate (2.93 g), trans-4- (trifluoromethyl)cyclohexanecarbonyl chloride (7.99 g, a synthesis of which is described above as Intermediate 47) and triphenylphosphine (9.78 g) in DCM (12 ml.) was stirred at 5O⁰C under nitrogen overnight. The reaction was evaporated in vacuo and was partitioned between DCM and water. The organic layer was washed with water and the aqueous layer was extracted with DCM. The combined organics were evaporated in vacuo, and the residue was purified by ISCO Companion silica chromatography, eluting with a gradient 0- 100% EtOAc in cyclohexane to give the title compound. MS calcd for (Ci₄H₁₆F₃NO₃S + H)⁺: 336 MS found (electrospray): (M+H)⁺ = 336

Intermediate 49

Methyl 3-([2-(methyloxy)ethyl]{[frans-4-(trifluoromethyl)cyclohexyl]carbonyl}amino)-2- thiophenecarboxylate

A suspension of Intermediate 48 (6.06 g) in dry DMF (200 ml.) under nitrogen was stirred and cooled in ice. 60% Sodium hydride in oil (1.16 g) was added portion-wise over 30 mins. The resulting suspension was allowed to warm to room temperature over 1 h. It was then treated dropwise with 2-bromoethyl methyl ether (5.03 g) in DMF (10 ml_). The resulting solution was heated at 9O⁰C under nitrogen overnight. The reaction was allowed to cool to room temperature, was treated with MeOH and was evaporated in vacuo. The residue was partitioned between DCM and water and the aqueous layer was extracted with DCM. The combined organics were evaporated in vacuo and the crude material was purified by ISCO Companion silica chromatography, eluting with a gradient 0-100% EtOAc in cyclohexane to give the title compound. MS calcd for (Ci₇H₂₂F₃NO₄S + H)⁺: 394 MS found (electrospray): (M+H)⁺ = 394

Intermediate 50

Methyl 5-iodo-3-([2-(methyloxy)ethyl]{[frans-4-

(trifluoromethyl)cyclohexyl]ca carboxylate

A solution of LDA (2M solution in THF/heptane/ethyl benzene, 7 mL) was cooled to -79⁰C under nitrogen. A solution of Intermediate 49 (1.8 g) in dry THF (14 mL) was added slowly over 10 mins. After stirring at -75⁰C for 1.75 h, a solution of iodine (2.31 g) in dry THF (10 mL) was added dropwise over 10 mins, keeping the internal temperature below -7O⁰C. After 45 mins the mixture was quenched with saturated ammonium chloride solution (15 mL). The mixture was allowed to warm to O⁰C and was then washed with 5% sodium thiosulphate solution (50 mL). The organic phase was separated and evaporated in vacuo. The crude material was partitioned between DCM and water and the aqueous was extracted with DCM (x 2). The combined organics were evaporated in vacuo and the residue was purified by ISCO Companion silica chromatography, eluting with a gradient 0-60% EtOAc in cyclohexane. The material obtained was purified further by ISCO Companion silica chromatography, eluting with a gradient 0-25% EtOAc in cyclohexane to give the title compound.

MS calcd for (Ci₇H₂₁F₃INO₄S + H)⁺: 520 MS found (electrospray): (M+H)⁺ = 520

Intermediate 51 Methyl 3-([2-(methyloxy)ethyl]{[frans-4-(trifluoromethyl)cyclohexyl]carbonyl}amino)-5- (4-pyrazolo[1,5-a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylate

A mixture of Intermediate 50 (150 mg), 2-[4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)phenyl]pyrazolo[1 ,5-a]pyrimidine (93 mg, a synthesis of which is described above as Intermediate 2), sodium carbonate (123 mg) in water (0.5 ml.) and tetrakis(triphenylphosphine)palladium (0) (33 mg) was stirred in 1 ,4-dioxane (1.5 ml.) and heated at 100⁰C in a microwave for 40 mins. The reaction was evaporated in vacuo and the residue was partitioned between DCM and water. The aqueous layer was extracted with some more DCM and the organics were combined and evaporated in vacuo. The crude material was purified by ISCO Companion silica chromatography, eluting with a gradient 0- 100% EtOAc in cyclohexane to give the title compound. MS calcd for (C₂₉H₂₉F₃N₄O₄S + H)⁺: 587 MS found (electrospray): (M+H)⁺ = 587

Intermediate 52

Methyl 5-(4-furo[3,2-b]pyridin-2-ylphenyl)-3-([2-(methyloxy)ethyl]{[frans-4-

(trifluoromethyl)cyclohexyl]carbonyl}amino)-2-thiophenecarboxylate

A mixture of 2-[4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl]furo[3,2-;b]pyridine (96 mg, a synthesis of which is described above as Intermediate 4), Intermediate 50 (155 mg), sodium carbonate (127 mg) in water (0.5 ml.) and tetrakis(triphenylphosphine)palladium (0) (35 mg) were stirred in 1 ,4-dioxane (1.5 ml.) and then heated at 100⁰C in a microwave for 35 mins. The reaction was evaporated in vacuo and the residue was partitioned between DCM and water. The aqueous layer was extracted with some more DCM and the organics were combined and evaporated in vacuo. The crude material was purified by ISCO Companion silica chromatography, eluting with a gradient of EtOAc in cyclohexane to give the title compound. MS calcd for (C₃₀H₂₉F₃N₂O₅S + H)⁺: 587 MS found (electrospray): (M+H)⁺ = 587

Intermediate 53

Methyl 5-(4-imidazo[2,1 -fe][1,3]thiazol-6-ylphenyl)-3-[[(frans-4- methylcyclohexylJcarbonylJ^etrahydro^H-pyran^-ylmethylJamino]^- thiophenecarboxylate

A mixture of Intermediate 44 (200 mg), 6-[4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)phenyl]imidazo[2,1-t)][1 ,3]thiazole (154 mg, a synthesis of which is described above as Intermediate 8), sodium carbonate (168 mg) in water (2 ml.) and tetrakis(triphenylphosphine)palladium (0) (50 mg) in DMF (6 ml.) was stirred at 100⁰C for 2 h. The reaction was allowed to cool to room temperature and was evaporated in vacuo. The residue was partitioned between DCM and water, the organics were separated by hydrophobic frit and evaporated in vacuo. The crude material was purified by ISCO Companion silica chromatography, eluting with a gradient 5-100% EtOAc in cyclohexane to give the title compound. MS calcd for (C₃IH₃₅N₃O₄S₂ + H)⁺: 578 MS found (electrospray): (M+H)⁺ = 578

Intermediate 54

Methyl 3-(tetrahydro-2H-pyran-4-yl{[frans-4-

(trifluoromethyl)cyclohexyl]carbonyl}amino)-2-thiophenecarboxylate

To a solution of methyl 3-(tetrahydro-2/-/-pyran-4-ylamino)-2-thiophenecarboxylate (500 mg, a synthesis of which is described above as Intermediate 12) and triphenylphosphine (1.09 g) in dry DCM (2 ml.) was added frans-4-(trifluoromethyl)cyclohexanecarbonyl chloride (666 mg, a synthesis of which is described above as Intermediate 47). The mixture was stirred under nitrogen at 5O⁰C overnight. A further portion of trans-4-

(trifluoromethyl)cyclohexanecarbonyl chloride (50 mg, a synthesis of which is described above as Intermediate 47) was added and the reaction was stirred for 3 h. The reaction mixture was partitioned between water and DCM. The aqueous layer was extracted with DCM, and the combined organics were evaporated in vacuo. The crude material was purified by ISCO Companion silica chromatography, eluting with a gradient 0-50% EtOAc in cyclohexane to give the title compound. MS calcd for (Ci₉H₂₄F₃NO₄S + H)⁺: 420 MS found (electrospray): (M+H)⁺ = 420 Intermediate 55

Methyl 5-iodo-3-(tetrahydro-2H-pyran-4-yl{[ frans-4-

(trifluoromethyl)cyclohexyl]carbonyl}amino)-2-thiophenecarboxylate

A solution of LDA (2M solution in THF/heptane/ethyl benzene, 3 ml.) was cooled to -79⁰C under nitrogen. A solution of Intermediate 54 (816 mg) in dry THF (9 ml.) was added slowly over 10 mins, keeping the internal temperature between -8O⁰C and -7O⁰C. After stirring at - 73⁰C for 1.75 h, a solution of iodine (1.0 g) in dry THF (10 ml.) was added dropwise over 10 mins, keeping the internal temperature below -7O⁰C. After 45 mins the mixture was quenched with saturated ammonium chloride solution (5 ml_). The mixture was allowed to warm to O⁰C and was then washed with 5% sodium thiosulphate solution (25 ml_). The organics were separated and evaporated in vacuo. The residue was partitioned between DCM and water, the aqueous layer was separated using a hydrophobic frit and was extracted with DCM (x 2). The combined organics were evaporated in vacuo. The crude material was purified by ISCO Companion silica chromatography, eluting with a gradient 0- 60% EtOAc in cyclohexane to give the title compound. MS calcd for (Ci₉H₂₃F₃INO₄S + H)⁺: 546 MS found (electrospray): (M+H)⁺ = 546

Intermediate 56

Methyl 5-(4-pyrazolo[1,5-a]pyrimidin-2-ylphenyl)-3-(tetrahydro-2H-pyran-4-yl{[frans-4-

(trifluoromethyl)cyclohexyl]carbonyl}amino)-2-thiophenecarboxylate

A mixture of Intermediate 55 (250 mg), 2-[4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)phenyl]pyrazolo[1 ,5-a]pyrimidine (221 mg, a synthesis of which is described above as Intermediate 2), sodium carbonate (146 mg) in water (0.7 ml.) and tetrakis(triphenylphosphine)palladium (0) (53 mg) in 1 ,4-dioxane (2 ml.) was stirred at 100⁰C in a microwave for 10 mins. The reaction was evaporated in vacuo and the residue was partitioned between water and DCM. The organics were washed with water and the aqueous was extracted with DCM. The combined organics were evaporated in vacuo. The crude material was purified by ISCO Companion silica chromatography, eluting with a gradient 0-100% EtOAc in cyclohexane followed by MeOH. A solid precipitated from the MeOH fractions and this was filtered off and dried in an oven to give the title compound. MS calcd for (C31H31 F₃N₄O₄S + H)⁺: 613 MS found (electrospray): (M+H)⁺ = 613

Intermediate 57 Methyl 5-(4-furo[3,2-b]pyridin-2-ylphenyl)-3-(tetrahydro-2H-pyran-4-yl{[frans-4- (trifluoromethyl)cyclohexyl]carbonyl}amino)-2-thiophenecarboxylate

A mixture of Intermediate 55 (250 mg), 2-[4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)phenyl]furo[3,2-t)]pyridine (221 mg, a synthesis of which is described above as Intermediate 4), sodium carbonate (146 mg) in water (0.7 ml.) and tetrakis(triphenylphosphine)palladium (0) (53 mg) in 1 ,4-dioxane (2 ml.) was stirred at 100⁰C in a microwave for 15 mins. The reaction was evaporated in vacuo and the residue was partitioned between water and DCM. The organics were washed with water and the aqueous was extracted with DCM. The combined organics were evaporated in vacuo. The crude material was purified by ISCO Companion silica chromatography, eluting with a gradient 0-100% EtOAc in cyclohexane followed by MeOH to give the title compound. MS calcd for (C₃₂H_3IF₃N₂O₅S + H)⁺: 613 MS found (electrospray): (M+H)⁺ = 613

Intermediate 58

Methyl 5-[4-(7-aminopyrazolo[1,5-a]pyrimidin-2-yl)phenyl]-3-(tetrahydro-2H-pyran-4- yl{[frans-4-(trifluoromet henecarboxylate

A mixture of Intermediate 55 (100 mg), 2-[4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)phenyl]pyrazolo[1 ,5-a]pyrimidin-7-amine (61.5 mg, a synthesis of which is described above as Intermediate 6), sodium carbonate (58 mg) in water (0.5 ml.) and tetrakis(triphenylphosphine)palladium (0) (21 mg) in 1 ,4-dioxane (1.5 ml.) was stirred at 100⁰C in a microwave for 30 mins. The reaction was evaporated in vacuo and the residue was partitioned between water and DCM. The aqueous layer was extracted with DCM and the organics were washed with water. The combined organics were evaporated in vacuo and the residue was purified by ISCO Companion silica chromatography, eluting with a gradient 0-100% EtOAc in cyclohexane to give the title compound. MS calcd for (C_3IH₃₂F₃N₅O₄S + H)⁺: 628 MS found (electrospray): (M+H)⁺ = 628

Intermediate 59 2-(4-Bromophenyl)[1,3]oxazolo[4,5-ϋ]pyridine

A mixture of 2-amino-3-pyridinol (3 g), 4-bromobenzoic acid (10.96 g) and polyphosphoric acid (7.5 g) were heated at 185⁰C and this was stirred for 30 mins. The reaction was cooled slightly, then ice/water was added. The mixture was filtered and the filtrate was neutralised with saturated sodium bicarbonate solution. The solution was extracted with DCM (x 3), and the combined organics were dried over magnesium sulphate and were evaporated in vacuo.

