WO2005066156A1 - Selective kinase inhibitors - Google Patents

Abstract

A compound of the general formula (I) or pharmaceutically acceptable prodrugs, salts, hydrates, solvates, crystal forms or diastereomers thereof, wherein A represents a variety of six membered nitrogen containing heterocyclic rings, Q is a bond, halogen, C1-4 alkyl, O, S, SO2, CO or CS and X1, X2, X3 and X4 are optionally substituted by 9 specific substituents or one can be nitrogen. Compositions comprising a carrier and at least one compound of formula (I) are also provided. Further provided are methods of treating tyrosine kinase-associated disease states by administering a compound of formula (I) and methods of suppressing the immune system of a subject by administering a compound of formula (I).

Description

I SELECTIVE KINASE INHIBITORS FIELD OF THE INVENTION

The present invention relates to the field of inhibitors of protein tyrosine kinases in particular the JAK family of protein tyrosine kinases.

BACKGROUND OF THE INVENTION Protein kinases are a family of enzymes that catalyse the phosphoryϊation of specific residues in proteins. In general protein kinases fall into several groups; those which preferentially phosphoiylate serine and/ or threonine residues, those which preferentially phosphorylate tyrosine residues and those which phosphorylate both tyrosine and Ser/Thr residues. Protein kinases are therefore key elements in signal transduction pathways responsible for transducing extracellular signals, incl uding the action of cytokines on their receptors, to the nuclei, triggering various biological events. The many roles of protein kinases in normal cell physiology include cell c cle control and cell growth, differentiation, apoptosis, cell mobility and mitogenesis.

Frotein kinases include, for example, but arc not limited to, members of the Protein Tyrosine Kinase family (PTKs), which in turn can be divided into the cy toplasmic PTKs and the receptor PTKs (RTKs). The cytoplasmic PTKS include the SRC family, (mduding: BLK; FGR; FYN; HCK; T.CK; LYN; SR YES and YKK); the BRK Family (including: BRK; FRK, SAD; and SKM); the CSK f mily (including: CS and CTK); the BTK family, (including BTK; TTK; TEC; KK2 and TXK), the Janus kinase family, (including: JAKT, JAK2, JAK3 and Tyk2), the FA family (including, FAK and PYK2); the Fes family (including FES and FER), the ZAP70 family (including ZAP70 and SYK); the ACK family (including ACK1 and AC 2); and the Abl family (including ABL and ARG). The RTK f-unily includes the EGF-Receptor family (including, EG R, HER2_/ HER3 and HER4); the Insulin Receptor family (including INS-R and IGF1-R ); the PDGF-Receptor family (including PDGFRoe, FDGFRβ, CSF'IR, KIT, FLK2 ); the VEGF-Receptor family (including; FLT1, FLK1 and FLT4); the FGF-Reccptor family (including FGFR1, FCFR2, FGFR3 and FGFR4 ); the CCK4 family (including CCK4); the MET family (including MET and RON); the TRK family (including TRKA, TRKB, and TRKC ); the AXL family (including AXL, MER, and SKY); the TTE/TEK family (including TIE and TTE2/TEK); the EFH family (including EFHAl, EPHA2, ETΗA3, EFHΛ4, EPHA5, EPHA6, EPHA7, EPHA8, EPHB1, EPHB2, EPHB3, EPHB4, EPHB5, EPHB6); the RYK family (including RYK); the MCK family (including MCK and TYRO10); the ROS family (including ROS); the RET family (including RET); the LTK family (including LTK and ALK); the ROR family (including ROR1 and ROR2); The Musk family (including Musk); the LMR family including LMR1, LMR2 and LMR3); and the SuRTK106 family (including SuRTKΪGό).

Similarly, the serinc /three-nine specific kinases comprise a number of distinct sub- families, including; the extracellular signal regulated kinases, (p42/ERK2 and p44/ERKI); c-Jun NH2-termtnal kinase (JNK); cAMP-responsive element-binding protein kinases (CREBK); c AMP-dependent kinase (CAPK); mitogen-activated protein kinase-activated protein kinase (MAPK and its relatives); stress-activated protein kinase p38/SAPK2; mitogen-and stress-activated kinase (MSK); protein kinases, PKA, PKB and PKC inter alia.

Additionally, the genomes of a number of pathogenic organisms possess genes encoding protein kinases. For example, the malarial parasite Plasmodium falciparum and viruses such as HPV and Hepatitis viruses appear to bear kinase related genes.

Inappropriately high protein kinase activity has been implicated in many diseases resulting from abnormal cellular function. This might arise either directly or indirectly, for example by failure of the proper control mechanisms for the kinase, related for example to mutation, over-expression or inappropriate activation of the enzyme; or by over- or under-production of cytokines or growth factors also participating in the transduction of signals upstream or downstream of the kinase. Tn all of these instances, selective inhibition of the action of the kinase might be expected to ha e a beneficial effect. Diseases where aberrant kinase activity has been implicated include: diabetes; restenosis; atherosclerosis; fϊbrosis of the liver and kidney; ocular diseases; myelo- and lymphoproliferative disorders; cancer such as prostate cancer, colon cancer, breast cancer, head and neck cancer, leukemia and lymphoma; and, auto-immune diseases such as Atopic Dermatitis, Asthma, rheumatoid arthritis, Crohn's disease, psoriasis, Crouzon syndrome, achondroplasia, and thanatophoric dysplasia. The JAK family of protein tyrosine kinases (PTKs) play a central role in the cytokine dependent regulation of the proliferation and end function of several important cell types of the immune system. A direct comparison of the four currently known mammalian JAK family members reveals the presence of seven highly conserved domains (Harpur et al, 1992). In seeking a nomenclature for the highly conserved domains characteristic of this family of PTKs, the classification used was guided by the approach of Pawson and co-workers (Sadowski et al, 1986) in their treatment of the SRC homology (SH) domains. The domains have been enumerated accordingly with most C-terminal homology domain designated JAK Homology domain 1 (JH1). The next domain N-teπninal to Jill is the kinase-related domain, designated here as the JH2 domain. Each domain is then enumerated up to the JH7 located at the N-terminus, The high degree of conservation of these JΛK homology (JH) domains suggests that they are each likely to play an important role in the cellular processes in which these proteins operate. However, the boundaries of the JAK homology domains are arbitrary, and may or may not define functional domains. Nonetheless, their delineation is a useful device to aid the consideration of the overall structural similarity of this class of proteins. Hie feature most characteristic of the JAK family of PTKs is the possession of two ldnase- related domains (JH1 and JH2) ( ilks et al, 1991). The putative PTK domain of JAKl (JH1) contains highly conserved motifs typical of PTK domains, including the presence of a tyrosine residue at position 1022 located 11 residues C-terminal to sub-domain VII that is considered diagnostic of membership of the tyrosine-specific class of protein kinases Alignment of the human JAKl PTK domain (255 amino acids), with other members of the PTK class of proteins revealed homology with other functional PTKs (for example, 28% identity with c-fes (Wilks and Kurban, 1988) and 37% homology to TRK (Kozma et al, 1988)). The JH1 domains of each of the JAK family members possess an interesting idiosyncrasy witt i the highly conserved sub-domain VTIT motif (residues 1015 to 1027 in JAK2) that is believed to lie close to the active site, and define substrate specificity. The phenylalanine and tyrosine residues flanking the conserved tiyptophan in this motif are . unique to the JAK family of PTKs. Aside from this element, the JH1 domains of each of the members of the JAK family are typical PTK domains. Furthermore, there is high sequence identity in the JAK family particularly in and around the ATP binding site (Figure 1). The central role played by the JAK family of protein tyrosine kinases in the cytokine dependent regulati n of the proliferation and end function of several important cell types means that agents which inhibit JAK are useful in the prevention and chemotherapy of disease states dependent on these enzymes. Potent and specific inhibitor-! of each of the currently known four JAK family members will provide a means of inhibiting the action of those cytokines that drive immune pathologies, such as asthma and as immunosuppressivc agents for, amongst others, organ transplants, lupus, multiple, sclerosis, rheumatoid arthritis, psoriasis, Type I diabetes and complications from diabetes, cancer, atopic dermatitis, autoimmune thyroid disorders, ulcerative colitis, Crohn's disease, Alzheimer's disease, and leukcmia/lymphoma. The JAK/STAT Pathway

The delineation of a particularly elegant signal transduction pathway downstream of the non-protein tyrosine kinase cytokine receptors has recently been achieved. In this pathway the key components are: (i) A cytokine receptor chatn.(or chains) such as the Interleukin-4 receptor or the Interferon γ receptor; (ii) a member (or members) of the JAK family of PTKs; (ϋi) a member(s) of the STAT family of transcription factors, and (iv) a sequence specific DNA element to which the activated STAT will bind.

A review of the JAK/STAT literature offers strong support to the notion that this pathway is important for the recruitment and marshalling of the host immune response to environmental insults, such as viral and bacterial infection. This is well exemplified in Table 1 and Table 2. Information accumulated fro gene knock-out experiments have underlined the importance of members of the JAK family to the intracellular signalling triggered by a number of important immune regulatory cytokines. The therapeutic possibilities stemming from inhibiting (or enhancing) the JAK/STAT pathway are thus largely in the sphere of irrtmune modulation, and as such are likely to be promising drugs for the treatment of a range of pathologies in this area. In addition to the diseases listed in Tables 1 and 2, inhibitors of JAKs could be used as immunosuppresive agents for organ transplants and autoimmune diseases such as lupus, multiple sclerosis, rheumatoid arthritis, Type I diabetes, autoimmune thyroid disorders, Alzheimer's disease and other autoimmune diseases. Additionally, treatment of cancers such as prostate cancer by JAK inhibitors is indicated.

Table 1 - Activation of the JAK/STAT pathway in various pathologies

Table 1 (cont.)

Table 2: Diseases Potentially Treatable By JAK-Based Drug Therapies

Table 2 (cont.)

Jak 3 Signalling

Although the other members of the Jak family are expressed by essentially all tissues, JAK3 expression appears to be limited to hematopoetic cells. This is consistent with its essential role in signaling through the receptors for IL-2, 1L4, IL-7, IL-9 and IL-1 by non- covalent association of JAK3 with the gamma chain common to these multichain receptors. Males with X-linked severe combined immunodeficiency (XSC1D) have defects in the common cytokine receptor gamma chain (gamma c) gene that encodes a shared, essential component of the receptors of interleukin-2 (TT.-2), TL-4, TL-7, IL-9, and 1L-15. An XSCID syndrome in which patients with either mutated or severely reduced levels of JAK3 protein has been identified, suggesting that immunosuppression should result from blocking signalling through the JAK3 pathway. Gene Knock out studies in mice have suggested that JAK3 not only plays a critical role in B and T lymphocyte maturation, but that JAK3 is constitutively required to maintain T cell function, Taken together with the biochemical evidence for the involvement of JAK3 in signalling events downstream of the IL-2 and IL-4 receptor, these human and mouse mutation studies suggest that modulation of immune activity through the mhibiton of JAK3 could prove useful in the treatment of T- cell and. B-cell proliferative disorders such as transplant rejection and autoimmune diseases. Prolonged immunomodulation through inhibition of JAK3 signalling should have great therapeutic potential as long as JAK3 inhibition was achieved selectively and not accompanied by inhibition of other kinase-dependent signalling processes. In particular, the high degree of sequence identity held in common by members of the JAK family of kinases raises the possibility that a compound which inhibits Jak3 would also inhibit other members of the family with detrimental long term consequences. For example, prolonged inhibition of Jak2 is likely to lead to erythropenia and (hrorribocytopenia, since the receptors for both crythropoietin and thrombopoietin use only JAK2 for jntraceUular transmission of signals. Selective and Irreversible Inhibition

A PTK catalyses the transfer of a phosphate group from a molecule of ATP to a tyrosine residue located on a protein substrate. The inhibitors known in the art are usually competitive with either the ATP or the protein substrate of the kinase (I-evitzki 2000). Since the concentration of ATP in a cell is normally very high (millimolar , compounds that are competitive with ATP may lack in vivo activity since it is unlikely that said compounds can reach the concentrations within the cell that are necessary to displace the ATP from its binding site.

