Title of Invention

KINASE INHIBITOR COMPOUNDS

Abstract A compound of the general formula (I) or pharmaceutically acceptable salts, hydrates, solvates, crystal forms or diastereomers thereof is described. A method of treating protein kinase-associated disease states using the compound of formula (I) is also described. (FIG. -
Full Text KINASE INHIBITOR compounds
FIELD OF THE INVENTION
The present invention relates to the field of inhibitors of protein kinases.
BACKGROUND OF THE INVENTION
Protein kinases are a family of enzymes that catalyse the phosphorylation of
specific residues in proteins. In general protein kinases fall into several groups; those
which preferentially phosphorylate 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, including 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 cycle control and
cell growth, differentiation, apoptosis, cell mobility and mitogenesis.
Protein kinases include, for example, but are not limited to, members of the Protein
Tyrosine Kinase family (PTKs), which in turn can be divided into the cytoplasmic PTKs
and the receptor PTKs (RTKs). The cytoplasmic PTKS include the SRC family, (including:
BLK; FGR; FYN; HCK; LCK; LYN; SRQYES and YRK); the BRK Family (including: BRK;
FRK, SAD; and SRM); the CSK family (including: CSK and CTK); the BTK family,
(including BTK; ITK; TEC; MKK2 and TXK), the Janus kinase family, (including: JAKI,
JAK2, JAK3 and Tyk2), the FAK 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 ACK2); and the Abl family (including ABL and ARG). The RTK
family includes the EGF-Receptor family (including, EGFR, HER2, HER3 and HER4); the
Insulin Receptor family (including INS-R and IGF1-R); the PDGF-Receptor family
(including PDGFRa, PDGFRp, CSF1R, KIT, FLK2); the VEGF-Receptor family
(including; FLT1, FLK1 and FLT4); the FGF-Receptor family (including FGFR1, FGFR2,
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 TIE/TEK family (including TIE and TIE2/TEK); the
EPH family (including EPHA1, EPHA2, EPHA3, EPHA4, 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 SuRTK106).
Similarly, the serine /threonine specific kinases comprise a number of distinct
sub-families, including; the extracellular signal regulated kinases, (p42/ERK2 and
p44/ERKI); c-Jun NH2-terminal kinase (JNK); cAMP-responsive element-binding protein
kinases (CREBK); cAMP-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. In all of these instances,
selective inhibition of the action of the kinase might be expected to have a beneficial effect.
Diseases where aberrant kinase activity has been implicated include: diabetes; restenosis;
atherosclerosis; fibrosis 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 (Sadovski 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-terminal to JH1 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 JAK 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.
The feature most characteristic of the JAK family of PTKs is the possession of two
kinase-related domains (JH1 and JH2) (Wilks et al, 1991). The putative PTK domain of
JAK1 (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 JAK1 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
a interesting idiosyncrasy within the highly conserved sub-domain VIII 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 tryptophan
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.
The central role played by the JAK family of protein tyrosine kinases in the
cytokine dependent regulation 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
inhibitors 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
(e.g. IL-13; JAK1, JAK2), and leukemia/lymphoma (e.g. IL-2: JAK1 and JAK3).
Furthermore, certain types of cancer such as prostate cancer develop autocrine
production of certain cytokines as a selectable mechanism of developing growth and/or
metastatic potential. An example of this is cancer of the prostate, where IL-6 is produced
by and stimulates the growth of prostate cancer cell lines such as TSU and TC3 (Spiotto
MT, and Chung TD, 2000). Interestingly, levels of IL-6 are elevated in sera of patients
with metastatic prostate cancer.
A great deal of literature covers the area of cytokine signalling. The present
inventors have focussed on the JAK/STAT pathway that is involved in the direct
connection of cytokine receptor to target genes (such as cell cycle regulators (e.g. p21) and
anti-apoptosis genes (such as Bcl-XL)).
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 chain (or chains) such as the
Interleukin-4 receptor or the Interferon y receptor; (ii) a member (or members) of the JAK
family of PTKs; (iii) 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 from 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 immune 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.
SUMMARY OF THE INVENTION
The present inventors have found that a group of compounds based upon the
disubstituted pyrazine scaffold I, are inhibitors of protein kinases.
Accordingly, in a first aspect the present invention consists in a compound of the
general formula:
or pharmaceutically acceptable salts, hydrates, solvates, crystal forms or diastereomers
thereof, wherein:
D is a heterocyclic ring selected from:

where X1, X2, X3, X4 are optionally substituted carbon, or one of X1, X2, X3, X4
is N; R2 is 0-4 substituents independently chosen from H, halogen, C1-4 alkyl,
CH2F, CHF2, CF3, OCF3, aryl, hetaryl, C1-4 alkylOC1-4 alkyl, C1-4alkylOaryl,
C1-4alkylNR3R4, CO2R3, CONR3R4, CONR3SO2R4, NR3R4, C1-4alkylNR3R4,
nitro, NR3COR4, NR5CONR3R4, NR3SO2R4, C1-4alkylNR3COR4,
C1-4alkylNR5CONR3R4, C1-4alkylNR3SO2R4; and R3, R4 are each
independently H, halogen, CH2F, CHF2, CF3, C1-4 alkyl, C1-4alkyl cycloalkyl,
C1-4cydohetalkyl, aryl C1-4alkyl aryl, hetaryl, C1-4 alkyl hetaryl or may be
joined to form an optionally substituted 3-8 membered (saturated or
unsaturated) ring optionally containing an atom selected from O, S, NR6;
and R5 is selected from H, C1-4 alkyl, halogen, CH2F, CHF2, CF3, aryl or
hetaryl; and R6 is selected from H, C1-4 alkyl, aryl, hetaryl, C1-4 alkyl aryl, C1-4
alkyl hetaryl.
R1 is H, C1-4 alkyl, C1-6cycloalkyl.
Q is a bond, CH2, C1-4 alkyl
A is aryl, hetaryl optionally substituted with 0-3 substituents independently
chosen from halogen, C1-4 alkyl, CH2F, CHF2, CF3, OCF3, CN, NR8R9, aryl, hetaryl,
C1-4aryl, C1-4hetaryl, C1-4alkylNR8R9, OC1-4 alkylNR8R9, nitro, NR10C1-4NR8R9,
NR8COR9, NR10CONR8R9, NR8SO2R9, CONR8R9, CO2R8 where R8 and R9 are
each independently H, C1-4 alkyl, aryl or which together form an optionally
substituted 4-8 membered ring which may contain a heteroatom selected from O,
S, NR11, where Rll is C1-4 alkyl, and R10 is selected from H, C1-4 alkyl.
W is selected from H, C1-4alkyl, C1-4alkenyl; where C1-4alkyl or C2-6alkenyl may be
optionally substituted with C1-4alkyl, OH, OC1-4alkyl, NR12R13; and R12, and R13
are each independently H, C1-4alkyl, 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, C1-4 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.
In a third aspect the present invention consists in a method of treating a protein
kinase-associated disease state, the method comprising administering a therapeutically
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 further aspects the present invention provides the use of the compounds
described in the preparation of medicaments for the treatment of protein
kinase-associated disease states.
DETAILED DESCRIPTION
In a first aspect the present invention consists in a compound of the general formula:

or pharmaceutically acceptable salts, hydrates, solvates, crystal forms or diastereomers
thereof, wherein:
D is a heterocyclic ring selected from:

where X1, X2, X3, X4 are optionally substituted carbon, or one of X1, X2, X3, X4
is N; R2 is 0-4 substituents independently chosen from H, halogen, C1-4 alkyl,
CH2F, CHF2, CF3, OCF3, aryl, hetaryl, C1-4alkylOC1-4alkyl C1-4alkylOaryl,
C1-4alkylNR3R4, CO2R3, CONR3R4, CONR3SO2R4, NR3R4, C1-4alkylNR3R4,
nitro, NR3COR4, NR5CONR3R4, NR3SO2R4, C1-4alkylNR3COR4,
C1-4alkylNR5CONR3R4, C1-4alkyINR3SO2R4; and R3, R4 are each
independently H, halogen, CH2F, CHF2, CF3, C1-4alkyl, C1-4alkyl cycloalkyl,
C1-4 cyclohetalkyl, aryl, C1-4 alkyl aryl, hetaryl, C1-4 alkyl hetaryl, or may be
joined to form an optionally substituted 3-8 membered (saturated or
unsaturated) ring optionally containing an atom selected from O, S, NR6;
and R5 is selected from H, C1-4alkyl, halogen, CH2F, CHF2, CF3 aryl or
hetaryl; and R6 is selected from H, C1-4 alkyl, aryl, hetaryl, C1-4 alkyl aryl, C1-4
alkyl hetaryl.
R1 is H, C1-4 alkyl, C1-6cycloalkyl.
Q is a bond, CH2, C1-4 alkyl
A is aryl, hetaryl optionally substituted with 0-3 substituents independently
chosen from halogen, C1-4alkyl, CH2F, CHF2, CF3, OCF3, CN, NR8R9, aryl, hetaryl,
C1-4aryl, C1-4hetaryl, C1-4alkylNR8R9, OC1-4alkylNR8R9, nitro, NR10C1-4NR8R9,
NR8COR9, NR10CONR8R9, NR8SO2R9, CONR8R9, CO2R8 where R8 and R9 are
each independently H, C1-4 alkyl, aryl or which together form an optionally
substituted 4-8 membered ring which may contain a heteroatom selected from O,
S, NR11, where R11 is C1-4 alkyl, and R10 is selected from H, C1-4 alkyl.
W is selected from H, C1-4alkyl, C2-6alkenyl; where C1-4alkyl or C2-6alkenyl may be
optionally substituted with C1-4alkyl, OH, OC1-4alkyl, NR12R13; and R12, and R13
are each independently H, C1-4alkyl, 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, C1-4 alkyl.
In the above description it will be appreciated that:
C1-4 alkyl means a straight or branched alkyl chain
Aryl means unsubstituted or optionally substituted phenyl or naphthyl.
Hetaryl means a unsubstituted or optionally 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, NR13, where R13 is H, C1-4 alkyl, aryl, hetaryl.
In a further preferred embodiment the compound is selected from compounds of
the general formula II.
or pharmaceutically acceptable salts, hydrates, solvates, crystal forms or diastereomers
thereof, wherein:
D is a heterocyclic ring selected from:

where X1, X2, X3, X4 are optionally substituted carbon, or one of X1, X2, X3, X4
is N; R2 is 0-4 substituents independently chosen from H, halogen, C1-4 alkyl,
CH2F, CHF2, CF3, OCF3, aryl, hetaryl, C1-4alkylOC1-4 alkyl, C1-4alkylOaryl,
C1-4alkylNR3R4, CO2R3, CONR3R4, CONR3SO2R4, nitro, NR3R4, C1-4
alkylNR3R4, NR3COR4, NR5CONR3R4, NR3SO2R4, C1-4alkylNR3COR4,
C1-4alkylNR5CONR3R4, C1-4alkylNR3SO2R4; and R3, R4 are each
independently H, halogen, CH2F, CHF2, CF3, C1-4 alkyl, C1-4alkyl cycloalkyl,
C1-4 cyclohetalkyl, aryl, C1-4 alkyl aryl, hetaryl, C1-4 alkyl hetaryl, or may be
joined to form an optionally substituted 3-8 membered (saturated or
unsaturated) ring optionally containing an atom selected from O, S, NR6;
and R5 is selected from H, C1-4 alkyl, halogen, CH2F, CHF2, CF3, aryl or
hetaryl; and R6 is selected from H, C1-4 alkyl, aryl, hetaryl, C1-4 alkyl aryl, C1-4
alkyl hetaryl.
R1 is H, C1-4 alkyl, C1-4 cycloalkyl.
W is H, C1-4alkyl
A is aryl, hetaryl optionally substituted with 0-3 substituents independently
chosen from halogen, C1-4 alkyl, CH2F, CHF2, CF3, OCF3, CN, nitro, NR8R9, aryl,
hetaryl, C1-4aryl, C1-4hetaryl, C1-4 alkylNR8R9, OC1-4alkylNR8R9, NR10C1-4NR8R9,
NR8COR9, NR10CONR8R9, NR8SO2R9, CONR8R9, CO2R8 where R8 and R9 are
each independently H, C1-4 alkyl, aryl or which together form an optionally
substituted 4-8 membered ring which may contain a heteroatom selected from O,
S, NR11, where R11 is C1-4 alkyl, and R10 is selected from H, C1-4 alkyl.
In the above description it will be appreciated that:
C1-4 alkyl means a straight or branched alkyl chain
Aryl means unsubstituted or optionally substituted phenyl or naphthyl.
Hetaryl means a unsubstituted or optionally substituted 5- or 6-membered
heteroaromatic ring containing one or more heteroatoms selected from O, N, S.
Cydoalkyl means a 3-8 membered saturated ring
Cyclohetalkyl means a 3-8 membered saturated ring containing 1-3 heteroatoms
selected from O, S, NR13, where R13 is H, C1-4 alkyl, aryl, hetaryl.
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 all optical isomers and stereoisomers of the compounds of the present
invention, and mixtures thereof, and to all pharmaceutical compositions and methods of
treatment that may employ or contain them. In this regard, the invention includes both
the E and Z configurations. The compounds of formula I may also exist as tautomers.
This invention relates to the use of all such tautomers 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 carboxylic
groups can be converted into prodrugs. Prodrugs include compounds wherein an amino
acid residue, or a polypeptide chain of two or more (eg, two, three or four) amino acid
residues which are covalently joined through pepnde bonds to free amino, hydroxy and
carboxylic acid groups of compounds of formula I. The amino acid residues include the
20 naturally occurring amino acids commonly designated by three letter symbols and also
include, 4-hydroxyproline, hydroxylysine, demosine, isodemosine, 3-methylhisndine,
norvlin, beta-alanine, gamma-aminobutyric acid, citrulline, homocysteine, homoserine,
ornithine and methioine sulfone. Prodrugs also include compounds wherein carbonates,
carbamates, amides and alkyl esters which are covalently bonded to the above
substituents of formula I 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 bond to a free hydroxyl of compounds of
formula I.
In a still further preferred embodiment the compound possesses S chirality at the
chiral carbon bearing W, where W is C1-4 alkyl. 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 still further preferred embodiment the compound is selected from the group
consisting of 6-(1H-benzimidazol-1-yl)-N-[(1R)-1-phenylethyl]pyrazin-2-amine, N-benzyl-
6-(1H-imidazol-1-yl)pyrazin-2-amine, 6-(1H-benzimidazol-1-yl)-N-[(15)-1-(4-
methoxyphenyl)ethyl]pyrazin-2-amine, N-(4-Fluorobenzyl)-6-(1H-imidazol-1-yl)pyrazin-
2-amine, 6-(1H-benzimidazol-1-yl)-N-[(1S)-1-(4-bromophenyl)ethyl]pyrazin-2-amine,
6-(1H-Imidazol-1-yl)-N-[(1R)-1-phenylethyl]pyrazin-2-amine, 1-(6-{[(15)-1-
phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazole-6-carboxamide, 6-(1H-Benzimidazol-
1-yl)-N-benzylpyrazin-2-amine, 1-(6-{[(1S)-1-phenylethyl]amino)pyrazin-2-yl)-1H-
benzimidazole-5-carboxamide, 6-(1H-Benzimidazol-1-yl)-N-(4-fluorobenzyl)pyrazin-2-
amine, 6-{5-[(Morpholino-1-yl)carbonyl]-1H-benzimidazol-1-yl}-N-[(1S)-1-
phenylethyl]pyrazin-2-amine, 6-(1H-imidazo[4,5-b]pyridin-1-yl)-N-[(1R)-1-
phenylethyl]pyrazin-2-amine, 6-{6-[(Morpholino-1-yl)carbonyl]-1H-benzimidazol-1-yl}-N-
[(1S)-1-phenylethyl]pyrazin-2-amine, 6-(1H-imidazol-1-yl)-N-(4-morpholin-4-
ylphenyl)pyrazin-2-amine, N-[1-(6-{[(1S)-1-Phenylethyl]amino}pyrazin-2-yl)-1H-
benzimidazol-6-yl]cyclopropanecarboxamide, N-[1-(6-{[((1S)-1-
phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-5-yl]nicotinamide, N-[1-(6-{[(1S)-1-
Phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-5-yl]cydopropanecarboxamide,
6-(1H-Benzimidazol-1-yl)-N-[(1R)-1-phenylethyl]pyrazin-2-amine, 6-[6-(4,5-dihydro-1,3-
oxazol-2-yl)-1H-benzimidazol-1-yl]-N-[(1S)-1-phenylethyl]pyrazin-2-amine, N-[(1R)-1-
Phenylethyl]-6-(4-phenyl-1H-imidazol-1-yl)pyrazin-2-amine, 1-[6-{[(1S)-1-
phenylethyl]amino}pyrazin-2-yl]-N-(2-hydroxyethyl)-1H-benzimidazole-6-carboxamide,
N-[1-(6-{[(15)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-6-
yl]methanesulfonamide, N-[l-(6-{[(15)-1-phenylethyl]amino}pyrazin-2-yl)-1H-
benzimidazol-5-yl]methanesulfonamide, N-[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-
yl)-1H-benzimidazol-5-yl]isonicotinamide, 6-(1H-Imidazol-1-yl)-N-[(1S)-1-
phenylethyl]pyrazin-2-amine, N-[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-
benzimidazol-6-yl]isonicotinamide, 6-(1H-BenzimidazoI-1-yl)-N-[(1S)-1-
phenylethyl]pyrazin-2-amine, 6-[5-(4,5-dihydro-1,3-oxazol-2-yl)-1H-benzimidazol-1-yl]-
N-[(1S)-1-phenylethyl]pyrazin-2-amine, 1-[6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl]-N-
(2-hydroxyethyl)-1H-benzimidazole-5-carboxamide, 6-(5-Methyl-1H-benzimidazol-1-yl)-
N-[(1S)-1-phenylethyl]pyrazin-2-amine,N-[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-
1H-benzimidazol-6-yl]nicotinamide, N-methyl-1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-
yl)-1H-benzimidazole-5-carboxamide, N-[1-(6-{[(1S)-1-phenylethyl]amino)pyrazin-2-yl)-
lH-benzimidazol-6-yl]-2,2-dimethylpropanamide, N-methyl-1-(6-{[(15)-1-
phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazole-6-carboxamide, N-[1-(6-{[(1S)-1-
phenylethyl]amino)pyrazin-2-yl)-1H-benzimidazol-5-yl]-2,2-dimethylpropanamide, 1-(6-
{[(1S)-1-Phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-5-amine,2-Methoxy-N-[1-(6-
{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-5-yl]acetamide, 1-(6-{[(1S)-1-
phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-6-amine, 2-Methoxy-N-[1-(6-{[(1S)-1-
phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-6-yl]acetamide, N-Benzyl-1-[6-([(1S)-
l-phenylethyl]amino)pyrazin-2-yl]-1H-benzimidazole-5-carboxamide, N-[1-(6-{[(1S)-1-
phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-5-yl]pyrazine-2-carboxamide, 1-(6-
{[(1S)-1-Phenylethyl]amino}pyrazin-2-yl)-N-phenyl-1H-benzimidazole-5-carboxamide,
N-[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-6-yl]pyrazine-2-
carboxamide,N-[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-6-
yl]acetamide, 6-{5-[(4-Methylpiperazin-1-yl)methyl]-1H-benzimidazol-1-yl}-N-[(15)-1-
phenylethyllpyrazin-2-amine, N-[1-(6-{[(1S)-1-Phenylethyllaminolpyrazin-2-yl-1H-
benzimidazol-5-yl]acetamide, [1-(6-{[(1S)-1-Phenylethyl]amino}pyrazin-2-yl)-1H-
benzimidazol-5-yl]methanol/N-[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-
benzimidazol-6-yl]benzamide, [1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-
benzimidazol-6-yl]methanol, N-[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-
benzimidazol-5-yl]benzamide, 1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-N-[2-
(dimethylamino)ethyl]-1H-benzimidazole-5-carboxamide, 1-[6-{[(1S)-1-
Phenylethyl]amino}pyrazin-2-yl]-N-(pyridin-3-ylmethyl)-1H-benzimidazol-5-amine,
tert-butyl(2S)-2-({[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-5-
yl]amino}carbonyl)pyrrolidine-1-carboxylate,6-(3H-imidazo[4,5-c]pyridin-3-yl)-N-[(1S)-1-
phenylethyl]pyrazin-2-amine, 6-(1H-benzimidazol-1-yl)-N-[1-(4-
fluorophenyl)ethyl]pyrazin-2-amine, 6-(1H-imidazo[4,5-c]pyridin-1-yl)-N-[(1S)-1-
phenylethyl]pyrazin-2-amine, 6-(1H-benzimidazol-1-yl)-N-[(l5)-1-(4-pyridin-3-
yIphenyl)ethyl]pyrazin-2-amine, (2S)-N-[1-(6-{[(1S)-1-phenylethyl]amino)pyrazin-2-yl)-
1H-benzimidazol-5-yl]pyrroIidine-2-carboxamide/N-[(1S)-1-phenylethyl]-6-(5-pyridin-4-
yl-1H-benzimidazol-1-yl)pyrazin-2-amine, N-[(15)-1-phenylethyl]-6-(5-pyridin-3-yl-1H-
benzimidazol-1-yl)pyrazin-2-amine, 6-(5-bromo-1H-benzimidazol-1-yl)-N-[(1S)-1-
phenylethyl]pyrazin-2-amine, N-[3-(1H-imidazol-1-yl)propyl]-1-[6-([(1S)-1-
phenylethyl]amino)pyrazin-2-yl]-1H-benzimidazole-6-carboxamide, N-1H-
benzimidazole-6-carboxamide,N-(3-morpholin-4-yIpropyl)-1-[6-([(1S)-1-
phenylethyl]amino)pyrazin-2-yl]-1H-benzimidazole-6-carboxamide, N-(3-morpholin-4-
ylpropyl)-1-[6-([(1S)-1-phenylethyl]amino)pyrazin-2-yl]-1H-benzimidazole-5-
carboxamide, N-[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-5-
yl]piperidine-3-carboxamide, 6-(1H-benzimidazol-1-yl)-N-[(1S)-1-pyridin-3-
yIethyl]pyrazin-2-amine,6-(1H-benzimidazol-1-yl)-N-[(1S)-1-(1,1"-biphenyl-4-
yl)ethyl]pyrazin-2-amineN-[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-
benzimidazol-5-yl]benzenesulfonamideand 6-(1H-benzimidazol-1-yl)-N-[(1S)-1-(1,1"-
biphenyl-4-yl)ethyl]pyrazin-2-amine.
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 protein
kinase-associated disease state, the method comprising administering a therapeutically
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 preferred embodiment, the disease state involves a receptor tyrosine kinase
selected from the group consisting of EGF, HER2, HER3, HER4, IR, IGF-1R, IRR,
PDGFR.alpha., PDGFR.beta., CSFIR, C-Kit, C-fms,Flk-lR, Flk4, KDR/Flk-1, Flt-1, FGFR-
1R, FGFR-2R, FGFR-3R and FGFR-4R.
In another preferred embodiment, the disease state involves a cellular tyrosine
kinase selected from the group consisting of Src, Frk, Btk, Csk, Abl, ZAP70, Fes/Fps, Fak,
Ack, Yes, Fyn, Lyn, Lck, Blk, Hck, Fgr and Yrk.
In a further preferred embodiment, the disease state involves a tyrosine kinase
selected from the group consisting of JAK1, JAK2, JAK3 and TYK2.
In a yet further preferred embodiment, the disease state involves a
serine/threonine kinase selected from the group consisting of ERK2, c-Jun, p38 MAPK,
PKA, PKB, PKC, a cydin-dependent kinase, CDK1, CDK2, CDK3, CDK4, CDK5, CDK6,
CDK7, CDK8, CDK9, CDK10, and CDK11.
In a preferred embodiment of the present invention the disease state is selected
from the group consisting of Atopy, such as Allergic Asthma, Atopic Dermatitis (Eczema),
and Allergic Rhinitis; Cell Mediated Hypersensitivity, such as Allergic Contact
Dermatitis and Hypersensitivity Pneumonitis; Rheumatic Diseases, such as Systemic
Lupus Erythematosus (SLE), Rheumatoid Arthritis, Juvenile Arthritis, Sjogren"s
Syndrome, Sderoderma, Polymyositis, Ankylosing Spondylitis, 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, Varicella-Zoster Virus (VZV), Human Papilloma Virus (HPV),
Cancer, such as Leukemia, Lymphoma and Prostate Cancer.
As used herein the term "protein kinase-associated disease state" refers to those
disorders which result from aberrant protein kinase activity, in particular JAK activity
and / or which are alleviated by inhibition of one or more of these enzymes.
In further aspects the present invention provides the use of the compounds
described in the preparation of medicaments for the treatment of protein
kinase-associated disease states.
As used herein the term "JAK", "JAK kinase" or "JAK family" refers to protein
tyrosine kinases which possess the characterizing features of JAK1, JAK2, JAK3 and TYK
as described herein.
The present invention provides pharmaceutical compositions comprising at least
one of the compounds of the present invention capable of treating a protein
kinase-associated disorder, such as a JAK associated disorder, in an amount effective
therefor, and a pharmaceutically acceptable vehicle or diluent. 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 well as pharmaceutical additives of a type appropriate to the mode of desired
administration (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 present invention may be administered by any suitable
means, for example, orally, such as in the form of tablets, capsules, granules or powders;
sublingually; buccally; parenterally, such as by subcutaneous, intravenous, intramuscular,
or intracisternal injection or infusion techniques (e.g., as sterile injectable aqueous or
non-aqueous solutions or suspensions); nasally such as by inhalation 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
diluents. 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. The compounds may also be administered
liposomally.
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 inflammation 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 inflammatory response.
The subjects treated in the above methods, in whom which JAK inhibition is
desired, are 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,
and preferably a human being, male or female.
The term "therapeutically 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 "administering 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 well known in the art of pharmacy. All 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 liquid
carrier or a finely divided solid 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
well as any product which results, directly or indirectly, 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 pharmaceutically acceptable excipients which are
suitable for the manufacture of tablets. These excipients may be for example, inert
diluents, 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 glyceryl 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 carboxymethylcellulose, methylcellulose,
hydroxy-propylmethylcellulose, sodium alginate, polyvinyl-pyrrolidone, gum tragacanth
and gum acacia; dispersing or wetting agents may be a naturally-occurring phosphatide,
for example lecithin, 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.
Oily suspensions may be formulated by suspending the active ingredient in a
vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in a mineral
oil such as liquid paraffin. The oily 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 exemplified 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-water emulsions. The oily phase may be a vegetable oil, for example olive oil or
arachis oil, or a mineral oil, for example liquid paraffin or mixtures of these. Suitable
emulsifying agents may be naturally- occurring gums, for example gum acacia or gum
tragacanth, naturally-occurring phosphatides, for example soy bean, lecithin, 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 parenterally-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 acid find use in the
preparation of intertables.
The compounds of the present invention may also be administered in the form of
suppositories for rectal administration of the drug. These compositions can be prepared
by mixing the drug with a suitable non-irritating excipient which is solid 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 application shall include mouthwashes and gargles.)
The compounds of the present invention can also be administered in the form of
liposomes. As is known in the art, liposomes are generally derived from phospholipids or
other lipid substances. Liposomes are formed by mono- or mululamellar hydrated liquid
crystals that are dispersed in an aqueous medium. Any non-toxic, physiologically
acceptable and metabolisable lipid capable of forming liposomes can be used. The
present compositions in liposome form can contain, in addition to a compound of the
present invention, stabilisers, preservatives, excipients and the like. The preferred lipids
are the phospholipids 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 therapeutically active compounds as noted herein which are usually
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 synergistically 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-IL-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
specific for CD40 and/or gp39 (i.e., CD154), fusion proteins constructed from CD40 and
gp39 (CD401g and CD8gp39), inhibitors, such as nuclear translocation inhibitors, of
NF-kappa B function, such as deoxyspergualin (DSG), cholesterol biosynthesis inhibitors
such as HMG CoA reductase inhibitors (lovastatin and simvastatin), non-steroidal
antiinflammatory drugs (NSAIDs) such as ibuprofen, aspirin, acetaminophen and
cyclooxygenase inhibitors such as rofecoxib, steroids such as prednisolone or
dexamethasone, gold compounds, antiproliferative agents such as methotrexate, FK506
(tacrolimus, Prograf), mycophenolate mofetil, cytotoxic drugs such as azathioprine, VP-
16, etoposide, fludarabine, cisplatin and cyclophosphamide, TNF-a 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.
In 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 will be about 0.1 to about 250 mg/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 administration, the compositions are preferably provided in the form of tablets
containing 1.0 to 1000 milligrams of the active ingredient, particularly 1.0, 5.0, 10.0, 15.0.
20.0, 25.0, 50.0, 75.0, 100.0, 150.0, 200.0, 250.0, 300.0, 400.0, 500.0, 600.0, 750.0, 800.0, 900.0,
and 1000.0 milligrams of the active ingredient for the symptomatic adjustment of the
dosage to the patient to be treated. The compounds may 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 will depend upon a variety of factors
including the activity of the specific compound employed, the metabolic stability 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.
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 before the priority date of each
claim of this application.
In order that the nature of the present invention may be more clearly understood
preferred forms thereof will now be described by reference to the following non-limiting
Examples.
MATERIALS AND METHODS:
Compound Synthesis
Compounds are generally prepared in a 2-step process starting from
2,6-dichloropyrazine.
The first step is a nudeophilic aromatic substitution to generate a
monoamino-monohalo intermediate. (Scheme 1).

The nudeophilic aromatic substitution is typically carried out by addition of a
primary amine to the di-halogenated heterocyde in a solvent such as ethanol,
isopropanol, tert-butanol, dioxane, THF, DMF, toluene or xylene. The reaction is typically
performed at elevated temperature in the presence of excess amine or a non-nucleophilic
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, Pd2(dba)3/BINAP and Pd(OAc)2/BINAP.
The amines employed in the first step of the synthesis of these compounds are
obtained commercially or are prepared using methods well known to those skilled in the
art. Of particular interest are a-methylbenzylamines which are obtained commercially or
may be prepared through reduction of oximes (Scheme 2). Typical reductants include
lithium aluminium hydride, hydrogen gas in the presence of catalytic palladium on
charcoal, Zn in the presence of hydrochloric acid, sodium borohydride in the presence of
a Lewis acid such as TiCl3, ZrCl4, NiCl2 and MoO3 or sodium borohydride in conjunction
with Amberlyst H15 ion exchange resin and LiCl. The oximes are obtained in one-step
from the corresponding ketones through condensation with hydroxylamine. This
reaction is generally performed in a protic solvent such as water or ethanol, at
temperatures from 0°C to reflux. The hydroxylamine is generally used in the form of its
hydrochloride salt, and therefore the reaction is performed in the presence of a base such
as sodium hydroxide. The ketones employed as starting materials are generally obtained
commercially or via procedures well known to those skilled in the art.

a-Methyl benzylamines of high optical purity may be prepared from chiral
a-methyl benzyl alcohols using methods well known to those skilled in the art. Such
methods include derivatisation of the hydroxyl as a mesylate or tosylate and
displacement with a nitrogen nudeophile, such as phthalimide or azide which can then
converted to the primary amine using conventional synthetic methods; or, displacement
of the hydroxyl with a suitable nitrogen nucleophile under Mitsunobu conditions. The
chiral a-methyl benzyl alcohols may be obtained through chiral reduction of the
corresponding ketones. Chiral reducing methods are now well known in organic
chemistry and include enzymatic processes, asymmetric hydrogenation procedures and
chiral oxazaborolidines.
The second step of the synthesis involves a nucleophilic aromatic substitution
reaction of the monochloro- mono-amino pyrazine with imidazole, benzimidazole or
indazole. The reaction is typically performed using a salt of the imidazole, benzimidazole
or indazole in solvents such as tetrahydrofuran, dimethylformamide, toluene, or xylene
from room temperature to reflux. The imidazole, benzimidazole or indazole 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 imidazole, benzimidazole or indazole ring. The reaction is
typically performed using a base such as caesium carbonate, rubidium carbonate,
potassium carbonate, sodium tert-butoxide or potassium phosphate in a solvent such as
xylene, toluene, and DMF from room temperature to reflux. Auxiliary reagents such as
phase transfer agents (e.g. cetrimonium bromide) or copper complexing agents (e.g.
phenanthroline) may also be employed in the reaction.
The imidazole, benzimidazole or indazole components used in this reaction are
obtained commercially or are prepared from commercially available imidazoles,
benzimidazoles or indazoles via techniques well known to those skilled in the art.
Alternatively, an imidazole, benzimidazole or indazole derivative may be reacted
with the mono-amino mono-chloro pyrazine and the subsequent product further
derivatised using methods well known to those skilled in the art.
Representative syntheses are reported below.
Example 1
6-Chloro-N-[(1R)-1-phenylethy]]pyrazin-2-amine