The crude material was purified by ISCO Companion silica chromatography, eluting with a gradient 0-100% EtOAc in cyclohexane.

MS calcd for (Ci₂H₇BrN₂O + H)⁺: 275/277

MS found (electrospray): (M+H)⁺ = 275/277

Intermediate 60

2-[4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl][1,3]oxazolo[4,5-fe]pyridine

A mixture of Intermediate 59 (2.64 g), bis(pinacolato)diboron (3.66 g), potassium acetate (2.83 g) and PdCI₂(dppf) (518 mg) were dissolved in dry 1 ,4-dioxane (50 ml.) and were heated at 100⁰C under nitrogen for 1 h. The reaction was evaporated in vacuo and the residue was partitioned between water and DCM. The aqueous was extracted with DCM (x

3), then the combined organics were washed with brine, dried over magnesium sulphate and evaporated in vacuo. The crude material was purified by ISCO Companion silica chromatography, eluting with a gradient 0-100% EtOAc in cyclohexane to give the title compound.

MS calcd for (Ci₈H₁₉BN₂O₃ + H)⁺: 323 MS found (electrospray): (M+H)⁺ = 323 Intermediate 61

Methyl 3-{[1 -methyl-2-(methyloxy)ethyl]amino}-2-thiophenecarboxylate

NH

O To a solution of methyl 3-amino-2-thiophenecarboxylate (2 g) and acetic acid (1 ml.) in dry

DCM (25 mL) was added methoxyacetone (2.35 mL) and the reaction was heated at reflux under nitrogen for 2 h. Sodium triacetoxyborohydride (8.1 g) was added portion-wise and the reaction was stirred at reflux under nitrogen for 4 days. The solvent was evaporated in vacuo and the residue was taken into water (100 mL) and was extracted with DCM (3 x 50 mL). The organics were separated using a hydrophobic frit and were evaporated in vacuo.

The crude material was purified by ISCO Companion silica chromatography eluting with a gradient 0-100% EtOAc in cyclohexane to give the title compound.

¹H NMR (d₆-DMSO) δ 7.69 (1 H, d), 6.88 (1 H, d), 3.86-3.76 (1 H, m), 3.71 (3H, d), 3.36 (2H, d), 3.33 (1 H, s), 3.28 (3H, s), 1.15 (3H, d).

Intermediate 62

Methyl S-fl^rans^-methylcyclohexylJcarbonylHI -methyl^-fmethyloxyJethyllamino}^- thiophenecarboxylate

To a solution of Intermediate 61 (0.99 g) and triphenylphosphine (2.27 g) in dry DCM (25 mL) was added frans-4-methylcyclohexanecarbonyl chloride¹ (1.39 g) and the reaction was heated at reflux under nitrogen for 2 days. A further portion of trans-4- methylcyclohexanecarbonyl chloride¹ (0.7 mL) was added and the reaction mixture was heated at reflux under nitrogen for a further 4 days. Sodium bicarbonate solution (50 mL) was added and the organics were extracted with DCM (3 x 50 mL). The combined organics were dried using a hydrophobic frit and were evaporated in vacuo. The crude material was purified by NH₂ SPE cartridge, eluting with MeOH. The material was purified further by ISCO Companion silica chromatography, eluting with a gradient 0-100% EtOAc in cyclohexane to give the title compound. MS calcd for (Ci₈H₂₇NO₄S + H)⁺: 354 MS found (electrospray): (M+H)⁺ = 354 Ref 1 : WO2004/052885

Intermediate 63

Methyl S-iodo-S^^frans^-methylcyclohexylJcarbonylJII -methyl^-

(methyloxy)ethyl]amino}-2-thiophenecarboxylate

A solution of LDA (1.8M solution in THF/heptane/ethyl benzene, 7.1 ml.) in dry THF (10 mL) was cooled to -78⁰C under nitrogen. A solution of Intermediate 62 (1.51 g) in dry THF (10 mL) was added dropwise. The reaction was stirred at -78⁰C under nitrogen for 15 mins, then a solution of iodine (2.18 g) in dry THF (20 mL) was added dropwise. The reaction mixture was stirred under nitrogen at a temperature below -7O⁰C for 2 h, and was then allowed to warm to room temperature. Saturated sodium thiosulphate solution (50 mL) was added and the organics were extracted with EtOAc (x 2). The organics were separated, dried using a hydrophobic frit and evaporated in vacuo. The crude material was purified by ISCO

Companion silica chromatography, eluting with a gradient 0-100% EtOAc in cyclohexane to give the title compound.

MS calcd for (Ci₈H₂₆INO₄S + H)⁺: 480

MS found (electrospray): (M+H)⁺ = 480

Intermediate 64

Methyl S-fl^rans^-methylcyclohexylJcarbonylHI -methyl^-fmethyloxyJethyllamino}^-

(4-pyrazolo[1,5-a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylate

A mixture of Intermediate 63 (96 mg), 2-[4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)phenyl]pyrazolo[1 ,5-a]pyrimidine (64 mg, a synthesis of which is described above as Intermediate 2), sodium carbonate (63 mg) and tetrakis(triphenylphosphine)palladium (0) (23 mg) in 1 ,4-dioxane (1.5 mL) and water (0.5 mL) was heated in a microwave at 100⁰C for 10 mins. The reaction mixture was taken into water (40 mL), extracted with EtOAc (50 mL), separated and dried using a hydrophobic frit. The crude material was purified by ISCO

Companion silica chromatography, eluting with a gradient 0-100% EtOAc in cyclohexane to give the title compound.

MS calcd for (C₃₀H₃₄N₄O₄S + H)⁺: 547

MS found (electrospray): (M+H)⁺ = 547

Intermediate 65

Methyl 5-(4-imidazo[1,2-a]pyridin-2-ylphenyl)-3-(tetrahydro-2H-pyran-4-yl{[frans-4-

(trifluoromethyl)cyclohexyl]carbonyl}amino)-2-thiophenecarboxylate

A mixture of 2-(4-bromophenyl)imidazo[1 ,2-a]pyridine^':i (75.1 mg), bis(pinacolato)diboron (70 mg), potassium acetate (81 mg) and 1 ,1 '-bis (diphenylphosphino)ferrocene dichloro palladium (II) (22 mg) in dry 1 ,4-dioxane (1 ml.) was heated in a microwave at 12O⁰C for 35 mins. The solvent was evaporated and the residue was partitioned between water and DCM. The aqueous phase was extracted further with DCM and the combined organics evaporated to give 2-[4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl]imidazo[1 ,2- a]pyridine. To this was added Intermediate 55 (150 mg), sodium carbonate (87.5 mg) in water (0.8 ml_), tetrakis(triphenylphosphine)palladium (0) (32 mg) and 1 ,4-dioxane (2 ml_). The mixture was stirred at 100⁰C in a microwave for 1 h 25 mins. The reaction was evaporated in vacuo and the residue was partitioned between water and DCM. The aqueous layer was extracted with DCM and the organics were washed with water. The combined organics were evaporated in vacuo and the residue was purified by ISCO Companion silica chromatography, eluting with a gradient 0-100% EtOAc in cyclohexane to give the title compound. MS calcd for (C₃₂H₃₂F₃N₃O₄S + H)⁺: 612 MS found (electrospray): (M+H)⁺ = 612

Ref 3: Burkholder, Conrad; Dolbier, William R.; Medebielle, Maurice; Ait-Mohand, Samia, Tetrahedron Lett, 42, 17, 2001 , 3077 - 3080.

Example 1

S-IKfrans^-MethylcyclohexylJcarbonyllttetrahydro-S-furanylJaminol-S^-pyrazoloII.S- a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylic acid

Intermediate 15 (73 mg) and lithium hydroxide solution (2M, 3 ml.) in THF (4 ml.) and EtOH (4 ml.) were stirred at room temperature for 20 h. The mixture was evaporated and partitioned between 2M HCI (3 ml.) and DCM. The organic layer was dried through a hydrophobic frit and evaporated. The residue was by purified by ISCO Companion C18 chromatography eluting with water (containing 0.1% formic acid) then a gradient of 45-95% acetonitrile (containing 0.05% formic acid) to give the title compound. MS calcd for (C₂₉H₃₀N₄O₄S+ H)⁺: 531 MS found (electrospray): (M+H)⁺ = 531 1H NMR (CDCI₃): 5 8.78 (1 H, d),8.52 (1 H, m), 8.06 (2H, m), 7.75 (2H, m), 7.06 (1 H, s), 6.87 (1 h, m), 5.07 (1 H, m), 4.07 (1 H, m), 3.80 (1 H, m), 3.81-3.56 (3H, m), 2.26 (1 H, m), 2.0-1.15 (9H, m), 0.76 (3H, d), 0.73-0.53 (2H, m), carboxylic acid proton not seen.

Example 2 S-IKfrans^-MethylcyclohexylJcarbonyllttetrahydro^H-pyran^-ylJaminol-S^- pyrazolo[1,5-a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylic acid

2-[4-(4,4,5,5-Tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl]pyrazolo[1 ,5-a]pyrimidine (a synthesis of which is described above as Intermediate 2) (183 mg), Intermediate 14 (200 mg), sodium carbonate (173 mg) and tetrakis(triphenylphosphine) palladium(O) (47 mg) in DMF (5 ml.) were heated at 100°C under a nitrogen atmosphere for 16 h. The solvent was removed and the mixture was partitioned between saturated ammonium chloride and DCM. The organic phase was separated and evaporated to give the crude ester. This was dissolved in THF (2 ml_), ethanol (2 ml.) and 2M lithium hydroxide (2 ml.) and stirred at room temperature for 16 h. The reaction mixture then treated with 2N hydrochloric acid (2 ml_), followed by water and dichloromethane. The layers were stirred for 0.5 h, separated by using a hydrophobic frit and concentrated to give the crude product. This was by purified by ISCO Companion C18 chromatography eluted with water (containing 0.1 % formic acid) then a gradient of 30% to 95% acetonitrile (containing 0.05% formic acid) to give the title compound.

MS calcd for (C₃₀H₃₂N₄O₃S+ H)⁺: 545 MS found (electrospray): (M+H)⁺ = 545 ¹H NMR (CDCI₃): 68.77 (1H, dd),8.52 (1H, dd), 8.11 (2H, d), 7.79(2H₁OXy-Iy(IH₁S)₁ZOS (1H₁ S)₁6.8y (1H₁ m), 4.89 (1H₁ m), 4.01 (1H₁ m), 3.93 (1H₁ m), 3.58-3.45 (3H₁ m), 2.10 (1H₁ m),1.96 (1H₁ m), 1.82-1.26 (10H₁ m), 079 (3H₁ d), 076-0.53 (2H₁ m), carboxylic acid proton not seen.

The following examples were similarly prepared.

Example 3

S^-FuroIS^-felpyridin^-ylphenylJ-S-IKfrans^-methylcyclohexylJcarbonyllttetrahydro- 2H-pyran-4-yl)amino]-2-thiophenecarboxylic acid

Prepared from Intermediate 14 (200 mg) and 2-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2- yl)phenyl]furo[3,2-b]pyridine (0.5y mmol; a synthesis of which is described above as Intermediate 4). MS calcd for (C_3IH₃₂N₂O₅S+ H)⁺: 545 MS found (electrospray): (M+H)⁺= 545

¹HNMR(CDCI₃): δ 8.60 (1H₁ dd), 8.06 (2H₁ d), 7.91 (1H₁ dd), 7.82 (2H₁ d),7.48 (1H₁S)₁ 7.34 (1H₁ dd), 7.19 (1H, s), 4.93 (1H, bt), 4.02 (1H, bdd),3.93 (1H, bdd), 3.52 (2H, bt), 2.15 (1H, bt),1.96 (1H, bd), 1.88-1.27 (10H, m), 0.79 (3H, d), 0.8-0.6 (2H, m), carboxylic acid proton not seen.

Example 4

S-^^-AminopyrazoloII.S-alpyrimidin^-ylJphenyll-S-IKfrans^-methylcyclohexyl) carbonylJ^etrahydro^H-pyran^-ylJaminol^-thiophenecarboxylic acid

Prepared from Intermediate 14 (200 mg) and 2-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2- yl)phenyl]pyrazolo[1,5-a]pyrimidin-7-amine (0.57 mmol; a synthesis of which is described above as Intermediate 6). MS calcd for (C₃₀H₃₃N₅O₄S+ H)⁺: 560 MS found (electrospray): (M+H)⁺ = 560 ¹H NMR (CD₃OD): 58.12 (2H, d), 8.05 (1H, d), 7.81 (2H, d), 7.36 (1H, s), 6.8 (1H, s), 6.17 (1H, d), 4.7 (1H, bt), 3.95 (1H, bdd), 3.88 (1H, bdd), 3.48 (2H, m), 2.17 (1H, tt), 1.95-1.12 (11 H, m), 0.77 (3H, bd), 0.73-0.55 (2H, m), carboxylic acid proton not seen.