An alternative approach which has been attempted in relation to EGFR is to design or select compounds which bind to EGFR TK in an irreversible manner. Such compounds are disclosed in Fry 1998; Discafani 1999; SmaiU 1999; SmaiU 2000; Tsou 2001; SmaiU 2001; Wissner 2003. These compounds function as irreversible inhibitors by virtue of the fact that they can form covalent bonds to amino acid residues located at the active site of the enzyme which results in enhanced potency of the compounds in vitro and in the inhibition of growth of human tumors in in vivo models of cancer. A further benefit of such irreversible inhibitors when compared to reversible inhibitors, is that irreversible inhibitors can be used in prolonged suppression of the tyrosine kinase, limited only by the normal rate of receptor turnover. The high homology between members of the JAK family of kinases makes the design of compounds with acceptable selectivity highly challenging. It is believed that by exploiting the minor differences in the amino acid sequence between the members of this family may allow for the identification of selective inhibitors. Alignment of the four members of the JAK family of protein tyrosine kinases reveals that within the amino acids that comprise the ATP-binding pocket of these kinases there are very few amino acid differences that could be used to target potential inhibitors towards one family member or another. Interestingly, JAK3 alone amongst this sub-family of kinases possesses a Cysteine residue close to the front lip of the ATP-binding cavity. It was hypothesised that this may provide a means to develop highly specific irreversible JAK3 inhibitors (Figure 2), by targeting this Cysteine with a functionality bearing an alkylating group such as a Michael acceptor. SUMMARY OF THE INVENTION

The present inventors have found that a group of compounds based upon a disubβtituted heterocyclic scaffold which include an alkylating group such as a Michael acceptor are irreversible and selective inhibitors of the enzyme Janus Kinase 3 and as wiU find appUcations in therapy as i munosuppressive agents for organ transplants, lupus, multiple sclerosis, rheumatoid arthritis, psoriasis, Type I diabetes and complications from diabetes, asthma, atopic dermatitis, autoimmune thyroid disorders, ulcerative colitis, Crohn's disease, and other indications where immunosuppression would be desirable. Furthermore, it is believed that these compounds may find application in therapeutic treatments for proUfcrative diseases and cancers such as Leukemia and Lymphoma where JAK3 is hyperactivated and in diseases such as Alzheimer's disease.

Accordingly, in a first aspect the present invention provides a compound of the general formula I

or pharmaceutically acceptable prodrugs, salts, hydrates, solvates, crystal forms or diastereomers thereof, wherein: Xi, X₂, X₃, X are each carbon where one is substituted with Z a d the rest independently with Y; or one of X,, X^, X₃, , is N, and the others are carbon where one carbon is substituted with Z and the rest independently with Y; A is a ring selected from:

where D is selected from H, C al l, halogen, am o;

Q is a bond, halogen, C_H alkyl, O, S, SO, SO₂, CO, CS;

W is;

(i) NR1R2 where RI and R2 are independently H, C:-, alkyl, Ci-. alkylCF₃, aryl, hetaryl, Ci-j alkylaryl, c_M alkylhetaryl, .₈ cycloalkyl, C₂₆ alkenyl, cyclohetalkyl, Q-i alkylcycloalkyl, C_M alkyl cyclohetalkyl, or RI and R2 are joined to form an optionaUy substituted 3-8 membered ring optionaUy containing an atom selected from O, S, NK3; and R3 is selected from H, -₄ alkyl, aryl, hetaryl, C« alkyl aryl, Cu alkyl hetaryl, COR4 where R4 is selected from H, Q.« alkyl, aryl, hetaryl;

OR

(ii) H, Q-₄ alkyl, aryl, hetaryl, Qι.₈ cycloalkyl, cyclohetalkyl, C alkylaryl, Ci _Λ alkylhetaryl, C₃.₈ cycloalkyl, C₁.₄ alkylcycloalkyl, C_l-4 alkyl cyclohetalkyl;

Y is H, halogen, CN, CF₃, nitro, OH, C_M alkyl, C_H alkylNR5R6, C_l alkylhetaryl, OCn alkyl, OC₂.₄ alkylOC^alkyl, OC_w alkylNR5R6, OQ,₄ alkylhetaryl, OC_1-4 alkykyclohetalkyl, SQ.* alkyl, SC₂-j alkylOC_walkyl, S -₄ alkylNR5R6, NR5R6, NR5COR6, NR5S0₂R6; and E5 and R6 are each independently H, C_M alkyl, or may be joined to form an optionally substituted 3-6 membered ring optionally containing an atom selected from O, S, NR7 and R7 is selected from H, C_M alkyl, aryl, hetaryl, Q- alkylaryl, C__₄ alkylhetaryl;

Z is selected from. :

where RB Is selected from H, C_l-4 alkyl; R9 and RIO are independently selected from H, C_M alkyl, C_MalkylNR12R13, i-i alkylOR12, ι-ι alkylhetaryl or may be joined to form a 5-8 membered ring optionally containing an atom selected from O, S, SO, SO2, NR14; Rll is selected from OH, O _₄ alkyl, NR12R13; n is 0-4; where R12 and R13 are independently selected from H, Gn alkyl, or may be joined to form an optionaUy substituted 3-8 membered ring optionaUy containing an atom selected from O, S, NR14; and R14 is selected from H, C alkyl.

In a second aspect the present invention consists in a composition comprising a carrier and at least one compound of the first aspect of the invention.

I a third aspect the present invention consists in a method of treating a tyrosine kinase- associated disease state, the method comprising administering a therapeutically effective amount oi at least one compound of the first aspect of the invention or a therapeutically effective amount of a composition of the second aspect of the invention.

In a further aspect the present invention provides the use of the compounds of the first aspect or the compositions of the second aspect in the preparation of medicaments for the treatment of JAK3-associated disease states. In a yet further aspect, the present invention provides for a method of suppressing the immune system of a subject, the method comprising administering a therapeutϊca y effective amount of at least one compound, of the first aspect of the invention or a therapeutically effective amount of a composition of the second aspect of the invention, BRIEF DESCRIPTION OF THE FIGURES

Figure 1 shows the amino acid sequence ahgnment of selected Jak Kinases

Figure 2 shows a model of the Jak3 kinase ATP binding pocket displaying the Cysteine residue.

DETAILED DESCRIPTION OF THE [INVENTION Accordingly, in a first aspect the present invention provides a compound of the general formula T

or pharmaceuticaUy acceptable prodrugs, salts, hydrates, solvates, crystal forms or diastereomers thereof, wherein: Xi, X_, X3, X are each carbon where one is substituted with Z and the rest independently with Y; or one of Xi, X₂, X₃, X₄ is N, and the others arc carbon where one carbon is substituted with Z and the rest independently with Y; A is a ring selected from:

where D is selected from H, C_M alkyl, halogen, amino;

Q is a bond, halogen, C_M alkyl, O, S, SO, SO₂, CO, CS;

W s:

(i) NR1R2 where RI and R2 are independently H, C_M alkyl, C_M alkylCF₃, aryl, hetaryl, 0.₄ alkylaryl, C alkylhetaryl, Q_w cycloalkyl, C_2-3 alkenyL cyclohetalkyl, C_w alkylcycloalkyl, C i alkyl cyclohetalkyl, or RI and R2 are joined to form an optionaUy substituted 3-8 membered ring optionally containing an atom selected from O, S, NR3; and R3 is selected from H, C alkyl, aryl, hetaryl, C_M alkyl aryl, CM alkyl hetaryl, C R4 where R4 is selected from H, C alkyl, aryl, hetaryl;

OR

(ii) H, M alkyl, aryl, hetaryl, C_M cycloalkyl, cyclohetalkyl, C alkylaryl, C alkylhetaryl, C₃.B cycloalkyl, C alkylcycloalkyl, C alkyl cyclohetalkyl;

Y is H, halogen, CN, CF₃, nitro, OH, C_M alkyl, C_M alkylNR5R6, C alkylhetaryl, OCM alkyl, OQM alkylOCMalkyl, OCM alkylN 5R6, OCM alkylhetaryl, Od alkylcyclohctalkyl, SCM alkyl, SC₂-₄ alkylOC_Malk l SCM alkylNR5R6, NR5R6, NR5COR6, NR5S0₂R6; and R5 and R6 are each independently H, C alkyl, or may be joined to form an optionally substituted 3-6 membered ring optionally containing an atom selected from O, S, NR7 and R7 is selected from H, C alkyl, aryl, hetaryl, C alkylaryl, C alkylhetaryl;

Z is selected from :

where R8 is selected from H, C alkyl; R9 and RlO are independently selected from H, _M alkyl, Ci .4 alkylNR12R13, C aikyiOR12, C_M aUcylhetaryl or may be joined to form a 5-8 membered ring optionally containing an atom selected from 0, 5, SO, S0₂, NR14; RI 1 is selected from OH, OCM alkyl, NR12R13; n is 0-4; where R12 and R13 are independently selected from H, C alkyl, or may be joined to form an optionaUy substituted 3-8 membered ring optionally containing an atom selected from O, S, NR14; and R14 is selected from H, C alkyl.

In a preferred embodiment the compound is selected from compounds of the general formula II.

π or pharmaceutically acceptable prodrugs, salts, hydrates, solvates, crystal forms or diastcreomers thereof, wherein: Xi, Xz, Xi, X are each carbon where on is substituted with Z and the rest independently with Y; or one of X_L, X₂, X₃, X₄ is N, and the others are carbon where one carbon is substituted with Z and the rest independently with Y; A is a ring selected from:

where D is selected from H, CM alkyl, halogen, amino; Q is a bond, halogen, C alkyl, O, S, SO, SO₂, CO, CS; W is: (i) NR1R2 where RI and R2 are independently H, C alkyl, C_{1 4} alkylCF,, aryl, hetaryl, C alkylaryl, C alkylhetaryl, CM cycloalkyl, C₂._fr alkenyl, cyclohetalkyl, CM alkylcycloalkyl, C alkyl cyclohetalkyl, or RI and R2 arc joined to form an optionaUy substituted 3-8 membered ring optionally containing an atom selected from , S, NR3; and R3 is selected from H, CM alkyl, aryl, hetaryl, Q _A alkyl aryl, C alkyl hetaryl, COR4 where R4 is selected from H, CM alkyl, aryl, hetaryl; OR (ii) W is H, CM alkyl, aryl, hetaryl, Qwj cycloalkyl, cyclohetalkyl. CM alkylaryl, C alkylhetaryl, u> cycloalkyl, C alkylcycloalkyl, C alkyl cyclohetalkyl; Y is H, halogen, CN, CF₃, nitro, OH, C alkyl, C_w alkylNK5R6, CM alkylhetaryl, OCM alkyl, OCM alkylOCMalkyl, OC_M alkylNR5R6, OC alkylhetaryl, OC alkylcyclohetalkyl, SC_M alkyl, SC₂ alkylOC_Malkyl, SCM alkylNR5R6, NR5R6, NR5COR6, NR5S0₂R6; and R5 and R6 are each independently H, C_M alkyl, or may be joined to form an optionaUy substituted 3-6 membered ring optionaUy containing an atom selected from O, S, NR7 and R7 is selected from H, C_M alkyl, aryl, hetaryl, C alkylaryl, CM alkytt etaryl; Z is selected from :

where RS is selected from H, C alkyl; R9 and RIO are independently selected from H, CM alkyl, C alkylNR12R13, C alkylOR12, C alkylhetaryl or may be joined to form a 5-8 membered ring optionaUy containing an atom selected from O, S, SO, SOj, NR14; Rll is selected from OH, OC alkyl, NR12R13; n is 0-4; where: RI 2 and Rl3 are independendy se1 ected from H, C alkyl, or may be joined to form an optionaUy substituted 3-8 membered ring optionally containing an atom selected from O, S, NR14; and R14 is selected from H, C alkyl.

In the above description it will be appreciated that: C alkyl means an unsubstituted or optionaUy substituted straight or branched alkyl chain. Aryl means unsubstituted or optionaUy substituted phenyl or naphthyl. Hetaryl means an unsubstituted or optionaUy substituted 5- or 6-membered heteroaromatic ring containing one or more heteroatoms selected from O, N, S. Cycloalkyl means a 3-8 membered saturated ring, Cyclohetalkyl means a 3-8 membered saturated ring containing 1-3 heteroatoms selected from O, S, NR15, where R15 is H, C alkyl, aryl, hetaryl. Substituents are chosen from halogen, C_w alkyl, CF₃, CN, nitro, aryl, hetaryl, OCF*, OC_M lkyl, OC₂.₅a1kylNRl6Rl7, Oaryl, Ohetaryl, CQ.R16, CONR16R17, nitro, NR16R17, NR16COR17, NR^'16S0₂R17; and R16, R17 are each independently H, C alkyl, C alkyl cycloalkyl, CM alkyl cyclohetalkyl, aryl, hetaryl, Chalky! aryl, C alkyl hetaryl, or may be joined to form an optionally substituted 3-8 membered ring optionally containing an atom selected from O, S, NR18; and R18 is selected from H, CM alkyl, aryl, hetaryl. C alkyl atyl. C alkyl hetaryl.

The compounds of formula I may irreversibly inhibit JAK 3. Generally, the strength of binding of reversible inhibitors of an enzyme is measured by the IC₅₀ value which is a reflection of the equiUbrium constant of the interaction between the inhibitor and the active site of the enzyme. Irreversible inhibitors display an apparent iCm because once the inhibitor is bound it will not leave the active site and the measured ICsu WiU therefore improve (i.e. number will decrease) over time. For instance, the compound f example 20 exhibits an TCα" of ~40nM after 20 minute incubation with enzyme (prior to addition of ATP) whereas the "IC50" drops to 7nM after 90 min pre-incubation.

Preferably, the compound of formul I selectively inhibits JAK 3 with respect to JAK 1 or JAK 2. The term "selectively inhibits" is defined to mean that the apparent IC₅|, of the compound for JAK 3 is more than ten-fold lower (i.e. more potent) than the IC₅₀ for JAK 1 or JAK 2.

