Note: Descriptions are shown in the official language in which they were submitted.
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IMIDAZOPYRAZINES AS PROTEIN KINASE INHIBITORS
Field of the Invention
The present invention relates to imidazo[1,2-a]pyrazine compounds useful as
protein kinase inhibitors, regulators or modulators, pharmaceutical
compositions
containing the compounds, and methods of treatment using the compounds and
compositions to treat diseases such as, for example, cancer, inflammation,
arthritis,
viral diseases, neurodegenerative diseases such as Alzheimer's disease,
cardiovascular diseases, and fungal diseases. The present compounds are
especially
10. useful as Aurora kinase inhibitors.
Background of the Invention
Protein kinases are a family of enzymes that catalyze phosphorylation of
proteins, in particular the hydroxyl group of specific tyrosine, serine, or
threonine
residues in proteins. Protein kinases are pivotal in the regulation of a wide
variety of
cellular processes, including metabolism, cell proliferation, cell
differentiation, and cell
survival. Uncontrolled proliferation is a hallmark of cancer cells, and can be
manifested by a deregulation of the cell division cycle in one of two ways -
making
stimulatory genes hyperactive or inhibitory genes inactive. Protein kinase
inhibitors,
regulators or modulators alter the function of kinases such as cyclin-
dependent
kinases (CDKs), mitogen activated protein kinase (MAPK/ERK), glycogen synthase
kinase 3 (GSK3beta), Checkpoint (Chk) (e.g., CHK-1, CHK-2 etc.) kinases, AKT
kinases, JNK, and the like. Examples of protein kinase inhibitors are
described in
W002/22610 Al and by Y. Mettey et al in J. Med. Chem., (2003) 46 222-236.
The cyclin-dependent kinases are serine/threonine protein kinases, which are
the driving force behind the cell cycle and cell proliferation. Misregulation
of CDK
function occurs with high frequency in many important solid tumors. Individual
CDK's,
such as, CDK1, CDK2, CDK3, CDK4, CDK5, CDK6 and CDK7, CDK8 and the like,
perform distinct roles in cell cycle progression and can be classified as
either G1, S, or
G2M phase enzymes. CDK2 and CDK4 are of particular interest because their
activities are frequently misregulated in a wide variety of human cancers.
CDK2
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activity is required for progression through G1 to the S phase of the cell
cycle, and
CDK2 is one of the key components of the G1 checkpoint. Checkpoints serve to
maintain the proper sequence of cell cycle events and allow the cell to
respond to
insults or to proliferative signals, while the loss of proper checkpoint
control in cancer
cells contributes to tumorgenesis. The CDK2 pathway influences tumorgenesis at
the
level of tumor suppressor function (e.g. p52, RB, and p27) and oncogene
activation
(cyclin E). Many reports have demonstrated that both the coactivator, cyclin
E, and
the inhibitor, p27, of CDK2 are either over- or underexpressed, respectively,
in breast,
colon, nonsmall cell lung, gastric, prostate, bladder, non-Hodgkin's lymphoma,
ovarian, and other cancers. Their altered expression has been shown to
correlate
with increased CDK2 activity levels and poor overall survival. This
observation makes
CDK2 and its regulatory pathways compelling targets for the development of
cancer
treatments.
A number of adenosine 5'-tnphosphate (ATP) competitive small organic
molecules as well as peptides have been reported in the literature as CDK
inhibitors
for the potential treatment of cancers. U.S. 6,413,974, col. 1, line 23- col.
15, line 10
offers a good description of the various CDKs and their relationship to
various types of
cancer. Flavopiridol (shown below) is a nonselective CDK inhibitor that is
currently
undergoing human clinical trials, A. M. Senderowicz et al, J. Clin. Oncol.
(1998) 16,
2986-2999.
1H3
HO~
HO
CI
OH O
Other known inhibitors of CDKs include, for example, olomoucine (J. Vesely et
al, Eur. J. Biochem., (1994) 224, 771-786) and roscovitine (I. Meijer et al,
Eur. J.
Biochem., (1997) 243, 527-536). U.S. 6,107,305 describes certain pyrazolo[3,4-
b]
pyridine compounds as CDK inhibitors. An illustrative compound from the '305
patent
is:
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0
N
/ N NJ
H
K. S. Kim et al, J. Med. Chem. 45 (2002) 3905-3927 and WO 02/10162
disclose certain aminothiazole compounds as CDK inhibitors.
Imidazopyrazines are known. For example, U.S. 6,919,341 (the disclosure of
which is
incorporated herein by reference) and US2005/0009832 disclose various
imidazopyrazines. Also being mentioned are the following: W02005/047290;
US2005/095616; W02005/039393; W02005/019220; W02004/072081;
W02005/014599; W02005/009354; W02005/005429; W02005/085252;
US2005/009832; US2004/220189; W02004/074289; W02004/026877;
W02004/026310; W02004/022562; W02003/089434; W02003/084959;
W02003/051346; US2003/022898; W02002/060492; W02002/060386;
W02002/028860; JP (1986)61-057587; J. Burke et al., J. Biological Chem., Vol.
278(3), 1450-1456 (2003); and F. Bondavalli et a!, J. Med. Chem., Vol. 45 22 ,
4875-
4887 (2002).
Also made reference to are US 2004/0220189 (published November 4, 2004);
US 2005/0009832 (published January 13, 2005); US 2006/0084650 (published April
20, 2006) which describe kinase inhibitors, and
US 2006/0106023 (published May 18, 2006) which describe imidazopyrazines as
cyclin dependent kinase inhibitors. In addition, US 2007/0117804 (published
May 24,
2007), describes imidazopyrazines as protein kinase inhibitors of the
following
structure:
A compound of the Formula:
R1 R2
N\\/ N
R3.N,H
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or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof,
wherein:
R is H, CN, -NR5R6, cycloalkyl, cycloalkenyl, heterocyclenyl, heteroaryl,
-C(0)NR5R6, -N(R5)C(O)R6, heterocyclyl, heteroaryl substituted with (CH2)13
NR5R6, unsubstituted alkyl, or alkyl substituted with one or more moieties
which
can be the same or different each moiety being independently selected from
the group consisting of -OR5, heterocyclyl,
-N(R5)C(O)N(R5R6), -N(R5)-C(0)OR6, -(CH2)1_3-N(R5R6) and -NR5R6;
R1 is H, halo, aryl or heteroaryl, wherein each of said aryl and heteroaryl
can be
unsubstituted or substituted with one or more moieties which can be the same
or different each moiety being independently selected from the group
consisting
of halo, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heterocyclyl,
-CH2OR5, -C(O)NR5R6, -C(O)OH, -C(O)NH2,
-NR5R6 (wherein the R5 and R6, together with the N of said
-NR5R6, form a heterocyclyl ring), -S(O)R5, -S(02)R5, -CN, -CHO, -SRS, -
C(O)OR5, -C(O)R5 and -OR5;
R2 is H, halo, aryl, arylalkyl or heteroaryl, wherein each of said aryl,
arylalkyl and
heteroaryl can be unsubstituted or optionally independently be substituted
with
one or more moieties which can be the same or different each moiety being
independently selected from the group consisting of halo, amide, alkyl,
alkenyl,
alkynyl, cycloalkyl, aryl,
-C(O)OH, -C(O)NH2, -NR5R6 (wherein the R5 and R6, together with the N of
said -NR5R6, form a heterocyclyl ring), -CN, arylalkyl,
-CH2OR5, -S(0)R5, -S(02)R5, -CN, -CHO, -SR5, -C(O)ORS, -C(O)R5, heteroaryl
and heterocyclyl;
R3 is H, alkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein:
- said alkyl shown above for R3 can be unsubstituted or substituted with one
or
more moieties which can be the same or different each moiety being
independently selected from the group consisting of -ORS, alkoxy,
heteroaryl, and -NR5R6;
- said aryl shown above for R3 is unsubstituted, or optionally substituted, or
optionally fused, with halo, heteroaryl, heterocyclyl, cycloalkyl or
heteroarylalkyl, wherein each of said heteroaryl, heterocyclyl, cycloalkyl
and heteroarylalkyl can be unsubstituted or optionally independently
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substituted with one or more moieties which can be the same or different
each moiety being independently selected from alkyl, -ORS, -N(R5R6)
and
-S(02)R5; and
5 - said heteroaryl shown above for R3 can be unsubstituted or optionally
substituted, or optionally fused, with one or more moieties which can be
the same or different with each moiety being independently selected
from the group consisting of halo, amino, alkoxycarbonyl, -OR5, alkyl, -
CHO, - NR5R6, -S(02)N(RSR6),
-C(O)N(R5R6), -SR5, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl,
heterocyclenyl, and heterocyclyl;
R5 is H, alkyl, aminoalkyl, aryl, heteroaryl, heterocyclyl or cycloalkyl; and
R6 is H, alkyl, aryl, arylalkyl, heteroaryl, heterocyclyl or cycloalkyl;
further wherein in any -NR5R6 in Formula I, said R5 and R6 can optionally be
joined
together with the N of said -NR5R6 to form a cyclic ring.
Another series of protein kinases are those that play an important role as a.
checkpoint in cell cycle progression. Checkpoints prevent cell cycle
progression at
inappropriate times, such as in response to DNA damage, and maintain the
metabolic
balance of cells while the cell is arrested, and in some instances can induce
apoptosis
(programmed cell death) when the requirements of the checkpoint have not been
met.
Checkpoint control can occur in the G1 phase (prior to DNA synthesis) and in
G2,
prior to entry into mitosis.
One series of checkpoints monitors the integrity of the genome and, upon
sensing DNA damage, these "DNA damage checkpoints" block cell cycle
progression
in G, & G2 phases, and slow progression through S phase. This action enables
DNA
repair processes to complete their tasks before replication of the genome and
subsequent separation of this genetic material into new daughter cells takes
place.
Inactivation of CHK1 has been shown to transduce signals from the DNA-damage
sensory complex to inhibit activation of the cyclin B/Cdc2 kinase, which
promotes
mitotic entry, and abrogate G<sub>2</sub> arrest induced by DNA damage inflicted by
anticancer agents or endogenous DNA damage, as well as result in preferential
killing
of the resulting checkpoint defective cells. See, e.g., Peng et al., Science,
277, 1501-
1505 (1997); Sanchez et al., Science, 277, 1497-1501 (1997), Nurse, Cell, 91,
865-
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867 (1997); Weinert, Science, 277, 1450-1451 (1997); Walworth et at., Nature,
363,
368-371 (1993); and Al-Khodairy et al., Molec. Biol. Cell., 5, 147-160 (1994).
Selective manipulation of checkpoint control in cancer cells could afford
broad
utilization in cancer chemotherapeutic and radiotherapy regimens and may, in
addition, offer a common hallmark of human cancer "genomic instability" to be
exploited as the selective basis for the destruction of cancer cells. A number
of
factors place CHK1 as a pivotal target in DNA-damage checkpoint control. The
elucidation of inhibitors of this and functionally related kinases such as
CDS1/CHK2, a
kinase recently discovered to cooperate with CHK1 in regulating S phase
progression
(see Zeng et al., Nature, 395, 507-510 (1998); Matsuoka, Science, 282, 1893-
1897
(1998)), could provide valuable new therapeutic entities for the treatment of
cancer.
Another group of kinases are the tyrosine kinases. Tyrosine kinases can be of
the receptor type (having extracellular, transmembrane and intracellular
domains) or
the non-receptor type (being wholly intracellular). Receptor-type tyrosine
kinases are
comprised of a large number of transmembrane receptors with diverse biological
activity. In fact, about 20 different subfamilies of receptor-type tyrosine
kinases have
been identified. One tyrosine kinase subfamily, designated the HER subfamily,
is
comprised of EGFR (HER1), HER2, HERS and HER4. Ligands of this subfamily of
receptors identified so far include epithelial growth factor, TGF-alpha,
amphiregulin,
HB-EGF, betacellulin and heregulin. Another subfamily of these receptor-type
tyrosine kinases is the insulin subfamily, which includes INS-R, IGF-IR, IR,
and IR-R.
The PDGF subfamily includes the PDGF-alpha and beta receptors, CSFIR, c-kit
and
FLK-I1. The FLK family is comprised of the kinase insert domain receptor
(KDR), fetal
liver kinase-1 (FLK-1), fetal liver kinase-4 (FLK-4) and the fms-like tyrosine
kinase-1
(fit-1). For detailed discussion of the receptor-type tyrosine kinases, see
Plowman et
al., DN&P 7(6): 334-339, 1994.
At least one of the non-receptor protein tyrosine kinases, namely, LCK, is
believed to mediate the transduction in T-cells of a signal from the
interaction of a cell-
surface protein (Cd4) with a cross-linked anti-Cd4 antibody. A more detailed
discussion of non-receptor tyrosine kinases is provided in Bolen, Oncogene, 8,
2025-
2031(1993). The non-receptor type of tyrosine kinases is also comprised of
numerous subfamilies, including Src, Frk, Btk, Csk, Abl, Zap70, Fes/Fps, Fak,
Jak,
Ack, and LIMK. Each of these subfamilies is further sub-divided into varying
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receptors. For example, the Src subfamily is one of the largest and includes
Src, Yes,
Fyn, Lyn, Lck, Blk, Hck, Fgr, and Yrk. The Src subfamily of enzymes has been
linked
to oncogenesis. For a more detailed discussion of the non-receptor type of
tyrosine
kinases, see Bolen, Oncogene, 8:2025-2031 (1993).
In addition to its role in cell-cycle control, protein kinases also play a
crucial role
in angiogenesis, which is the mechanism by which new capillaries are formed
from
existing vessels. When required, the vascular system has the potential to
generate
new capillary networks in order to maintain the proper functioning of tissues
and
organs. In the adult, however, angiogenesis is fairly limited, occurring only
in the
process of wound healing and neovascularization of the endometrium during
menstruation. On the other hand, unwanted angiogenesis is a hallmark of
several
diseases, such as retinopathies, psoriasis, rheumatoid arthritis, age-related
macular
degeneration, and cancer (solid tumors). Protein kinases which have been shown
to
be involved in the angiogenic process include three members of the growth
factor
receptor tyrosine kinase family; VEGF-R2 (vascular endothelial growth factor
receptor
2, also known as KDR (kinase insert domain receptor) and as FLK 1); FGF-R
(fibroblast growth factor receptor); and TEK (also known as Tie-2).
VEGF-R2, which is expressed only on endothelial cells, binds the potent
angiogenic growth factor VEGF and mediates the subsequent signal transduction
through activation of its intracellular kinase activity. Thus, it is expected
that direct
inhibition of the kinase activity of VEGF-R2 will result in the reduction of
angiogenesis
even in the presence of exogenous VEGF (see Strawn et al, Cancer Research, 56,
3540-3545 (1996)), as has been shown with mutants of VEGF-R2 which fail to
mediate signal transduction. Millauer et al, Cancer Research, 56, 1615-1620
(1996).
Furthermore, VEGF-R2 appears to have no function in the adult beyond that of
mediating the angiogenic activity of VEGF. Therefore, a selective inhibitor of
the
kinase activity of VEGF-R2 would be expected to exhibit little toxicity.
Similarly, FGFR binds the angiogenic growth factors aFGF and bFGF and
mediates subsequent intracellular signal transduction. Recently, it has been
suggested that growth factors such as bFGF may play a critical role in
inducing
angiogenesis in solid tumors that have reached a certain size. Yoshiji et al.,
Cancer
Research, 57, 3924-3928 (1997). Unlike VEGF-R2, however, FGF-R is expressed in
a number of different cell types throughout the body and may or may not play
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important roles in other normal physiological processes in the adult.