A solution of R-a-methylbenzylamine (3.64g, 30.0mmol) and 2,6-dichloropyrazine
(1.50g, 10.0mmol) in dioxane (5 mL) was heated at reflux under N2 for 48 hours. The
solvent was removed and the product crystallised from toluene-hexane.
1H-n.m.r. (CDCl3) d1.59 (d, 3H, J=6.9Hz, CH3), 4.88 (q, 1H, J=6.6Hz, CH), 5.13 (br s,
1H, NH), 7.27-7.36 (m, 5H, ArH), 7.64 (s, 1H, pyraz-H), 7.79 (s, 1H, pyraz-H).
Example 2
6-(1H-Imidazol-1-yl)-N-[(1R)-1-phenylethyl]pyrazin-2-amine

To a stirred solution of imidazole (82mg, 1.2mmol) in anhydrous DMF (5mL) at
0°C under N2 was added sodium hydride (58mg, 60% dispersion in oil, 1.45mmol). The
mixture was stirred at 0°C for 10 min and at RT for 30 min. To this mixture was added a
solution of 6-chloro-N-[(1R-1-phenylethyl]pyrazin-2-amine (280mg, 1.2mmol) in DMF
(5mL). The mixture was then stirred at RT for 62h followed by heating at reflux for 18h.
The DMF was removed under reduced pressure and the residue diluted with chloroform.
The organic layer was washed with water, dried (Na2SO4) and the solvent removed under
reduced pressure to furnish the crude product. Column chromatography using
dichloromethane-methanol (19:1 ? 10:1) as eluant separated recovered starting material
from product (177mg, 56%).
1H-n.m.r. (CDCl3) d1.62 (d, 3H, J = 6.9Hz, CH3), 4.97 (m, 1H, CH), 5.46 (d, 1H, J =
6.0Hz, NH), 7.17 (s, 1H, imid-H), 7.23-7.40 (m, 5H, Ph-H), 7.47 (s, 1H, imid-H), 7.76 (s, 1H,
pyraz-H), 7.91 (s, 1H, pyraz-H), 8.20 (s, 1H, imid-H).
m/z (ES) 266 (M++H), 162,105.
Example 3
6-Chloro-N-(4-morpholin-4-ylphenyl)pyrazin-2-amine

In a procedure analogous to example 1, reaction of 4-morpholinoaniline (2.15g,
12.1mmol) and 2,6-dichloropyrazine (0.756g, 5.03mmol) furnished the product (0.54g,
37%).
1H-n.m.r. (CDCl3)d 3.25 (br s, 4H, CH2), 3.99 (br s, 4H, CH2), 7.05-7.17 (m, 2H,
ArH), 7.42-7.54 (m, 2H, ArH), 7.94 (s, 1H, pyraz-H), 8.04 (s, 1H, pyraz-H), 8.06 (s, 1H,
NH).
Example 4
6-Chloro-N-(4-fluorobenzyl)pyrazin-2-amine

In a procedure analogous to example 1, reaction of 4-fluorobenzylamine (3.75 g,
30.0mmol) and 2,6-dichloropyrazine (1.49 g, 10.0mmol) furnished the product (2.35 g,
99%).
1H-n.m-r. (CDCl3) 54.53(s,2H, CH2),5.08(br s, 1H, NH), 7.01-7.07(m, 2H, ArH),
7.30-7.34(m,2H, ArH), 7.77(s, 1H, pyraz-H), 7.83(s, 1H, pyraz-H)
Example 5
6-(1H-Imidazol-1-yl)-N-(4-morpholin-4-ylphenyl)pyrazin-2-amine

In a procedure analogous to example 2, reaction of
6-chloro-N-(4-morpholin-4-ylphenyl)pyrazin-2-amine (100mg, 0.34mmol) and imidazole
(126mg, 0.38mmol) furnished the product (37mg, 34%).
1H-n.m.r. (CDCl3) d 3.18 (br s, 4H, 2CH2), 3.87-3.91 (m, 4H, 2CH2), 6.77 (s, 1H,
imid-H), 6.94-6.98 (m, 2H, ArH), 7.36 (d, 2H, J =8.7Hz, ArH), 7.62 (br s, 1H, imid-H), 8.03
(s, 1H, pyraz-H), 8.08 (s, 1H, pyraz-H), 8.39 (br s, 1H, imid-H).
m/z (ES) 323 (M++H).
Example 6
N-(4-Fluorobenzyl)-6-(1H-imidazol-1-yl)pyrazin-2-amine

In a procedure analogous to example 2, reaction of
6-chloro-N-(4-fluorobenzyl)pyrazin-2-amine (240mg, 1.01mmol) and imidazole (76mg,
1.11mmol) furnished the product (210mg, 65%).
1H-n.m.r. (CDCl3) 8 4.59 (d, 2H, J=5.7Hz, CH2), 5.23 (t-like, 1H, NH), 7.03-7.08 (m,
2H, ArH), 7.20 (s, 1H, imid-H), 7.32-7.37 (m, 2H, ArH), 7.55 (s, 1H, imid-H), 7.85 (s, 1H,
pyraz-H), 8.00 (s, 1H, pyraz-H), 8.29 (s, 1H, imid-H).
m/z(ES)270(M++H).
Example 7
6-(2-Methyl-1H-imidazol-1-yl)-N-[(1R)-1-phenylethyl]pyrazin-2-amine

In a procedure analogous to example 2, reaction of
6-chloro-N-[(1R)-1-phenylethyl]pyrazin-2-amine (150mg, 0.64mmol) and
2-methylimidazole (58mg, 0.71mmol) furnished the product (172mg, 40%).
1H-n.m.r. (CDCl3) d1.59 (d, 3H, J = 6.8Hz, CH3), 2.43 (s, 3H, CH3), 4.98 (m, 1H, CH),
5.45 (br s, 1H, NH), 6.98 (d, 1H, J= 1.3 Hz, imid-H), 7.17 (d, 1H, J = 1.3 Hz, imid-H),
7.22-7.35 (m, 5H, ArH), 7.82 (s, 1H, pyraz-H), 7.84 (s, 1H, pyraz-H).
m/z(ES)280(M++H).
Example 8
M-(4-Fluorobenzyl)-6-(2-methyl-1H-imidazol-1-y])pyrazin-2-amine

In a procedure analogous to example 2, reaction of
6-chloro-N-(4-fluorobenzyl)pyrazin-2-amine (150mg, 0.63mmol) and 2-methylimidazole
(57mg, 0.69mmol) furnished the product (42mg, 23%).
1H-n.m.r. (CDCl3) d2.56 (s, 3H, CH3), 4.57 (d, J= 5.7 Hz, 1H, CH2), 5.34 (br s, 1H,
NH), 7.01-7.07 (m, 3H, Ar-H), 7.26 (s, 1H, imid-H), 7.29-7.34 (m, 2H, ArH), 7.92 (s, 2H,
pyraz-H).
m/z(ES)284(M++H).
Example 9
N-(4-Fluorobenzyl)-6-(4-methyl-1H-imidazol-1-yl)pyrazin-2-amine and
N-(4-fluorobenzyl)-6-(5-methyl-1H-imidazol-1-yl)pyrazin-2-amine

In a procedure analogous to example 2, reaction of
2-(4-fluorobenzylamino)-6-chloro-pyrazine (190mg, 0.80mmol) and 4-methylimidazole
(72mg, 0.88mmol) furnished the following products: 4-methyl derivative (100mg, 44%);
5-methyl derivative (19mg, 8%).
(4-methyl derivative) 1H-n.m.r. (CDCl3) d 2.31 (s, 3H, CH3), 4.58 (d, 2H, J = 5.7 Hz,
CH2), 5.40 (br s, 1H, NH), 6.92 (s, 1H, imid-H), 7.00-7.08 (m, 2H, ArH), 7.25 (s, 1H,
imid-H5), 7.31-7.36 (m, 2H, ArH), 7.84 (s, 1H, pyraz-H5), 7.93 (s, 1H, pyraz-H3), 8.24 (br s,
1H, imid-H2).
m/z(ES)284(M++H).
(5-methyl derivative) 1H-n.m.r. (CDCl3) d 2.34 (s, 3H, CH3), 4.57 (d, 2H, J= 5.7 Hz,
CH2), 5.44 (br s, 1H, NH), 6.92 (s, 1H, imid-H), 7.00-7.07 (m, 2H, ArH), 7.28-7.34 (m, 2H,
ArH), 7.93 (s, 1H, pyraz-H), 7.95 (s, 1H, pyraz-H), 7.98 (br s, 1H, imid-H).
m/z (ES) 284 (M++H).
Example 10
N-[(1R)-1-Phenylethyl]-6-(4-phenyl-1H-imidazol-1-yl)pyrazin-2-amine

In a procedure analogous to example 2, reaction of
6-chloro-N-[(1R)-1-phenylethyl]pyrazin-2-amine (180mg, 0.77mmol) and
4-phenylimidazole (122mg, 0.85mmol) furnished the product (176mg, 67%).
1H-n.m.r. (CDCl3) d1.63 (d, 3H, J = 6.9Hz, CH3), 4.93-5.02 (m, 1H, CH), 5.26 (d, 1H,
J = 6.0Hz, NH), 7.25-7.44 (m, 8H, ArH), 7.72 (d, 1H, J= 1.2 Hz, imid-H), 7.77 (s, 1H,
pyraz-H), 7.82-7.86 (m, 2H, ArH), 7.92 (s, 1H, pyraz-H), 8.22 (s, 1H, imid-H).
m/z(ES)342(M++H).
Example 11
N-Benzyl-6-chloropyrazin-2-amine

In a procedure analogous to example 1, reaction of benzylamine (3.21g, 30.0mmol)
and 2,6-dichloropyrazine (1.49g, 10.0mmol) furnished the product ( 2.15g, 98%).
1H-n.m.r. (CDCl3) d 4.55 (d, 2H, J = 5.7Hz, CH2), 7.28-7.40 (m, 5H, ArH), 7.76 (s, 1H,
pyraz-H), 7.83 (s, 1H, pyraz-H).
Example 12
6-(1H-Benzimidazol-1-yl)-N-benzylpyrazin-2-amine

To a stirred solution of benzimidazole (130mg, 1.1mmol) in anhydrous DMF (5mL) at 0°C
under N2 was added sodium hydride (56mg, 60% dispersion in oil, 1.45mmol) in portions
over 2 min. The mixture was stirred at 0°C for 15 min and at RT for 60 min. To this was
added a solution of (6-chloro-pyrazin-2-yl)-(1-benzyl)-amine (220mg) in DMF (5mL) and
the resulting mixture was then heated at reflux for 18h. The DMF was removed under
reduced pressure and the residue diluted with chloroform. The organic layer was washed
with water, dried (Na2SO4) and the solvent removed under reduced pressure to furnish
the crude product. Column chromatography using dichloromethane-methanol (20:1 ?
10:1) as eluant separated the product (100mg).
1H-n.m.r. (CDCl3) d 4.66 (d, 2H, J = 5.7Hz, CH2), 5.56 (m, 1H, NH), 7.29-7.39 (m,
7H, Ar-H), 7.78-7.89 (m, 2H, Ar-H), 7.92 (s, 1H, pyraz-H), 8.16 (s, 1H, pyraz-H), 8.48 (s,
1H, benzimid-H2).
m/z(ES)302(M++H).
Example 13
6-(1H-Benzimidazol-1-yl)-N-[(1R)-1-phenylethyl]pyrazin-2-amine

In a procedure analogous to example 12, reaction of
6-chloro-N-[(lR)-1-phenylethyl]pyrazin-2-amine (240mg, 1.03mmol) and benzimidazole
(130mg, 1.10mmol) furnished the product (187mg, 59%).
1H-n.m.r. (CDCl3) d1.63 (d, 3H, J = 6.6Hz, CH3), 4.98-5.20 (m, 1H, CH), 5.58 (d, 1H,
J=6.0Hz, NH), 7.25-7.42 (m, 6H, Ph-H, benzimid-H), 7.70 (dd, 1H, J = 7.2, 1.0Hz,
benzimid-H), 7.82 (dd, lH,J = 8.0, 1.2Hz, benzimid-H), 7.87 (s, 1H, pyraz-H), 8.11 (s, 1H,
pyraz-H), 8.38 (s, 1H, benzimid-H).
m/z (ES) 315 (M++H), 212, 105.
Example 14
6-(1H-Benzimidazol-1-yl)-N-[(S)-1-phenylethyl]pyrazin-2-amine

In a procedure analogous to example 12, reaction of
6-chloro-N-[(1S)-1-phenylethyl]pyrazin-2-amine (140mg, 0.60mmol) and benzimidazole
(78mg, 0.66mmol) furnished the product (71mg, 38%).
1H-n.m.r. (CDCl3) d1.57 (d, 3H, J = 6.9Hz, CH3), 4.95 (m, 1H, CH), 5.29 (d, 1H,
J=6.0Hz, NH), 7.19-7.35 (m, 7H, Ph-H, benzimid-H), 7.63-7.66 (m, 1H, benzimid-H),
7.74-7.77 (m, 1H, benzimid-H), 7.78 (s, 1H, pyraz-H), 8.06 (s, 1H, pyraz-H), 8.31 (s, 1H,
benzimid-H).
m/z (ES) 316 (M++H), 212, 105
Example 15
6-(1H-Benzimidazol-1-yl)-N-(4-morpholin-4-ylphenyl)pyrazin-2-amine

In a procedure analogous to example 12, reaction of
6-chloro-N-(4-morpholin-4-ylphenyl)pyrazin-2-amine (150mg, 0.52mmol) and
benzimidazole (67mg, 0.57mmol) furnished the product (60mg, 31%).
1H-n.m.r. (CDCl3) d 3.19 (br s, 4H, 2CH2), 3.90 (t, 4H, J=4.6Hz, 2CH2), 6.69 (s, 1H,
NH), 6.98 (d, 2H, J=8.4 Hz, ArH), 7.37-7.41 (m, 4H, ArH), 7.87-7.90 (m, 1H, ArH), 8.00-8.03
(m, 1H, ArH), 8.08 (s, 1H, pyraz-H), 8.31 (s, 1H, pyraz-H), 8.59 (s, 1H, benzimid-H2).
m/z(ES)373(M++H).
Example 16
6-(1H-Benzimidazol-1-yl)-N-(4-fluorobenzyl)pyrazin-2-amine

In a procedure analogous to example 12, reaction of
6-chloro-N-(4-fluorobenzyI)pyrazin-2-amine (240mg, 1.01mmol) and benzimidazole
(130mg, 1.1mmol) furnished the product (170mg, 53%).
1H-n.m.r. (CDCl3) d4.64 (d, 2H, J = 5.7Hz, CH2), 5.46 (br s, 1H, NH), 7.06 (m, 2H,
ArH), 7.30-7.38 (m, 4H, ArH), 7.82-7.88 (m, 2H, ArH), 7.93 (s, 1H, pyraz-H), 8.20 (s, 1H,
benzimid-H), 8.49 (s, 1H, pyraz-H).
m/z(ES)320(M++H).
Example 17
6-(1H-imidazo[4,5-b]pyridin-1-yl)-N-[(1S)-1-phenylethyl]pyrazin-2-amine and
6-(3H-imidazo[4,5-b]pyridin-3-yl)-N-[(R)-1-phenylethyl]pyrazin-2-amine