The compounds of this invention include all conformational isomers (eg. cis and trans isomers). The compounds of the present invention have asymmetric centers and therefore exist in different enantiomeric and diastereomeric forms. This invention relates to the use of aU optical Isomers and stereoisomers of the compounds of the present invention, and mixtures thereof, and to aU pharmaceutical compositions and methods of treatment that may employ or contain them. The compounds of formula I may also exist as tautυmers. This invention relates to the use of aU such tautomcrs and mixtures thereof.

This invention also encompasses pharmaceutical compositions containing prodrugs of compounds of the formula I. This invention also encompasses methods of treating or preventing disorders that can be treated or prevented by the inhibition of protein kinases, such as JAK comprising administering prodrugs of compounds of the formula I. Compounds of formula I having free amino, amido, hydroxy or carboxyUe groups can be converted into prodrugs. Prodrugs include compounds wherein an amino acid residue, or a polypeptidc chain of two or moire (eg, two, three or four) amino acid residues which are covalently joined through peptide bonds to free amino, hydroxy and carboxyUc acid groups of compounds of formula I. The εurtino acid residues include the 20 naturally occurring amino acids commonly designated by three letter symbols and also include, 4- hydroxyproline, hydroxylysine, demosine, isodemosine, 3-methylhistidine, norvlin,beta- alanϊne, gamma-arninobutyric acid, citrulliiie, homocysteine, homoserine, omifhine and methioine sulfone. Prodrugs also include compounds wherein carbonates, carbamares, amides and alkyl esters which are covalently bonded to the above substituents of formula 1 through the carbonyl carbon prodrug sidechain. Prodrugs also include phosphate derivatives of compounds of formula I (such as acids, salts of acids, or esters) joined through a phosphorus-oxygen bon to a free hydroxyl of compounds of formula L

Where the compound possesses a chiral centre the compound can be used as a purified isomer or as a mixture of any ratio of isomers. It is however preferred that the mixture comprises at least 70%, 80%, 90%, 95%, or 99% of the preferred isomer.

In a stiU further preferred embodiment the compound is selected from the compounds set out in the Examples. More preferably, the compound is selected from the compounds set out in Table 3.

In a second aspect the present invention consists in a composition comprising a carrier and at least one compound of the first aspect of the invention.

In a third aspect the present invention consists in a method of treating a tyrosine kinase- associated disease state, the method comprising administering a therapeuticaUy effective amount of at least one compound of the first aspect of the invention or a therapeutically effective amount of a composition of the second aspect of the invention.

In a further preferred embodiment the disease state involves JAKl, JAK2, JAK3 or TYK2.

In a preferred embcH±Lrnent of the present invention the disease state is selected from the grou consisting of Atopy, such as AUergic Asthma, Atopic Dermatitis (Eczema), and AUergic Rhinitis; Cell Mediated Hypersensitivity, such as Allergic Contact Dermatitis and Hypersensitivity Pneumonitis; Rheumatic Diseases, such as Systemic Lupus Er hematosus (SLE), Rheumatoid Arthritis, Juvenile Arthritis, Sjogren's Syndrome, Scϊeroderma, Polymyositis, Ankylosing Spondyliris, Psoriatic Arthritis; Other autoimmune diseases such as Type I diabetes, autoimmune thyroid disorders, and Alzheimer's disease; Viral Diseases, such as Epstein Barr Virus (EBV), Hepatitis B, Hepatitis C, HIV, HTLV 1, VariceUa-Zoster Virus (VZV), Human PapiUoma Virus (HPV), Cancer, uch as Leukemia, Lymphoma and Prostate Cancer. As used herein the term "tyrosine kinasc-associated disease state" refers to those disorders which result from aberrant tyrosine kinase activity, in particular JAK activity and/or which are alleviated by inhibition of one or more of these enzymes. In a further aspect the present invention provides the use of the compounds described in the preparation of medicaments for the treatment of JAK3-associated disease states.

In a yet further aspect, the present invention provides for a method of suppressing the immune system of a subject, the method comprising administering a therapeuticaUy effective amount of at least one compound of the first aspect of the invention or a therapeuticaUy effective amount of a composition of the second aspect of the invention.

Preferably, the method of suppressing the immune system is for the treatment of disease states selected from lupus, multiple sclerosis, rheumatoid arthritis, psoriasis, Type I diabetes and complications from diabetes, cancer, asthma, atopic dermatitis, autoimmune thyroid disorders, ulcerative colitis, Crohn's disease, and Alzheimer's disease.

Preferably, the method of suppressing the immune system is to modify the immune system response to a transplant into a subject. More preferably, tire transplant is an organ transplant or tissue transplant.

The present invention provides pharmaceutical compositions comprising at least one of the compounds of the formula 1 or II capable of treating a JAK3-associated disorder in an amount effective therefor, and a pharmaceuticaUy acceptable vehicle or dUue t. The compositions of the present invention may contain other therapeutic agents as described below, and may be formulated, for example, by employing conventional solid or liquid vehicles or diluents, as weU as pharmaceutical additives of a type appropriate to the mode of desired adrrtinistration (for example, excipients, binders, preservatives, stabilizers, flavors, etc.) according to techniques such as those well known in the art of pharmaceutical formulation. The compounds of the formula I or il may be administered by any suitable means, for example, oraUy, such as in the form of tablets, capsules, granules or powders; sublinguaUy; buccεύly; parenteraUy, such as by subcutaneous, intravenous, intramuscular, or intracisternal injection or infusion techniques (e.g., as sterile injectable aqueous or non- aqueous solutions or suspensions); nasaUy such as by inlialation spray; topically, such as in the form of a cream or ointment; or rectally such as in the form of suppositories; in dosage unit formulations containing non-toxic, pharmaceutically acceptable vehicles or dUucnts. The compounds may, for example, be administered in a form suitable for immediate release or extended release. Immediate release or extended release may be achieved by the use of suitable pharmaceutical compositions comprising the present compounds, or, particularly in the case of extended release, by the use of devices such as subcutaneous implants or osmotic pumps.

In addition to primates, such as humans, a variety of other mammals can be treated according to the method of the present invention. For instance, mammals including, but not limited to, cows, sheep, goats, horses, dogs, cats, guinea pigs, rats or other bovine, ovine, equine, canine, feline, rodent or murine species can be treated. However, the method can also be practiced in other species, such as avian species (e.g., chickens).

Diseases and conditions associated with inflarnmation and infection can be treated using the method of the present invention. In a preferred embodiment, the disease or condition is one in which the actions of eosinophils and/or lymphocytes are to be inhibited or promoted, in order to modulate the inflarnrnatϋry respoose.

The subjects treated in the above methods, in whom which JAK3 inhibition is desired, are mammals, mduding, but not limited to, cows, sheep, goats, horses, dogs, cats, guinea pigs, rats or other bovine, ovine, equine, canine, feline, rodent or murine species, and preferably a human being, male or female.

The term "therapeuticaUy effective amount" means the amount of the subject composition that will elicit the biological or medical response of a tissue, system, animal or human that is being sought by the researcher, veterinarian, medical doctor or other clinician.

The term "composition" as used herein is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product which results, directly or indirectly, from combination of the specified ingredients in the specified amounts. By "pharmaceutically acceptable" it is meant the carrier, diluent or excipient must be compatible with the other ingredients of the formulation and not deleterious to the recipient thereof. The terms "administration of and or "adirdnistering a" compound should be understood to mean providing a compound of the invention to the individual in need of treatment. The pharmaceutical compositions for the administration of the compounds of this invention may conveniently be presented in dosage unit form and may be prepared by any of the methods weU known in the art of pharmacy. AU methods include the step of bringing the active ingredient into association with the carrier which constitutes one or more accessory ingredients. In general, the pharmaceutical compositions are prepared by uniformly and intimately bringing the active ingredient into association with a Hquid carrier or a finely divided soUd carrier or both, and then, if necessary, shaping the product into the desired formulation. In the pharmaceutical composition the active object compound is included in an amount sufficient to produce the desired effect upon the process or condition of diseases. As used herein, the term "composition" is intended to encompass a product comprising the specified ingredients in the specified amounts, as weU as any product which results, directly or in irectly, from combination of the specified ingredients in the specified amounts.

The pharmaceutical compositions containing the active ingredient may be in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Compositions intended for oral use may be prepared according to any method known to the art for the manufacture of pharmaceutical compositions and such compositions may contain one or more agents selected from the group consisting of sweetening agents, flavoring agents, coloring agents and preserving agents in order to provide pharmaceutically elegant and palatable preparations. Tablets contain the active ingredient in admixture with non-toxic pharmaceuticaUy acceptable excipients which are suitable for the manufacture of tablets. These excipients may be for example, inert dUuents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example, corn starch, or alginic acid; binding agents, for example starch, gelatin or acacia, and lubricating agents, for example magnesium stearate, stearic acid or talc. The tablets may be uncoated or they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glycetyl distearate may be employed. They may also be coated to form osmotic therapeutic tablets for control release. Formulations for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example peanut oil, liquid paraffin, or olive Oil. Aqueous suspensions contain the active materials in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, for example sodium carboxymethy ellulose, methylccUulose, hydroxy- propylmethytceUulose, sodium alginate, polyvinyl-pyrroKdone, gu tragacanth and gum acacia; dispersing or wetting agents may be a naturally-occurring phosphatide, for example ledthin, or condensation products of an alkylene oxide with fatty acids, for example polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyethylene sorbitan monooleate. The aqueous suspensions may also contain one or more preservatives, for example ethyl, or n-propyl, p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose or saccharin.

OUy suspensions may be formulated by suspending the active ingredient in a vegetable oU, for example arachis oil, olive oil, sesame oil or coconut oU, or in a mineral oU such as Hquid paraffin. The ofly suspensions may contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set forth above, and flavoring agents may be added to provide a palatable oral preparation. These compositions may be preserved by the addition of an anti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exempUfied by those already mentioned above. Additional excipients, for example sweetening, flavoring and coloring agents, may also be present.

The pharmaceutical compositions of the invention may also be in the form of oil-in- ater emulsions. The ofly phase may be a vegetable oU, for example otivc oil or arachis oU, or a mineral oU, for example liquid paraffin or mixtures of these. Suitable emulsifying agents may be naruraUy- occurring gums, for example gum acacia or gum tragacanth, naturaUy- occurring phosphatides, for example soybean, ledthin, and esters or partial esters derived from fatty acids and hexitol anhydrides, for example sorbitan monooleate, and condensation products of the said partial esters with ethylene oxide, for example polyoxyethylene sorbitan monooleate. The emulsions may also contain sweetening and flavoring agents. Syrups and elixirs may be formulated with sweetening agents, for example glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain a demulcent, a preservative and flavoring and coloring agents. The pharmaceutical compositions may be in the form of a sterile injectable aqueous or oleagenous suspension. This suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenteraUy-acceptable diluent or solvent, for example as a solution in 1,3-butane diol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic add find use in the preparation of injectablcs.

The compounds of the present invention may also be administered in the f rm of suppositories for rectal adrninistration of the drug. These compositions can be prepared by mixing the drug with a suitable ncm-irritating excipient which is sotid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials are cocoa butter and polyethylene glycols.

For topical use, creams, ointments, jellies, solutions or suspensions, etc., containing the compounds of the present invention are employed. (For purposes of this application, topical appUcation shaU include mouthwashes and gargles.) The compounds of the present invention can also be actirunistercd in the form of liposomes. As is known in the art, liposomes are generally derived from phosphoUpids or other lipid substances. Liposomes are formed by mono- or multUameUar hydrated liquid crystals that are dispersed in an aqueous medium. Any non-toxic, physiologicaUy acceptable and etabolisable lipid capable of forming Hposomes can be used. The present compositions in uposome form can contain, in addition to a compound of the present invention, stabilisers, preservatives, excipients and the like. The preferred lipids are the phosphoUpids and phosphatidyl cholines, both natural and synthetic. Methods to form liposomes are known in the art. The pharmaceutical composition and method of the present invention may further comprise other therapeuticaUy active compounds as noted herein which are usuaUy applied in the treatment of the above mentioned pathological conditions. Selection of the appropriate agents for use in combination therapy may be made by one of ordinary skill in the art, according to conventional pharmaceutical principles. The combination of therapeutic agents may act ynergistically to effect the treatment or prevention of the various disorders described above. Using this approach, one may be able to achieve therapeutic efficacy with lower dosages of each agent, thus reducing the potential for adverse side effects.

