HK1125631B - Benzazole derivatives, compositions, and methods of use as aurora kinase inhibitors - Google Patents

Description

Benzoxazole-based derivatives as Aurora kinase inhibitors, compositions, and methods of use

CROSS-REFERENCE TO RELATED APPLICATIONS

Priority of U.S. provisional application No.60/772,497 entitled "benzoxazole series derivatives as Aurora kinase inhibitors, compositions and methods of use" filed 2006, month 2, 10 and application No.60/791,187 entitled "benzoxazole series derivatives as Aurora kinase inhibitors, compositions and methods of use" filed 2006, month 4, 11 and application No. 35 u.s.c. § 119(e), the entire contents of which are incorporated herein by reference.

Technical Field

The present invention relates to azole derivatives useful as Aurora kinase inhibitors and methods of treating cancer using benzoxazole derivatives.

Background

A better understanding of the signal transduction pathways and enzymes underlying disease etiology and pathophysiology greatly facilitates the search for new therapeutic agents. One important class of enzymes that has been the subject of extensive research targeting disease processes are protein kinases.

Protein kinases are key regulators of cell growth, differentiation, metabolism and function. Protein kinases are a family of structurally related enzymes that are responsible for controlling a variety of signal transduction processes within cells (The Protein Kinase enzymes Book, I and II, Academic Press, san diego, calif.: 1995). Almost all protein kinases contain a catalytic domain consisting of about 250 to 300 amino acids. In general, protein kinases mediate their intracellular signaling through the catalytic transfer of gamma-phosphoryl from ATP to target protein substrates. Protein kinases are classified into several families according to the substrates they phosphorylate. Sequence motifs corresponding to each of these kinase families have been identified, such as protein-tyrosine, protein-serine/threonine, and lipids (Hanks, s.k., Hunter, t., FASEB j.1995, 9576-. Protein kinases allow cells to make decisions by acting as molecular "switches" that can perturb or modulate the function of target proteins in response to a variety of stimuli.

Aberrant protein kinase-mediated signaling in cells is the underlying cause of many pathophysiological states. These disease states include, but are not limited to, autoimmune diseases, allergy and asthma diseases, neurological and neurodegenerative diseases, metabolic diseases, alzheimer's disease, cardiovascular diseases and cancer. Protein kinases are therefore considered as rational drug targets for therapeutic intervention, and protein kinase inhibitors are considered as effective therapeutic agents.

The Aurora family of serine/threonine protein kinases are essential for Cell proliferation (Trends Cell Biology 9, 454-459 (1999); nat. Rev. mol. Cell Biology 2, 21-32 (2001); Trends in Cell Biology 11, 49-54 (2001)). The human Aurora kinase family consists of three highly homologous kinases (a or "2", B or "1" and C or "3"). During normal cell proliferation, these kinases are involved in chromosome segregation, mitotic spindle function, and cytoplasmic changes. Aurora kinase expression is low in resting cells, peaking during G2 and mitosis of the cell cycle. Several proposed mammalian AurorｃA kinase substrates include histone H3, TPX2, myosin II regulatory light chain, CENP- ｃA, and protein phosphatase 1, which are important sites for cell division.

Since their key roles in mitotic processes and cell division have been elucidated, Aurora kinases have been implicated in tumorigenesis. For example, Aurora kinase gene amplification and overexpression have been reported in many cancers. Coding Single Nucleotide Polymorphisms (SNPs) have been identified, which are significantly more frequent in late-stage gastric Cancer relative to early-stage gastric Cancer, and are associated with elevated kinase activity (Cancer lett. jan 10, 2006). Overexpression of Aurora A includes centrosome expansion, aneuploidy and transformation of rodent fibroblasts (Bischoff, J.R.et al.EMBO.J 17, 3052-3065 (1998); nat. Genet. Oct 20 (2): 189-93 (1998)). This oncogenic activity is likely due to the development of chromosomal instability. In fact, there is a strong correlation between Aurora A overexpression and chromosomal aneuploidy in breast and gastric cancers (int., J. cancer 92, 370-. Aurora B expression is elevated in cell lines derived from colon, breast, lung, melanoma, kidney, ovary, pancreas, CNS, gut and leukemic tumors (Oncol Res.2005; 15 (1): 49-57; Tatsuka et al 1998, 58, 4811-. In Prostate cancer, increased nuclear expression of Aurora B is observed in high Gleason grade degenerative Prostate cancer tissues relative to mid-low levels, with phosphorylation of histone H3 substrate (protate 66 (3): 326-33 (2003)). Aurora C is overexpressed in primary colorectal cancers (Journal of Biological Chemistry 274, 7334-7340 (1999); Jpn. J. cancer Res.91, 1007-1014 (2000)).

Because Aurora kinase inhibition in tumor cells can lead to mitotic arrest and apoptosis, these kinases are important targets for cancer therapy. Due to the central role of mitosis in the progression of virtually all malignancies, Aurora kinase inhibitors are therefore expected to have the potential to block cancer or tumor growth, with broad application in human cancers or tumors.

Summary of The Invention

The present invention provides substituted benzoxazole-based derivatives and compositions that inhibit Aurora kinase. In one embodiment, the invention provides a compound of formula (I) as depicted below. In another embodiment, the present invention provides a process for the preparation of a compound of formula (I). In another embodiment, the present invention provides a pharmaceutical composition comprising a compound of formula (I). In another embodiment, the invention provides methods of treating human or animal disorders (disorders) using compounds of formula (I) and pharmaceutical compositions comprising compounds of formula (I). The compounds of the invention are useful as inhibitors of Aurora kinases and thus may be useful in the management, treatment, control and adjunct treatment of diseases mediated by Aurora kinase activity, such as cell proliferative disorders, including cancer.

Brief description of the drawings

FIG. 1 shows the MiaPaCa-2 tumor growth curves, wherein a represents the vector of example 88 and erlotinib (erlotinib); o represents erlotinib at a dose of 50mg/kg daily for 14 days p.o. day; □ represents example 88 at a dose of 10mg/kg daily, i.p., b.i.d. for 10 days; ● represents example 88 with erlotinib.

FIG. 2 shows the MiaPaCa-2 tumor growth curve, where ■ represents the vector of example 88 and gemcitabine (gemcitabine); a dose of 10mg/kg daily, i.p., b.i.d. for 10 days, representing example 88; □ represents gemcitabine, at a dose of 120mg/kg, i.p., q3d × 4 per day; the value of "good" represents the value of example 88 and gemcitabine.

FIG. 3 shows BT-474 tumor growth curves in athymic SCID mice, where ■ represents the vector of example 88; o represents trastuzumab (trastuzumab) at a dose of 10mg/kg twice a week for up to 4 weeks i.p.; a represents example 88 at a dose of 30mg/kg daily, i.p., b.i.d. for 3 days, then discontinued for 2 days for a total of 5 cycles; □ represents example 89 with trastuzumab.

Detailed description of the invention

The present invention provides compounds that inhibit Aurora kinase. These compounds are useful for inhibiting Aurora kinases in vitro, and can be used to treat cell proliferative disorders, including cancer, in a patient.

In one aspect, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt or prodrug thereof:

wherein

X is-NH-, -O-or-S-,

e is-CH₂-, -NH-, -O-or-S-,

G¹and G²Independently selected from the group consisting of: aryl, heteroaryl, fused arylcycloalkyl, fused cycloalkylaryl, fused cycloalkylheteroaryl, fused heterocyclylaryl and fused heterocyclylheteroaryl, wherein G¹And G²Optionally independently selected from R^b1 to 7 times;

L¹selected from the group consisting of: direct bond, -CH₂-、-O-、-O-CH₂-、-CH₂-O-、-N(R⁶)-、-C(O)-、-C(O)N(R⁶)-、-N(R⁶)C(O)-、-N(R⁶)C(O)N(R⁷)-、-N(R⁶)C(O)O-、-OC(O)N(R⁶)-、-N(R⁶)SO₂-、-SO₂N(R⁶)-、-C(O)-O-、-O-C(O)-、-S-、-S(O)-、-S(O)₂-、-N(R⁶)SO₂N(R⁷)-、-N＝N-、-C(R⁸)＝C(R⁹) -and-C ≡ C-,

wherein

R⁶And R⁷Independently selected from R^dAnd R^eA group of (a); and

R⁸and R⁹Independently selected from R^eAnd R^fA group of (a);

a is a direct bond or a group-L²-Y-L³-, wherein

L²And L³Independently selected from the group consisting of:

the direct bond of the compound has a direct bond,

-C_1-10an alkylene group or a substituted alkylene group,

-C_2-10an alkenylene group, a carboxyl group,

-C_2-10an alkynylene group which is a substituent of a heterocyclic ring,

an arylene group, a halogenated arylene group,

-a heteroarylene group,

-a cycloalkylene group,

-a heterocyclylene group,

wherein the carbon atoms of the alkylene, alkenylene, alkynylene, arylene, heteroarylene, cycloalkylene, and heterocyclylene group are optionally independently selected from R^cThe substituent group(s) is substituted for 1 to 4 times;

y is a direct bond, -O-, -N (R)¹⁰)、-S-、SO₂-、-C(O)N(R¹⁰)-、-N(R¹⁰)-C(O)-、-N(R¹¹)C(O)N(R¹⁰)-、-N(R¹⁰)SO₂-、-SO₂N(R¹⁰)-、-C(O)-O-、-N(R¹¹)SO₂N(R¹⁰) -, -O-CO-or-N-,

wherein

R¹⁰And R¹¹Independently selected from the group consisting of: r^dAnd R^eAnd are and

q is selected from the group consisting of:

and

wherein R is¹⁶And R¹⁷Independently selected from the group consisting of: r^dAnd R^e；

2) -a heteroaryl group;

-a heterocyclic group;

-a fused cycloalkylheteroaryl group;

-a fused heterocyclylaryl group;

-a fused heterocyclylheteroaryl group;

-a fused aryl heterocyclyl group;

-a fused heteroaryl cycloalkyl group; and

-a fused heteroaryl heterocyclyl group;

wherein the heteroaryl, heterocyclyl, fused cycloalkylheteroaryl, fused heterocyclylaryl, fused heterocyclylheteroaryl, fused arylheterocyclyl, fused heteroarylcycloalkyl, and fused heteroarylheterocyclyl are optionally independently selected from R^cThe substituent group(s) is substituted for 1 to 4 times; and

3) a ring system comprising at least one nitrogen atom selected from the group consisting of:

and

wherein

n, m, p, q and r are independently 0 to 4, such that n + m + p equals 2 to 5 and q + r equals 2 to 5, the cycloalkyl or heterocyclic ring system optionally being in (CH)₂) By R on a carbon atom¹⁸Or R¹⁹Substituted 1-2 times, wherein R¹⁸And R¹⁹Independently selected from R^fAnd R^gA group of components selected from the group consisting of,

J¹selected from the group consisting of:

and

J³and J⁵Independently selected from the group consisting of: -CH₂-、-O-、-S-、-S(O)₂-、-C(O)-、-C(O)N(H)-、-NHC(O)-、-NHC(O)N(H)-、-NHSO₂-、-SO₂N(H)-、-C(O)-O-、-O-C(O)-、-NHSO₂NH-，

And

R²⁹and R³⁰Independently selected from R^dAnd R^eA group of (a);

R³¹is R^f；

R¹Is R^b；

R^bIs that

a) -a cycloalkyl group,

b) -a cyano group,

c)-OR^d，

d)-NO₂，

e) -a halogen, in the form of a halogen,

f) -a halogenated alkyl group,

g)-S(O)_sR^d，

h)-SR^d，

i)-S(O)₂OR^d，

j)-S(O)_sNR^dR^e，

k)-NR^dR^e，

l)-O(CR^fR^g)_tNR^dR^e，

m)-C(O)R^d，

n)-CO₂R^d，

o)-CO₂(CR^fR^g)_tC(O)NR^dR^e，

p)-OC(O)R^d，

q)-C(O)NR^dR^e，

r)-NR^dC(O)R^e，

s)-OC(O)NR^dR^e，

t)-NR^dC(O)OR^e，

u)-NR^dC(O)NR^dR^e，

v)-CF₃，

w)-OCF₃

x)-C_1-10an alkyl group, a carboxyl group,

y)-C_2-10an alkenyl group, which is a radical of an alkenyl group,

z)-C_2-10an alkynyl group,

aa)-C_1-10an alkylene-aryl group, a substituted alkylene-aryl group,

bb)-C_1-10alkylene-heteroaryl, or

cc) -a heteroaryl group, and (b) a pharmaceutically acceptable salt thereof,

wherein alkyl, alkenyl, alkynyl, aryl, heteroaryl and cycloalkyl are optionally independently selected from R^cThe substituent group(s) is substituted for 1 to 4 times;

R^cis that

a) -a halogen, in the form of a halogen,

b) -an amino group,

c) -a carboxyl group,

d)-C_1-4an alkyl group, a carboxyl group,

e)-O-C_1-4an alkyl group, a carboxyl group,

f) -a cycloalkyl group,

g) -an O-cycloalkyl group, a cycloalkyl group,

h) -an aryl group,

i)-C_1-4an alkylene-aryl group, a substituted alkylene-aryl group,

j) -a hydroxyl group,

k)-CF₃，

l) -O-aryl, or a pharmaceutically acceptable salt thereof,

m) -a heteroaryl group, a pharmaceutically acceptable salt thereof,

n) -heteroaryl-C_1-10An alkyl group, a carboxyl group,

o) a heterocyclic group,

p)-CO₂-C_1-10alkyl, or

q)-CO₂-C_1-10An alkyl-aryl group, which is a cyclic alkyl group,

R^dand R^eIndependently selected from hydrogen, C_1-10Alkyl radical, C_2-10Alkenyl radical, C_2-10Alkynyl, cycloalkyl, -C_1-10Alkylene-cycloalkyl, aryl, heterocyclyl, whereinAlkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocyclyl are optionally substituted with 1-4 substituents independently selected from R^cSubstituted with the substituent(s); or R^dAnd R^eTogether with the atoms to which they are attached form a 5-7 membered heterocyclic ring containing 0-2 additional heteroatoms independently selected from oxygen, sulfur and nitrogen, and optionally substituted with R^cThe substitution is carried out for 1 to 3 times,

R^fand R^gIndependently selected from hydrogen, C_1-10Alkyl, cycloalkyl, -C_1-10Alkylene-cycloalkyl and aryl, wherein alkyl, cycloalkyl and aryl are optionally substituted with 1-4 substituents independently selected from R^cSubstituted with the substituent(s); or R^fAnd R^gTogether with the carbon to which they are attached form a 5-7 membered ring containing 0-2 heteroatoms independently selected from oxygen, sulfur and nitrogen, and optionally substituted with R^cThe substitution is carried out for 1 to 3 times,

s is an integer of 1 to 2,

t is an integer of 1 to 10,

u is an integer of 0 to 1,

v is an integer of 0 to 2.

In one embodiment, X is-NH-.

In another embodiment, X is-NH-and E is-NH-.

In one embodiment of the compounds of formula (I), G²Selected from the group consisting of: phenyl, naphthyl, isoquinolin-3-yl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, thiophen-2-yl, thiazol-2-yl, imidazol-2-yl, benzothiazol-2-yl, and 4, 5, 6, 7-tetrahydro-thiazolo [5, 4-c]-a pyridin-2-yl group,

wherein G is²Optionally is selected from R^bThe substituent(s) is substituted 1 to 4 times.

In a further embodiment, G²Substituted with at least one substituent selected from the group consisting of: halo, phenyl, C_1-10Alkyl, piperazin-1-yl, 4- (C)_1-10Alkyl) -piperazinesOxazin-1-yl, C_1-10Alkoxy, haloalkyl, cycloalkyl and C_1-10Alkylene-cycloalkyl groups.

In a further embodiment, G²Is phenyl, naphthyl, isoquinolin-3-yl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, thiophen-2-yl, thiazol-2-yl, imidazol-2-yl, benzothiazol-2-yl, wherein G²Is unsubstituted or substituted with at least one substituent selected from the group consisting of: chloro, fluoro, methyl, ethyl, propyl, isopropyl, tert-butyl, phenyl, methoxy, trifluoromethyl, trifluoromethoxy, and cyclopentyl.

In one embodiment of the compounds of formula (I), L¹is-C (O) -NH-or-NH-C (O) -.

In another embodiment of the compounds of formula (I), L¹is-C (R)⁸)＝C(R⁹)-。

In another embodiment of the compounds of formula (I), G¹Selected from the group consisting of:

phenyl, phenyl,

Pyrazol-3-yl,

Benzothiazol-5-yl, benzothiazol-6-yl, benzothiazol,

Benzimidazol-5-yl, benzimidazol-6-yl,

Benzoxazol-5-yl, benzoxazol-6-yl,

Benzotriazol-5-yl, benzotriazol-6-yl,

Benzisoxazol-5-yl, benzisoxazol-6-yl,

Indol-5-yl, indol-6-yl,

2H-indazol-6-yl,

1H-indazol-3-yl, 1H-indazol-4-yl, 1H-indazol-5-yl, 1H-indazol-6-yl,

Quinolin-6-yl, quinolin-7-yl,

Quinazolin-4-yl,

2-oxindol-5-yl, 2-oxindol-6-yl,

2- (1H) -benzimidazolon-5-yl,

3-indazololin-5-yl and 3-indazololin-6-yl,

wherein G is¹Optionally is selected from R^bThe substituent(s) is substituted 1 to 4 times.

In a further embodiment, G¹Is unsubstituted or substituted by at least one group selected from: halo, phenyl, C_1-10Alkyl, piperazin-1-yl, 4- (C)_1-10Alkyl) -piperazin-1-yl, -C_1-10Alkoxy, -C_1-10alkylene-OH, -haloalkyl, -cycloalkyl, -C_1-10Alkylene-cycloalkyl, morpholin-4-yl, -C_1-10-alkylene-morpholin-4-yl, pyrrol-1-yl, -amino, -NH- (C)_1-10Alkyl), -N (C)_1-10Alkyl radical)₂、-NHC(O)-C_1-10Alkyl, -NHC (O) - (1- (C)_1-10Alkyl) -piperidin-4-yl), -NHC (O) -phenyl, -NH-C_1-10Alkylene-morpholin-4-yl, -O-C_1-10Alkylene-morpholin-4-yl and-NH-C_1-10alkylene-OH.

In a further embodiment, L¹is-NHC (O) -or-C (O) -NH-, G¹Is 1H-indazol-5-yl or 1H-indazol-6-yl, wherein G¹Optionally is selected from R^bThe substituent(s) is substituted 1 to 4 times.

In a further embodiment, G¹Is 1H-indazol-5-yl or 1H-indazol-6-yl, wherein G¹Is unsubstituted or substituted in the 3-position with a substituent selected from the group consisting of: halo, phenyl, C_1-10Alkyl, piperazin-1-yl, 4- (C)_1-10Alkyl) -piperazin-1-yl, -C_1-10Alkoxy, -C_1-10alkylene-OH, -haloalkyl, -cycloalkyl, -C_1-10Alkylene-cycloalkyl, morpholin-4-yl, -C_1-10-alkylene-morpholin-4-yl, pyrrole-1-yl, -amino, -NH- (C)_1-10Alkyl), -N (C)_1-10Alkyl radical)₂、-NHC(O)-C_1-10Alkyl, -NHC (O) - (1- (C)_1-10Alkyl) -piperidin-4-yl), -NHC (O) -phenyl, -NH-C_1-10Alkylene-morpholin-4-yl, -O-C_1-10Alkylene-morpholin-4-yl and-NH-C_1-10alkylene-OH.

In another embodiment, u is 1, A is a direct bond, and Q is selected from the group consisting of:

4-(C_1-10alkyl) -piperazin-1-yl group,

piperidin-1-yl (piperadine-1-yl),

a group of morpholine-4-yl and a group of morpholine-4-yl,

-NH-C_1-10an alkyl group, a carboxyl group,

-N-(C_1-10alkyl radical)₂，

-N-(C_1-10Alkyl) (cycloalkyl), and

-NH-cycloalkyl.

In one embodiment, u is 0 and v is 0. In another embodiment, u is 1 and v is 0. In another embodiment, u is 0 and v is 1.

In another embodiment, u is 0 and v is 1, R¹Selected from the group consisting of:

-C_1-10an alkyl group, a carboxyl group,

-a cycloalkyl group,

-C_1-10an alkylene-cycloalkyl group, which is,

-C_1-10a halogenated alkyl group,

-a phenyl group,

-O-C_1-10an alkyl group, a carboxyl group,

-O-cycloalkyl, and

-O-C_1-10a haloalkyl group.

In another embodiment, X is-NH-, E is-NH-, v is 0, and L¹is-NHC (O) -or-C (O) NH-, G¹Is 1H-indazol-6-yl or 1H-indazol-5-yl, wherein G¹Optionally is selected from R^bThe substituent(s) is substituted 1 to 4 times. In another embodiment, G¹Is unsubstituted.

In another embodiment, the compound of formula (I) has the formula:

wherein

G¹、G²、L¹Q and A are as defined above.

In another embodiment, the compound of formula (I) has the formula:

wherein

G¹、G²And Q is as defined above.

In further embodiments of formula (Ib), Q is selected from the group consisting of: 4- (C)_1-10Alkyl) -piperazin-1-yl, piperidin-1-yl, morpholin-4-yl, -NH-C_1-10Alkyl, -N- (C)_1-10Alkyl radical)₂、-N-(C_1-10Alkyl) (cycloalkyl) and-NH-cycloalkyl.

In further embodiments of formula (Ib), Q is selected from the group consisting of: morpholin-4-yl, 4-methyl-piperazin-1-yl, diethylamino, 2, 6-dimethylmorpholin-4-yl, (2-dimethylaminoethyl) -methylamino, 4-dimethylaminopiperidin-1-yl, dipropylamino, bis- (2-methoxyethyl) amino, 4-hydroxypiperidin-1-yl, ethyl- (2-methoxyethyl) amino, pyrrolidin-1-yl, N-ethyl-N' - (2-methoxyethyl) amino, ethylpropylamino, 4-isopropylpiperazin-1-yl and ethylmethylamino.

In further embodiments of formula (Ib), G²Selected from the group consisting of: phenyl, naphthyl, isoquinolin-3-yl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, thiophen-2-yl, thiazol-2-yl, imidazol-2-yl, benzothiazol-2-yl, and 4, 5, 6, 7-tetrahydro-thiazolo [5, 4-c]-pyridin-2-yl, wherein G²Optionally is selected from R^bThe substituent(s) is substituted 1 to 4 times.

In further embodiments of formula (Ib), G²Selected from the group consisting of: phenyl and pyridin-2-yl, in which G²Is unsubstituted or substituted with at least one substituent selected from the group consisting of: methyl, methoxy, trifluoromethyl and trifluoromethoxy.

In further embodiments of formula (Ib), G¹Selected from the group consisting of: phenyl, pyrazol-3-yl, benzothiazol-5-yl, benzothiazol-6-yl, benzimidazol-5-yl, benzimidazol-6-yl, benzoxazol-5-yl, benzoxazol-6-yl, benzotriazol-5-yl, benzotriazol-6-yl, benzisoxazol-5-yl, benzisoxazol-6-yl, indol-5-yl, indol-6-yl, 2H-indazol-6-yl, 1H-indazol-3-yl, 1H-indazol-4-yl, 1H-indazol-5-yl, 1H-indazol-6-yl, quinolin-6-yl, benzothiazol-6-yl, benzoimidazol-5-yl, benzoimidazol-6-yl, benzoisoxazol-6-yl, benzoiso, Quinolin-7-yl, quinazolin-4-yl, 2-oxindol-5-yl, 2-oxindol-6-yl, 2- (1H) -benzimidazolon-5-yl, 3-indazolon-5-yl, and 3-indazolon-6-yl, wherein G¹Optionally is selected from R^bThe substituent(s) is substituted 1 to 4 times.

In another embodiment, the compound of formula (I) has the formula:

wherein G is¹、G²And R¹Is as defined above。

In a further embodiment of the compounds of formula (Ic), R¹Selected from the group consisting of: -C_1-10Alkyl, -cycloalkyl, -C_1-10Alkylene-cycloalkyl, -C_1-10Haloalkyl, -phenyl, -O-C_1-10Alkyl, -O-cycloalkyl and-O-C_1-10A haloalkyl group.

In a further embodiment of formula (Ic), G²Selected from the group consisting of: phenyl, naphthyl, isoquinolin-3-yl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, thiophen-2-yl, thiazol-2-yl, imidazol-2-yl, benzothiazol-2-yl, and 4, 5, 6, 7-tetrahydro-thiazolo [5, 4-c]-pyridin-2-yl, wherein G²Optionally is selected from R^bThe substituent(s) is substituted 1 to 4 times.

In further embodiments of formula (Id), G¹Selected from the group consisting of: phenyl, pyrazol-3-yl, benzothiazol-5-yl, benzothiazol-6-yl, benzimidazol-5-yl, benzimidazol-6-yl, benzoxazol-5-yl, benzoxazol-6-yl, benzotriazol-5-yl, benzotriazol-6-yl, benzisoxazol-5-yl, benzisoxazol-6-yl, indol-5-yl, indol-6-yl, 2H-indazol-6-yl, 1H-indazol-3-yl, 1H-indazol-4-yl, 1H-indazol-5-yl, 1H-indazol-6-yl, quinolin-6-yl, benzothiazol-6-yl, benzoimidazol-5-yl, benzoimidazol-6-yl, benzoisoxazol-6-yl, benzoiso, Quinolin-7-yl, quinazolin-4-yl, 2-oxindol-5-yl, 2-oxindol-6-yl, 2- (1H) -benzimidazolon-5-yl, 3-indazolon-5-yl, and 3-indazolon-6-yl, wherein G¹Optionally is selected from R^bThe substituent(s) is substituted 1 to 4 times.

In the compounds of formula (I), the various represented functional groups should be understood as having a point of attachment at the functional group with a hyphen. In other words at-C_1-10In the case of alkylene-aryl, it is to be understood that the point of attachment is alkylene; an example would be benzyl. In a reaction zone such as-C (O) -NH-C_1-10In the case of alkylene-aryl groups and the like, the point of attachment is the carbonyl carbon.

The term "Aurora kinase inhibitor" or "inhibitor of Aurora kinase" is used to characterize a compound having the structure defined herein, which is capable of acting on an Aurora kinase, inhibiting its enzymatic activity. Inhibition of Aurora kinase enzyme activity indicates a reduction in the ability of Aurora kinase to phosphorylate substrate peptides or proteins. In various embodiments, Aurora kinase activity is reduced by at least about 50%, at least about 75%, at least about 90%, at least about 95%, or at least about 99%. In various embodiments, the concentration of Aurora kinase inhibitor required to reduce Aurora kinase enzyme activity is less than about 1 μ M, less than about 500nM, or less than about 100 nM.

In some embodiments, such inhibition is selective, that is, the Aurora kinase inhibitor reduces the ability of the Aurora kinase to phosphorylate a substrate peptide or protein at a concentration that is lower than the concentration of the inhibitor required to produce another unrelated biological effect, such as reducing the enzymatic activity of a different kinase.

The term "comprising" as used herein means "including but not limited to".

Also included within the scope of the present invention are the individual enantiomers of the compounds represented by formula (I) above as well as any mixtures thereof, whether fully or partially racemic. The present invention also encompasses mixtures of individual enantiomers of compounds represented by formula (I) above and diastereomers thereof, wherein one or more stereocenters are inverted. Unless otherwise specified, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, having the structure except that hydrogen atoms are replaced by deuterium or tritium or carbon atoms are replaced by deuterium¹³C-or¹⁴Compounds other than C-rich carbon substitutions fall within the scope of the present invention.

In another aspect, the invention provides a pharmaceutically acceptable salt, solvate or prodrug of a compound of formula (I). In one embodiment, the prodrug comprises a biohydrolyzable ester or biohydrolyzable amide of the compound of formula (I).

Examples of compounds of formula (I) of the present invention having potentially useful biological activity are listed in table 1 below. The Compounds of formula (I) inhibit Aurora kinase activity using the peptide phosphorylation assay described in example 102The ability to characterize was confirmed by representative compounds of formula (I) as listed in Table 1. IC for inhibition of Aurora kinase by the compounds of formula (I) in Table 1₅₀May be less than or equal to 1 micromolar (μ M; 10)^-6M)。

Compounds that inhibit Aurora kinase activity are potentially useful for treating cell proliferative disorders. The compounds of formula (I) of the present invention may therefore be particularly useful in the treatment of cancer.