Example 5

S^-lmidazoII^-alpyridin^-ylphenylJ-S-IKfrans^-methylcyclohexylJcarbonyl] (tetrahydro-2H-pyran-4-yl)amino]-2-thiophenecarboxylic acid

Prepared from Intermediate 14 (200 mg) and 2-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2- yl)phenyl]imidazo[1,2-a]pyridine (0.57 mmol; a synthesis of which is described above as

Intermediate 7).

MS calcd for (C₃i H₃₃N₃O₄S+ H)⁺: 545

MS found (electrospray): (M+H)⁺= 545

¹H NMR (CDCI₃): 58.16 (1H, d),8.02 (2H, bd), 7.90 (1H, s), 7.87 (1H, bd), 7.78 (2H, d), 7.28 (1H,bt), 7.15 (1H, s), 6.88 (1H, bt), 4.88 (1H, bt), 3.99 (1H, bdd), 3.87 (1H, bdd), 3.51(1H, bt),

3.43 (1H,bt), 2.17 (1H, bt),1.93 (1H, bd), 1.80 (1H,bd), 1.73-1.25 (10H, d), 0.775 (3H, m), 0.8-0.6 (2H, m), carboxylic acid proton not seen.

Example 6 S^-lmidazo^.i-felli.Slthiazol-θ-ylphenylJ-S-IKfrans^-methylcyclohexyOcarbonyl] (tetrahydro-2H-pyran-4-yl)amino]-2-thiophenecarboxylic acid

Prepared from Intermediate 14 (200 mg) and 6-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2- yl)phenyl]imidazo[2,1-b][1,3]thiazole (0.57 mmol; a synthesis of which is described above as Intermediate 8).

MS calcd for (C₂₉H₃₁N₃O₄S₂+ H)⁺: 549

MS found (electrospray): (M+H)⁺= 549

¹H NMR (DMSO): 58.31 (1H, s), 7.92 (1H,d), 7.9 (2H, d), 7.8 (2H, d), 7.53 (1H, bs), 7.26

(1H,d), 4.55 (1H, m), 3.82 (1H, bd), 3.76 (1H, bd), 3.4-3.3 (2H, m), 1.95 (1H, bt), 1.75 (1H, bd), 1.65-1.05 (1 OH, m), 0.7 (3H, d), 0.65-0.46 (2H, m), carboxylic acid proton not seen.

Example 7 S^-FuroIS^-felpyridin^-ylphenylJ-S^Kfrans^-methylcyclohexylJcarbonyl]^- (methyloxy)ethyl]amino}-2-thiophenecarboxylic acid

A solution of Intermediate 18 (0.302 g) in THF (2.5 ml.) and MeOH (2.5 ml.) was stirred and treated with 2N sodium hydroxide (1.5 ml.) for 2 h. The reaction was evaporated and the mixture partitioned between water and EtOAc. The aqueous layer was separated and acidified to pH2 with 2N HCI and extracted with EtOAc. The organic phase was washed with brine, dried over sodium sulphate, and evaporated. This was then purified by ISCO Companion silica chromatography eluting with a gradient of EtOAc in cyclohexane (0% to 20%). Combined fractions of the product were evaporated and placed on a silica SPE column and eluted with EtOAc/cyclohexane, DCM, and finally DCM/MeOH to give the title compound.

MS calcd for (C₂₉H₃₀SN₂O₅+ H)⁺: 519 MS found (electrospray): (M+H)⁺ = 519 1H NMR (CD₃OD): 5 8.47 (1 H, bd), 8.06 (2H, bd), 8.01 (2H, bd), 7.87 (2H, d), 7.53 (1 H, s), 7.43 (1 H, s), 7.37 (1 H, dd), 4.17 (1 H, m), 3.64-3.3.47 (3H, m), 3.3 (3H, s), 2.28 (1 H, bt), 1.79 (2H, bd), 1.66 (2H, bd), 1.5 (2H, m), 1.31 (1 H, bm), 0.8 (3H, d), 0.71 (2H, m), carboxylic acid proton not seen.

Example 8

5-[4-(7-Aminopyrazolo[1,5-a]pyrimidin-2-yl)phenyl]-3-{[(frans-4- methylcyclohexyl)carbonyl][2-(methyloxy)ethyl]amino}-2-thiophenecarboxylic acid

A solution of Intermediate 19 (0.45g) in THF (3 ml.) and MeOH (3 ml.) was stirred and treated with 2N sodium hydroxide (2.5 ml.) for 2 h. The reaction was evaporated and the mixture partitioned between water and EtOAc. The aqueous was separated and acidified to pH2 with 2N HCI and extracted with EtOAc. The organic phase was washed with brine, dried over sodium sulphate, and evaporated. The residue was purified on a silica SPE column and eluted with EtOAc/cyclohexane, DCM, and finally DCM/MeOH to give the title compound.

MS calcd for (C₂₈H₃₁SN₅O₄+ H)⁺: 534 MS found (electrospray): (M+H)⁺ = 534 ¹H NMR (CD₃OD): 5 8.1 1 (2H, d), 8.05 (1 H, d), 7.79 (2H, d), 7.45 (1 H,s), 6.78 (1 H, s), 6.17 (1 H,d), 4.2 (1 H, m), 3.65-3.46 (3H, m), 3.3 (3H,s), 2.3 (1 H, tt), 1.79 (2H, bm), 1.65 (2H, bd), 1.49 (2H, m), 1.30 (1 H, m), 0.79 (3H, d), 0.71 (2H, m), carboxylic acid proton not seen.

Example 9

3-{[(frans-4-Methylcyclohexyl)carbonyl][2-(methyloxy)ethyl]amino}-5-(4-pyrazolo[1,5- a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylic acid

A solution of Intermediate 20 (0.12 g) in THF (2 ml.) and MeOH (2 ml.) was stirred and treated with 2N sodium hydroxide (0.6 ml.) for 2 h. The reaction was evaporated and the mixture partitioned between water and EtOAc. The aqueous layer was separated and acidified to pH2 with 2N HCI and extracted with EtOAc. The organic phase was washed with brine, dried over sodium sulphate, and evaporated. The residue was purified on a silica SPE column and eluted with EtOAc/cyclohexane, DCM, and finally DCM/MeOH to give the title compound.

MS calcd for (C₂₈H₃₀SN₄O₄+ H)⁺: 519 MS found (electrospray): (M+H)⁺ = 519

¹H NMR (CD₃OD): 5 8.89 (1 H, bd), 8.46 (1 H, dd), 8.03 (2H, d), 7.76 (2H, d), 7.40 (1 H,s), 7.02 (1 H, s), 6.95 (1 H, dd), 4.27 (1 H, m), 3.63-3.3.44 (3H, m), 3.3 (3H, s), 2.33 (1 H, bt), 1.90-1.74 (2H, bd), 1.625 (2H, bt), 1.47 (2H, m), 1.29 (1 H, m), 0.77 (3H, d), 0.68 (2H, bm), carboxylic acid proton not seen.

Example 10

S^-lmidazoII^-alpyridin^-ylphenylJ-S^Kfrans^-methylcyclohexylJcarbonyl]^- (methyloxy)ethyl]amino}-2-thiophenecarboxylic acid

A solution of Intermediate 21 (0.47 g) in THF (4 ml.) and MeOH (4 ml.) was stirred and treated with 2N sodium hydroxide (2.3 ml.) for 3 h. The reaction was evaporated and the mixture partitioned between water and EtOAc. The aqueous layer was separated and acidified to pH2 with 2N HCI and extracted with EtOAc. The organic phase was washed with brine, dried over sodium sulphate, and evaporated. The residue was purified on a silica SPE column and eluted with EtOAc/cyclohexane, DCM, and finally DCIWMeOH to give the title compound.

MS calcd for (C29H31SN3O4+ H)⁺: 518 MS found (electrospray): (M+H)⁺ = 518 1H NMR (CD₃OD): δ 8.47 (1 H, d), 8.31 (1 H, s), 8.03 (2H, d), 7.83 (2H, d), 7.60 (1 H, bd), 7.49 (1 H, s), 7.38 (1 H, dd), 6.97 (1 H, t), 4.15 (1 H, m), 3.63-3.3.46 (3H, m), 3.3 (3H, s), 2.28 (1 H, bt), 1.78 (2H, bd), 1.67 (2H, bd), 1. 5 (2H, m), 1.31 (1 H, m), 0.81 (3H, d), 0.73 (2H, m), carboxylic acid proton not seen.

Example 11

S^-lmidazo^.i -feHI.Slthiazol-e-ylphenylJ-S^Kfrans^-methylcyclohexyOcarbonyl]^- (methyloxy)ethyl]amino}-2-thiophenecarboxylic acid

A solution of Intermediate 22 (0.43 g) in THF (3.5 ml.) and MeOH (3.5 ml.) was stirred and treated with 2N sodium hydroxide (2 ml.) for 3 h. The reaction was evaporated and the mixture partitioned between water and EtOAc. The aqueous layer was separated and acidified to pH2 with 2N HCI and extracted with EtOAc. The organic phase were washed with brine, dried over sodium sulphate, and evaporated. The residue was purified on a silica

SPE column and eluted with EtOAc/cyclohexane, DCM, and finally DCM/MeOH to give the title compound.

MS calcd for (C₂₇H₂₉S₂N₃O₄+ H)⁺: 524 MS found (electrospray): (M+H)⁺ = 524

¹H NMR (CD₃OD): 5 8.15 (1 H, s), 7.91 (2H, d), 7.81-7.76 (3H, 2xd), 7.47 (1 H, s), 7.18 (1 H, d), 4.14 (1 H, m), 3.65-3.3.47 (3H, 2xm), 3.3 (2H, s), 2.28 (1 H, tt), 1.77 (2H, bt), 1.67 (2H, bd), 1.5 (2H, m), 1.32 (1 H, m), 0.8 (3H, d), 0.73 (2H, m), carboxylic acid proton not seen.

Example 12

3-{[2-(Ethyloxy)ethyl][(frans-4-methylcyclohexyl)carbonyl]amino}-5-(4-furo[3,2- fe]pyridin-2-ylphenyl)-2-thiophenecarboxylic acid

A solution of Intermediate 25 (0.54g) in THF (4 mL) and MeOH (4 mL) was stirred and treated with 2N sodium hydroxide (2.5 mL) for 4 h. The reaction was evaporated and the mixture partitioned between water and EtOAc. The aqueous layer was separated and acidified to pH2 with 2N HCI and extracted with EtOAc. The organic phase was washed with brine, dried over sodium sulphate, and evaporated. The residue was purified on a silica SPE column and eluted with DCM in MeOH (0% to 75%) to give the title compound. MS calcd for (C₃₀H₃₂SN₂O₅+ H)⁺: 533 MS found (electrospray): (M+H)⁺ = 533

¹H NMR (CD₃OD): 5 8.48 (1 H, d), 8.08 (2H, bd), 8.01 (1 H, d), 7.88 (2H, bd), 7.57 (1 H, s), 7.45 (1 H, s), 7.37 (1 H, dd), 4.10 (1 H, m), 3.67-3.52 (3H, m), 3.42 (2H, m), 2.26 (1 H, tt), 1.80- 1.60 (4H, m), 1.48 (2H, m), 1.29 (1 H, bm), 1.11 (3H, t), 0.79 (3H, d), 0.70 (2H, m), carboxylic acid proton not seen.

Example 13 5-[4-(7-Aminopyrazolo[1 ,5-a]pyrimidin-2-yl)phenyl]-3-{[2-(ethyloxy)ethyl][(frans-4- methylcyclohexyl)carbonyl]amino}-2-thiophenecarboxylic acid

A solution of Intermediate 26 (0.43g) in THF (4 mL) and MeOH (4 mL) was stirred and treated with 2N sodium hydroxide (2 mL) for 4 h. The reaction was evaporated and the mixture partitioned between water and EtOAc. The aqueous layer was separated and acidified to pH2 with 2N HCI and extracted with EtOAc. The organic phase was washed with brine, dried over sodium sulphate, and evaporated. The residue was purified on a silica SPE column and eluted with DCM in MeOH (0% to 30%) to give the title compound. MS calcd for (C₂₉H₃₃SN₅O₄+ H)⁺: 548 MS found (electrospray): (M+H)⁺ = 548

¹H NMR (CD₃OD): 5 8.1 1 (2H, d), 8.05 (1 H, d), 7.79 (2H, d), 7.47 (1 H,s), 6.80 (1 H, s), 6.17 (1 H,d), 4.15 (1 H, m), 3.68-3.50 (3H, m), 3.42 (2H, m), 3.3 (3H,s), 2.28 (1 H, tt), 1.78 (2H, bm), 1.65 (2H, bd), 1.49 (2H, m), 1.30 (1 H, bm), 1.12 (3H, t), 0.79 (3H, d), 0.70 (2H, bm), carboxylic acid proton not seen.