Nonetheless,
systemic administration of a small molecule inhibitor of the kinase activity
of FGF-R
has been reported to block bFGF-induced angiogenesis in mice without apparent
toxicity. Mohammad et al., EMBO Journal, 17, 5996-5904 (1998).
TEK (also known as Tie-2) is another receptor tyrosine kinase expressed only
on endothelial cells which has been shown to play a role in angiogenesis. The
binding
of the factor angiopoietin-1 results in autophosphorylation of the kinase
domain of
TEK and results in a signal transduction process which appears to mediate the
interaction of endothelial cells with peri-endothelial support cells, thereby
facilitating
the maturation of newly formed blood vessels. The factor angiopoietin-2, on
the other
hand, appears to antagonize the action of angiopoietin-1 on TEK and disrupts
angiogenesis. Maisonpierre et al.,. Science, 277, 55-60 (1997).
The kinase, JNK, belongs to the mitogen-activated protein kinase (MAPK)
superfamily. JNK plays a crucial role in inflammatory responses, stress
responses,
cell proliferation, apoptosis, and tumongenesis. JNK kinase activity can be
activated
by various stimuli, including the proinflammatory cytokines (TNF-alpha and
interleukin-
1), lymphocyte costimulatory receptors (CD28 and CD40), DNA-damaging
chemicals,
radiation, and Fas signaling. Results from the JNK knockout mice indicate that
JNK is
involved in apoptosis induction and T helper cell differentiation.
Pim-1 is a small serine/threonine kinase. Elevated expression levels of Pim-1
have been detected in lymphoid and myeloid malignancies, and recently Pim-1
was
identified as a prognostic marker in prostate cancer. K. Peltola, "Signaling
in Cancer:
Pim-1 Kinase and its Partners", Annales Universitatis Turkuensis, Sarja - Ser.
D Osa
- Tom. 616, (August 30, 2005),
htt://ki 'asto.utu.fi/iulkaisu alvelut/annaalit/2004/D616.html. Pim-1 acts as
a cell
survival factor and may prevent apoptosis in malignant cells. K. Petersen Shay
et al.,
Molecular Cancer Research 3:170-181 (2005).
Yet another group of kinases are Aurora kinases. Aurora kinases (Aurora-A,
Aurora-B, Aurora-C) are serine/threonine protein kinases that have been
implicated in
human cancer, such as colon, breast and other solid tumors. Aurora-A (also
sometimes referred to as AIK) is believed to be involved in protein
phosphorylation
events that regulate the cell cycle. Specifically, Aurora-A may play a role in
controlling
the accurate segregation of chromosomes during mitosis. Misregulation of the
cell
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cycle can lead to cellular proliferation and other abnormalities. In human
colon cancer
tissue, Aurora-A, Aurora-B, Aurora-C have been found to be over-expressed
(see,
Bischoff et al., EMBO J., 17:3052-3065 (1998); Schumacher et al., J. Cell
Biol.
143:1635-1646 (1998); Kimura et al., J. Biol. Chem., 272:13766-13771 (1997)).
There is a need for effective inhibitors of protein kinases, especially Aurora
kinases, in order to treat or prevent disease states associated with abnormal
cell
proliferation. Moreover, it is desirable to have kinase inhibitors, especially
small-
molecule compounds that may be readily synthesized.
Summary of the Invention
In its many embodiments, the present invention provides a novel class of
imidazo[1,2-a]pyrazine compounds, methods of preparing such compounds,
pharmaceutical compositions comprising one or more such compounds, methods of
preparing pharmaceutical formulations comprising one or more such compounds,
and
methods of treatment, prevention, inhibition or amelioration of one or more
diseases
associated with protein kinases using such compounds or pharmaceutical
compositions.
In one aspect, the present invention provides compounds represented by
Formula Z:
R^
A~
RN~
NY N
NHR3
Formula Z
or a pharmaceutically acceptable salt, solvate, ester, prodrug or stereoisomer
thereof,
wherein:
R is H, halo or alkyl;
R3 is heteroaryl-X, wherein X is (heterocyclyl)alkyl- wherein said
heterocyclyl
can be unsubstituted or optionally substituted with 1-4 alkyl moieties;
A is -aryl- , -heteroaryl-, -N(R')-aryl- or -N(R')-heteroaryl- , wherein each
of
said aryl and heteroaryl can be independently unsubstituted or optionally
substituted
with one or more substituents, which substituents can be the same or
different, each
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substituent being independently selected from the group consisting of alkyl, -
NO2,
halo, hydroxy, trihaloalkyl, alkoxy, and dialkylamino;
. NRIR2 -,)yOH
RA is -(CH2)1 -heteroaryl, 0 or 0 , wherein said
heteroaryl can optionally be fused with an aryl, wherein each of said aryl and
5 heteroaryl can independently be optionally substituted with one or more
moieties each
moiety being independently selected from the group consisting of trihaloalkyl,
-NO2,
halo, hydroxyalkyl, alkoxyalkyl and dialkylamino;
R1 is H or alkyl;
R2 is H, hydroxyalkyl-, arylalkyl-, heteroaryl, aryl, heteroarylalkyl-, alkyl,
10 dialkylaminoalkyl-, alkylaminoalkyl-, cycloalkylalkyl-, cycloalkyl,
heterocyclylalkyl- or heterocyclyl, wherein said aryl and aryl of arylalkyl
can be
unsubstituted or substituted with one or more moieties independently selected
from the group consisting of trihaloalkyl,.-N02, halo, hydroxyalkyl,
alkoxyalkyl,
dialkylamino and heterocyclylalkyl-, wherein said heterocyclylalkyl can be
unsubstituted or'substituted with alkyl or-S02NH2; said heteroaryl and
heteroaryl of heteroarylalkyl can be unsubstituted or substituted with one or
more moieties, each moiety being independently selected from the group
consisting of hydroxyalkyl, alkoxy, alkyl, halo, hydroxyl, and -N02; and said
cycloalkyl is unsubstituted or substituted with hydroxyl; or
R1 and R2 together with the N to which each is attached, form a
-i-N N-Y
heterocyclic group selected from the group consisting of
--N S -k-N 0 -~-N Y"
~-- ~--, and , wherein
Y is alkoxyalkyl, hydroxyalkyl, dialkylaminoalkyl or alkyl, further wherein
Y is hydroxyl.
In another aspect, the present invention provides compounds represented by
Formula I:
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RA
A~
RN
N.zN
NHR3
Formula I
or a pharmaceutically acceptable salt, solvate, ester, prodrug or stereoisomer
thereof,
wherein:
R is halo or alkyl;
R3 is heteroaryl-X, wherein X is heterocyclylalkyl- wherein said heterocyclyl
can
be unsubstituted or optionally substituted with 1-4 alkyl moieties;
A is heteroaryl, wherein said heteroaryl can be unsubstituted or optionally
substituted with one or more substituents, which substituents can be the same
or
different, each substituent being independently selected from the group
consisting of
alkyl, -NO2, halo, hydroxy, trihaloalkyl, alkoxy, and dialkylamino;
NR RZ , ~OH
RA is II -(CH2)1A-heteroaryl, 0 or 0 , wherein said
heteroaryl can optionally be fused with an aryl, wherein each of said aryl and
heteroaryl can independently be optionally substituted with one or more
moieties each
moiety being independently selected from the group consisting of trihaloalkyl,
-N02,
halo, hydroxyalkyl, alkoxyalkyl and dialkylamino;
R1 is H or alkyl;
R2 is H, hydroxyalkyl-, arylalkyl-, heteroaryl, aryl, heteroarylalkyl-, alkyl,
dialkylaminoalkyl-, alkylaminoalkyl-, cycloalkylalkyl-, cycloalkyl,
heterocyclylalkyl- or heterocyclyl, wherein said aryl and aryl of arylalkyl
can be
unsubstituted or substituted with one or more moieties independently selected
from the group consisting of trihaloalkyl, -NO2, halo, hydroxyalkyl,
alkoxyalkyl,
dialkylamino and heterocyclylalkyl-, wherein said heterocyclylalkyl can be
unsubstituted or substituted with alkyl or -SO2NH2; said heteroaryl and
heteroaryl of heteroarylalkyl can be unsubstituted or substituted with one or
more moieties, each moiety being independently selected from the group
consisting of hydroxyalkyl, alkoxy, alkyl, halo, hydroxyl, and -N02; and said
cycloalkyl is unsubstituted or substituted with hydroxyl; or
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Wand R2 together with the N to which each is attached, form a
+N -Y
heterocyclic group selected from the group consisting of `_...1 ,
--N S -1-N 0 -~-N Y
\-j , \---/ and , wherein
Y is alkoxyalkyl, hydroxyalkyl, dialkylaminoalkyl or alkyl, further wherein
Y is hydroxyl.
The compounds of Formulas I and Z can be useful as protein kinase inhibitors.
The compounds of Formulas I and Z can also be useful as Aurora kinase
inhibitors.
The compounds of Formulas I and Z can be useful in the treatment and
prevention of
proliferative diseases, for example, cancer, inflammation and arthritis,
neurodegenerative diseases such Alzheimer's disease, cardiovascular diseases,
viral
diseases and fungal diseases.
Detailed Description
In an embodiment, the present invention provides imidazopyrazine compounds,
especially imidazo[1,2-a]pyrazine compounds, which are represented by
structural
Formula Z, or pharmaceutically acceptable salts, solvates, esters or prodrug
thereof,
wherein the various moieties are as described above.
In another embodiment, the present invention provides imidazopyrazine
compounds, especially imidazo[1,2-a]pyrazine compounds, which are represented
by
structural Formula I, or pharmaceutically acceptable salts, solvates, esters
or prodrug
thereof, wherein the various moieties are as described above.
The following embodiments are intended for, independently, Formula I where
applicable, as well as for Formula Z where applicable. The embodiments are
independent of one another:
In another embodiment, A is aryl, as described above.
In another embodiment, A is heteroaryl, as described above.
In another embodiment, A is -NR'-heteroaryl, as described above.
In another embodiment, A is heteroaryl, as described above.
In another embodiment, A is aryl, as described above.
In another embodiment, RA is -(CH2)1-4-heteroaryl.
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// II NRIR2
In another embodiment, RA is o
OH
In another embodiment, RA is 0
In another embodiment, R is H.
In another embodiment, R is alkyl.
In another embodiment, R is methyl.
In another embodiment, R is halo.
In another embodiment, R1 is H.
In another embodiment, R1 is alkyl.
In another embodiment, R is methyl.
In another embodiment, R is halo.
In another embodiment, R3 is heteroaryl-(unsubstituted heterocyclyl).
In another embodiment, R3 is heteroaryl-(heterocyclyl(methyl)).
In another embodiment, R3 is heteroaryl-(heterocyclyl(methyl)2).
In another embodiment, R3 is thiazolyl substituted with heterocyclyl which is
substituted with 1-3 alkyl.
In another embodiment, R3 is thiazolyl substituted with heterocyclyl which is
un
substituted.
In another embodiment, R3 is thiazolyl substituted with piperidyl which may be
optionally substituted with 1-3 alkyl.
Non-limiting examples of compounds of Formulas I and Z include the various
compounds shown in the Tables below, or a pharmaceutically acceptable salt,
solvate, ester or prodrug thereof.
As used above, and throughout this disclosure, the following terms, unless
otherwise indicated, shall be understood to have the following meanings,
including
any possible substitutions of the stated groups or moieties:
"Patient" includes both human and animals.
"Mammal" means humans and other mammalian animals.
"Alkyl" means an aliphatic hydrocarbon group which may be straight or
branched and comprising about 1 to about 20 carbon atoms in the chain.
Preferred
alkyl groups contain about 1 to about 12 carbon atoms in the chain. More
preferred
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14
alkyl groups contain about 1 to about 6 carbon atoms in the chain. Branched
means
that one or more, lower alkyl groups such as methyl, ethyl or propyl, are
attached to a
linear alkyl chain. "Lower alkyl" means a group having about 1 to about 6
carbon
atoms in the chain which may be straight or branched. "Alkyl" may be
unsubstituted or
optionally substituted by one or more substituents which may be the same or
different,
each substituent being independently selected from the group consisting of
halo, alkyl,
aryl, cycloalkyl, cyano, hydroxy, alkoxy, alkylthio, amino, oxime (e.g., =N-
OH), -
NH(alkyl), -NH(cycloalkyl), -N(alkyl)2, -O-C(O)-alkyl, -O-C(O)-aryl,
-O-C(O)-cycloalkyl, carboxy and -C(O)O-alkyl. Non-limiting examples of
suitable alkyl
groups include methyl, ethyl, n-propyl, isopropyl and t-butyl.
"Alkenyl" means an aliphatic hydrocarbon group containing.at least one carbon-
carbon double bond and which may be straight or branched and comprising about
2 to
about 15 carbon atoms in the chain. Preferred alkenyl groups have about 2 to
about
12 carbon atoms in the chain; and more preferably about 2 to about 6 carbon
atoms in
the chain. Branched means that one or more, lower alkyl groups such as methyl,
ethyl
or propyl, are attached to a linear alkenyl chain, "Lower alkenyl" means about
2 to
about 6 carbon atoms in the chain which may be straight or branched. "Alkenyl"
may
be unsubstituted or optionally substituted by one or more substituents which
may be
the same or different, each substituent being independently selected from the
group
consisting of halo, alkyl. aryl, cycloalkyl, cyano, alkoxy and -S(alkyl). Non-
limiting
examples of suitable alkenyl groups include ethenyl, propenyl, n-butenyl, 3-
methylbut-
2-enyl, n-pentenyl, octenyl and decenyl.
"Alkylene" means a difunctional group obtained by removal of a hydrogen atom
from an alkyl group that is defined above. Non-limiting examples of alkylene
include
methylene, ethylene and propylene.
"Alkynyl" means an aliphatic hydrocarbon group containing at least one carbon-
carbon triple bond and which may be straight or branched and comprising about
2 to
about 15 carbon atoms in the chain. Preferred alkynyl groups have about 2 to
about
12 carbon atoms in the chain; and more preferably about 2 to about 4 carbon
atoms in
the chain. Branched means that one or more, lower alkyl groups such as methyl,
ethyl
or propyl, are attached to a linear alkynyl chain. "Lower alkynyl" means about
2 to
about 6 carbon atoms in the chain which may be straight or branched. Non-
limiting
examples of suitable alkynyl groups include ethynyl, propynyl, 2-butynyl and 3-
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methylbutynyl. "Alkynyl" may be unsubstituted or optionally substituted by one
or more
substituents which may be the same or different, each substituent being
independently selected from the group consisting of alkyl, aryl and
cycloalkyl.
"Aryl" means an aromatic monocyclic or multicyclic ring system comprising
5 about 6 to about 14 carbon atoms, preferably about 6 to about 10 carbon
atoms. The
aryl group can be optionally substituted with one or more "ring system
substituents"
which may be the same or different, and are as defined herein. Non-limiting
examples
of suitable aryl groups include phenyl and naphthyl.
"Bridged cyclic ring" is a hydrocarbon ring such as cycloalkyl, cyclenyl, or
aryl
10 or heteroatom containing ring such as, heterocyclyl, heterocyclenyl, or
heteroaryl as
described herein, that contains a bridge, which is a valence bond or an atom
or an
unbranched chain of atoms connecting two different parts of the ring. The two
tertiary
carbon atoms connected through the bridge are termed "bridgeheads".