Examples of other therapeutic agents include the following: cyclosporins (e.g., cyclosporin A), CTLA4-Ig, antibodies such as ICAM-3, anti-lL-2 receptor (Anti-Tac), anti-CD45RB, anti-CD2, anti-CD3 (OKT-3), anti-CD4, anti-CD80, anti-CD86, agents blocking the interaction between CD40 and gp39, such as antibodies spedfic for CD40 and/or gp39 (i.e., CD154), fusion proteins constructed from CD40 and gp39 (CD4fJlg and CD8gp39), inhibitors, such aa nuclear translocation inhibitors, of NF-kappa B function, such as deoxysperguaUn (DSG), cholesterol biosynthesis inhibitors such as HMG CoA reductase inhibitors (lovastatin and simvastatin), non-steroidal antiinflammatory drugs (NSAIDs) such as ibuprofcn, aspirin, acetaminophen, leflunomide, deoxysperguaUn, azathioprine and cyclooxygenase inhibitors such as r coxib and celecoxib, steroids such as prednisolone or dexamethasone, gold compounds, antiproliferative agents such as methotrexate, FK5Q6 (tacroli us, Prograf), mycophenolate mofetU, cytotoxic drugs such as azathioprine, VP-16, etoposide, fludarabine, dsplatin and cyclophosphamide, TNF-α inhibitors such as tenidap, anti-TNF antibodies or soluble TNF receptor, and rapamycin (sirolimus or Rapamune) or derivatives thereof. When other therapeutic agents are employed in combination with the compounds of the present invention they may be used for example in amounts as noted in the Physician Desk Reference (PDR) or as otherwise determined by one of ordinary skill in the art, Tn the treatment or prevention of conditions which require protein tyrosine kinase inhibition an appropriate dosage level will generally be about 0.01 to 500 mg per kg patient body weight per day which can be administered in single or multiple doses. Preferably, the dosage level wiU be about 0.1 to about 250 g/kg per day; more preferably about 0.5 to about 100 mg/kg per day. A suitable dosage level may be about 0.01 to 250 mg/kg per day, about 0.05 to 100 mg/kg per day, or about 0.1 to 50 mg/kg per day. Within this range the dosage may be 0.05 to 0.5, 0..5 to 5 or 5 to 50 mg/kg per day. For oral adrriinistration, the compositions are preferably provided in the form of tablets containing 1.0 to 1000 UUgrams of the active ingredient, particularly 1.0, 5.0, 10.0, 15.0. 20.0, 25.0, 50.0, 5-0, 100.0, 150.0, 200.0, 250.0, 300.0, 400.0, 500.0, 600.0, 750,0, 800-0, 00.0, and 1000.0 miUigrams of the active ingredient for the symptomatic adjustment of the dosage to the patient to be treated. The compounds ma be administered on a regimen of 1 to 4 times per day, preferably once or twice per day.

It will be understood, however, that the specific dose level and frequency of dosage for any particular patient may be varied and wiU depend upon a variety of factors including the activity of the specific compound employed, the metabohc stabiUty and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the host undergoing therapy.

In order that the nature of the present invention may be more clearly understood, preferred forms thereof wiU now be described with reference to the following non- limiting examples.

EXAMPLES

MATERIALS AND METHODS:

Compound Synthesis Compounds of the general formula I are generally prepared from dihaloheterocycle.

When Q is a bond and W is amino, the synthesis may begin with a nucleophiUc aromatic substitution to generate a monoarr no-monohalo intermediate. The nucleophiUc aromatic substitution is typicaUy carried out by addition of an arr he to the di-halogenated heterαcycle in a solvent such as ethanol, isopropanol, tert-butanol, dioxane, THF, DMF, toluene or xylenc. The reaction is typicaUy performed at elevated temperature in the presence of excess amine or a non-nudeophilic base such as triethylamine or diisopropylethylamine, or an inorganic base such as potassium carbonate or sodium carbonate. Alternatively, the amino substituent may be introduced through a transition metal catalysed amination reaction. Typical catalysts for such transformations include Pd(OAc)₂/P(t-Bu)₃, Fd₂(dba)₃/B1NAP and Pd(OAc)₂/BlNAP. These reactions are typicaUy out in solvents such as toluene or dioxane, in the presence of bases such as caesium carbonate or sodium or potassium tert-butoxide at temperatures ranging from room temperature to reflux. The amines employed in the first step of the synthesis of these compounds are obtained commerctaUy or are prepared using methods well known to those skilled in the art.

When Q is a bond and W is aryl, hetaryl or other simUar carbon-linked systems, the synthesis typicaUy begins with a cross-coupling reaction between dihaloheterocycle and a suitably l^ctionalised coupling partner. Typical coupling partners are boronic adds or esters (Suzuki coupling; see for example Miyaura and Suzuki 1995), sraπnanes (StiUe coupling: sec for example StiUe 1986), Grignard reagents (Kumada coupling: Kumada, Tamao and Sumitani 1988) or orgaπozinc species (Negishi coupling: Negishi 2002). The Suzuki coupling is the preferred coupling method and is typicaUy performed in a solvent such as DME, THF, DMF, ethanol, propanol, toluene, or 1,4-dioxane in the presence of a base such as potassium carbonate, lithium hydroxide, caesium carbonate, sodium hydroxide, potassium fluoride or potassium phosphate. The reaction may be carried out at elevated temperatures and the palladium catalyst employed may be selected from Pd(PPh₃)₄, Pd(OAc)₂, [PdCl₂(dppf)l, Pd₂(dba)₃/P(t-Bu)₃. Where Q is CO, the synthesis begins with the requisite hetaryl caiboxyuc add bearing a halo group. Amide derivatives of the acid may be readily formed by coupling an amine with the acid using coupling reagents such as dicyclc^exylcarbocuxmide, l-(3- dimelhylam oproρyl)-3-ethylcarbodiimide, diisopropylcarbκ>diimidc or carbonyldiimidazole in solvents such as dichloromethane, tetrahydrofuran or 1,4-dioxane. Alternatively, the acid can be converted to the respective acid chloride using thionyl chloride, oxalyl chloride, A^tric-hlorometiTylJcarboriate or eyaπuric chloride, or to the mixed anhydride species using, for example, /-butyl chloroformate, using procedures well known to those skilled in the art. The acid chloride or mixed anhydride derivatives can then be reacted with the desired amine preferably in the presence of abase such as triethylamine, diisopropylethylamine or solid phase equivalent in a solvent such as dichloromethane, tetrahydrofuran, dioxane or ethyl acetate at ambient or elevated temperatures, to generate the amide. The acid chloride may also react with the required at ine under aqueous conditions preferably in the presence of an inorganic base such as sodium hydroxide, potassium hydroxide or sodium carbonate to generate the desired amide. Thioamides may be prepared from the amides formed above by methods weU-known to those skUled in the art and include reaction of the amide with Lawesson's reagent in a solvent such as toluene at elevated temperature. The second step of the synthesis involves a nucleophiUc aromatic substitution reaction of the monohalo intermediate -with a benzimidazole or azabenzimidazole. The reaction is typically performed using a salt of the benzimidazole or azabenzimidazole in solvents such as TΗF, DMF, DMA, NMP, toluene, or xylene from room temperature to reflux. The benzimidazole or azabenzimidazole salt is prepared by reaction with a metal hydride such as sodium or potassium hydride or by reaction With caesium carbonate. Alternatively, a metal-catalysed coupling reaction can be used to introduce the benzimidazole or azabenzimidazole ring- Typical metal catalysts include Pd(OAc)₂/dppf, PdCl₂/dppe, Pd_z(OAc)₂/P(t-Bu)₃, (CuOTt P H. The reaction is typicaUy performed using a base such as caesium carbonate, rubidium carbonate, potassium carbonate, sodium tcrt-butoxide or potassium phosphate in a solvent such as xylene, toluene, or DMF from room temperature to reflux. Auxiliary reagents such as phase transfer agents (e.g. cetrimonium bromide) or copper complexing agents (e.g. phenanthrolinc) may also be employed in the reaction.

Alternatively, the reaction sequence outlined above may be reversed beginning with coupling of the benzimidazole or azabenzimidazole to the dihaloheterocycle using the methods outlined above, foUowed by introduction of the second substituent onto the heterocyclic nucleus using the procedures outlined above.

An alternative route to compounds of the general formula I involves a copper mediated reaction between a benzimidazole or azabenzimidazole and an organometaUic reagent (see for example Finer, 2002). Preferable organometaUic reagents are boronic adds. The thiol reactive moiety (depicted as part of the substituents Z) present in compounds of the general formula I of the invention may be already present in the functionaUti.es employed in the synthetic processes described above or ma be introduced at the final stage of the synthetic procedure. For example, the thiol reactive moiety may be introduced in compounds bearing a free hydroxyl or amino substituent by coupling with a suitable acid. This is typicaUy achieved using coupling reagents such as dicyclohexylcarbodiimide, l-(3-diτnethylammopropyl)-3-efhylcarbodiimidc, diisopropylcarbodiimide or c-ubonyldiimidazole in solvents such as dichloromethane, tetrahydrofuran or 1,4-dioxane, Alternatively, suitable mixed anhydride species of the acid, formed using, for example, butyl chloroformate, using procedures well known to those skiUed in the art, or a suitable acid chloride derivative, can be reacted with the amine or alcohol moiety in the presence of abase such as triethylamine, diisopropylethylamine or soUd phase equivalent in a solvent such as dichloromethane, tetrahydrofuran, dioxane or ethyl acetate at ambient or elevated temperatures, to generate the desired compound-

Those skilled in the art wUl appreciate that the order of the reactions described for the syntheses above may be changed in certain circumstances and that certain functionaUties may need to be derivatised (i.e. protected) in certain instances for the reactions described above to proceed with reasonable yield and efficiency. The types of protecting functionaUry are weU-known to those skiUed in the art and are described for example in Greene (Greene, 1999). The products formed from the reaction sequences described above may be further derivatised using techniques weU known to those skilled in the art.

Representative syntheses are reported below. Example 1 6-CMoro-N-[(TS 'lψΛenylet/tylIpyr 2itt-2-atnme

A solution of ?-α-methy1benzylamine (0.57g, 4.7mmol) and 2,6-dichloropyrazine (0.6388g, 4.29mmol) in dioxane (IB mL) was heated at reflux under Nz for 48 hours. The solvent was removed and the product crystallised from toluene-hexane (0.82g, 82%), ^lH-iuur. (CDG ) δ 1.58 (d,/= 6.6Hz, 3H, CIT₃), 4.88 (m, IH, CH), 5.07 (d, IH, NH), 7.24- 7.36 (m, 5H, Ar-H), 7.61 (s, 1H, pyraz-H), 7.79 (s, IH, pyraz-H). Example 2 N-(tert-øutyV-δ-cfιlσropyra∑irt-2-amwe

A mixture of

(14.9 g, 20 m ot), 2,6-dichloropyraztne (6.0 g, 40 mmol), Hunig's base (lO L) and ethoxyethanol (6 mL) was heated at 13CFC in a sealed tube for 18 hours. The solvent was removed in vacuo and the residue taken up in CH₂Cl₂ (lOOmL) and filtered. The filtrate was washed With H₂0 (2 x 20 mL), brine (20 mL) and dried (Na₂Sθ4). Chromatography eluting with CH₂C1₂ separated the product as a white solid (5.4 g, 72%).

Η-n.m.r. (CDCI3) δ 1.44 (s, 9IT, CH₃), 4.68 (br s, IH, NH), 7.71 (s, IH, pyraz-H), 7.72 (s, IH, pyraz-H).

Example 3

6-Chϊoro-N-f(lS -l-(3-tftefitoxypΑeny ethyUpyra∑in-2-anine

In a procedure analogous to Example 1, reaction of y?-α-me1hylbenzyl_trnirte (l.Og, 6.6mmol) and 2,6-dichtoropyrazine (0.440g, 2.95mmol) furnished the product (517mg, 67%). -n.m.r. (CDC ) δ 1.59 (d, /= 6.9Hz, 3H, CH*), 3.81 (s, 3H, OCIΪj), 4.87 (m, IH, CH), 5.47 (br s, IH, NH), 6.79-730 (m, 4H, Ar-H), 7.66 (s, IH, pyraz-H), 7.79 (9, IH, pyraz-H). Example 4 6-CMoro-N-pJιetιyIpyrazin-2-amine

A solution of 2,6-dichloropyrazine (1 g, 6.7 mmol) and aniline (1.25 g, 13.4 mol) in ethoxyethartol (20 L) containing DIPEA (2.5 mL, 13.4 nunol) was heated at reflux for 3 days under N₃. The solution was concentrated under reduced pressure and the residue diasolvcd in EtOAc (50 L) and washed successively with H₂0 (50 L), 1MHC1 (2 X 50 mL), H₂0 (50 mL) and brine (50 mL). After drying (Na₂S0₄) the solvent was removed under reduced pressure and the residue chromatographed eluting with BtOAc-hexane (20:80 - 50:50) to separate pure product from the lower fractions (230 mg, 17%). Η-n.m.r. (CDCU) 56.62 (br s, IH, NH), 7.11-7.20 (m, IH, ArH), 7.38 (br s, 2H, Aril), 7.40 (s, 2H, ArH), 7.98 (s, IH, pyraz-H), 8.11 (s, IH, pyraz-H).

Example 5

&Chloro-N-f(3^J-7-C4- ethylphenyl)et IJpymzm-2-eim te

In a procedure analogous to Example 1, reaction of α-(. -4-dimethylbenzylarrtine (250mg, 1.85mmol) and 2,6-dichloropyrazine (0.251g, 1.67mmol) furnished the product (199,5mg, 48%).