Example 14

3-{[2-(Ethyloxy)ethyl][(frans-4-methylcyclohexyl)carbonyl]amino}-5-(4-pyrazolo[1,5- a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylic acid

A solution of Intermediate 27 (0.54 g) in THF (4 mL) and MeOH (4 mL) was stirred and treated with 2N sodium hydroxide (2.5 mL) for 4 h. The reaction was evaporated and the mixture partitioned between water and EtOAc. The aqueous layer was separated and acidified to pH2 with 2N HCI and extracted with EtOAc. The organic phase was washed with brine, dried over sodium sulphate, and evaporated. The residue was purified on a silica SPE column and eluted with DCM in MeOH (0% to 20%) to give the title compound. MS calcd for (C₂₉H₃₂SN₄O₄+ H)⁺: 533 MS found (electrospray): (M+H)⁺ = 533 1H NMR (CD₃OD): 5 8.93 (1 H, dd), 8.51 (1 H, dd), 8.12 (2H, d), 7.84 (2H, bd), 7.53 (1 H, s), 7.12 (1 H, s), 7.01 (1 H, dd), 4.1 1 (1 H, m), 3.68-3.53 (3H, m), 3.42 (2H, m), 2.27 (1 H, tt), 1.76 (2H, bt), 1.65 (2H, bd),1.59-1.376 (2H, m), 1.29, 1 H, bm), 1.12 (3H, t), 0.79 (3H, d), 0.71 (2H, m), carboxylic acid proton not seen.

Example 15

3-{[2-(Ethyloxy)ethyl][(frans-4-methylcyclohexyl)carbonyl]amino}-5-(4-imidazo[1,2- a]pyridin-2-ylphenyl)-2-thiophenecarboxylic acid

A solution of Intermediate 28 (0.696g) in THF (5 mL) and MeOH (5 mL) was stirred and treated with 2N sodium hydroxide (3 mL) for 3 h. The reaction was evaporated and the mixture partitioned between water and EtOA . The aqueous layer was separated and acidified to pH2 with 2N HCI and extracted with EtOA. The organic phase was washed with brine, dried over sodium sulphate, and evaporated. The residue was purified on a silica SPE column and eluted with EtOAc/cyclohexane, EtOAc, DCM, and finally DCM in MeOH(0% to 20%) to give the title compound.

MS calcd for (C₃₀H₃₃SN₃O₄+ H)⁺: 532 MS found (electrospray): (M+H)⁺ = 532

¹H NMR (CD₃OD): 5 8.49 (1 H, bd), 8.34 (1 H, s), 8.01 (2H, d), 7.82 (2H, d), 7.63 (1 H, bd), 7.52 (1 H, bs), 7.45 (1 H, dd), 7.03 (1 H, t), 4.12(1 H, bm), 3.69-3.49 (3H, m), 3.42 (2H, m, 2.27 (1 H, tt), 1.76 (2H, bt), 1.65 (2H, bd), 1.59-1.37 (2H, m),1.29 (1 H, bm), 1.12 (3H, t), 0.79 (3H, d), 0.70 (2H, m), carboxylic acid proton not seen. Example 16

3-{[2-(Ethyloxy)ethyl][(frans-4-methylcyclohexyl)carbonyl]amino}-5-(4-imidazo[2,1 - to] [1 ,3]thiazol-6-ylphenyl)-2-thiophenecarboxylic acid

A solution of Intermediate 29 (0.52 g) in THF (4 mL) and MeOH (4 mL) was stirred and treated with 2N sodium hydroxide (2.36 mL) for 2h. The reaction was evaporated and the mixture partitioned between water and EtOAc. The aqueous layer was separated and acidified to pH2 with 2N HCI and extracted with EtOAc. The organic phase was washed with brine, dried over sodium sulphate, and evaporated. The residue was purified on a silica SPE column eluted with DCM in MeOH (0% to 20%) to give the title compound. MS calcd for (C₂₈H₃IS₂N₃O₄+ H)⁺: 538 MS found (electrospray): (M+H)⁺ = 538

¹H NMR (CD₃OD): 5 8.12 (1 H, s), 7.88 (1 H, d), 7.78- 7.73 (3H, 2xd), 7.47 (1 H, s), 7.16 (1 H, d), 4.11 (1 H, bm), 3.67-3.49 (3H, m), 3.42 (2H, m), 2.26 (1 H, tt), 1.75 (2H, bt), 1.65 (2H, bd), 1.59-1.37 (2H, m), 1.28 (1 H, m), 1.1 1 (3H, t), 0.79 (3H, d), 0.70 (2H, m), carboxylic acid proton not seen.

Example 17

3-([(frans-4-Methylcyclohexyl)carbonyl]{2-(methyloxy)-1 - [(methyloxy)methyl]ethyl}amino)-5-(4-pyrazolo[1,5-a]pyrimidin-2-ylphenyl)-2- thiophenecarboxylic acid

To a solution of Intermediate 36 (191 mg) in THF (4 mL) and ethanol (4 mL) was added 2M lithium hydroxide solution (4 mL). The reaction was stirred at room temperature for 3 days.

The mixture was quenched with 2M HCI (4 mL) and was partitioned between water and

DCM. The layers were separated using a hydrophobic frit and the organics were evaporated in vacuo. The crude material was purified using and NH₂ SPE cartridge, eluting with 10% acetic acid in 1 ,4-dioxane. The material was purified further by ISCO Companion chromatography, using a C18 cartridge, eluting with a gradient 40% MeCN (0.05% formic acid)/water (0.1 % formic acid) to 95% MeCN (0.05% formic acid), and was freeze-dried from 1 ,4-dioxane to give the title compound. MS calcd for (C₃₀H₃₄N₄O₅S + H)⁺: 563 MS found (electrospray): (M+H)⁺ = 563

¹H NMR (CDCI₃) δ 8.75 (1 H, d), 8.52 (1 H, dd), 8.08 (2H, d), 7.77 (2H, d), 7.39 (1 H, s), 7.06 (1 H, d), 6.87 (1 H, dd), 4.79-4.70 (1 H, m), 3.82-3.49 (4H, m), 3.40 (3H, s), 3.30 (3H, s) 2.23- 2.12 (1 H, m), 1.83-1.73 (1 H, m), 1.72-1.55 (4H, m), 1.48-1.24 (2H, m), 0.78 (3H, d), 0.77- 0.59 (2H, m), carboxylic acid proton not seen.

Example 18

S^-FuroIS^-felpyridin^-ylphenylJ-S-IKfrans^-methylcyclohexylJcarbonyllttetrahydro- 3-furanyl)amino]-2-thiophenecarboxylic acid

To a stirred solution of Intermediate 41 (46 mg) in ethanol (2 ml.) and THF (2 ml.) was added 2M lithium hydroxide solution (2 ml.) and the reaction was stirred at room temperature for 16 h. 2M HCI (2 ml.) was added and the mixture was partitioned between water and DCM. The layers were stirred, separated by hydrophobic frit, and evaporated in vacuo. The crude material was purified using an NH₂ SPE cartridge, eluting with 10% acetic acid in 1 ,4-dioxane and the material was then freeze-dried from 1 ,4-dioxane to give the title compound. MS calcd for (C₃₀H₃₀N₂O₅S + H)⁺: 531 MS found (electrospray): (M+H)⁺ = 531

¹H NMR (d₆-DMSO) δ 8.54 (1 H, dd), 8.12-8.07 (3H, m), 7.97 (2H, dd), 7.82-7.75 (2H, m), 7.37 (1 H, dd), 4.94 (1 H, septet), 3.75-3.64 (1 H, m), 3.61-3.44 (4H, m), 2.04-1.87 (2H, m), 1.72-1.15 (7H, m), 0.74 (3H, d), 0.70-0.50 (2H, m), carboxylic acid proton not seen.

Example 19

5-(4-lmidazo[1,2-a]pyridin-2-ylphenyl)-3-[[(frans-4- methylcyclohexylJcarbonyllttetrahydro^H-pyran^-ylmethylJamino]^- thiophenecarboxylic acid

A solution of Intermediate 45 (102 mg) in ethanol (3 ml_), THF (3 mL) and 2N sodium hydroxide solution (1 mL) was stirred at room temperature overnight. DCM (20 mL) was added, followed by 2N HCI (5 mL). The mixture was stirred for 30 mins before the organics were separated by hydrophobic frit. The organics were evaporated in vacuo to give the title compound.

MS calcd for (C₃₂H₃₅N₃O₄S)⁺: 558 MS found (electrospray): (M+H)⁺ = 558

¹H NMR (CD₃OD) δ 8.85 (1 H, d), 8.72 (1 H, s), 8.06-7.93 (6H, m), 7.68-7.49 (2H + impurity, m), 3.95-3.84 (3H, m), 3.43-3.31 (3H, m), 2.23 (1 H, tt), 1.90-1.59 (7H, m), 1.58-1.23 (5H, m), 0.78 (3H, d), 0.74-0.60 (2H, m), carboxylic acid proton not seen.

Example 20

S^-FuroIS^-felpyridin^-ylphenylJ-S-IKfrans^-methylcyclohexylJcarbonyllttetrahydro- 2H-pyran-4-ylmethyl)amino]-2-thiophenecarboxylic acid

A mixture of Intermediate 44 (200 mg), 2-[4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)phenyl]furo[3,2-b]pyridine (152.3 mg, a synthesis of which is described above as Intermediate 4), sodium carbonate (168 mg) in water (2 mL) and tetrakis(triphenylphosphine)palladium (0) (50 mg) were stirred in DMF (6 mL) and heated at 100⁰C overnight. The reaction was evaporated in vacuo and the residue partitioned between DCM and saturated ammonium chloride solution. The organics were separated, dried using a hydrophobic frit and evaporated in vacuo. The crude material was purified by reverse phase ISCO Companion chromatography, using a C18 cartridge, eluting with a gradient 40% MeCN (0.05% formic acid)/water (0.1% formic acid) to 95% MeCN (0.05% formic acid) to give the title compound.

MS calcd for (C₃₂H₃₄N₂O₅S)⁺: 559

MS found (electrospray): (M+H)⁺ = 559

¹H NMR (CD₃OD) δ 8.48 (1 H, dd), 8.10 (2H, d), 8.02 (1 H, d), 7.92 (2H, d), 7.59 (1 H, s), 7.47

(1 H, s), 7.38 (1 H, dd), 3.97-3.82 (3H, m), 3.43-3.32 (2H, m), 2.31-2.20 (1 H, m), 1.89-1.60 (7H, m), 1.58-1.20 (6H, m), 0.79 (3H, d), 0.77-0.66 (2H, m), carboxylic acid proton not seen.

Example 21

S-IKfrans^-MethylcyclohexylJcarbonyllttetrahydro^H-pyran^-ylmethylJaminol-S^- pyrazolo[1,5-a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylic acid

A solution of Intermediate 46 (142 mg) in ethanol (2 ml_), THF (2 mL) and 2N sodium hydroxide solution (1 mL) was stirred at room temperature overnight. DCM (12 mL) was added, followed by 2N HCI (6 mL). The mixture was stirred for 30 mins before the organics were separated by hydrophobic frit. The organics were evaporated in vacuo and purified by an NH₂ SPE cartridge, eluting with 10% 2N HCI in MeOH to give the title compound. MS calcd for (C_3IH₃₄N₄O₄S)⁺: 559 MS found (electrospray): (M+H)⁺ = 559

¹H NMR (CD₃OD) δ 9.15 (1 H, d), 8.65 (1 H, d), 8.15 (2H, d), 7.88 (2H, d), 7.58 (1 H, s), 7.22 (1 H, s), 7.14 (1 H, dd), 3.96-3.84 (3H, m), 3.43-3.30 (2H, m), 2.29-2.20 (1 H, m), 1.90-1.22 (13H, m), 0.79 (3H, d), 0.76-0.63 (2H, m), carboxylic acid proton not seen.

Example 22

3-([2-(Methyloxy)ethyl]{[frans-4-(trifluoromethyl)cyclohexyl]carbonyl}amino)-5-(4- pyrazolo[1,5-a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylic acid

A solution of Intermediate 51 (1 16 mg) in methanol (0.5 mL), THF (0.5 mL) and 2N sodium hydroxide solution (0.5 mL) was stirred at room temperature overnight. The reaction was evaporated in vacuo and partitioned between DCM and water. The aqueous layer was extracted with some more DCM and the organics were combined and evaporated in vacuo. The crude material was purified by an NH₂ SPE cartridge to give the title compound. MS calcd for (C₂₈H₂₇F₃N₄O₄S)⁺: 573 MS found (electrospray): (M+H)⁺ = 573

¹H NMR (CDCI₃) δ 8.74 (1 H, d), 8.52 (1 H, dd), 8.10 (2H, d), 7.77 (2H, d), 7.29 (1 H, s), 7.07 (1 H, s), 6.88 (1 H, dd), 4.04-3.83 (2H, m), 3.71-3.58 (2H, m), 3.35 (3H, s), 2.35-2.22 (1 H, m), 2.08-1.75 (5H, m), 1.74-1.45 (2H, m), 1.24-1.04 (2H, m), carboxylic acid proton not seen.