"Heteroaryl" means an aromatic monocyclic or multicyclic ring system
15 comprising about 5 to about 14 ring atoms, preferably about 5 to about 10
ring atoms,
in which one or more of the ring atoms is an element other than carbon, for
example
nitrogen, oxygen or sulfur, alone or in combination. Preferred heteroaryls
contain
about 5 to about 6 ring atoms. The "heteroaryl" can be optionally substituted
by one or
more "ring system substituents" which may be the same or different, and are as
defined herein. The prefix aza, oxa or thia before the heteroaryl root name
means that
at least a nitrogen, oxygen or sulfur atom respectively, is present as a ring
atom. A
nitrogen atom of a heteroaryl can be optionally oxidized to the corresponding
N-oxide.
"Heteroaryl" may also include a heteroaryl as defined above fused to an aryl
as
defined above. Non-limiting examples of suitable heteroaryls include pyridyl;
pyrazinyl,
furanyl, thienyl, pyrimidinyl, pyridone (including N-substituted pyridones),
isoxazolyl,
isothiazolyl, oxazolyl, thiazolyl, pyrazolyl, furazanyl, pyrrolyl, pyrazolyl,
triazolyl, 1,2,4-
thiadiazolyl, pyrazinyl, pyridazinyl, quinoxalinyl, phthalazinyl, oxindolyl,
imidazo[1,2-
a]pyridinyl, imidazo[2,1-b]thiazolyl, benzofurazanyl, indolyl, azaindolyl,
benzimidazolyl,
benzothienyl, quinolinyl, imidazolyl, thienopyridyl, quinazolinyl,
thienopyrimidyl,
pyrrolopyridyl, imidazopyridyl, isoquinolinyl, benzoazaindolyl, 1,2,4-traziyl,
benzothiazolyl and the like. The term "heteroaryl" also refers to partially
saturated
heteroaryl moieties such as, for example, tetrahydroisoquinolyl,
tetrahydroquinolyl and
the like.
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"Aralkyl" or "arylalkyl" means an aryl-alkyl- group in which the aryl and
alkyl are
as previously described. Preferred aralkyls comprise a lower alkyl group. Non-
limiting
examples of suitable aralkyl groups include benzyl, 2-phenethyl and
naphthalenylmethyl. The bond to the parent moiety is through the alkyl.
"Alkylaryl" means an alkyl-aryl- group in which the alkyl and aryl are as
previously described. Preferred alkylaryls comprise a lower alkyl group. Non-
limiting
example of a suitable alkylaryl group is tolyl. The bond to the parent moiety
is through
the aryl.
"Cycloalkyl" means a non-aromatic mono- or multicyclic ring system comprising
about 3 to about 10 carbon atoms, preferably about 5 to about 10 carbon atoms.
Preferred cycloalkyl rings contain about 5 to about 7 ring atoms. The
cycloalkyl can be
optionally substituted with one or more "ring system substituents" which may
be the
same or different, and are as defined above. Non-limiting examples of suitable
monocyclic cycloalkyls include cyclopropyl, cyclopentyl, cyclohexyl,
cycloheptyl and
the like. Non-limiting examples of suitable multicyclic cycloalkyls include 1-
decalinyl,
norbomyl, adamantyl and the like.
"Cycloalkylalkyl" means a cycloalkyl moiety as defined above linked via an
alkyl
moiety (defined above) to a parent core. Non-limiting examples of suitable
cycloalkylalkyls include cyclohexylmethyl, adamantylmethyl and the like.
"Cycloalkenyl" means a non-aromatic mono or multicyclic ring system
comprising about 3 to about 10 carbon atoms, preferably about 5 to about 10
carbon
atoms which contain at least one carbon-carbon double bond. Preferred
cycloalkenyl
rings contain about 5 to about 7 ring atoms. The cycloalkenyl can be
optionally
substituted with one or more "ring system substituents" which may be the same
or
different, and are as defined above. Non-limiting examples of suitable
monocyclic
cycloalkenyls include cyclopentenyl, cyclohexenyl, cyclohepta-1,3-dienyl, and
the like.
Non-limiting example of a suitable multicyclic cycloalkenyl is norbornylenyl.
"Cycloalkenylalkyl" means a cycloalkenyl moiety as defined above linked via an
alkyl moiety (defined above) to a parent core. Non-limiting examples of
suitable
cycloalkenylalkyls include cyclopentenylmethyl, cyclohexenylmethyl and the
like.
"Halogen" means fluorine, chlorine, bromine, or iodine. Preferred are
fluorine,
chlorine and bromine.
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"Ring system substituent" means a substituent attached to an aromatic or non-
aromatic ring system which, for example, replaces an available hydrogen on the
ring
system. Ring system substituents may be the same or different, each being
independently selected from the group consisting of alkyl, alkenyl, alkynyl,
aryl,
heteroaryl, aralkyl, alkylaryl, heteroaralkyl, heteroarylalkenyl,
heteroarylalkynyl,
alkylheteroaryl, hydroxy, hydroxyalkyl, alkoxy, aryloxy, aralkoxy, acyl,
aroyl, halo,
nitro, cyano, carboxy, alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl,
alkylsulfonyl,
arylsulfonyl, heteroarylsulfonyl, alkylthio, arylthio, heteroarylthio,
aralkylthio,
heteroaralkylthio, cycloalkyl, heterocyclyl, amide, -CHO, -O-C(0)-alkyl, -O-
C(0)-aryl, -
O-C(O)-cycloalkyl, -C(=N-CN)-NH2, -C(=NH)-NH2, -C(=NH)-NH(alkyl), oxime (e.g.,
=N-OH), Y1Y2N-, Y1Y2N-alkyl-, Y1Y2NC(O)-, Y1Y2NSO2- and -SO2NYIY2, wherein Y(
and Y2 can be the same or different and are independently selected from the
group
consisting of hydrogen, alkyl, aryl, cycloalkyl, and aralkyl. "Ring system
substituent"
may also mean a single moiety which simultaneously replaces two available
hydrogen
on two adjacent carbon atoms (one H on each carbon) on a ring system. Examples
of
such moiety are methylene dioxy, ethylenedioxy, -C(CH3)2- and the like which
form
moieties such as, for example:
Co
~I
o -0 and
"Heteroarylalkyl" means a heteroaryl moiety as defined above linked via an
alkyl moiety (defined above) to a parent core. Non-limiting examples of
suitable
heteroaryls include 2-pyridinylmethyl, quinolinylmethyl and the like.
"Heterocyclyl" means a non-aromatic saturated monocyclic or multicyclic ring
system comprising about 3 to about 10 ring atoms, preferably about 5 to about
10 ring
atoms, in which one or more of the atoms in the ring system is an element
other than
carbon, for example nitrogen, oxygen or sulfur, alone or in combination. There
are no
adjacent oxygen and/or sulfur atoms present in the ring system. Preferred
heterocyclyls contain about 5 to about 6 ring atoms. The prefix aza, oxa or
thia before
the heterocyclyl root name means that at least a nitrogen, oxygen or sulfur
atom
respectively is present as a ring atom. Any -NH in a heterocyclyl ring may
exist
protected such as, for example, as an -N(Boc), -N(CBz), -N(Tos) group and the
like;
such protections are also considered part of this invention. The heterocyclyl
can be
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18
optionally substituted by one or more "ring system substituents" which may be
the
same or different, and are as defined herein. The nitrogen or sulfur atom of
the
heterocyclyl can be optionally oxidized to the corresponding N-oxide, S-oxide
or S,S-
dioxide. Non-limiting examples of suitable monocyclic heterocyclyl rings
include
piperidyl, pyrrolidinyl, piperazinyl, morpholinyl, thiomorpholinyl,
thiazolidinyl, 1,4-
dioxanyl, tetrahydrofuranyl, tetrahydrothiophenyl, lactam, lactone, and the
like.
"Heterocyclyl" may also mean a single moiety (e.g., carbonyl) which
simultaneously
replaces two available hydrogen on the same carbon atom on a ring system.
Example
of such moiety is pyrrolidone:
H
N
0
"Heterocyclylalkyl" means a heterocyclyl moiety as defined above linked via an
alkyl moiety (defined above) to a parent core. Non-limiting examples of
suitable
.heterocyclylalkyls include piperidinylmethyl, piperazinylmethyl and the like.
"Heterocyclenyl" means a non-aromatic monocyclic or multicyclic ring system
comprising about 3 to about 10 ring atoms, preferably about 5 to about 10 ring
atoms,
in which one or more of the atoms in the ring system is an element other than
carbon,
for example nitrogen, oxygen or sulfur atom, alone or in combination, and
which
contains at least one carbon-carbon double bond or carbon-nitrogen double
bond.
There are no adjacent oxygen and/or sulfur atoms present in the ring system.
Preferred heterocyclenyl rings contain about 5 to about 6 ring atoms. The
prefix aza,
oxa or thia before the heterocyclenyl root name means that at least a
nitrogen, oxygen
or sulfur atom respectively is present as a ring atom. The heterocyclenyl can
be
optionally substituted by one or more ring system substituents, wherein "ring
system
substituent" is as defined above. The nitrogen or sulfur atom of the
heterocyclenyl can
be optionally oxidized to the corresponding N-oxide, S-oxide or S,S-dioxide.
Non-
limiting examples of suitable heterocyclenyl groups include 1,2,3,4
tetrahydropyridinyl, 1,2-dihydropyridinyl, 1,4-dihydropyridinyl, 1,2,3,6-
tetrahydropyridinyl, 1,4,5,6-tetrahydropyrimidinyl, 2-pyrrolinyl, 3-
pyrrolinyl, 2-
imidazolinyl, 2-pyrazolinyl, dihydroimidazolyl, dihydrooxazolyl,
dihydrooxadiazolyl,
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dihydrothiazolyl, 3,4-dihydro-2H-pyranyl, dihydrofuranyl,
fluorodihydrofuranyl, 7-
oxabicYcloI2.2.1ShePtenY1, dihydrothiophenyl, dihydrothiopyranyl, and the
like.
"Heterocyclenyl" may also mean a single moiety (e.g., carbonyl) which
simultaneously
replaces two available hydrogen on the same carbon atom on a ring system.
Example
of such moiety is pyrrolidinone:
H
N
0
"Heterocyclenylalkyl" means a heterocyclenyl moiety as defined above linked
via an alkyl moiety (defined above) to a parent core.
It should be noted that in hetero-atom containing ring systems of this
invention,
there are no hydroxyl groups on carbon atoms adjacent to a N, 0 or S, as well
as
there are no N or S groups on carbon adjacent to another heteroatom. Thus, for
example, in the ring:
4
Cl/
5
N
H
there is no -OH attached directly to carbons marked 2 and 5.
It should also be noted that tautomeric forms such as, for example, the
moieties:
N 0 Cal
H and N OH
are considered equivalent in certain embodiments of this invention.
"Alkynylalkyl" means an alkynyl-alkyl- group in which the alkynyl and alkyl
are
as previously described. Preferred alkynylalkyls contain a lower alkynyl and a
lower
alkyl group. The bond to the parent moiety is through the alkyl. Non-limiting
examples
of suitable alkynylalkyl groups include propargylmethyl.
"Heteroaralkyl" means a heteroaryl-alkyl- group in which the heteroaryl and
alkyl are as previously described. Preferred heteroaralkyls contain a lower
alkyl group.
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Non-limiting examples of suitable aralkyl groups include pyridylmethyl, and
quinolin-3-
ylmethyl. The bond to the parent moiety is through the alkyl.
"Spiro ring systems" have two or more rings' linked by one common atom.
Preferred Spiro ring systems include spiroheteroaryl, spiroheterocyclenyl,
5 spiroheterocyclyl, spirocycloalkyl, spirocyclenyl, and spiroaryl. The Spiro
ring systems
can be optionally substituted by one or more ring system substituents, wherein
"ring
system substituent" is as defined above. Non-limiting examples of suitable
spiro ring
9 10 1
8
C)09
systems include 7 6 4
a s z
HN8 5 I 'I g I3
spiro[4.5]decane, 8-azaspiro[4.5]dec-2-ene, and
10 spiro[4.4]nona-2,7-diene.
"Hydroxyalkyl" means a HO-alkyl- group in which alkyl is as previously
defined.
Preferred hydroxyalkyls contain lower alkyl. Non-limiting examples of suitable
hydroxyalkyl groups include hydroxymethyl and 2-hydroxyethyl.
"Acyl" means an H-C(O)-, alkyl-C(O)- or cycloalkyl-C(0)-, group in which the
15 various groups are as previously described. The bond to the parent moiety
is through
the carbonyl. Preferred acyls contain a lower alkyl. Non-limiting examples of
suitable
acyl groups include formyl, acetyl and propanoyl.
"Aroyl" means an aryl-C(O)- group in which the aryl group is as previously
described. The bond to the parent moiety is through the carbonyl. Non-limiting
20 examples of suitable groups include benzoyl and 1- naphthoyl.
"Alkoxy" means an alkyl-0- group in which the alkyl group is as previously
described. Non-limiting examples of suitable alkoxy groups include methoxy,
ethoxy,
n-propoxy, isopropoxy and n-butoxy. The bond to the parent moiety is through
the
ether oxygen.
"Aryloxy" means an aryl-0- group in which the aryl group is as previously
described. Non-limiting examples of suitable aryloxy groups include phenoxy
and
naphthoxy. The bond to the parent moiety is through the ether oxygen.
"Aralkyloxy" means an aralkyl-O- group in which the aralkyl group is as
previously described. Non-limiting examples of suitable aralkyloxy groups
include
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benzyloxy and 1- or 2-naphthalenemethoxy. The bond to the parent moiety is
through
the ether oxygen.
"Alkylthio" means an alkyl-S- group in which the alkyl group is as previously
described. Non-limiting examples of suitable alkylthio groups include
methylthio and
ethylthio. The bond to the parent moiety is through the sulfur.
"Arylthio" means an aryl-S- group in which the aryl group is as previously
described. Non-limiting examples of suitable arylthio groups include
phenylthio and
naphthylthio. The bond to the parent moiety is through the sulfur.
"Aralkylthio" means an aralkyl-S- group in which the aralkyl group is as
previously described. Non-limiting example of a suitable aralkylthio group is
benzylthio. The bond to the parent moiety is through the sulfur.
"Alkoxycarbonyl" means an alkyl-O-CO- group. Non-limiting examples of
suitable alkoxycarbonyl groups include methoxycarbonyl and ethoxycarbonyl. The
bond to the parent moiety is through the carbonyl.
"Aryloxycarbonyl" means an aryl-O-C(O)- group. Non-limiting examples of
suitable aryloxycarbonyl groups include phenoxycarbonyl and naphthoxycarbonyl.
The
bond to the parent moiety is through the carbonyl.
"Aralkoxycarbonyl" means an aralkyl-O-C(O)- group. Non-limiting example of a
suitable aralkoxycarbonyl group is benzyloxycarbonyl. The bond to the parent
moiety
is through the carbonyl.
"Alkylsulfonyl" means an alkyl-S(02)- group. Preferred groups are those in
which the alkyl group is lower alkyl. The bond to the parent moiety is through
the
sulfonyl.
"Arylsulfonyl" means an aryl-S(02)- group. The bond to the parent moiety is
through the sulfonyl.
The term "substituted" means that one or more hydrogen on the designated
atom is replaced with a selection from the indicated group, provided that the
designated atom's normal valency under the existing circumstances is not
exceeded,
and that the substitution results in a stable compound. Combinations of
substituents
and/or variables are permissible only if such combinations result in stable
compounds.
By "stable compound' or "stable structure" is meant a compound that is
sufficiently
robust to survive isolation to a useful degree of purity from a reaction
mixture, and
formulation into an efficacious therapeutic agent.