TABLE 1

Examples	Name (R)
		1	2- (isoquinolin-3-ylamino) -1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide
2	2- (isoquinolin-3-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-5-yl) -amide
		3	2- (pyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-5-yl) -amide
4	2- (pyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (2-methyl-benzoxazol-5-yl) -amide
		5	2- (pyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
6	2- (pyridin-3-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		7	2- (pyridin-2-ylamino) -1H-benzimidazole-4-carboxylic acid benzothiazol-6-ylamide
8	2- (pyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-benzotriazol-5-yl) -amide
		9	2- (2, 4-dichlorophenylamino) -3H-benzimidazole-5-carboxylic acid (1-methyl-1H-indazol-5-yl) -amide
10	2- (2, 4-dichlorophenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-5-yl) -amide
		11	2-phenylamino-1H-benzimidazole-5-carboxylic acid (1H-indazol-5-yl) -amide
12	2-phenylamino-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		13	2- (pyridin-4-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
14	2- (thiazol-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		15	2-phenylamino-1H-benzimidazole-5-carboxylic acid (1H-benzotriazol-5-yl) -amide
16	2- (pyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1-methyl-1H-indazol-5-yl) -amide
		17	2- (2-chlorophenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

18	2- (4, 5-Dimethylthiazol-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		19	2- (2, 4-dichlorophenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
20	2- (benzothiazol-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		21	2- (4-phenylthiazol-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
22	2- (2-fluorophenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		23	2- (2-ethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
24	2- (2, 4-dichlorophenylamino) -3H-benzimidazole-5-carboxylic acid (1H-benzotriazol-5-yl) -amide
		25	2- (1-isopropyl-1H-imidazol-2-ylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
26	2- (2, 4-dimethylphenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		27	2- (2-isopropylphenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
28	2- (4-chlorophenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		29	2- (naphthalen-1-ylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
30	2- (2-tert-butylphenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		31	2- (biphenyl-2-ylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
32	2- (2-Propylphenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		33	2- (2, 5-dichlorophenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

34	2- (2-methoxyphenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		35	2- (2-trifluoromethylphenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
36	2- (3-methylpyridin-2-ylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		37	2- (2-trifluoromethoxy-phenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
38	2- (3-fluorophenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		39	2- (4-fluorophenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
40	2- (3, 5-difluorophenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		41	2- (2-Butylphenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
42	2- (3-ethyl-6-methyl-pyridin-2-ylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		43	2- (5-chloro-2-methyl-phenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
44	2- (3-fluoro-2-methyl-phenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		45	2- (5-fluoro-2-methyl-phenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
46	2- (3-chloro-2-methyl-phenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		47	2- (1-cyclopentyl-1H-imidazol-2-ylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
48	2- (2-isopropylphenylamino) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

49	2- (2-isopropylphenylamino) -6-morpholin-4-yl-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		50	6- (4-methylpiperazin-1-yl) -2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
51	2- (3, 5-difluorophenylamino) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		52	2- (2, 4-dichlorophenylamino) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
53	6- (4-methylpiperazin-1-yl) -2- (thiazol-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		54	6-morpholin-4-yl-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
55	2- (3, 5-difluorophenylamino) -6-morpholin-4-yl-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		56	2- (2, 4-dichlorophenylamino) -6-morpholin-4-yl-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
57	6-piperidin-1-yl-2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) amide
		58	6- (4-methyl-piperazin-1-yl) -2- (3-methyl-pyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
59	6-morpholin-4-yl-2- (thiazol-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		60	2- (3-methylpyridin-2-ylamino) -6-morpholin-4-yl-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
61	2- (1-isopropyl-1H-imidazol-2-ylamino) -6- (4-methylpiperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		62	2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6- (4-methylpiperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

63	2- (1-isopropyl-1H-imidazol-2-ylamino) -6-morpholin-4-yl-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		64	2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6-morpholin-4-yl-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
65	2- (2-ethyl-2H-pyrazol-3-ylamino) -6-morpholin-4-yl-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		66	2- (2-isopropylphenylamino) -3H-benzimidazole-5-carboxylic acid [3- (2-morpholin-4-yl-ethylamino) -1H-indazol-6-yl]-amides of
67	2- (2-isopropylphenylamino) -3H-benzimidazole-5-carboxylic acid [3- (3-morpholin-4-yl-propylamino) -1H-indazol-6-yl]-amides of
		68	2- (2-isopropylphenylamino) -3H-benzeneBenzimidazole-5-carboxylic acid (3-methylamino-1H-indazol-6-yl) -amide
69	2- (pyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (3-amino-1H-indazol-6-yl) -amide
		70	2- (2-isopropylphenylamino) -3H-benzimidazole-5-carboxylic acid {3- [ (1-methyl-piperidine-4-carbonyl) -amino [ ]]-1H-indazol-6-yl } -amide
71	2- (pyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (3-acetamido-1H-indazol-6-yl) -amide
		72	2- (2, 4-dichlorophenylamino) -3H-benzimidazole-5-carboxylic acid (3-acetamido-1H-indazol-6-yl) -amide
73	2- (2, 4-dichlorophenylamino) -3H-benzimidazole-5-carboxylic acid (3-benzoylamino-1H-indazol-5-yl) -amide
		74	2- (2-isopropylphenylamino) -3H-benzimidazole-5-carboxylic acid (3-methoxy-1H-indazol-6-yl) -amide
75	2- (2-isopropylphenylamino) -3H-benzimidazole-5-carboxylic acid [3- (2-morpholin-4-yl-ethoxy) -1H-indazol-6-yl]-amides of
		76	2- (2, 4-dichlorophenylamino) -3H-benzimidazole-5-carboxylic acid (3-morpholin-4-ylmethyl-1H-indazol-6-yl) -amide

77	2- (2, 4-dichlorophenylamino) -3H-benzimidazole-5-carboxylic acid (3-methyl-1H-indazol-6-yl) -amide
		78	2- (2-ethylphenylamino) -3H-benzimidazole-5-carboxylic acid (3-chloro-1H-indazol-6-yl) -amide
79	2- [6- (1H-indazol-6-ylcarbamoyl) -1H-benzimidazol-2-ylamino]-6, 7-dihydro-4H-thiazolo [5, 4-c]Pyridine-5-carboxylic acid tert-butyl ester
		80	2- (4, 5, 6, 7-tetrahydro-thiazolo [5, 4-c)]Pyridin-2-ylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
81	2- (2-isopropylphenylamino) -3H-benzimidazole-5-carboxylic acid [1- (2-hydroxy-ethyl) -1H-indazol-5-yl]-amides of
		82	2- (2-cyclohexylphenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
83	2- (3-methylthiophen-2-ylamino)) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		84	1H-indazole-6-carboxylic acid [2- (2-isopropyl-phenylamino) -3H-benzimidazol-5-yl]-amides of
85	6- (4-methylpiperazin-1-yl) -2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-5-yl) -amide
		86	6-morpholin-4-yl-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-5-yl) -amide
87	4- [6- (1H-indazol-6-ylcarbamoyl) -2- (2-trifluoromethylphenylamino) -3H-benzimidazol-5-yl]-piperazine-1-carboxylic acid tert-butyl ester
		88	6-piperazin-1-yl-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
89	4- [6- (1H-indazol-6-ylcarbamoyl) -2- (3-methylpyridin-2-ylamino) -3H-benzimidazol-5-yl]-piperazine-1-carboxylic acid tert-butyl ester
		90	2- (3-methylpyridin-2-ylamino) -6-piperazin-1-yl-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

91	2- (2, 6-diethylphenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		92	6-diisobutylamino-2- (2-trifluoromethylphenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
93	6-diethylamino-2- (3-methylpyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		94	6- (2, 6-dimethylmorpholin-4-yl) -2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
95	6-diethylamino-2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		96	6- [ (2-dimethylaminoethyl) methylamino]-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
97	6- (4-Dimethylaminopiperidin-1-yl) -2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		98	6-dipropylamino-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
99	6-dipropylamino-2- (3-methylpyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		100	6- [ bis- (2-methoxyethyl) amino]-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
101	6- (4-hydroxypiperidin-1-yl) -2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		102	6- [ ethyl- (2-methoxyethyl) amino]-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
103	6- [ bis- (2-methoxyethyl) amino]-2- (3-methylpyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		104	2- (3-methylpyridin-2-ylamino) -6-pyrrolidin-1-yl-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

105	6-pyrrolidin-1-yl-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		106	6- [ (2-dimethylaminoethyl) ethylamino]-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
107	6- (4-hydroxypiperidin-1-yl) -2- (3-methylpyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		108	6- [ ethyl- (2-methoxyethyl) amino]-2- (3-methylpyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
109	6- (ethylpropylamino) -2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		110	6- (ethylpropylamino) -2- (3-methylpyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
111	6- (4-isopropylpiperazin-1-yl) -2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		112	6- (Ethylmethylammonium) 6Yl) -2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
113	6- (ethylmethylamino) -2- (3-methylpyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		114	6- (4-isopropylpiperazin-1-yl) -2- (3-methylpyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
115	2- (3-chloro-pyridin-2-ylamino) -6-diethylamino-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		116	6-diethylamino-2- (3-trifluoromethyl-pyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
117	2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6-diethylamino-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		118	2-cyclohexylamino-6-diethylamino-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

119	2-cyclopentylamino-6-diethylamino-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		120	2- (bicyclo [2.2.1]]Hept-2-ylamino) -6-diethylamino-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
121	6-diethylamino-2-isopropylamino-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		122	6-diethylamino-2- (3-ethyl-6-methyl-pyridin-2-ylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
123	6-diethylamino-2- (2, 5-difluoro-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		124	6-diethylamino-2- (3, 5-difluoro-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
125	2- (2-chloro-5-trifluoromethyl-phenylamino) -6-diethylamino-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		126	6-diethylamino-2- (2-trifluoromethoxy-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
127	3- [ 6-diethylamino-5- (1H-indazol-6-ylcarbamoyl) -1H-benzimidazol-2-ylamino]-benzoic acid methyl ester
		128	6-diethylamino-2- (2-isopropyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
129	2- (4-chloro-phenylamino) -6-diethylamino-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		130	2- (2, 4-dichloro-phenylamino) -6-diethylamino-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
131	6-diethylamino-2- (2, 6-difluoro-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		132	6-diethylamino-2- (2-methoxy-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

133	6-diethylamino-2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide
		134	2- (bicyclo [2.2.1]]Hept-2-ylamino) -6-diethylamino-1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide
135	6-diethylamino-2-isopropylamino-1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide
		136	6-diethylamino-2- (2, 5-difluoro-phenylamino) -3H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide
137	6-diethylamino-2- (3, 5-difluoro-phenylamino) -3H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide
		138	2- (2, 4-dichloro-phenylamino) -6-diethylamino-3H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide
139	6-diethylamino-2- (2-trifluoromethoxy-phenylamino) -3H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide
		140	6-diethylamino-2- (2-isopropyl-phenylamino) -3H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide
141	6- (4-methyl-piperazin-1-yl) -2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide
		142	6- (4-methyl-piperazin-1-yl) -2- (3-methyl-pyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide
143	6-morpholin-4-yl-2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide
		144	2- (3-methyl-pyridin-2-ylamino) -6-morpholin-4-yl-1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide
145	6- (3, 5-dimethyl-piperazin-1-yl) -2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		146	6- (2-methoxyethylamino) -2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

147	6- (2-methoxy-ethylamino) -2- (3-methyl-pyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		148	6- (4-methyl-piperazin-1-yl) -2- (2-trifluoromethyl-benzylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
149	2-benzylamino-6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		150	2- (cyclohexylmethyl-amino) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
151	2-cyclopentylamino-6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		152	2- ((1S, 2S, 4R) -bicyclo [2.2.1]Hept-2-ylamino) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
153	2- (adamantan-1-ylamino) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		154	6-propylamino-2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
155	{1- [6- (1H-indazol-6-ylcarbamoyl) -2- (2-trifluoromethyl-phenylamino) -3H-benzimidazol-5-yl]-piperidin-4-yl } -carbamic acid tert-butyl ester
		156	6- (4-Ammonia)1-piperidin-1-yl-2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide trihydrochloride
157	{1- [6- (1H-indazol-6-ylcarbamoyl) -2- (3-methyl-pyridin-2-ylamino) -3H-benzimidazol-5-yl]-piperidin-4-yl } -carbamic acid tert-butyl ester
		158	6- (4-amino-piperidin-1-yl) -2- (3-methyl-pyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide trihydrochloride
159	[5- (1H-indazol-6-ylethynyl) -1H-benzimidazol-2-yl]- (2-trifluoromethyl-phenyl) -amines
		160	6-dimethylamino-2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

161	6-dimethylamino-2- (3-methyl-pyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		162	6- (4-methyl-piperazin-1-yl) -2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid benzothiazol-5-ylamide
163	4- [6- (benzothiazol-6-ylcarbamoyl) -2- (2-trifluoromethyl-phenylamino) -3H-benzimidazol-5-yl]-piperazine-1-carboxylic acid tert-butyl ester
		164	6-piperazin-1-yl-2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide trihydrochloride
165	4- [6- (benzothiazol-6-ylcarbamoyl) -2- (3-methyl-pyridin-2-ylamino) -3H-benzimidazol-5-yl]-piperazine-1-carboxylic acid tert-butyl ester
		166	2- (3-methyl-pyridin-2-ylamino) -6-piperazin-1-yl-1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide trihydrochloride
167	2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide
		168	6-piperazin-1-yl-2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (5-methyl-1H-indol-6-yl) -amide
169	4- [2- ((1S, 2S, 4R) bicyclo [2.2.1]Hept-2-ylamino) -6- (1H-indazol-6-ylcarbamoyl) -3H-benzimidazol-5-yl]-piperazine-1-carboxylic acid tert-butyl ester
		170	2- ((1S, 2S, 4R) bicyclo [2.2.1]Hept-2-ylamino6-piperazin-1-yl-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide trihydrochloride
171	6-chloro-2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		172	2- ((1S, 2S, 4R) -bicyclo [2.2.1]Hept-2-ylamino) -6-chloro-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
173	6- [4- (2-hydroxy-ethyl) -piperazin-1-yl]-2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		174	{4- [6- (1H-indazol-6-ylcarbamoyl) -2- (2-trifluoromethyl-phenylamino) -3H-benzimidazol-5-yl]-piperazin-1-yl } -acetic acid

175	6- (4-dimethylsulfamoyl-piperazin-1-yl) -2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		176	{6- [5- (1H-indazol-6-yl) -1H-imidazol-2-yl]-1H-benzimidazol-2-yl } - (2-trifluoromethyl-phenyl) -amine
177	6- (2-dimethylamino-ethylsulfanyl) -2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		178	5-Ethyl-8- (1H-indazol-6-yl) -2- (2-trifluoromethyl-phenylamino) -5, 6, 7, 8-tetrahydro-3H-1, 3, 5, 8-tetraaza-cyclohepta [ f]Indene-9-ones
179	6-imidazol-1-yl-2- (2-trifluoromethyl-phenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		180	2- (2-trifluoromethyl-phenylamino) -benzooxazole-5-carboxylic acid (1H-indazol-6-yl) -amide
181	2- (1-benzyl-1H-imidazol-2-ylamino) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		182	4- [2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6- (1H-indazol-6-ylcarbamoyl) -3H-benzimidazol-5-yl]-piperazine-1-carboxylic acid tert-butyl ester
183	2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6-piperazin-1-yl-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide trihydrochloride
		184	2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6- (4-isopropyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
185	2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6- (4-ethyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		186	2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6- [ (2-dimethylamino-ethyl) -methyl-amino]-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
187	2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6- (4-methyl- [1, 4 [)]Diazepan-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		188	2- (1-cyclohexyl-1H-imidazol-2-ylamino) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

189	2- (1-methyl-1H-imidazol-2-ylamino) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		190	2- (1-cyclohexylmethyl-1H-imidazol-2-ylamino) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -acylAmines as pesticides
191	2- (1-isobutyl-1H-imidazol-2-ylamino) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		192	2- (1-cyclobutyl-1H-imidazol-2-ylamino) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
193	2- [1- (1-ethyl-propyl) -1H-imidazol-2-ylamino]-6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		194	2- (1-butyl-1H-imidazol-2-ylamino) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
195	2- [1- (2-methoxy-ethyl) -1H-imidazol-2-ylamino]-6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		196	2- (1-ethyl-1H-imidazol-2-ylamino) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
197	2- [1- (2-methoxy-ethyl) -1H-imidazol-2-ylamino]-6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide
		198	2-(1-Ethyl-1H-imidazolylamino) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide
199	2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-5-yl) -amide
		200	2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-benzotriazol-5-yl) -amide
201	2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide
		202	2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (2-oxo-2, 3-dihydro-1H-indol-5-yl) -amide

203	2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indol-6-yl) -amide
		204	2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6- (4-methyl-piperazine-1-yl) -1H-benzimidazole-5-carboxylic acid (3H-benzimidazol-5-yl) -amide
205	2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid benzothiazol-5-ylamide
		206	6- (4-methyl-piperazin-1-yl) -2- (1-thietan-3-yl-1H-imidazol-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
207	2-amino-6- (4-methylpiperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide trihydrobromide salt
		208	2- (3-cyclopentyl-3-ethylureido) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
209	2-mercapto-6- (4-methylpiperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
		210	2- (1-cyclopentyl-1H-benzimidazol-2-ylamino) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide
211	[6- (1H-indazol-6-yloxy) -1H-benzimidazol-2-yl]- (2-trifluoromethyl-phenyl) -amines
		212	{5- [2- (1H-indazol-6-yl) -ethyl]-1H-benzoImidazol-2-yl } - (2-trifluoromethylphenyl) amine
213	3- [ 6-diethylamino-5- (1H-indazol-6-ylcarbamoyl) -1H-benzimidazol-2-ylamino]-benzoic acid
		214	3- [5- (benzothiazol-6-ylcarbamoyl) -6-diethylamino-1H-benzimidazol-2-ylamino]-benzoic acid methyl ester
215	3- [5- (benzothiazol-6-ylcarbamoyl) -6-diethylamino-1H-benzimidazol-2-ylamino]-benzoic acid

In another aspect, the invention comprises a pharmaceutical composition comprising a compound of formula (I) and a pharmaceutically acceptable carrier, excipient, diluent, or mixture thereof. The invention further provides the use of a compound of formula (I) to inhibit Aurora kinase activity and to treat Aurora kinase-mediated disorders.

The term "lower" as used herein denotes groups having between one and six carbons.

The term "alkyl" as used herein means a straight or branched chain hydrocarbon having from one to ten carbon atoms, optionally substituted, and multiple degrees of substitution are also permissible. Examples of "alkyl" as used herein include, but are not limited to, methyl, n-butyl, t-butyl, n-pentyl, isobutyl, and isopropyl, and the like.

The term "alkylene" as used herein denotes a straight or branched chain divalent hydrocarbon radical having from one to ten carbon atoms, optionally substituted, and multiple degrees of substitution are permitted. Examples of "alkylene" as used herein include, but are not limited to, methylene, ethylene, and the like.

The term "alkylidene" as used herein, means a straight or branched chain trivalent hydrocarbon radical having one to ten carbon atoms, optionally substituted, and multiple degrees of substitution are also permissible. Examples of "alkylene" as used herein include, but are not limited to, methine, ethylidene, and the like.

The term "alkenyl" as used herein denotes a hydrocarbon radical having two to ten carbons and at least one carbon-carbon double bond, optionally substituted, and multiple degrees of substitution are also permissible. Examples of "alkenyl" as used herein include, but are not limited to, 3-dimethyl-but-1-enyl, 4-hex-1-enyl, and the like.

The term "alkenylene" as used herein denotes a straight or branched chain divalent hydrocarbon radical having two to ten carbons and one or more carbon-carbon double bonds, optionally substituted, and multiple degrees of substitution are also permissible. Examples of "alkenylene" as used herein include, but are not limited to, ethylene-1, 2-diyl, propylene-1, 3-diyl, methylene-1, 1-diyl, and the like.

The term "alkynyl" as used herein denotes a hydrocarbon radical having two to ten carbons and at least one carbon-carbon triple bond, optionally substituted, and multiple degrees of substitution are also permissible. Examples of "alkynyl" as used herein include, but are not limited to, 4-hex-1-ynyl, 3-dimethyl-but-1-ynyl, and the like.

The term "alkynylene" as used herein denotes a straight or branched chain divalent hydrocarbon radical having two to ten carbons and one or more carbon-carbon triple bonds, optionally substituted, and multiple degrees of substitution are also permissible. Examples of "alkynylene" as used herein include, but are not limited to, acetylene-1, 2-diyl, propyne-1, 3-diyl, and the like.

The terms "haloaliphatic", "haloalkyl", "haloalkenyl" and "haloalkoxy" as used herein, denote an aliphatic, alkyl, alkenyl or alkoxy group, as the case may be, substituted with one or more halogen atoms.

"cycloalkyl" as used herein means a non-aromatic alicyclic hydrocarbon group, optionally having one or more degrees of unsaturation, having three to twelve carbon atoms, optionally substituted, and multiple degrees of substitution are also permissible. Examples of "cycloalkyl" as used herein include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like.

As used herein, "cycloalkylene" refers to a non-aromatic alicyclic divalent hydrocarbon radical having from three to twelve carbon atoms, optionally with one or more degrees of unsaturation, optionally substituted, and multiple degrees of substitution are also permissible. Examples of "cycloalkylene" as used herein include, but are not limited to, cyclopropyl-1, 1-diyl, cyclopropyl-1, 2-diyl, cyclobutyl-1, 2-diyl, cyclopentyl-1, 3-diyl, cyclohexyl-1, 4-diyl, cycloheptyl-1, 4-diyl, cyclooctyl-1, 5-diyl, and the like.

The term "heterocycle" or the term "heterocyclyl" as used herein denotes a non-aromatic three to twelve membered heterocyclic ring, optionally provided with one or more degrees of unsaturation, containing one or more groups selected from S, SO₂Heteroatom substitution of O or N, optionally substituted, and multiple degrees of substitution are also permissible. Such a ring is optionally fused to one to three other "heterocyclic" or cycloalkyl rings. Examples of "heterocyclyl" include, but are not limited to, tetrahydrofuran, 1, 4-dioxane, 1, 3-dioxane, piperidine, pyrrolidine, morpholine, piperazine, and the like.

The term "heterocyclylene" as used herein denotes a divalent radical of a non-aromatic three-to twelve-membered heterocyclic ring, optionally provided with one or more degrees of unsaturation, containing one or more groups selected from S, SO₂Heteroatoms of O or N, optionally substituted, and multiple degrees of substitution are also permissible. Such a ring may optionally be fused to a ring or cycloalkyl ring of one to three phenyl rings or one to three other "heterocyclic rings". Examples of "heterocyclylene" include, but are not limited to, tetrahydrofuran-2, 5-diyl, morpholine-2, 3-diyl, pyran-2, 4-diyl, 1, 4-dioxane-2, 3-diyl, 1, 3-dioxaneCyclohexane-2, 4-diyl, piperidine-1, 4-diyl, pyrrolidine-1, 3-diyl, morpholine-2, 4-diyl, piperazine-1, 4-diyl, and the like.

The term "aryl" as used herein means a phenyl ring or a phenyl ring fused to one to three phenyl rings, optionally substituted, and multiple degrees of substitution are permissible. Examples of aryl groups include, but are not limited to, phenyl, 2-naphthyl, 1-anthracenyl, and the like.

The term "arylene" as used herein means a divalent radical of a phenyl ring or a divalent radical of a phenyl ring system fused to one to three optionally substituted phenyl rings, optionally substituted, and multiple degrees of substitution are also permissible. Examples of "arylene" include, but are not limited to, benzene-1, 4-diyl, naphthalene-1, 8-diyl, and the like.

The term "heteroaryl" as used herein means a five to seven membered aromatic ring or a polycyclic (up to three rings) aromatic ring containing one or more nitrogen, oxygen or sulfur heteroatoms, wherein N-oxides and sulfur monoxide and sulfur dioxide are permissible heteroaromatic substitutions, optionally substituted, and multiple degrees of substitution are permissible. In the case of a polycyclic heteroaryl aromatic ring system, one or more of the rings may contain one or more heteroatoms. Examples of "heteroaryl" as used herein include, but are not limited to, furan, thiophene, pyrrole, imidazole, pyrazole, triazole, tetrazole, thiazole, oxazole, isoxazole, oxadiazole, thiadiazole, isothiazole, pyridine, pyridazine, pyrazine, pyrimidine, quinoline, isoquinoline, quinazoline, benzofuran, benzothiophene, indole, and indazole, and the like.

The term "heteroarylene" as used herein means a five to seven membered aromatic ring divalent radical or a polycyclic (up to three rings) aromatic ring divalent radical containing one or more nitrogen, oxygen or sulfur heteroatoms, wherein the N-oxides and sulfur monoxide and sulfur dioxide are permissible heteroaromatic substitutions, optionally substituted, and degrees of multiple substitution are permissible. For divalent radicals of a polycyclic heteroaryl aromatic ring system, one or more rings may contain one or more heteroatoms. Examples of "heteroarylene" as used herein include, but are not limited to, furan-2, 5-diyl, thiophene-2, 4-diyl, 1, 3, 4-oxadiazole-2, 5-diyl, 1, 3, 4-thiadiazole-2, 5-diyl, 1, 3-thiazole-2, 4-diyl, 1, 3-thiazole-2, 5-diyl, pyridine-2, 4-diyl, pyridine-2, 3-diyl, pyridine-2, 5-diyl, pyrimidine-2, 4-diyl, quinoline-2, 3-diyl, and the like.

The term "fused cycloalkylaryl" as used herein means one or two cycloalkyl groups fused to an aryl group, the aryl and cycloalkyl groups having two atoms in common, wherein the aryl group is the point of substitution. Examples of "fused cycloalkylaryl" as used herein include 5-indenyl, 5, 6, 7, 8-tetrahydro-2-naphthyl,and the like.

The term "fused cycloalkylarylene" as used herein denotes a fused cycloalkylaryl group, wherein the aryl group is divalent. Examples includeAnd the like.

The term "fused arylcycloalkyl" as used herein means one or two aryl groups fused to a cycloalkyl group, the cycloalkyl and aryl groups having two atoms in common, wherein the cycloalkyl group is the point of substitution. Examples of "fused arylcycloalkyl" as used herein include 1-indenyl, 2-indenyl, 9-fluorenyl, 1- (1, 2, 3, 4-tetrahydronaphthyl),And the like.

The term "fused arylcycloalkylene" as used herein denotes a fused arylcycloalkyl group, wherein the cycloalkyl group is divalent. Examples include 9, 1-fluorenylidene,And the like.

The term "fused heterocyclylaryl" as used herein means one or two heterocyclyl groups fused to an aryl group, the aryl and heterocyclyl groups having two atoms in common, wherein the aryl group is the point of substitution. Examples of "fused heterocyclylaryl" as used herein include 3, 4-methylenedioxy-1-phenyl, and,And the like.

The term "fused heterocyclylarylene" as used herein refers to a fused heterocyclylaryl group, wherein the aryl group is divalent. Examples includeAnd the like.

The term "fused arylheterocyclyl" as used herein means one or two aryl groups fused to a heterocyclyl group, the heterocyclyl and aryl groups having two atoms in common, wherein the heterocyclyl group is the point of substitution. Examples of "fused arylheterocyclyl" as used herein include 2- (1, 3-benzodioxolyl)And the like.

The term "fused arylheterocyclylene" as used herein means a fused arylheterocyclyl group wherein the heterocyclyl group is divalent. Examples includeAnd the like.

The term "fused cycloalkylheteroaryl" as used herein means one or two cycloalkyl groups fused to a heteroaryl group, the heteroaryl and cycloalkyl groups having two atoms in common, wherein the heteroaryl group is the point of substitution. Examples of "fused cycloalkylheteroaryl" as used herein include 5-aza-6-indenyl,and the like.

The term "fused cycloalkylheteroarylene" as used herein denotes a fused cycloalkylheteroaryl group, wherein the heteroaryl group is divalent. Examples includeAnd the like.

The term "fused heteroarylcycloalkyl" as used herein means one or two heteroaryl groups fused to a cycloalkyl group, the cycloalkyl and heteroaryl groups having two atoms in common, wherein the cycloalkyl group is the point of substitution. Examples of "fused heteroarylcycloalkyl" as used herein include 5-aza-1-indenyl,and the like.

The term "fused heteroarylcycloalkylene" as used herein denotes a fused heteroarylcycloalkyl group, wherein the cycloalkyl group is divalent. Examples includeAnd the like.