Example 23 5-(4-Furo[3,2-fe]pyridin-2-ylphenyl)-3-([2-(methyloxy)ethyl]{[frans-4- (trifluoromethyl)cyclohexyl]carbonyl}amino)-2-thiophenecarboxylic acid

A mixture of Intermediate 52 (1 10 mg), THF (0.5 ml_), MeOH (0.5 ml.) and 2N sodium hydroxide solution (0.5 ml.) was stirred at room temperature overnight. The reaction was evaporated in vacuo and the residue was partitioned between DCM and water. The aqueous layer was extracted with DCM, and the combined organics were evaporated in vacuo. The residue was triturated with MeOH to give the title compound. MS calcd for (C₂₉H₂₇F₃N₂O₅S)⁺: 573 MS found (electrospray): (M+H)⁺ = 573

¹H NMR (d₆-DMSO) δ 8.61 (1 H, d), 8.27 (1 H, d), 8.15 (2H, d), 7.99 (2H, d), 7.87 (1 H, s), 7.77 (1 H, s), 1.52-1 Al (1 H, m), 3.96-3.85 (1 H, m, partially obscured by water peak), 3.63-3.53 (1 H, m), 3.51-3.42 (1 H, m), 3.41-3.35 (1 H, m), 3.16 (3H, s), 2.31-2.13 (2H, m), 1.85-1.62 (4H, m), 1.58-1.30 (2H, m), 1.10-0.92 (2H, m), carboxylic acid proton not seen.

Example 24

5-(4-lmidazo[2,1 -fe][1,3]thiazol-6-ylphenyl)-3-[[(frans-4- methylcyclohexylJcarbonyllttetrahydro^H-pyran^-ylmethylJamino]^- thiophenecarboxylic acid

A solution of Intermediate 53 (94 mg) in THF (3 ml_) and ethanol (3 ml.) was treated with 2N sodium hydroxide solution (1 ml.) and the reaction was stirred at room temperature overnight. DCM (20 ml.) was added, followed by 2N HCI (5 ml_). The reaction mixture was stirred for 30mins before being separated by hydrophobic frit and evaporated in vacuo to give the title compound.

MS calcd for (C₃oH33N₃θ₄S₂)⁺: 564 MS found (electrospray): (M+H)⁺ = 564 ¹H NMR (CD₃OD) δ 8.51 (1 H, s), 8.19 (1 H, d), 7.96 (2H, d), 7.89 (2H, d), 7.70 (1 H, d), 7.63 (1 H, s), 3.95-3.83 (3H, m), 3.41-3.32 (2H, m), 2.28-2.18 (1 H, m), 1.89-1.59 (7H, m), 1.59- 1.22 (6H, m), 0.79 (3H, d), 0.75-0.60 (2H, m), carboxylic acid proton not seen.

Example 25

5-(4-Pyrazolo[1,5-a]pyrimidin-2-ylphenyl)-3-(tetrahydro-2H-pyran-4-yl{[frans-4- (trifluoromethyl)cyclohexyl]carbonyl}amino)-2-thiophenecarboxylic acid

Intermediate 56 (168 mg) was dissolved in THF (0.7 ml.) and MeOH (0.7 ml_). 2N Sodium hydroxide solution (0.7 ml.) was added and the mixture was stirred at room temperature overnight. THF (0.5 ml_), 2N sodium hydroxide solution (0.5 ml.) and a few drops of DCM were added and the mixture was stirred at room temperature overnight. The mixture was evaporated in vacuo and the residue was partitioned between DCM and 2M HCI. The aqueous layer was extracted with DCM and the combined organics were evaporated in vacuo. A few drops of diethyl ether were added and the material was dried under nitrogen to give the title compound.

MS calcd for (C₃₀H₂₉F₃N₄O₄S)⁺: 599

MS found (electrospray): (M+H)⁺ = 599

¹H NMR (CDCI₃) δ 8.76 (1 H, d), 8.53 (1 H, dd), 8.12 (2H, d), 7.79 (2H, d), 7.17 (1 H, s), 7.08 (1 H, s), 6.88 (1 H, dd), 4.95-4.83 (1 H, m), 3.98 (2H, dd), 3.60-3.44 (2H, m), 2.22-1.61 (10H, m), 1.59-1.30 (2H, m), 1.22-1.00 (2H, m), carboxylic acid proton not seen.

Example 26

5-(4-Furo[3,2-fe]pyridin-2-ylphenyl)-3-(tetrahydro-2H-pyran-4-yl{[frans-4- (trifluoromethyl)cyclohexyl]carbonyl}amino)-2-thiophenecarboxylic acid formic acid salt

Intermediate 57 (250 mg) was dissolved in THF (0.7 ml.) and MeOH (0.7 ml_). 2N Sodium hydroxide solution (0.7 ml.) was added and the mixture was stirred at room temperature overnight. The reaction was evaporated in vacuo and was partitioned between DCM and 2M HCI. The aqueous was extracted with DCM and the combined organics were evaporated in vacuo. The crude material was purified by MDAP HPLC to give the title compound. MS calcd for (C₃i H₂₉F₃N₂O₅S)⁺: 599 MS found (electrospray): (M+H)⁺ = 599

¹H NMR (CD₃OD) δ 8.46 (1 H, d), 8.15 (1 H, s), 8.06 (2H, d), 8.00 (1 H, d), 7.89 (2H, d), 7.48 (1 H, s), 7.43 (1 H, s), 7.36 (1 H, dd), 4.76-4.66 (1 H, m), 4.00-3.85 (2H, m), 3.56-3.42 (2H, m), 2.29-1.54 (1OH, m), 1.51-1.24 (2H, m), 1.17-0.93 (2H, m), carboxylic acid protons not seen.

Example 27

5-[4-(7-Aminopyrazolo[1,5-a]pyrimidin-2-yl)phenyl]-3-(tetrahydro-2H-pyran-4-yl{[frans- 4-(trifluoromethyl)cyclohexyl]carbonyl}amino)-2-thiophenecarboxylic acid

Intermediate 58 (70 mg) was dissolved in THF (0.3 ml.) and MeOH (0.3 ml_). 2N Sodium hydroxide solution (0.3 ml.) was added and the mixture was stirred at room temperature overnight. The reaction was evaporated in vacuo and was partitioned between DCM and water. The aqueous was extracted with DCM and the combined organics were evaporated in vacuo. The crude material was purified by ion exchange cartridge to give the title compound.

MS calcd for (C₃₀H₃₀F₃N₅O₄S)⁺: 614

MS found (electrospray): (M+H)⁺ = 614

¹H NMR (CD₃OD) δ 8.25-8.20 (3H, m), 7.94 (2H, d), 7.54 (1 H, s), 7.03 (1 H, s), 6.44 (1 H, d), 4.78-4.67 (1 H, m), 4.01-3.86 (2H, m), 3.56-3.43 (2H, m), 2.23-0.93 (14H, m), carboxylic acid and amino protons not seen.

Example 28

S-IKfraπs^-MethylcyclohexylJcarbonyllttetrahydro^W-pyran^-ylmethylJaminol-S^- [1 ,3]oxazolo[4,5-jb]pyridin-2-ylphenyl)-2-thiophenecarboxylic acid

A mixture of Intermediate 44 (200 mg), Intermediate 60 (153 mg), sodium carbonate (168 mg) in water (2 ml.) and tetrakis(triphenylphosphine)palladium (0) (50 mg) in DMF (6 ml.) was stirred at 100⁰C overnight. The reaction was evaporated in vacuo and the residue was partitioned between DCM and saturated ammonium chloride. The organics were separated by passing through a hydrophobic frit and were evaporated in vacuo. The crude material was purified by reverse phase ISCO Companion chromatography, using a C18 cartridge, eluting with a gradient 40% MeCN (0.05% formic acid)/water (0.1% formic acid) to 95% MeCN (0.05% formic acid) to give the title compound. MS calcd for (C₃i H₃₃N₃O₅S)⁺: 560

MS found (electrospray): (M+H)⁺ = 560

¹H NMR (CD₃OD) δ 8.53 (1 H, d), 8.38 (2H, d), 8.15 (1 H, dd), 8.00 (2H, d), 7.69 (1 H, s), 7.48 (1 H, dd), 3.95-3.85 (3H, m), 3.43-3.31 (3H, m), 2.29-2.20 (1 H, m), 1.91-1.23 (12H, m), 0.79 (3H, d), 0.76-0.62 (2H, m), carboxylic acid proton not seen.

Example 29

S^^frans^-MethylcyclohexylJcarbonylJII -methyl^^methyloxyJethylJamino}^^- pyrazolo[1,5-a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylic acid

A solution of Intermediate 64 (73 mg) and 2M lithium hydroxide (1 ml.) in MeOH (4 ml.) was stirred at room temperature for 24 h, and was evaporated in vacuo. The residue was dissolved in water, acidified with 2M HCI, extracted with EtOAc, separated and dried using a hydrophobic frit. The crude material was purified by ISCO Companion silica chromatography, eluting with a gradient 0-100% MeOH in DCM. The material was purified further by MDAP HPLC and was freeze-dried from 1 ,4-dioxane to give the title compound. MS calcd for (C₂₉H₃₂N₄O₄S)⁺: 533 MS found (electrospray): (M+H)⁺ = 533

¹H NMR (de-DMSO) δ 13.50 (1 H, v.br.s), 9.17 (1 H, br.d), 8.57 (1 H, dd), 8.17 (2H, m), 7.92 (2H, m), 7.65, 7.50 (1 H, 2 x s), 7.35 (1 H, s), 7.07 (1 H, dd), 4.85, 4.60 (1 H, 2 x m), 3.57, 3.13 (3H, 2 x s), 3.40, 3.00 (2H, 2 x v.br.m), 2.03 (1 H, m), 1.70-1.40 (5H, m), 1.30-1.20 (2H, m), 1.15, 0.90 (3H, 2 x d), 0.73 (3H, d), 0.75-0.45 (2H, m), rotameric mixture observed. Example 30

5-(4-lmidazo[1,2-a]pyridin-2-ylphenyl)-3-(tetrahydro-2H-pyran-4-yl{[frans-4- (trifluoromethyl)cyclohexyl]carbonyl}amino)-2-thiophenecarboxylic acid

Intermediate 65 (70 mg) was dissolved in THF (0.3 ml.) and MeOH (0.3 ml_). 2N Sodium hydroxide solution (0.3 ml.) was added and the mixture was stirred at room temperature overnight. The reaction was evaporated in vacuo and was partitioned between DCM and 2M HCI. The aqueous was extracted with DCM and the combined organics were evaporated in vacuo to give the title compound. MS calcd for (C_3IH₃₀F₃N₃O₄S)⁺: 598 MS found (electrospray): (M+H)⁺ = 598

¹H NMR (CD₃OD) δ 8.81 (1 H, d), 8.66 (1 H, s), 8.06-7.89 (6H, m), 7.59 (1 H, s), 7.54-7.48 (1 H, m), 4.80-4.66 (1 H, m), 4.01-3.86 (2H, m), 3.56-3.43 (2H, m), 2.23-0.90 (14H, m), carboxylic acid proton not seen.

Example 31

3-{(frans-4-Hydroxycyclohexyl)[(frans-4-methylcyclohexyl)carbonyl]amino}-5-(4- pyrazolo[1,5-a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylic acid Example 31 was prepared as disclosed in Scheme 1 below.

MS calcd for (C_3IH₃₄N₄O₄S+ H): 559 MS found (electrospray): (M+H)⁺ = 559

Example 31

Scheme 1

Compound A

S-^-MethylcyclohexanecarbonylJ^etrahydropyran^-ylJaminol-S-phenyl-thiophene^- carboxylic acid

Intermediate 30 (320 mg) was dissolved in THF (5 mL) and ethanol (5 ml_). 2 M Lithium Hydroxide solution (5 mL) was then added. The reaction was stirred at room temperature for 16 h, then quenched with 2M hydrochloric acid (5 mL), and partitioned between water and dichloromethane. The layers were stirred and then separated using a hydrophobic frit, and the organic phase concentrated by evaporation. The residue was then loaded onto a 10g aminopropyl SPE cartridge in minimal 1 ,4-dioxane, washed with 4 column volumes of 1 ,4- dioxane, and then the product eluted using 10% acetic in 1 ,4-dioxane. The product fractions were then combined and concentrated by evaporation to give a residue, which was then further purified by HPLC using a Supelco ABZ+ plus 150 x 30 mm column, eluting with a gradient of 45-99% acetonitrile/ formic acid in water, over 15 minutes, at a flow rate of 40 mL/min, to give the title compound. MS calcd for (C₂₄H₂₉NO₃S + H)⁺: 428 MS found (electrospray): (M+H)⁺ = 428 1H NMR (CDCI₃): δ 6.68 (2H, d), 7.88 (1 H, d), 7.51- 7.44 (3H, m), 7.08 (1 H, s), 4.87 (1 H, bt), 3.96 (2H, dd), 3.50 (2H, dt), 2.07 (1 H, bt), 1.92 (1 H, d), 1.77 (1 H, d), 1.73-1.57 (6H, bm), 1.52-1.28 (3H, m), 0.79 (3H, d), 0.77-0.58 (2H, m), carboxylic acid proton not seen.