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22
The term "optionally substituted" means optional substitution with the
specified
groups, radicals or moieties.
The term "purified", "in purified form" or "in isolated and purified form" for
a
compound refers to the physical state of said compound after being isolated
from a
synthetic process (e.g. from a reaction mixture), or natural source or
combination
thereof. Thus, the term "purified", "in purified form" or "in isolated and
purified form" for
a compound refers to the physical state of said compound after being obtained
from a
purification process or processes described herein or well known to the
skilled artisan
(e.g., chromatography, recrystallization and the like) , in sufficient purity
to be
characterizable by standard analytical techniques described herein or well
known to
the skilled artisan.
It should also be noted that any carbon as well as heteroatom with unsatisfied
valences in the text, schemes, examples and Tables herein is assumed to have
the
sufficient number of hydrogen atom(s) to satisfy the valences.
When a functional group in a compound is termed "protected", this means that
the group is in modified form to preclude undesired side reactions at the
protected site
when the compound is subjected to a reaction. Suitable protecting groups will
be
recognized by those with ordinary skill in the art as well as by reference to
standard
textbooks such as, for example, T. W. Greene et al, Protective Groups in
organic
Synthesis (1991), Wiley, New York.
When any variable (e.g., aryl, heterocycle, R2, etc.) occurs more than one
time
in any constituent or in Formula I, its definition on each occurrence is
independent of
its definition at every other occurrence.
As used herein, the term "composition" is intended to encompass a product
comprising the specified ingredients in the specified amounts, as well as any
product
which results, directly or indirectly, from combination of the specified
ingredients in the
specified amounts.
Prodrugs and solvates of the compounds of the invention are also
contemplated herein. A discussion of prodrugs is provided in T. Higuchi and V.
Stella,
Pro-drugs as Novel Delivery Systems (1987) 14 of the A.C.S. Symposium Series,
and
in Bioreversible Carriers in Drug Design, (1987) Edward B. Roche, ed.,
American
Pharmaceutical Association and Pergamon Press. The term "prodrug" means a
compound (e.g., a drug precursor) that is transformed in vivo to yield a
compound of
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23
Formula (I) or a pharmaceutically acceptable salt, hydrate or solvate of the
compound.
The transformation may occur by various mechanisms (e.g., by metabolic or
chemical
processes), such as, for example, through hydrolysis in blood. A discussion of
the
use of prodrugs is provided by T. Higuchi and W. Stella, "Pro-drugs as Novel
Delivery
Systems," Vol. 14 of the A.C.S. Symposium Series, and in Bioreversible
Carriers in
Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and
Pergamon Press, 1987.
For example, if a compound of Formula (I) or a pharmaceutically acceptable
salt, hydrate or solvate of the compound contains a carboxylic acid functional
group, a
prodrug can comprise an ester formed by the replacement of the hydrogen atom
of
the acid group with a group such as, for example, (C1-C8)alkyl, (C2-
C12)alkanoyloxymethyl, 1-(alkanoyloxy)ethyl having from 4 to 9 carbon atoms, 1-
methyl-1-(alkanoyloxy)-ethyl having from 5 to 10 carbon atoms,
alkoxycarbonyloxymethyl having from 3 to 6 carbon atoms, 1-
(alkoxycarbonyloxy)ethyl
having from 4 to 7 carbon atoms, 1-methyl-1-(alkoxycarbonyloxy)ethyl having
from 5
to 8 carbon atoms, N-(alkoxycarbonyl)aminomethyl having from 3 to 9 carbon
atoms,
1-(N-(alkoxycarbonyl)amino)ethyl having from 4 to 10 carbon atoms, 3-
phthalidyl, 4-
crotonolactonyl, gamma-butyrolacton-4-yl, di-N,N-(C1-C2)alkylamino(C2-C3)alkyl
(such
asp-dimethylaminoethyl), carbamoyl-(C1-C2)alkyl, N,N-di (C,-C2)alkylcarbamoyl-
(C1-
C2)alkyl and piperidino-, pyrrolidino- or morpholino(C2-C3)alkyl, and the
like.
Similarly, if a compound of Formula (I) contains an alcohol functional group,
a
prodrug can be formed by the replacement of the hydrogen atom of the alcohol
group
with a group such as, for example, (C1-C6)alkanoyloxymethyl, 1-((C1-
C6)alkanoyloxy)ethyl, 1-methyl-1-((C1-C6)alkanoyloxy)ethyl, (C1-
C6)alkoxycarbonyloxymethyl, N-(C1-C6)alkoxycarbonylaminomethyl, succinoyl, (C1-
C6)alkanoyl, a-amino(C1-C4)alkanyl, arylacyl and a-aminoacyl, or a-aminoacyl-a-
aminoacyl, where each a-aminoacyl group is independently selected from the
naturally
occurring L-amino acids, P(O)(OH)2, -P(O)(O(C1-C6)alkyl)2 or glycosyl (the
radical
resulting from the removal of a hydroxyl group of the hemiacetal form of a
carbohydrate), and the like.
If a compound of Formula (I) incorporates an amine functional group, a prodrug
can be formed by the replacement of a hydrogen atom in the amine group with a
.
group such as, for example, R-carbonyl, RO-carbonyl, NRR'-carbonyl where R and
R'
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24
are each independently (C1-C1o)alkyl, (C3-C7) cycloalkyl, benzyl, or R-
carbonyl is a
natural a-aminoacyl or natural a-aminoacyl, -C(OH)C(O)0Y1 wherein Y1 is H, (C,-
C6)alkyl or benzyl, -C(OY2)Y3 wherein Y2 is (C1-C4) alkyl and Y3 is (Ci-
C6)alkyl,
carboxy (C1-C6)alkyl, amino(Ci-C4)alkyl or mono-N-or di-N,N-(C1-
C6)alkylaminoalkyl,
-C(Y4)Y5 wherein Y4 is H or methyl and Y5 is mono-N- or di-N,N-(C,-
C6)alkylamino
morpholino, piperidin-1-yl or pyrrolidin-1-yl, and the like.
One or more compounds of the invention may exist in unsolvated as well as
solvated forms with pharmaceutically acceptable solvents such as water,
ethanol, and
the like, and it is intended that the invention embrace both solvated and
unsolvated
forms. "Solvate" means a physical association of a compound of this invention
with
one or more solvent molecules. This physical association involves varying
degrees of
ionic and covalent bonding, including hydrogen bonding. In certain instances
the
solvate will be capable of isolation, for example when one or more solvent
molecules
are incorporated in the crystal lattice of the crystalline solid. "Solvate"
encompasses
both solution-phase and isolatable solvates. Non-limiting examples of suitable
solvates include ethanolates, methanolates, and the like. "Hydrate" is a
solvate
wherein the solvent molecule is H20.
One or more compounds of the invention may optionally be converted to a
solvate. Preparation of solvates is generally known. Thus, for example, M.
Caira et al,
J. Pharmaceutical Sci., 93(3), 601-611 (2004) describes the preparation of the
solvates of the antifungal fluconazole in ethyl acetate as well as from water.
Similar
preparations of solvates, hemisolvate, hydrates and the like are described by
E. C.
van Tonder et al, AAPS PharmSciTech., 50), article 12 (2004); and A. L.
Bingham et
al, Chem. Commun., 603-604 (2001). A typical, non-limiting, process involves
dissolving the inventive compound in desired amounts of the desired solvent
(organic
or water or mixtures thereof) at a higher than ambient temperature, and
cooling the
solution at a rate sufficient to form crystals which are then isolated by
standard
methods. Analytical techniques such as, for example I. R. spectroscopy, show
the
presence of the solvent (or water) in the crystals as a solvate (or hydrate).
"Effective amount" or "therapeutically effective amount" is meant to describe
an
amount of compound or a composition of the present invention effective in
inhibiting
the above-noted diseases and thus producing the desired therapeutic,
ameliorative,
inhibitory or preventative effect.
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The compounds of Formulas I and Z can form salts which are also within the
scope of this invention. Reference to a compound of Formulas I and Z herein is
understood to include reference to salts thereof, unless otherwise indicated.
The term
"salt(s)", as employed herein, denotes acidic salts formed with inorganic
and/or
5 organic acids, as well as basic salts formed with inorganic and/or organic
bases. In
addition, when a compound of Formulas I and Z contains both a basic moiety,
such
as, but not limited to a pyridine or imidazole, and an acidic moiety, such as,
but not
limited to a carboxylic acid, zwitterions ("inner salts") may be formed and
are included
within the term "salt(s)" as used herein. Pharmaceutically acceptable (i.e.,
non-toxic,
10 physiologically acceptable) salts are preferred, although other salts are
also useful.
Salts of the compounds of the Formulas I and Z may be formed, for example, by
reacting a compound of Formulas I and Z with an amount of acid or base, such
as an
equivalent amount, in a medium such as one in which the salt precipitates or
in an
aqueous medium followed by lyophilization.
15 Exemplary acid addition salts include acetates, ascorbates, benzoates,
benzenesulfonates, bisulfates, borates, butyrates, citrates, camphorates,
camphorsulfonates, fumarates, hydrochlorides, hydrobromides, hydroiodides,
lactates,
maleates, methanesulfonates, naphthalenesulfonates, nitrates, oxalates,
phosphates,
propionates, salicylates, succinates, sulfates, tartarates, thiocyanates,
20 toluenesulfonates (also known as tosylates,) and the like. Additionally,
acids which are
generally considered suitable for the formation of pharmaceutically useful
salts from
basic pharmaceutical compounds are discussed, for example, by P. Stahl et al,
Camille G. (eds.) Handbook of Pharmaceutical Salts. Properties, Selection and
Use.
(2002) Zurich: Wiley-VCH; S. Berge et al, Journal of Pharmaceutical Sciences
(1977)
25 660) 1-19; P. Gould, International J. of Pharmaceutics (1986) 33 201-217;
Anderson
et al, The Practice of Medicinal Chemistry (1996), Academic Press, New York;
and in
The Orange Book (Food & Drug Administration, Washington, D.C. on their
website).
These disclosures are incorporated herein by reference thereto.
Exemplary basic salts include ammonium salts, alkali metal salts such as
sodium, lithium, and potassium salts, alkaline earth metal salts such as
calcium and
magnesium salts, salts with organic bases (for example, organic amines) such
as,
dicyclohexylamines, t-butyl amines, and salts with amino acids such as
arginine,
lysine and the like. Basic nitrogen-containing groups may be quartemized with
agents
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26
such as lower alkyl halides (e.g. methyl, ethyl, and butyl chlorides, bromides
and
iodides), dialkyl sulfates (e.g. dimethyl, diethyl, and dibutyl sulfates),
long chain
halides (e.g. decyl, lauryl, and stearyl chlorides, bromides and iodides),
aralkyl halides
(e.g. benzyl and phenethyl bromides), and others.
All such acid salts and base salts are intended to be pharmaceutically
acceptable salts within the scope of the invention and all acid and base salts
are
considered equivalent to the free forms of the corresponding compounds for
purposes
of the invention.
Pharmaceutically acceptable esters of the present compounds include the
following groups: (1) carboxylic acid esters obtained by esterification of the
hydroxy
groups, in which the non-carbonyl moiety of the carboxylic acid portion of the
ester
grouping is selected from straight or branched chain alkyl (for example,
acetyl, n-
propyl, t-butyl, or n-butyl), alkoxyalkyl (for example, methoxymethyl),
aralkyl (for
example, benzyl), aryloxyalkyl (for example, phenoxymethyl), aryl (for
example,
phenyl optionally substituted with, for example, halogen, C1.4alkyl, or
C1.4alkoxy or
amino); (2) sulfonate esters, such as alkyl- or aralkylsulfonyl (for example,
methanesulfonyl); (3) amino acid esters (for example, L-valyl or L-isoleucyl);
(4)
phosphonate esters and (5) mono-, di- or triphosphate esters. The phosphate
esters
may be further esterified by, for example, a C1_20 alcohol or reactive
derivative thereof,
or by a 2,3-di (Cr,24)acyl glycerol.
Compounds of Formula I, and salts, solvates, esters and prodrugs thereof, may
exist in their tautomeric form (for example, as an amide or imino ether). All
such
tautomeric forms are contemplated herein as part of the present invention.
The compounds of Formula (I) may contain asymmetric or chiral centers, and,
therefore, exist in different stereoisomeric forms. It is intended that all
stereoisomeric
forms of the compounds of Formula (I) as well as mixtures thereof, including
racemic
mixtures, form part of the present invention. In addition, the present
invention
embraces all geometric and positional isomers. For example, if a compound of
Formula (I) incorporates a double bond or a fused ring, both the cis- and
trans-forms,
as well as mixtures, are embraced within the scope of the invention.
Diastereomeric mixtures can be separated into their individual diastereomers
on the basis of their physical chemical differences by methods well known to
those
skilled in the art, such as, for example, by chromatography and/or fractional
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27
crystallization. Enantiomers can be separated by converting the enantiomeric
mixture
into a diastereomeric mixture by reaction with an appropriate optically active
compound (e.g., chiral auxiliary such as a chiral alcohol or Mosher's acid
chloride),
separating the diastereomers and converting (e.g., hydrolyzing) the individual
diastereomers to the corresponding pure enantiomers. Also, some of the
compounds
of Formula (I) may be atropisomers (e.g., substituted biaryls) and are
considered as
part of this invention. Enantiomers can also be separated by use of chiral
HPLC
column.
It is also possible that the compounds of Formula (I) may exist in different
tautomeric forms, and all such forms are embraced within the scope of the
invention.
Also, for example, all keto-enol and imine-enamine forms of the compounds are
included in the invention.
All stereoisomers (for example, geometric isomers, optical isomers and the
like)
of the present compounds (including those of the salts, solvates, esters and
prodrugs
of the compounds as well as the salts, solvates and esters of the prodrugs),
such as
those which may exist due to asymmetric carbons on various substituents,
including
enantiomeric forms (which may exist even in the absence of asymmetric
carbons),
rotameric forms, atropisomers, and diastereomeric forms, are contemplated
within the
scope of this invention, as are positional isomers (such as, for example, 4-
pyridyl and
3-pyridyl). (For example, if a compound of Formula (I) incorporates a double
bond or a
fused ring, both the cis- and trans-forms, as well as mixtures, are embraced
within the
scope of the invention. Also, for example, all keto-enol and imine-enamine
forms of
the compounds are included in the invention.) Individual stereoisomers of the
compounds of the invention may, for example, be substantially free of other
isomers,
or may be admixed, for example, as racemates or with all other, or other
selected,
stereoisomers. The chiral centers of the present invention can have the S or R
configuration as defined by the IUPAC 1974 Recommendations. The use of the
terms
"salt", "solvate", "ester", "prodrug" and the like, is intended to equally
apply to the salt,
solvate, ester and prodrug of enantiomers, stereoisomers, rotamers, tautomers,
positional isomers, racemates or prodnigs of the inventive compounds.
The present invention also embraces isotopically-labelled compounds of the
present invention which are identical to those recited herein, but for the
fact that one
or more atoms are replaced by an atom having an atomic mass or mass number
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28
different from the atomic mass or mass number usually found in nature.
Examples of
isotopes that can be incorporated into compounds of the invention include
isotopes of
hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, such as
2H,
3H, 13C, 14C, 15N, 180, 170, 31p, 32P, 35S, 18F, and 36CI, respectively.
Certain isotopically-labelled compounds of Formula (I) (e.g., those labeled
with
3H and 14C) are useful in compound and/or substrate tissue distribution
assays.