The term "fused heterocyclylheteroaryl" as used herein means one or two heterocyclyl groups fused to a heteroaryl group, the heteroaryl and heterocyclyl groups having two atoms in common, wherein the heteroaryl group is the point of substitution. Examples of "fused heterocyclylheteroaryl" as used herein include 1, 2, 3, 4-tetrahydro-beta-carbolin-8-yl, and,And the like.

The term "fused heterocyclylheteroarylene" as used herein means a fused heterocyclylheteroaryl group, wherein the heteroaryl group is divalent. Examples includeAnd the like.

The term "fused heteroarylheterocyclyl" as used herein means one or two heteroaryl groups fused to a heterocyclyl group, the heterocyclyl and heteroaryl groups having two atoms in common, wherein the heterocyclyl group is the point of substitution. Examples of "fused heteroarylheterocyclyl" as used herein include-5-aza-2, 3-dihydrobenzofuran-2-yl,And the like.

The term "fused heteroarylheterocyclylene" as used herein means a fused heteroarylheterocyclyl group, wherein the heterocyclyl group is divalent. Examples includeAnd the like.

The term "direct valency" as used herein as part of the specification of a structural variable means that the substituents on both sides (before and after) of the variable are directly linked as considered "direct valency". If two or more consecutive variables are each designated as "direct valency", then those substituents which flank (before and after) the two or more consecutive designated "direct valencies" are directly linked.

The term "alkoxy" as used herein denotes a group R_aO-, wherein R_aIs an alkyl group.

The term "alkenyloxy" as used herein denotes the group R_aO-, wherein R_aIs an alkenyl group.

The term "alkynyloxy" as used herein denotes the group R_aO-, wherein R_aIs an alkynyl group.

The term "alkylthio" as used herein denotes the group R_aS-, wherein R_aIs an alkyl group.

The term "alkenylthio" as used herein denotes the group R_aS-, wherein R_aIs an alkenyl group.

The term "alkynylthio" as used herein denotes the group R_aS-, wherein R_aIs an alkynyl group.

The term "alkylsulfinyl" as used herein denotes the group R_aS (O) -, wherein R_aIs an alkyl group.

The term "alkenylsulfinyl" as used herein denotes the group R_aS (O) -, wherein R_aIs an alkenyl group.

The term "alkynylsulfinyl" as used herein denotes the group R_aS (O) -, wherein R_aIs an alkynyl group.

The term "alkylsulfonyl" as used herein denotes the group R_aSO₂-, wherein R_aIs an alkyl group.

The term "alkenylsulfonyl" as used herein denotes the group R_aSO₂-, wherein R_aIs an alkenyl group.

The term "alkynylsulfonyl" as used herein denotes the group R_aSO₂-, wherein R_aIs an alkynyl group.

The term "acyl" as used herein denotes the group R_aC (O) -, wherein R_aIs alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl or heterocyclyl.

The term "aroyl" as used herein denotes the group R_aC (O) -, wherein R_aIs an aryl group.

The term "heteroaroyl" as used herein denotes the group R_aC (O) -, wherein R_aIs a heteroaryl group.

The term "alkoxycarbonyl" as used herein denotes the group R_aOC (O) -, wherein R_aIs an alkyl group.

The term "acyloxy", as used herein, denotes a group R_aC (O) O-, wherein R_aIs alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl or heterocyclyl.

The term "aroyloxy" as used herein denotes the group R_aC (O) O-, wherein R_aIs an aryl group.

The term "heteroaroyloxy" as used herein denotes a group R_aC (O) O-, wherein R_aIs a heteroaryl group.

The term "optional" as used herein means that the subsequently described event may or may not occur, including both occurring and non-occurring events.

The term "substituted" as used herein means that one or more hydrogens of the indicated moiety is replaced with the indicated substituent, multiple degrees of substitution are permissible unless otherwise specified, so long as the substitution results in a stable or chemically feasible compound. A stable compound or chemically feasible compound is one in which the chemical structure is not substantially altered or maintained intact for a sufficient period of time in the absence of moisture or other chemically reactive conditions at temperatures of about-80 ℃ to about +40 ℃ for at least one week, so as to be useful for therapeutic or prophylactic administration to a patient. The phrase "one or more substituents" as used herein means the number of substituents, equal to one to the maximum possible number of substituents, based on the number of available bonding sites, provided that the conditions of stability and chemical feasibility described above are met.

The term "containing" as used herein may mean being substituted at any position along the alkyl, alkenyl, alkynyl or cycloalkyl substituents as defined above by one or more of any O, S, SO₂In-line substitution of N or N-alkyl, e.g. -CH₂-O-CH₂-、-CH₂-SO₂-CH₂-、-CH₂-NH-CH₃And so on.

Whenever the terms "alkyl" or "aryl" or their prefix roots appear in the name of a substituent (e.g., arylalkoxyaryloxy), they should be construed to include those limitations as set forth above for "alkyl" and "aryl". Indicated carbon atomNumber (e.g. C)_1-10) Shall independently denote the number of carbon atoms of an alkyl, alkenyl, alkynyl or cyclic alkyl moiety or of an alkyl moiety of a larger substituent in which the term "alkyl" appears as its prefix root.

The term "oxo" as used herein shall denote a substituent group ═ O.

The term "halogen" or "halo" as used herein means iodine, bromine, chlorine or fluorine.

The term "mercapto" as used herein denotes the substituent-SH.

The term "carboxy" as used herein denotes the substituent-COOH.

The term "cyano" as used herein denotes the substituent-CN.

The term "aminosulfonyl" as used herein denotes the substituent-SO₂NH₂。

The term "carbamoyl" as used herein denotes the substituent-C (O) NH₂。

The term "thio" as used herein denotes the substituent-S-.

The term "sulfinyl", as used herein, denotes the substituent-S (O) -.

The term "sulfonyl", as used herein, denotes the substituent-S (O)₂-。

These compounds can be prepared according to the following reaction schemes (wherein the variables are as defined hereinbefore or therein) using readily available starting materials and reagents. In these reactions, it is also possible to use variations which are known per se to the person skilled in the art, but which are not mentioned in more detail.

The present invention also provides a method for synthesizing a compound useful as an intermediate for the preparation of the compound of formula (I) and a method for preparing the compound of formula (I). Unless otherwise specified, each structural variable is as defined for formula (I).

As shown in scheme I, diaminobenzoate (1) is reacted with isothiocyanate (6) by heating in a solvent such as, but not limited to THF, to afford thiourea (2). Isothiocyanate (6) is either commercially available or prepared from the corresponding amine (5) and reacted with 1, 1' -thiocarbonylimidazole in a solvent such as, but not limited to, THF. Thiourea (2) is treated with a coupling reagent such as, but not limited to, EDC to form aminobenzimidazole, which is hydrolyzed to give carboxylic acid (3). Carboxylic acid (3) is then coupled with an amine in the presence of a coupling reagent, such as but not limited to HBTU, to form amide (4).

Scheme I

Alternatively, aminobenzimidazole (4) was also prepared as shown in scheme II. Carboxylic acid (7) is coupled with an amine in the presence of a coupling reagent, such as but not limited to HBTU, to form amide (8). The nitro group of intermediate (8) is then reduced under conditions such as, but not limited to, Pd/C under a hydrogen atmosphere to afford diamine (9). Diamine (9) is then reacted with an isothiocyanate as described in scheme I to provide thiourea, which upon treatment with a coupling reagent, such as but not limited to EDC, provides aminobenzimidazole (4).

Scheme II

As shown in scheme III, benzoyl chloride derivatives (10) are obtained from the corresponding carboxylic acids by heating with a reagent such as, but not limited to, oxalyl chloride, and coupling with an amine in the presence of a base such as, but not limited to, pyridine to form amides (11). Amide (11) is then converted to nitroaniline (12) by heating with ammonium hydroxide in a solvent such as, but not limited to, DCM. Nitroaniline (12) is converted to intermediate (13) under neat conditions or in the presence of a solvent, after heating with a nucleophile, such as but not limited to an amine. The nitro group of intermediate (13) is then reduced under conditions such as, but not limited to, Pd/C under a hydrogen atmosphere to provide diamine (14). Diamine (14) is then reacted with an isothiocyanate as described in scheme I to provide a thiourea, which upon treatment with a coupling reagent, such as but not limited to EDC, provides aminobenzimidazole (15).

Scheme III

The compounds of the invention are inhibitors of Aurora kinase. The ability of a compound to inhibit Aurora kinase can be determined in vitro. In vitro assays include assays that determine the ability to inhibit phosphorylation of a substrate protein or peptide by an Aurora kinase. Alternatively, in vitro assays quantify the ability of a compound to bind to Aurora kinase. The binding of the inhibitor can be measured by radiolabelling the inhibitor prior to binding, isolating the inhibitor/Aurora kinase complex and determining the amount of radiolabel bound. Alternatively, the binding of the inhibitor can be determined by performing a competition experiment in which the new inhibitor and Aurora kinase bound to a known radioligand are incubated. The ability of a compound to affect a cellular or physiological function mediated by Aurora kinase activity can also be determined. Assays for each of these activities are described in the examples and/or are known in the art.

In general, embodiments of the invention useful for pharmaceutical applications may have an inhibitory potency (IC) of less than about 10 μ M for the protein of interest₅₀). In one embodiment, an embodiment of the invention useful for pharmaceutical applications may have an IC of less than about 1 μ M for the protein of interest₅₀. For certain applications, lower inhibitory potency may be useful. Thus, in another embodiment, the compounds of the invention inhibit the IC of Aurora kinase₅₀May be in the range of less than 100 nM. In another kindIn an embodiment, the inhibitory potency (IC) of the compounds of the invention to inhibit Aurora kinase₅₀) Can be between 0.1nM and 100 nM.

In another embodiment, the present invention provides a pharmaceutical composition comprising a compound of formula (I), wherein the compound of formula (I) is administered in a dose of less than 1,000mg/kg body weight per day. In another embodiment, the present invention provides a pharmaceutical composition comprising a compound of formula (I), wherein the compound of formula (I) is administered at a dose of less than 100mg/kg body weight per day. In another embodiment, the present invention provides a pharmaceutical composition comprising a compound of formula (I), wherein the compound of formula (I) is administered at a dose of less than 10mg/kg body weight per day.

Embodiments of the compounds of the present invention demonstrate utility as inhibitors of Aurora kinase activity or as inhibitors of cell proliferation. Embodiments of the invention described herein are additionally directed to pharmaceutical compositions and methods of inhibiting Aurora kinase in a subject, these methods comprising administering to a subject in which inhibition of Aurora kinase activity is desired a therapeutically effective amount of a compound of formula (I) as defined above, as a single or polymorphic crystalline form, amorphous form, single enantiomer, racemic mixture, single stereoisomer, mixture of stereoisomers, single diastereomer, mixture of diastereomers, solvate, pharmaceutically acceptable salt, solvate, prodrug, biohydrolyzable ester, or biohydrolyzable amide thereof.

In one embodiment, the present invention provides a method of inhibiting Aurora kinase activity comprising contacting a cell in which inhibition of Aurora kinase is desired with an Aurora kinase inhibitor of formula (I). In one embodiment, the Aurora kinase inhibitor acts on and reduces a small fraction of Aurora kinase activity in the cell. If a compound of the invention selectively acts as an Aurora kinase inhibitor in preference to one or more other kinases, treatment of a subject with such a selective compound may provide an advantage over non-specific kinase inhibitors in the treatment of cancer in a subject. Thus, in another embodiment, the invention provides a method of selectively inhibiting Aurora kinase activity in the presence of one or more other kinases, comprising contacting a cell in which inhibition of Aurora kinase is desired with an Aurora kinase inhibitor of formula (I).

The method according to this aspect results in inhibition of cell proliferation of the contacted cells. The expression "inhibiting cell proliferation" is used to indicate the ability of an Aurora kinase inhibitor to inhibit the number of cells or cell growth in contacted cells compared to cells not contacted with the inhibitor. The assessment of cell proliferation can be performed by counting cells using a cytometer, measuring uptake of labeled nucleotides or nucleotide analogs, or determining cell viability. If the cell is in solid growth (e.g., a solid tumor or organ), such an assessment of cell proliferation can be made by measuring the growth, e.g., with a caliper, and comparing the size of the growth of the contacted cells to the size of the growth of the non-contacted cells.

The growth of cells contacted with the inhibitor may be delayed by at least about 50% compared to the growth of non-contacted cells. In various embodiments, cell proliferation of the contacted cells is inhibited by at least about 75% compared to non-contacted cells. In some embodiments, the phrase "inhibiting cell proliferation" includes a decrease in the number of contacted cells as compared to non-contacted cells. Thus, an Aurora kinase inhibitor that inhibits cell proliferation of a contacted cell may induce the contacted cell to undergo growth retardation, to undergo growth arrest, to undergo apoptosis (i.e., apoptosis), or to undergo necrotic cell death.

Subjects may include, but are not limited to, horses, cattle, sheep, pigs, mice, dogs, cats, primates, such as chimpanzees, gorillas, rhesus monkeys, and humans. In one embodiment, the subject is a human in need of inhibition of Aurora kinase activity.

Pharmaceutical compositions containing the compounds of the present invention 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, syrups or elixirs. Compositions for oral use may be prepared according to any known method and such compositions may contain one or more agents selected from the group consisting of sweetening agents, flavouring agents, colouring agents and preserving agents in order to provide pharmaceutically elegant and palatable preparations. Tablets may 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, such as corn starch or alginic acid; binding agents, such as starch, gelatin or acacia; and lubricating agents, such as magnesium stearate, stearic acid or talc. 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. The osmotic therapeutic tablets for controlled release can also be prepared using prior art coatings.

The oral formulations 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 soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, such as peanut oil, liquid paraffin, or olive oil.

Aqueous suspensions may contain the active compound in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, for example sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, 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 heptadecenoxycetyl alcohol, 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 polyoxyethylene sorbitan monooleate. Aqueous suspensions may also contain 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. 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. Antioxidants, such as ascorbic acid, may be added to these compositions for preservation.

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

The pharmaceutical composition of the present invention may also be in the form of an oil-in-water emulsion. 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 thereof. 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 sterile injectable aqueous or oleaginous suspensions. Such suspensions may be formulated according to known methods using suitable dispersing or wetting agents and suspending agents as described 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-butanediol. Among the acceptable carriers and solvents that may be employed are water, sterile water for injection (SWFI), sexagesimal solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any brand of fixed oil may be used, using synthetic mono-or diglycerides. In addition, fatty acids, such as oleic acid, are used in the preparation of injectables.

Thus, in another embodiment, the present invention provides a pharmaceutical formulation solution comprising a compound of formula (I) or a salt thereof.

The solutions of the invention can be provided in sealed containers, especially made of glass, in unit-dose form or in multi-dose form.

Any pharmaceutically acceptable salt of the compound of formula (I) may be used to prepare the solutions of the present invention. Examples of suitable salts may be, for example, salts of mineral inorganic acids, such as hydrochloride, hydrobromide, sulphate, phosphate, nitrate and the like, and salts of certain organic acids, such as acetate, succinate, tartrate, ascorbate, citrate, glutamate, benzoate, methanesulphonate, ethanesulphonate and the like. In one embodiment, the compound of formula (I) is a hydrochloride salt, including mono-, di-or tri-hydrochloride salts.

Any pharmaceutically acceptable solvent which is capable of dissolving the compound of formula (I) or a pharmaceutically acceptable salt thereof may be used. The solutions of the present invention may also contain one or more additional components, such as co-solvents (which may be the same as the solvent), tonicity adjusting agents, stabilizers, preservatives, or mixtures thereof. Examples of solvents, co-solvents, tonicity adjusting agents, stabilizers and preservatives that may be suitable for solution formulations are described below.

Suitable solvents and co-solvents may include, but are not limited to, water; sterile water for injection (SWFI)(ii) a Physiological saline; alcohols such as ethanol, benzyl alcohol, etc.; glycols and polyols such as propylene glycol, glycerol, and the like; esters of polyhydric alcohols such as diacetin, triacetin, and the like; polyglycols and polyethers such as polyethylene glycol 400, propylene glycol methyl ether, and the like; dioxolanes such as isopropylidene glycerol and the like; dimethyl isosorbide; pyrrolidone derivatives such as 2-pyrrolidone, N-methyl-2-pyrrolidone, polyvinylpyrrolidone (a cosolvent only), and the like; polyoxyethylated fatty alcohols; esters of polyoxyethylenated fatty acids; polysorbates, e.g. Tween^TM(ii) a Polyoxyethylene derivatives of polypropylene glycols, e.g. Pluronics^TM。

Suitable tonicity adjusting agents may include, but are not limited to, pharmaceutically acceptable inorganic chlorides such as sodium chloride; glucose; lactose; mannitol; sorbitol, and the like.

Preservatives suitable for physiological administration may be, for example, esters of p-hydroxybenzoic acid (e.g. methyl, ethyl, propyl and butyl esters or mixtures thereof), chlorocresol and the like.

Suitable stabilizers include, but are not limited to, monosaccharides (e.g., galactose, fructose, and dunaliose), disaccharides (e.g., lactose), polysaccharides (e.g., dextran), cyclic oligosaccharides (e.g., alpha-, beta-, gamma-cyclodextrins), aliphatic polyols (e.g., mannitol, sorbitol, and thioglycerol), cyclic polyols (e.g., inositol), and organic solvents (e.g., ethanol and glycerol).

The above-mentioned solvents and co-solvents, tonicity adjusting agents, stabilizers and preservatives may be used alone or as a mixture of two or more in a solution formulation.

In one embodiment, the pharmaceutical solution formulation may comprise a compound of formula (I) or a pharmaceutically acceptable salt thereof, SWFI, and an agent selected from the group consisting of: sodium chloride solution (i.e. physiological saline), glucose, mannitol or sorbitol, wherein the content of the agent is less than or equal to 5%. The pH of such a formulation may also be adjusted using pharmaceutically acceptable acids or bases to improve storage stability.

In the solutions of the invention, the concentration of the compound of formula (I) or a pharmaceutically acceptable salt thereof may be less than 100mg/mL, or less than 50mg/mL, or less than 10mg/mL and greater than 0.01mg/mL, or between 0.5mg/mL and 5mg/mL, or between 1mg/mL and 3 mg/mL.

Suitable packaging for the pharmaceutical solution preparation can be all containers approved for parenteral use, such as plastic and glass containers, ready-to-use syringes, and the like. In one embodiment, the container is a sealed glass container, such as a vial or ampoule. Fusion-sealed glass vials are particularly preferred.

According to an embodiment of the invention, a sterile injectable solution comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof in a physiologically acceptable solvent having a pH of 2.5 to 3.5 is provided in a sealed glass container. In the case of solution formulations, various compounds of the present invention may remain dissolved or stable for extended periods of time in solutions having a pH below 6. Furthermore, the acid salts of the compounds of the present invention may be more soluble in aqueous solutions than their free base counter-moieties, but upon addition of the acid salt to an aqueous solution, the pH of the solution may be too low to be suitable for administration. Thus, a solution formulation having a pH of 4.5 or greater may be combined with a diluent solution having a pH greater than 7 prior to administration such that the pH of the combined formulation administered is pH 4.5 or greater. In one embodiment, the diluent solution comprises a pharmaceutically acceptable base, such as sodium hydroxide. In another embodiment, the pH of the diluent solution is between 10 and 12. In another embodiment, the pH of the administered combination formulation is greater than 5.0. In another embodiment, the pH of the combined preparation administered is between pH 5.0 and 7.0.

The invention also provides a process for the manufacture of a sterile solution having a pH of from 2.5 to 3.5, which process comprises dissolving a compound of formula (I), or a pharmaceutically acceptable salt thereof, in a pharmaceutically acceptable solvent. If a pharmaceutically acceptable acid salt of a compound of formula (I) is used, the pH of the solution may be adjusted using a pharmaceutically acceptable base or alkaline solution, and a physiologically acceptable acid or buffer may be added to adjust the pH to within the desired range. The method may further comprise passing the resulting solution through a sterilizing filter.

One or more additional components, such as co-solvents, tonicity adjusting agents, stabilizers, and preservatives, such as those previously specified, may be added to the solution prior to passing the solution through the sterile filter.

Specific drug solution formulations having different pH and concentration are illustrated in the following examples.

Thus, according to the present invention, there is also provided a method of inhibiting the growth of a tumor or cancer, comprising administering to a host suffering from said tumor or cancer an injectable solution according to the present invention containing an active drug substance in an amount sufficient to inhibit the growth of said tumor.

The injectable solutions of the present invention may be administered by rapid intravenous injection or infusion in accordance with a variety of possible dosage regimes.

The compositions may also be in the form of suppositories for rectal administration of the compounds of the invention. 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 include, for example, cocoa butter and polyethylene glycols.

For topical use, creams, ointments, jellies, solutions or suspensions, etc., containing the compounds of the present invention are contemplated. For the purposes of this application, topical application should include mouth washes and mouth washes.

The compounds of the invention may also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be made from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines.

The invention also provides prodrugs of the invention. Pharmaceutically acceptable salts of the compounds of the present invention are also included within the scope of the present invention if basic or acidic groups are present in the structure. The term "pharmaceutically acceptable salts" refers to non-toxic salts of the compounds of the present invention, which are generally prepared by reacting the free base with a suitable organic or inorganic acid, or by reacting an acid with a suitable organic or inorganic base. Representative salts include the following: acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, calcium ethylenediaminetetraacetate, camphorsulfonate, carbonate, chloride, clavulanate, citrate, dihydrochloride, ethylenediaminetetraacetate, edisylate, estolate, ethanesulfonate, fumarate, glucoheptanoate, gluconate, glutamate, hydroxyacetyl-p-aminobenzoate, hexylresorcinol, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isethionate, lactate, lactobionate, laurate, malate, maleate, mandelate, methanesulfonate, methyl bromide, methyl nitrate, methyl sulfate, monopotassium maleate, mucate, naphthalenesulfonate, nitrate, N-methylglucamine, oxalate, pamoate, palmitate, calcium ethylenediaminetetraacetate, camphorate, napsylate, trinitrate, N-methylglucamine, oxalate, pamoate, napsylate, and mixtures thereof, Pantothenate, phosphate/diphosphate, polygalacturonate, potassium, salicylate, sodium, stearate, subacetate, succinate, tannate, tartrate, teoclate, tosylate, triethyliodide, trimethylammonium, and valerate. When an acidic substituent, for example, -COOH, is present, salts of ammonium, morpholinium, sodium, potassium, barium, calcium, and the like can be formed and used as a dosage form. When a basic group, such as an amino group or a basic heteroaryl radical, such as pyridyl, is present, an acidic salt, such as hydrochloride, hydrobromide, phosphate, sulfate, trifluoroacetate, trichloroacetate, acetate, oxalate, maleate, pyruvate, malonate, succinate, citrate, tartrate, fumarate, mandelate, benzoate, cinnamate, methanesulfonate, ethanesulfonate, picrate, etc., including acids related to the pharmaceutically acceptable salts listed in Journal of Pharmaceutical Science, 66, 2(1977) p.1-19, can be formed.

Other salts which are not pharmaceutically acceptable may be used in the preparation of the compounds of the invention and they constitute further inventive aspects.

In addition, some of the compounds of the present invention may form solvates with water or common organic solvents. Such solvates are also encompassed within the scope of the present invention.

Thus, in a further embodiment, there is provided a pharmaceutical composition comprising a compound of the present invention, or a pharmaceutically acceptable salt, solvate or prodrug thereof, and a pharmaceutically acceptable carrier, excipient, diluent or mixture thereof.

The pharmaceutical compositions of the invention may be used in therapeutic applications involving Aurora kinase-mediated conditions. The term "Aurora kinase-mediated condition" as used herein includes any condition, disease or disorder which is caused by or characterized by either increased expression or activity of Aurora kinase, or which requires Aurora kinase activity. The term "Aurora kinase-mediated condition" also includes any condition, disease or disorder in which inhibition of Aurora kinase activity is beneficial. Aurora kinase-mediated conditions include proliferative disorders. Non-limiting examples of proliferative disorders include chronic inflammatory proliferative disorders, such as psoriasis and rheumatoid arthritis and chronic lung disease; proliferative ocular disorders, such as diabetic retinopathy; benign proliferative disorders, such as hemangiomas; restenosis, atherosclerosis, angiogenesis and cancer.

In one embodiment, the composition is formulated for administration to a subject suffering from or at risk of developing or experiencing a relapse of an Aurora kinase-mediated disorder. In one embodiment, the pharmaceutical compositions of the present invention are those formulated for oral, intravenous, or subcutaneous administration. However, any such dosage form containing a therapeutically effective amount of a compound of the present invention is within the purview of routine experimentation and is therefore within the scope of the present invention. In some embodiments, the pharmaceutical compositions of the present invention may further comprise another therapeutic agent. In one embodiment, such other therapeutic agents are those that are normally administered to a subject suffering from the disease or condition being treated.

As used herein, a "therapeutically effective amount" is an amount of a compound of formula (I) sufficient to cause a detectable reduction in Aurora kinase activity or the severity of an Aurora kinase-mediated condition. The amount of Aurora kinase inhibitor required will depend on the effectiveness of the inhibitor for a given cell type and the length of time required to treat the condition. It will also be understood that the specific dose and regimen of treatment for any particular subject will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex and diet of the patient, the time of administration, the rate of excretion, drug combination, the judgment of the attending physician, and the severity of the particular disease being treated.

In another aspect, the invention provides a method of treating a subject suffering from or at risk of developing or experiencing a recurrence of an Aurora kinase-mediated disorder. The method comprises the step of administering to the subject a compound or pharmaceutical composition according to the invention. The compounds and pharmaceutical compositions of the invention may be used to achieve a beneficial therapeutic or prophylactic effect, for example in subjects suffering from a proliferative disorder as discussed above, such as cancer.

The term "cancer" as used herein means a cellular disorder characterized by uncontrolled or unregulated cell proliferation, reduced cell differentiation, inappropriate ability to invade surrounding tissues, and/or the ability to establish new growth at ectopic sites. The term "cancer" includes, but is not limited to, solid tumors and hematological derived tumors. The term "cancer" encompasses diseases of the skin, tissue, organs, bone, cartilage, blood and blood vessels. The term "cancer" further encompasses primary and metastatic cancers.

Non-limiting examples of solid tumors that can be treated with the methods of the invention include pancreatic cancer; bladder cancer; colorectal cancer; breast cancer, including metastatic breast cancer; prostate cancer, including androgen-dependent and androgen-independent prostate cancer; kidney cancers, including, for example, metastatic renal cell carcinoma; hepatocellular carcinoma; lung cancer, including, for example, non-small cell lung cancer (NSCLC), bronchoalveolar carcinoma (BAC), and lung adenocarcinoma; ovarian cancer, for example, including progressive epithelial or primary peritoneal cancer; cervical cancer; gastric cancer; esophageal cancer; head and neck cancer, e.g., squamous cell carcinoma including head and neck; melanoma; neuroendocrine cancers, including metastatic neuroendocrine tumors; brain tumors, including, for example, glioma, degenerative oligodendroglioma, adult glioblastoma multiforme, and adult degenerative astrocytoma; bone cancer; and soft tissue sarcomas.

In some other embodiments, the cancer is a hematologic malignancy. Non-limiting examples of hematological malignancies include Acute Myeloid Leukemia (AML); chronic Myelogenous Leukemia (CML), including accelerated CML and CML blast phase (CML-BP); acute Lymphoblastic Leukemia (ALL); chronic Lymphocytic Leukemia (CLL); hodgkin's Disease (HD); non-hodgkin's lymphoma (NHL) including follicular lymphoma and adventitial cell lymphoma; b-cell lymphoma; t-cell lymphoma; multiple Myeloma (MN); waldenstrom's macroglobulinemia; myelodysplastic syndromes (MDS) including Refractory Anemia (RA), refractory anemia with ringed sideblast (RARS), refractory anemia with excess blasts (RAEB) and transformed RAEB (RAEB-T); and myeloproliferative syndromes.

In some embodiments, the compounds or compositions of the invention are used to treat cancer in which Aurora kinase activity is amplified. In some embodiments, the compounds or compositions of the invention are used to treat a patient having or at risk of developing or experiencing a recurrence of a cancer selected from the group consisting of: colorectal cancer, ovarian cancer, breast cancer, gastric cancer, prostate cancer, and pancreatic cancer. In certain embodiments, the cancer is selected from the group consisting of: breast cancer, colorectal cancer, and pancreatic cancer.