Η-n.m.r. (CDCk) 81.56 (d, 3H,/= 6.9Hz, CH,), 2.33 (s, 3H, CH₃), 4.84 (m, IH, CH), 5-05 (br s, IH, Nil), 7.15 (AA'XX', 2H, Ar-H), 7.24 (AA'XX', 2H, Ar-H), 7.60 (s, IH, pyraz-H), 7.78 (s, lH, pyraz-H).

Example 6

6-Chloro-N~(4-morpholin-4-ylphenyl)pymzin-2-amine

h a procedure analogous to Example 1, reaction of 4-morpholinoaniline (2.15g, l2.1mmol) and 2,6- ichloropyrazine (0.756g, 5.03mmol) furnished the product (0.54g, 37%).

'H-nmr. (CDCl,) 53.25 (br s, 4H, CH₂), 3.99 (br s, 4H, CH^, 7.05-7.17 (m, 2H, ArH), 7.42- 7.54 (m, 2H, ArH), 7.94 (a, IH, pyraz-H), 8.04 (s, IH, pyraz-H), 8.06 (s, IH, NH). Example 7

6-C?tløro-N-(2f- urylrnettιyl)pymzitι-2-άmitιe

In a procedure analogous to Example 1, reaction of furfurylamine and 2,6- dichloropyrazme furnished the product (98%) .

'H-n.m.r. (CDCl,) 54.57 (d,/= 5.7Hz, 2H, NCHj), 5,01 (s, broad, IH, NH), 6.30 (d,/= 3.3 Hz, IH, furanyl-H), 6.35-633 (m, 2H, furanyl-H), 7.81 (s, III, pyraz.-H), 7.84 (s, IH, pyraz.-H).

Example 8 6-Chlorø-N-(ρyridm-3-ytmet yl)pyr8zin 2-βmme

A mixture 2,6-dichloτopyrazine (0.671 m oi) and 3-picolylamine (2.014 mmol) in xylene (25 ml) was refluxed overnight. The residue obtained after evaporation of the solvent was suspended between CH₂C-₂ (100 ml) and water (100 ml). The organic layer was separated and the aqueous layer was extracted with CH₂C1₂ (3 x 50 ml). The combined organic extracts were washed with brine (1 x 100 ml), dried (Na₂S0₄) and the solvent removed in vacuo. The residue was then purified by column diromatography eluting with a hexane:cthyl acetate gradient mixture to afford the desired product (93%).

Η-n.m.r. (CDCl₃) 84-61 (d,/= 5.7 Hz, 2 H, IVCH_), 5.29 (s, broad, 1H, NH), 7.27 (m, IH, pyrid.-H), 7.30 (m, IH, pyrid.-H), 7.71 (d, /= 7.8 Hz, IH, pyrid.-H), 7.85 (s, IH, pyrid.-H), 8.54= (s, broad, IH, pyraz.-H), 8.61 (s, broad, IH, pyraz.-H). Example 9

N-Benzyl-6-ckiaro-N-^'metlty/pyrazin-2- mme

In a procedure analogous to Example 1, reaction of N-methyl bertzylamine and 2,6- dichloropyrazine furnished the product (70%).

Η-n.m.r. (CDQ,) 83.11 (s, 3 H, NCH₃), 4.78 (s, 2H, ArCH₂N), 7.24 (d,/= 6.9 Hz, 2 H, ArH), 737-728 (m, 4H, ArH), 7.81 (s, IH, pyraz.-H), 7.88 (s, 111, pyraz.-H), Example 10 IH-Beπ∑ifftidazo S-βmme

A solution of 5-nitrobenzimidazole (10.0 g, 61.3 mmol) in methanol (250 mL) was hydrogenated in the presence of 10% Fd/C (0.40 g) at atmospheric pressure for 20h. The mixture was filtered through Celite^® and the solvent removed under reduced pressure to afford the pure product (8.1 g, 100%). Η-n.m.τ. (CD₃OD) 86.75 (dd, lH,/= 8.4 and 2.0 Hz, benzimid-H), 6.92 (d, lH,/= 2.0" Hz, benzimid-H), 7.36 (d, IH,/ = 8.4 Hz, benzimid-H), 7.92 (s, IH, benzimid-H).

Example 11

2-f&(terέ-£ tyiβfrtin&)pynizin-2-y(J-lH-benzimidazoi-δ-a inessά.

7-[6-(tert-faitylffmino)pyrazin-2-yl/-iH-l5'enzimidazol-6 wtiffe

Vrrxx?

A tnixture of lH-benziτrάdazol-5-arxιine (2.93 g, 22 mmol), N-(tert-buryl)-6-chlσropyrazin- 2-amine (3.7lg, 20 mmol) and cesium carbonate (9.12 g, 28 mmol) in DMF (20 mL) was heated under N₂ for 48h. Upon cooling to RT the mixture was filtered and the filtrate concentrated in mcuø. The residue was extracted with CHCI3 and the solvent removed under reduced pressure. The resi ue was chromatographed using CHiClr OH (98;2 - 93:7) to give from the less polar fractions l-[6-(/ butylam o)pyra2m-2-yl]-lH- berιzimtdazol-6-amine (1.38 g):

¹H-n.m.r. (CDQ,) 81.51 (s, 9H,

3-80 (br s, 2H, NH₂), 4.84 (br s, IH, NH), 6.74 (dd, TH,/= 8.4, 2.2Hz, bcnzimid-H), 7.21 (d, lH,/= 2.0Hz, benzimid-H), 7.62 (d, lH,/= 9.2Hz, benzimid-H), 7.79 (s, IH, pyraz-H), 8-07 (s, IH, pyraz-H), 8,17 (s, IH, benzimid-H). and from the more polar fractions H6-( A^-butylamino)pyrazm-2-yl]-lH-benzimidazol-5- amine (1.54 g):

Η-iunx (CDC1₃) δl.51 (s, 9H, C(CH₃)₃), 3.48 (br s, 2H, NH₂), 4.86 (s, IH, NH), 6.79 (dd, lH,/= 8.6, 2.2Hz, benzimid-H), 7.14 (d, lH,/= 2.0Hz, benzimid-H), 7.70 (d, IH, /= 8.6Hz, benzimid-H), 7.78 (9, IH, pyraz-H), 8.07 (s, IH, pyraz-H), 8.47 (s, IH, benzimid-H). Example 12 l-{fc{f(ϊ$)-l-MenyletΛyϊ]ammo/py azin-2-yi)-lH'berιzimidazol-5-amine and l'(6-{[(2S pIιenylethyl/ammσ/pyrazin-2-yl)-lH-be imid zol-6-amifte

To a stirred solution of 5-amino-benzimidazole (290mg, 2.2 mol) in anhydrous DMF (lOmL) under N₂ was added caesium carbonate (980mg) The resulting mixture was stirred at 70°C for 60 min. To this was added a solution of 6-chloro-N-[(lS)-l- phenylcthyl]pyrazin-2-amine (470mg) in DMF (5mL) and the resulting trtixrure was then heated at reflux for 48h. The DMF was removed under reduced pressure and the residue dUuted with chloroform. The organic layer was washed with aqueous Naz O^ dried (Na₂SO*) and the solvent removed under reduced pressure to furnish the crude product. Column chromatography using dichloromethane-methanol (95:5 - 92:8) as eluant separated two fractions from unreactcd starting material. The higher Rf fraction was assigned as the 6-isomer (276mg, 42%). 'H-n.m.r. (CDCl,) 51,64 (d, 3H, /= 6.9Hz, CH₃), 2.90 (br s, 2H, NH₂), 5.05 ( , IH, CH), 5.21 (d, IH, NH), 6.70 (dd, lH,/= 8.7, 2.1Hz, benzimid-H), 6.97 (d, IH, /= 1.8Hz, benzimid-H), 7.28-7.43 (m, 5H, Ph-H), 7.58 (d, IH, /= 8.4Hz, benzimid-H), 7.84 (s, IH, pyraz-H), 8.08 (s, III, pyraz-H), 8.21 (s, IH, benzimid-H). m/z (ES) 331 fJVT+H).

The lower fraction was assigned as the 5-isomer (I70mg, 26%), ¹H-n.m.r. (CDCI3) δl.64 (d, 3H, /= 6.9Hz, CH₃), 2.85 (br s, 2H, NH_), 5.01 (m, IH, CH), 5. 19 (d, IH, NH), 6.70 (dd, IH, /= 8.7, 2.1Hz, benzimid-H), 7.11 (d, lH,/= 1.8Hz, benzimid-H), 7.29-7,40 ( , 5H, Fh-H), 7.51 (d, IH, /= 8.7Hz, benssimid-H), 7.81 (s, IH, pyraz-H), 8.10 (s, IH, pyraz-H), 8.32 (s, IH, benzimid-H). m/z (ES) 331 (M⁺+H). Example 13 l-(6-Chlorσpyrβ∑m~2-yJ)-2H-benzimidazol-5-t}mif3e vϋΛ .l-(6~cfιloropy zin-2-yl)-I'H-bettzimidazol-6-amitte ^{c N N}V

A mixture of lH-bertzimidazol-5-arrune (0.8 g, 6 mmol), 2,6-dichloropyrazine (0.9 g, 6.0 mmol) and cesium carbonate (2.73 g, 8.4 mmol) in DMF (6 mL) was heated under ₂ for 6h. Upon cooling to RT the rruxture was dUuted with dichloromethane-methanol (6:1, 30mL) and filtered and the filtrate concentrated in vacua. The residue was chrυmatographed using F^C -MeOH (98:2 - 94:6) to give from the less polar fractions 1- (6-cWoropyrazm-2-yl)-^"lH-benzimida?'.ol-6-amine (398 mg):

¹H-n.m.r. (CDCI3) & 6.74 (dd, lH,/= 8.2, 2.2Hz, benzimid-H), 7.40 (d, lH,/= 2.2Hz, bcnzimid-H), 7.51 (d, lH,/= 8.2Hz, benzimid-H), 8.40 (s, IH, pyraz-H), 8.48 (s, IH, pyraz- H), 8.83 (s, IH, benzimid-H). and from the more polar fractions l-(6-chloropyrazm-2-yl)-lH-benzimidazol-5-amine (435 mg)

Η-n.m.r. (CDC1₃) 56.79 (dd. 111, /= 8.8, 2.2Hz, l_Jenzimid₇H), 7.03 (d, IH, /- 2.2Hz, benzimid-H), 7.86 (d, IH, /= 9.0 Hz, benzimid-H), 8.44 (s, IH, pyraz-H), 8.52 (s, IH, • pyraϋ-H), 8.82 (s, IH, benzimid-H). Example 14

1-f&[(Cy opropyimethyl)anιinσIpyrazm-2-yl/--lJI-l)efizi idazoI-&a e

A solution of l-(6-chloropyrazin-2-yl)-lH-benzimidazol-6-arnine (100 mg, 0.41 mmol) and cyclopropylmethylamine (424 μL, 4.1 mmol) in ethoxyethanol (2 L) containing DTPEA (140 μL) was heated at reflux overnight under N₂. The solution was concentrated under reduced pressure and the residue dissolved in EtOAc (20 mL) and washed successively ith H₂0 (20 mL), IM HCl (2 x 20 L), H₂0 (20 L) and brine (20 mL). After drying (Na₂S0₄) the solvent was removed under reduced pressure and the residue chromatographed eluting with dichloromethane-methanol (9:1 - 94:6) to separate pure product from the lower fractions (98 mg)

¹H-n.m.r. (CDCL) δ .28-0.36 (m, 2H, CH₂), 0.57-0.66 (m, 2H, CH₂), 1.08-1.22 (m, IH, CH), 3.27-3.34 (m, 2H, CH₂), 3.79 (br s, 2H, NH₂), 5.02 (m, IH, NH), 6.74 (dd, lH,/= 8.6, 2.2Hz, benzimid-H), 7.33 (d, IH; /= 2,2Hz, benzimid-H), 7.61 (d, IH, J = 9.2Hz, benzimid-H), 7.84 (s, IH, pyraz-H), 8.10 (s, IH, pyraz-I I), 835 (s, IH, benzimid-H),

Example 15 l-[6-dsopropylamino)pyrazin-2-yl/-lH-benzi idazol'6-amifie

A solution of l-(6-cWoropyrazin-2-yl)-lH-benzirmdazol-6-amine (100 mg, 0.41 mmol) and isopropylamine (350 μL, 4,1 mmol) in ethoxyethanol (2 mL) containing DIPEA (140 μL) was heated in a sealed tube overnight under N₂. The solution was concentrated under reduced pressure and the residue d issolved in EtOAc (20 mL) and washed successively with H₂0 (20 L) and brine (20 mL). After drying ( ₂Sθ₄) the solvent was removed ^* I under reduced pressure and the residue chromatographed eluting with dichloromethane- methanol (9:1 - 94:6) to separate pure product from the lower fractions (102 mg).