Compound B 3-[(2-Methoxy-1 -methyl-ethyl)(4-methylcyclohexanecarbonyl)amino]-5-phenyl- thiophene-2-carboxylic acid

To Intermediate 32 (56 mg; 0.11 mmol) in ethanol (5 ml.) was added 2M lithium hydroxide (0.55 ml_; 1.1 mmol). The reaction was stirred at room temperature for 16 hours and then evaporated to dryness. The residue was acidified with 2M HCI and purified by Oasis cartridge eluted with water and MeOH, and then purified by 5 g silica SPE cartridge eluted with increasing polarities of cyclohexane to DCM to EtOAc to give the title compound as an off white solid. MS calcd for (C₂₃H₂₉NO₄S+ H)⁺: 416 MS found (electrospray): (M+H)⁺ = 416

¹H NMR (DMSO): δ 7.81-7.75 (2H, m, 2 x CH [BOTH ROTAMERS]), 7.57 (¹/₃H, s, CH [MINOR ROT]), 7.52-7.41 (3H, m, 3 x CH [BOTH ROTS]), 7.40 (²/₃H, s, CH [MAJOR ROT]), 4.85 (²/₃H, m, CH [MAJOR ROT]), 4.58 (¹/₃H, m, CH [MINOR ROT]), 3.38 (¹/₃H, m, CH [MINOR ROT]), 3.30 (²/₃ x 2 CH, d, CH₂ [MAJOR ROT]), 3.27 (²/₃ x 3H, s, CH₃ [MAJOR ROT], 3.11 (¹/₃H + ¹/₃ χ 3H, m+s, CH + CH₃ [MINOR ROT]), 2.05 (1 H, m, CH [BOTH ROTS]), 1.68- 1.18 (7H, 2 x m, 7 x CH [BOTH ROTS], 1.15 (¹/₃ χ 3H, d, CH₃ [MINOR ROT]), 0.88 (²/₃ x 3H, d, CH₃ [MAJOR ROT]), 0.74 (3H, d, CH₃) [BOTH ROTS]), 0.70-0.45 (2H, 2 x m, 2 x CH [BOTH ROTS]).

The compounds according to the invention may be formulated for administration in any convenient way, and the invention therefore also includes within its scope pharmaceutical compositions for use in therapy, comprising a compound of Formula (I) or a pharmaceutically acceptable salt or solvate thereof in admixture with one or more pharmaceutically acceptable diluents or carriers. The compounds of the present invention can be administered by different routes including intravenous, intraperitoneal, subcutaneous, intramuscular, oral, topical, transdermal, or transmucosal administration. For systemic administration, oral administration is preferred. For oral administration, for example, the compounds can be formulated into conventional oral dosage forms such as capsules, tablets and liquid preparations such as syrups, elixirs and concentrated drops.

Alternatively, injection (parenteral administration) may be used, e.g., intramuscular, intravenous, intraperitoneal, and subcutaneous. For injection, the compounds of the invention are formulated in liquid solutions, preferably, in pharmaceutically compatible buffers or solutions, such as saline solution, Hank's solution, or Ringer's solution. In addition, the compounds may be formulated in solid form and redissolved or suspended immediately prior to use. Lyophilized forms can also be produced.

Systemic administration can also be by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art, and include, for example, for transmucosal administration, bile salts and fusidic acid derivatives. In addition, detergents may be used to facilitate permeation. Transmucosal administration, for example, may be through nasal sprays, rectal suppositories, or vaginal suppositories.

For topical administration, the compounds of the invention can be formulated into ointments, salves, gels, or creams, as is generally known in the art.

The amounts of various compounds to be administered can be determined by standard procedures taking into account factors such as the compound (IC₅₀) potency, (EC₅₀) efficacy, and the biological half-life (of the compound), the age, size and weight of the patient, and the disease or disorder associated with the patient. The importance of these and other factors to be considered are known to those of ordinary skill in the art.

Amounts administered also depend on the routes of administration and the degree of oral bioavailability. For example, for compounds with low oral bioavailability, relatively higher doses will have to be administered. Oral administration is a preferred method of administration of the present compounds.

Preferably the composition is in unit dosage form. For oral application, for example, a tablet, or capsule may be administered, for nasal application, a metered aerosol dose may be administered, for transdermal application, a topical formulation or patch may be administered and for transmucosal delivery, a buccal patch may be administered. In each case, dosing is such that the patient may administer a single dose. Each dosage unit for oral administration contains suitably from 0.01 to 500 mg/Kg, and preferably from 0.1 to 50 mg/Kg, of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, calculated as the free base. The daily dosage for parenteral, nasal, oral inhalation, transmucosal or transdermal routes contains suitably from 0.01 mg to 100 mg/Kg, of a compound of Formula(l). A topical formulation contains suitably 0.01 to 5.0% of a compound of Formula (I). The active ingredient may be administered from 1 to 6 times per day, preferably once, sufficient to exhibit the desired activity, as is readily apparent to one skilled in the art.

Compounds of Formula (I) which are active when given orally can be formulated as syrups, tablets, capsules and lozenges. A syrup formulation will generally consist of a suspension or solution of the compound or salt in a liquid carrier for example, ethanol, peanut oil. olive oil, glycerine or water with a flavouring or colouring agent. Where the composition is in the form of a tablet, any pharmaceutical carrier routinely used for preparing solid formulations may be used. Examples of such carriers include magnesium stearate, terra alba, talc, gelatin, acacia, stearic acid, starch, lactose and sucrose. Where the composition is in the form of a capsule, any routine encapsulation is suitable, for example using the aforementioned carriers in a hard gelatin capsule shell. Where the composition is in the form of a soft gelatin shell capsule any pharmaceutical carrier routinely used for preparing dispersions or suspensions may be considered, for example aqueous gums, celluloses, silicates or oils, and are incorporated in a soft gelatin capsule shell.

Typical parenteral compositions consist of a solution or suspension of a compound or salt in a sterile aqueous or non-aqueous carrier optionally containing a parenterally acceptable oil, for example polyethylene glycol, polyvinylpyrrolidone, lecithin, arachis oil or sesame oil.

Typical compositions for inhalation are in the form of a solution, suspension or emulsion that may be administered as a dry powder or in the form of an aerosol using a conventional non- CFC propellant such as 1 ,1 ,1 ,2-tetrafluoroethane or 1 ,1 ,1 ,2,3,3,3-heptafluoropropane.

A typical suppository formulation comprises a compound of Formula (I) or a pharmaceutically acceptable salt thereof which is active when administered in this way, with a binding and/or lubricating agent, for example polymeric glycols, gelatins, cocoa-butter or other low melting vegetable waxes or fats or their synthetic analogs.

Typical dermal and transdermal formulations comprise a conventional aqueous or nonaqueous vehicle, for example a cream, ointment, lotion or paste or are in the form of a medicated plaster, patch or membrane.

ASSAYS

The potential for compounds of the invention to inhibit NS5B wildtype HCV polymerase activity, genotype 1 b, may be demonstrated, for example, using the following in vitro assay: In Vitro Detection of inhibitors of HCV RNA-dependent RNA Polymerase Activity

Incorporation of [³³P]-GMP into RNA was followed by absorption of the biotin labelled RNA polymer by streptavidin containing SPA beads. A synthetic template consisting of biotinylated 13mer-oligoG hybridised to polyrC was used as a homopolymer substrate.

Genotype 1b Full-Length Enzyme

Reaction Conditions were 0.5 μM [³³P]-GTP (20 Ci/mMol), 1 mM Dithiothreitol, 20 mM MgCI₂, 5mM MnCI₂, 20 mM Tris-HCI, pH7.5, 1.6 μg/mL polyC/0.256 μM biotinylated oligoG13, 10% glycerol, 0.01% NP-40, 0.2 u/μL RNasin and 50 mM NaCI.

HCV RNA Polymerase (Recombinant full-length NS5B (Lohmann et al, J. Virol. 71 (1 1 ), 1997, 8416. 'Biochemical properties of hepatitis C virus NS5B RNA-dependent RNA polymerase and identification of amino acid sequence motifs essential for enzymatic activity') expressed in baculovirus and purified to homogeneity) was added to 4 nM final concentration.

5x concentrated assay buffer mix was prepared using 1 M MnCI₂ (0.25 ml_), glycerol (2.5ml_), 10% NP-40 (0.025 ml.) and Water (7.225 ml_), Total 10 ml_.

2x concentrated enzyme buffer contained 1 M-Tris-HCI, pH7.5 (0.4 ml_), 5M NaCI (0.2 ml_), 1 M-MgCI₂ (0.4 ml_), glycerol (1 ml_), 10% NP-40 (10 μl_), 1 M DTT (20 μl_) and water (7.97 ml_), Totaπ O ml_.

Substrate Mix was prepared using 5x Concentrated assay Buffer mix (4μl_), [³³P]-GTP (10 μCi/μL, 0.02μl_), 25 μM GTP (0.4 μl_), 40 u/μL RNasin (0.1 μl_), 20 μg/mL polyrC/biotinylated- oligorG (1.6 μl_), and Water (3.94 μl_), Total 10 μl_.

Enzyme Mix was prepared by adding 1 mg/ml full-length NS5B polymerase (1.5 μl_) to 2.81 ml. 2x-concentrated enzyme buffer.

The Assay was set up using compound (1 μL), Substrate Mix (10 μl_), and Enzyme Mix (added last to start reaction) (10 μl_), Tote/ 21 μl_.

The reaction was performed in a U-bottomed, white, 96-well plate. The reaction was mixed on a plate-shaker, after addition of the Enzyme, and incubated for 1 h at 22⁰C. After this time, the reaction was stopped by addition of 40 μl_ 1.875 mg/ml streptavidin SPA beads in 0.1 M EDTA. The beads were incubated with the reaction mixture for 1 h at 22⁰C after which 120 μl_ 0.1 M EDTA in PBS was added. The plate was sealed, mixed centrifuged and incorporated radioactivity determined by counting in a Trilux (Wallac) or Topcount (Packard) Scintillation Counter. After subtraction of background levels without enzyme, any reduction in the amount of radioactivity incorporated in the presence of a compound, compared to that in the absence, was taken as a measure of the level of inhibition. Ten concentrations of compounds were tested in three- or fivefold dilutions. From the counts, percentage of inhibition at highest concentration tested or IC₅₀S for the compounds were calculated using GraFit 3, GraFit 4 or GraFit 5 software packages or a data evaluation macro for Excel based on XLFit Software (IDBS).

The potential for compounds of the invention to inhibit HCV replication, genotype 1 a and genotype 1 b, may be demonstrated, for example, using the following cell based assay:

Replicon ELISA cell based assay

Method

100 μl_ of medium containing 10% FCS were added to each well of clear, flat-bottomed 96 well microplates, excepting wells in the top row. Test compound was diluted in assay medium to twice the final required starting concentration from a 40 mM stock solution in DMSO. 200 μl_ of the starting dilution were introduced into two wells each in the top row and doubling dilutions made down the plate by the sequential transfer of 100 μl_ aliquots with thorough mixing in the wells; the final 100 μl_ were discarded. The two bottom rows were not used for compound dilutions. Huh-7 HCV replicon cell monolayers nearing confluency were stripped from growth flasks with versene-trypsin solution and the cells were resuspended in assay medium at either 2 x 10⁵ cells/mL (sub-line 5-15; genotype 1 b; Lohmann, V., Korner, F., Koch, J-O., Herian, U., Thielmann, L. and Bartenschlager, R., Science, 1999, 285, 110- 1 13) or at 3 x 10⁵ cells/mL (genotype 1 a; Gu, B., Gates, AT., Isken, O., Behrens, S. E. and Sarisky, R.T., J. Virol., 2003, 77, 5352-5359). 100 μl_ of cell suspension were added to all wells and the plates incubated at 37°C for 72 hours in a 5% CO₂ atmosphere. Following incubation, the assay medium was aspirated from the plates. The cell sheets were washed by gentle immersion in phosphate buffered saline (PBS), which was then aspirated off, and fixed with acetone:methanol (1 :1 ) for 5 minutes. Following a further wash with PBS, 100 μL of ELISA diluent (PBS + 0.05% v/v Tween 20 + 2% w/v skimmed milk powder) were added to all wells and the plates incubated at 37°C for 30 minutes on an orbital platform. The diluent was removed and each well then received 50 μL of a 1/200 dilution of anti-HCV specific, murine, monoclonal antibody (either Virostat #1872 or #1877), except for wells in one of the compound-free control rows which received diluent alone to act as negative controls. The plates were incubated at 37°C for 2 hours and washed 3 times with PBS/0.05% Tween 20, then 50 μL of horseradish peroxidase conjugated, anti-mouse, rabbit polyclonal serum (Dako #P0260), diluted 1/1000, were added to all wells. The plates were incubated for a further hour, the antibody removed and the cell sheets washed 5 times with PBS/Tween and blotted dry. The assay was developed by the addition of 50 μL of ortho- phenylenediamine/peroxidase substrate in urea/citrate buffer (SigmaFast, Sigma #P-9187) to each well, and colour allowed to develop for up to 15 minutes. The reaction was stopped by the addition of 25 μl_ per well of 2 M sulphuric acid and the plates were read at 490 nm on a Fluostar Optima spectrophotometer.