In a procedure analogous to example 12, reaction of
6-chloro-N-[(1S)-1-phenylethyl]pyrazin-2-arnine (240mg, 1.03mmol) and
4-azabenzimidazole (130mg, 1.09mmol) furnished the product (7mg, 2%) as a 1:1 mixture
of regioisomers.
1H-n.m.r. (as 1:1 mixture) (CDCl3) d1.54 (d, 3H, CH3), 1.63 (d, 3H, CH3), 4.63 (br s,
1H, NH), 4.82-4.91 (m, 1H, CH), 4.95-5.04 (m, 1H, NH), 5.16 (m, 1H, NH), 7.07 (s, 1H,
ArH), 7.22-7.43 (m, 16H, ArH), 7.87 (s, 1H, ArH), 8.11 (dd, 1H, J = 8.1,1.5 Hz, ArH), 8.48
(dd, 1H, J = 4.8, 1.2 Hz, ArH), 8.80 (s, 1H, ArH).
m/z(ES)317(M++H).
Example 18
6-(5-Methyl-1H-benzimidazol-1-yl)-N-[(1S)-1-phenylethyl]pyrazin-2-amine and
6-(6-methyl-1H-benzimidazol-1-yl)-N-[(1S)-1-pheny]ethyl]pyrazin-2-amine

In a procedure analogous to example 12, reaction of
6-chloro-N-[(1S)-1-phenylethyl]pyrazin-2-amine (91mg, 0.39mmol) and
5-methylbenzimidazole (57mg, 0.43mmol) furnished the product (61mg, 148%) as a 1:1
mixture.
1H-n.m.r. (as 1:1 mixture) (CDCl3) d1.65 (d, 3H, CH3), 1.66 (d, 3H, CH3), 2.49 (s, 3H,
CH3), 2.50 (s, 3H, CH3), 5.01 (m, 1H, CH), 5.04 (m, 1H, CH), 5.56 (d, 1H, NH), 5.62 (d, 1H,
NH), 7.27-7.45 (m, 10H, Ph-H), 7.56-7.85 (m, 4H, benzimid-H), 7.94 (s, 1H, pyraz-H), 7.97
(s, 1H, pyraz-H), 8.02 (s, 2H, benzimid-H), 8.16 (s, 1H, pyraz-H), 8.17 (s, 1H, pyraz-H),
8.59 (s, 1H, benzimid-H), 8.70 (s, 1H, benzimid-H).
m/z(ES)330(M++H).
1Example 19
6-(2-Methyl-1H-benzimidazol-1-yl)-N-[(1S)-1-phenylethyl]pyrazin-2-amine

In a procedure analogous to example 12, reaction of
6-chloro-N-[(1S)-1-phenylethyl]pyrazin-2-amine (120mg, 0.51mmol) and
2-methyl-benzimidazole (75mg, 0.57mmol) furnished the product (18mg, 11%).
1H-n-m-r. (CDCl3) d1.63 (d, 3H, J = 6.9 Hz, CH3), 2.59 (s, 3H, CH3), 4.93 (m, 1H,
CH), 5.89 (br s, 1H, NH), 7.16-7.49 (m, 7H, ArH), 7.93 (d, 2H, J = 8.1 Hz, Ar-H), 8.00 (br s,
1H, pyraz-H), 8.23 (br s, 1H, pyraz-H).
m/z(ES)330(M++H).
Example 20
N-(2-Hydroxyethyl)-1H-benzimidazole-5-carboxamide

To a stirred suspension of benzimidazole-5-carboxylic acid (0.40g, 2.50mmol) in
benzene (8mL) was added thionyl chloride (2mL) dropwise at room temperature. The
whole mixture was then heated under reflux for 5h. Benzene and thionyl chloride was
evaporated off under reduced pressure, and the obtained acid chloride was suspended in
tetrohydrofuran. To this was the added 2-hydroxyethyl amine dropwise at ODC, and the
resultant mixture was then stirred at room temperature overnight. The solvent was then
decanted, and the residue was washed with diethyl ether (40mL). The residue oil was
then treated with cold water (5mL), and the aqueous solution was carefully decanted.
This step was repeated, and now the amide was obtained as a pale brown solid, 0.19g
(37%).
1H-n.m.r. (CDCl3) d 3.30-3.37 (m, 2H, CH2), 3.51-3.55 (m, 2H, CH2), 4.69 (s, 1H,
OH), 7.59 (d, 1H, J = 8.1Hz, ArH), 7.73 (d, 1H, J = 8.4Hz, ArH), 8.13 (br s, 1H, NH), 8.30 (s,
1H, H-4), 8.35 (br s, 1H, H-2), 12.47 (br s, 1H, CONH).
Example 21
1-[6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl]-N-(2-hydroxyethyl)-1H-benzimidazole-
5-carboxamide

In a procedure analogous to example 12, reaction of
6-chloro-N-[(1S)-1-phenylethyl]pyrazin-2-amine (91mg, 0.39mmol) and benzimidazole 5-
carboxylic acid (2-hydroxyethyl)amide (88mg, 0.43mmol) furnished the product which
was purified by column chromatography (22mg, 14%).
1H-n.m.r. (CDCl3) d1.63 (d, 3H, J = 6.9 Hz, CH3), 3.68 (2H, dt, J = 5.1 Hz, CH2NH),
3.91 (2H, t, J = 5.1 Hz, CH2OH), 4.98 (m, 1H, CH), 5.50 (d, 1H, J = 6.0 Hz, NH), 7.15 (1H, t, J
= 5.1 Hz, CONH), 7.28-7.41 (m, 5H, Ar-H), 7.60 (d, 1H, J = 8.4 Hz, benzimid-H ), 7.73 (d,
1H, J = 8.4 Hz, benzimid-H), 7.88 (s, 1H, pyraz-H), 8.05 (s, 1H, pyraz-H), 8.20 (s, 1H,
benzimid-H), 8.40 (s, 1H, benzimid-H). m/z (ES) 403 (M++H).
Example 22
[1-(6-{[(1S)-1-Phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-5-yl]methanol and
[1-(6-{[(R)-1-phenylethyI]amino}pyrazin-2-yl)-1H-benzimidazol-6-yl]methanol

In a procedure analogous to example 12, reaction of
6-chloro-N-[(15)-1-phenylethyl]pyrazin-2-amine (1.07g, 4.6mmol) and 5-hydroxymethyl
benzimidazole (0.68 g, Y 4.60mmol) furnished the two products separated by column
chromatography: 5-hydroxymethyl isomer (34mg, 2%); 6-hydroxymethyl isomer (36mg,
2%).
(5-hydroxymethyl isomer) 1H-n.m.r. (CDCl3) d1.65 (d, 3H,J = 6.9 Hz, CH3), 4.82 (s,
2H, CH2OH), 5.03 (m, 1H, CH), 5.26 (d, 1H, J = 6.0 Hz, NH), 7.25-7.40 (m, 6H, Ar-H), 7.68
(d, 1H, J = 8.4 Hz, benzimid-H), 7.81 (s, 1H, benzimid-H), 7.85 (s, 1H, pyraz-H), 8.12 (s,
1H, pyraz-H), 8.39 (s, 1H, benzimid-H). m/z (ES) 346 (M++H).
(6-hydroxymethyl isomer) 1H-n.m.r. (CDCl3) d1.63 (d, 3H, J = 6.9 Hz, CH3), 4.79 (s,
2H, CH2OH), 5.03 (m, 1H, CH), 5.42 (d, 1H, J = 6.3 Hz, NH), 7.25-7.42 (m, 6H, Ar-H), 7.78
(d, lH,J = 8.4 Hz, benzimid-H), 7.81 (s, 1H, pyraz-H), 7.90 (s, 1H, benzimid-H), 8.10 (s,
1H, pyraz-H), 8.35 (s, 1H, benzimid-H). m/z (ES) 346 (M++H).
Example 23
N-Methyl-1H-benzimidazole-5-carboxamide

In a procedure analogous to example 20, reaction of benzimidazole-5-carboxylic acid and
aqueous methylamine furnished the product in 67% yield.
1H-n.m.r. (CDCl3) d 2.79 (s, 3H, CH3), 7.59 (d,1H,J = 8.4Hz, H-6), 7.71 (dd, 1H, J =
8.4, 1.2Hz, H-7), 8.10 (s, 1H, H-4), 8.29 (s, 1H, H-2), 8.36 (br s, 1H, NH), 12.56 (br s, 1H,
CONH).
Example 24
N-Methyl-1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazole-
5-carboxamide and N-Methyl-1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-
1H-benzimidazole-6-carboxamide

In a procedure analogous to example 12, reaction of
6-chloro-N-[(1S)-1-phenyIethyl)pyrazin-2-amine (91mg, 0.39mmol) and
5-carboxybenzimidazole N-methyl amide (75mg, 0.43mmol) furnished the two products
separated by column chromatography: 5-isomer (23mg,, 16%); 6-isomer (38mg, 26%).
(5-isomer) 1H-am.r. (CDCl3) d1.67 (d, 3H, J = 6.9 Hz, CH3), 3.06 (d, 3H, J = 4.5Hz,
NCH3), 5.01 (m, 1H, CH), 5.58 (br s, 1H, NH), 6.43 (br s, 1H, CONH), 7.30-7.43 (m, 5H,
Ph-H), 7.71 (d, 1H, J=8.4Hz, benzimid-H), 7.84-7.88 (m, 2H, benzimid-H, pyraz-H), 8.19 (s,
1H, pyraz-H), 8.53 (s, 1H, benzimid-H), 8.57 (s, 1H, benzimid-H).
m/z(ES)373(M++H).
(6-isomer) 1H-n.m.r. (CDCl3) d1.65 (d, 3H, J= 6.9 Hz, CH3), 3.07 (d, 3H, J = 4.5Hz,
NCH3), 5.02 (m, 1H, CH), 5.31 (d, 1H, J = 4.8Hz, NH), 6.30 (br s, 1H, CONH), 7.31-741 (m,
5H, Ph-H), 7.68 (d, 1H, J = 8.4Hz, benzimid-H), 7.81 (d, 1H, J=8.4Hz, benzimid-H), 7.92 (s,
1H, pyraz-H), 8.13 (s, 1H, pyraz-H), 8.18 (s, 1H, benzimid-H), 8.49 (s, 1H, benzimid-H).
m/z (ES) 373 (M++H).
Example 25
1-(6-{[(1S)-1-Phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-5-amine and
1-(6-([(1S)-1-phenylethyl]anuno}pyrazin-2-yl)-1H-benzinudazo]-6-amine

To a stirred solution of 5-amino-benzimidazole (290mg, 2.2mmol) in anhydrous
DMF (10mL) 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)-1-phenylethyl]pyrazin-2-amine (470mg) in DMF (5mL) and the resulting
mixture was then heated at reflux for 48h. The DMF was removed under reduced
pressure and the residue diluted with chloroform. The organic layer was washed with
aqueous Na2CO3, dried (Na2SO4) 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 unreacted starting material. The
higher Rf fraction was assigned as the 6-isomer (276mg, 42%). 1H-n.m.r. (CDCl3) d1.64 (d,
3H,J = 6.9Hz, CH3), 2.90 (br s, 2H, NH2), 5.05 (m, 1H, CH), 5.21 (d, 1H, NH), 6.70 (dd, 1H,
J = 8.7, 2.1Hz, benzimid-H), 6.97 (d, lH,J = 1.8Hz, benzimid-H), 7.28-7.43 (m, 5H, Ph-H),
7.58 (d, lH,J = 8.4Hz, benzimid-H), 7.84 (s, 1H, pyraz-H), 8.08 (s, 1H, pyraz-H), 8.21 (s,
1H, benzimid-H). m/z (ES) 331 (M++H). The lower fraction was assigned as the 5-isomer
(170mg, 26%), 1H-n.m.r. (CDCl3) d1.64 (d, 3H,J = 6.9Hz, CH3), 2.85 (br s, 2H, NH2), 5.01
(m, 1H, CH), 5.19 (d, 1H, NH), 6.70 (dd, 1H, J = 8.7, 2.1Hz, benzimid-H), 7.11 (d, 1H, J =
1.8Hz, benzimid-H), 7.29-7.40 (m, 5H, Ph-H), 7.51 (d, 1H,J=8.7Hz, benzimid-H), 7.81 (s,
1H, pyraz-H), 8.10 (s, 1H, pyraz-H), 8.32 (s, 1H, benzimid-H).
m/z (ES) 331 (M++H).
Example 26
N-Benzyl-1H-benzimidazole-5-carboxamide

In a procedure analogous to example 20, reaction of benzimidazole-5-carboxylic
acid (400mg, 2.50mmol) and benzylamine furnished the product (410mg, 66%).
1H-n.m.r. (CDCl3) d 4.56 (d, 2H, J = 5.8Hz, CH2), 7.13-7.31 (m, 5H, Ph-H), 7.59 (d,
1H, J = 8.5Hz, H-6), 7.79 (dd, 1H, J = 8.5Hz, H-7), 7.96 (br s, 1H, CONH), 8.21 (s, 1H, H-4),
8.29 (s,1H, H-2).
Example 27
N-Benzyl-1-[6-([(1S)-1-phenylethyl]amino)pyrazin-2-yl]-lH-benzimidazole-5-carboxamid
e and N-Benzyl-1-[6-([(TS)-1-phenylethyl]amino)pyrazin-2-yl-lH-benzimidazole-
6-carboxamide

In a procedure analogous to example 25, reaction of 6-chloro-N-
[(1S)-1-phenylethyl]pyrazin-2-amine (91mg, 0.39mmol) and
N-benzyl-1H-benzimidazole-5-carboxamide (108mg, 0.43mmol) furnished the product
(132mg, 75%) as a 3:1 mixture of regioisomers.
1H-n.m.r. (as 3:1 mixture) (CDCl3) d1.62 (d, 3H, J = 6.9Hz, CH3), 1.63 (d, 3H, J =
6.9Hz, CH3), 4.68 (d, 2H, J = 5.4 Hz, CH2), 4.70 (d, 2H, J = 5.4 Hz, CH2), 4.95-5.04 (m, 1H,
CH), 5.55 (d, 1H, NH), 5.61 (d, 1H, NH), 6.80 (t-like, 1H, CONH), 6.92 (Mike, 1H, CONH),
7.22-7.40 (m, Ph-H, Ph-H), 7.66 (d, 1H, J = 8.4Hz, benzimid-H), 7.71 (d, 1H, J = 8.4Hz,
benzimid-H), 7.80-7.85 (m, benzimid-H), 7.91 (s, 1H, benzimid-H), 8.05 (s, 1H, pyraz-H),
8.13 (s, 1H, pyraz-H), 8.25 (s, 1H,, benzimid-H), 8.38 (s, 1H, pyraz-H), 8.46 (s, 1H,
benzimid-H),8.59 (s, 1H, pyraz-H).
m/z(ES)449(M++H).
Example 28
N-Phenyl-1H-benzimidazole-5-carboxamide

In a procedure analogous to example 20, reaction of benzimidazole-5-carboxylic acid
(400mg, 5.50mmol) and aniline (510mg, 5.50mmol) furnished the product (370mg, 63%).
1H-n.m.r. (CDCl3) d 6.83 (m, 1H, Ar-H), 7.08 (m, 2H, Ar-H), 7.53-7.74 (m, 4H,
Ar-H), 8.13-8.31 (m, 2H, Ar-H + CONH), 9.57 (s, 1H, H-2).
Example 29
1-(6-{[(1S)-1-Phenylethyl]amino}pyrazin-2-yl)-N-phenyl-1H-benzimidazole-5-
carboxamide and 1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-N-phenyl-1H-
benzimidazole-6-carboxamide