In some embodiments, the Aurora kinase inhibitors of the invention are administered in combination with another therapeutic agent. Other therapeutic agents may also inhibit Aurora kinase or may act by a different mechanism. In some embodiments, the additional therapeutic agent is one that is normally administered to a subject suffering from the disease or condition being treated. The Aurora kinase inhibitors of the present invention may be administered in a single dosage form with other therapeutic agents or as separate dosage forms. When administered as a separate dosage form, the other therapeutic agent may be administered prior to, simultaneously with, or subsequent to the administration of the Aurora kinase inhibitor of the invention.

In some embodiments, the Aurora kinase inhibitors of the invention are administered in combination with a therapeutic agent selected from the group consisting of: cytotoxic agents, radiation therapy, immunotherapy or other kinase inhibitors. Non-limiting examples of cytotoxic agents that may be suitable for use in combination with the Aurora kinase inhibitors of the present invention include: antimetabolites including, for example, capecitabine (capecitabine), gemcitabine, 5-fluorouracil or 5-fluorouracil/folinic acid, fludarabine, cytarabine, mercaptopurine, thioguanine, pentostatin, and methotrexate; topoisomerase inhibitors, including, for example, etoposide, teniposide, camptothecin, topotecan, irinotecan, doxorubicin, and daunorubicin; vinca alkaloids, including, for example, vincristine and vinblastine; taxanes, including, for example, paclitaxel and docetaxel; platinum agents, including, for example, cisplatin, carboplatin, and oxaliplatin; antibiotics, including, for example, actinomycin D, bleomycin, mitomycin C, doxorubicin (adriamycin), daunorubicin, idarubicin, doxorubicin (doxorubicin), and pegylated liposomal doxorubicin; alkylating agents, such as melphalan, chlorambucil, busulfan, thiotepa, ifosfamide, carmustine, lomustine, semustine, streptozocin, decarbazine, and cyclophosphamide; thalidomide; protein tyrosine kinase inhibitors, including, for example, imatinib mesylate, erlotinib, and gefitinib; antibiotics, including, for example, trastuzumab, rituximab, cetuximab, and bevacizumab; mitoxantrone; dexamethasone; prednisone; and temozolomide.

Examples

The invention may be further understood by reference to the following non-limiting examples. Examples of compounds of the invention and processes that may be used to prepare and identify useful compounds of the invention are described below. Abbreviations used in the examples are as follows:

boc ═ tert-butoxycarbonyl

DCE ═ 1, 2-dichloroethane

DCM ═ dichloromethane

DIEA is diisopropylethylamine

DMF ═ N, N-dimethylformamide

EDC ═ 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride

EtOAc ═ ethyl acetate

HBTU ═ O-benzotriazol-1-yl-N, N' -tetramethyluronium hexafluorophosphate

N-methylpyrrolidine and NMP

THF ═ tetrahydrofuran

In parallel MUX^TMLC-MS data were obtained on the system using gradient elution and four Waters 1525 binary HPLC pumps were run equipped with Mux-UV 2488 multichannel UV-Vis detectors (recorded at 215 and 254 nM) and Leap Technologies HTS PAL autosampler using Waters Xterra MS C184.6X 50mm columns. A three minute gradient was run from 25% B (97.5% acetonitrile, 2.5% water, 0.05% TFA) and 75% a (97.5% water, 2.5% acetonitrile, 0.05% TFA) to 100% B. The system interface is a Waters Micromass ZQ mass spectrometer and electrospray ionization is adopted. All MS data is obtained in positive mode (positive mode) unless otherwise noted. 1H NMR data were obtained on a Varian 400MHz spectrometer.

General procedure a: preparation of isothiocyanates

1, 1' -Thiocarbonylimidazole (1.1mmol) was added to a solution of the amine (1mmol) in THF/DMF (2mL, 1: 1) and the reaction mixture was stirred at 65-70 ℃ for 1 h. The product thus formed is used without isolation for further conversion.

General procedure B: thiourea formation and conversion thereof to aminobenzimidazole

To a solution of isothiocyanate (1mmol) in THF/DMF (2mL, 1: 1) at room temperature was added phenylenediamine (1mmol) and the contents were stirred at room temperature for 2 h. EDC (1.2mmol) was then added to the reaction mixture and the contents were stirred at 65-70 ℃ for 1 h. The reaction mixture was then cooled to room temperature, poured into ice-cold water (10mL), and the solid collected by filtration. The crude product was purified by flash column chromatography using DCM/methanol as eluent.

General procedure C: hydrolysis of benzoic acid esters

A solution of LiOH (12mmol) in water (5mL) was added to a solution of the ester (3mmol) in 1: 1THF/MeOH (10mL) and the resulting mixture was stirred at reflux for 6 h. The reaction mixture was cooled to room temperature and the organic solvent was removed in vacuo. The pH of the resulting suspension was adjusted to pH 6 by dropping 10% aq. HCl, the resulting precipitate was collected by filtration, washed with water and dried under vacuum. The desired carboxylic acid obtained was used without further purification.

General operation D: amide formation using coupling agents

HBTU (1.2mmol) was added in one portion to a solution of carboxylic acid (1.0mmol) in anhydrous DMF or NMP (2.5mL) and the reaction mixture was stirred at room temperature for-30 min. Then amine (1.1mmol) and DIEA (1.5mmol) were added to the reaction mixture and the resulting mixture was stirred at room temperature for 6-12h or at 70-80 ℃ for 1-3 h. The contents were diluted with ice cold water (20mL) and the product precipitated out. After filtration, the pure product is isolated by washing with water and ethyl acetate or by column chromatography on silica gel using DCM/methanol as eluent.

General operation E: formation of amides from acid chlorides

Oxalyl chloride (10mmol) was added to a suspension of carboxylic acid (2mmol) in anhydrous DMF (10 μ L) in anhydrous DCM (4mL) and the mixture was stirred at 50 ℃ for 6-12 h. The mixture was cooled to room temperature and the solvent was removed in vacuo to afford the acid chloride. Toluene (5mL) was added to the acid chloride and the solvent was removed in vacuo to dryness. This procedure was repeated to ensure complete removal of residual oxalyl chloride. The resulting acid chloride was dissolved in anhydrous DCM (2mL) and added dropwise to a suspension of pyridine (0.5mL) in amine (2mmol) in anhydrous DCM (5mL) at 0 deg.C. The mixture was allowed to warm to room temperature and stirred for 3-5 h. The organic volatiles were removed in vacuo and the resulting precipitate was suspended in water (20mL) and collected by filtration followed by water washing (20 mL). The resulting amide was used without further purification.

General operation F: reduction of nitro groups to amines

10% Pd/C (0.1g) was added to a solution of the nitro compound (10mmol) in THF/MeOH (1: 1, 50 mL). The resulting mixture was cooled to room temperature and H₂Stirring for 12h under the atmosphere. The contents were then filtered through a pad of celite and the solid was washed with several portions of methanol. The filtrate and washings were combined and evaporated to give the corresponding amine which was used in the next step without further purification.

General operation G: ortho-nitro-halogenated aromatics substituted with the amino home position (Ipso)

To a suspension of o-nitrohaloarene (10mmol) in methanol (40mL) was added concentrated NH₄Aqueous OH (10 mL). The mixture was heated at 50-60 ℃ for 4 h. The reaction mixture was concentrated in vacuo and the resulting precipitate was collected by filtration, washed with water (50mL) and dried under vacuum to give the corresponding o-nitroaniline which was used for further transformation without further purification.

General procedure H: para-nitro-haloarenes substituted in situ with amines

The mixture of p-nitro haloarene (5mmol) and amine (excess) was heated at 90 ℃ either neat or in dioxane for 1-3 h. The volatiles were removed in vacuo and the resulting residue was suspended in ice-cold water (50mL) with stirring. The resulting precipitate was collected by filtration, washed with water and dried under vacuum to give the desired product, which was used for further transformation without further purification.

Example 1

Synthesis of 2- (isoquinolin-3-ylamino) -1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide

3-Isothiocyanoisoquinoline was prepared from 3-aminoisoquinoline (5mmol) as described in general procedure A.

Reaction of the isothiocyanate with methyl 3, 4-diaminobenzoate (5mmol) followed by cyclization using EDC gave methyl 2- (isoquinolin-3-ylamino) -1H-benzimidazole-5-carboxylate as described in general procedure B. The ester is hydrolyzed using general procedure C to give the corresponding carboxylic acid.

Benzothiazol-6-ylamine (0.25mmol) was coupled with the above carboxylic acid using HBTU using general procedure D to give 2- (isoquinolin-3-ylamino) -1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide. MS: m/z 437(M + H)⁺。

The following compounds, as shown in table 2, were synthesized using the procedure described in example 1.

TABLE 2

Examples	Ar	R	MS(m/z)
				2	Isoquinolin-3-yl	1H-indazol-5-yl	420
3	Pyridin-2-yl	1H-indazol-5-yl	370
				4	Pyridin-2-yl	2-methyl-benzoxazol-5-yl	385
5	Pyridin-2-yl	1H-indazol-6-yl	370

6	Pyridin-3-yl	1H-indazol-6-yl	370
				7	Pyridin-2-yl	Benzothiazol-6-yl	387
8	Pyridin-2-yl	1H-benzotriazol-5-yl	371
				9	2, 4-dichlorophenyl	1-methyl-1H-indazol-5-yl	452
10	2, 4-dichlorophenyl	1H-indazol-5-yl	437
				11	Phenyl radical	1H-indol-5-yl	369
12	Phenyl radical	1H-indazol-6-yl	369
				13	Pyridin-4-yl	1H-indazol-6-yl	370
14	Thiazol-2-yl	1H-indazol-6-yl	376
				15	Phenyl radical	1H-benzotriazol-5-yl	370
16	Pyridin-2-yl	1-methyl-1H-indazol-5-yl	384
				17	2-chlorophenyl group	1H-indazol-6-yl	403
18	4, 5-dimethyl-thiazol-2-yl	1H-indazol-6-yl	404
				19	2, 4-dichlorophenyl	1H-indazol-6-yl	437
20	Benzothiazol-2-yl	1H-indazol-6-yl	426
				21	4-Phenylthiazol-2-yl	1H-indazol-6-yl	452
22	2-fluorophenyl group	1H-indazol-6-yl	387
				23	2-ethylphenyl radical	1H-indazol-6-yl	397
24	2, 4-dichlorophenyl	1H-benzotriazol-5-yl	439

Example 25

Synthesis of 2- (1-isopropyl-1H-imidazol-2-ylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

Coupling of 4-amino-3-nitrobenzoic acid (27mmol) with 6-aminoindazole (30mmol) using HBTU (30mmol) using general procedure D with anhydrous DMF as solvent gave 4-amino-N- (1H-indazol-6-yl) -3-nitrobenzamide which was used for further transformation without further purification.

The nitroaniline described above was reduced to 3, 4-diamino-N- (1H-indazol-6-yl) -benzamide under a hydrogen atmosphere as described in general procedure F.

2-bromopropane (7mmol) and K₂CO₃(13mmol) was added to a solution of 2-nitroimidazole (4mmol) in DMF (10 mL). The mixture was stirred at 60 ℃ for 4 h. The contents were cooled to room temperature, water (20mL) was added, and the mixture was extracted with EtOAc (3X 10 mL). Combining the extracts over MgSO₄Drying, filtering and removing the solvent in vacuo to obtain 1-isopropyl-2-nitro-1H-imidazole. The product was used for further transformation without further purification.

The nitroimidazole described above was reduced to 1-isopropyl-2-amino-1H-imidazole under a hydrogen atmosphere as described in general procedure F. Following general procedure A, aminoimidazole (2mmol) was converted to 1-isopropyl-2-isothiocyanato-1H-imidazole.

Reaction of the isothiocyanate (1mmol) with 3, 4-diamino-N- (1H-indazol-6-yl) -benzamide (1mmol) followed by cyclization with EDC gave 2- (1-isopropyl-1H-imidazol-2-ylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide as described in general procedure B. MS: m/z 401(M + H)⁺。

The compound listed in table 3 was synthesized according to the procedure for example 25 using 3, 4-diamino-N- (1H-indazol-6-yl) -benzamide.

TABLE 3

Examples	Ar	MS(m/z)
			26	2, 4-dimethylphenyl	397
27	2-isopropylphenyl	411
			28	4-chlorophenyl group	403
29	Naphthalen-1-yl	419

Examples	Ar	MS(m/z)
			30	2-tert-butylphenyl	425
31	Biphenyl-2-yl	445
			32	2-propylphenyl	411
33	2, 5-dichlorophenyl	438
			34	2-methoxyphenyl radical	399
35	2-trifluoromethylphenyl group	437
			36	3-methylpyridin-2-yl	384
37	2-trifluoromethoxyphenyl group	453
			38	3-fluorophenyl group	387
39	4-fluorophenyl group	387
			40	3, 5-difluorophenyl	405
41	2-butylphenyl group	425
			42	3-Ethyl-6-methylpyridin-2-yl	412
43	5-chloro-2-methylphenyl radical	417
			44	3-fluoro-2-methylphenyl	411
45	5-fluoro-2-methylphenyl	411
			46	3-chloro-2-methylphenyl	417
47	1-cyclopentyl-1H-imidazol-2-yl	427

Example 48

Synthesis of 2- (2-isopropylphenylamino) -6- (4-methylpiperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

Solid 2-chloro-4-fluorobenzoic acid (10mmol) was added portionwise to a flask containing concentrated sulfuric acid (5mL) with rapid stirring. The reaction mixture was then cooled to 0 ℃ and 70% nitric acid (12mmol) was added dropwise. After the addition was complete, the reaction mixture was allowed to warm to room temperature and stirred for 1-2 h. The reaction mixture was poured into 50g of ice, and the solid was collected by filtration, washed with water, and dried. The product, 2-chloro-4-fluoro-5-nitrobenzoic acid, was used for further conversion without further purification.

Following general procedure E, 2-chloro-4-fluoro-5-nitrobenzoic acid (5mmol) obtained above was converted to the corresponding acid chloride and reacted with 6-aminoindazole (5 mmol). The resulting product, 2-chloro-4-fluoro-N- (1H-indazol-6-yl) -5-nitrobenzamide, was a light orange solid, which was used for further conversion without further purification. MS: m/z 335(M + H)⁺。

2-chloro-4-fluoro-N- (1H-indazol-6-yl) -5-nitrobenzamide (4mmol) obtained above was treated with ammonium hydroxide (4mL) as described in general procedure G to give 4-amino-2-chloro-N- (1H-indazol-6-yl) -5-nitrobenzamide as a yellow solid. MS: m/z 332(M + H)⁺。

Following general procedure H, neat 4-amino-2-chloro-N- (1H-indazol-6-yl) -5-nitrobenzamide (3mmol) obtained above was heated with N-methylpiperazine (5mL) to give 4-amino-N- (1H-indazol-6-yl) -2- (4-methylpiperazin-1-yl) -5-nitrobenzamide. This product was reduced to 4, 5-diamino-N- (1H-indazol-6-yl) -2- (4-methylpiperazin-1-yl) benzamide under hydrogenation conditions as described in general procedure F.

As described in general operation B, withReaction of the diamine (1mmol) obtained above with 1-isopropyl-2-isothiocyanatobenzene (1mmol) followed by cyclization using EDC gave 2- (2-isopropylphenylamino) -6- (4-methylpiperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) amide. MS: m/z 509(M + H)⁺。

The compound listed in table 4 was synthesized according to the procedure for example 48 using 4-amino-2-chloro-N- (1H-indazol-6-yl) -5-nitrobenzamide.

TABLE 4

Examples	Ar	X	MS(m/z)
				49	2-isopropylphenyl	Morpholin-4-yl	496
50	2-trifluoromethylphenyl group	4-methylpiperazin-1-yl	535
				51	3, 5-difluorophenyl	4-methylpiperazin-1-yl	503
52	2, 4-dichlorophenyl	4-methylpiperazin-1-yl	536
				53	Thiazol-2-yl	4-methylpiperazin-1-yl	474
54	2-trifluoromethylphenyl group	Morpholin-4-yl	522
				55	3, 5-difluorophenyl	Morpholin-4-yl	490
56	2, 4-dichlorophenyl	Morpholin-4-yl	522
				57	2-trifluoromethylphenyl group	Piperidin-1-yl radical	520
58	3-methylpyridin-2-yl	4-methylpiperazin-1-yl	482
				59	Thiazol-2-yl	Morpholin-4-yl	461
60	3-methylpyridin-2-yl	Morpholin-4-yl	469
				61	1-isopropyl-1H-imidazol-2-yl	4-methylpiperazin-1-yl	499
62	1-cyclopentyl-1H-imidazol-2-yl	4-methylpiperazin-1-yl	525
				63	1-isopropyl-1H-imidazol-2-yl	Morpholin-4-yl	486
64	1-cyclopentyl-1H-imidazol-2-yl	Morpholin-4-yl	512
				65	2-ethyl-2H-pyrazol-3-yl	Morpholin-4-yl	472

Example 66

Synthesis of 2- (2-isopropylphenylamino) -3H-benzimidazole-5-carboxylic acid [3- (2-morpholin-4-yl-ethylamino) -1H-indazol-6-yl ] -amide

To a solution of 2, 6-dinitro-2H-indazole (1mmol) (prepared by nitration of 6-nitroindazole; Wrzeciono, et al, E.Pharmazie, 1980, 35, 593-propan 596) in anhydrous THF (4mL) at 0 deg.C was added dropwise 2-morpholin-4-yl-ethylamine (2 mmol). The reaction mixture was allowed to warm to room temperature and stirred for 12 h. The contents were diluted with ethyl acetate (20mL), washed with water (2X 10mL) and brine (10mL), and dried over anhydrous sodium sulfate. The solvent was removed in vacuo to give 3- (2-morpholin-4-yl-ethylamino) -6-nitro-1H-indazole as a brown solid which was reduced to 3- (2-morpholin-4-yl-ethylamino) -1H-indazol-6-ylamine by hydrogenation according to general procedure F.

Following general procedure B, 1-isopropyl-2-isothiocyanatobenzene (5mmol) was reacted with methyl 3, 4-diaminobenzoate (5mmol) to give methyl 2- (2-isopropylphenylamino) -1H-benzimidazole-5-carboxylate, which was purified by chromatography on silica gel using DCM/ethyl acetate as eluent.

The ester obtained above is hydrolyzed using general procedure C to give 2- (2-isopropylphenylamino) -1H-benzimidazole-5-carboxylic acid. The carboxylic acid was reacted using HBTU using general procedure D(0.25mmol) was coupled with 3- (2-morpholin-4-yl-ethylamino) -1H-indazol-6-ylamine (0.25 mmol). After purification by silica gel chromatography using DCM/methanol as eluent, the product 2- (2-isopropylphenylamino) -3H-benzimidazole-5-carboxylic acid [3- (2-morpholin-4-yl-ethylamino) -1H-indazol-6-yl]-an amide. MS: m/z 539(M + H)⁺。

The following compounds, as shown in table 5, were synthesized using the procedure described in example 66.

TABLE 5

Examples	R	MS(m/z)
			67	3-morpholin-4-yl-propylamino	553
68	Methylamino radical	440

Example 69

Synthesis of 2- (pyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (3-amino-1H-indazol-6-yl) -amide

To a solution of 2-fluoro-4-nitrobenzonitrile (10mmol) in isopropanol (30mL) was added aqueous hydrazine (4 mL). The resulting solution was heated at 80 ℃ for 12 h. The reaction mixture was then concentrated, water (30mL) was added, and the solution was extracted with ethyl acetate (2X 25 mL). The organic layers were combined, washed with water (30mL) and brine (30mL), and dried over anhydrous sodium sulfate. The volatiles were removed in vacuo to give 3-amino-6-nitroindazole as an orange solid. Used for further transformation without further purification.

Hydrogenation of the nitro compound described above according to general procedure F gave 3, 6-diaminoindazole.

2-Isothiocyanatopyridine (4mmol), prepared from 2-aminopyridine using general procedure A, was reacted with methyl 3, 4-diaminobenzoate as described in general procedure B to give methyl 2- (pyridin-2-ylamino) -1H-benzimidazole-5-carboxylate. This ester was hydrolyzed according to general procedure C to give 2- (pyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid.

The carboxylic acid (0.5mmol) and 3, 6-diaminoindazole (0.5mmol) were coupled using HBTU as described in general procedure D to give 2- (pyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (3-amino-1H-indazol-6-yl) -amide. MS: m/z 385(M + H)⁺。

Example 70

Synthesis of 2- (2-isopropylphenylamino) -3H-benzimidazole-5-carboxylic acid {3- [ (1-methylpiperidine-4-carbonyl) -amino ] -1H-indazol-6-yl } -amide

Using general procedure E, 1-methylpiperidine-4-carbonyl chloride (1mmol), prepared from the corresponding carboxylic acid using general procedure E, was reacted with 3, 6-diaminoindazole (1mmol) (see example 69) to give 1-methylpiperidine-4-carboxylic acid (6-amino-1H-indazol-3-yl) -amide.

Coupling of 2- (2-isopropylphenylamino) -3H-benzimidazole-5-carboxylic acid (0.3 mmol; see example 66) with the above-described 1-methylpiperidine-4-carboxylic acid (6-amino-1H-indazol-3-yl) -amide (0.3mmol) using HBTU affords the desired product 2- (2-isopropylphenylamino) -3H-benzimidazole-5-carboxylic acid {3- [ (1-methylpiperidine-4-carbonyl) -amino ] -as described in general procedure D]-1H-indazol-6-yl } -amide. MS: m/z 551(M + H)⁺。

The following compounds, as shown in table 6, were synthesized using the procedure described in example 70.

TABLE 6

Examples	Ar	R	MS(m/z)
				71	Pyridin-2-yl	Methyl radical	427
72	2, 4-dichlorophenyl	Methyl radical	494
				73	2, 4-dichlorophenyl	Phenyl radical	556

Example 74

Synthesis of 2- (2-isopropylphenylamino) -3H-benzimidazole-5-carboxylic acid (3-methoxy-1H-indazol-6-yl) -amide

To a solution of 2, 6-dinitro-2H-indazole (1mmol) (prepared by nitration of 6-nitroindazole; Wrzeciono, et al, E.Pharmazie, 1980, 35, 593-. The reaction mixture was allowed to warm to room temperature and stirred for 12 h. The contents were diluted with ethyl acetate (20mL), washed with water (2X 10mL) and brine (10mL), and dried over anhydrous sodium sulfate. The solvent was removed in vacuo to give 3-methoxy-6-nitro-1H-indazole as a brown solid, which was reduced by hydrogenation to 3-methoxy-1H-indazol-6-ylamine according to general procedure F.

Coupling of 2- (2-isopropylphenylamino) -3H-benzimidazole-5-carboxylic acid (0.3 mmol; see example 66) with the above 3-methoxy-1H-indazol-6-ylamine (0.3mmol) using HBTU as described in general procedure D gave the desired product 2- (2-isopropylphenylamino) -3H-benzimidazole-5-carboxylic acid (3-methoxy-1H-indazol-6-yl) -amide. MS: m/z 441(M + H)⁺。

Example 75

Synthesis of 2- (2-isopropylphenylamino) -3H-benzimidazole-5-carboxylic acid [3- (2-morpholin-4-ylethoxy) -1H-indazol-6-yl ] -amide

To a solution of 2-morpholin-4-yl-ethanol (3mmol) in anhydrous THF (6mL) at 0 deg.C was added sodium hydride (4 mmol; 60% oil dispersion) in portions. The alcoholate thus formed was reacted with 2, 6-dinitro-2H-indazole (1mmol) as described in example 74 to give 3- (2-morpholin-4-ylethoxy) -6-nitro-1H-indazole as a brown solid which was reduced by hydrogenation to 3- (2-morpholin-4-ylethoxy) -1H-indazol-6-ylamine according to general procedure F.

Coupling of 2- (2-isopropylphenylamino) -3H-benzimidazole-5-carboxylic acid (0.3 mmol; see example 66) with 3- (2-morpholin-4-ylethoxy) -1H-indazol-6-ylamine (0.3mmol) as described in general procedure D using HBTU gave the desired product 2- (2-isopropylphenylamino) -3H-benzimidazole-5-carboxylic acid [3- (2-morpholin-4-ylethoxy) -1H-indazol-6-yl ] -amine]-an amide. MS: m/z 540(M + H)⁺。

Example 76

Synthesis of 2- (2, 4-dichlorophenylamino) -3H-benzimidazole-5-carboxylic acid (3-morpholin-4-ylmethyl-1H-indazol-6-yl) -amide

To a solution of 6-nitro-1H-indazole-3-carbaldehyde (0.5 mmol; prepared from 6-nitroindole; Zhang et al, J.Med.chem.2001, 44, 1021-1024) in anhydrous THF (1mL) at room temperature was added morpholine (1mmol) and acetic acid (2 drops) and the mixture was stirred for 1H. The reaction mixture was washed with solid NaCNBH₃(2mmol) and stirringStirring for another 4 h. The contents were poured into water and extracted with ethyl acetate (2X 10 mL). The organic layers were combined and washed with saturated NaHCO₃The aqueous solution (10mL) and brine (10mL) were washed and dried over anhydrous sodium sulfate. The solvent was removed in vacuo to give the desired product 3- (morpholin-4-yl) methyl-6-nitro-1H-indazole.

Hydrogenation of the nitro compound followed general procedure F gave 3- (morpholin-4-yl) methyl-1H-indazol-6-ylamine.

Following general procedure B, 2, 4-dichloro-1-isothiocyanatobenzene (5mmol) was reacted with methyl 3, 4-diaminobenzoate (5mmol) to give methyl 2- (2, 4-dichlorophenylamino) -3H-benzimidazole-5-carboxylate, which was purified by chromatography on silica gel using DCM/ethyl acetate as eluent.

The ester obtained above was hydrolyzed using general procedure C to give 2- (2, 4-dichlorophenylamino) -3H-benzimidazole-5-carboxylic acid. Using general procedure D, the carboxylic acid (0.25mmol) was coupled with 3- (morpholin-4-yl) methyl-1H-indazol-6-ylamine (0.25mmol) using HBTU. After purification by silica gel chromatography using DCM/methanol as eluent, the product 2- (2, 4-dichlorophenylamino) -3H-benzimidazole-5-carboxylic acid (3-morpholin-4-ylmethyl-1H-indazol-6-yl) -amide is obtained as a light brown solid. MS: m/z 536(M + H)⁺。

Example 77

Synthesis of 2- (2, 4-dichlorophenylamino) -3H-benzimidazole-5-carboxylic acid (3-methyl-1H-indazol-6-yl) -amide

To a solution of 6-nitro-1H-indazole-3-carbaldehyde (0.5 mmol; prepared from 6-nitroindole; Zhanget al, J.Med.chem.2001, 44, 1021-. The reaction mixture was neutralized to pH-7 with acetic acid, concentrated in vacuo, and concentrated withDiluted with water and extracted with ethyl acetate (3X 8 mL). The organic layers were combined and washed with saturated NaHCO₃The aqueous solution (10mL) and brine (10mL) were washed and dried over anhydrous sodium sulfate. The solvent was removed in vacuo to afford the desired product, 3-methyl-1H-indazol-6-ylamine.

Using general procedure D, the amine obtained above (0.25mmol) was coupled with 2- (2, 4-dichloro-phenylamino) -3H-benzimidazole-5-carboxylic acid (0.25 mmol; cf. example 76) using HBTU. After purification by silica gel chromatography using DCM/methanol as eluent, the product 2- (2, 4-dichlorophenylamino) -3H-benzimidazole-5-carboxylic acid (3-methyl-1H-indazol-6-yl) -amide was obtained as a light brown solid. MS: m/z 452(M + H)⁺。

Example 78

Synthesis of 2- (2-ethylphenylamino) -3H-benzimidazole-5-carboxylic acid (3-chloro-1H-indazol-6-yl) -amide

To a solution of 6-nitroindazole (2mmol) in DCE (5mL) was added sulfonyl chloride (10mmol) and the resulting mixture was heated at 80 ℃ for 3-5 h. The reaction mixture was concentrated and 5% Na was added₂CO₃Aqueous (20mL) and extracted with EtOAc (2X 15 mL). The organic layers were combined and washed with water (20mL) and brine (20mL) over anhydrous Na₂SO₄And (5) drying. The volatiles were removed to give 3-chloro-6-nitro-1H-indazole as a yellow solid.