The substrate solution was removed and the plates were washed in tap water, blotted dry and the cells stained with 5 % carbol fuchsin in water for 30 minutes. The stain was discarded and the cell sheets washed, dried and examined microscopically to assess cytotoxicity. Data analysis

The absorbance values from all compound-free wells that had received both primary and secondary antibodies were averaged to obtain a positive control value. The mean absorbance value from the compound-free wells that had not received the primary antibody was used to provide the negative (background) control value. The readings from the duplicate wells at each compound concentration were averaged and, after the subtraction of the mean background from all values, were expressed as a percentage of the positive control signal. The quantifiable and specific reduction of expressed protein detected by the ELISA in the presence of a drug can be used as a measure of replicon inhibition. GraFit software (Erithacus Software Ltd.) was used to plot the curve of percentage inhibition against compound concentration and derive the 50% inhibitory concentration (IC₅₀) for the compound.

Replicon Luciferase cell based assay

Method

A 4OmM stock solution in DMSO of each test compound was further diluted into 50μL of DMSO in the first row of a 96 well, V-bottom microplate, to give 100 times the top concentration of the required dilution series. Aliquots of 25μL of DMSO were added to each well of the remaining rows, and doubling dilutions of compound were prepared by the serial transfer of 25μL volumes from the first row onwards. A Plate-mate robot was used to transfer 1 μL volumes from each dilution well into duplicate wells of a clear bottom, black walled, 96 well assay plate (COSTAR #3603). Control wells received 1 μL of DMSO alone.

Suspensions were prepared from cultures of Huh-7 cells stably transfected with sub-genomic HCV NS3-NS5B replicons of either genotype 1 b (the ET subline described by Pietschmann,T., Lohmann, V., Kaul, A., Krieger, N., Rinck, G., Rutter, G., Strand, D. & Bartenschlager, R., Journal of Virology, 2002, 76, 4008-4021 ) or genotype 1a (subline 1.19 constructed in-house) linked to a firefly luciferase reporter gene. Monolayers nearing confluency were stripped from growth flasks with versene-trypsin solution and the cells re- suspended in assay medium comprising DMEM (Invitrogen #41965-039) supplemented with 5% v/v foetal calf serum, 1% v/v non-essential amino acids solution, 100 units/ml penicillin, 100μg/ml streptomycin and 2mM L-glutamine. 100μL of suspension containing either 15,000 cells (genotype 1 b luciferase replicon) or 20,000 cells (genotype 1a luciferase replicon) were added to all wells, except medium controls, of the assay plate and the plate incubated for 48 hours at 37°C in a 5% CO₂ atmosphere. One tablet of Resazurin (Fisher #R/0040/79) was dissolved in 5OmL of phosphate buffered saline and 100μl_ of solution added to all wells. The plate was re-incubated at 37°C for a further 2- 4 hours, wrapped in aluminium foil, before reading in a FluoStar Optima at 595nm. All growth medium and Resazurin was removed by aspiration, and an opaque mask applied to the bottom of the plate. A solution of SteadyLite cytolytic buffer/I uciferase substrate (Perkin-Elmer #6016987) was prepared according to the manufacturer's instructions, and 25μl_ added to each well. The plate was then read for luminescence on a TopCount NXT.

Data Analysis Toxicity : The Resazurin absorbance values from duplicate wells were averaged and expressed as a percentage of the mean absorbance of compound free control wells to determine comparative cell viability. Compound cytotoxicity was expressed either as the lowest concentration at which a significant reduction in viability was observed or a 50% toxic concentration (CCID₅₀) was determined by plotting percentage cytotoxicity against compound concentration using Grafit software (Erithacus Software Ltd.).

Potency : The luminescence values from all compound-free wells containing cells were averaged to obtain a positive control value. The mean luminescence value from the compound-free wells that had received no cells was used to provide the negative (background) control value. The readings from the duplicate wells at each compound concentration were averaged and, after the subtraction of the mean background from all values, were expressed as a percentage of the positive control signal. The quantifiable and specific reduction of luciferase signal in the presence of a drug is a direct measure of replicon inhibition. GraFit software was used to plot the curve of percentage inhibition against compound concentration and derive the 50% inhibitory concentration (IC₅₀) for the compound.

Results

All compounds were assayed at least twice and the mean data are included in the table below.

^* For these example compounds, the procedure described above was followed but the test compound was introduced into a single well in the top row of the microplate only, with the result that the compound was assayed only once and the data shown are thus from a single experiment.

Activity ranges

Genotype 1a Genotype 1 b

+ ≥0.5000 μM + ≥0.5000 μM

++ 0.10 - 0.4900 μM ++ 0.10 - 0.4900 μM

+++ 0.05 - 0.0990 μM +++ 0.05 - 0.0990 μM

++++ 0.01 - 0.0490 μM ++++ 0.01 - 0.0490 μM

+++++ 0.005 - 0.0099 μM +++++ 0.005 - 0.0099 μM

++++++ 0.001 - 0.0049 μM ++++++ 0.001 - 0.0049 μM

+++++++ <0.001 μM +++++++ <0.001 μM

Compound A corresponds to the compound disclosed as Example 572 in WO2002/100851 ,

3-[(4-methylcyclohexanecarbonyl)(tetrahydropyran-4-yl)amino]-5-phenyl-thiophene-2- carboxylic acid.

Compound B corresponds to the compound disclosed as Example 578 in WO2002/100851 ,

3-[(2-methoxy-1-methyl-ethyl)(4-methylcyclohexanecarbonyl)amino]-5-phenyl-thiophene-2- carboxylic acid.

Compounds A to B may be made according to the processes described in WO2002/100851 or as described hereinabove.

Structures of Compounds A-B are shown below for the avoidance of doubt.

The compounds of the present invention which have been tested demonstrate a surprisingly superior potency as HCV polymerase inhibitors, as shown by the IC₅O values in the cell- based assays across both of the 1a and 1 b genotypes of HCV, compared to Compounds A - B. Accordingly, the compounds of the present invention are of great potential therapeutic benefit in the treatment and prophylaxis of HCV.

The pharmaceutical compositions according to the invention may also be used in combination with other therapeutic agents, for example immune therapies [e.g. Interferon, such as Interferon alfa-2a (Roferon-A; Hoffmann-La Roche), interferon alpha-2b (Intron-A; Schering-Plough), interferon alfacon-1 (Infergen; Intermune), peginterferon alpha-2b (Peg- Intron; Schering-Plough) or peginterferon alpha-2a (Pegasys; Hoffmann-La Roche)], therapeutic vaccines, antifibrotic agents, anti-inflammatory agents [such as corticosteroids or NSAIDs], bronchodilators [such as beta-2 adrenergic agonists and xanthines (e.g. theophylline)], mucolytic agents, anti-muscarinics, anti-leukotrienes, inhibitors of cell adhesion [e.g. ICAM antagonists], anti-oxidants [e.g. N-acetylcysteine], cytokine agonists, cytokine antagonists, lung surfactants and/or antimicrobial, anti-viral agents [e.g. ribavirin and amantidine], and anti-HCV agents [for example HCV NS3 protease inhibitors, e.g. VX950 (telapravir; Vertex) or SCH503034 (Schering Plough)], or HCV NS5b polymerase inhibitors [for example HCV796 (Wyeth) or R1626 (Roche)], RNAi agents or cyclophilin inhibitors. The compositions according to the invention may also be used in combination with gene replacement therapy.

The invention thus provides, in a further aspect, a combination comprising a compound of Formula (I) together with at least one other therapeutically active agent, especially Interferon, ribavirin and/or an additional anti-HCV agent.

The combinations referred to above may conveniently be presented for use in the form of a pharmaceutical formulation and thus pharmaceutical formulations comprising a combination as defined above together with a pharmaceutically acceptable carrier thereof represent a further aspect of the invention.

The individual components of such combinations may be administered either sequentially or simultaneously in separate or combined pharmaceutical formulations. Appropriate doses of known therapeutic agents will be readily appreciated by those skilled in the art.

All publications, including but not limited to patents and patent applications cited in this specification are herein incorporated by reference as if each individual publication were specifically and individually indicated to be incorporated by reference as though fully set forth.

Claims

Claims

A compound of Formula (I):

wherein: A represents hydroxy;

R¹ represents -R^x-R^γ;

R^x represents phenyl (optionally substituted at one of the mefø-positions to the thiophene by halo, methyl, ethyl or trifluoromethyl) or a 5- or 6-membered heteroaryl (optionally substituted at one of the mefø-positions to the thiophene by halo, methyl, ethyl, methoxy or trifluoromethyl) bonded through a ring carbon atom to the carbon atom of the thiophene;

R^γ represents a 8, 9 or 10-membered heteroaryl, bonded such that when R^x is phenyl or a 6- membered heteroaryl, R^γ is in the para-position;

R² represents C_5-7cycloalkyl optionally substituted by one or more substituents selected from fluoro, =CHCH₃, OH, OCH₃, =0 or -Ci_-6alkyl (wherein the alkyl may itself be optionally substituted by one or more fluoro atoms); with the proviso that when R¹ represents 4-imidazo[2,1-b][1 ,3]thiazol-6-ylphenyl, then R² may not represent trans-4- (trifluoromethyl)cyclohexyl;

R³ represents -(CH₂)₂-O-(CH₂)_mH, -CH-(CH₂OMe)₂, -CH(R⁴)-CH₂-O-(CH₂)_mH, - CH₂-CH(R⁴)- O-(CH₂)_mH, tetrahydrofuran-3-yl, -CH₂-tetrahydrofuran-3-yl, tetrahydro-2/-/-pyran-4-yl, -CH₂- tetrahydro-2/-/-pyran-4-yl or frans-4-hydroxycyclohex-1-yl;

R⁴ represents Ci_-3alkyl

m represents an integer selected from 1 or 2;

or salts, solvates or esters thereof

2. A compound of Formula (I) selected from the group consisting of: 3-[[(trans-4-methylcyclohexyl)carbonyl](tetrahydro-3-furanyl)amino]-5-(4-pyrazolo[1 ,5- a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylic acid;

3-[[(trans-4-methylcyclohexyl)carbonyl](tetrahydro-2H-pyran-4-yl)amino]-5-(4-pyrazolo[1 ,5- a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylic acid; 5-(4-furo[3,2-b]pyridin-2-ylphenyl)-3-[[(trans-4-methylcyclohexyl)carbonyl](tetrahydro-2H- pyran-4-yl)amino]-2-thiophenecarboxylic acid;

5-[4-(7-aminopyrazolo[1 ,5-a]pyrimidin-2-yl)phenyl]-3-[[(trans-4- methylcyclohexyl)carbonyl](tetrahydro-2H-pyran-4-yl)amino]-2-thiophenecarboxylic acid; 5-(4-imidazo[1 ,2-a]pyridin-2-ylphenyl)-3-[[(trans-4-methylcyclohexyl)carbonyl](tetrahydro-2H- pyran-4-yl)amino]-2-thiophenecarboxylic acid;

5-(4-ilmidazo[2,1-b][1 ,3]thiazol-6-ylphenyl)-3-[[(trans-4- methylcyclohexyl)carbonyl](tetrahydro-2H-pyran-4-yl)amino]-2-thiophenecarboxylic acid;

5-(4-furo[3,2-b]pyridin-2-ylphenyl)-3-{[(trans-4-methylcyclohexyl)carbonyl][2- (methyloxy)ethyl]amino}-2-thiophenecarboxylic acid;

5-[4-(7-aminopyrazolo[1 ,5-a]pyrimidin-2-yl)phenyl]-3-{[(trans-4-methylcyclohexyl)carbonyl][2-

(methyloxy)ethyl]amino}-2-thiophenecarboxylic acid;

3-{[(trans-4-methylcyclohexyl)carbonyl][2-(methyloxy)ethyl]amino}-5-(4-pyrazolo[1 ,5- alpyrimidin^-ylphenyl^-thiophenecarboxylic acid; 5-(4-imidazo[1 ^-alpyridin^-ylpheny^-S-^trans^-methylcyclohexyOcarbonyl]!^--

(methyloxy)ethyl]amino}-2-thiophenecarboxylic acid;

5-(4-imidazo[2,1-b][1 ,3]thiazol-6-ylphenyl)-3-{[(trans-4-methylcyclohexyl)carbonyl][2-

(methyloxy)ethyl]amino}-2-thiophenecarboxylic acid;

3-{[2-(ethyloxy)ethyl][(trans-4-methylcyclohexyl)carbonyl]amino}-5-(4-furo[3,2-b]pyridin-2- ylphenyl)-2-thiophenecarboxylic acid;