Η-n.m.r. (CDCI3) δl.33 (d, 6H, /= 6.4Hz, CH₃), 3.79 (br s, 2H, NH₂), 4.05-4.21 (m, III, CH), 4.72 (m, IH, /= 7.2Hz, NH), 6.75 (dd, lH,/= 8.6, 2.2Hz, benzimid-H), 7.32 (d, lH,/= 2.0Hz, benzimid-H), 7.61 (d, lH,/=* 8.4Hz, benzimid-H), 7.79 (s, IH, pyraz-H), 8.09 (s, IH, pyraz-H), 8.35 (s, IH, benzimid-H).

Example 16 7-[6-(Diethyl inopyrazin-2-ylJ-lH-beni!imidazol-6-ιmιme

A solution of l-(6-chloropyrazm-2-yl)-lH-benzimidazol-6-anτine (100 mg, 0.41 mmol) and dicthylamine (430 μL, 4.1 mmol) in ethoxyethanol (2 mL) containing DIPEA (1 0 μL) was heated in a sealed tube overnight under N₂. The solution was concentrated under reduced pressure and the residue dissolved in EtOAc (20 mL) and washed successively with H_zO (20 L) and brine (20 L). After drying (Na₂SO,,) the solvent was removed under reduced pressure and the residue chromatographed eluting with dichloromethane- methanol (9:1 - 94:6) to separate pure product from the lower fractions (110 mg).

'H-n.m.r. (CDCl.) 51.28 (t, 6H,/= 7.1Hz, CH3), 3.61 (q, 4H, /= 7.1Hz, Cti₂), 3.78 (br s, 2H, NH₂), 6.74 (dd, lH,/= 8.6, 2.2Hz, benzimid-H), 732 (d, HT,/= 2.4Hz, benzimid-H), 7.61 (d, HI, /= 8.8Hz, bervzimid-H), 7.91 (s, IH, pyraz-H), 8.06 (s, III, pyraz-H), 8.36 (s, IH, berizimid-H). Example 17 l-(&-Pyridin-4-ylpyraz '2-yl)-lH-nenzimidazoi-6-amine

Under a nitrogen atmosphere a mixture of l-(6 hloropyrazin-2-yl)-lH-bcnzimidazol-6- amine (50 mg, 0.20 mmol), 4-pyridylboronic add (30 mg, 0.24 mmol), tβtralds(triphenylphosphine)paUadium(0) (23 mg, 0.02 mmol) in toluene — -propanol (2 mL, 3:1) was treated with 2M aqueous sodium carbonate solution (0.14 mL, 0.84 mmol). The resulting mixture was stirred vigorously whilst being heated under reflux overnight. Upon cooling, the mixture was diluted with ethyl acetate (10 mL) and washed with H₂0 (1 x 10 mL). The aqueous phase was extracted with ethyl acetate (10 mL) and the organic layers combined and washed with 0.5M NazCOs, brine and then dried (Na₂S0₄). Removal of solvent i vac o then yielded crude product, which was purified by column chromatography using dichloromethane-methanol (98:2 - 91:9) as cluent to furnish the product (32 mg).

'H-n.m.r. (CDCl,) δ 3.88 (s, broad, 2H, NH²), 6.80 (dd, lH,/= 8.6 and 2.0 Hz, benzimid-H), 7.46 (d, IH,/- 2. Hz, benzimid-H), 7.67 (d, 1H, /= 8.6 Hz, benzimid-H), 7.98 - 8.01 (m, 2H, pyrid-H), 8,49 (s, IH, pyraz-H), 8.84 - 8.87 (m, 2H, pyrid-H), 8.99 (s, IH, pyraz-H), 9.05 (s, IH, benzimid-H).

Example 18 N-f7-I&-(eεrt-β t lamino)7ψrazitt-2-ylJ-lH-benzimidazol-6-yl}prop-2-ynamide

To a stirred solution of l-l6-(tert-butylarnmo)pyraz -2-ylJ-lH-benziιrύdazol-6-amine (70 mg, 0.25 mmol) in anhydrous dichloromethane (2.5 mL) under N₂ was added triethylamine (86 μl), EDAC.HC1 (60 mg), 4 (l-pyrroHdinθ)pyridine (4 mg) and propioUc acid (18.5 μL). The resulting mixture was then stirred at RT overnight and was the dUuted with CH₂C1₂ (lO L) and washed with H_aO (2 x 10 mL), 0.5M Na₂C0₃ (10 mL) and dried (Na₂S0₄). The solvent was removed under reduced pressure and the residue was purified by column chromatography using dichloromethane-methanol (99:1 - 91:9) as eluant to separate the pure product (1.8 mg). ¹H-n-m.r. (CDCL) 51.52 (s, 9H, CH₃), 4.76 (br s, IH, NH), 5.78 (br s, H, CH), 6.75 (dd, IH, /=8.4, 2.2 Hz, ArH), 7.22 (d, lH,/= 2.2Hz, ArH), 7.63 (d, 1H,7= 8.0Hz, Λr-H), 7.79 (s, IH, pyraz-H), 8.08 (s, IH, pyraz-H), 8.37 (s, IH, benzimid-H).

Example 19

N-fl-(6^i(R)-l-Pkenyiethylkminofpyraziιt-2-yi jFf-benzimidazσI-6-y- Uacryiamide

To a stirred solution of l-(6-{[(l-^-l-phenylemylJaιrιmo}pyrazin-2-yl)-"lH-ben_--imidazol-6- arnine (67 mg, 0.2 mmol) in anhydrous THE (2mL) under N₂ was added triemylatriine (67 μL 0.48 mmol), EDAC JICl (46 mg, 0.24 mmol), 4-(l-pyrrolidino)pyridine (cat.) and acrylic acid (17 g, 0.24 mmol). The resulting mixture then stirred at RT overnight and was the dUuted ith H₂0 (10 mL) and the mixture extracted with EtOAc (2x 10 mL). The combined organic layers were washed with saturated aqueous aaCOj, dried (Na₂S ₄) and the solvent removed in vacuo. The residue was purified by column chromatography using dichloromethane-metlianol (98:2 - 94:6) as eluant to separate the pure product (25 g). 'H-n-rr r. (CDCla) §1.62 (d, 3H, J = 6.8Hϋ, CH₃), 5.01-5.13 (m, IH, CH), 5.38 (d, IH, /= 6.4Hz, NH), 5.78 (dd, lH,/= 9.8, 2.0Hz, CH), 6.24-6.52 (m, 2H, 2 x CH), 7.29-7.44 (m, 6H, ArH), 7.70-7.74 (m, 2H, Ar-H), 7.82 (s, IH, pyraz-H), 8.11 (s, IH, pyraz-H), 8.33 (s, IH, benzimid-H), 8.42 (s, IH, CONH). Example 20 N-/7-[6-(tert-Biityl nino)pymzin-2-yif-ΪH-benzimidazoi-6-yi/acryiamide

To a stirred solution of l-[6-(tcrt-burylammo)pyrazm-2-yl]-lH-rjenzimidazol-6-amrne (22 mg, 0.08 mmol) in anhydrous dichloromethane (2mL) under N₂ was added tricmylamine (33 μL, 0.24 mmol), EDACHC1 (22 mg, 0.12 mmol), 4-(l-pyrrolid o)pyridine (cat.) and acrylic acid (8 μL, 0.12 mmol). The resulting mixture then stirred at RT for 3 days and was the diluted with H₂ (10 mL), the organic phase separated and the aqueous phase extracted with CH₂C1₂ (10 mL). The combined organic layers were dried (Na₂S0₄) and the solvent removed in vacuo. The residue was purified by column chromatography using dldiloromethane-methanol (98-2 - 93:7) as ehiant to separate the pure product (10 mg).

'H-n.m.r. (CDCL) 31.50 (s, 9H, CH₃), 4.89 (br s, IH, NH), 5.77 (dd, IH, = 10.0, 2.0Hz, CH), 6.24-6.51 ( , 2H, 2 x CH), 7.25 (dd, 1 H, /-= 8.6, 2.0Hz, ArH), 7.76 (d, IH, = 8.8Hz, Ar-H), 7.83 (s, IH, pyraz-H), 7.88 (br s, IH, CONH), 8.13 (s, IH, pyraz-H), 8.52 (s, IH, benzimid- H), 8.56 (s, IH, ArH).

Example 21 N-{l-f6-(tert-Butyhntino)pyrβz -2-yiJ-lH-benzimidazόl-5-y(/acryia ide

To a stirred solution of l-[6-(tert-butylammo)ρyrεrzm-2-yl]-lH-benzimidazol-5-amine (20 mg, 0.08 mmol) in anhydrous dichloromethane (2 mL) under N₂ was added triethylamine (33 μL, 0.24 mmol), EDAC.HC1 (22 mg, 0.12 mmol), 4-(I-pyιϊ lidmo)pyridine (cat.) and acryUc acid (8 μL, 0.12 mmol). The resulting mixture then stirred at RT for 3 days and was the diluted with H₂0 (10 mL), the organic phase separated and the aqueous phase extracted with CH₂C1₂ (10 mL). The combined organic layers were dried (Na₂S0₄) and the solvent removed in vac o. The residue was purified by column chromatography using dichloromethane-methanol (982 - 92:8) as eluant to separate the pure product (10 mg). -n.m.t. (CDCL.) 51-52 (s, 9H, CH₃), 4-87 (br s, IH, NH), 5.77 (dd, lH,/= 9.8, 2.0Hz, CH), 6.31 (dd, 1H,7= 16.6, 9.8Hz, =CH(H)), 6.48 (dd, IH,7= 16.6, 2.0Hz, =CH(H)), 7.73-7.81 (m, 2H, pyraz-H + ArH), 7.89 (d, lH,./= 8.8Hz, ArH), 8.01 (s, IH, ArH), 8.10 (s, IH, pyraz-H), 8.55 (s, 1 H, benzimid-H). Example 22

N-/l-l6-fterf-Bufyh ino)py zitt-2-yl/-2H-l?enzimidazo/-S lJ-2-ntethy!acη ia ide

Following a procedure identical to Example 21 however using methacrylic a id in place of acrylic acid, l-[6-(tert-butylarmno)pyraz -2-ylJ-lH-benzirιudazol-5-arrtine (57 mg) afforded N-{l-[6-(tert-butylaιnmo)pyrazin-2-yl]-lH-benzimidazol-6-yl}-2- methylacrylamidc (54mg).

Η-ιun.r. (CDC.₃+ di-MeOD) 51.43 (s, 9H, CH₃), 2.00 (br s, 3H, CH₃), 5.42 (br s, IH, =CH(H)), 5.77 (br a, IH, =CH(H)), 7.32 (dd, IH, /= 8.2, 2.0 Hz, ArH), 7.67 (d, 1H,7= 8.8 Hz, ArH), 7.74 (s, ]H, pyraz-H), 7.99 (s, IH, pyraz-H), 8.38 (d, 1H,/= ZO Hz, ArH), 8.46 (s, IH, beri-zimid-H).

Example 23 J~(6-i(2-Methyiphenyiarnin Jpyrazifi-2-ylJ-I^' -l>enzimida∑ol-6-iintine

To a stirred solution of the chloropyrazine (100 mg, 0.40 mmol) in toluene (2 mL) was added ø-toluidinc (0.1 mL, 0.93 rrttnol), Pd[P(/-Bu)₃]₂ (10 mg) and sodium /-butoxide (58 mg, 0.6 mmol). The solution was heated at 80° overnight and upon coohng to RT was dUuted with EtOAc (20 L). The organic layer was collected and the aqueous layer extracted with EtOAc (20 mL) and the combined organic layers washed with water, brine, and dried ( a₂Sθ ). Removal of the solvent under reduced pressure gave an oily residue which was chromatographed using CH2C12-MeOH (98:2 -> 94:6) to separate the desired product as a pale yellow oil (52 mg, 41%). ^xH-n.m.r. (CDC1₃) 52.34 (s, 3H, CH₃), 3.70 (s, 2H, NH₂), 6.61 (s, IH, NH), 6.71 (dd, IH, /= 8.6, 2.2 Hz, ArH), 7.19-7.36 (m, 4H, ArH), 7.53-7.60 (m, 2H, ArH), 7.99 (s, IH, pyraz-H), 8.27 (s, IH, pyraz-H), 8,36 (s, IH, benzimid-H).