Tritiated (i.e., 3H) and carbon-14 (i.e., 14C) isotopes are particularly
preferred for their
ease of preparation and delectability. Further, substitution with heavier
isotopes such
as deuterium (i.e., 2H) may afford certain therapeutic advantages resulting
from
greater metabolic stability (e.g., increased in vivo half-life or reduced
dosage
requirements) and hence may be preferred in some circumstances. Isotopically
labeled compounds of Formula (I) can generally be prepared by following
procedures
analogous to those disclosed in the Schemes and/or in the Examples herein
below, by
substituting an appropriate isotopically labeled reagent for a non-
isotopically labeled
reagent.
Polymorphic forms of the compounds of Formula I, and of the salts, solvates,
esters and prodrugs of the compounds of Formula I, are intended to be included
in the
present invention.
The compounds according to the invention have pharmacological properties; in
particular, the compounds of Formulas I and Z can be inhibitors, regulators or
modulators of protein kinases. Non-limiting examples of protein kinases that
can be
inhibited, regulated or modulated include cyclin-dependent kinases (CDKs),
such as,
CDK1, CDK2, CDK3, CDK4, CDK5, CDK6 and CDK7, CDK8, mitogen activated
protein kinase (MAPK/ERK), glycogen synthase kinase 3 (GSK3beta), Pim-1
kinases,
Chk kinases (such as Chk1 and Chk2), tyrosine kinases, such as the HER
subfamily
(including, for example, EGFR (HER1), HER2, HER3 and HER4), the insulin
subfamily (including, for example, INS-R, IGF-IR, IR, and IR-R), the PDGF
subfamily
(including, for example, PDGF-alpha and beta receptors, CSFIR, c-kit and FLK-
II), the
FLK family (including, for example, kinase insert domain receptor (KDR), fetal
liver
kinase-1(FLK-1), fetal liver kinase-4 (FLK-4) and the fms-like tyrosine kinase-
1 (flt-1)),
non-receptor protein tyrosine kinases, for example LCK, Src, Frk, Btk, Csk,
Abl,
Zap70, Fes/Fps, Fak, Jak, Ack, and LIMK, growth factor receptor tyrosine
kinases
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29
such as VEGF-R2, FGF-R, TEK, Akt kinases, Aurora kinases (Aurora A, Aurora B,
Aurora C) and the like.
The compounds of Formulas I and Z can be inhibitors of protein kinases such
as, for example, the inhibitors of the checkpoint kinases such as Chkl, Chk2
and the
like. Preferred compounds can exhibit IC50 values of less than about 51rm,
preferably
about 0.001 to about 1.0 pm, and more preferably about 0.001 to about 0.1
/gym. The
assay methods are described in the Examples set forth below.
The compounds of Formulas I and Z can be useful in the therapy of
proliferative diseases such as cancer, autoimmune diseases, viral diseases,
fungal
diseases, neurological/neurodegenerative disorders, arthritis, inflammation,
anti-
proliferative (e.g., ocular retinopathy), neuronal, alopecia and
cardiovascular disease.
Many of these diseases and disorders are listed in U.S. 6,413,974 cited
earlier,
incorporated by reference herein.
More specifically, the compounds of Formulas I and Z can be useful in the
treatment of a variety of cancers, including (but not limited to) the
following:
tumor of the bladder, breast (including BRCA-mutated breast cancer,
colorectal,
colon, kidney, liver, lung, small cell lung cancer, non-small cell lung
cancer, head and
neck, esophagus, bladder, gall bladder, ovary, pancreas, stomach, cervix,
thyroid,
prostate, and skin, including squamous cell carcinoma;
leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell
lymphoma, T- cell lymphoma, Hodgkins lymphoma, non-Hodgkins lymphoma, hairy
cell lymphoma, mantle cell lymphoma, myeloma and Burkett's lymphoma;
chronic lymphocytic leukemia ("CLL");
acute and chronic myelogenous leukemia, myelodysplastic syndrome and
promyelocytic leukemia;
fibrosarcoma, rhabdomyosarcoma;
head and neck, mantle cell lymphoma, myeloma;
astrocytoma, neuroblastoma, glioma, glioblastoma, malignant glial tumors,
astrocytoma, hepatocellular carcinoma, gastrointestinal stromal tumors
("GIST") and
schwannomas;
melanoma, multiple myeloma, seminoma, teratocarcinoma, osteosarcoma,
xenoderoma pigmentosum, keratoctanthoma, thyroid follicular cancer and
Kaposi's
sarcoma.
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Due to the key role of kinases in the regulation of cellular proliferation in
general, inhibitors could act as reversible cytostatic agents which may be
useful in the
treatment of any disease process which features abnormal cellular
proliferation, e.g.,
benign prostate hyperplasia, familial adenomatosis polyposis, neuro-
fibromatosis,
5 atherosclerosis, pulmonary fibrosis, arthritis, psoriasis,
glomerulonephritis, restenosis
following angioplasty or vascular surgery, hypertrophic scar formation,
inflammatory
bowel disease, transplantation rejection, endotoxic shock, and fungal
infections.
Compounds of Formulas I and Z may also be useful in the treatment of
Alzheimer's disease, as suggested by the recent finding that CDK5 is involved
in the
10 phosphorylation of tau protein (J. Biochem, (1995)117, 741-749).
Compounds of Formulas I and Z may induce or inhibit apoptosis. The
apoptotic response is aberrant in a variety of human diseases. Compounds of
Formula I, as modulators of apoptosis, will be useful in the treatment of
cancer
(including but not limited to those types mentioned hereinabove), viral
infections
15 (including but not limited to herpevirus, poxvirus, Epstein- Barr virus,
Sindbis virus and
adenovirus), prevention of AIDS development in HIV-infected individuals,
autoimmune
diseases (including but not limited to systemic lupus, erythematosus,
autoimmune
mediated glomerulonephritis, rheumatoid arthritis, psoriasis, inflammatory
bowel
disease, and autoimmune diabetes mellitus), neurodegenerative disorders
(including
20 but not limited to Alzheimer's disease, AIDS-related dementia, Parkinson's
disease,
amyotrophic lateral sclerosis, retinitis pigmentosa, spinal muscular atrophy
and
cerebellar degeneration), myelodysplastic syndromes, aplastic anemia, ischemic
injury associated with myocardial infarctions, stroke and reperfusion injury,
arrhythmia,
atherosclerosis, toxin-induced or alcohol related liver diseases,
hematological
25 diseases (including but not limited to chronic anemia and aplastic anemia),
degenerative diseases of the musculoskeletal system (including but not limited
to
osteoporosis and arthritis) aspirin-sensitive rhinosinusitis, cystic fibrosis,
multiple
sclerosis, kidney diseases and cancer pain.
Compounds of Formula I, as inhibitors of kinases, can modulate the level of
30 cellular RNA and DNA synthesis. These agents would therefore be useful in
the
treatment of viral infections (including but not limited to HIV, human
papilloma virus,
herpesvirus, poxvirus, Epstein-Barr virus, Sindbis virus and adenovirus).
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Compounds of Formulas I and Z may also be useful in the chemoprevention of
cancer. Chemoprevention is defined as inhibiting the development of invasive
cancer
by either blocking the initiating mutagenic event or by blocking the
progression of pre-
malignant cells that have already suffered an insult or inhibiting tumor
relapse.
Compounds of Formulas I and Z may also be useful in inhibiting tumor
angiogenesis and metastasis.
Compounds of Formulas I and Z may also act as inhibitors of cyclin dependent
kinases and other protein kinases, e.g., protein kinase C, her2, raf 1, MEK1,
MAP
kinase, EGF receptor, PDGF receptor, IGF receptor, P13 kinase, weel kinase,
Src,
Abl and thus be effective in the treatment of diseases associated with other
protein
kinases.
Another aspect of this invention is a method of treating a mammal (e.g.,
human) having a disease or condition associated with kinases (e.g., CDKs, CHK
and
Aurora kinases) by administering a therapeutically effective amount of at
least one
compound of Formula I, or a pharmaceutically acceptable salt, solvate, ester
or
prodrug of said compound to the mammal.
A preferred dosage is about 0.001 to 1000 mg/kg of body weight/day of the
compound of Formula I. An especially preferred dosage is about 0.01 to 25
mg/kg of
body weight/day of a compound of Formula I, or a pharmaceutically acceptable
salt,
solvate, ester or prodrug of said compound.
The compounds of this invention may also be useful in combination
(administered.
together or sequentially) with one or more of anti-cancer treatments such as
radiation
therapy, and/or one or more anti-cancer agents different from the compound of
Formula I. The compounds of the present invention can be present in the same
dosage unit as the anti-cancer agent or in separate dosage units.
Another aspect of the present invention is a method of treating one or more
diseases associated with a kinase (such as CDK, CHK and Aurora), comprising
administering to a mammal in need of such treatment: an amount of a first
compound,
which is a compound of Formula 1, or a pharmaceutically acceptable satt,
solvate,
ester or prodrug thereof; and an amount of at least one second compound, the
second
compound being an anti-cancer agent different from the compound of Formula 1,
wherein the amounts of the first compound and the second compound result in a
therapeutic effect.
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Non-limiting examples of suitable anti-cancer agent is selected from the group
consisting of a cytostatic agent, cisplatin, doxorubicin, liposomal
doxorubicin (e.g.,
Caely) , Myocet , Doxil ), taxotere, taxol, etoposide, irinotecan, camptostar,
topotecan, paclitaxel, docetaxel, epothilones, tamoxifen, 5-fluorouracil,
methoxtrexate,
temozolomide, cyclophosphamide, SCH 66336, R115777 , L778,123 , BMS 214662 ,
Iressa , Tarceva , antibodies to EGFR, antibodies to IGFR (including, for
example,
those published in US 2005/0136063 published June 23, 2005), KSP inhibitors
(such
as, for example, those published in WO 2006/098962 and WO 2006/098961;
ispinesib, SB-743921.from Cytokinetics), centrosome associated protein E
("CENP-
E") inhibitors (e.g., GSK-923295), Gleevec , intron, ara-C, adriamycin,
cytoxan,
gemcitabine, Uracil mustard, Chlormethine, Ifosfamide, Melphalan,
Chlorambucil,
Pipobroman, Triethylenemelamine, Triethylenethiophosphoramine, Busulfan,
Carmustine, Lomustine, Streptozocin, Dacarbazine, Floxuridine, Cytarabine,
6-Mercaptopurine, 6-Thioguanine, Fludarabine phosphate, oxaliplatin,
leucovirin,
ELOXATINTM, Pentostatine, Vinblastine, Vincristine, Vindesine, Bleomycin,
Dactinomycin, Daunorubicin, Doxorubicin, Epirubicin, Idarubicin, Mithramycin,
Deoxycoformycin, Mitomycin-C, L-Asparaginase, Teniposide 17a-Ethinylestradiol,
Diethylstilbestrol, Testosterone, Prednisone, Fluoxymesterone, Dromostanolone
propionate, Testolactone, Megestrolacetate, Methyl prednisolo ne,
Methyltestosterone,
Prednisolone, Triamcinolone, Chlorotrianisene, Hydroxyprogesterone,
Aminoglutethimide, Estramustine, Medroxyprogesteroneacetate, Leuprolide,
Flutamide, Toremifene, goserelin, Cisplatin, Carboplatin, Hydroxyurea,
Amsacrine,
Procarbazine, Mitotane, Mitoxantrone, Levamisole, Navelbene, Anastrazole,
Letrazole, Capecitabine, Reloxafine, Droloxafine, Hexamethylmelamine, Avastin,
herceptin, Bexxar, bortezomib ("Velcade"), Zevalin, Trisenox, Xeloda,
Vinorelbine,
Porfimer, Erbitux, Liposomal, Thiotepa, Altretamine, Melphalan, Trastuzumab,
Lerozole, Fulvestrant, Exemestane, Fulvestrant, Ifosfomide, Rituximab, C225 ,
satriplatin, mylotarg, Avastin, Rituxan, panitubimab, Sutent, sorafinib,
Sprycel
(dastinib), nilotinib, Tykerb (lapatinib) and Campath.
If formulated as a fixed dose, such combination products employ the
compounds of this invention within the dosage range described herein and the
other
pharmaceutically active agent or treatment within its dosage range. For
example, the
CDC2 inhibitor olomucine has been found to act synergistically with known
cytotoxic
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33
agents in inducing apoptosis (J. Cell Sci., (1995) 108, 2897. Compounds of
Formulas I
and Z may also be administered sequentially with known anticancer or cytotoxic
agents when a combination formulation is inappropriate. The invention is not
limited in
the sequence of administration; compounds of Formulas I and Z may be
administered
either prior to or after administration of the known anticancer or cytotoxic
agent. For
example, the cytotoxic activity of the cyclin-dependent kinase inhibitor
flavopiridol is
affected by the sequence of administration with anticancer agents. Cancer
Research,
(1997) 57, 3375. Such techniques are within the skills of persons skilled in
the art as
well as attending physicians.
Accordingly, in an aspect, this invention includes combinations comprising an
amount of at least one compound of Formula I, or a pharmaceutically acceptable
salt,
solvate, ester or prodrug thereof, and an amount of one or more anti-cancer
treatments and anti-cancer agents listed above wherein the amounts of the
compounds/ treatments result in desired therapeutic effect.
Another aspect of the present invention is a method of inhibiting one or more
Aurora kinases in a patient in need thereof, comprising administering to the
patient a
therapeutically effective amount of at least one compound of Formula 1 or a
pharmaceutically acceptable salt, solvate, ester or prodrug thereof.
Another aspect of the present invention is a method of treating, or slowing
the
progression of, a disease associated with one or more Aurora kinases in a
patient in
need thereof, comprising administering a therapeutically effective amount of
at least
one compound of Formula 1 or a pharmaceutically acceptable salt, solvate,
ester or
prodrug thereof.
Yet another aspect of the present invention is a method of treating one or
more
diseases associated with Aurora kinase, comprising administering to a mammal
in
need of such treatment an amount of a first compound, which is a compound of
Formula 1, or a pharmaceutically acceptable salt, solvate, ester or prodrug
thereof;
and an amount of at least one second compound, the second compound being an
anti-cancer agent, wherein the amounts of the first compound and the second
compound result in a therapeutic effect.
Another aspect of the present invention is a method of treating, or slowing
the
progression of, a disease associated with one or more Aurora kinases in a
patient in
need thereof, comprising administering a therapeutically effective amount of a
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34
pharmaceutical composition comprising in combination at least one
pharmaceutically
acceptable carrier and at least one compound according to Formula 1, or a
pharmaceutically acceptable salt, solvate, ester or prodrug thereof.
In the above methods, the Aurora kinase to be inhibited can be Aurora A,
Aurora B and/or Aurora C.
Another aspect of the present invention is a method of inhibiting one or more
Checkpoint kinases in a patient in need thereof, comprising administering to
the
patient a therapeutically effective amount of at least one compound of formula
1 or a
pharmaceutically acceptable salt, solvate, ester or prodrug thereof.
Another aspect of the present invention is a method of treating, or slowing
the
progression of, a disease associated with one or more Checkpoint kinases in a
patient
in need thereof, comprising administering a therapeutically effective amount
of at least
one compound of formula 1 or a pharmaceutically acceptable salt, solvate,
ester or
prodrug thereof.
Yet another aspect of the present invention is a method of treating one or
more
diseases associated with Checkpoint kinase, comprising administering to a
mammal
in need of such treatment an amount of a first compound, which is a compound
of
formula 1, or a pharmaceutically acceptable salt, solvate, ester or prodrug
thereof; and
an amount of at least one second compound, the second compound being an anti-
cancer agent, wherein the amounts of the first compound and the second
compound
result in a therapeutic effect.