To a solution of the above nitro compound (0.5mmol) in methanol (2mL) was added solid sodium bisulfite (3mmol) and concentrated ammonium hydroxide (0.25 mL). The resulting mixture was stirred at room temperature for 12 h. The contents were filtered through celite and the solvent was removed in vacuo. The resulting residue was purified by chromatography on silica gel using ethyl acetate/hexane as eluent to give 3-chloro-1H-indazol-6-ylamine as a light brown solid.

Following general procedure B, 2-ethyl-1-isothiocyanatobenzene (3mmol) was reacted with methyl 3, 4-diaminobenzoate (3mmol) to give methyl 2- (2-ethylphenylamino) -3H-benzimidazole-5-carboxylate, which was purified by silica gel chromatography using DCM/ethyl acetate as eluent.

The ester obtained above was hydrolyzed using general procedure C to give 2- (2-ethylphenylamino) -3H-benzimidazole-5-carboxylic acid. Using general procedure D, the carboxylic acid (0.25mmol) was coupled with 3- (morpholin-4-yl) methyl-1H-indazol-6-ylamine (0.25mmol) using HBTU. After purification by silica gel chromatography using DCM/methanol as eluent, the product 2- (2-ethylphenylamino) -3H-benzimidazole-5-carboxylic acid (3-chloro-1H-indazol-6-yl) -amide was obtained as a light brown solid. MS: m/z 431(M + H)⁺。

Example 79

Synthesis of tert-butyl 2- [6- (1H-indazol-6-ylcarbamoyl) -1H-benzimidazol-2-ylamino ] -6, 7-dihydro-4H-thiazolo [5, 4-c ] pyridine-5-carboxylate

To a solution of 1-Boc-4-piperidone (5mmol) in anhydrous THF (20mL) was added solid Ba₂CO₃(10 mmol). The resulting mixture was stirred vigorously. The reaction mixture was treated with pyrrolidone trihydrobromide (5.5mmol) in portions at room temperature. After 3h, the contents were filtered and the solvent was removed. The crude reaction mixture containing the product, 3-bromo-4-oxo-piperidine-1-carboxylic acid tert-butyl ester, was used for further transformations without further purification.

To a solution of the bromo compound (5mmol) obtained above in acetone (20mL) was added solid thiourea (6mmol) and solid K₂CO₃(10mmol) the reaction mixture was stirred at room temperature for 12 h. BOC anhydride (5mmol) was added to the reaction mixture and the reaction stirred for 4 h. The contents were then filtered to remove the solvent. The resulting residue was purified by chromatography on silica gel using DCM/methanol as eluent. The product 2-amino-6 is obtained,7-dihydro-4H-thiazolo [5, 4-c]Pyridine-5-carboxylic acid tert-butyl ester as a pale yellow solid.

Using general procedure A, the above amine (0.5mmol) was converted to the corresponding isothiocyanate and reacted with 3, 4-diamino-N- (1H-indazol-6-yl) -benzamide (0.5 mmol; see example 25) according to general procedure B to give 2- [6- (1H-indazol-6-ylcarbamoyl) -1H-benzimidazol-2-ylamino-)]-6, 7-dihydro-4H-thiazolo [5, 4-c]Pyridine-5-carboxylic acid tert-butyl ester. MS: m/z 531(M + H)⁺。

Example 80

Synthesis of 2- (4, 5, 6, 7-tetrahydrothiazolo [5, 4-c ] pyridin-2-ylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

To 2- [6- (1H-indazol-6-ylcarbamoyl) -1H-benzimidazol-2-ylamino]-6, 7-dihydro-4H-thiazolo [5, 4-c]A solution of pyridine-5-carboxylic acid tert-butyl ester (0.25 mmol; see example 79) in methanol (1mL) was added 4M HCl in dioxane (0.5 mL). The resulting mixture was stirred at room temperature for 5-6 h. The volatiles were removed in vacuo and the resulting residue was suspended in diethyl ether. The resulting solid was collected by filtration, washed with diethyl ether and dried in vacuo to give 2- (4, 5, 6, 7-tetrahydro-thiazolo [5, 4-c ]]Pyridin-2-ylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide as the hydrochloride salt. MS: m/z 431(M + H)⁺。

Example 81

Synthesis of 2- (2-isopropylphenylamino) -3H-benzimidazole-5-carboxylic acid [1- (2-hydroxy-ethyl) -1H-indazol-5-yl ] -amide

To a solution of 2-chloro-5-nitrobenzaldehyde (4mmol) in ethanol (10mL) was added aqueous hydrazine (5mmol) and the resulting solution was heated at reflux for 2h to complete the formation of the hydrazone. DIEA (10mmol) was added to the reaction mixture and the reaction was subjected to microwave irradiation at 150 ℃ for 8-10 h. After removal of volatiles in vacuo, the resulting residue was dissolved in EtOAc (30mL), washed with water (20mL) and brine (20mL), and dried over anhydrous sodium sulfate. The solvent was removed in vacuo to give the product 2- (5-nitroindazol-1-yl) -ethanol.

Reduction of the nitro compound under hydrogenation conditions as described in general procedure F gave 2- (5-aminoindazol-1-yl) -ethanol. Aminoindazole (0.3mmol) and 2- (2-isopropylphenylamino) -1H-benzimidazole-5-carboxylic acid (0.3 mmol; see example 66) were coupled using HBTU to give 2- (2-isopropylphenylamino) -3H-benzimidazole-5-carboxylic acid [1- (2-hydroxy-ethyl) -1H-indazol-5-yl ] -2- (2-isopropylphenylamino) -3H-benzimidazole-5-carboxylate as described in general procedure D]-an amide. MS: m/z 455(M + H)⁺。

Example 82

Synthesis of 2- (2-cyclohexylphenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

To a solution of 1-bromo-2-cyclohexyl-benzene (5mmol) in dioxane (20mL) was added solid Pd (OAc)₂(0.1g) and solid CsCO₃(10 mmol). Tert-butyl carbamate (7mmol) was added to the reaction mixture and the contents were heated at 80 ℃ for 2 h. The reaction mixture was cooled to room temperature and filtered through celite. The solvent was removed in vacuo and the resulting residue was purified by flash column chromatography using DCM as eluent to give (2-cyclohexylphenyl) -carbamic acid tert-butyl ester.

Following the procedure described for example 80, the carbamate obtained above was treated with 4M HCl in dioxane to give 2-cyclohexylphenylamine as the hydrochloride salt.

To a solution of the above amine hydrochloride (1mmol) in anhydrous DMF (2mL) was added DIEA (1.5mmol) and 1, 1' -thiocarbonylimidazole (1 mmol). The reaction mixture was heated at 70 ℃ for 1h to give 1-cyclohexyl-2-isothiocyanatobenzene as described in general procedure A.

Following general procedure B, isothiocyanate (0.5mmol) was reacted with 3, 4-diamino-N- (1H-indazol-6-yl) -benzamide (0.5 mmol; see example 25) to give 2- (2-cyclohexylphenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide. MS: m/z 451(M + H)⁺。

Example 83

Synthesis of 2- (3-methylthiophen-2-ylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

To a solution of 3-methylthiophene-2-carboxylic acid (7mmol) in anhydrous dioxane (20mL) was added diphenylphosphoryl azide (7mmol), tert-butanol (6mL) and TEA (1 mL). The resulting mixture was stirred at reflux for 16 h. The reaction mixture was cooled to room temperature and washed with H₂Diluted O (40mL) and extracted with EtOAc (3X 20 mL). The combined extracts were dried (MgSO)₄) The solvent was removed in vacuo. The resulting residue was purified by flash column chromatography using hexane/EtOAc (7: 3) as eluent to give (3-methylthiophen-2-yl) -carbamic acid tert-butyl ester.

To a solution of the carbamate (3mmol) obtained above in dry DCM (10mL) was added a 4MHCl solution in dioxane (8 mL). The mixture was stirred at room temperature for 2 h. The solvent was removed in vacuo. The resulting solid was taken up in anhydrous Et₂O wash (3X 10mL) and dry under reduced pressure to give 3-methylthiophen-2-ylamine as the hydrochloride salt.

To a solution of the above amine hydrochloride (1mmol) in anhydrous DMF (2mL) was added DIEA (1.5mmol) and 1, 1' -thiocarbonylimidazole (1 mmol). The reaction mixture was heated at 70 ℃ for 1h to give 1-cyclohexyl-2-isothiocyanatobenzene as described in general procedure A.

Following general procedure B, isothiocyanate (0.5mmol) was reacted with 3, 4-diamino-N- (1H-indazol-6-yl) -benzamide (0.5 mmol; see example 25) to give 2- (2-cyclohexylphenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide. MS: m/z 389(M + H)⁺。

Example 84

Synthesis of 1H-indazole-6-carboxylic acid [2- (2-isopropylphenylamino) -3H-benzimidazol-5-yl ] -amide

To a solution of 2-chloro-5-nitro-1H-benzimidazole (1.5 mmol; prepared from nitration of 2-chloro-1H-benzimidazole; Galy et al, J.heterocyclic. chem.1997, 34, 6, 1781-1788) in anhydrous NMP (3mL) was added 2-isopropylaniline (4 mmol). The resulting solution was subjected to microwave irradiation at 150 ℃ for 1 h. The contents were cooled to room temperature, diluted with water (20mL), and extracted with EtOAc (2X 15 mL). The extracts were combined, then washed with water (20mL) and brine (20mL), and dried over anhydrous sodium sulfate. The solvent was removed in vacuo and the resulting residue was purified by silica gel chromatography using EtOAc/hexanes as the eluent to give (2-isopropylphenyl) - (5-nitro-1H-benzimidazol-2-yl) -amine as a light yellow solid.

Reduction of the nitro compound (1mmol) under hydrogenation conditions as described in general procedure F gave N²- (2-isopropylphenyl) -1H-benzimidazole-2, 5-diamine.

Methylindazole-6-carboxylic acid ester (4 mmol; Batt et al, J.Med.Chem.2000, 43, 41-58) was hydrolyzed as in general procedure C to give 1H-indazole-6-carboxylic acid. The carboxylic acid (0.5mmol) was reacted with HBTU as described in general procedure DN²Coupling of- (2-isopropylphenyl) -1H-benzimidazole-2, 5-diamine (0.5mmol) gave 1H-indazole-6-carboxylic acid [2- (2-isopropylphenylamino) -3H-benzimidazol-5-yl]Amide as off-white solid. MS: m/z 411(M + H)⁺。

Example 85

Synthesis of 6- (4-methylpiperazin-1-yl) -2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-5-yl) -amide

Following the procedure described for example 48, 2-chloro-4-fluoro-N- (1H-indazol-5-yl) -5-nitrobenzamide was obtained from 2-chloro-4-fluoro-5-nitrobenzoic acid (5mmol) and 5-aminoindazole (5 mmol). The product was obtained as a yellow solid and used for further conversion without further purification. MS: m/z 335(M + H)⁺。

2-chloro-4-fluoro-N- (1H-indazol-5-yl) -5-nitrobenzamide (4mmol) obtained above was treated with ammonium hydroxide (4mL) as described in general procedure G to give 4-amino-2-chloro-N- (1H-indazol-5-yl) -5-nitrobenzamide as a yellow solid. MS: m/z 332(M + H)⁺。

Following the procedure of example 48, the above 4-amino-2-chloro-N- (1H-indazol-5-yl) -5-nitrobenzamide (3mmol) was reacted with N-methylpiperazine (5 mL). The product formed by reduction under hydrogenation conditions was 4, 5-diamino-N- (1H-indazol-5-yl) -2- (4-methylpiperazin-1-yl) benzamide as described in general procedure F.

Reaction of diamine (1mmol) obtained above with 1-trifluoromethyl-2-isothiocyanatobenzene (1mmol) followed by cyclization with EDC gave 2- (2-trifluoromethylphenylamino) -6- (4-methylpiperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-5-yl) amide as described in general procedure B. MS: m/z 535(M + H)⁺。

Example 86

Synthesis of 6-morpholin-4-yl-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-5-yl) -amide

Following procedure G, 4-amino-2-chloro-N- (1H-indazol-5-yl) -5-nitrobenzamide (3 mmol; see example 85) was reacted with morpholine (5 mL). The product formed by reduction under hydrogenation conditions was 4, 5-diamino-N- (1H-indazol-5-yl) -2-morpholin-4-yl-benzamide as described in general procedure F.

Reaction of diamine (1mmol) obtained above with 1-trifluoromethyl-2-isothiocyanatobenzene (1mmol) followed by cyclization with EDC gave 6-morpholin-4-yl-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-5-yl) -amide as described in general procedure B. MS: m/z 522(M + H)⁺。

Example 87

Synthesis of 4- [6- (1H-indazol-6-ylcarbamoyl) -2- (2-trifluoromethylphenylamino) -3H-benzimidazol-5-yl ] -piperazine-1-carboxylic acid tert-butyl ester

Following general procedure H, a solution of 4-amino-2-chloro-N- (1H-indazol-6-yl) -5-nitrobenzamide (3 mmol; see example 48) in dioxane (5mL) was reacted with piperazine (9mmol) to give 4-amino-N- (1H-indazol-6-yl) -5-nitro-2-piperazin-1-yl-benzamide. The product was dissolved in anhydrous THF (6mL), treated with BOC anhydride (3.6mmol) and stirred for 4-6 h. The solvent was removed to dryness and the resulting residue was suspended in diethyl ether (50mL) with stirring. The resulting solid was collected by filtration, washed with diethyl ether and dried in vacuo to give 4- [ 5-amino-2- (1H-indazol-6-ylcarbamoyl) -4-nitro-phenyl ] -piperazine-1-carboxylic acid tert-butyl ester.

To a solution of the above nitro compound (1mmol) in methanol (4mL) was added solid sodium bisulfite (4mmol) and concentrated ammonium hydroxide (0.5 mL). The resulting mixture was heated at reflux for 5-8 h. The reaction was concentrated and the residue dissolved in THF (20mL) with vigorous stirring. The contents were then filtered through celite and the solvent removed in vacuo to give tert-butyl 4- [4, 5-diamino-2- (1H-indazol-6-ylcarbamoyl) -phenyl ] piperazine-1-carboxylate, which was used for further conversion without further purification.

Reaction of the above diamine (0.3mmol) with 1-trifluoromethyl-2-isothiocyanatobenzene (0.3mmol) followed by cyclization with EDC gave 4- [6- (1H-indazol-6-ylcarbamoyl) -2- (2-trifluoromethylphenylamino) -3H-benzimidazol-5-yl]-piperazine-1-carboxylic acid tert-butyl ester. MS: m/z 621(M + H)⁺。

Example 88

Synthesis of 6-piperazin-1-yl-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

The product of example 87 was treated with 4M HCl in dioxane using the procedure described for example 80 to give 6-piperazin-1-yl-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide as the hydrochloride salt. MS: m/z 521(M + H)⁺。

Example 89

Synthesis of tert-butyl 4- [6- (1H-indazol-6-ylcarbamoyl) -2- (3-methylpyridin-2-ylamino) -3H-benzimidazol-5-yl ] -piperazine-1-carboxylate

4- [4, 5-diamino-2- (1H-indazol-6-ylcarbamoyl) -phenyl ] as described in general procedure B]Reaction of piperazine-1-carboxylic acid tert-butyl ester (see example 87) with 2-isothiocyanato-3-methylpyridine (0.3 mmol; prepared from 3-methylpyridin-2-ylamine according to general procedure A), followed by cyclization with EDC gave 4- [6- (1H-indazol-6-ylcarbamoyl) -2- (3-methylpyridin-2-ylamino) -3H-benzimidazol-5-yl]-piperazine-1-carboxylic acid tert-butyl ester. MS: m/z568(M + H)⁺。

Example 90

Synthesis of 2- (3-methylpyridin-2-ylamino) -6-piperazin-1-yl-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

The product of example 89 was treated with 4M HCl in dioxane using the procedure described for example 80 to give 2- (3-methylpyridin-2-ylamino) -6-piperazin-1-yl-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide as the hydrochloride salt. MS: m/z 468(M + H)⁺。

Example 91

Synthesis of 2- (2, 6-diethylphenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

Following general procedure B, a 1: 1DMF/THF (2mL) solution of 1, 3-diethyl-2-isothiocyanatobenzene (0.5mmol) was reacted with 3, 4-diamino-N- (1H-indazol-6-yl) -benzamide (0.5 mmol; see example 25) to give 2- (2, 6-diethylphenylamino)-3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide. MS: m/z 425(M + H)⁺。

Example 92

Synthesis of 6-diisobutylamino-2- (2-trifluoromethylphenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

To a solution of 4-amino-2-chloro-N- (1H-indazol-6-yl) -5-nitrobenzamide (1 mmol; see example 48) in NMP (2mL) was added diisobutylamine (0.5mL) and the resulting mixture was subjected to microwave irradiation at 140 ℃ for 1H. The reaction mixture was cooled to room temperature and diluted with water (20 mL). The resulting solid was collected by filtration, washed with water, and dried in vacuo to give 4-amino-2-diisobutylamino-N- (1H-indazol-6-yl) -5-nitrobenzamide.

The nitro compound (0.5mmol) was reduced under hydrogenation conditions as described in general procedure F to give 4, 5-diamino-2-diisobutylamino-N- (1H-indazol-6-yl) benzamide.

Reaction of the above diamine (0.3mmol) with 1-trifluoromethyl-2-isothiocyanatobenzene (0.3mmol) followed by cyclization with EDC gave 6-diisobutylamino-2- (2-trifluoromethylphenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide as described in general procedure B. MS: m/z 564(M + H)⁺。

Example 93

Synthesis of 6-diethylamino-2- (3-methylpyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

To a solution of 4-amino-2-chloro-N- (1H-indazol-6-yl) -5-nitrobenzamide (1 mmol; see example 48) in NMP (2mL) was added diethylamine (1.0mL) and the resulting mixture was subjected to microwave irradiation at 70 ℃ for 1H. The reaction mixture was cooled to room temperature and diluted with water (20 mL). The resulting solid was collected by filtration, washed with water, and dried in vacuo to give 4-amino-2-diethylamino-N- (1H-indazol-6-yl) -5-nitrobenzamide.

Reduction of the nitro compound (0.5mmol) under hydrogenation conditions as described in general procedure F gave 4, 5-diamino-2-diethylamino-N- (1H-indazol-6-yl) benzamide.

Reaction of the above diamine (0.3mmol) with 2-isothiocyanato-3-methylpyridine (0.3 mmol; prepared from 3-methylpyridin-2-ylamine according to general procedure a) followed by cyclization with EDC gave 6-diethylamino-2- (3-methyl-pyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide as described in general procedure B. MS: m/z 455(M + H)⁺。

The compound listed in Table 7 was synthesized according to the procedure for example 93, using 4-amino-2-chloro-N- (1H-indazol-6-yl) -5-nitrobenzamide.

TABLE 7

Examples	Ar	X	MS(m/z)
				94	2-trifluoromethylphenyl group	2, 6-dimethylmorpholin-4-yl	550
95	2-trifluoromethylphenyl group	Diethylamino group	508
				96	2-trifluoromethylphenyl group	(2-dimethylaminoethyl) methylamino	537
97	2-trifluoromethylphenyl group	4-dimethylaminopiperidin-1-yl	563
				98	2-trifluoromethylphenyl group	Dipropylamino group	536
99	3-methylpyridin-2-yl	Dipropylamino group	483
				100	2-trifluoromethylphenyl group	Bis- (2-methoxyethyl) amino	568
101	2-trifluoromethylphenyl group	4-hydroxypiperidin-1-yl	536
				102	2-trifluoromethylphenyl group	Ethyl- (2-methoxyethyl) amino	538
103	3-methylpyridin-2-yl	Bis- (2-methoxyethyl) amino	515
				104	3-methylpyridin-2-yl	Pyrrolidin-1-yl radical	453
105	2-trifluoromethylphenyl group	Pyrrolidin-1-yl radical	506
				106	2-trifluoromethylphenyl group	(2-dimethylaminoethyl) ethylamino	551
107	3-methylpyridin-2-yl	4-hydroxypiperidin-1-yl	483
				108	3-methylpyridin-2-yl	Ethyl- (2-methoxyethyl) amino	485
109	2-trifluoromethylphenyl group	Ethyl propyl amino	522
				110	3-methylpyridin-2-yl	Ethyl propyl amino	469
111	2-trifluoromethylphenyl group	4-isopropylpiperazin-1-yl	563
				112	2-trifluoromethylphenyl group	Ethyl methyl amino group	494
113	3-methylpyridin-2-yl	Ethyl methyl amino group	441
				114	3-methylpyridin-2-yl	4-isopropylpiperazin-1-yl	510

Example 115

Synthesis of 2- (3-chloropyridin-2-ylamino) -6-diethylamino-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

To a suspension of 2-chloro-4-fluoro-5-nitrobenzoic acid (5mmol) in dry DMF (0.2mL) in dry DCM (5mL) was added oxalyl chloride (15mmol) and the mixture was stirred at 50 ℃. After completion of the reaction (-60 min), the solvent was removed in vacuo to afford the acid chloride. Toluene (. about.1 mL) was added to the acid chloride and the solvent was removed to dryness in vacuo to ensure complete removal of residual oxalyl chloride. The product, 4-chloro-2-fluoro-5-nitrobenzoyl chloride, was obtained as a pale yellow solid.

The acid chloride (. about.5 mmol) obtained above was dissolved in EtOAc (5mL) and added dropwise to a suspension of 6-aminoindazole (4.5mmol) in EtOAc (15mL) containing triethylamine (1mL) at 0-5 ℃. The mixture was then allowed to warm to room temperature and stirred for 2-3 h. Most of the solvent was removed in vacuo and hexane was added to the residue. The solid was collected on the filter, washed twice with hexane/EtOAc (5: 1) and three times with water. The residue was dried in vacuo to give the product 2-chloro-4-fluoro-N- (1H-indazol-6-yl) -5-nitrobenzamide as a yellow solid which was used for further conversion without further purification. MS: m/z 335(M + H)⁺。

To a solution of 2-chloro-4-fluoro-N- (1H-indazol-6-yl) -5-nitrobenzamide (3mmol) in dioxane (6mL) was added concentrated NH₄Aqueous OH (3 mL). The resulting mixture was heated at 60 ℃ for 2-3 h. After the reaction is finished, the product 2-amino-4-fluoro-N- (1H-indazol-6-yl) -5-nitrobenzamide is obtained. To the crude reaction mixture was added diethylamine (45 mmol). The mixture was then heated at 60 ℃ for 6 h. After the reaction was complete, the volatiles were removed in vacuo and the residue was suspended in cold water. The solid was collected by filtration, washed with water and dried in vacuo to give 4-amino-2-diethylamino-N- (1H-indazol-6-yl) -5-nitrobenzamide.

The nitro compound (2mmol) obtained above was reduced under hydrogenation conditions as described in general procedure F to give 4, 5-diamino-2-diethylamino-N- (1H-indazol-6-yl) benzamide.

To stirred 2-amino-3-chloropyridine (2mmol) in CHCl at 0 deg.C₃(5mL) solution 0.7M aqueous sodium bicarbonate was added. Thiophosgene (2.2mmol) was added dropwise at 0 ℃ and the contents were gradually warmed to RT over 2 h. The reaction mixture was diluted with DCM (20mL) and the layers were separated. The organic layer was washed with water (2X 10mL) followed by brine (10mL) over anhydrous Na₂SO₄And (5) drying. The volatiles were removed in vacuo and the product, 3-chloro-2-isothiocyanatopyridine, was used without any purification.

Reaction of the above diamine (0.3mmol) with 3-chloro-2-isothiocyanatopyridine (0.3mmol) followed by in situ cyclization using EDC gave 2- (3-chloropyridin-2-ylamino) -6-diethylamino-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide as described in general procedure B. MS: m/z 475(M + H)⁺。

The compounds listed in table 8 were synthesized according to the procedure for example 115 using 4, 5-diamino-2-diethylamino-N- (1H-indazol-6-yl) benzamide.

TABLE 8

Example 133

Synthesis of 6-diethylamino-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide

6-aminobenzothiazole (4.5mmol) was reacted with 4-chloro-2-fluoro-5-nitrobenzoyl chloride (5mmol) using the conditions described for example 115. The reaction mixture was diluted with EtOAc (40mL), washed with water (2X 40mL) and brine (40mL), over anhydrous Na₂SO₄And (5) drying. The organic layer was removed to give the product N-benzothiazol-6-yl-2-chloro-4-fluoro-5-nitrobenzamide as a yellow solid. MS: m/z 352(M + H)⁺。

Using the one-pot procedure described in example 115, a solution of the above amide (3mmol) in dioxane was reacted with NH₄Aqueous OH followed by diethylamine gave 4-amino-N-benzothiazol-6-yl-2-diethylamino-5-nitrobenzamide as a yellow solid.

The nitro compound (2mmol) obtained above was reduced under hydrogenation conditions as described in general procedure F to give 4, 5-diamino-N-benzothiazol-6-yl-2-diethylaminobenzamide.

Reaction of the above diamine (0.3mmol) with 1-isothiocyanic acid as described in general procedure BAcyl-2-trifluoromethylbenzene (0.3mmol) was reacted followed by in situ cyclization using EDC to give 6-diethylamino-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide. MS: m/z 525(M + H)⁺。

The compounds listed in Table 9 were synthesized by the procedure of example 133 using 4, 5-diamino-N-benzothiazol-6-yl-2-diethylaminobenzamide.

TABLE 9

Examples	Ar	MS(m/z)
			134	Bicyclo [2.2.1]Hept-2-yl	475
135	Isopropyl group	423
			136	2, 5-difluorophenyl	493
137	3，5-Difluorophenyl group	493
			138	2, 4-dichlorophenyl	526
139	2-trifluoromethoxyphenyl group	541
			140	2-isopropylphenyl	499

Example 141

Synthesis of 6- (4-methyl-piperazin-1-yl) -2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide

A solution of N-benzothiazol-6-yl-2-chloro-4-fluoro-5-nitrobenzamide (2mmol) in dioxane (4mL) was reacted with NH using the conditions described in example 115₄And (4) reacting an OH aqueous solution. After completion of the formation of 4-amino-N-benzothiazol-6-yl-2-chloro-5-nitrobenzamide, N-methylpiperazine (12mmol) was added to the reaction mixture. The contents were heated at reflux for 10h and the reaction mixture was cooled to RT. The contents were poured onto ice-cold water with vigorous stirring. The resulting solid was collected by filtration, washed with water and dried in vacuo to give the product 4-amino-N-benzothiazol-6-yl-2- (4-methyl-piperazin-1-yl) -5-nitrobenzamide as a yellow solid.

The nitro compound (2mmol) obtained above was reduced under hydrogenation conditions as described in general procedure F to give 4, 5-diamino-N-benzothiazol-6-yl-2- (4-methyl-piperazin-1-yl) -benzamide.

Reaction of the above diamine (0.3mmol) with 1-isothiocyanato-2-trifluoromethylbenzene (0.3mmol) followed by in situ cyclization using EDC gave 6- (4-methyl-piperazin-1-yl) -2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide as described in general procedure B. MS: m/z 552(M + H)⁺。

The compounds listed in Table 10 were synthesized by the procedure of example 141 using 4-amino-N-benzothiazol-6-yl-2-chloro-5-nitrobenzamide.

Watch 10

Examples	Ar	R	MS(m/z)
				142	3-methylpyridin-2-yl	4-methylpiperazin-1-yl	499
143	2-trifluoromethylphenyl group	Morpholino-4-yl	539
				144	3-methylpyridin-2-yl	Morpholino-4-yl	486

Example 145

Synthesis of 6- (3, 5-dimethylpiperazin-1-yl) -2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

A solution of 2-chloro-4-fluoro-N- (1H-indazol-6-yl) -5-nitrobenzamide (1mmol) in dioxane (2mL) was reacted with NH using the conditions described in example 115₄And (4) reacting an OH aqueous solution. After completion of the formation of 2-amino-4-fluoro-N- (1H-indazol-6-yl) -5-nitrobenzamide, 2, 6-dimethylpiperazine (6mmol) was added to the reaction mixture. The contents were heated at reflux for 10h and the reaction mixture was cooled to RT. The contents were poured onto ice-cold water with vigorous stirring. The resulting solid was collected by filtration, washed with water and dried in vacuo to give the product 4-amino-2- (3, 5-dimethyl-piperazin-1-yl) -N- (1H-indazol-6-yl) -5-nitrobenzamide as a yellow solid.

The nitro compound (0.6mmol) obtained above was reduced under hydrogenation conditions as described in general procedure F to give 4, 5-diamino-2- (3, 5-dimethylpiperazin-1-yl) -N- (1H-indazol-6-yl) -benzamide.