Example 24

(22J'N-fl^/6-&&τX~Butyiammσ)ρymzin-2-yif-2If-benzimidazof-6-yiJ-3 ^^ ylacrylamide

Y

To a stirred solution of the alkyne (30 mg, 0.07 mmol) in anhydrous ethanol (5 mL) was added Lindlar catalyst (3 mg). The mixture was then purged with hydrogen gas and stirred under H₂ at atmospheric pressure or 3h. The catalyst was removed by filtration through Celite^® and the solvent removed in vac'M. Flash chromatography using EtOAc- MeOH (9:1) separated pure product as a sticky semi-soUd (13 mg, 43%). ^xH-n.m.r. (CDCL) 51-50 (s, 9H, C(CH₃)₃), 4.93 (s, IH, NH), 6.26 (d, IH, /= 12.6Hz, C=CH), 6.82 (d, IH,/_* 12.6Hz, C=CH), 7.14(dd, lH,/= 8.7, 2.1Hz, ArH), 7.25-7.29 (m, IH, pyr idine-H), 7.74 (d, IH, /= 8.7Hz, ArH), 7.82 (s, IH, pyraz-H), 8.07 (br s, IH, CONH), 8.09 (s, IH, pyraz-H), 8,13-8.16 (m, IH, pyridine-H), 8.47 (d, !H,/= 1.8Hz, pyridine-H), 8.50 (s, IH, benzimid-H), 8.51-8.53 (m, IH, pyridine-H), 8.63 (d, IH, J = 2.1Hz, ArH). m/z (El): 413 (-vT). Example 25 M-(tet\.--Buty0-6-<chioropyrazirte-2-carbi>x(i?nide

Thionyl chloride (1 L, 13.7 mmol) was added to a suspension of the acid (315 mg, 2 mmol) in toluene (5 mL). A drop of DMF was then added and after stirring at RT for 10 min. the mixture was heated at reflux for Ih. The reaction was cooled to RT and toluene and excess thionyl chloride were removed under reduced pressure. Toluene (1 L) was then added to the residue and this was removed under reduced pressure. This process was repeated, and then CT^CL. (10 L) was added and the resulting solution cooled to 0°C. t-Butylamine ( 0.45 mL, 4.3 mmol) and triethylamine (l.lmL, 8.0 mmol) were then added and the solution stirred at RT overnight. The solution was diluted with CH₂Cl (10 mL) and H₂0 (10 mL) and the organic layer coUected and washed with aq. Na₂C0₃ and then dried (Na₂SO<). The solvent was removed in vacua and flash chromatography of the residue using CH₂CI₂-MeOH (95:5) separated the pure product as an oU (290 mg, ,68 %). H-n.m.r. (CDCU) 61.49 (s, 9H, C(CH₃)₃), 7,48 (br s, IH, NH), 8.72 (s, IH, pyra .-H), 9.27 (s, IH, pyraz-H). m/z (El): 413 (IvT). Example 26 l-(6-Methσxypyridin~3-y/)-5-nilτa~lH-hettzmιidazoie∞\ά J-(6-^'tnetiιoxypyridm-3-yl)-6- nitro-l -ben∑imidσzole

A mixture of 5-nitrobenzirrtidazoIe (650 mg, 4 mmol), 2-methoxy-5-pyridyl boronic add (420 mg, 2.6 mmol), copper (II) acetate (1.09 g, 6 mmol) and powdered 4A sieves was stirred vigorously in CH₂C1₂ (40 L) containing pyridine (0.65 mL) was stirred in the air over 3 days. The mixture was then filtered through CeHte^® and the filter pad washed with CH_{2 2}-Me H (4:1). The filtrate and washings were combined, concentrated // vacua and the residue chromatographed using CHaCl₂-MeOH (100:0 -> 95:5) to separate the product (as a 1:1 mixture of regiomers) as a white solid (272 mg, 37 %). m/zφX): 270 (M ). Example 27 l-(6-Metkoxyρyridin'3-y -2N-benzι idazoi-5-amine&xv 2-(&met7ιoxypyridin-3-yl)-J - benzimidazol-6'amine

The mixture of regioisomers derived from Example 26 (270 mg, 1 mmol) was hydrogenated following the procedure outlined in Example 1 . The crude product was chTOmatographed eluting with CH₂ClrMe H (98:2 ^■> 95:5) to separate the 6-isomer (84 mg)^*from the less polar fractions and the 5-isomcr from the polar fractions. (122 mg).

6-isomer: H-n.m.r. (CDCh) δ 3.88 (br s, 2H, NH₂), 4.01 (s, 3H, OCH,), 6.64 (d, lH,/= 2.1Hz, benzimid-H), 6.72 (dd, lH,/=* 8.7, 2.1Hz, benzimid-H), 6.92 (d, IH,/- 9.0Hz, benzimid- H), 7.61-7.68 (m, 211, pyr-H), 7.82 (s, IH, beruΛtmid-H), 8.30 (d, lH,/= 2,7Hz, ρyr-H).

5-isomer: -n.m.r. (CDCI3) 63.11 (br 5, 2H, NH₂), 4.01 (s, 3H, OCH₃), 6.75 (dd, lH,/= 8.4, 2.1Hz, bcnzimid-H), 6.92 (d, IH, /- 8.7Hz, benzimid-H), 7.15 (d, IH,/-_* 2.1Hz, benzimid-H), 7.18 (d, lH,/= 8.7Hz, pyr-H), 7.68 (dd, lH,/= 8.7, 2.7Hz, pyr-H), 7.91 (s, IH, benzimid-H), 8.31 (d, lH,/= 2,7Hz, pyr-H). Example 28 l-(5-βromopyridin-3-yl)-JFi-benzmtidazol-6-arnittesύaά. 7-(5-brornopyridin-3-yi)-7H~ benzmtidazol-5-a?nine

A solution of 3,5-dibromopyridine (2.37g, 10 mmol), 5-am obenzimidazole (l.όOg, 12 mmol) and caesium carbonate (4.9g, 15 mmol) in DMSO (10 mL) was heated at 150" for 18h. Upon cooling to RT the solution was diluted with CHCL, (40 mL) and filtered through Celite^® and the filtrate concentrated in vacua. The residue was chromatographed (pre-adsorption to s ica) eluting with EtOAc-MeOH (100:0 -> 95:5) to separate, from the less polar fractions, ^ne 6-isomer, and from the more polar fractions the 5-isomer.

6-isomer: -n.m.r. (CDCL) δ 3.82 (br s, 2H, NH*), 6.75-6.78 (m, 2H), 7.64 (d, lH,/= 9.0Hz, benzimid-H), 7.89 (s, IH), 8.01 (dd, lH,/= 2.1Hz, pyr-H), 8.75 (br s, 2H).

5-isomer: ¹H-n.m.r. (CDCL) 53.74 (br s, 2H, NH₂), 6.79 (dd, IH, /= 8.7, 2.1Hz, ben/tmid-H), 7.15 (d, IK,/- 2.1Hz, benzimid-H), 7.31 (d, IH,/« 8.7Hz, benzimid-H), 7.99 (s, IH, benzimid-H), 8.01 (dd, lH,/= 2.1, 2.1Hz, pyr-H), 8.74-8.77 (m, 2H, pyr-H). Example 29 2-(&Brø?nopyridin-2-y7)-lH-aenzimidazoi-6-atniπe and l~(6-bronwpyrirfm-2-ytYlH- benzimidazai-5-amine

Using identical procedures to those outlined in Example 28, reaction of 2,6- dibromopyridme with 5-am σbenzimidazole and caesium carbonate in DMSO at 150" afforded the two regioisomeric products whic were separated by chromatography.

6-isomer:

Η-n-m.r. (CDCL) δ 3.83 (br s, 2H, NH₂), 6.75 (dd, lH/= 8.4, 2.1Hz, benzimid.-H), 7.42- 7.47 (m, 3H), 7.60 (d, lH,/= 8.4Hz), 7.71 (dd, IH, /= 7.8Hz), 833 (s, IH).

5-isomer:

Η-n.m.r. (CDCL,) 56.81 (dd, lH,/= 8.7, 2.1Hz, benzimid-H), 7.12 (d, lH,/= 8.1Hz, bertzimid-H), 7.40-7.48 (m, 2H), 7.70 (dd, lH,/= 7.8, 7.8Hz, pyr-H), 7.89 (d, lH,/= 8.7Hz, benzimid-H), 8.46 (s, IH).

The following compounds were prepared using analogous procedures to those described above:

6.36-

(s,

(dd,

/= IH, (s,

311, 6.92

IH, (s, (m,

(d,

(»,

9H, 2 X

IH,

8.46

6H,/ 9H, 2 x 1H, (d, (s, /= (s,

IH),

21i), (b,

9H, IH, (dd, 7.83 IH, lH,

SCREENING Compound Dilution For screening purposes, compounds Were diluted in 96 well plates at a concentration of 20 μM. Plates were warmed at 37^PC for 30 minutes before assay. JAK Tyrosine Kinase Domain Production AK kinase domains were produced in the following manner^*. /A 1 T ie kinase domain of human JAKl wag amplified from U937mRN A using the polymerase chain reaction with the following primers: XHOI-jl 5'-CCG CTC GAG ACT GAA GTG GAC CCC ACA CAT-3' J1-KPN7 5'-CGC GGT ACC TTA TTT TAA ΛAG TCC TTC AAA-3' JAKl PCR products were cloned into the pFastBac HTb expression vector (Gibco) via the Xho 1 and Kpn I sites. The JAKl plasmid was then transformed into competent DHlOBac cells (Gibco), and the recombinant baculovirus produced prepared for transfection into Sf9 insect cells. JA 2

The kinase domain of humanJAK2 was amplified from U937mRNA using the polymerase chain reaction with the following primers:

SALI-jk2 5'-ACG CGT CGA CGG TGC CTT TGA AGA CCG GG A T-3' jk2-NOTT 5'-ATA GTT TAG CGG CCG CTC AGA ATG A AG GTC AIT T-3' JAK2 PCR products were cloned into the pFastBac HTc expression vector (Gibco) via the Sal I and Not 1 sites. The JAK2 plasmid was then transformed into competent DHlOBac cells (Gibco), and the recombinant baculovirus produced prepared for transfection into Sf9 insect cells. JAK3 The kinase domain of humanJAK3 was amplified from U937mRNA using the polymerase chain reaction with the following primers: XHOI-J3 5'-CCG CTC GAG TAT GCC TGC CAA GAC CCC ACG-3' J3-KPNI 5'~CGG GGT ACC CTA TGA AA A GGA CAC GGA GTG-3' JAK3 PCR products were cloned into the pFastBac HTb expression vector (Gibco) via the Xho I and Kpn 1 sites. The JAK3 plasmid was then transformed into competent DHlOBac cells (Gibco), and the recombinant baculovirus produced prepared for transfectiαn into Sf9 insect cells. TY 2 The kinase domain of humanTYK2 was amplified from A549 mRNA using the polymerase chain reaction with the following primers: HT2FK 5'-GGA GCA CTC GAG ATG GTA GCA CAC AΛC CAG GTG-3' ITΥ2.2R 5'-GGA GCA GGA ATT CCC GCG CTG CCG GTC ΛA A TCT GG-3' TYK2 PCR products were cloned into ρBlueBacHis2A (Invitrogen) via the EcoRT site. The recombinant TYK2 baculovirus produced was prepared for transfected into Sf9 insect cells.

Large Scale Production Of Kinase Domains Baculovirus preparations from each of the JAK family members were infected into five litres of High Five cells (Jhvitrogen) grown in High Five serum free medium (Invitrogen) to a cell density of approximately 1-2 X 10⁶ cells/ml. Cells are infected with virus at a MOI of 0.8-3.0. Cells ere harvested and lysed. JAK kinase domains were purified by affinity chromatography on a Probond (Invitrogen) nickel chelate affinity column. Assay Protocols

Kinase assays were performed either in a 96 well capture-based ELISA assay or in 384 well Optiplates (Packard) using an Alphascreen Protein Tyrosine Kinase kit. In either casse using approximately 1.5 μg of affinity purified PTK domain in the presence of 50mM HEPES, pH 7.5, lOrnM MgClj, lSOrnM NaCl and lOμM-lmM ATP. The biotinylated substrate biol n-EGPWLEEEEEAYGWMDF-NHi (final concentration 5μM) was used as substrate. In the ELISA assay tyrosine phosphorylation was quantitated following transfer to an avidin coated ELISA plate using peroxidase-linked anti-phospho-tyrosine antibody FY20. hi the Alphascreen assay, Alphascreen phosphσtyrσsinc acceptor beads followed by streptavidin donor beads were added under subdued light. The HLISA plates were read on a BMG Fluσrostar, the Alphascreen plates were read on a Packard Fusion Alpha. Inhibitors were added to the assays fifteen minutes prior to the addition of ATP. Inhibitors were added in aqueous DMSO, with DMSO concentrations never exceeding 1%. Results The activity of selected compounds is shown in Table 3. Compounds that exhibited a capacity to inhibit 50% of JAK activity at a concentration of 20μM (measured under standard conditions, see Methods), are designated as "+". Table 3

Table 3 (cont.)

Throughout this specification the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps. All publications mentioned in this specification are herein incorporated by reference. Any discussion of documents, acts, materials, devices, articles or the like which has been included in the present specification is solely for the purpose of providing a context for the present invention. It is not to be taken as an admission that any or all of these matters form part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed in Australia or elsewhere before the priority date of each claim of this application.

It will be appreciated by persons skilled in the art that numerous variations and/or modifications may be made to the invention as shown in the specific embodiments ithout departing from the spirit or scope of the invention as broadly described. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive.