Another aspect of the present invention is a method of treating, or slowing
the
progression of, a disease associated with one or more Checkpoint kinases in a
patient
in need thereof, comprising administering a therapeutically effective amount
of a
pharmaceutical composition comprising in combination at least one
pharmaceutically
acceptable carrier and at least one compound according to formula 1, or a
pharmaceutically acceptable salt, solvate, ester or prodrug thereof.
In the above methods, the checkpoint kinase to be inhibited can be Chk1
and/or Chk2.
Another aspect of the present invention is a method of inhibiting one or more
cyclin dependent kinases in a patient in need thereof, comprising
administering to the
patient a therapeutically effective amount of at least one compound of formula
1 or a
pharmaceutically acceptable salt, solvate, ester or prodrug thereof.
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Another aspect of the present invention is a method of treating, or slowing
the
progression of, a disease associated with one or more cyclin dependent kinases
in a
patient in need thereof, comprising administering a therapeutically effective
amount of
at least one compound of formula 1 or a pharmaceutically acceptable salt,
solvate,
5 ester or prodrug thereof.
Yet another aspect of the present invention is a method of treating one or
more
diseases associated with cyclin dependent kinase, comprising administering to
a
mammal in need of such treatment an amount of a first compound, which is a
compound of formula 1, or a pharmaceutically acceptable salt, solvate, ester
or
10 prodrug thereof; and an amount of at least one second compound, the second
compound being an anti-cancer agent, wherein the amounts of the first compound
and
the second compound result in a therapeutic effect.
Another aspect of the present invention is a method of treating, or slowing
the
progression of, a disease associated with one or more cyclin dependent kinases
in a
15 patient in need thereof, comprising administering a therapeutically
effective amount of
a pharmaceutical composition comprising in combination at least one
pharmaceutically acceptable carrier and at least one compound according to
formula
1, or a pharmaceutically acceptable salt, solvate, ester or prodrug thereof.
In the above methods, the checkpoint kinase to be inhibited can be CDK1
20 and/or CDK2.
Another aspect of the present invention is a method of inhibiting one or more
tyrosine kinases in a patient in need thereof, comprising administering to the
patient a
therapeutically effective amount of at least one compound of Formula 1 or a
pharmaceutically acceptable salt, solvate, ester or prodrug thereof.
25 Yet another aspect of the present invention is a method of treating, or
slowing
the progression of, a disease associated with one or more tyrosine kinases in
a
patient in need thereof, comprising administering a therapeutically effective
amount of
at least one compound of Formula 1 or a pharmaceutically acceptable salt,
solvate,
ester or prodrug thereof.
30 Another aspect of the present invention is a method of treating one or more
diseases associated with tyrosine kinase, comprising administering to a mammal
in
need of such treatment an amount of a first compound, which is a compound of
Formula 1, or a pharmaceutically acceptable salt, solvate, ester or prodrug
thereof;
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36
and an amount of at least one second compound, the second compound being an
anti-cancer agent, wherein the amounts of the first compound and the second
compound result in a therapeutic effect.
Another aspect of the present invention is a method of treating, or slowing
the
progression of, a disease associated with one or more tyrosine kinases in a
patient in
need thereof, comprising administering a therapeutically effective amount of a
pharmaceutical composition comprising in combination at least one
pharmaceutically
acceptable carrier and at least one compound according to Formula 1 or a
pharmaceutically acceptable salt, solvate, ester or prodrug thereof.
In the above methods, the tyrosine kinase can be VEGFR (VEGF-R2), EGFR,
HER2, SRC, JAK and/or TEK.
Another aspect of the present invention is a method of inhibiting one or more
Pim-1 kinases in a patient in need thereof, comprising administering to the
patient a
therapeutically effective amount of at least one compound of Formula 1 or a
pharmaceutically acceptable salt, solvate, ester or prodrug thereof.
Yet another aspect of the present invention is a method of treating, or
slowing
the progression of, a disease associated with one or more Pim-1 kinases in a
patient
in need thereof, comprising administering a therapeutically effective amount
of at least
one compound of Formula 1 or a pharmaceutically acceptable salt, solvate,
ester or
prodrug thereof.
Another aspect of the present invention is a method of treating one or more
diseases associated with. Pim-1 kinase, comprising administering to a mammal
in
need of such treatment an amount of a first compound, which is a compound of
Formula 1, or a pharmaceutically acceptable salt, solvate, ester or prodrug
thereof;
and an amount of at least one second compound, the second compound being an
anti-cancer agent, wherein the amounts of the first compound and the second
compound result in a therapeutic effect.
Another aspect of the present invention is a method of treating, or slowing
the
progression of, a disease associated with one or more Pim-1 kinases in a
patient in
need thereof, comprising administering a therapeutically effective amount of a
pharmaceutical composition comprising in combination at least one
pharmaceutically
acceptable carrier and at least one compound according to Formula 1 or a
pharmaceutically acceptable salt, solvate, ester or prodrug thereof.
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37
The pharmacological properties of the compounds of this invention may be
confirmed by a number of pharmacological assays. The exemplified
pharmacological
assays which are described herein below have been carried out with compounds
according to the invention and their salts, solvates, esters or prodrugs.
This invention is also directed to pharmaceutical compositions which comprise
at least one compound of Formula I, or a pharmaceutically acceptable salt,
solvate,
ester or prodrug of said compound and at least one pharmaceutically acceptable
carrier.
For preparing pharmaceutical compositions from the compounds described by
this invention, inert, pharmaceutically acceptable carriers can be either
solid or liquid.
Solid form preparations include powders, tablets, dispersible granules,
capsules,
cachets and suppositories. The powders and tablets may be comprised of from
about
5 to about 95 percent active ingredient. Suitable solid carriers are known in
the art,
e.g., magnesium carbonate, magnesium stearate, talc, sugar or lactose.
Tablets,
powders, cachets and capsules can be used as solid dosage forms suitable for
oral
administration. Examples of pharmaceutically acceptable carriers and methods
of
manufacture for various compositions may be found in A. Gennaro (ed.),
Remington's
Pharmaceutical Sciences, 18th Edition, (1990), Mack Publishing Co., Easton,
Pennsylvania.
Liquid form preparations include solutions, suspensions and emulsions. As an
example may be mentioned water or water-propylene glycol solutions for
parenteral
injection or addition of sweeteners and opacifiers for oral solutions,
suspensions and
emulsions. Liquid form preparations may also include solutions for intranasal
administration.
Aerosol preparations suitable for inhalation may include solutions and solids
in
powder form, which may be in combination with a pharmaceutically acceptable
carrier,
such as an inert compressed gas, e.g. nitrogen.
Also included are solid form preparations that are intended to be converted,
shortly before use, to liquid form preparations for either oral or parenteral
administration. Such liquid forms include solutions, suspensions and
emulsions.
The compounds of the invention may also be deliverable transdermally. The
transdermal compositions can take the form of creams, lotions, aerosols and/or
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38
emulsions and can be included in a transdermal patch of the matrix or
reservoir type
as are conventional in the art for this purpose.
The compounds of this invention may also be delivered subcutaneously.
Preferably the compound is administered orally or intravenously.
Also contemplated are delivery methods that are combinations of the above-
noted delivery methods, Such methods are typically decided by those skilled in
the art.
Preferably, the pharmaceutical preparation is in a unit dosage form. In such
form, the preparation is subdivided into suitably sized unit doses containing
appropriate quantities of the active component, e.g., an effective amount to
achieve
the desired purpose.
The quantity of active compound in a unit dose of preparation may be varied or
adjusted from about 1 mg to about 100 mg, preferably from about 1 mg to about
50
mg, more preferably from about 1 mg to about 25 mg, according to the
particular
application.
The actual dosage employed may be varied depending upon the requirements
of the patient and the severity of the condition being treated. Determination
of the
proper dosage regimen for a particular situation is within the skill of the
art. For
convenience, the total daily dosage may be divided and administered in
portions
during the day as required.
The amount and frequency of administration of the compounds of the invention
and/or the pharmaceutically acceptable salts thereof will be regulated
according to the
judgment of the attending clinician considering such factors as age, condition
and size
of the patient as well as severity of the symptoms being treated. A typical
recommended daily dosage regimen for oral administration can range from about
1
mg/day to about 500 mg/day, preferably 1 mg/day to 200 mg/day, in two to four
divided doses.
Another aspect of this invention is a kit comprising a therapeutically
effective
amount of at least one compound of Formula I, or a pharmaceutically acceptable
salt,
solvate, ester or prodrug of said compound and a pharmaceutically acceptable
carrier,
vehicle or diluent.
Yet another aspect of this invention is a kit comprising an amount of at least
one compound of Formula I, or a pharmaceutically acceptable salt, solvate,
ester or
prodrug of said compound and an amount of at least one anticancer therapy
and/or
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39
anti-cancer agent listed above, wherein the amounts of the two or more
ingredients
result in desired therapeutic effect.
The invention disclosed herein is exemplified by the following preparations
and
examples which should not be construed to limit the scope of the disclosure.
Alternative mechanistic pathways and analogous structures will be apparent to
those
skilled in the art.
Where NMR data are presented, 1 H spectra were obtained on either a Varian
VXR-200 (200 MHz, 1 H), Varian Gemini-300 (300 MHz) or XL-400 (400 MHz) and
are
reported as ppm down field from Me4Si with number of protons, multiplicities,
and
coupling constants in Hertz indicated parenthetically. Where LC/MS data are
presented, analyses was performed using an Applied Biosystems API-100 mass
spectrometer and Shimadzu SCL-1 OA LC column: Altech platinum C18, 3 micron,
33mm x 7mm ID; gradient flow: 0 min - 10% CH3CN, 5 min - 95% CH3CN, 7 min -
95% CH3CN, 7.5 min - 10% CH3CN, 9 min - stop. The retention time and observed
parent ion are given.
The following solvents and reagents may be referred to by their abbreviations
in parenthesis:
Thin layer chromatography: TLC
dichloromethane: CH2CI2
dimethylformamide: DMF
1,1'-Bis(diphenylphosphino)ferocene : dppf
dithiothreitol: DTT
ethyl acetate: AcOEt or EtOAc
methanol: MeOH
N-iodosuccinimide : NIS
trifluoroacetate: TFA
triethylamine: Et3N or TEA
butoxycarbonyl: n-Boc or Boc
nuclear magnetic resonance spectroscopy: NMR
liquid chromatography mass spectrometry: LCMS
high resolution mass spectrometry: HRMS
milliliters: mL
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millimoles: mmol
microliters: l
grams: g
milligrams: mg
5 room temperature or rt (ambient): about 25 C.
dimethoxyethane: DME
The synthesis of the inventive compounds is illustrated below. Also, it should
be noted
that the disclosure of commonly-owned U.S. 6,919,341 and U.S. Appl. No.
11/598186
is incorporated herein by reference.
10 EXAMPLE 1
Br 1.25 NIS, DMF I
N
equiv NaSMe ~N^ 'Br 60 C, 95% NBr
N YN McOH, rt, 95% N~ /N Part B NN
Part A SN SN
cat. Pd(PPh3)4, K2CO3
Part C trimethylboroxine, DMF
100 C, 90%
N NIS, DMF N^
/N 7
N 60 C, 95% N N
SN Part D SN
Part A: The title compound was prepared according to US20060106023 (Al).
Part B: To a solution of compound from Example 1, Part A (2.00 g, 8.19 mmol)
in DMF
(50 ml-) was added N-iodosuccinimide (1.84 g, 8.19 mmol). The reaction mixture
was
15 stirred at 60C for 16 hours. The mixture was cooled to 25C and
concentrated. The
residue was dissolved in DCM with a small amount of methanol and then loaded
on
the column. Purification by column chromatography (Si02, 40% ethyl
acetate/hexanes) afforded compound as a white solid 2.30 g (76%). 'H-NMR (400
MHz, DMSO-d6) d 8.3 (s, 1 H), 7.8 (s, 1 H), 2.6 (s, 3H). HPLC-MS tR = 1.87 Min
(UV
20 254nm)= Mass calculated for formula C7H5BrIN3S 370.01, observed LC/MS m/z
370.9
(M+H).
Part C: A suspension of bromide from Part A (45.6 g), Pd(PPh3)4 (10.8 g),
potassium
carbonate (77.4 g), trimethylboroxine (46.9 g) and potassium carbonate (77.4
g) in
DMF (410 mL) was heated overnight under nitrogen at 105C. After cooling, the
25 mixture was diluted with ethyl acetate (1 L), washed with brine (2 x 500
mL), dried
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41
(magnesium sulfate), filtered, concentrated and purified by chromatography on
silica
gel. The title compound was obtained as a pale yellow solid (21.4 g, 64%).
Part D: To a DMF (400 mL) solution of compound from Example 1, Part C (21.8 g)
was added N-iodosuccinimide (26.9 g) and the resulting mixture was heated
overnight
at 60C. The mixture was concentrated and water (400 mL) was added. After
stirring 1
hr at rt, saturated sodium carbonate was added (250 mL) and subsequently
stirred an
additional 30 min at rt. The mixture was filtered, washed with water, methanol
(100
mL) and the filter cake was dried overnight under vacuum. A brown solid was
obtained
(31.4 g, 87%).
EXAMPLE 2
H3C CH3
01-Bu
H3C t0
H3C O-B - O
1
BrN + rN Bra
N
N N Ot-Bu N N
SCH3 0 SCH3
A solution of tert-butyl 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-
pyrazol-1-yl)acetate (318 mg, 1.03 mmol) in 1,4-dioxane (3 mL) and water (0.30
mL)
was added to an argon degassed mixture of compound from Example 1, Part B (292
mg, 0.79 mmol), Pd(dppf)C12 (58 mg, 0.079 mmol) and potassium phosphate (503
mg,
2.37 mmol). The reaction was heated to 40 C and allowed to stir for 12 hours.
The
reaction was cooled to room temperature, filtered through Celite eluting with
ethyl
acetate then concentrated to dryness. Purification of the crude residue by
flash
chromatography (Si02, 12 g; 5% to 40% ethyl acetate in hexanes) afforded the
title
compound as a light brown solid 244 mg (73%). 1H NMR (300 MHz, CDCI3) 6 7.94
(s,
1 H), 7.81 (s, 1 H), 7.80 (s, 1 H), 7.65 (s, 1 H), 4.94 (s, 2H), 2.68 (s, 3H),
1.51 (s, 9H).
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42
EXAMPLE 3
H
N'NN S` N
O!-Bu O
N- N"'~
0 Bra N CH3
I' \
Br1~\ N N =HCl
NY _N S "q/ N
SCH3
CH3
3-Chloroperoxybenzoic acid (204 mg, 1.18 mmol) was added to a room
temperature solution of ester from Example 2 (244 mg, 0.58 mmol) in methylene
chloride (3 mL). The reaction was allowed to stir for 1 hour. Upon completion,
the
reaction was concentrated to dryness then taken up into ethyl acetate (50 mL).