In a procedure analogous to example 25, reaction of 6-chloro-N-
[(1S)-1-phenylethyl]pyrazin-2-amine (91mg, 0.39mmol) and N-phenyl-
1H-benzimidazole-5-carboxamide (102mg, 0.43mmol) furnished the product (100mg,
59%) as a 2:1 mixture of regioisomers.
1H-n.m.r. (as 2:1 mixture) (CDCl3) 81.61 (d, 3H, J = 6.9Hz, CH3), 1.62 (d, 3H, J =
6.9Hz, CH3), 4.94-5.03 (m, CH, CH), 5.55 (d, 1H, NH), 5.62 (d, 1H, NH), 7.18-7.45 (m, Ph-H,
Ph-H), 7.61-7.83 (m, benzimid-H and Ph-H)), 7.85 (s, 1H, pyraz-H), 7.90 (s, 1H, pyraz-H),
8.06 (s, 1H, pyraz-H), 8.12 (s, 1H, pyraz-H), 8.33 (s, 1H, benzimid-H), 8.36 (s, 1H,
benzimid-H), 8.46 (s, 1H, benzimid-H), 8.49 (s, 1H, CONH), 8.52 (s, 1H, CONH), 8.57 (s,
1H, benzimid-H).
m/z(ES)435(M++H).
Example 30
5-[(4-Methylpiperazin-1-yl)carbonyl]-1H-benzimidazole

In a procedure analogous to example 20, reaction of benzimidazole-5-carboxylic
acid (400mg, 2.50mmol) and N-methylpiperazine (550mg, 5.0mmol) furnished the
product (380mg, 63%).
1H-n.m.r. (CDCl3) d2.33 (s, 3H, CH3), 2.44(br s, 4H, CH2), 3.70 (br s, 4H, CH2), 7.25
(s, 1H, ArH), 7.66 (br s, 1H, ArH), 7.87 (s, 1H, ArH), 11.88 (s, 1H, H-2).
m/z (ES) 245 (M++H).
Example 31
6-{5-[(4-Methylpiperazin-1-y])carbonyl]-1H-benzimidazol-1-yl}-N-[(1S)-1-
phenylethyl]pyrazin-2-amine and 6-{6-[(4-Methylpiperazin-1-y])carbonyl]-
1H-benzimidazol-1-yl}-N-[(1S)-1-phenylethyl]pyrazin-2-amine

In a procedure analogous to example 25, reaction of
6-chloro-N-[(1S)-1-phenylethyl]pyrazin-2-amine (91mg, 0.39mmol) and
5-[(4-methylpiperazin-1-yl)carbonyl]-1H-benzimidazole (105mg, 0.43mmol) furnished the
product as a mixture of regioisomers which were separated by column chromatography:
6-isomer (58mg, 34%); 5-isomer (68mg, 40%).
(5 isomer) 1H-n.m.r. (CDCl3) d1.63 (d, 3H, J = 6.9Hz, CH3), 2.33 (s, 3H, NCH3), 2.44
(br s, 4H, CH2), 3.67 (br s, 4H, CH2), 5.01 (m, 1H, CH), 5.48 (d, 1H, J = 6.0 Hz, NH),
7.26-7.38 (m, 6H, Ar-H), 7.65 (d, 1H,J = 8.4Hz, benzimid-H), 7.85 (s, 1H, benzimid-H), 7.89
(s, 1H, pyraz-H), 8.09 (s, 1H, pyraz-H), 8.41 (s, 1H, benzimid-H).
m/z(ES)442(M++H).
(6 isomer) 1H-n.m.r. (CDCl3) d 1.63 (d, 3H,J = 6.9Hz, CH3), 2.31 (s, 3H, NCH3), 2.43
(br s, 4H, CH2), 3.4-3.9 (br m, 4H, CH2), 4.99 (m, 1H, CH), 5.54 (d, 1H, J = 6.0 Hz, NH),
7.23-7.39 (m, 6H, Ar-H), 7.83-7.85 (m, 2H, Ar-H, pyraz-H), 8.07 (s, 1H, benzimid-H), 8.12
(s, 1H, pyraz-H), 8.44 (s, 1H, benzimid-H). m/z (ES) 442 (M++H).
Example 32
N-[1-(6-{[(1S)-1-Phenylethyl]amino}pyrazin-2-yl)-1H-benzinidazol-5-yl]acetamide

To a stirred solution of 2-(S-a-methylbenzylamino)-6-(5-amino-
benzimidazo-1-yl)-pyrazine (66mg, 0.2mmol) in anhydrous THF (2mL) under N2 was
added triethylamine (41mg, 0.4mmol). The solution was cooled at 0°C and to this was
added acetyl chloride (17mg, 0.22mmol) and the resulting mixture then stirred at RT.
After 18h the solution was poured into water (30mL) and the product extracted into
chloroform (2 x 20mL). The combined organic layers were dried (Na2SO4) and the solvent
removed under reduced pressure to furnish the crude product as a pale yellow solid.
Column chromatography using dichloromethane-methanol (200:15) as eluant separated
the product as a pale yellow solid (38mg).
1H-n.m.r. (CDCL3) d1.63 (d, 3H, J=6.6Hz, CH3), 2.21 (s, 3H, CH3), 5.00 (m, 1H, CH),
5.43 (d, 1H, J = 5.7Hz, NH), 7.27-7.38 (m, 5H, ArH), 7.49 (d, 1H, J = 9.0Hz, benzimid-H),
7.61 (d, 1H, J = 9.0Hz, benzimid-H), 7.74 (br s, 1H, CONH), 7.84 (s, 1H, pyraz-H), 7.90 (s,
1H, benzimid-H), 8.11 (s, 1H, pyraz-H), 8.36 (s, 1H, benzimid-H).
m/z(ES)373(M++H).
Example 33
N-[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-6-yl]acetamide

In a procedure analogous to example 32, reaction of 2-(S-a-methylbenzylamino)-6-
(6-amino-benzimidazo-1-yl)-pyrazine (66mg, 0.20mmol) and acetyl chloride (17mg,
0.22mmol) furnished the product (70mg, 94%) after chromatography.
1H-n.m.r. (CDCl3) d1.65 (d, 3H,J = 6.9Hz, CH3), 2.22 (s, 3H, CH3), 5.07 (m, 1H, CH),
5.29 (d, lH,J = 6.3Hz, NH), 7.28-7.43 (m, 6H, ArH, benzimid-H), 7.72 (d, 1H, J = 8.7Hz,
benzimid-H), 7.84 (s, 1H, pyraz-H), 8.12 (s, 1H, pyraz-H), 8.27 (s, 1H, benzimid-H), 8.34 (s,
1H, benzimid-H).
m/z(ES)373(M++H).
Example 34
N-[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-5-yl]benzamide

In a procedure analogous to example 32, reaction of
1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-5-amine(33mg,
0.10mmol) and benzoyl chloride (14mg, 0.10mmol) furnished the product (23mg, 53%)
after chromatography.
1H-n.m.r. (CDCl3) d1.64 (d, 3H, J = 6.8Hz, CH3), 5.02 (m, 1H, CH), 5.33 (d, 1H, J =
4.5Hz, NH), 7.27-7.68 (m, 10H, ArH), 7.85 (s, 1H, pyraz-H), 7.90-7.93 (m, 2H, benzimid-H),
8.03 (s, 1H, benzimid-H), 8.06 (s, 1H, CONH), 8.10 (s, 1H, pyraz-H), 8.38 (s, 1H,
benzimid-H).
m/z(ES)435(M++H).
Example 35
N-[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimadazol-6-yl]benzamide

In a procedure analogous to example 32, reaction of
1-(6-{[(1S)-1-phenylethyl]amino)pyrazin-2-yl)-1H-benzirnidazol-6-amine(50mg,
0.15mmol) and benzoyl chloride (21mg, 0.15mmol) furnished the product (50mg, 76%)
after chromatography.
1H-n.m.r. (CDCl3) d1.64 (d, 3H, J = 6.6Hz, CH3), 5.11 (m, 1H, CH), 5.31 (d, 1H, J =
6.6Hz, NH), 7.16-7.58 (m, 10H, ArH), 7.78 (d, 1H,J = 8.4 Hz, benzimid-H), 7.83 (s, 1H,
benzimid-H), 7.90 (d, 1H, J =8.1 Hz, benzimid-H), 7.97 (s 1H, CONH), 8.15 (s, 1H,
pyraz-H), 8.35 (s, 1H, pyraz-H), 8.51 (s, 1H, benzimid-H).
m/z(ES)435(M++H).
Example 36
N-[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-5-yl]isonicotinamide

In a procedure analogous to example 32, reaction of
1-(6-{[(1S)-1-phenylethyl]amino)pyrazin-2-yl)-1H-benzimidazol-5-amine(33mg,
0.10mmol) and isonicotinoyl chloride hydrochloride (20mg, 0.11mmol) furnished the
product (10mg, 23%) after chromatography.
1H-n.m.r. (CDCl3) 51.56 (d, 3H, J = 6.9Hz, CH3), 4.98 (m, 1H, CH), 6.21 (d, 1H, J =
6.0Hz, NH), 7.11-7.37 (m, 5H, ArH), 7.59 (d, 1H, J = 9.0Hz, benzimid-H), 7.65 (d, 1H, J =
8.7Hz, benzimid-H), 7.81 (d, 2H,J = 5.7Hz, pyrid-H), 7.87 (s, 1H, pyraz-H), 8.01 (s, 1H,
pyraz-H), 8.10 (s, 1H, benzimid-H), 8.32 (s, 1H, benzimid-H), 8.72 (d, 2H,J=5.7Hz,
pyrid-H), 9.58 (s, 1H, CONH).
m/z(ES)436(M++H).
Example 37
N-[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-6-yl]isonicotinamide

In a procedure analogous to example 32, reaction of
1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-6-amine(33mg,
O.lOmmol) and isonicotinoyl chloride hydrochloride (20mg, 0.11mmol) furnished the
product (10mg, 23%) after chromatography.
1H-n.m.r. (CDCl3) d1.63 (d, 3H,J = 6.9Hz, CH3), 5.11 (m, 1H, CH), 5.40 (d, 1H,J =
6.3Hz, NH), 7.18-7.42 (m, 6H, ArH, CONH), 7.73-7.79 (m, 3H, benzimid-H), 7.83 (s, 1H,
pyraz-H), 8.10 (s, 1H, pyraz-H), 8.36 (br s, 2H, pyrid-H), 8.48 (s, 1H, benzimid-H), 8.79 (d,
2H,J =5.7Hz, pyrid-H).
m/z(ES)436(M++H).
Example 38
N-[1-(6-{[(R)-1-phenylethyl]anuno}pyrazin-2-yl)-1H-benzimidazol-5-yl]nicotinamide

In a procedure analogous to example 32, reaction of
1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-5-amine(33mg,
0.10mmol) and nicotinoyl chloride hydrochloride (20mg, 0.11mmol) furnished the
product (17mg, 39%) after chromatography.
1H-n.m.r. (CDCl3) d1.48 (d, 3H, J = 6.9Hz, CH3), 4.91 (m, 1H, CH), 6.59 (d, 1H, J =
5.7 Hz, NH), 7.11-7.32 (m, 6H, ArH, pyrid-H), 7.58 (d, 1H, J = 9.0 Hz, benzimid-H), 7.83 (s,
1H, benzimid-H), 7.91 (s, 1H, pyraz-H), 8.07 (s, 1H, benzimid-H), 8.20(d, 1H,J = 8.1 Hz,
pyrid-H), 8.24 (s, 1H, pyraz-H), 8.61 (d, 1H,J = 4.6 Hz, pyrid-H), 9.12 (s, 1H, pyrid-H), 9.80
(s, 1H, CONH).
m/z(ES)436(M++H).
Example 39
M-[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-6-yl]nicotinamide

In a procedure analogous to example 32, reaction of
1-(6-{[(15)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-6-amine(33mg,
0.10mmol) and isonicotinoyl chloride hydrochloride (20mg, 0.11mmol) furnished the
product (27mg, 62%) after chromatography.
1H-n.m.r. (CDCl3)d1.62 (d, 3H, J = 6.6 Hz, CH3), 5.12 (m, 1H, CH), 5.41 (d, 1H, J =
6.0 Hz, NH), 7.17-7.21 (m, 1H, Ar-H), 7.26-7.29 (m, 1H, Ar-H), 7.39-7.45 (m, 4H, ArH,
pyrid-H), 7.76 (d, 1H,J = 8.7 Hz, benzimid-H), 7.82 (s, 1H, pyraz-H), 8.11 (s, 1H, pyraz-H),
8.21 (d, 1H, J = 8.1 Hz, pyrid-H), 8.33 (s, 1H, CONH). 8.35 (s, 1H, Ar-H), 8.47 (3, 1H,
pyrid-H), 8.76 (1H, d, J = 4.5 Hz, pyrid-H), 9.13 (s, 1H, pyrid.-H).
m/z(ES)436(M++H).
Example 40
N-[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazo]-5-yl]
pyrazine-2-carboxamide

In a procedure analogous to example 32, reaction of
1-(6-([(1S)-1-phenylethyl]arnino}pyrazin-2-yl)-1H-benzirnidazol-5-amine(33mg,
0.10mmol) and pyrazine-2-carbonyl chloride (0.11mmol) (prepared by reacting
pyrazine-2-carboxylic acid with oxalyl chloride in didoromethane at room temperature)
furnished the product (24mg, 55%) after chromatography.
1H-n.m.r. (CDCl3) d1.65 (d, 3H, J = 6.9 Hz, CH3), 5.04 (m, 1H, CH), 5.35 (d, 1H, J =
6.0 Hz, NH), 7.28-7.41 (m, 5H, Ar-H, pyrid-H), 7.71 (s, 1H, pyraz-H), 7.87 (s, 1H, pyraz-H),
8.12 (s, 1H, pyraz-H), 8.21 (s, 1H, benzimid-H), 8.40 (s, 1H, pyraz-H), 8.61-8.62 (m, 1H,
benzimid-H), 8.81 (d, 1H, J = 2.4 Hz, benzimid-H), 9.55 (d, 1H, J = 1.2 Hz, benzimid-H),
9.78 (s, 1H, CONH).
m/z(ES)437(M++H).
Example 41
N-[1-(6-([(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-6-yl]
pyrazine-2-carboxamide

In a procedure analogous to example 32, reaction of
1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-6-amine(33mg,
0.10mmol) and isonicotinoyl chloride hydrochloride (0.11mmol) (prepared by reacting
pyrazine-2-carboxylic acid with oxalyl chloride in didoromethane at room temperature)
furnished the product (28mg, 64%) after chromatography.
1H-n.m.r. (CDCl3) d1.67 (d, 3H, J = 6.9 Hz, CH3), 5.18 (m, 1H, CH), 5.39 (d, 1H, J =
6.3 Hz, NH), 7.23-7.46 (m, 6H, Ar-H, pyrid-H), 7.82 (d, 1H, J = 9.0 Hz, benzimid-H), 7.84
(s, 1H, pyraz-H), 8.18 (s, 1H, pyraz-H), 8.39 (s, 1H, pyraz-H), 8.61-8.62 (m, 1H, pyraz-H),
8.81-8.84 (m, 2H, Ar-H), 9.52 (d, 1H, J = 1.2 Hz, benzimid-H), 9.81 (s, 1H, CONH).
m/z(ES)437(M++H).
Example 42
2-Methoxy-N-[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-bendmidazol-5-yl]
acetamide

In a procedure analogous to example 32, reaction of
1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-5-amine(33mg,
0.10mmol) and methoxyacetyl chloride (12mg, 0.11mmol) furnished the product (20mg,
50%) after chromatography.
1H-n.m.r. (CDCl3) d1.64 (d, 3H, J =6.9Hz, CH3), 3,55 (s, 3H, OCH3), 4.07 (s, 2H, CH2),
5.02 (m, 1H, CH), 5.27 (d, 1H, J = 6.3 Hz, NH), 7.30-7.40 (m, 5H, Ar-H), 7.54 (dd, 1H, J =
8.7,1.5 Hz, benzimid-H), 7.66 (d, 1H, J = 8.7 Hz, benzimid-H), 7.85 (s, 1H, pyraz-H), 8.01
(s, 1H, benzimid-H), 8.11 (s, 1H, pyraz-H), 8.35 (s, 1H, CONH), 8.38 (s, 1H, benzimid-H).
m/z(ES)403(M++H).
Example 43
2-Methoxy-N-[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-6-yl]
acetamide