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2. Finet, J.-P., Fedorov, A.Y., Combes, S., and Boyer, G. (2002) Recent Advances in Ullmann Reaction : Copper (II) Diacetate Catalysed At-, O- and £■ Arylation Involving Polycoordinate Heteroatomic Derivatives. Curr. Org. Chem. 6, 597-626. 3. Fry DW, Bridges AJ, Denny WA, Dohcrty A, Grcis KD, Hicks JL, Hook KE, Keller PR, Leopold WR, Loo JA, McNamara DJ, Nelson JM, Sherwood V, Smaill JB, Trumpp- Kalltneyer S, and Dαbrusin EM. (1998) Specific, irreversible inactivation of the epidermal growth factor receptor and erbB2, by a new class of tyrosine kinase inhibitor. Proc NatlΛ d Sci USA. 95, 2022-7. 4. Hovens CM, Stacker SA, Andres AC, Harpur AG, 2_icmiecki A, and Wilks AF. (1992) RYK, a receptor tyrosine kinase-related molecule with unusual kinase domain mo ifs. Proc NatiAcad Sci USA. 89, 11818-22.

5. Koz a SC, Redmond SM, Fu XC, Saurer SM, Groner B, and Hynes NE. (1988) Activation of the receptor kinase domain of the trk oncogene by recombination with two different cellular sequences. EMBOf, 7, 147-54.

6. Kumada, M.; Tamao, K.; Sumitani, K. (1988) Phosphine-Nickel complex catalysed cross-coupling of Grignard reagents with aryl and alkenyl halides: 1,2- Dibutylbenzene. Org. Synth. Coil. Vol.6, 07. 7. Levitz i A. (2000) Protein Tyrosine Kinase Inhibitors as Therapeutic Agents. Top. Curr. Chem.211, 1 -15.

8. Miyaura, N. and Suzuki, A. Palladium-Catalyzed Cross-Coupling Reactions of Organoboron Compounds (1995) Chem Rev.95, 2457 9. Negishi, E. (2002) A genealogy of Fd-catalyzcd cross-coupling. / Organomet. Chem. 653,34-40

10. Russell SM, Tayebi N, akajima H, Riedy MC, Roberts J , Aman MJ, Migone TS, Noguchi M, Markert ML, Buckley RH, et al (1995) Mutation of Jak3 in a patient with SCID: essential role of Jak3 in lymphoid development. Science, 270, 797-800.

11. Sadowski I, Stone JC, and Pawson T. (1986) A noncatalytic domain conserved among cytϋplasrnic protem-tyrosine kinases modifies the kinase function and transforming activity of Fuji ami sarcoma virus F130gag-fps. Mσl Cell Bi l. 6, 396-408.

12. Smaill, J. B.; Palmer, B. D.; Rewcastle, G. W.; Denny, W. A.; McNamara, D. J.; Dobrusin, E. M.; Bridges, A. J.; Zhou, H.; Showalter, H. D. H.; Winters, R. T.; Leopold, W, R.; Fry, D. W.; Nelson, J. M.; SJinta , V.; Elliot, W. L.; Roberts, B. J.; Vincent, P. W.; Patmόre, S- J. (1 99) Tyrosine Kinase Inhibitors. 15. -(Phenylatnino)quinazoline and 4-(Phenylaj_aino)ρyridoId]pyrimicline Acrylarttides as Irreversible Inhibitors of the ATP Binding Site of the Epidermal Growth Factor Receptor/ Med. Chem., 42, 1803- 1815.

13. Smaill, J. B.; Rewcastle, G. W.; Loo, J. A.; Greis, K. D.; Chan, O. H.; Reyner, E. L.; Lipka, E.; Showalter, H. D. H.; Vincent, P. W.; Elliott, W. L.; Denny, W. A. (2000) Tyrosine Kinase Inhibitors. 17. Irreversible Inhibitors of the Epidermal Growth Factor Receptor: 4-(Phenylamino)quinazoline- and 4-(Phenylarrtino)pyrido[3,2- dlpyrrmidine-6-acrylamides Bearing Additional 5x>lubilizing Functions Med. Chem., 43„ 1380-1397.

14. Smaill, J . B.; Showalter, H. D. H.; Zhou, H.; Bridges, A. J.; McNamara, D. J.; Fry, D. W.; Nelson, J. M.; Sherwood, V.; Vincent, P. W. Roberts, B. J.; Elliott, W. L.; Denny, W. A. (2001) Tyrosine Kinase Inhibitors. 18.6-Substituted 4-AnilincκjUinazolines and 4-Anilmopyridof3,4-d]pyrimidines as Soluble, Irreversible Inhibitors of the Epidermal Growth Factor Receptor / Med. Che/tt., 44, 29-440.

15. Spiotto MT, and Chung TD. (2000) STAT3 mediates TL-ό-induced gr th inhibition in the human prostate cancer cell line LNCaP. Prostate 42, 88-98 16. Stille, J-K. (1986). The Palladium-Catalysed Cross-Coupling Reactions of Organotin Reagents with Organic Electrophiles. Angew. Chem., Int. Ed. £fc i.25, 508

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18. Wilks AF, Harpur AG, Kurban RR, Ralph SJ, Zurcher G, Ziemiecki A . (1991) Two novel protein-tyrosrne kinases, each with a second phosphotransferase-related catalytic domain, define a new class of protein kinase. Mol Cell Bio 1. 11, 2057-65.

19. Wilks AF, and Kurban RR (1988) Isolation and structural analysis of murine c-fes cDNA clones. Oncogerte 3, 289-94

2(1. Wissner, A.; Overbeek, E.; Reich, M. F.; Floyd, M. B.; Johnson, B. D.; Mamuya, N.; Rosfjord, E. C; Discafani, C; Davis, R.; Shi, X.; Rabindran, S. K.; Gruber, B. C; Ye, F.; Hallett, W. A.; Nilakantan, R.; Shen, R.; Wang, Y.-F.; Greenberger, L. M.; and Tsou, H.-R. (2003) Synthesis and Structure- Activity Relationships of 6,7-Disubstituted 4- Animαc uinoKne-3-carbonitriles. The Design of an Orally Active, Irreversible Inhibitor of the Tyrosine Kinase Activity of the Epidermal Growth Factor Receptor (EGFR) and the Human Epidermal Growth Factor Receptor-2 (HER-2)/ Med. Che . 46, 49-63.

Claims

1. A compound of the general formula I

or pharmaceutically acceptable prodrugs, salts, hydrates, solvates, crystal forms or diastereomers thereof, wherein: Xi, X₂, X₃, ₄ are each carbon where one is substituted with Z and the rest independently with Y; or one of X_, X₂, X₃, X₄ is N, and the others are carbon where one carbon is substituted with Z and the rest independently with Y; Λ is a ring selected from:

where D is selected from H, . alkyl, halogen, a ino; Q is a bond, halogen, C, _Λ alkyl, O, S, SO, SOj, CO, CS; Wis: (ft) NR1R2 where RI and R2 are independently H, C_M alkyl, C_M alkylCFj, aryl, hetaryl, C₁-₄ alkylaryl, Ci-t alkylhetaryl, C^ cycloalkyl, C₂-_≤ alkenyl, cyclohetalkyl, - alkylcycloalkyl, Q_i alkyl cyclohetalkyl, or RI and R2 are joined to form an optionally substituted 3-8 membered ring optionally containing an atom selected from O, S, NR3; and R3 is selected from H, -₄ alkyl, aryl, hetaryl, -₄ alkyl aryl, -₄ alkyl hetaryl, COR4 where R4 is selected from H, . alkyl, aryl, hetaryl; OR

(ii) H, _ alkyl, aryl, hetaryl, -n cycloalkyl, cyclohetalkyl, C_M alkylaryl, Cι-ι alkylhetaryl, C₃.₈ cycloalkyl, C_M alkylcycloalkyl, C alkyl cyclohetalkyl;

Y is H, halogen, CN, CF_S, nitro, OH, Cw alkyl, Q alkylNR5R6, C, ., alkylhetaryl, OQ- ₄a l, OQ-₄ al1cylOC alkyl, CC_wal riIs 5R6, C :_l.4 -dkylhetaryl, C^^ alkylcyclohetalkyl, SC_M alkyl SC2-4 alkylOC_Malkyl, SQ-4 alkylNR5R6, NR5R6, NR5COR6, NR5S0₂R6; and R5 and R6 are each independently H, -₄ alkyl, or may be joined to f rm an optionally substituted 3-6 membered ring optionally containing an atom selected from O, S, NR7 and R7 is selected from H, Ci-t alkyl, aryl, hetaryl, Q_₄ alkylaryl, Q-j alkylhetaryl; Z is selected fro :

where R8 is selected from H, Q-4 alkyl; R9 and RIO are independently selected from II, Q-4 alkyl, C_walkylNR12R13_/ Q. 4 alkylOR12, -₄ alkylhetaryl or may bo joined to for a 5-8 membered ting optionally containing an atom selected from O, S, SO, SO_z, NR14; Rll is selected from OH, OQ₄ alkyl, NR12R13; n is 0-4; where R12 and R13 are independently selected from FK Q-₄ alkyl, or may be joined to form an optionally substituted 3-8 membered ring optionally containing an atom selected from , S, NR14; and R14 is selected from H, ^ alkyl.

2. A compound according to clai 1 wherein the compound of formula I is a compound of formula Ii:

II or pharmaceutically acceptable prodrugs, salts, hydrates, solvates, crystal forms or diastereomers thereof, wherein: Xi, X?/ *, X. are each carbon where one is substituted with Z and the rest independently with Y; or one of i, Xj, X₃, X, is N, and the others are carbon where one carbon is substituted with Z and the rest independently with Y; A is a ring selected from:

where D is selected from H, .₄ alkyl, halogen, amino; Q is a bond, halogen, C alkyl, O, S, SO, SO₂, CO, CS; W is: (ii) NR1R2 where Rϊ and R2 are independently H, Q „ alkyl, Q- alkylCF_: aryl, hetaryl, Q-₄ alkylaryl, Q-, alkylhetaryl, Qv_t, cycloalkyl, _s alkenyl, cyclohetalkyl, Q-₄ alkylcycloalkyl, Q., alkyl cyclohetalkyl, or RI and R2 are joined to form an optionally substituted 3-8 membered ring optionally containing an atom selected from , S, NR3; and R3 is selected from H, CH alkyl, aryl, hetaryl, Q- alkyl aryl, Q-j alkyl hetaryl, COR4 where R4 is selected from H, Q.₄ alkyl, aryl, hetaryl; OR (ϋ) W is H, Q-₄ alkyl, aiyl, hetaryl, -g cycloalkyl, cycl ohetalkyl, Q_j alkylaryl, _₄ alkylhetaryl, _ø cycloalkyl, Cw alkylcycloalkyl, Q- alkyl cyclohetalkyl; Y is H, halogen, CN, CF,, nitro, OH, Q ₄ alkyl, C_w alkylNR5R6, C_M alkylhetaryl, OQ- * alkyl, OC₂._Λ alkylOQ-jalkyl, OQ-₄ alkylNR5R6, OCμ alkylhetaryl, OQ-* alkylcy ohetalkyl, SQ-₄ alkyl, S -4 alkylOCwalkyl, SQ-4 alkylNR5R6, NR5.R6, NR5COR6, NR5SO.R6; and R5 and R6 are each independently H, Q- alkyl, or may be joined to form an optionally substituted 3-6 membered ring optionally containing an atom selected from O, S, NR7 and R7 is selected from H, Q- alkyl, aryl, hetaryl, Q_4 alkylaryl, C« alkylhetaryl; Z is selected from :

where R8 is selected from H, Q. alkyl; R9 and RI 0 are independently selected from H, Q-₄ alkyl, Q-₄ alkylNRI2Rl3, Q. ₄ alkylOR12, Q-₄ alkylhetaryl or may be joined to form a 5-8 membered ring optionally containing an atom selected from O, S, SO, SO., NR14; Rll is selected from OH, OCu alkyl, NR12R13; n is 0-4; where: R12 and R13 arc independently selected from II, Q.₄ alkyl, or may be joined to form an optionally substituted 3-8 membered ring optionally containing an atom selected from O, S, NR14; and R14 is selected from H, Cw alkyl.

. A compound according to claim 1 selected from the group consisting of:

4. A compound according to any one of claims 1 to 3 wherein the compound irreversibly inhibits JAK-3.

5. A compound according to any one of claims 1 to 4 wherein the compound selectively inhibits JAK 3 with respect to JAK 1 or JAK 2.

6. A composition comprising a carrier and at least one compound according to any one of claims 1 to 5.

7. A method of treating a tyrosine kinase-assodated disease state, the method comprising adirunistering a therapeutically effective amount of at least one compound according to any one of claims 1 to 5 or a therapeutically effective amount of a composition according to claim 6.

8. Use of the compound according to any one of claims 1 to 5 or a composition according to claim 6 in the preparation of a medicament for the treatment of a JAK3-associated disease state.

9. A method of suppressing the immune system of a subject, the method comprising administering a therapeutically effective amount of at least one compound according to any one of claims 1 to 5 or a therapeutically effective amount of a composition according to claim 6.