Reaction of the above diamine (0.3mmol) with 1-isothiocyanato-2-trifluoromethylbenzene (0.3mmol) followed by in situ cyclization using EDC gave 6- (3, 5-dimethylpiperazin-1-yl) -2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide as described in general procedure B. MS: m/z 549(M + H)⁺。

The compounds listed in Table 11 were synthesized according to the procedure for example 145 using 2-amino-4-fluoro-N- (1H-indazol-6-yl) -5-nitrobenzamide.

TABLE 11

Examples	Ar	R	MS(m/z)
				146	2-trifluoromethylphenyl group	2-methoxyethylamino group	510
147	3-methylpyridin-2-yl	2-methoxyethylamino group	457
				148	2-trifluoromethylbenzyl group	4-methylpiperazin-1-yl	549
149	Benzyl radical	4-methylpiperazin-1-yl	481
				150	Cyclohexyl methyl radical	4-methylpiperazin-1-yl	487
151	Cyclopentyl group	4-methylpiperazin-1-yl	459
				152	(1S, 2S, 4R) -bicyclo [2.2.1]Hept-2-yl	4-methylpiperazin-1-yl	485
153	Adamantan-1-yl radical	4-methylpiperazin-1-yl	525

Example 154

Synthesis of 6-propylamino-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl-amide)

A solution of 2-chloro-4-fluoro-N- (1H-indazol-6-yl) -5-nitrobenzamide (2mmol) in dioxane (4mL) was reacted with NH using the conditions described in example 115₄And (4) reacting an OH aqueous solution. After complete formation of 2-amino-4-fluoro-N- (1H-indazol-6-yl) -5-nitrobenzamide, the volatiles were removed in vacuo. The resulting residue was suspended in cold water while stirring. The resulting solid was collected by filtration, washed with water and dried in vacuo to give the product 4-amino-2-chloro-N- (1H-indazol-6-yl) -5-nitrobenzamide as a yellow solid.

To a solution of the product obtained above (0.5mmol) in NMP (1mL) was added propylamine (0.5 mL). The contents were subjected to microwave irradiation at 80 ℃ for 60 min. The reaction mixture was cooled to room temperature and diluted with water (10 mL). The resulting solid was collected by filtration, washed with water, and dried in vacuo to give 4-amino-N- (1H-indazol-6-yl) -5-nitro-2-propylaminobenzamide.

The nitro compound (0.4mmol) obtained above was reduced under hydrogenation conditions as described in general procedure F to give 4, 5-diamino-N- (1H-indazol-6-yl) -2-propylamino-benzamide.

Reaction of the above diamine (0.3mmol) with 1-isothiocyanato-2-trifluoromethylbenzene (0.3mmol) followed by in situ cyclization using EDC gave 6-propylamino-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) amide as described in general procedure B. MS: m/z 494(M + H)⁺。

Example 155

Synthesis of {1- [6- (1H-indazol-6-ylcarbamoyl) -2- (2-trifluoromethyl-phenylamino) -3H-benzimidazol-5-yl ] -piperidin-4-yl } -carbamic acid tert-butyl ester

To a solution of 4-amino-2-chloro-N- (1H-indazol-6-yl) -5-nitrobenzamide (1 mmol; see example 154) in NMP (2mL) was added piperidin-4-yl-carbamic acid tert-butyl ester (4 mmol). The resulting mixture was heated at 100 ℃ for 10 h. The reaction mixture was cooled to room temperature and diluted with water (20 mL). The resulting solid was collected by filtration, washed with water and dried in vacuo. The crude product was purified by silica gel column chromatography using EtOAc hexanes as eluent to give {1- [ 5-amino-2- (1H-indazol-6-ylcarbamoyl) -4-nitro-phenyl ] -piperidin-4-yl } -carbamic acid tert-butyl ester as a light yellow solid.

The nitro compound (0.5mmol) was reduced under hydrogenation conditions as described in general procedure F to give {1- [4, 5-diamino-2- (1H-indazol-6-ylcarbamoyl) -phenyl ] -piperidin-4-yl } -carbamic acid tert-butyl ester.

Reaction of the above diamine (0.3mmol) with 1-isothiocyanato-2-trifluoromethylbenzene (0.3mmol) followed by in situ cyclization with EDC gave {1- [6- (1H-indazol-6-ylcarbamoyl) -2- (2-trifluoromethyl-phenylamino) -3H-benzimidazol-5-yl]-piperidin-4-yl } -carbamic acid tert-butyl ester. MS: m/z 635(M + H)⁺。

Example 156

Synthesis of 6- (4-aminopiperidin-1-yl) -2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide trihydrochloride

To {1- [6- (1H-indazol-6-ylcarbamoyl) -2- (2-trifluoromethyl-phenylamino) -3H-benzimidazol-5-yl]-piperidin-4-yl } -carbamic acid tert-butyl ester (0.25 mmol; see example 155) in methanol (1mL) was added 4M HCl in dioxane (0).5 mL). The resulting mixture was stirred at room temperature for 5-6 h. The volatiles were removed in vacuo and the resulting residue was suspended in diethyl ether. The resulting solid was collected by filtration, washed with diethyl ether and dried in vacuo to give 6- (4-amino-piperidin-1-yl) -2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide as the hydrochloride salt. MS: m/z 535(M + H)⁺。

Example 157

Synthesis of {1- [6- (1H-indazol-6-ylcarbamoyl) -2- (3-methylpyridin-2-ylamino) -3H-benzimidazol-5-yl ] -piperidin-4-yl } -carbamic acid tert-butyl ester

[1- [4, 5-diamino-2- (1H-indazol-6-ylcarbamoyl) -phenyl ] as described in general procedure B]Reaction of a solution of-piperidin-4-yl } -carbamic acid tert-butyl ester (0.3 mmol; see example 155) in DMF (1mL) with 1-isothiocyanato-2-trifluoromethylbenzene (0.3mmol) followed by in situ cyclization with EDC to give { {1- [6- (1H-indazol-6-ylcarbamoyl) -2- (3-methylpyridin-2-ylamino) -3H-benzimidazol-5-yl]-piperidin-4-yl } -carbamic acid tert-butyl ester. MS: m/z 582(M + H)⁺。

Example 158

Synthesis of 6- (4-aminopiperidin-1-yl) -2- (3-methyl-pyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide trihydrochloride

Using the procedure described in example 156, the product from example 157 was treated with 4M HCl in dioxane to give 6- (4-aminopiperidin-1-yl) -2- (3-methylpyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1)H-indazol-6-yl) -amide as the hydrochloride salt. MS: m/z 521(M + H)⁺。

Example 159

Synthesis of [5- (1H-indazol-6-ylethynyl) -1H-benzimidazol-2-yl ] - (2-trifluoromethyl-phenyl) -amine

A mixture of 4-bromo-2-nitrophenylamine (2.17g, 10mmol), ethynyltrimethyl-silane (2.11mL, 98%, 15mmol), dichlorobis (triphenylphosphine) palladium (II) (211mg, 0.3mmol) and copper (I) chloride (66.5mg, 0.35mmol) in THF (10mL) and triethylamine (10mL) was stirred at room temperature for 3 days. The product, 2-nitro-4-trimethylsilylethynyl phenylamine, was purified by silica gel column chromatography. LC-MS m/z: 235(M +1)⁺。

A mixture of the foregoing silyl intermediate, potassium carbonate (2.76g, 20mmol) and methanol (30mL) was stirred for two days. Purification by silica gel column chromatography gave 4-ethynyl-2-nitrophenylamine as a red solid (1.306g, 8.05mmol, 81% over 2 steps). LC-MS m/z: 163(M +1)⁺。

A mixture of 4-ethynyl-2-nitro-phenylamine (1.306g, 8.05mmol), 6-iodo-1H-indazole (1.965g, 8.05mmol), dichlorobis (triphenylphosphine) palladium (II) (122mg, 0.24mmol) and copper (I) chloride (54.4mg, 0.28mmol) in THF (8mL) and triethylamine (8mL) was stirred at room temperature overnight. Purification by silica gel column chromatography gave 4- (1H-indazol-6-ylethynyl) -2-nitrophenylamine as a red solid (777mg, 2.79mmol, yield 35%). LC-MSm/z: 279(M +1)⁺。

A mixture of the nitro compound (774mg, 2.78mmol), iron powder (1.61g, 97%, 28mmol) and ammonium chloride (2.25g, 42mmol) in ethanol (1.5mL) and water (1.5mL) was refluxed for 6 h. Purification by silica gel column chromatography gave 4- (1H-indazol-6-ylethynyl) -benzene-1, 2-diamine as a brown solid (284mg, 1.14mmol,yield 41%). LC-MS m/z: 249(M +1)⁺。

Reaction of the above diamine (0.3mmol) with 1-isothiocyanato-2-trifluoromethylbenzene (0.3mmol) followed by in situ cyclization with EDC gave [5- (1H-indazol-6-ylethynyl) -1H-benzimidazol-2-yl]- (2-trifluoromethylphenyl) -amine, as a yellow solid (178mg, 0.426mmol, 66% yield). LC-MS m/z: 418(M +1)⁺。

Example 160

Synthesis of 6-dimethylamino-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

To a solution of 4-amino-2-chloro-N- (1H-indazol-6-yl) -5-nitrobenzamide (1 mmol; see example 157) in DMF (1mL) was added 10% K₂CO₃Aqueous solution (0.25 mL). The mixture was then subjected to microwave irradiation at 80 ℃ for 60 min. The contents were cooled to RT and poured into ice cold water (20L). The resulting solid was collected by filtration, washed with water, and dried in vacuo to give 4-amino-2-dimethylamino-N- (1H-indazol-6-yl) -5-nitrobenzamide.

Reduction of the nitro compound (0.5mmol) under hydrogenation conditions as described in general procedure F gave 4, 5-diamino-2-dimethylamino-N- (1H-indazol-6-yl) benzamide.

Reaction of the above diamine (0.3mmol) with 1-isothiocyanato-2-trifluoromethylbenzene (0.3mmol) followed by in situ cyclization using EDC gave 6-dimethylamino-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide as described in general procedure B. MS: m/z 480(M + H)⁺。

Example 161

Synthesis of 6-dimethylamino-2- (3-methylpyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

4, 5-diamino-2-dimethylamino-N- (1H-indazol-6-yl) benzamide (see example 160; 0.3mmol) was reacted with 2-isothiocyanato-3-methylpyridine (0.3 mmol; prepared from 2-amino-3-methylpyridine and thiophosgene using the procedure described in example 115) followed by in situ cyclization with EDC to give 6-dimethylamino-2- (3-methylpyridin-2-ylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide as described in general procedure B. MS: m/z 427(M + H)⁺。

Example 162

Synthesis of 6- (4-methylpiperazin-1-yl) -2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid benzothiazol-5-ylamide

5-aminobenzothiazole (4.5mmol) was reacted with 4-chloro-2-fluoro-5-nitrobenzoyl chloride (5mmol) using the conditions described for example 133. The product, N-benzothiazol-5-yl-2-chloro-4-fluoro-5-nitrobenzamide, was also isolated in analogy to example 133.

Using the conditions described in example 115, a solution of the above amide (2mmol) in dioxane (4mL) was reacted with NH₄And (4) reacting an OH aqueous solution. After completion of the formation of 4-amino-N-benzothiazol-5-yl-2-chloro-5-nitrobenzamide, N-methylpiperazine (12mmol) was added to the reaction mixture. The contents were heated at reflux for 10h and the reaction mixture was cooled to RT. The contents were poured onto ice-cold water with vigorous stirring. The resulting solid was collected by filtration, washed with water and dried in vacuo to give the product 4-amino-N-benzothiazol-5-yl-2-, (4-methyl-piperazin-1-yl) -5-nitrobenzamide as a yellow solid.

The nitro compound (2mmol) obtained above was reduced under hydrogenation conditions as described in general procedure F to give 4, 5-diamino-N-benzothiazol-5-yl-2- (4-methylpiperazin-1-yl) -benzamide.

Reaction of the above diamine (0.3mmol) with 1-isothiocyanato-2-trifluoromethylbenzene (0.3mmol) followed by in situ cyclization using EDC gave 6- (4-methylpiperazin-1-yl) -2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid benzothiazol-5-ylamide as described in general procedure B. MS: m/z 552(M + H)⁺。

Example 163

4- [6- (benzothiazol-6-ylcarbamoyl) -2- (2-trifluoromethyl-phenylamino) -3H-benzimidazol-5-yl ] piperazine-1-carboxylic acid tert-butyl ester

To a solution of 4-amino-N-benzothiazol-6-yl-2-chloro-5-nitrobenzamide (2 mmol; prepared as in example 141) in dioxane (5mL) was added piperazine (10 mmol). The contents were heated at reflux for 10h and the reaction mixture was cooled to RT. The contents were poured onto ice-cold water with vigorous stirring. The resulting solid was collected by filtration, washed with water and dried in vacuo to give the product 4-amino-N-benzothiazol-6-yl-5-nitro-2-piperazin-1-yl-benzamide as a yellow solid.

The above amide (1mmol) was dissolved in THF (3mL), treated with BOC anhydride (1.2mmol) and stirred at RT for 2 h. The solvent was removed to dryness and the resulting residue was suspended in 10% EtOAc/hexanes (10mL) with stirring. The resulting solid was collected by filtration, washed with 10% EtOAc/hexanes, and dried in vacuo to give tert-butyl 4- [ 5-amino-2- (benzothiazol-6-ylcarbamoyl) -4-nitro-phenyl ] piperazine-1-carboxylate.

The nitro compound (0.8mmol) was reduced under hydrogenation conditions as described in general procedure F to give tert-butyl 4- [4, 5-diamino-2- (benzothiazol-6-ylcarbamoyl) -phenyl ] piperazine-1-carboxylate.

Reaction of the above diamine (0.3mmol) with 1-isothiocyanato-2-trifluoromethylbenzene (0.3mmol) followed by in situ cyclization with EDC gave 4- [6- (benzothiazol-6-ylcarbamoyl) -2- (2-trifluoromethylphenylamino) -3H-benzimidazol-5-yl]Piperazine-1-carboxylic acid tert-butyl ester. MS: m/z 638(M + H)⁺。

Example 164

Synthesis of 6-piperazin-1-yl-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide trihydrochloride

The product of example 163 was treated with 4M HCl in dioxane using the procedure described for example 156 to give 6-piperazin-1-yl-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide as the hydrochloride salt. MS: m/z 538(M + H)⁺。

Example 165

Synthesis of tert-butyl 4- [6- (benzothiazol-6-ylcarbamoyl) -2- (3-methylpyridin-2-ylamino) -3H-benzimidazol-5-yl ] piperazine-1-carboxylate

4- [4, 5-diamino-2- (benzothiazol-6-ylcarbamoyl) -phenyl ] as described in general procedure B]Piperazine-1-carboxylic acid tert-butyl ester (cf. example 163; 0.3mmol) with 2-isothiocyanato-3-methylpyridine(0.3 mmol; prepared from 2-amino-3-methylpyridine and thiophosgene using the procedure described in example 115) reaction followed by in situ cyclization using EDC gave 4- [6- (benzothiazol-6-ylcarbamoyl) -2- (3-methyl-pyridin-2-ylamino) -3H-benzimidazol-5-yl]Piperazine-1-carboxylic acid tert-butyl ester. MS: m/z 585(M + H)⁺。

Example 166

Synthesis of 2- (3-methyl-pyridin-2-ylamino) -6-piperazin-1-yl-1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide hydrochloride

The product of example 165 was treated with 4M HCl in dioxane using the procedure described for example 156 to give 2- (3-methyl-pyridin-2-ylamino) -6-piperazin-1-yl-1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide as the hydrochloride salt. MS: m/z 485(M + H)⁺。

Example 167

Synthesis of 2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide

To a solution of 3, 4-diaminobenzoic acid (3mmol) in DMF (10mL) was added 1-isothiocyanato-2-trifluoromethylbenzene (3.3mmol) and the resulting solution was stirred at RT for 4 h. After the thiourea formation was complete, solid K was added to the reaction mixture₂CO₃(10mmol) the mixture was heated at 90 ℃ for 10 h. The reaction mixture was cooled to RT and acidified to pH7 with 10% aqueous HCl. The contents were poured onto ice-cold water (30mL) with vigorous stirring. The resulting solid was collected by filtration, washed with water and dried in vacuo to give the product 2- (2-trifluoromethylbenzene)Phenylamino) -1H-benzimidazole-5-carboxylic acid as a yellow solid.

Using general procedure D, the carboxylic acid obtained above (0.25mmol) was coupled with 6-aminobenzothiazole (0.25mmol) using HBTU. After purification by silica gel chromatography using DCM/methanol as eluent, the product 2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid benzothiazol-6-ylamide was obtained as a light brown solid. MS: m/z 454(M + H)⁺。

Example 168

Synthesis of 6-piperazin-1-yl-2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (5-methyl-1H-indazol-6-yl) amide

To 2, 4-dimethylaniline (10mmol) in 5mL of concentrated H at 0 deg.C₂SO₄The mixture in (1) is added with fuming HNO dropwise₃(90%; 0.6 mL). The resulting mixture was stirred at RT for 12h and then poured slowly into ice. The solid was collected by filtration and dried to give 2, 4-dimethyl-5-nitroaniline as a yellow solid.

To a solution of nitroaniline (5mmol) obtained above in HOAc (5mL) was added dropwise isopentyl nitrile (6mmol) at RT. The resulting mixture was stirred at RT for 14h and then slowly poured onto cold saturated NaHCO₃Aqueous solution (15 mL). The contents were extracted with ethyl acetate (3X 20mL), the organic layers combined and washed with 5% Na₂CO₃Aqueous (30mL) wash. The volatiles were removed in vacuo to give 6-nitro-5-methylindazole as a brown solid.

The nitro compound (2mmol) obtained above was reduced under hydrogenation conditions as described in general procedure F to give 6-amino-5-methylindazole as a brown solid.

The aminoindazole (1.5mmol) described above was reacted with 4-chloro-2-fluoro-5-nitrobenzoyl chloride (1.5mmol) using the conditions described for example 115. The product 2-chloro-4-fluoro-N- (5-methyl-1H-indazol-6-yl) -5-nitrobenzamide was also isolated in analogy to example 115.

Using the conditions described in example 115, a solution of the above amide (1mmol) in dioxane (2mL) was reacted with NH₄And (4) reacting an OH aqueous solution. After completion of the formation of 4-amino-2-chloro-N- (5-methyl-1H-indazol-6-yl) -5-nitrobenzamide, piperazine (5mmol) was added. The contents were heated at reflux for 10h and the reaction mixture was cooled to RT. The contents were poured onto ice-cold water with vigorous stirring. The resulting solid was collected by filtration, washed with water and dried in vacuo to give the product 4-amino-N- (5-methyl-1H-indazol-6-yl) -5-nitro-2-piperazin-1-yl-benzamide as a yellow solid.

The above product (0.6mol) was treated with BOC anhydride using the procedure described in example 163.

Reduction of the nitroaniline (0.5mmol) under hydrogenation conditions as described in general procedure F gave tert-butyl 4- [4, 5-diamino-2- (5-methyl-1H-indazol-6-ylcarbamoyl) -phenyl ] piperazine-1-carboxylate.

Reaction of the above diamine (0.3mmol) with 1-isothiocyanato-2-trifluoromethylbenzene (0.3mmol) followed by in situ cyclization with EDC gave 4- [6- (5-methyl-1H-indazol-6-ylcarbamoyl) -2- (2-trifluoromethyl-phenylamino) -3H-benzimidazol-5-yl]Piperazine-1-carboxylic acid tert-butyl ester. MS: m/z 635(M + H)⁺。

The above product was treated with 4M HCl in dioxane using the procedure described for example 156 to give 6-piperazin-1-yl-2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (5-methyl-1H-indazol-6-yl) amide as the hydrochloride salt. MS: m/z 535(M + H)⁺。

Example 169

Synthesis of tert-butyl 4- [2- ((1S, 2S, 4R) -bicyclo [2.2.1] hept-2-ylamino) -6- (1H-indazol-6-ylcarbamoyl) -3H-benzimidazol-5-yl ] -piperazine-1-carboxylate

4-amino-2-chloro-N- (1H-indazol-6-yl) -5-nitrobenzamide (1 mmol; see example 154) was reacted with piperazine according to the procedure described for example 163 to give 4-amino-N- (1H-indazol-6-yl) -5-nitro-2-piperazin-1-ylbenzamide. The resulting product was treated with BOC anhydride as in example 163 to give 4- [ 5-amino-2- (1H-indazol-6-ylcarbamoyl) -4-nitro-phenyl ] -piperazine-1-carboxylic acid tert-butyl ester.

Reduction of the nitro compound (0.6mmol) under hydrogenation conditions as described in general procedure F gave tert-butyl 4- [4, 5-diamino-2- (1H-indazol-6-ylcarbamoyl) -phenyl ] piperazine-1-carboxylate.

Reaction of the above diamine (0.3mmol) with (S) -2-isothiocyanato-bicyclo [2.2.1] as described in general procedure B]Heptane (0.3mmol) followed by in situ cyclization using EDC gave 4- [2- ((1S, 2S, 4R) -bicyclo [2.2.1]Hept-2-ylamino) -6- (1H-indazol-6-ylcarbamoyl) -3H-benzimidazol-5-yl]-piperazine-1-carboxylic acid tert-butyl ester. MS: m/z 571(M + H)⁺。

Example 170

Synthesis of 2- ((1S, 2S, 4R) -bicyclo [2.2.1] hept-2-ylamino) -6-piperazin-1-yl-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) amide trihydrochloride salt

Using the procedure described for example 156, the product of example 169 was treated with 4M HCl in dioxane to give 2- ((1S, 2S, 4R) -bicyclo [2.2.1]Hept-2-ylamino) -6-piperazin-1-yl-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) amide as the hydrochloride salt. MS: m/z471(M + H)⁺。

Example 171

Synthesis of 6-chloro-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) amide

To a solution of 4-amino-2-chloro-N- (1H-indazol-6-yl) -5-nitrobenzamide (2 mmol; see example 154) in ethanol (5mL) and AcOH (1mL) was added iron powder (10 mmol). The reaction mixture was then heated to reflux for 6 h. The contents were cooled to RT, filtered through a pad of celite, and the pad was washed with ethanol. The filtrates were combined and concentrated in vacuo. The resulting residue was purified by column chromatography on silica gel using MeOH/DCM as eluent to give 4, 5-diamino-2-chloro-N- (1H-indazol-6-yl) benzamide.

Reaction of the above diamine (0.3mmol) with 1-isothiocyanato-2-trifluoromethylbenzene (0.3mmol) followed by in situ cyclization with EDC gave 6-chloro-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) amide as described in general procedure B. MS: m/z471(M + H)⁺。

Example 172

Synthesis of 2- ((1S, 2S, 4R) -bicyclo [2.2.1] hept-2-ylamino) -6-chloro-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) amide

4, 5-diamino-2-chloro-N- (1H-indazol-6-yl) benzamide (0.3 mmol; see example 171) was reacted with (1S, 2S, 4R) -2-isothiocyanato-bicyclo [2.2.1] as described in general procedure B]Heptane (0.3mmol) followed by in situ cyclization using EDC gave 2- ((1S, 2S, 4R)) -bicyclo [2.2.1]Hept-2-ylamino) -6-chloro-1H-benzimidazole-5-carboxylic acidAcid (1H-indazol-6-yl) amides. MS: m/z 421(M + H)⁺。

Example 173

Synthesis of 6- [4- (2-hydroxyethyl) -piperazin-1-yl ] -2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) amide

To a solution of 6-piperazin-1-yl-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide trihydrochloride (0.2 mmol; see example 88) in methanol (2mL) was added glyceraldehyde (2mmol) and the resulting mixture was stirred at RT for 60 min. Solid sodium borohydride (1mmol) was then added to the reaction mixture and stirring continued at RT for 10 h. The reaction mixture was then concentrated in vacuo and the residue was suspended in water (10mL) with vigorous stirring. After 30min, the solid was filtered, washed with water and dried under vacuum to give 6- [4- (2-hydroxyethyl) -piperazin-1-yl]-2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) amide as a white solid. MS: m/z 565(M + H)⁺。

Example 174

Synthesis of {4- [6- (1H-indazol-6-ylcarbamoyl) -2- (2-trifluoromethyl-phenylamino) -3H-benzimidazol-5-yl ] -piperazin-1-yl } acetic acid

To a solution of 6-piperazin-1-yl-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide trihydrochloride (0.25 mmol; see example 88) in methanol (1mL) was added glyoxylic acid (0.5mmol) and the resulting mixture was stirred at RT for 60 min. Solid sodium borohydride (0.6mmol) was then added to the reaction mixture, followed byStirring was continued at RT for 10 h. A few drops of glacial acetic acid were then added to the reaction mixture and the mixture was stirred for 30 min. The volatiles were then removed in vacuo and the residue was suspended in water (10mL) with vigorous stirring. After 30min, the solid was filtered, washed with water and dried under vacuum to give {4- [6- (1H-indazol-6-ylcarbamoyl) -2- (2-trifluoromethyl-phenylamino) -3H-benzimidazol-5-yl]-piperazin-1-yl } acetic acid as a white solid. MS: m/z 579(M + H)⁺。

Example 175

Synthesis of 6- (4-dimethylsulfamoyl-piperazin-1-yl) -2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

To a solution of 6-piperazin-1-yl-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide trihydrochloride (0.3 mmol; see example 88) in DMF (1mL) was added triethylamine (1.5mmol) and N, N-dimethylsulfamoyl chloride (0.4 mmol). The resulting mixture was stirred at RT for 4h and hydrazine hydrate (2mmol) was added. The contents were warmed to 50 ℃ and stirred vigorously for 60 min. The reaction mixture was then poured into ice-cold water, the solid filtered, washed with water and dried under vacuum. The crude product was then purified by silica gel column chromatography using MeOH/DCM as eluent to give 6- (4-dimethylsulfamoyl-piperazin-1-yl) -2- (2-trifluoromethyl-phenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide as a white solid. MS: m/z 628(M + H)⁺。

Example 176

Synthesis of {6- [5- (1H-indazol-6-yl) -1H-imidazol-2-yl ] -1H-benzimidazol-2-yl } - (2-trifluoromethylphenyl) -amine

Methyl-3-nitroacetophenone (10mmol) was reduced under hydrogenation conditions as described in general procedure F to give 1- (3-amino-4-methyl-phenyl) ethanone (1.4 g).

Concentrated HCl (2mL) was added to 1- (3-amino-4-methyl-phenyl) ethanone (8.4mmol) and NaBF₄(1.2g, 11mmol) in H₂In a mixture in O (10mL), the solution was cooled to 0 ℃. NaNO is dripped₂(0.58g, 8.4mmol) of H₂O (1.5mL) solution, and the mixture was stirred at 0 ℃ for 30 min. The resulting solid was collected by filtration and washed with H₂O (5mL) followed by Et₂O (5mL) and dried under reduced pressure. Adding CH to the solid₂Cl₂(20mL), KOAc (0.91g, 9.3mmol) and 18-crown-6 (50mg, 0.2mmol), and the mixture was stirred at room temperature for 4 h. Addition of H₂O (20mL), the layers were separated. The organic layer was dried (MgSO₄) The solvent was removed under reduced pressure to give 1- (1H-indazol-6-yl) ethanone (0.52 g).

Pyrrolidone trihydrobromide (1.8g, 3.6mmol) was added to a solution of 1- (1H-indazol-6-yl) ethanone (0.5g, 3mmol) in THF (10mL) and the solution was heated at reflux for 2H. The solution was allowed to cool to room temperature and H was added₂O (30mL), the mixture was extracted with EtOAc (3X 20mL), dried (MgSO)₄). The solvent was removed under reduced pressure to give 2-bromo-1- (1H-indazol-6-yl) -ethanone, which was used in the next step without purification.

DIEA (0.7mL, 3.6mmol) was added to a solution of 2-bromo-1- (1H-indazol-6-yl) ethanone (3mmol) and 4-amino-3-nitrobenzoic acid (0.643g, 3.5mmol) in DMF (10mL) and the solution was stirred at room temperature for 2H. Adding NH to the solution₄OAc (5g, 65mmol) followed by HOAc (10mL), the mixture was stirred at 140 ℃ for 2 h. The mixture was cooled to room temperature and poured into H₂O (30 mL). Filtering to collect precipitate, and purifying with H₂O (10mL) and dried under reduced pressure to give 4- [5- (1H-indazol-6-yl) -1H-imidazol-2-yl]2-Nitrophenylamine (0.56 g).