In a procedure analogous to example 32, reaction of
1-(6-{[(1S)-1-phenylethyI]arnino}pyrazin-2-yl)-1H-benzimidazol-6-amine(33mg/
O.lOmmol) and methoxyacetyl chloride (20mg, 0.11mmol) furnished the product (10mg,
25%) after chromatography.
1H-n.m.r. (CDCl3) d1.64 (d, 3H, J =6.9Hz, CH3), 3.54 (s, 3H, OCH3), 4.06 (s, 2H, CH2),
5.08 (m, 1H, CH), 5.36 (d, 1H, J = 6.3 Hz, NH), 7.24-7.42 (m, 6H, Ar-H), 7.76 (d, 1H, J = 8.7
Hz, benzimid-H), 7.81 (s, 1H, pyraz-H), 8.14 (s, 1H, pyraz-H), 8.35 (s, 1H, benzimid-H),
8.39 (s, 1H, CONH), 8.52 (d, 1H,J = 1.5 Hz, benzimid-H).
m/z(ES)403(M++H).
Example 44
N-[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-6-yl]-
2,2-dimethylpropanamide

To a stirred solution of 2-(benzylamino)-6-(5-amino-benzimidazo-1-yl)-pyrazine
(33mg, 0.1mmol) in anhydrous THF (2mL) under N2 was added triethylamine (381,
0.3mmol). The solution was cooled at 0°C and to this was added pivalic acid (12mg,
0.11mmol) and EDC (23mg, 0.12mmol) and the resulting mixture then stirred at RT. After
64h the solution was diluted with H2O and the mixture extracted with CHCl3 (2 x 15mL).
The combined organic layers were washed with 10% aqueous Na2CO3, dried (Na2SO4) and
the solvent removed in vacuo. The residue was purified by column chromatography
using dichloromethane-methanol (100:6) as eluant to separate the pur product (15mg).
1H-n.m.r. (CDCl3) d1.35 (s, 9H, 3CH3), 1.65 (d, 3H,J = 6.6Hz, CH3), 5.14 (m, 1H,
CH), 5.24 (d, 1H, J = 5.7Hz, NH), 7.13 (d, 1H, J = 8.7 Hz, Ar-H), 7.29-7.47 (m, 5H, ArH),
7.75 (d, 1H, J = 8.7 Hz, benzimid-H), 7.81 (s, 1H, pyraz-H), 8.17 (s, 1H, pyraz-H), 8.35 (s,
1H, benzimid-H), 8.69 (s, 1H, CONH).
m/z(ES)415(M++H).
Example 45
N-[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-y])-1H-benzimidazol-5-yl]-
2,2-dimethylpropanamide

In a procedure analogous to example 44, reaction of
1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-5-amine(33mg,
0.10mmol) and pivalic acid (12mg, 0.11mmol) furnished the product (12mg, 29%) after
chromatography.
1H-n.m.r. (CDCl3) d1.35 (s, 9H, CH3), 1.66 (d, 3H, J = 6.9 Hz, CH3), 5.14 (m, 1H, CH),
5.24 (d, 1H, J = 6.3 Hz, NH), 7.13 (d, 1H, J = 8.7 Hz, Ar-H), 7.29-7.47 (m, 6H, ArH), 7.75 (d,
1H,J = 8.7 Hz, benzimid-H), 7.81 (s, 1H, pyraz-H), 8.17 (s, 1H, pyraz-H), 8.35 (s, 1H,
benzimid.-H), 8.69 (s, 1H, CONH).
m/z(ES)415(M++H).
Example 46
6-{5-[(4-Methylpiperazin-1-yl)methyl]-1H-benzimidazol-1-yl}-N-[(1S)-1-
phenylethyl]pyrazin-2-amine

A solution of 6-{5-[(4-methylpiperazin-1-yl)carbonyl]-1H-
benzimidazol-1-yl}-N-[(1S)-1-phenylethyl]pyrazin-2-amine (22mg, 0.05mmol) in dry THF
(1mL) was added to a suspension of LiAlH4 (4mg, 0.1mmol) in THF (1mL) and the
mixture heated at reflux for 4h. Upon cooling to RT, the solution was treated
consecutively with H2O (lmL), aqueous NaOH (lmL, 2M) and H2O (5mL). The resulting
mixture was extracted with CHCl3 (2 x 10mL) and the combined organic layers dried
(Na2SO4). The solvent was removed under reduced pressure and the product purified by
flash chromatography using CH2Cl2-MeOH (10:1 ? 1:1) as eluant to afford the product as
a yellow solid (11mg, 52%).
1H-n.m.r. (CDCl3) d1.65 (d, 3H,J = 6.9Hz, CH3), 2.58 (s, 3H, NCH3), 2.81 (br s, 4H,
CH2), 2.90 (br s, 4H, CH2), 3.74 (s, 2H, NCH2), 5.03 (m, 1H, CH), 5.33 (d, 1H,J = 6.0 Hz,
NH), 7.25-7.42 (m, 6H, Ar-H), 7.67 (d, 1H, J = 8.4 Hz, benzimid-H), 7.77 (s, 1H,
benzimid-H), 7.87 (s, 1H, pyraz-H), 8.12 (s, 1H, pyraz-H), 8.39 (s, 1H, benzimid-H).
m/z(ES)428(M++H).
Example 47
1-[6-{[(1S)-1-Phenylethyl]amino}pyrazin-2-yl]-N-(pyridin-3-ylmethyl)-1H-benzimidazol-5
amine

In a procedure analogous to example 46, reaction of
N-[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-ben2imidazol-5-yl]nicotinamide
(33mg, 0.10mmol) with LiAlH4 (5.7mg, 0.15mmol) furnished the product (10mg, 24%)
after chromatography.
1H-n.m.r. (CDCl3) d1.64 (d, J = 6.9Hz, 3H, CH3), 4.42(s, 2H, CH2), 5.01(m, 1H, CH),
5.20(d, J = 6.0Hz, 1H, NH), 6.66 (dd, J = 8.7,2.1Hz, 1H, H-7"), 7.01 (d, J = 2.1Hz, 1H, H-4"),
7.25-7.45 (m, 6H, ArH), 7.51 (d, J = 8.7Hz, 1H, H-6"), 7.74 (d, J = 7.8Hz, pyridine-H), 7.80 (s,
1H, pyraz-H), 8.09 (s, 1H, pyraz-H), 8.30 (s,1H, H-2"), 8.53 (d, J = 3.6Hz, 1H, pyridine-H),
8.68(s, 1H, pyridine-H).
m/z(ES)422(M++H).
Example 48
N-[(1S)-1-(4-Bromophenyl)ethyl]-6-chloropyrazin-2-amine

In a procedure analogous to example 1, reaction of
4-bromo-a-S-methyl-benzylamine (877mg, 4.4mmol) and 2,6-dichloropyrazine (597mg,
4.0mmol) furnished the product (835mg, 67%).
1H-n.m.r. (CDCl3) d1.56 (d, J = 6.9Hz, 3H, CH3), 4.86 (m, 1H, CH), 5.0 (d, 1H, NH),
7.24 (AA"XX", 2H, Ar-H), 7.60 (s, 1H, pyraz-H), 7.81 (s, 1H, pyraz-H).
Example 49
6-Chloro-N-[(1S)-1-(4-methoxyphenyl)ethyl]pyrazin-2-amine

In a procedure analogous to example 1, reaction of
4-methoxy-a-S-methyl-benzylamine (700mg, 4.6mmol) and 2,6-dichloropyrazine (626mg,
4.2mmol) furnished the product (873mg, 79%).
1H-n.m.r. (CDCl3) d1.56 (d, J = 6.9Hz, 3H, CH3), 3.80 (s, 3H, OCH3), 4.84 (m, 1H,
CH), 5.01 (d, 1H, NH), 6.88 (AA"XX", 2H, Ar-H), 7.28 (AA"XX", 2H, Ar-H), 7.621 (s, 1H,
pyraz-H), 7.79 (s, 1H, pyraz-H).
Example 50
6-(1H-Benzimidazol-1-yl)-N-[(1S)-1-(4-bromophenyl)ethyl]pyrazin-2-amine

In a procedure analogous to example 25, reaction of
N-[(1S)-1-(4-bromophenyl)ethyl]-6-chloropyrazin-2-amine (125mg, 0.40mmol) and
benzimidazole (52mg, 0.44mmol) furnished the product (66mg, 42%).
1H-n.m.r. (CDCl3) d1.63 (d, 3H, J = 6.9 Hz, CH3), 4.99 (m, 1H, CH), 5.19 (d, 1H, J =
5.1 Hz, NH), 7.26-7.37 (m, 3H, Ar-H), 7.51 (AA"XX", 2H, Ar-H), 7.65 (d, 1H, J = 8.1 Hz,
benzimid-H), 7.83-7.86 (m, 2H, benzimid-H + pyraz-H), 8.17 (s, 1H, pyraz-H), 8.39 (s, 1H,
benzimid-H).
m/z(ES)396,394(M++H)
Example 51
6-(1H-Benzimidazol-1-yl)-N-[(1S)-1-(4-methoxyphenyl)ethyl]pyrazin-2-amine

In a procedure analogous to example 25, reaction of
6-chloro-N-[(1S)-1-(4-methoxyphenyl)ethyl]pyrazin-2-amine (105mg, 0.40mmol) and
benzimidazole (52mg, 0.44mmol) furnished the product (57mg, 41%).
1H-n.m.r. (CDCl3) d1.62 (d, 3H, J = 6.8 Hz, CH3), 3.80 (s, 3H, CH3), 4.99 (m, 1H,
CH), 5.31 (br d, 1H, J = 6.2 Hz, NH), 6.91 (AA"XX", 2H, Ar-H), 7.28-7.36 (m, 4H, Ar-H),
7.77 (d, 1H,J = 2.0 Hz, Ar-H), 7.78 (s, 2H, benzimid-H + pyraz-H), 8.13 (s, 1H, pyraz-H),
8.44 (s, 1H, benzimid-H).
m/z(ES)346(M++H)
Example 52
2-S-a-Methylbenzylamino)-6-(5-(N-methylpiperazin-4-yl-methyl)-benzimidazo-1-yl)-
pyrazine

A solution of
3-[6-(S-a-methylbenzylamino)-pyrazin-2-yl]-3H-benzoimidazole-5-carboxylic acid
N-methylpiperazinylamide (22mg, 0.05mmol) in dry THF (1mL) was added to a
suspension of LiAlH4 (4mg, 0.1mmol) in THF (1mL) and the mixture heated at reflux for
4h. Upon cooling to RT, the solution was treated consecutively with H2O (lmL), aqueous
NaOH (1mL, 2M) and H2O (5mL). The resulting mixture was extracted with CHCl3 (2 x
10mL) and the combined organic layers dried (Na2SO4). The solvent was removed under
reduced pressure and the product purified by flash chromatography using CH2Cl2-MeOH
(10:1 ? 1:1) as eluant to afford the product as a yellow solid (11mg, 52%).
1H-n.m.r. (CDCl3) d1.65 (d, 3H, J = 6.9Hz, CH3), 2.58 (s, 3H, NCH3), 2.81 (br s, 4H,
CH2), 2.90 (br s, 4H, CH2), 3.74 (s, 2H, NCH2), 5.03 (m, 1H, CH), 5.33 (d, 1H, J = 6.0 Hz,
NH), 7.25-7.42 (m, 6H, Ar-H), 7.67 (d, 1H, J = 8.4 Hz, benzimid-H), 7.77 (s, 1H,
benzimid-H), 7.87 (s, 1H, pyraz-H), 8.12 (s, 1H, pyraz-H), 8.39 (s, 1H, benzimid-H).
m/z(ES)428(M++H).
Example 53
1-{4-[2-(Diethylamino)ethoxy]phenyl}ethanone

A mixture of 4-bromoacetophenone (5g, 25 mmol), the diethylaminoethanol (3.5g,
38 mmol), K2CO3 (2g), copper powder (0.5g) and copper(I) iodide (2.5g) in DMSO (30 ml)
was heated at 120°C until TLC showed consumption of the starting material. After
cooling to RT, the mixture was poured onto aqueous NH3 (28 %, 100 ml), extracted with
CH2Cl2 (3 x 100 ml). The combined organic extracts were dried (Na2SO4), filtered and the
CH2Cl2 distilled in vacuo. The crude product was purified by column chromatography on
silica gel, eluting with EtOAc-MeOH (95:5) to furnish the pure compound (4.85g, 82%).
1H-n.m.r. (CDCl3) d0.98 (t, J = 7.2 Hz, 6H, NCH2CH3), 2.43 (s, 3 H, Ar-C-CH3),
2.57-2.50 (m, 4 H, NCH2CH3), 2.78 (t, J = 6.3 Hz, 2 H, ArOCH2CH2N), 4.00 (t,
J = 6.3 Hz, 2 H, ArO-CH2), 6.85 (d, J = 9 Hz, 2 H, ArH), 7.83 (d, J = 9 Hz, 2 H, ArH).
Example 54
1-[4-(4-Methylpiperazin-1-y])phenyl]ethanone

In a procedure identical to Example 53, 4-bromoacetophenone was reacted with
N-methylpiperazine in the presence of potassium carbonate, copper and copper iodide to
furnish the desired product in 82% yield.
1H-n.m.r. (CDCl3) d 2.35 (s, 3 H, Ar-C-CH3), 2.52 (s, 3 H, N-CH3), 2.55 (t, J = 5.1 Hz,
4 H, CH2-N-CH3), 3.37 (dd, J = 5.1, 5.1 Hz, 4H, ArNCH2), 6.89 (AA"XX", 2H, ArH), 7.89
(AA"XX", 2H, ArH).
Example 55
1-Pyridin-3-ylethanone oxime

To a solution of hydroxylamine hydrochloride (3.44g) in water (20 ml) was added
NaOH (20 %, 30 ml). The ketone (5g, 41 mmol) was added at once and the resulting
mixture was stirred at RT until TLC showed no ketone remained. The solvents were
distilled off in vacuo and the residue extracted with CH2C12 (3 x 100 ml) and dried
(Na2SO4). After filtration and removal of the solvent, the crude ketoxime was
recrystallised from CH2Cl2/n-hexane.
1H-n.m.r. (CDCl3) d2.31 (s, 3H, CH3), 7.33 (dd,J = 4.8,4.8 Hz, 1H, ArH), 7.97 (ddd, J
= 8.1,1.8,1.8Hz, 1H, ArH), 8.61 (dd, J = 5.1,1.8 Hz, 1H, ArH), 8.96 (d, J = 1.8 Hz, 1H,
ArH), 10.62 (s, 1 H, OH).
Example 56
1-(3-Chlorophenyl)ethanone oxime

A mixture of the ketone (2.0g, 13mmol), hydroxylamine hydrochloride (0.98g,
14mmol), NaOH (10%, 4ml), water (6.2ml) and EtOH (25 ml) was heated under reflux for
2 hours. Upon cooling in ice, the ketoxime precipitated and was collected by suction
filtration. The crude product was recrystallised from CH2Cl2/n-hexane (1.88g, 86%).
1H-n.m.r. (CDCl3) d 2.28 (s, 3H, CH3), 7.51 (s, 4H, ArH), 8.67 (s, 1H, OH).
Example 57
1-(3-Chlorophenyl)ethanamine

A mixture of the ketoxime (1g, 6mmol) and LiAlH4 (0.27g) in anhydrous THF (100
ml) was heated at reflux under dry N2 overnight. The reaction mixture was cooled in
ice-water and carefully quenched with H2O (60mL). The mixture was allowed to stir at
RT for half an hour, after which time it was filtered through Celite®. The inorganic salts
were washed with EtOAc (3 x 100 ml). The filtrate was concentrated under reduced
pressure, diluted with 2M HC1 (50ml) and the aqueous phase washed with Et2O (2 x
70ml). The aqueous phase was basified with 40% aqueous NaOH and the product
extracted with Et2O (3 x 50ml). The combined organic layers were washed with brine
(50ml) and dried (MgSO4). The solvents were removed in vacuo to afford the pure amine
(0.65g, 71%).
1H-n.m.r. (CDCl3) d1.38 (d,J = 6.6 Hz, 3H, CH-CH3), 1.63 (br s, 2 H, NH2), 4.13-4.06
(m, 1 H, CH-CH3), 7.23-7.18 (m, 3 H, ArH), 7.35 (s, 1 H, ArH).
Example 58
1-Pyridin-3-ylethanamine