The
solution was washed with saturated aqueous sodium bicarbonate solution (50 mL)
and brine (2 x 50 mL) then dried (sodium sulfate), filtered and concentrated
to dryness
to give 220 mg of crude sulfoxide. The crude sulfoxide (220 mg, 0.48 mmol) in
dimethylsulfoxide (2.5 mL) was added to a premixed solution of sodium hydride
(106
mg, 1.45 mmol) and 2-amino-4-methylisothiazole (78 mg, 0.68 mmol) in
dimethylsulfoxide (2.5 mL). The reaction was stirred for 20 minutes then
quenched
with saturated aqueous ammonium chloride (50 mL). The aqueous layer was
extracted with diethyl ether (2 x 50 ml-) and ethyl acetate (2 x 50 mL). The
combined
organic layers were washed with brine (2 x 50 mL), dried (sodium sulfate),
filtered and
concentrated to dryness. Purification of the resultant residue by flash
chromatography
(Si02, 12 g; 5% to 40 % ethyl acetate in methylene chloride) and then on prep-
HPLC
afforded the title compound as a yellow solid 2 mg (0.8%). 1H NMR (300 MHz,
CDCI3)
J 8.35 (s, 1 H), 8.32 (s, 1 H), 8.02 (s, 1 H), 7.93 (s, 1 H), 7.17 (s, 1 H),
6.90 (s, 1 H), 5.37
(s, 2H), 2.63 (s, 3H), 2.46 (s, 3H). HPLC tR = 4.62 min (UV 254nm). Mass
calculated for
formula C19H16BrN9OS2 529.01; observed MH+ (APCI MS) 531.1 (m/z).
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43
EXAMPLE 4
IOH
N- N
0
H3C N
H3C N N 1''
N1~ N
=HCI
SEMN I `N HNC
'
N
NQ N
A solution of fert-butyl 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1
H-
pyrazol-1-yl)acetate (135 mg, 0.44 mmol) in 1,4-dioxane (3 ml-) and water
(0.30 ml-)
was added to a nitrogen flushed mixture of iodide (200 mg, 0.34 mmol),
Pd(dppf)CI2
(25 mg, 0.034 mmol) and potassium phosphate (216 mg, 1.02 mmol). The reaction
mixture was heated to 90 C and allowed to stir for 12 hours. Upon completion,
the
reaction was allowed to cool to room temperature and then was concentrated to
dryness. Purification of the resultant residue by flash chromatography (Si02;
12 g;
10% to 80% ethyl acetate in methylene chloride) afforded the desired coupled
intermediate. Trifluoroacetic acid (1 ml-) was added to a room temperature
solution of
the desired coupled ester (80 mg, 0.125 mmol) in methylene chloride (3 mL).
The
reaction was stirred for 12 hours then concentrated to dryness. Purification
of the
resultant residue by prep-HPLC afforded the title compound as a yellow solid
40 mg
(45%). 'H NMR (300 MHz, CD30D) d 8.34 (s, 1 H), 8.26 (s, 1 H), 8.08 (s, 1 H),
8.01 (s,
1 H), 7.36 (s, 1 H), 5.15 (s, 2H), 4.43 (s, 2H), 3.17-2.99 (m, 2H), 2.71-2.55
(m, 2H),
2.64 (s, 3H), 2.06-1.71 (m, 5H), 1.65-1.46 (m, 1 H). HPLC tR = 3.82 min (UV
254nm).
Mass calculated for formula C21H24N802S 452.17; observed MH+ (ESI MS) 453.1
(mlz).
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44 .
EXAMPLE 5
OH
N,NII-I
0
H3CY I- N\\ H3C
N N
N
IIIN
--- NY N
HCl
SEM'N S`N HN S.
I r CH3 IN JH3
N
N
Example 5 was prepared in a similar manner to Example 4. 'H NMR (300
MHz, CD3OD) d 8.34 (s, 1 H), 8.26 (s, 1 H), 8.08 (s, 1 H), 8.01 (s, 1 H), 7.36
(s, 1 H),
5.15 (s, 2H), 4.43 (s, 2H), 3.17-2.99 (m, 2H), 2.71-2.55 (m, 2H), 2.64 (s,
3H), 2.06-
1.71 (m, 5H), 1.65-1.46 (m, 1 H). HPLC to = 3.82 min (UV 2541m). Mass
calculated for
formula C21H24N802S 452.17; observed MH' (ESI MS) 453.1 (m/z).
EXAMPLE 6
The compounds shown in column 2 of Table I were prepared as follows:
OH
H
N 0 N,N",-1
H3CN 0
l`` H3C~N
NY _N =TFA T'
IIN N \' N =HCI
S ~
IN HN
IN
N
R
R
A mixture of acid (1 equivalent), the respective amine (1.5 equivalents), HATU
(1.5 equivalents), and diisopropylethylamine (3 equivalents) in anhydrous DMF
(500
pL) was stirred at room temperature for 2 hours. The reaction was then
concentrated
under reduced pressure, purified by preparative HPLC and conversion to the
hydrochloride salt afforded compounds shown in Column 2 of Table 1.
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Table 1
MS
Example Column 2 MW MH+ HPLC
m/z tR
H
N N
N- N
r 11
O
H3C
6-1 N N 556.2 557.4 3.51
N
HN S,N CH3
N
/ l
H N
N`N"-~ N
r
0
H3C
6-2 N \ N 556.2 557.2 3.48
N
IN CH3
HN
N
N
H
N-N~
r '1
0
H3C //
6-3 N \ N 556.2 557.3 3.48
N
HN
TL SN
r N 3
CH
N
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46
H
N- N"'~ N
CF3
0
H3CN
6-4 623.2 624.6 5.34
Nom/ N
IHN S'
N CH3
N
/ ICF3
N-N~(
N
'11
0
6-5 H3C N \ N \ 623.2 624.6 5.35
N
HN '
N CH3
N
iD/CF3
H
N
N- N~
i I`
0
H3C-T5;'~'N 623.2 624.5 5.39
6-6
NY-,-- N
HN
CH
IN 3
N
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47
H
N,NI",T N
r 0 OCH3
H3C //
6-7 N \ NN 585.2 586.9 4.96
r N CH3
HN
N
/ I
H
N,N'-"~ N OCH3
0
H3C
N
6-8
N 585.2 586.3 4.85
Y `N
IHN
N 3
CCH
N
OCH3
H
N,N~
0
H3C
6-9 ~N 585.2 586.2 4.82
N
HN
IN CH3
N
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48
H
N N,CH3
N` N) CH3
3
H3C
6-10 N Y-,-- 598.3 599.8 3.71
N
HN S
I N CH3
N
H3CO
H N,NN 16
i
O
H3CN
6-11 N 571.2 572.4 5.12
HN s
IN 3
N
CN OCH3
N,
i
~ O
H3C
N
6-12 N N 571.2 572.4 4.98
xN
I 'N CH3
N
H
N`N"- tV I
ci 0 OCH3
H3C\
1" N ~
6-13 N N 571.2 572.5 4.87
HN
I /N CH3
N
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49
H
N` N ^ NYI N
O
H3C
6-14 N\')'-N 542.2 543.7 3.93
HNIE s
N CH3
N
H
N-
NN N
O
H3C
N\
6-15 N ),)'-N 542.2 543.6 3.60
HN
N CH3
N
H
N\NN I
0 II N
H3C`r N
6-16 N L N 542.2 543.6 3.67
L IN CH3
HN
N
H
N`N N
F
0
H3C)~ N \ \ I
6-17 N\~-N 559.6 260.2 5.10
Euv
/N
No
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WO 2009/097233 PCT/US2009/031972
H
N,NN
N F
H3C
6-18 N\ N 559.6 560.3 5.13
HN
"N
No
N
N. Nom(
111
0 /
H3C
N F F
6-19 N \L N 577.6 578.9 5.26
HN
IN
No
H
N,N'-~ N I
0 F
H3C\ ~N F
~/ \
6-20 N N 563.6 564.7 5.43
HN"
IN
No
H
0
N,N"-~ N
F
H3C N F
6-21 N\),- 563.6 564.7 5.34
HN
IN
No
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51
H
N-Nl~( N
Ill
0 Nr 0
H3CN
6-22 N \ N 546.6 547.6 3.96
xN
r N 3
N
CH N zl-_"OH
N- NN
11
0
H3C
578.7 579.8 3.39
6-23 Y-,-- N
NN
HN
CH3
/NN
N
^/OCH3
1(`N
N-NN
II
0
H3C
6-24 NN 592.7 593.9 3.52
N Y'N
HN
IN JH3
N
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52
H3C
NN=CH3
N, N , N
i 11
0
6-25 H3CYN 605.8 606.8 3.35
N~/ N
IHN S
N 3
N
CH C
Cs
N- N
i 11
0
H3C\ ~N6-26 N J` 551.7 552.8 4.47
N~/'N
HN
I N 3
CH
N
N
N N
OH
0
H3C //
6-27 N \ N 571.7 572.5 4.59
N
HN
I N CH3
N
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53
OH
N-N~(
i 11
0
H3C
6-28 N \ 571.7 572,4 4.26
NY'N
HN .
IN J H3
N
^^N,CH3
N-N1N,
i 1-
0
6-29 H3C N 548.7 549.8 3.45
N
IN H3
N
N I \
0
i
0
H3C-fl~ N
6-30 N ,N 545.6 546.5 4.53
HN ,
i N 3
C
N
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54
H I
N-N')( S
i1
0
H3C ^ N
6-31 N -N 561.7 562.1 4.72
HN S..
IN CH3
N
H
N-N~
r
O /
H3C N F
6-32 N \~-- 545.2 546.5 5.08
HN
SN N
H
N,N'-f N I F
r
0 /
H3C N \ F
6-33 NY-,--N 563.2 564.7 5.32
HNC
SN N
~J
N
N,N~ F
0 F
H3C /
6-34 N 577.2 578.9 4.97
HN
SSN
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NHZ
N,Nom(
r
0
H3C{N
6-35 N 451.1 452.2 3.44
H N T ,
rN /~
N )
H
N` N/\(N-CH3
r 11
O
H3C
N
6-36 N \~ N 465.2 466.7 3.57
Huv
IN
No
H ^ ,CH3
N-N~N '~
r / 0 CH3
H3C N
6-37 N)-- 'N 521.2 522.9 4.71
I-IN
1IN
No
CH3
N- N N,CH3
r 1`
0
H3C-r~ N
6-38 NY N 479.2 480.1 3.73
HN s
IN
No
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56
0
-(NJ
N-N 11
r 0
H3C
6-39 N 521.2 522.9 3.73
N T\~ N
HN
No
H
N- N'-~ N--/'OH
0
H3C N
6-40 N lN 495.2 496.9 3.42
T
HN
tNQ_____
~~/0
N- N 1
r HN
~ NOZ
H3C\ N \ ~ 1
6-41 N.I```N 600.2 601.7 4.91
HN S
CH3
IN
N
N, N
HN
H3C
N
6-42 N N NO2 600.2 601.6 4.90
xN s
/N CH3
N
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57
N, N10
HN
H3C\ ~N \ / I
N~ N
600.2 601.6 4.90
6-43 HN s NO,
rN
N
CH3
F /
H
N
N' Cl
O
H3C N 6-44 N Y-1-- N 607.2 608.5 5.12
HN ,
N
N
CH3
F
N- Nom( N / I
I1 Cl
O
H3C\ N
6-45 NiN 607.2 608.4 5.23
HN s
rN
N
CH3
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58
F
N
Cl
<:JN'1
0
H3C ~N
~ 1
6-46 Ny `- N 607.2 609.4 5.23
HN ,
IN
N
CH3
C1
H
N,N"Nf N
0 F
H3C N
6-47 NY N 607.2 609.4 5.27
HN
IN
'
N
CH3
N,N"Y N
0 C1
H3C T\ N
NY N 589.2 591.8 5.15
6-48
IN
N
CH3
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59
_",a
H
N,N,~,N Cl
O
H3C N
6-49 Nyl~ N 589.2 591.8 5.16
IAN S
~N
N
CH3
C1
H
N- N
O
H3C
` N
6-50 NYt_`N 589.2 591.8 5.17
HN
IN
N
CH3
H
N
CH3
N`N O
H3CY;~ N
6-51 N 569.3 570.7 5.01
N
HN,J~ 'N
N
CH3
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N
N' N O CH3
H3C
N
6-52 N 569.3 570.7 4.99
1' N
IIN
N
N
CH3
H
j:)
N
N' N-1-1
0
H3CN
1/ \
6-53 N1'\ N 555.3 556.3 4.82
HN
IN
N
CH3
N'N^/N p
i n
O
H3C\ ^N \ CF3
6-54 N7`\ N 609.6 610.4 5.54
HN
I N 3
~SH
N
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61
N,N~N I
0 CF3
H3C\ N
T/ \
6-55 N \) N 609.6 610.6 5.59
HN
/N CH3
N
F
H
N`N")~ N
0
H3C\ N F
6-56 Ny 577.6 578.8 5.17
HN
C
/N 3
N
F
N`N O
N 6 F
6-57 H3C N
N N 577.6 578.9 5.03
HN S
i,N 3
N
CH CH3
N',,-'- N,
N` N C %
O H3
H3C\T^ / N
6-58 N N 536.7 537.7 3.45
UN
N JH3
N
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62
N\I,'-N'CH3
N-N'~ H
0
H3CN
6-59 N 522.6 523.9 3.43
N
HN S,
IN CH3
N
OH
N-Nl~-(
~ I11
0
6-60 H3C N N 549.6 550.7 3.76
N
HN
IN 3
CH F
N
I
H
N-N'N
F
O
6-61 H3C\
' N 577.22 578.9 5.20
NYl-- N
HN S'
IN
N )
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F
N- N )(N F
1I
0
H3C
6-62 Y-- NN 577.22 578.9 5.25
NN
HN
IN
No
F
H
N-N" f
11
0
H3C
6-63 NN 559.23 560.2 5.12
NY N
HN
I IN
No
v F
N`N--~ N
H3C N
6-64 N N 545.21 546.5 5.17
xN
N
No
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64
H
N`N I
O F
H3C
N
Nlrl~
6-65 N\,J,-N 545.21 546.4 5.21
I-IN
IN
N
F
F
N-N"-(N
O
H3CN
6-66 N \~ N 577.6 578.9 5.14
FI
IN
%
N
F
F b
H N` NX--~ N
0
6-67 H3C~N 591.6 592.8 5.03
N)/'N
HN S.
N
N
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H
NN~N
K0
H3C N
6-68 N~ N 559.6 560.4 5.00
HN
I IN
N
H
N,N~(N I ~~ F
0
H3CN
6-69 N N 559.6 560.4 5.11
HN
IN
N
N
N,N
0
H3C //
6-70 N\ N 569.7 570.6 5.05
N
HN S
L IN
N
N,NN
/
0
H3C
6-71 NY `N 555.6 556.4 4.96
HNI S.