Reduction of the nitroaniline (1mmol) under hydrogenation conditions as described in general procedure F gave 4- [5- (1H-indazol-6-yl) -1H-imidazol-2-yl ] -benzene-1, 2-diamine.

Reaction of the above diamine (0.5mmol) with 1-isothiocyanato-2-trifluoromethylbenzene (0.5mmol) followed by in situ cyclization with EDC gave {6- [5- (1H-indazol-6-yl) -1H-imidazol-2-yl]-1H-benzimidazol-2-yl } - (2-trifluoromethylphenyl) -amine. MS: m/z 460(M + H)⁺。

Example 177

Synthesis of 6- (2-dimethylamino-ethylsulfanyl) -2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

NaH (2mmol) was added to a solution of 2-dimethylaminoethanethiol (2mmol) in NMP (2mL) and the mixture was stirred at room temperature for 10 min. 4-amino-2-chloro-N- (1H-indazol-6-yl) -5-nitrobenzamide (1 mmol; see example 154) was added to the mixture and the mixture was stirred at 60-65 ℃ for 3H. Water (4mL) was added to the mixture, and the mixture was extracted with EtOAc (3X 10mL) over MgSO₄And (5) drying. The combined extracts were dried (MgSO)₄) The solvent was removed under reduced pressure to give the desired product 4-amino-2- (2-dimethylaminoethylthio) -N- (1H-indazol-6-yl) -5-nitro-benzamide without further purification.

Reduction of the nitroaniline (1mmol) under hydrogenation conditions as described in general procedure F gave 4, 5-diamino-2- (2-dimethylaminoethylthio) -N- (1H-indazol-6-yl) -benzamide.

Reaction of the above diamine (0.5mmol) with 1-isothiocyanato-2-trifluoromethylbenzene (0.5mmol) followed by in situ cyclization with EDC gave 6- (2-dimethylaminoethylthio) -2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide as described in general procedure B。MS：m/z 540(M+H)⁺。

Example 178

Synthesis of 5-ethyl-8- (1H-indazol-6-yl) -2- (2-trifluoromethylphenylamino) -5, 6, 7, 8-tetrahydro-3H-1, 3, 5, 8-tetraazacyclohepta [ f ] inden-9-one

To a solution of 4-amino-2-chloro-N- (1H-indazol-6-yl) -5-nitrobenzamide (5 mmol; see example 154) in dioxane (10mL) was added 2-ethylaminoethanol (15 mmol). The resulting mixture was heated at reflux for 10 h. The reaction mixture was cooled to room temperature and diluted with water (20 mL). The resulting solid was collected by filtration, washed with water and dried in vacuo. The product 4-amino-2- [ ethyl (2-hydroxyethyl) amino ] -N- (1H-indazol-6-yl) -5-nitrobenzamide was used without any purification.

Adding MeSO₂Cl (0.5mL, 6.3mmol) was added dropwise to a solution of DIEA (1.6mL) and pyridine (1.5mL) in the above nitroaniline (1g, 3.0mmol) in THF (10 mL). The solution was stirred at room temperature for 1h and poured into water (10 mL). The mixture was extracted with EtOAc (3X 10mL), and the combined extracts were MgSO₄And (5) drying. The solvent was removed under reduced pressure to give methanesulfonic acid 2- { [ 5-amino-2- (1-methanesulfonyl-1H-indazol-6-ylcarbamoyl) -4-nitrophenyl]Ethylamino } ethyl ester (1.4g, 2.6 mmol).

NaH (60%, 266mg, 6.7mol) was added to crude methanesulfonic acid 2- { [ 5-amino-2- (1-methanesulfonyl-1H-indazol-6-ylcarbamoyl) -4-nitrophenyl]Ethylamino } ethyl ester (2.6mmol) in THF (10 mL). The solution was stirred at reflux for 3 h. The solvent was removed under reduced pressure and the residue was dissolved in EtOAc and washed with water (10 mL). The organic layer was separated over MgSO₄Drying and removal of the solvent under reduced pressure to give 8-amino-1-ethyl-4- (1-methanesulfonyl-1H-indazol-6-yl) -7-nitro-1, 2, 3, 4-tetrahydro-benzeneAnd [ e ]][1，4]Diaza derivatives-5-ketone (1.1g, 2.5 mmol).

Hydrazine (0.6mL) was added to 8-amino-1-ethyl-4- (1-methanesulfonyl-1H-indazol-6-yl) -7-nitro-1, 2, 3, 4-tetrahydro-benzo [ e ]][1，4]Diaza derivatives-5-ketone (1.1g, 2.5mmol) in 1: 1THF/MeOH (20 mL). The solution was stirred at room temperature for 16 h. The solvent was removed under reduced pressure to give 8-amino-1-ethyl-4- (1H-indazol-6-yl) -7-nitro-1, 2, 3, 4-tetrahydrobenzo [ e ]][1，4]Diaza derivatives-5-one (815 mg).

Reduction of the nitroaniline (1mmol) under hydrogenation conditions as described in general procedure F gave 7, 8-diamino-1-ethyl-4- (1H-indazol-6-yl) -1, 2, 3, 4-tetrahydrobenzo [ e ]][1，4]Diaza derivatives-5-ketones.

Reaction of the above diamine (0.5mmol) with 1-isothiocyanato-2-trifluoromethylbenzene (0.5mmol) followed by in situ cyclization with EDC gave 5-ethyl-8- (1H-indazol-6-yl) -2- (2-trifluoromethylphenylamino) -5, 6, 7, 8-tetrahydro-3H-1, 3, 5, 8-tetraazacyclohepta [ f ] triene as described in general procedure B]Inden-9-one. MS: m/z 506(M + H)⁺。

Example 179

Synthesis of 6-imidazol-1-yl-2- (2-trifluoromethylphenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

To a solution of 4-amino-2-chloro-N- (1H-indazol-6-yl) -5-nitrobenzamide (1 mmol; see example 154) in NMP (2mL) was added imidazole (5 mmol). The resulting mixture was subjected to microwave irradiation at 120 ℃ for 2 h. The reaction mixture was cooled to room temperature and diluted with water (30 mL). The resulting solid was collected by filtration, washed with water and dried in vacuo. The product, 4-amino-2-imidazol-1-yl-N- (1H-indazol-6-yl) -5-nitro-benzamide, was obtained as a yellow solid and used without any purification.

The nitro compound (0.5mmol) obtained above was reduced under hydrogenation conditions as described in general procedure F to give 4, 5-diamino-2-imidazol-1-yl-N- (1H-indazol-6-yl) -benzamide.

Reaction of the above diamine (0.3mmol) with 1-isothiocyanato-2-trifluoromethylbenzene (0.3mmol) followed by in situ cyclization using EDC gave 6-imidazol-1-yl-2- (2-trifluoromethylphenylamino) -3H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide as described in general procedure B. MS: m/z 503(M + H)⁺。

Example 180

Synthesis of 2- (2-trifluoromethylphenylamino) -benzoxazole-5-carboxylic acid (1H-indazol-6-yl) -amide

Following general procedure E, using 4-hydroxy-3-nitrobenzoic acid (5mmol) and 6-aminoindazole (5mmol), 4-hydroxy-N- (1H-indazol-6-yl) -3-nitrobenzamide was prepared as a yellow solid.

Reduction of the nitrophenol obtained above (3mmol) under hydrogenation conditions as described in general procedure F gave 3-amino-4-hydroxy-N- (1H-indazol-6-yl) -benzamide.

To a solution of the above aminophenol (0.5mmol) in DMF (2mL) was added 1-isothiocyanato-2-trifluoromethylbenzene (0.6mmol) and DIEA (1 mmol). Make contraryThe mixture was subjected to microwave irradiation at 120 ℃ for 1 h. The reaction mixture was cooled to room temperature and diluted with water (20 mL). The resulting solid was collected by filtration, washed with water and dried in vacuo. The crude product was purified by column chromatography on silica gel using MeOH/DCM as eluent to give 2- (2-trifluoromethylphenylamino) -benzooxazole-5-carboxylic acid (1H-indazol-6-yl) -amide as a yellow solid. MS: m/z 438(M + H)⁺。

Example 181

Synthesis of 2- (1-benzyl-1H-imidazol-2-ylamino) -6- (4-methylpiperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

A solution of 2-chloro-4-fluoro-N- (1H-indazol-6-yl) -5-nitrobenzamide (10mmol) in dioxane (20mL) was reacted with NH using the conditions described in example 115₄And (4) reacting an OH aqueous solution. After completion of the formation of 2-amino-4-fluoro-N- (1H-indazol-6-yl) -5-nitrobenzamide, N-methylpiperazine (40mmol) (NMP) was added to the reaction mixture. The contents were heated at reflux for 10h and the reaction mixture was cooled to RT. The contents were poured onto ice-cold water with vigorous stirring. The resulting solid was collected by filtration, washed with water and dried in vacuo to give the product 4-amino-N- (1H-indazol-6-yl) -2- (4-methylpiperazin-1-yl) -5-nitrobenzamide as a yellow solid.

The nitro compound (6mmol) obtained above was reduced under hydrogenation conditions as described in general procedure F to give 4, 5-diamino-2- (4-methylpiperazin-1-yl) -N- (1H-indazol-6-yl) -benzamide.

Benzyl bromide (3mmol) and K₂CO₃(6mmol) was added to a solution of 2-nitroimidazole (2mmol) in DMF (6 mL). The mixture was stirred at 60-70 ℃ for 4h or overnight. The contents were cooled to room temperature and water (30mL) was added. The mixture was extracted with EtOAc (3X 15 mL). Combining the extracts over MgSO₄Drying, filtering and removing the solvent in vacuo to obtain 1-benzyl-2-nitro-1H-imidazole. The product was used for further transformation without further purification.

The procedure described in example 159 was used to reduce the nitroimidazole described above (1.5mmol) using iron powder and ammonium chloride to give 1-benzyl-2-amino-1H-imidazole, which was used without any purification.

Following general procedure A, the above aminoimidazole was converted to 1-benzyl-2-isothiocyanato-1H-imidazole.

Coupling of isothiocyanate (1mmol) with 3, 4-diamino-N- (1H-indazol-6-yl) -benzamide (1mmol) followed by cyclization with EDC gave 2- (1-benzyl-1H-imidazol-2-ylamino) -6- (4-methylpiperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide as described in general procedure B. MS: m/z 547(M + H)⁺。

Example 182

Synthesis of 4- [2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6- (1H-indazol-6-ylcarbamoyl) -3H-benzimidazol-5-yl ] -piperazine-1-carboxylic acid tert-butyl ester

1-cyclopentyl-2-nitro-1H-imidazole was prepared according to the alkylation procedure described in example 181 using bromocyclopentane (14mmol) and 2-nitroimidazole (10 mmol). The resulting product was reduced under hydrogenation conditions as described in general procedure F to give 1-cyclopentyl-2-amino-1H-imidazole. This aminoimidazole derivative was converted into 1-cyclopentyl-2-isothiocyanato-1H-imidazole according to general procedure a.

Isothiocyanate (1mmol) was reacted with 4- [4, 5-diamino-2- (1H-indazol-6-ylcarbamoyl) -phenyl as described in general procedure B]Coupling of piperazine-1-carboxylic acid tert-butyl ester (1 mmol; cf. example 169) followed by cyclization using EDC gave 4- [2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6- (1H-indazol-6-ylcarbamoyl) -3H-benzimidazol-5-yl]-piperazine-1-carboxylic acid tert-butyl ester. MS: m/z 611(M + H)⁺。

Example 183

Synthesis of 2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6-piperazin-1-yl-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) amide trihydrochloride

The product of example 182 was treated with 4M HCl in dioxane using the procedure described for example 156 to give 2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6-piperazin-1-yl-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) amide as the hydrochloride salt. MS: m/z 511(M + H)⁺。

Example 184

Synthesis of 2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6- (4-isopropylpiperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) amide

To a solution of 4-amino-2-chloro-N- (1H-indazol-6-yl) -5-nitrobenzamide (1 mmol; see example 154) in dioxane (2mL) was added N-isopropylpiperazine (4 mmol). The resulting mixture was heated at reflux for 10 h. The reaction mixture was cooled to room temperature, diluted with water (20mL) while stirring vigorously. The resulting solid was collected by filtration, washed with water and dried in vacuo to give the product 4-amino-N- (1H-indazol-6-yl) -2- (4-isopropyl-piperazin-1-yl) -5-nitro-benzamide as a yellow solid.

Reduction of the nitro compound (0.5mmol) under hydrogenation conditions as described in general procedure F gave 4, 5-diamino-N- (1H-indazol-6-yl) -2- (4-isopropylpiperazin-1-yl) -benzamide.

Coupling of the above diamine (0.3mmol) with 1-cyclopentyl-2-isothiocyanato-1H-imidazole (0.3 mmol; see example 182) followed by in situ cyclization with EDC gave 2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6- (4-isopropylpiperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) amide as described in general procedure B. MS: m/z 553(M + H)⁺。

The compounds listed in Table 13 were synthesized according to the procedure for example 184 using 4-amino-2-chloro-N- (1H-indazol-6-yl) -5-nitrobenzamide.

Watch 13

Examples	R	MS(m/z)
			185	4-ethylpiperazin-1-yl	539
186	(2-dimethylaminoethyl) -methylamino	527
			187	4-methyl [1, 4 ]]Diazepan-1-yl	539

Using general procedure B, the following compounds (table 14) were synthesized using 1-alkyl-2-isothiocyanato-1H-imidazole (prepared using the procedure of example 182) and 4, 5-diamino-2- (4-methylpiperazin-1-yl) -N- (1H-indazol-6-yl) -benzamide (see example 181):

TABLE 14

Examples	R	MS(m/z)
			188	Cyclohexyl radical	539
189	Methyl radical	471
			190	Cyclohexyl methyl radical	553
191	Isobutyl radical	513
			192	Cyclobutyl radical	511
193	1-Ethyl propyl	527
			194	N-butyl	513
195	2-methoxyethyl group	515
			196	Ethyl radical	485

Using general procedure B, the following compounds (table 15) were synthesized using 1-alkyl-2-isothiocyanato-1H-imidazole (prepared using the procedure of example 182) and 4, 5-diamino-N-benzothiazol-6-yl-2- (4-methylpiperazin-1-yl) -benzamide (see example 141):

watch 15

Examples	R	MS(m/z)
			197	2-methoxyethyl group	532
198	Ethyl radical	502

Example 199

Synthesis of 2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6- (4-methylpiperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-5-yl) -amide

Using the procedure of example 115, 5-aminobenzothiazole (5mmol) was reacted with 4-chloro-2-fluoro-5-nitrobenzoyl chloride (5 mmol). The product, 2-chloro-4-fluoro-N- (1H-indazol-5-yl) -5-nitrobenzamide, was used for further conversion without any purification.

A solution of 2-chloro-4-fluoro-N- (1H-indazol-5-yl) -5-nitrobenzamide (2mmol) in dioxane (4mL) was reacted with NH using the conditions described in example 115₄OH waterAnd (4) reacting the solution. After completion of the formation of 2-amino-4-fluoro-N- (1H-indazol-6-yl) -5-nitrobenzamide, N-methylpiperazine (8mmol) was added to the reaction mixture. The contents were heated at reflux for 10h and the reaction mixture was cooled to RT. The contents were poured onto ice-cold water with vigorous stirring. The resulting solid was collected by filtration, washed with water and dried in vacuo to give the product 4-amino-N- (1H-indazol-5-yl) -2- (4-methylpiperazin-1-yl) -5-nitrobenzamide as a yellow solid.

The nitro compound (1mmol) obtained above was reduced under hydrogenation conditions as described in general procedure F to give 4, 5-diamino-2- (4-methylpiperazin-1-yl) -N- (1H-indazol-5-yl) -benzamide.

Coupling of the above diamine (0.3mmol) with 1-cyclopentyl-2-isothiocyanato-1H-imidazole (0.3 mmol; see example 182) followed by in situ cyclization with EDC gave 2- (1-cyclopentyl-1H-imidazol-2-ylamino) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-5-yl) -amide as described in general procedure B. MS: m/z 525(M + H)⁺。

The compounds in table 16 below were synthesized using the procedure described in example 199.

TABLE 16

Examples	Ar	MS(m/z)
			200	1H-benzotriazol-5-yl	526
201	Benzothiazol-6-yl	542
			202	2-oxo-2, 3-dihydro-1H-indol-5-yl	540
203	1H-indol-6-yl	524
			204	3H-benzimidazol-5-yl	525
205	Benzothiazol-5-yl	542

Example 206

Synthesis of 2- (1-thietane-3-yl-1H-imidazol-2-ylamino) -6- (4-methylpiperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-5-yl) -amide

2-Nitro-imidazole (0.5g, 4.4mmol) was addedTo a solution of KOH (6.6mmol) in water (10 mL). To this solution was added 2- (chloromethyl) thiacyclopropane (0.72g, 6.6mmol) and the solution was stirred at 65-70 ℃ for 1 h. Distilling off the solvent, purifying the residue by flash column chromatography with CH₂Cl₂As eluent, 0.43g of the desired product 2-nitro-1-thien-3-yl-1H-imidazole (52%) is obtained.

The nitro compound (2mmol) obtained above was reduced under hydrogenation conditions as described in general procedure F to give 2-amino-1-thietane-3-yl-1H-imidazole.

Following general procedure A, the above aminoimidazole derivatives were converted to 1-thietane-3-yl-2-isothiocyanato-1H-imidazole.

Coupling of 4, 5-diamino-2- (4-methylpiperazin-1-yl) -N- (1H-indazol-6-yl) -benzamide (0.5 mmol; see example 181) with 1-thietane-3-yl-2-isothiocyanato-1H-imidazole (0.5mmol) followed by in situ cyclization with EDC gave 2- (1-thietane-3-yl-1H-imidazol-2-ylamino) -6- (4-methylpiperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-5-yl) -amide as described in general procedure B. MS: m/z 529(M + H)⁺。

Example 207

Synthesis of 2-amino-6- (4-methylpiperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide hydrobromide

To a solution of 4, 5-diamino-2- (4-methylpiperazin-1-yl) -N- (1H-indazol-6-yl) -benzamide (2 mmol; see example 181) in 10% aqueous EtOH (6mL) was added cyanogen bromide (2.2mmol) and the mixture was heated at reflux for 4H. The reaction mixture was then concentrated in vacuo and the resulting residue was suspended in diethyl ether with vigorous stirring. The resulting solid was collected by filtration, washed with diethyl ether, and dried in vacuo to give 2-amino-6- (4)-methylpiperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide as the hydrobromide salt. MS: m/z391(M + H)⁺。

Example 208

Synthesis of 2- (3-cyclopentyl-3-ethylureido) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

A solution of cyclopentylethylamine (3mmol) in dry THF (3mL) was added dropwise to phosgene (4mmol) at 0 deg.C. After the addition was complete, the reaction mixture was stirred at 0 ℃ for 30 min. The volatiles were removed in vacuo and the resulting residue was dried in vacuo. The crude product cyclopentylethylcarbamoyl chloride was used for further conversion without any purification.

To a solution of 2-amino-6- (4-methylpiperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide hydrobromide (0.5mmol) in DMF (2mL) at RT was added DIEA (2mmol) followed by carbamoyl chloride (0.6mmol) obtained above. The resulting mixture was stirred for 4 h. Hydrazine hydrate (0.25mL) was added to the reaction mixture. The contents were warmed to 50 ℃ and stirred for 60 min. The reaction mixture was then cooled to RT, diluted with ice cold water (10mL) and extracted with EtOAc (2 × 10 mL). The combined extracts were washed with water (10mL) and brine (10 mL). After removal of the solvent, the resulting residue was purified by silica gel column chromatography to give 2- (3-cyclopentyl-3-ethylureido) -6- (4-methyl-piperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide. MS: m/z 530(M + H)⁺。

Example 209

Synthesis of 2-mercapto-6- (4-methylpiperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

To a solution of 4, 5-diamino-2- (4-methylpiperazin-1-yl) -N- (1H-indazol-6-yl) -benzamide (0.5 mmol; see example 181) in DMF (1mL) was added thiocarbonyldiimidazole (0.55 mmol). After the addition, the mixture was heated at 45 ℃ for 1 h. The reaction mixture was cooled to RT and the contents poured onto ice-cold water with vigorous stirring. The resulting solid was collected by filtration, washed with water and dried in vacuo to give the product 2-mercapto-6- (4-methylpiperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide as a yellow solid.

Example 210

Synthesis of 2- (1-cyclopentyl-1H-benzimidazol-2-ylamino) -6- (4-methylpiperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide

To a solution of 2-fluoro-1-nitrobenzene (2mmol) in THF were added cyclopentylamine (2.5mmol) and K₂CO₃(3 mmol). The resulting mixture was heated at 60 ℃ for 4 h. The contents were cooled to RT and the solid was filtered off. The filtrate was concentrated in vacuo and the residue was dried in vacuo. The product 2-cyclopentylamino-1-nitrobenzene was used for further conversion without any purification.

Nitroaniline (2mmol) obtained above was reduced under hydrogenation conditions as described in general procedure F to give N-cyclopentylbenzene-1, 2-diamine.

To a solution of the diamine (1.5mmol) obtained above in 10% aqueous EtOH (4mL) was added cyanogen bromide (1.7mmol) and heated at reflux for 4 h. The reaction mixture was then cooled to RT and solid K was added₂CO₃(2mmol), stirred vigorously for 30 min. The solid was then filtered off and the filtrate was concentrated in vacuo to give 1-cyclopentyl-1H-benzimidazol-2-ylamine, which was used without any purification for further work-upAnd (4) transformation.

Following general procedure A, the above aminobenzimidazole (1mmol) derivative was converted to 1-cyclopentyl-2-isothiocyanato-1H-benzimidazole.

4, 5-diamino-2- (4-methylpiperazin-1-yl) -N- (1H-indazol-6-yl) -benzamide (0.5 mmol; see example 181) was coupled with 1-cyclopentyl-2-isothiocyanato-1H-benzimidazole (0.5mmol) followed by in situ cyclization with EDC to give 2- (1-cyclopentyl-1H-benzimidazol-2-ylamino) -6- (4-methylpiperazin-1-yl) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide as described in general procedure B. MS: m/z 575(M + H)⁺。

Example 211

Synthesis of [6- (1H-indazol-6-yloxy) -1H-benzimidazol-2-yl ] - (2-trifluoromethylphenyl) -amine

To a stirred suspension of 6-aminoindazole (20mmol) in concentrated HCl (6mL) at 0 deg.C, NaNO was added in portions₂(22mmol) in water (12 mL). During the addition, the temperature of the reaction mixture was maintained at 0-5 ℃ and stirring was continued for another 45 min. The contents were then added to a flask containing 1% aqueous HCl (200mL) and heated at 100 ℃. The reaction mixture was then stirred at 100 ℃ for 5 h. The contents were cooled to RT and over 5% Na₂CO₃The aqueous solution was neutralized to pH7 and extracted with EtOAc (2X 70 mL). The combined organic layers were washed with brine and anhydrous Na₂SO₄And (5) drying. The solvent was removed under vacuum to give the 6-hydroxyindazole as a dark brown solid, which was used for further conversion without any purification.

To a stirred solution of 2-chloro-4-fluoro-1-nitrobenzene (3mmol) in DMF (5mL) was added 6-hydroxyindazole (3mmol) and K₂CO₃(6 mmol). The contents were heated at 90 ℃ for 6 h. The reaction mixture was cooled to RT and the contents wereThe contents were poured onto ice-cold water with vigorous stirring. The resulting solid was collected by filtration, washed with water and dried in vacuo to give the product 6- (3-chloro-4-nitrophenoxy) -1H-indazole as a yellow solid which was used for further conversion without any purification.

To a stirred solution of the nitro compound (2mmol) in DMF (4mL) was added benzylamine (4mmol) and the contents heated at 100 ℃ for 6 h. The reaction mixture was cooled to RT and the contents poured onto ice-cold water with vigorous stirring. The resulting solid was collected by filtration, washed with water and dried in vacuo. The resulting residue was purified by column chromatography on silica gel using hexane/EtOAC as eluent to give the product benzyl- [5- (1H-indazol-6-yloxy) -2-nitrophenyl ] -amine as a yellow solid.

Reduction of nitroaniline (1mmol) obtained above under hydrogenation conditions as described in general procedure F gave 4- (1H-indazol-6-yloxy) -benzene-1, 2-diamine.

Coupling of the above diamine (0.3mmol) with 1-isothiocyanato-2-trifluoromethylbenzene (0.3mmol) followed by in situ cyclization with EDC gave [6- (1H-indazol-6-yloxy) -1H-benzimidazol-2-yl]- (2-trifluoromethylphenyl) -amine. MS: m/z 410(M + H)⁺。

Example 212

Synthesis of {5- [2- (1H-indazol-6-yl) -ethyl ] -1H-benzimidazol-2-yl } - (2-trifluoromethylphenyl) amine

Reduction of [5- (1H-indazol-6-ylethynyl) -1H-benzimidazol-2-yl under 50psi hydrogenation conditions as described in general procedures]- (2-trifluoromethylphenyl) -amine (0.3 mmol; see example 159) in ethanol (3mL) to give {5- [2- (1H-indazol-6-yl) -ethyl]-1H-benzimidazol-2-yl } - (2-trifluoromethylphenyl) amine. MS: m/z 422(M + H)⁺。

Example 213

Synthesis of 3- [ 6-diethylamino-5- (1H-indazol-6-ylcarbamoyl) -1H-benzimidazol-2-ylamino ] -benzoic acid

To 3- [ 6-diethylamino-5- (1H-indazol-6-ylcarbamoyl) -1H-benzimidazol-2-ylamino]Methyl benzoate (0.2 mmol; see example 115) in methanol (2mL) and THF (2mL) was added 1M aqueous LiOH (2 mL). The resulting solution was then stirred at RT until the reaction was complete. The pH of the reaction mixture was adjusted to pH4-5 with 5% aqueous citric acid. The resulting solid was filtered, washed with ice-cold water and dried under vacuum to give 3- [ 6-diethylamino-5- (1H-indazol-6-ylcarbamoyl) -1H-benzimidazol-2-ylamino]-benzoic acid as a white solid. MS: m/z 484(M + H)⁺。

Example 214

Synthesis of 3- [5- (benzothiazol-6-ylcarbamoyl) -6-diethylamino-1H-benzimidazol-2-ylamino ] -benzoic acid methyl ester

Coupling of 4, 5-diamino-N-benzothiazol-6-yl-2-diethylaminobenzamide (0.3 mmol; see example 133) with methyl 3-isothiocyanatobenzoate (0.3mmol) followed by in situ cyclization with EDC gave 3- [5- (benzothiazol-6-ylcarbamoyl) -6-diethylamino-1H-benzimidazol-2-ylamino]-benzoic acid methyl ester. MS: m/z 515(M + H)⁺。

Example 215

Synthesis of 3- [5- (benzothiazol-6-ylcarbamoyl) -6-diethylamino-1H-benzimidazol-2-ylamino ] -benzoic acid

Hydrolysis of the methyl ester from example 214 (0.2mmol) using the conditions described in example 216 gave 3- [5- (benzothiazol-6-ylcarbamoyl) -6-diethylamino-1H-benzimidazol-2-ylamino]-benzoic acid as a white solid. MS: m/z 501(M + H)⁺。

Biological data

The compounds of the present invention elicit a measurable pharmacological response. The compounds of the invention in Table 1 have Aurora kinase binding affinity (IC)₅₀< 1 μ M), may be Aurora kinase selective compared to other kinases. In addition to binding to Aurora kinase, the compounds of the present invention may also measurably inhibit the proliferation of tumor cells.

Example 216

Aurora A, B, C enzyme assay

The Aurora kinase assay utilizes the peptide substrate biotin-ahx-LRRWSLGLRRWSLG as the phosphoryl acceptor.

Assays were performed in 96-well U-bottom plates. Aurora A and Aurora C enzymes were purchased from PanVera, and Aurora B enzymes were purchased from BPS Bioscience. Compounds were diluted in DMSO prior to addition in the assay. In general, the assay is performed by incubating the enzyme (0.2-10nM) with or without inhibitor, 0.1-1. mu. Ci. gamma. at 37 deg.C³³P-ATP(PerkinElmer)、0.1-100μM ATP、0.1-10mM MnCl₂1-10. mu.M sodium orthovanadate, 1-10mM DTT and 1-100. mu.M peptide for 5-120min, the final assay volume being 60. mu.L. The buffer used to make up the final assay volume to 60. mu.L was 50mM MOPS, pH7.0, containing 1-5% DMSO and 0.05% BSA. The reaction was terminated by the addition of 0.2-2 volumes of 0.75% phosphoric acid.