5-[4-(7-aminopyrazolo[1 ,5-a]pyrimidin-2-yl)phenyl]-3-{[2-(ethyloxy)ethyl][(trans-4- methylcyclohexyl)carbonyl]amino}-2-thiophenecarboxylic acid;

3-{[2-(ethyloxy)ethyl][(trans-4-methylcyclohexyl)carbonyl]amino}-5-(4-pyrazolo[1 ,5- alpyrimidin^-ylphenyl^-thiophenecarboxylic acid; 3-{[2-(ethyloxy)ethyl][(trans-4-methylcyclohexyl)carbonyl]amino}-5-(4-imidazo[1 ,2-a]pyridin-2- ylphenyl)-2-thiophenecarboxylic acid;

3-{[2-(ethyloxy)ethyl][(trans-4-methylcyclohexyl)carbonyl]amino}-5-(4-imidazo[2,1- b][1 ,3]thiazol-6-ylphenyl)-2-thiophenecarboxylic acid;

3-([(trans-4-methylcyclohexyl)carbonyl]{2-(methyloxy)-1-[(methyloxy)methyl]ethyl}amino)-5- (4-pyrazolo[1 ,5-a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylic acid;

5-(4-furo[3,2-b]pyridin-2-ylphenyl)-3-[[(trans-4-methylcyclohexyl)carbonyl](tetrahydro-3- furanyl)amino]-2-thiophenecarboxylic acid;

5-(4-imidazo[1 ,2-a]pyridin-2-ylphenyl)-3-[[(trans-4-methylcyclohexyl)carbonyl](tetrahydro-2H- pyran-4-ylmethyl)amino]-2-thiophenecarboxylic acid; 5-(4-furo[3,2-b]pyridin-2-ylphenyl)-3-[[(trans-4-methylcyclohexyl)carbonyl](tetrahydro-2H- pyran-4-ylmethyl)amino]-2-thiophenecarboxylic acid; 3-[[(trans-4-methylcyclohexyl)carbonyl](tetrahydro-2H-pyran-4-ylmethyl)amino]-5-(4- pyrazolo[1 ,5-a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylic acid;

3-([2-(methyloxy)ethyl]{[trans-4-(trifluoromethyl)cyclohexyl]carbonyl}amino)-5-(4- pyrazoloπ .S-alpyrimidin^-ylphenyl^-thiophenecarboxylic acid; 5-(4-furo[3,2-b]pyridin-2-ylphenyl)-3-([2-(methyloxy)ethyl]{[trans-4-

(trifluoromethyl)cyclohexyl]carbonyl}amino)-2-thiophenecarboxylic acid;

5-(4-imidazo[2,1-b][1 ,3]thiazol-6-ylphenyl)-3-[[(trans-4-methylcyclohexyl)carbonyl](tetrahydro-

2H-pyran-4-ylmethyl)amino]-2-thiophenecarboxylic acid;

5-(4-pyrazolo[1 ,5-a]pyrimidin-2-ylphenyl)-3-(tetrahydro-2H-pyran-4-yl{[trans-4- (trifluoromethyl)cyclohexyl]carbonyl}amino)-2-thiophenecarboxylic acid;

5-(4-furo[3,2-b]pyridin-2-ylphenyl)-3-(tetrahydro-2H-pyran-4-yl{[trans-4-

(trifluoromethyl)cyclohexyl]carbonyl}amino)-2-thiophenecarboxylic acid formic acid salt;

5-[4-(7-aminopyrazolo[1 ,5-a]pyrimidin-2-yl)phenyl]-3-(tetrahydro-2H-pyran-4-yl{[trans-4-

(trifluoromethyl)cyclohexyl]carbonyl}amino)-2-thiophenecarboxylic acid; 3-[[(trans-4-methylcyclohexyl)carbonyl](tetrahydro-2H-pyran-4-ylmethyl)amino]-5-(4-

[i ^oxazolo^S-blpyridin^-ylphenyl^-thiophenecarboxylic acid;

3-{[(trans-4-methylcyclohexyl)carbonyl][1-methyl-2-(methyloxy)ethyl]amino}-5-(4- pyrazoloπ .δ-alpyrimidin^-ylphenyl^-thiophenecarboxylic acid;

5-(4-imidazo[1 ,2-a]pyridin-2-ylphenyl)-3-(tetrahydro-2H-pyran-4-yl{[trans-4- (trifluoromethyl)cyclohexyl]carbonyl}amino)-2-thiophenecarboxylic acid;

3-{(frans-4-Hydroxycyclohexyl)[(frans-4-methylcyclohexyl)carbonyl]amino}-5-(4-pyrazolo[1 ,5- a]pyrimidin-2-ylphenyl)-2-thiophenecarboxylic acid; and salts, solvates and esters, and individual enantiomers thereof where appropriate.

3. A compound as claimed in Claim 1 , wherein R^x represents phenyl optionally substituted by halo, methyl or trifluoromethyl.

4. A compound as claimed in Claim 1 or 3, wherein R^γ represents furo[3,2-fc>]pyridin-2-yl, pyrazolo[1 ,5-a]pyrimidin-2-yl, imidazo[1 ,2-a]pyridin-2-yl, imidazo[2,1-b][1 ,3]thiazol-6-yl, 7- amino-5-methylpyrazolo[1 ,5-a]pyrimidin-2-yl, 5-methylpyrazolo-[1 ,5-a]pyrimidin-2-yl, 7- aminopyrazolo[1 ,5-a]pyrimidin-2-yl, [1 ,3]oxazolo[4,5-b]pyridin-2-yl, furo[2,3-b]pyridin-5-yl, 5- amino-1 ,3-benzoxazol-2-yl, [1 ,3]oxazolo[5,4-b]pyridin-2-yl, or furo[3,2-c]pyridin-2-yl.

5. A compound as claimed in Claim 1 , 3 or 4, wherein R² represents trans-4- methylcyclohexyl.

6. A compound as claimed in Claim 1 , 3, 4 or 5, wherein R³ represents methoxyethyl, ethoxyethyl, tetrahydrofuran-3-yl, tetrahydro-2/-/-pyran-4-yl or frans-4-hydroxycyclohexyl.

7. A compound as claimed in Claim 1 wherein R^x represents phenyl optionally substituted by halo, methyl or trifluoromethyl; R^γ represents furo[3,2-t)]pyridin-2-yl, pyrazolo[1 ,5-a]pyrimidin-2-yl, imidazo[1 ,2-a]pyridin-2-yl, imidazo[2,1-ιb][1 ,3]thiazol-6-yl, 7- amino-5-methylpyrazolo[1 ,5-a]pyrimidin-2-yl, 5-methylpyrazolo-[1 ,5-a]pyrimidin-2-yl, 7- aminopyrazolo[1 ,5-a]pyrimidin-2-yl, [1 ,3]oxazolo[4,5-b]pyridin-2-yl, furo[2,3-b]pyridin-5-yl, 5- amino-1 ,3-benzoxazol-2-yl, [1 ,3]oxazolo[5,4-b]pyridin-2-yl, or furo[3,2-c]pyridin-2-yl; R² represents C₆cycloalkyl optionally substituted by one or more Ci_-4alkyl substituents (wherein the alkyl group may itself be optionally substituted by one or more fluoro atoms); and R³ represents -(CHz)₂-O-(CH₂VH, -CH-(CH₂OMe)₂, -CH(Me)CH₂-O-Me, tetrahydrofuran-3-yl, tetrahydro-2/-/-pyran-4-yl, -CH₂-tetrahydro-2/-/-pyran-4-yl or frans-4-hydroxycyclohexyl.

8. A compound as claimed in Claim 1 or Claim 7 wherein R^x represents unsubstituted phenyl; R^γ represents furo[3,2-b]pyridin-2-yl, pyrazolo[1 ,5-a]pyrimidin-2-yl, imidazo[1 ,2- a]pyridin-2-yl, imidazo[2,1-b][1 ,3]thiazol-6-yl, 7-aminopyrazolo[1 ,5-a]pyrimidin-2-yl or [1 ,3]oxazolo[4,5-b]pyridin-2-yl; R² represents frans-4-methylcyclohexyl or trans-4- (trifluoromethyl)cyclohexyl; and R³ represents methoxyethyl, ethoxyethyl, 1-methyl-2- (methyloxy)ethyl, tetrahydrofuran-3-yl, tetrahydro-2/-/-pyran-4-yl or trans-4- hydroxycyclohexyl.

9. A method of treating or preventing viral infection which comprises administering to a subject in need thereof, an effective amount of a compound of Formula (I):

wherein:

A represents hydroxy;

R¹ represents -R^x-R^γ;

R^x represents phenyl (optionally substituted at one of the mefø-positions to the thiophene by halo, methyl, ethyl or trifluoromethyl) or a 5- or 6-membered heteroaryl (optionally substituted at one of the mefø-positions to the thiophene by halo, methyl, ethyl, methoxy or trifluoromethyl) bonded through a ring carbon atom to the carbon atom of the thiophene;

R^γ represents a 8, 9 or 10-membered heteroaryl, bonded such that when R^x is phenyl or a 6- membered heteroaryl, R^γ is in the para-position;

R² represents C_5-7cycloalkyl optionally substituted by one or more substituents selected from fluoro, =CHCH₃, OH, OCH₃, =0 or -Ci_-6alkyl (wherein the alkyl may itself be optionally substituted by one or more fluoro atoms); with the proviso that when R¹ represents 4-imidazo[2,1-b][1 ,3]thiazol-6-ylphenyl, then R² may not represent trans-4- (trifluoromethyl)cyclohexyl;

R³ represents -(CHz)₂-O-(CH₂VH, -CH-(CH₂OMe)₂, -CH(R⁴)-CH₂-O-(CH₂)_mH, - CH₂-CH(R⁴)- O-(CH₂)_mH, tetrahydrofuran-3-yl, -CH₂-tetrahydrofuran-3-yl, tetrahydro-2/-/-pyran-4-yl, -CH₂- tetrahydro-2/-/-pyran-4-yl or frans-4-hydroxycyclohex-1-yl;

R⁴ represents Ci_-3alkyl

m represents an integer selected from 1 or 2;

and salts, solvates and esters thereof

10. A method as claimed in claim 9 which involves inhibiting HCV replication.

1 1. A compound of Formula (I) :

wherein: A represents hydroxy;

R¹ represents -R^x-R^γ;

R^x represents phenyl (optionally substituted at one of the mefø-positions to the thiophene by halo, methyl, ethyl or trifluoromethyl) or a 5- or 6-membered heteroaryl (optionally substituted at one of the mefø-positions to the thiophene by halo, methyl, ethyl, methoxy or trifluoromethyl) bonded through a ring carbon atom to the carbon atom of the thiophene;

R^γ represents a 8, 9 or 10-membered heteroaryl, bonded such that when R^x is phenyl or a 6- membered heteroaryl, R^γ is in the para-position;

R² represents C_5-7cycloalkyl optionally substituted by one or more substituents selected from fluoro, =CHCH₃, OH, OCH₃, =0 or -Ci_-6alkyl (wherein the alkyl may itself be optionally substituted by one or more fluoro atoms); with the proviso that when R¹ represents 4-imidazo[2,1-b][1 ,3]thiazol-6-ylphenyl, then R² may not represent trans-4- (trifluoromethyl)cyclohexyl; R³ represents -(CHz)₂-O-(CH₂VH, -CH-(CH₂OMe)₂, -CH(R⁴)-CH₂-O-(CH₂)_mH, - CH₂-CH(R⁴)- O-(CH₂)_mH, tetrahydrofuran-3-yl, -CH₂-tetrahydrofuran-3-yl, tetrahydro-2/-/-pyran-4-yl, -CH₂- tetrahydro-2/-/-pyran-4-yl or frans-4-hydroxycyclohex-1-yl;

R⁴ represents Ci_-3alkyl

m represents an integer selected from 1 or 2;

and salts, solvates and esters thereof

for use in medical therapy.

12. A compound as claimed in claim 1 1 wherein the medical therapy is the treatment of viral infection.

13. A compound as claimed in claim 12 wherein the viral infection is HCV.

14. A pharmaceutical formulation comprising a compound of Formula (I) or pharmaceutically acceptable salts, solvates or esters thereof as defined in claim 1 in conjunction with at least one pharmaceutically acceptable diluent or carrier.

15. A pharmaceutical formulation as claimed in claim 14, presented in an oral dosage form.

16. A process for the preparation of a compound of Formula (I) as defined in claim 1 , comprising deprotection of a

in which A is an alkoxy y,, bbeennzzyylloxy or silyloxy group, and R¹, R² and R³ are as defined for

Formula (I) in Claim 1.

17. Use of a compound of Formula (I) or pharmaceutically acceptable salts, solvates or esters thereof as defined in claim 1 , in the manufacture of a medicament for the treatment and/or prophylaxis of viral infection.

18. Use as claimed in claim 17 wherein the viral infection is HCV.

19. A combination comprising a compound of Formula (I) as defined in Claim 1 , together with at least one other therapeutically active agent.

20. A combination as claimed in Claim 19, wherein the other therapeutically active agent is selected from Interferon, ribavirin and/or an additional anti-HCV agent.