To a mixture of the ketoxime (4.85g, 36 mmol) and Zn powder (12g) at 0°C was
slowly added, with vigorous stirring, concentrated HCl (50 ml). When the initial vigorous
reaction had subsided, the mixture was heated under reflux until TLC showed all the
ketoxime had been consumed. After cooling to RT, the strongly acidic mixture was
extracted with CH2Cl2 (2 x 75 ml). The reaction mixture was then made strongly basic
with 50% KOH solution. After removal of the solvent, the residue was extracted with
boiling MeOH (4 x 100 ml). The MeOH was distilled off to leave the crude amine which
was used in the ensuing reactions without further purification.
1H-n.m.r. (CDCl3) d1.07 (d, J = 6.6 Hz, 3 H, CH3), 1.37 (br s, 2H, NH2), 3.84 (q, J =
4.6Hz, 1H, CH-CH3), 6.93 (dd, J = 7.8,4.8 Hz, 1H, ArH), 7.38 (ddd, J = 7.8, 2.1,1.5 Hz, 1H,
ArH), 8.15 (dd, J = 4.8,1.5 Hz, 1H, ArH), 8.27 (d, J = 2.1 Hz, 1H, ArH).
Example 59
6-Chloro-N-[(1S)-1-(4-pyridin-3-ylphenyl)ethyl]pyrazin-2-amine

Under a nitrogen atmosphere a mixture of
N-[(15)-1-(4-bromophenyl)ethyl]-6-chloropyrazin-2-amine (0.117 g, 0.37 mmol),
pyridine-3-boronic acid 1,3-propanediol cyclic ester (67mg, 0.41 mmol),
tetrakis(triphenylphosphine)paIladium(0) (65 mg, 0.06 mmol) and toluene (4 mL) was
treated with 2M aqueous sodium carbonate solution (0.2 mL). The resulting mixture was
stirred vigorously whilst being heated under reflux for 24 hours. Upon cooling, the
solution was diluted with methanol and dichloromethane and the mixture dried (MgSO4)
and filtered. Removal of solvent in vacuo then yielded the crude product which was
purified by column chromatography using dichloromethane-diethyl ether (90:10) then
dichloromethane-methanol (99:1) as eluent (50 mg).
lH-n.m.r. (CDCl3) d1.61 (d, 3H,J = 6.9 Hz, CH3), 4.97 (m, 1H, CH), 5.42 (d, 1H,J =
6.3 Hz, NH), 7.33-7.37 (m, 1H, ArH), 7.42-7.56 (m, 4H, ArH), 7.66 (m, 1H, pyraz-H), 7.78 (s,
1H, pyraz-H), 7.83-7.86 (m, 1H, ArH), 8.58 (br s, 1H, pyrid-H), 8.83 (br s, 1H, pyrid-H).
m/z(ES)313,311(M++H).
Example 60
N-[1-(6-{[(1S)-1-Phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-5-yl]
methanesulfonamide

To a stirred solution of 2-(S-a-methylbenzylamino)-6-(5-amino-benzimidazo-1-yl)-
pyrazine (33mg, 0.1mmol) in anhydrous THF (2mL) under N2 was added triethylamine
(40mg, 0.4mmol). The solution was cooled at 0°C and to this was added
methanesulphonyl chloride (25mg, 0.2mmol) and the resulting mixture then stirred at RT.
After 16h the solution was poured into water (30mL) and the product extracted into
chloroform (2 x 15mL). The combined organic layers were was washed with 10% Na2CO3,
dried (Na2SO4) and the solvent removed under reduced pressure to furnish the crude
product as a pale yellow solid. Column chromatography, using
dichloromethane-methanol (100:6) as eluant, separated the product from the most polar
fractions as a pale yellow solid (16mg).
1H-n.m.r. (CDCl3) d1.65 (d, 3H, J = 6.9 Hz, CH3), 3.00 (s, 3H, CH3), 5.02 (m, 1H, CH),
5.27 (d, 1H, J = 6.0 Hz, NH), 7.21-7.40 (m, 6H, ArH), 7.64 (d, 1H, J = 8.7 Hz, benzimid-H),
7.69 (d, 1H, J = 1.9 Hz, benzimid-H), 7.88 (s, 1H, pyraz-H), 8.10 (s, 1H, pyraz-H), 8.41 (s,
1H, benzimid-H).
m/z(ES)409(M++H).
Example 61
N-[(1S)-1-Phenylethyl]-6-(5-pyridin-4-yl-1H-benzimidazol-1-yl)pyrazin-2-amine and
N-[(1S)-1-Phenylethyl]-6-(6-pyridin-4-yl-1H-benzimidazol-1-yl)pyrazin-2-amine

To a stirred solution of a 1:1 mixture of 6-(5-bromo-1H-benzimidazol-1-yl)-
N-[(1S)-1-phenylethyl]pyrazin-2-amine and 6-(6-bromo-1H-benzimidazol-1-yl)-
N-[(1S)-1-phenylethyl]pyrazin-2-amine (160mg, 0.4 mmol), pyridine-4-boronic acid
pinacol cyclic ester (91mg, 0.44 mmol) in toluene (5mL) was added
tetrakis(triphenyl-phosphine) palladium (46mg, 0.04mmol) followed by aqueous sodium
carbonate (0.22mL, 2M). The solution was then heated under reflux for 24h. After
standard work-up (see example 60) the products were obtained, as a 1:1 mixture by flash
chromatography using dichloromethane-methanol (100:6) as eluant.
1H-n.m.r. (as 1:1 mixture) (CDCl3) d1.66 (d,J = 6.9Hz, 3H, CH3), 5.07 (m, 1H, CH),
5.25 (d, J = 6.0Hz, 1H, NH), 7.29-7.42 (m, 5H, ArH), 7.52-7.64 (m, 3H), 7.78 (d, J = 8.7Hz),
7.88 and 7.90 (s, 1H, pyraz-H), 7.94 (d, 1H, J = 8.4Hz) 8.12 (s, 1H), 8.16 (s, 1H), 8.19 and
8.20 (s, 1H, pyraz-H), 8.44 and 8.45 (s,lH, H-2"), 8.68 (d,J = 3.6Hz, 1H, pyridine-H), 8.68
and 8.71 (d, lH,J = 8.7Hz, pyridine-H).
m/z(ES)393(M++H).
SCREENING
Compound Dilution
For screening purposes, compounds were diluted in 96 well plates at a
concentration of 20 µM. Plates were warmed at 37°C for 30 minutes before assay.
JAK Tyrosine Kinase Domain Production
JAK kinase domains were produced in the following manner:
JAK1
The kinase domain of human JAK1 was amplified from U937mRNA using the polymerase
chain reaction with the following primers:
XHOI-J1 5"-CCG CTC GAG ACT GAA GTG GAC CCC ACA CAT-3"
J1-KPNI 5"-CGG GGT ACC TTA TTT TAA AAG TGC TTC AAA-3"
JAK1 PCR products were cloned into the pFastBac HTb expression vector (Gibco) via the
Xho I and Kpn I sites. The JAK1 plasmid was then transformed into competent DHlOBac
cells (Gibco), and the recombinant baculovirus produced prepared for transfection into
Sf9 insect cells.
JAK2
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 GGA T-3"
jk2-NOTI 5"-ATA GTT TAG CGG CCG CTC AGA ATG AAG GTC ATT T-3"
JAK2 PCR products were cloned into the pFastBac HTc expression vector (Gibco) via the
Sal I and Not I sites. The JAK2 plasmid was then transformed into competent DH10Bac
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 AAA GGA CAG GGA GTG-3"
JAK3 PCR products were cloned into the pFastBac HTb expression vector (Gibco) via the
Xho I and Kpn I sites. The JAK3 plasmid was then transformed into competent DHlOBac
cells (Gibco), and the recombinant baculovirus produced prepared for transfection into
Sf9 insect cells.
TYK2
The kinase domain of humanTYK2 was amplified from A549 mRNA using the
polymerase chain reaction with the following primers:
HT2EK 5"-GGA GCA CTC GAG ATG GTA GCA CAC AAC CAG GTG-3"
ITY2.2R 5"-GGA GCA GGA ATT CCG GCG CTG CCG GTC AAA TCT GG-3"
TYK2 PCR products were cloned into pBlueBacHis2A (Invitrogen) via the EcoRI 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 (Invitrogen) 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 were 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, 10mM MgCl2, 150mM NaCl and 10µM-1mM ATP. The
biotinylated substrate biotin-EGPWLEEEEEAYGWMDF-NH2 (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 PY20. In the Alphascreen assay, Alphascreen
phosphotyrosine acceptor beads followed by streptavidin donor beads were added under
subdued light. The ELISA plates were read on a BMG Fluorostar, 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%.
Establishment of TEL:JAK cell lines
The coding region encompassing nucleotides 1-487 of TELwas amplified by PCR
using the oligonudeotides 5TEL (5" -GGA GGA TCC TGA TCT CTC TCG CTG TGA
GAC-3") and 3TEL (5"-AGGC GTC GAC TTC TTC TTC ATG GTT CTG-3") and U937
mRNA as template. A BamH I site was present into the 5TEL Primer, a Sal I site was
incorporated into the 3TEL primer. The regions encompassing the kinase domains of
JAK2 (nucleotides 2994-3914; ;JAK2F 5"-ACGC GTC GAC GGT GCC TTT GAA GAC
CGG GAT-3"; JAK2R 5"-ATA GTT TAG CGG CCG CTC AGA ATG AAG GTC ATT T-3"
) and JAK3 (nucleotides 2520-3469; JAK3F 5"-GAA GTC GAC TAT GCC TGC CAA GAC
CCC ACG ATC TT-3" ; JAK3R 5"-GGA TCT AGA CTA TGA AAA GGA CAG GGA GTG
GTG TTT -3") were generated by PCR using Taq DNA Polymerase ( Gibco/BRL) and
U937 mRNA as template. A Sail site was incorporated into the forward primer of JAK2
and JAK3, a Not I site was incorporated into the JAK2 reverse primer and a Xba I site was
added to the reverse primer of JAK3.
A TEL/Jak2 fusion was generated by digestion of the TELPCR product with
BamH I /Sal I, digestion of the JAK2 PCR product with Sal I/ Not I followed by ligation
and subcloning into the mammalian expression Vector pTRE 2 (Clontech) digested with
BamH I-Not I (pTELJAK2). For JAK3 Sal I/ Not I cleaved kinase domain PCR product
was ligated with BamH I /Sal I cleaved TELproduct followed by ligation into BamH
I/Not I cleaved pTRE2 (pTELJAK3).
The growth factor dependent myelomonocytic cell line BaF3 bearing the pTET-off
plasmid (Clontech) was transfected with either pTELJAK2 or pTELJAK3 and the cells
selected for factor independent growth. BaF 3 wild type cells were cultured in DMEM
10% FCS, 10% WEHI 3B conditioned medium. BaF3 TELJAK cells were cultured in
DMEM 10% Tet-System Approved FBS (without WEHI 3B conditioned medium).
Cellular assays were performed as follows:
Cell suspensions were prepared by harvesting cells from culture. (Cells used in
this test should be in later log phase growth and high viability.) Cells were diluted in
correct growth medium to 1.1x final concentration (from 50000 cell/mL to 200,000
cell/mL, depending on cell line).
Compounds to be tested were added (10µL, 10X final concentration) to a flat
bottom 96-well plate. The cellular suspension (90µL per well) was added, and the plate
incubated for 40 hr at 37 °C, 5% CO2. MTT (20 µL per well, 5mg/ mL in PBS) was added
and the plates were returned to the incubator for a further 6 hours. Lysis buffer (100 µL
per well, 10% SDS, 0.01N HC1) was added and the plate stored in the incubator overnight.
The plate was then read at 590 nm.
Results
The activity of a range of compounds is shown in Table 3. Compounds that
exhibited a capacity to inhibit 50% of JAK activity at a concentration of 50µM (measured
under standard conditions, see Methods), are designated as "+".
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
without 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.
WE CLAIM:-
1. Kinase inhibitor compounds of the general formula I:

or pharmaceutically acceptable salts, hydrates, solvates, crystal forms, diastereomers or
prodrugs thereof, wherein:
D is a heterocyclic ring selected from:

where X1, X2, X3, X4 are optionally substituted with a group R2, or one of X1, X2, X3,
X4 is N; R2 is 0-4 substituents independently chosen from H, halogen, C1-4 alkyl,
CH2F, CHF2, CF3, OCF3, aryl, hetaryl, C1-4alkylOC1-4 alkyl, C1-4alkylOaryl,
C1-4alkylNR3R4, CO2R3, CONR3R4, CONR3SO2R4, NR3R4, C1-4alkylNR3R4,
nitro, NR3COR4, NR5CONR3R4, NR3SO2R4, C1-4alkylNR3COR4,
C1-4alkylNR5CONR3R4, C1-4alkylNR3SO2R4; and R3, R4 are each independently
H, halogen, CH2F, CHF2, CF3, C1-4 alkyl, C1-4 alkyl cycloalkyl, C1-4 cyclohetalkyl,
aryl, C1-4 alkyl aryl, hetaryl, C1-4 alkyl hetaryl, or may be joined to form an optionally
substituted 3-8 membered (saturated or unsaturated) ring optionally containing an
atom selected from O, S, NR6; and R5 is selected from H, C1-4alkyl, halogen, CH2F,
CHF2, CF3, aryl or hetaryl; and R6 is selected from H, C1-4 alkyl, aryl, hetaryl, C1-4
alkyl aryl, C1-4 alkyl hetaryl.
R1 is H, C1-4 alkyl, C1-6 cycloalkyl;
Q is a bond, CH2, C1-4 alkyl, wherein when Q is a bond, W is not present;
A is aryl, hetaryl substituted with 0-3 substituents independently chosen from
halogen, C1-4 alkyl, CH2F, CHF2, CF3, OCF3, CN, NR8R9, aryl, hetaryl, C1-4alkylaryl,
C1-4alkylhetaryl, C1-4 alkylNR8R9, OC1-4alkylNR8R9, nitro, NR10CmNR8R9,
NR8COR9, NR10CONR8R9, NR8SO2R9, CO2R8 where R8 and R9 are each
independently H, C1-4 alkyl, aryl or which together form an optionally substituted 4-8
membered ring which may contain a heteroatom selected from O, S, NR11, where
Rl 1 is C1-4 alkyl, and R10 is selected from H, C1-4 alkyl;
W is selected from H, C1-4alkyl, C2-6alkenyl; where C1-4alkyl or C2-6alkenyl may be
optionally substituted with C1-4alkyl, OH, OC1-4alkyl, NR12R13; and R12, and R13
are each independently H, C1-4alkyl, 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, C1-4 alkyl.
A compound as claimed in claim 1 of the general formula II.