Detection of peptide phosphorylation was accomplished as follows, and after peptides were collected on P8196 well filter plates (Whatman), scintillation counting was performed using a beta counter (TopCount). Total control cpm (C)⁺) Cpm (C) against background^-) Wells contained DMSO instead of compound. Background control (C)^-) The pores lack peptides. Assume total number (C)⁺) Subtract background (C)^-) The number is proportional to the initial reaction rate. The percent enzyme inhibition was calculated as 1- [ (cpm)_{Sample (I)}-C^-)/(C⁺-C^-)]X 100%. According to the four parameter logic equation Y ═ bottom + (top-bottom)/1 +10^ ((LogEC50-X)^＊Hill slope)), where X is the logarithm of the compound concentration and Y is the percent inhibition, using GraphPadPrism^TMDetermination of IC from enzyme inhibition-Compound concentration Curve₅₀The value is obtained.

In the above assay, each compound in Table 1 exhibits an IC less than or equal to 1.0 μ M for at least one Aurora kinase A, B or C₅₀The value is obtained.

Example 217

EGF RTK enzyme assay

EGF receptor tyrosine kinase assay utilizes the peptide substrate biotin-ahx-EEEEYFELVAKKK-C (O) NH₂(Advanced Chemtech, # PX9197) as a phosphoryl acceptor.

Assays were performed in 96-well U-bottom plates. The tyrosine kinase domain of EGF receptor was purchased from Upstate (# 14-531). Compounds were diluted in DMSO prior to addition in the assay. In general, the assay is performed by incubating the enzyme (0.2-10nM) with or without inhibitor, 0.1-1. mu. Ci. gamma. at 37 deg.C³³P-ATP(Perkin Elmer)、0.1-100μMATP、0.1-10mM MnCl₂1-10. mu.M sodium orthovanadate, 1-10mM DTT and 1-100. mu.M peptide for 5-120min, the final assay volume being 60. mu.L. The buffer used to make up the final assay volume to 60. mu.L was 50mM MOPS, pH7.0, containing 1-5% DMSO. The reaction was terminated by the addition of 0.2-2 volumes of 0.75% phosphoric acid.

Peptide phosphorylation was accomplished as followsAfter peptides were collected on P8196 well filter plates (Whatman), scintillation counting was performed using a beta counter (TopCount). Total control cpm (C)⁺) Cpm (C) against background^-) Wells contained DMSO instead of compound. Background control (C)^-) The pores lack peptides. Assume total number (C)⁺) Subtract background (C)^-) The number is proportional to the initial reaction rate. The percent enzyme inhibition was calculated as 1- [ (cpm)_{Sample (I)}-C^-)/(C⁺-C^-)]X 100%. According to the four parameter logic equation Y ═ bottom + (top-bottom)/1 +10^ ((LogEC50-X)^＊Hill slope)), where X is the logarithm of the compound concentration and Y is the percent inhibition, using GraphPadPrism^TMDetermination of IC from enzyme inhibition-Compound concentration Curve₅₀The value is obtained.

In this assay, each of examples 1-92 in Table 1 exhibits an IC of greater than or equal to 3.0. mu.M₅₀The value is obtained.

Example 218

IGF-1 RTK enzyme assay

IGF-1 receptor tyrosine kinase assay utilizes the peptide substrate biotin-ahx-EQEDEPEGDYFEWLE-C (O) NH₂(Synpep) as a phosphoryl acceptor.

The assay was performed in 384 well black plates (Nunc). The kinase domain of the IGF-1 receptor was purchased from Upstate (cat. No. 14-465M). At 100. mu.M ATP and 20mM MgCl₂The enzyme was pre-activated on ice for 15min in the presence of (2). Compounds were diluted in DMSO prior to addition in the assay. In general, the assay is performed by incubating the enzyme (0.2-10nM) with or without inhibitor, 30. mu.M ATP, 5mM MgCl₂400nM peptide, incubated at 25 ℃ for 40min, the final assay volume being 20. mu.L. The assay buffer used was 50mM Tris-HCl, pH 7.5. The reaction was terminated by the addition of 10. mu.L of 0.15M EDTA.

After adding 25. mu.L of Eu-W1024-labeled anti-phosphotyrosine pTyr-100(PerkinElmer) antibody (final concentration 20nM) and 25. mu.L of streptavidin-APC (PerkinElmer, final concentration 20nM), the total volume was 80. mu.L, wherebyDetection of peptide phosphorylation was accomplished by homogeneous time-resolved fluorescence (HTRF). Both HTRF detection reagents were diluted in 50mM Tris-HCl, pH7.5 buffer, containing 0.5% BSA. Assay plates were incubated at 25 ℃ for 15min and Envision read in time-resolved fluorescence mode with the instrument set to excitation at 340nM and emission at 665 nM. Total control fluorescence Unit (C)⁺) Rfu (C) against background^-) Wells contained DMSO instead of compound. Background control (C)^-) The pores lack peptides. The percent enzyme inhibition was calculated as 1- [ (cpm)_{Sample (I)}-C^-)/(C⁺-C^-)]X 100%. According to the four parameter logic equation Y ═ bottom + (top-bottom)/1 +10^ ((LogEC50-X)_＊Hill slope)), where X is the logarithm of the compound concentration and Y is the percent inhibition, using GraphPad Prism^TMDetermination of IC from enzyme inhibition-Compound concentration Curve₅₀The value is obtained.

In this assay, each of examples 1-92 in Table 1 exhibits an IC of greater than or equal to 3.0. mu.M₅₀The value is obtained.

Example 219

CDK2 enzyme assay

The CDK2 kinase assay utilizes the peptide substrate biotin-ahx-ARRPMSPKKKA as the phosphoryl acceptor.

Assays were performed in 96-well U-bottom plates. CDK2 enzyme was purchased from PanVera. In general, the assay is carried out by incubating the enzyme (0.2-10nM) with or without inhibitor, 0.1-1. mu. Ci. gamma. at 25 ℃ with³³P-ATP(Perkin Elmer)、0.1-100μM ATP、0.1-10mMMgCl₂1-100 muM sodium orthovanadate, 1-10mM DTT and 1-100 muM peptide for 5-120min, and the final determination volume is 60 muL. The buffer used to make up the final assay volume to 60. mu.L was 50mM Tris-HCl, pH7.5, containing 1-5% DMSO and 0.1% BSA. The reaction was terminated by the addition of 0.2-2 volumes of 0.75% phosphoric acid. Compounds were diluted in DMSO prior to addition in the assay.

Detection of peptide phosphorylation was accomplished using a beta meter after peptide collection on a P8196 well filter plate (Whatman)Scintillation counting was performed with a counter (TopCount). Total control cpm (C)⁺) Cpm (C) against background^-) Wells contained DMSO instead of compound. Background control (C)^-) The pores lack peptides. Assume total number (C)⁺) Subtract background (C)^-) The number is proportional to the initial reaction rate. The percent enzyme inhibition was calculated as 1- [ (cpm)_{Sample (I)}-C^-)/(C⁺-C^-)]X 100%. According to the four parameter logic equation Y ═ bottom + (top-bottom)/1 +10^ ((LogEC50-X)^＊Hill slope)), where X is the logarithm of the compound concentration and Y is the percent inhibition, using GraphPadPrism^TMDetermination of IC from enzyme inhibition-Compound concentration Curve₅₀The value is obtained.

In this assay, each of examples 1-92 in Table 1 exhibits an IC of greater than or equal to 3.0. mu.M₅₀The value is obtained.

Example 220

VEGFR-2 TK enzyme assay

VEGFR-2 tyrosine kinase assay utilizing the peptide substrate biotin-ahx-EQEDEPEGDYFEWLE-C (O) NH₂As phosphoryl acceptors.

The kinase domain of VEGFR-2 was purchased from ProQuinase. At 100. mu.M ATP and 20mM MgCl₂The enzyme was pre-activated on ice for 15min in the presence of (2). Assays were performed in 96-well U-bottom plates. In general, the assay is performed by incubating the enzyme (0.2-10nM) with or without inhibitor, 30. mu.M ATP, 5mM MgCl at 25 ℃₂400nM peptide for 30min, final assay volume 20. mu.L. The buffer used to make up the final assay volume to 20. mu.L was 50mM Tris-HCl, pH 7.5. The reaction was terminated by the addition of 10. mu.L of 0.15M EDTA.

After addition of 25. mu.L of Eu-W1024-labeled anti-phosphotyrosine pTyr-100(PerkinElmer) antibody (final concentration 20nM) and 25. mu.L of streptavidin-APC (PerkinElmer, final concentration 20nM), in a total volume of 80. mu.L, detection of peptide phosphorylation was accomplished by homogeneous time-resolved fluorescence (HTRF). Both HTRF detection reagents were diluted in 50mM Tris-HCl, pH7.5 buffer containingThere was 0.5% BSA. Assay plates were incubated at 25 ℃ for 15min and Envision read in time-resolved fluorescence mode with the instrument set to excitation at 340nM and emission at 665 nM. Positive control (C)⁺) With negative control (C)^-) Wells contained DMSO instead of compound. Negative control (C)^-) The pores lack peptides. The percentage of enzyme inhibition was calculated as 1- [ (RFU)_{Sample (I)}-C^-)/(C⁺-C^-)]X 100%. According to the four parameter logic equation Y ═ bottom + (top-bottom)/1 +10^ ((LogEC50-X)^＊Hill slope)), where X is the logarithm of the compound concentration and Y is the percent inhibition, using GraphPad Prism^TMDetermination of IC from enzyme inhibition-Compound concentration Curve₅₀The value is obtained.

In this assay, each of examples 1-92 in Table 1 exhibits an IC of greater than or equal to 3.0. mu.M₅₀The value is obtained.

Example 221

In vitro cell proliferation

The compounds were tested for their ability to inhibit cell proliferation and viability. Using alamarBlue^TMReduction of metabolism (Biosource cat. No. dall 1100) was used to measure cell viability.

The antiproliferative activity of the compounds was studied using a panel of tumor cells: HCT-116 (human colorectal cancer cell line), BxPC-3 (human pancreatic cancer cell line), A549 (human lung cancer cell line), BT-549 (human breast cancer cell line), LNCaP (human prostate cancer cell line), and MIAPaca-2 (human pancreatic cancer cell line). These adherent cells (1,000-20,000) were plated in complete medium (RPMI-1640, DMEM, F12K or McCoy's 5A) in tissue culture-treated 96-well plates (Costar) containing 10% fetal bovine serum (Gibco) in a humidified incubator 37 ℃ with 95% O₂，5％CO₂The neutralization time is 18-24 h. The medium was removed and replaced with 90. mu.L of fresh medium. Compounds were diluted in medium containing 3% DMSO and added to cells. Incubation of alamarBlue with untreated cells plated 18h ago^TMReagent for 6h, background (C) was determined^-) Relative fluorescence units. Allowing untreated cellsOr cells containing the compound were incubated for 96 h. During the last 6h of the incubation period, alamarBlue was added to each well^TMReagent (10. mu.L) at 95% O in a humidified incubator at 37 ℃₂，5％CO₂And (4) carrying out incubation.

Measurement of alamarBlue in a fluorescence plate reader^TMThe instrument was set to excite at 530nm and emit at 590 nm. The percentage inhibition of cell growth was calculated as 1- [ (RFU)_{Sample (I)}-C^-)/(RFU_Untreated-C^-)]X 100%. According to the four parameter logic equation Y ═ bottom + (top-bottom)/1 +10^ ((LogEC50-X)^＊Hill slope)), using GraphPad Prism^TMDetermination of Compound IC from inhibition-Compound concentration Curve₅₀The value is obtained.

Each of the compounds in Table 1 exhibits an IC less than or equal to 3.0 μ M for one or more of the group of tumor cells₅₀The value is obtained. Specifically, examples 27, 35, 36, 48, 50, 54, 57, 58 and 60 had an IC of less than or equal to 3 μ M for at least one HCT-116, MIA Paca-2 or LNCaP cell using the assay conditions described above₅₀The value is obtained.

Pharmacological study of drug combinations

Although the compounds of the present invention may be used as single components, they may also be used as part of a combination therapy. For example, example 88 demonstrated anti-tumor activity in established human tumor xenografts in athymic mice when administered as a single component, including tumors derived from pancreatic and breast tissue (see the single component dose curve for example 88 in example 222-224, below). To evaluate the therapeutic efficacy and identify potential synergy of the combination of example 88 with other therapeutic agents, example with gemcitabine (Gemzar) was used^TM) Erlotinib (Tarceva)^TM) Or trastuzumab (Herceptin)^TM) Various animal studies were performed.

Gemcitabine, erlotinib and trastuzumab are therapeutic agents capable of treating a range of solid tumors. Gemcitabine, erlotinib andtrastuzumab has a mechanism of action that is distinct from each other and the compounds of the invention. Gemcitabine is a DNA synthesis inhibitor that inhibits ribonucleotide reductase. Erlotinib is an EGF receptor tyrosine kinase inhibitor that blocks the function of EGF growth factor. Gemcitabine and erlotinib are currently used to treat advanced pancreatic cancer. Trastuzumab is a recombinant humanized monoclonal antibody that binds to and blocks p185^HER2Receptor function. Trastuzumab is a first line treatment of metastatic breast cancer, a tumor that overexpresses HER2 protein.

Example 222

Example 88 alone and in combination with erlotinib was evaluated for anti-tumor activity against established human MiaPaCa-2 pancreatic xenodermal transplantation (preclinical pancreatic cancer model) in athymic mice. The compounds were administered according to the following protocol:

1) example 88 is i.p., b.i.d. daily for 10 days (days 1-10).

2) Erlotinib was 50mg/kg, p.o. daily for 14 days (days 1-14).

Antitumor activity was assessed by means of inhibition of tumor growth and evaluation of individual tumor regressions characterized by either partial (more than 50% reduction in tumor size) or complete (100% reduction in tumor size) responses.

Drug treatment started with an average tumor size of up to about 120mg (day 8). Mice bearing sc tumors were randomly grouped and the treatment group consisted of 8 mice. The median tumor size for each group at various time points in the study is listed in table a below.

TABLE A MiaPaCa-2 xenoskin transplantation model

	Carrier	Separate embodiment 88	Erlotinib alone	Example 88 with erlotinib
					Days of study	Median tumor size (mg)	Median tumor size (mg)	Median tumor size (mg)	Median tumor size (mg)
8	113	120	120	120
					10	171	162	144	162
13	192	170	192	162
					16	241	170	267	108
20	435	221	363	82
					23	609	209	507	69
27	988	368	700	88
					30	1282	486	908	101
34	1521	817	1224	148
					37	1770	1055	1296	225
42	2058	1629	1368	398

The MiaPaCa-2 tumor growth curves are shown in FIG. 1, wherein

A-represents the vector of example 88 and erlotinib;

o-represents erlotinib at a dose of 50mg/kg, p.o. for up to 14 days per day;

□ -representative of example 88 at a dose of 10mg/kg, i.p., b.i.d. for up to 10 days per day;

● -represents example 88 and erlotinib.

Previous dose response studies in this model found that example 88(10mg/kg) was a moderately effective dose for tumor growth inhibition in this model. Example 88(10mg/kg) produced 67% inhibition of tumor growth relative to vehicle-treated tumors on day 23. Erlotinib (50mg/kg) produced 16% inhibition of tumor growth on day 23. In contrast, the example 88/erlotinib combination produced 89% inhibition of tumor growth on day 23. Analysis of individual tumor regression behavior at day 42 showed that the example 88/erlotinib combination therapy produced 1 partial response and 2 complete responses in a total of 8 mice (table AA); however, none of the individual components produced a response.

TABLE AA.MIAPaCa-2 example 88 summary of response to xenogeneic transplantation

Type of response	Carrier	Example 9110mg/kg	Erlotinib 50mg/kg	Example 91+ erlotinib
					PR	0/8	0/8	0/8	1/8
CR	0/8	0/8	0/8	2/8

In summary, example 88(10mg/kg) when combined with erlotinib (50mg/kg) produced a higher anti-tumor response rate in the MiaPaCa-2 model than either example 88 or erlotinib administered as a single component.

Example 223

Example 88 alone and in combination with gemcitabine was evaluated for antitumor activity against a given human Mia PaCa-2 pancreatic xenodermal transplant (preclinical pancreatic cancer model) in athymic mice. The compounds were administered according to the following protocol:

1) example 88 is i.p., b.i.d. daily for 10 days (days 1-10).

2) Gemcitabine was 120mg/kg, i.p., q3d × 4 (days 1, 4, 7, 10).

Antitumor activity was assessed by means of inhibition of tumor growth and evaluation of individual tumor regressions characterized by either partial (more than 50% reduction in tumor size) or complete (100% reduction in tumor size) responses.

Drug treatment started with an average tumor size of up to about 120mg (day 8). Mice bearing sc tumors were randomly grouped and the treatment group consisted of 8 mice. The median tumor size for each group at various time points in the study is listed in table B below.

MiaPaCa-2 xenoskin transplantation model

	Carrier	Separate embodiment 88	Gemcitabine alone	Example 88 and Gemcitabine
					Days of study	Median tumor size (mg)	Median tumor size (mg)	Median tumor size (mg)	Median tumor size (mg)
8	113	120	120	126
					10	144	162	144	144
13	192	170	144	170
					16	192	170	153	98
20	295	221	221	69
					23	466	209	246	63
27	650	368	336	86
					30	757	486	472	149
34	972	817	675	251
					37	1132	1055	824	359
42	1353	1629	988	606

The MiaPaCa-2 tumor growth curve is shown in FIG. 2, wherein

■ -vector representing example 88 and gemcitabine;

a-represents example 88 at a dose of 10mg/kg, i.p., b.i.d. for up to 10 days per day;

□ -represents gemcitabine, at a dose of 120mg/kg, i.p., q3d × 4; and

o-represents example 88 and gemcitabine.

Previous dose response studies in this model found that example 88(10mg/kg) was a moderately effective dose for tumor growth inhibition in this model. Example 88(10mg/kg) produced 67% inhibition of tumor growth relative to vehicle-treated tumors on day 23. Gemcitabine (120mg/kg) produced a 60% inhibition of tumor growth on day 23. In contrast, the example 88/gemcitabine combination produced a 90% inhibition of tumor growth on day 23. Analysis of individual tumor regression behavior at day 42 indicated that the example 88/gemcitabine combination therapy produced 1 partial response and 1 complete response in a total of 8 mice; none of the individual components produced a response (Table BB).

TABLE BB. example 88 response summary of MIAPaCa-2 xenodermal transplantation

Type of response	Carrier	Example 8810mg/kg	Gemcitabine 120mg/kg	Example 88+ Gemcitabine
					PR	0/8	0/8	0/8	1/8
CR	0/8	0/8	0/8	1/8

In summary, example 88(10mg/kg), when combined with gemcitabine (120mg/kg), produced a higher anti-tumor response rate in the MiaPaCa-2 model than either example 88 or gemcitabine, administered as the single component.

Example 224

Trastuzumab is a first line treatment of metastatic breast cancer, a tumor that overexpresses HER2 protein. Example 88 in combination with trastuzumab was tested in the BT-474 mammary gland xenogeneic transplant model. Drug treatment started with an average tumor size of approximately 110mg (day 35 post-implantation). Mice bearing sc tumors were randomly grouped and the treatment group consisted of 10 mice. The compounds were administered according to the following protocol:

1) example 88 was 30mg/kg, i.p., b.i.d. for 3 days per day, then interrupted for 2 days for a total of 5 cycles;

2) trastuzumab at 10mg/kg, i.p. twice weekly for 4 weeks.

The median tumor size for each group at various time points in the study is listed in table C below.

BT-474 xenodermal transplantation model

	Carrier	Separate embodiment 88	Trastuzumab alone	Example 88 and trastuzumab
					Days of study	Median tumor size (mg)	Median tumor size (mg)	Median tumor size (mg)	Median tumor size (mg)
1	113	113	113	113
					4	170	120	133	152
7	190	152	132	189
					10	259	153	124	225
14	342	179	106	266
					17	394	149	80	284
21	504	142	54	384
					24	617	138	36	416
28		104	18	416

The BT-474 tumor growth curve in athymic SCID mice is shown in FIG. 3, where treatment was initiated day 1, where

■ -representative of the vector of example 88;

o-represents trastuzumab at a dose of 10mg/kg, i.p. twice weekly for 4 weeks;

a-represents example 88 at a dose of 30mg/kg, i.p., b.i.d. for 3 days per day, followed by 2 days of discontinuation for a total of 5 cycles;

□ -representative of example 88 and trastuzumab.

Example 88(30mg/kg) produced 77% inhibition of tumor growth relative to vehicle-treated tumors on day 24. Trastuzumab (10mg/kg) produced 33% inhibition of tumor growth on day 24. In contrast, the example 88/trastuzumab combination produced 94% inhibition of tumor growth on day 24. Analysis of individual tumor regression behavior at day 24 showed that example 88/trastuzumab combination therapy yielded 4 partial responses and 6 complete responses in a total of 10 mice (table CC); however, none of the individual components produced a response.

Table cc. bt-474 example 88 response summary of xenogeneic transplantation

Type of response	Carrier	Example 8810mg/kg	Trastuzumab 10mg/kg	Example 88+ Trastuzumab
					PR	0/10	0/10	0/10	4/10
CR	0/10	0/10	0/10	6/10

In summary, example 88(30mg/kg) when combined with trastuzumab (10mg/kg) produced superior complete tumor regression rates in the BT-474 xenodermal transplant model than either example 88 or trastuzumab administered as the single component.

The following procedure was used to prepare pharmaceutical formulations containing the compounds of the present invention.

Example 225

A pharmaceutical formulation containing 2.0mg/mL of example 88 (equivalent to 2.7mg/mL of the trihydrochloride salt of example 88) was prepared as follows.

The trihydrochloride salt of example 88 (1.350g) was dissolved in sterile water for injection (SWFI) with stirring. The SWFI may be degassed with sterile nitrogen prior to use. The amount of SWFI that dissolves example 88 is the amount that will dissolve the compound. In one embodiment, the amount of SWFI that dissolves the compound is more than 50% of the final volume, which may be 75% of the final volume.

D-mannitol was added to the solution and dissolved with stirring. The pH of the mixture is adjusted between 3.0 and 3.6 ± 0.1 with a small amount of alkaline solution, e.g. 1N NaOH, added gradually, either before the addition of mannitol or after the dissolution of mannitol. SWFI was then added to a final desired volume of 500 mL.

The solution was filtered through a 0.22 μm PVDF filter into a container. The solution can be prefiltered using 0.45 μm VDF. Finally, 10.25mL of the filtered solution was transferred to a 20mL vial (borosilicate glass type I vial) which had been flushed with sterile nitrogen prior to use. The filled vials were then sealed with Fluorotec B2-40 stoppers. Vials can be stored at 8 ℃ or below 8 ℃ and above freezing.

Example 226

Using a procedure similar to example 225, a 7mg/mL + -0.3 solution of example 88 can be prepared, wherein the final solution has a pH of between pH2.5 and 3.0 + -0.1 and a final volume of 35mL in each vial. Vials can be stored at 8 ℃ or below 8 ℃ and above freezing.

Example 227

Diluents for use in combination with the example 225 formulations or other formulations containing the compounds of the invention may be prepared as follows.

SWFI (490mL) degassed with sterile nitrogen was transferred to a vessel and the pH adjusted to pH11.0 to 11.4. + -. 0.1. SWFI was then added to a final desired volume of 500 mL. The solution was then filtered through a 0.22 μm PVDF filter to a 0.22 μm PVDF filter into a vessel. Finally, 10.25mL of the filtered solution was transferred to a 20mL vial (borosilicate glass type I vial) which had been flushed with sterile nitrogen prior to use.

Example 228

Using a procedure similar to example 227, a diluent having a ph of 11.0 to 11.4 ± 0.1 was prepared, wherein 65mL of the filtered solution was transferred to a 100mL vial.

Example 229

Prior to administration, the formulation of example 225 was diluted with the diluent of example 227, where the contents of the diluent (10.25mL) were transferred in small amounts to a vial containing example 225, such that the final concentration of example 88 was 1mg/mL and the final pH was 5.5 ± 0.1. The solutions were combined, showed no precipitation, and were stable between 15 and 30 ℃ for a time sufficient for dosage preparation and administration. Such a period of time is at least 1 to 6 hours.

Example 230

Prior to administration, the formulation of example 226 was diluted with the diluent of example 228, wherein the contents of the diluent (65mL) were transferred in small amounts to a vial containing example 226, such that the final concentration of example 88 was 2mg/mL and the final pH was 4.5 ± 0.1. The solutions were combined, showed no precipitation, and were stable between 15 and 30 ℃ for a time sufficient for dosage preparation and administration. Such a period of time is at least 1 to 6 hours and may be as long as 24 hours.

While the invention has been described and illustrated with reference to certain embodiments thereof, those skilled in the art will appreciate that various changes, modifications and substitutions can be made without departing from the spirit and scope of the invention. For example, effective dosages other than those described herein may be useful as a consequence of the differences in responsiveness of subjects being treated for Aurora kinase-mediated conditions. Likewise, the specific pharmacological responses observed may vary according to and depending upon the particular active compound selected or whether there is a pharmaceutical carrier present, as well as the type of formulation and mode of administration employed, and such expected differences or variations in results are contemplated in accordance with the objects and practices of the present invention.

Claims (16)

1. A compound of formula (Ib):

wherein

G¹Is a 1H-indazol-6-yl group,

wherein G is¹Is a compound which is unsubstituted and which is,

G²selected from the group consisting of: phenyl and pyridin-2-yl groups,

wherein G is²Is unsubstituted or substituted with at least one substituent selected from the group consisting of: a methyl group, a methoxy group and a trifluoromethyl group,

q is a piperazin-1-yl group,

or a pharmaceutically acceptable salt thereof.

2.6-piperazin-1-yl-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide, or a pharmaceutically acceptable salt thereof.

3. The compound of claim 2, wherein the compound is 6-piperazin-1-yl-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide, or a hydrochloride salt thereof.

4. The compound of claim 2, wherein the compound is 6-piperazin-1-yl-2- (2-trifluoromethylphenylamino) -1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide.

2- (3-methylpyridin-2-ylamino) -6-piperazin-1-yl-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide, or a pharmaceutically acceptable salt thereof.

6. The compound of claim 5, wherein the compound is 2- (3-methylpyridin-2-ylamino) -6-piperazin-1-yl-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide, or a hydrochloride salt thereof.

7. The compound of claim 5, wherein the compound is 2- (3-methylpyridin-2-ylamino) -6-piperazin-1-yl-1H-benzimidazole-5-carboxylic acid (1H-indazol-6-yl) -amide.

8. A pharmaceutical composition comprising a compound of any one of claims 1-7 and a pharmaceutically acceptable carrier.

9. A pharmaceutical composition comprising a compound of any one of claims 1-7 and an additional therapeutic agent selected from the group consisting of an antimetabolite, a protein tyrosine kinase inhibitor, and an antibody.

10. The pharmaceutical composition according to claim 9, wherein the additional therapeutic agent is selected from the group consisting of gemcitabine, erlotinib, and trastuzumab.

11. Use of a compound according to any one of claims 1 to 7 in the manufacture of a medicament for the treatment of cancer.

12. Use of a compound of any one of claims 1 to 7 in the manufacture of a medicament for inhibiting Aurora kinase activity, wherein said use for inhibiting Aurora kinase activity comprises contacting a cell of a subject in which inhibition of Aurora kinase a or B is desired with a compound of any one of claims 1 to 7.

13. Use of a compound of any one of claims 1-7 in the manufacture of a medicament for treating an Aurora kinase-mediated disorder in a subject in need thereof.

14. The use according to claim 13 wherein the Aurora kinase-mediated disorder is cancer.

15. The use according to claim 14, wherein the cancer is selected from the group consisting of: colorectal cancer, ovarian cancer, breast cancer, gastric cancer, prostate cancer, brain cancer, bone cancer, bladder cancer, head and neck cancer, lung cancer, kidney cancer, pancreatic cancer, sarcoma, leukemia, and lymphoma.

16. The use according to claim 15, wherein the cancer is selected from the group consisting of: breast cancer, colorectal cancer, and pancreatic cancer.