or pharmaceutically acceptable salts, hydrates, solvates, crystal forms, diastereomers
or prodrugs thereof, wherein:
D is a heterocyclic ring selected from:

where X1, X2, X3, X4 are optionally substituted with a group R2, or one of X1, X2, X3,
X4 is N; R2 is 0-4 substituents independently chosen from H, halogen, C1-4 alkyl,
CH2F, CHF2, CF3, OCF3, aryl, hetaryl, C1-4alkylOC1-4 alkyl, C1-4alkylOaryl,
C1-4alkylNR3R4, CO2R3, CONR3R4, CONR3SO2R4, nitro, NR3R4, C1-4
alkylNR3R4, NR3COR4, NR5CONR3R4, NR3SO2R4, C1-4alkylNR3COR4,
C1-4alkylNR5CONR3R4, C1-4alkylNR3SO2R4; and R3, R4 are each independently H,
halogen, CH2F, CHF2, CF3, C1-4 alkyl, C1-4 alkyl cycloalkyl, C1-4 cyclohetalkyl, aryl,
C1-4 alkyl aryl, hetaryl, C1-4 alkyl hetaryl, or may be joined to form an optionally
substituted 3-8 membered (saturated or unsaturated) ring optionally containing an
atom selected from O, S, NR6; and R5 is selected from H, C1-4 alkyl, halogen, CH2F,
CHF2, CF3, aryl or hetaryl; and R6 is selected from H, C1-4 alkyl, aryl, hetaryl, C1-4
alkyl aryl, C1-4 alkyl hetaryl;
R1 is H, C1-4 alkyl, C1-6 cycloalkyl;
W is H, C1-6 alkyl;
A is aryl, hetaryl substituted with 0-3 substituents independently chosen from
halogen, C1-4 alkyl, CH2F, CHF2, CF3, OCF3, CN, nitro, NR8R9, aryl, hetaryl,
C1-4alkylaryl, C1-4alkylhetaryl, C1-4alkylNR8R9, OC1-4 alkylNR8R9,
NR10C1-4NR8R9, NR8COR9, NR10CONR8R9, NR8SO2R9, CONR8R9, CO2R8
where R8 and R9 are each independently H, C1-4 alkyl, aryl or which together form
an optionally substituted 4-8 membered ring which may contain a heteroatom
selected from O, S, NR11, where Rl 1 is C1-4alkyl, and R10 is selected from H, C1-4
alkyl.
3. A compound as claimed in claim 1 or claim 2 where W is C1-4 alkyl wherein the
compound possesses S chirality at the chiral carbon bearing W.
4. A compound as claimed in claim 3 wherein the compound is a mixture of R and S
isomers and the mixture comprises at least 70% of the S isomer.
5. A compound as claimed in claim 4 wherein the compound comprises at least 80% of
the S isomer.
6. A compound as claimed in claim 4 wherein the compound comprises at least 90% of
the S isomer.
7. A compound as claimed in claim 4 wherein the compound comprises at least 95% of
the S isomer.
8. A compound as claimed in claim 4 wherein the compound comprises at least 99% of
the S isomer.
9. A compound wherein the compound is selected from the group consisting of 6-( 1H-
benzimidazol-1 -yl)-N- [(1R)-1 -phenylethyl]pyrazin-2-amine, N-benzyl-6-( 1H-
imidazol-1 -yl)pyrazin-2-amine, 6-( 1H-benzimidazol-1 -yl)-N- [(1S)-1 -(4-
methoxyphenyl)ethyl]pyrazin-2-amine, N-(4-Fluorobenzyl)-6-( 1H-imidazol-1-yl)
pyrazin-2-amine, 6-( 1H-benzimidazol-1-yl)-N- [(15)-1-(4-bromophenyl)ethyl]
pyrazin-2-amine, 6-( 1H-Imidazol-1-yl)-N-[(1R)-1-phenylethyl]pyrazin-2-amine, 1-
(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazole-6-carboxamide, 6-
(1H-Benzimidazol-1-yl)-N-benzylpyrazin-2-amine, 1 -(6-{[(1S)-1 -phenylethyl]
amino }pyrazin-2-yl)-1H-benzimidazole-5-carboxamide, 6-(1H-Benzimidazol-1-yl)-
N-(4-fluorobenzyl)pyrazin-2-amine, 6-{5-[(Morpholino-1-yl)carbonyl]-1H-
benzimidazol-1 -yl}-N-[(15)-1-phenylethyl]pyrazin-2-amine, 6-(1H-imidazo[4,5-b]
pyridin-1-yl)-N-[(1R)-1 -phenylethyl]pyrazin-2-aniine, 6- { 6- [(Morpholino-1 -yl)
carbonyl] -1 H-benzimidazol-1 -yl} -N-[(15)-1 -phenylethyl]pyrazin-2-amine, 6-( 1H-
imidazol-1 -yl)-N-(4-morpholin-4-ylphenyl)pyrazin-2-amine, N- [1-(6-{[(1S)-1-
Phenylethyl] amino } pyrazin-2-yl)-1H-benzimidazol-6-yl] cyclopropanecarboxamide,
N-[l-(6-{[(1S)-l -phenylethyl] amino} pyrazin-2-yl)-1H-benzimidazol-5 -yl]
nicotinamide, N-[1-(6-{[(1S)-1-Phenylethyl] amino }pyrazin-2-yl)-1H-benzimidazol-
5-yl]cyclopropanecarboxamide, 6-(1H-Benzimidazol-1-yl)-N-[(1R)-1-phenylethyl]
pyrazin-2-amine, 6-[6-(4,5-dihydro-1,3-oxazol-2-yl)-1H-benzimidazol-1-yl]-N-
[(1S)-1-phenylethyl]pyrazin-2-amine,N-[(1R)-1-Phenylethyl]-6-(4-phenyl-1H-
imidazol-1-yl)pyrazin-2-amine, 1-[6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl] -N-
(2-hydroxyethyl)-1H-benzimidazole-6-carboxamide, N-[1-(6-{[(1S)-1-phenylethyl]
amino } pyrazin-2-yl)-1H-benzimidazol-6-yl] methanesulfonamide, N-[1-(6-{[(1S)-1-
phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-5-yl]methanesulfonamide, N-[l-
(6- {[(1S)-1 -phenylethyl] amino} pyrazin-2-yl)-1H-benzimidazol-5 -yl]
isonicotinamide, 6-(1H-Imidazol-1-yl)-N-[(1S)-1-phenylethyl]pyrazin-2-amine, N-
[l-(6-{[(1S)-1-phenylethyl] amino}pyrazin-2-yl)-1H-benzimidazol-6-yl]
isonicotinamide, 6-(1H-Benzimidazol-1-yl)-N-[(15)-1-phenylethyl]pyrazin-2-amine,
6-[5-(4,5-dihydro-1,3-oxazol-2-yl)-1H-benzimidazol-1-yl]-N-[(1S)-1-phenylethyl]
pyrazin-2-amine, 1-[6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl]-N-(2-
hydroxyethyl)-1H-benzimidazole-5-carboxamide, 6-(5-Methyl-1H-benzimidazol-1-
yl)-N-[(1S)-1-phenylethyl]pyrazin-2-amine, N-[1-(6-{[(1S)-1-phenylethyl] amino}
pyrazin-2-yl)-1H-benzimidazol-6-yl]nicotinamide, N-methyl-1-(6-{[(1S)-1-
phenylethyl]amino} pyrazin-2-yl)-1H-benzimidazole-5-carboxamide, N-[ 1-(6-{[(1S)-
1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-6-yl]-2,2-
dimethylpropanamide, N-methyl-1-(6-{[(1S)-1-phenylethyl]amino} pyrazin-2-yl)-1H-
benzimidazole-6-carboxamide, N-[1-(6- {[(1S)-1 -phenylethyl] amino }pyrazin-2-yl)-
1 H-benzimidazol-5 -yl] -2,2-dimethylpropanamide, 1-(6-{[(15)-1 -Phenylethyl] amino}
pyrazin-2-yl)-1H-benzimidazol-5-amine, 2-Methoxy-N-[1-(6-{[(1S)-1-phenylethyl]
amino}pyrazin-2-yl)-1H-benzimidazol-5-yl]acetamide, 1-(6-{[(1S)-1-phenylethyl]
amino}pyrazin-2-yl)-1H-benzimidazol-6-amine, 2-Methoxy-N-[ 1-(6- {[(1S)-1-
phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-6-yl]acetamide, N-Ben2yl-1-[6-
([(1S)-1 -phenylethyl] amino)pyrazin-2-yl] -1H-benzimidazole-5 -carboxamide, N- [1-
(6- {[(1S)-1 -phenylethyl]amino} pyrazin-2-yl)-1H-benzimidazol-5-yl] pyrazine-2-
carboxamide, 1-(6-{[(1S)-1-Phenylethyl]amino}pyrazin-2-yl)-N-phenyl-1H-
benzimidazole-5 -carboxamide, N-[1-(6-{[(15)-1-phenylethyl] amino} pyrazin-2-yl)-
1H-benzimidazol-6-yl] pyrazine-2-carboxamide, N- [ 1 -(6- {[(1S)-1-phenylethyl]
amino } pyrazin-2-yl)-1H-benzimidazol-6-yl] acetamide, 6-{5-[(4-Methylpiperazin-1-
yl)methyl]-1H-benzimidazol-1-yl} -N- [(1S)-1- phenylethyl]pyrazin-2-amine, N- [1-(6-
{[(1S)-1-Phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-5-yl]acetamide, [1-(6-
{[(1S)-1-Phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-5-yl]methanol, N-[l-(6-
{[(1S)-1-phenylethyl]amino} pyrazin-2-yl)-1H-benzimidazol-6-yl]benzamide, [1-(6-
{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-6-yl]methanol, N-[l-(6-
{[(1S)-1 -phenylethyl] amino} pyrazin-2-yl)-1 H-benzimidazol-5 -yl]benzamide, 1-(6-
{[(1S)-1 -phenylethyl] amino} pyrazin-2-yl)-N- [2-(dimethylamino)ethyl] -1H-
benzimidazole-5-carboxamide, 1-[6-{[(1S)-1-Phenylethyl]amino}pyrazin-2-yl]-N-
(pyridin-3-ylmethyl)-1H-benzimidazol-5-amine, tert-butyl (2S)-2-({[1-(6-{[(1S)-1 -
phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-5-yl]amino} carbonyl)
pyrrolidine-1-carboxylate, 6-(3H-imidazo[4,5-c]pyridin-3-yl)-N-[(15)-1-phenylethyl]
pyrazin-2-amine, 6-( 1 H-benzimidazol-1 -yl)-N- [1-(4-fluorophenyl)ethyl]pyrazin-2-
amine, 6-(lH-imidazo[4,5-c]pyridin-1-yl)-N-[(1S)-1-phenylethyl]pyrazin-2-amine,
6-( 1 H-benzimidazol-1 -yl)-N-[(1S)-1-(4-pyridin-3-ylphenyl)ethyl]pyrazin-2-amine,
(2S)-N-[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-1H-benzimidazol-5-yl]
pyrrolidine-2-carboxamide, N- [(1S)-1 -phenylethyl] -6-(5-pyridin-4-yl-1H-
benzimidazol-1-yl)pyrazin-2-amine, N-[(1S)-1-phenylethyl]-6-(5-pyridin-3-yl-1H-
benzimidazol-1 -yl)pyrazin-2-amine, 6-(5 -bromo-1H-benzimidazol-1-yl)-N- [(1S)-1-
phenylethyl]pyrazin-2-amine, N-[3-(1H-imidazol-1-yl)propyl]-1-[6-([(1S)-1-
phenylethyl]amino)pyrazin-2-yl] -1H-benzimidazole-6-carboxamide, N-1H-
benzimidazole-6-carboxamide, N-(3-morpholin-4-ylpropyl)-1-[6-([(1S)-1-
phenylethyl] amino)pyrazin-2-yl] -1H-benzimida2;ole-6-carboxamide, N-(3-
morpholin-4-ylpropyl)-1-[6-([(1S)-1-phenylethyl]amino)pyrazin-2-yl]-1H-
benzimidazole-5-carboxamide, N-[1-(6-{[(1S)-1-phenylethyl]amino}pyrazin-2-yl)-
1H-benzimidazol-5-yl]piperidine-3-carboxamide, 6-(1H-benzimidazol-1-yl)-N-
[(1S)-1-pyridin-3-ylethyl]pyrazin-2-amine, 6-(1H-benzimidazol-1-yl)-N-[(1S)-1-
(1,1"-biphenyL-4-yl)ethyl]pyrazin-2-amine N-[1-(6- {[(1S)-1-phenylethyl] aminq}
pyrazin-2-yl)-1H-benzimidazol-5-yl]benzenesulfonamide and 6-( 1H-benzimidazol-
1 -yl)-N-[(1S)-1-(1,1" -biphenyl-4-yl)ethyl]pyrazin-2-amine.
10. A compound wherein the compound is selected from the group consisting of:
11. A composition comprising a carrier and at least one compound as claimed in any one
of claims 1 to 10.
12. A pharmaceutical composition comprising at least one of the compounds as claimed
in any one of claims 1 to 10 capable of treating a protein kinase-associated disorder
in an amount effective therefor, and a pharmaceutically acceptable vehicle or diluent.
13. A composition as claimed in claim 11 or claim 12 for treating a protein kinase
associated disease state.
14. A composition as claimed in claim 13, wherein the disease state involves a receptor
tyrosine kinase selected from the group consisting of EGF, HER2, HER3, HER4, IR,
IGF-1R, IRR, PDGFR.alpha., PDGFR.beta., CSFIR, C-Kit, C-fms,Flk-1R, Flk4,
KDR/Flk-1, Flt-1, FGFR-1R, FGFR-2R, FGFR-3R and FGFR-4R.
15. A composition as claimed in claim 13, wherein the disease state involves a cellular
tyrosine kinase selected from the group consisting of Src, Frk, Btk, Csk, Abl,
ZAP70, Fes/Fps, Fak, Ack, Yes, Fyn, Lyn, Lck, Blk, Hck, Fgr and Yrk.
16. A composition as claimed in claim 13, wherein the disease state involves a tyrosine
kinase selected from the group consisting of JAK1, JAK2, JAK3 and TYK2.
17. A composition as claimed in claim 13, wherein the disease state involves a
serine/threonine kinase selected from the group consisting of ERK2, c-Jun, p38
MAPK, PKA, PKB, PKC, a cyclin-dependent kinase, CDK1, CDK2, CDK3, CDK4,
CDK5, CDK6, CDK7, CDK8, CDK9, CDK10, and CDK11.
18. A composition as claimed in claim 13, wherein the disease state is selected from the
group consisting of Atopy, such as Allergic Asthma, Atopic Dermatitis (Eczema),
and Allergic Rhinitis; Cell Mediated Hypersensitivity, such as Allergic Contact
Dermatitis and Hypersensitivity Pneumonitis; Rheumatic Diseases, such as Systemic
Lupus Erythematosus (SLE), Rheumatoid Arthritis, Juvenile Arthritis, Sjogren"s
Syndrome, Scleroderma, Polymyositis, Ankylosing Spondylitis, 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, Varicella-Zoster Virus (VZV), Human Papilloma
Virus (HPV), Cancer, such as Leukemia, Lymphoma and Prostate Cancer.
A compound of the general formula (I) or pharmaceutically acceptable salts, hydrates, solvates, crystal forms or
diastereomers thereof is described. A method of treating protein kinase-associated disease states using the compound of formula (I)
is also described.

Documents:

1743-kolnp-2004-granted-abstract.pdf

1743-kolnp-2004-granted-assignment.pdf

1743-kolnp-2004-granted-claims.pdf

1743-kolnp-2004-granted-correspondence.pdf

1743-kolnp-2004-granted-description (complete).pdf

1743-kolnp-2004-granted-drawings.pdf

1743-kolnp-2004-granted-examination report.pdf

1743-kolnp-2004-granted-form 1.pdf

1743-kolnp-2004-granted-form 18.pdf

1743-kolnp-2004-granted-form 3.pdf

1743-kolnp-2004-granted-form 5.pdf

1743-kolnp-2004-granted-letter patent.pdf

1743-kolnp-2004-granted-others.pdf

1743-kolnp-2004-granted-pa.pdf

1743-kolnp-2004-granted-reply to examination report.pdf

1743-kolnp-2004-granted-specification.pdf


Patent Number 214081
Indian Patent Application Number 01743/KOLNP/2004
PG Journal Number 05/2008
Publication Date 01-Feb-2008
Grant Date 30-Jan-2008
Date of Filing 17-Nov-2004
Name of Patentee CYTOPIA PTY LTD.
Applicant Address LEVEL 5, BAKER HEART RE4SEARCH INSTITUTE BUILDING COMMERCIAL ROAD MELBOOURNE VICTORIA AUSTRALIA.
Inventors:
# Inventor's Name Inventor's Address
1 WILKS ANDREW FREDERICK 6 MACFARLAN4E LANE SOUTH YARRA VICTORIA AUSTRALIA
2 BURNS CHRISTOPHER JOHN 3 BRONING SDTREET SEDDON VICTORIA AUSTRALIA
3 BU XIANYONG 1 DAIRY COURT ROSANNA EAST VICTORIA AUSTRALIA
PCT International Classification Number C07D403/04
PCT International Application Number PCT/AU03/00628
PCT International Filing date 2003-05-23
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 60/398,998 2002-07-26 Australia
2 60/398,998 2002-07-26 Australia