N
N
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F
H
N-NN
i
O FI
H3C ^N
6-72 NT ,N 577.6 578.3 4.85
HN S,
1N CH3
N
H
N-NN
'II
O
H3CN HO
6-73 N Ti- N 571.6 572.4 4.33
HN S
C
N CH3
N
H N-
N F
N, N I ~
11
0
H3C)5;^ N
6-74 N N 560.6 561.3 4.92
i
HN S,N CH3
N
F
N,N----( N bl-
0
6-75 H3C N
N ,N 560.6 561.2 4.52
yl-
HHN,11 S'
i N CH3
N
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F
N,N--~ N
r ,N
0
H3C
N
6-76 N N 560.6 561.2 3.75
HN,I[ S%
1 N 3
N
CH
N.NN I N
r 111
O F
H3C N
6-77 N. 560.6 561.3 4.53
T N
HN S
LiN JH3
N
H
N`N")~ N
0
H3CN (N l
6-78 N N N 653.8 654.8 3.63
HN CH3
,N CH3
N
H
N,N~N
0
H3C)/ N \
6-79 N. (t N 541.6 542.6 4.91
T
HN' S
L iN CH3
N
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H
~JN(Q
0
H3C\^N cNl
6-80 N N o 640.8 641.6 3.84
HN S%
I N CH3
N
H".Z)s
N`N^ N
0
H3CN
6-81 TNN 519.6 520.4 4.42
HN ,
iN CH3
N
H
N\N~N,,~
0 N
H3C`^
~/ N
548.6 549.6 4.44
6-82 N\ N
HN
C
L iN 3
N
HO
H
N .N~N I
0 ,N
H3C~N
6-83 N . 558.6 559.4 3.79
HN
L/N .CH3
N
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69
H Cl
N`N-- ,N I
0
H3C\ N 6-84 Ny-N 576.1 576.8 5.41
HN S'
iN CH3
N
H
N,N'-(N 1
11011 Cl
H3C N
6-85 N N 576.1 576.8 5.37
HN,,C S,
1 N CHI
N
H
N
N.N 11 , 0INHZ
O S
H3C
~ 0
N
6-86 N 1 N 620.7 621.7 4.28
HN,IL S,
,N 3
CH H
N
N,N'-( N `,N
0
H3C`..N OH
6-87 NY 'N 572.6 573.3 3.77
HN,I[ S
1N 3
C
N
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s
N
II
N6N"
0
6-88 H3C N 561.7 562.3 4.71
NY N
HNI S%
L i N H3
N
O
H 1I CH
N ~/~ ' 3
N-Nl'-~ H
0
H3C` 7 N \
6-89 N 536.6 537.6 3.69
HN S,
iN H3
N
H
N-N--~ NY CH3
0 N-N
H3C\ ^N
6-90 N N 563.7 564.9 4.24
HN %
i!(:: 3
CCH H
N
N`N '-~ N
'OH
0
H3Cy NI_ N
6-91 N l 563.7 564.8 3.89
HN,I~ S,N 3
C
N
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H
0
cJN"cO
H3C_ " N
6-92 N N 549.6 550.8 3.98
HN SIN CH3
N
H
N
iN 01 QN
H3C N
6-93= N N 558.6 559.5 3.72
HN SN CH3
N
H
11 N
~\N
0 F_
H3C\ ^N
6-94 N 1- N 546.6 547.7 3.52
HN S%
~N
No
H 02N
N _
NN~
0 N
6-95 H3C` ^N
N 573.6 574.5 4.47
N
HN SI N
N
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H NH
N
H3C` ^N
6-96 N. N 566.6 567.7 4.63
HN S,
iN H3
N
H
O N /
H3C\ ^N
6-97 N 592.7 593.9 4.16
T N
HN S
C
,N H3
N
N,N"I(N I i
IO N
H3C\
1" N \
6-98 N N 592.7 593.9 3.84
HN S,
iN H3
N
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73
EXAMPLE 7
F
-~[ F
F F N~N~
F F N/ 1 N N N '/Z
0
H_b S
Br/ 0 Pan A Part B N N
BPin
iS
F F
N F
N`N ON I/ F N O l i
Part C Part D
N N N N
0=i=0 It, NE
Part A: To a solution of 4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)-1 H-
pyrazole
(2.2 g, 11.32) and bromide (2.83 g, 11.32 mmol) in DMA (5 mL) was added
potassium
carbonate (1.9 g, 13.6 mmol). The mixture was heated at 60C for 20 hr. To the
reaction mixture was added half sat'd ammonium chloride and ethyl acetate. The
organic phase was washed with water (2x), brine and dried (sodium sulfate).
Concentration and purification by chromatography (50% ethyl acetate in
hexanes)
afforded the title compound as a pale yellow solid. (2.1 g, 51 %). 1H NMR (300
MHz,
DMSO-d6) 610.3 (1 H, br s), 7.95 (1 H, s), 7.7-7.6 (1 H, app t), 7.59 (1 H,
s), 7.1-7.2
(1 H, m), 5.12 (s, 2H), 1.23 (s, 12H).
Part B: A mixture of Example 1, Part D (1.49 g), boronate from Example 7, Part
A
(2.13 g), PdCl2(dppf) (0.398 g), potassium phosphate (2.07 g), in DME (45 mL)
and
water (5 mL) was heated at 95C overnight. The reaction was allowed to cool,
diluted
with ethyl acetate and filtered through Celite. The filtrate was washed with
water, brine
and dried (sodium sulfate). Chromatography afforded the title compound.
Part C: A solution of the compound from Example 7, Part B (260 mg) in THE (25
mL)
at rt was added MCPBA (288 mg) in one portion. After 1 hr at rt, ethyl acetate
was
added and was washed with sat. sodium bicarbonate (2x), brine and dried
(sodium
sulfate). Concentration afforded the title compound that was used without
further
purification.
Part D: A solution of the compound from Example 7, Part C (1 equiv), amine (5
equiv),
DIEA (5 equiv) in NMP was heated at 50C overnight. The reaction mixture was
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74
concentrated and purified by Prep-LC. Using this general procedure compounds
listed
in Table 2 were prepared.
Table 2
MS
Example Column 2 MW MH+ HPLC
m/z tR
F
H F
N- N" N I
0
7-1 H3C N 383.35 384.1 3.01
NY_N
NHZ
N F
0
7-2 H3C N 397.38 398.2 2.59
y
N\~ N
/NH
N &F
0
7-3 H3C 427.4 428.2 2.57
N
N\~ N
HO^~ NH
F
F
_NN
/ II
0
7-4 H3CN 482.53 483.3 2.90
NY N
fN~iNH
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F
H
F
0
7-5 H3 N 496.56 497.3 2.83
Y
NY N
NH
F
H F
N,NN
0
7-6 H3CNN 496.51 497.2 3.02
N\ L N
NH
OJ
WK,
7-7 425.43 426.1 3.00
"-~N
N)--`N
NH
F
N`N ^ /N F
So'
7-8 H3C 510.54 511.1 2.38
~N \
ON, Ny 'N
~ NH
N-NN
0
7-9 496.56 497.2 2.71
NY `N
HN ~
N_
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76
H F
N
O
7-10 N 542.58 543.2 3.22
NY-,-- N
HN
F
H F
N-N 0
7-11 N 518.52 519.2 2.39
NN/. N
HN~O \ N
H F
N\NN
0
7-12 N \ N N 547.56 548.2 4.29
N O
HN ~
O,
F
H
N`to
F
0
7-13 ~Nl\ 488.49 489.1 2.44
NN
HN ~,,~,I
N
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77
N F
0
7-14 N 499.51 500.2 3.36
N rj4trN
HN
H F
N`N ^ ,N
7-15 ~N \ 469.49 470.2 3.09
NN
HN
OH
F
F
N-Nl"~ N
7-16 ~N \ 457.43 458.2 2.95
NrJ::zN
HN
HO OH
H F
N-,, ,-N
0
7-17 N -N 504.49 505.2 2.31
HN
OH
N
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78
F
N`Nll~(N
O
N
7-18 N - 504.49 505.1 2.60
HN
HO INS
H F
F
N
0
N
7-19 N, -N 504.49 505.1 2.37
HN
HO
F
H F
N-N-,vN
0
~N \
7-20 N ~N 504.49 505.1 2.15
HN
HO N
N-,, F
0
7-21 N \ N N 474.47 475.1 2.56
HN
/ I
N
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79
F
N`N,~ N
7-22 NN 549.2 550.2 3.12
N/ N
HIN S
N N
N`N~N F
7-23 N \ N 563.2 564,2 3.21
N
HNYS
N
F
N`N')~ N
0
7-24 ~N 558.2 559.2 1.42
NY N
HN 0
I~ NJ
N`N ^ /N F
N
7-25 N N 482.53 483.3 2.36
HN
N
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EXAMPLE 8
OH N-
N- N,N --
o CH
3 0
H3CN H C.N CI H3CN
11N3 RN H,
~N =ZTFA H3C CH3 N =2HCI
HN $ HN 5
I /N CH3 I iN CH3
N~ N
To a stirring suspension of carboxylic acid (1 equiv) in dichioromethane at 0
C was
5 added 1-chloro-N,N-2-trimethyl-1-propenylamine (5 equiv). After stirring for
1 hour the
amine was added as a solution in dichioromethane or pyridine. When the
reaction
was deemed complete by HPLC analysis the mixture was concentrated under
reduced pressure. Purification by prep-HPLC and conversion to the
hydrochloride salt
provided the compounds listed in Table 3.
Table 3
MS
Example Column 2 MW MH+ HPLC
tR
m/z
H CI
N\
N
O
H3C
N
8-1 N N 577.1 587.0 4.64
HN SN
L /N CH
N
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81
H
N-N"'Yk /
IOI N`N
H3CN
8-2 N N 549.6 550.9 4.11
HN S,
1N CH3
N
H N
N-N-(NY
II
0 N
H3C\
~"/ N
8-3 N11 'N 543.6 544.6 4.15
HN SN
L /N 3
N
CH
N.N"I1I a,,,,
(N O N OH
H3C 7
8-4 N 'L N 558.6 559.4 3.73
1T
HN gN
L/N CH3
N
H
N,N"~ N N
O
H3C\ 'N \
8-5 N N 543.6 544.5 6.99
HN,I[ ~
~N CH3
N
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82
H CH3
N`N CIN
H3C
N
8-6 N. 556.6 557.3 3.73
HN
L /N 3
N
CEXAMPLE 9
N H ~
N- N 1
N~ 1 / N. N /
F F F H3C\
T/^ N + LAH H3C N F
NNN -21M THF NY 'N =2HCI
RN S HN S
TL /N
CH3
CH3 IN N V
N
NV
Lithium aluminum hydride (2 mg, 2.2 equiv) was added to a stirring suspension
of amide (14 mg, 1 equiv) in THE (1 mL) at room temperature. HPLC analysis
after 15
minutes showed no starting material so reaction was quenched with methanol,
concentrated under reduced pressure, purified by prep-HPLC, and conversion to
the
hydrochloride salt afforded the title compound as a white solid 6.5 mg (47%)).
1H
NMR (300 MHz, DMSO-d5) d 12.3 (s, 1 H), 10.1 (bs, 1 H), 8.41 (s, 1 H), 8.03
(s, 1 H),
7.89 (m, 2H), 7.30 (s, 1 H), 6.97 (m, 1 H), 6.55 (m, 2H), 4.39 (m, 4H), 3.67
(m, 2H),
3.39 (m, 2H), 2.80 (m, 2H), 2.48 (s, 3H), 1.79 (m, 4H), 1.05 (m, 1 H), 0.89
(m, 3H).
HPLC tR = 5.58 min (UV 254nm). Mass calculated for formula C28H31F2N9S 563.24;
observed MH+ (ESI MS) 564.8 (mlz).
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83
EXAMPLE 10
ooo
O-B Part A 0-B Part B O-B
H N N
Br N3
NH2
N,N,--J
Part C,D H3C N
NI-,-- N
SEM N S
N
No
Part A: Potassium hydroxide (1.45 g, 10.0 equiv) was added to a stirring
solution of 4-
(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)-1 H-pyrazole (500 mg, 1.00
equiv) in
DMSO (5 mL) at room temperature. After stirring for 1 hour, 1,2-dibromoethane
(9.69
g, 20.0 equiv) was added. The reaction was stirred for 16 hours at which time
TLC
indicated no starting material remained so the reaction was quenched with
water (10
ml-) and extracted with ethyl acetate (3 x 15 mL). The combined organics were
washed with brine (20 mL), dried over sodium sulfate, filtered, concentrated
under
reduced pressure, and purification by silica gel chromatography (12g Si02,
dichloromethane to 5% methanol in dichloromethane) afforded the desired
boronate
as a yellow oil 350 mg (45%).
Part B: A mixture of the boronate from example 10 part A (250 mg, 1,00 equiv),
sodium azide (108 mg, 2.00 equiv), and sodium iodide (124 mg, 1.00 equiv) in
DMSO
(2 ml-) was stirred at 50 C for 2 hours at which time TLC indicated no
starting
material remained. The reaction was allowed to cool to room temperature, then
quenched with water (8 ml-) and extracted with ethyl acetate (3 x 15 mL). The
combined organics were washed with brine (15 mL), dried over sodium sulfate,
filtered, concentrated under reduced pressure, and purification by silica gel
chromatography (12g Si02, dichloromethane to 5% methanol in dichloromethane)
afforded the desired boronate as a yellow solid 170 mg (78%).
Part C: To a mixture of aryl iodide from Part B (315 mg, 1.00 equiv),
PdCI2(dppf) (39
mg, 0.10 equiv), and potassium phosphate (228 mg, 2.00 equiv) under nitrogen
was
added a solution of the boronate from Part B (170 mg, 1.20 equiv) in 1,4-
dioxane (1.5
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84
mL), followed by water (0.15 mL). The mixture was stirred at 90 C for 17
hours at
which time TLC indicated no starting material remained. The reaction was
allowed to
cool to room then diluted with ethyl acetate (8 mL) and washed with brine (10
mL),
dried over sodium sulfate, filtered, concentrated under reduced pressure, and
purification by silica gel chromatography (12g Si02, dichioromethane to 5%
methanol
in dichioromethane) afforded the desired azide as a brown solid 125 mg (39%).
Part D: To a stirring solution of azide from Part C (125 mg, 1.00 equiv) in
1,4-dioxane
(2 mL) and water (0.2 mL) was added poly-styrene bound triphenylphosphine (84
mg,
1.20 equiv). The reaction was stirred at room temperature for 3 days at which
time
TLC indicated no starting material remained. The mixture was filtered, the
mother
liquor concentrated under reduced pressure, and the resulting residue purified
by
silica gel chromatography (12 g Si02, dichioromethane to 10% methanol in
dichloromethane) affording the desired amine as a brown oil 77 mg (64%).
EXAMPLE 11
N
N- ~~NHZ N .. N F
N N 0
H3C~N H3C I \ NN
NY 'N NY N =2HCI
SEM'N S HIV S~
TL/N I IN
No NQ
N-Methyl morpholine (14 mg, 2.00 equiv) was added to a stirring mixture of
carboxylic acid (14 mg, 1.50 equiv) and HATU (39 mg, 1.50 equiv) in DMF (0.5
mL).
After 30 minutes, the amine from example 10 (38.5 mg, 1.00 equiv) was added as
a
solution in DMF (0.5 mL). The mixture was stirred for 5 hours at which time
HPLC
indicated no starting material remained. The mixture was concentrated and the
residue dissolved in 1,4-dioxane (1 mL). A solution on HCI in dioxane (1 mL,
4M in
dioxane) was added and the mixture was sonicated for 1.5 hours at which time
HPLC
indicated no starting material remained. The mixture was concentrated under
reduced
pressure, purified by prep-HPLC, and conversion to the HCI salt afforded the
desired
compound as a yellow solid 6 mg (14%). 'H NMR (300 MHz, CD30D) d 8.90 (m, 1H),
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8.72 (m, 1 H), 8,36 (s, 1 H), 8.24 (s, 1 H), 8.01(m, 3H), 7.32 (s, 1 H), 4.58
(m, 2H), 4.42
(s, 2H), 3.94 (m, 2H), 3.60 (m, 2H), 3.09 (m, 2H), 2.61 (s, 3H), 1.81 (m, 6H),
1.55 (m,
2H). HPLC tR = 3.97 min (UV 254m). Mass calculated for formula C27H29FN10OS
560.22; observed MH+ (ESI MS) 561.3 (m/z).
5
EXAMPLE 12
H F
NH, N
N- F
N N Q
H3CN 4 H3CN
NY 'N NY 'N =2HCI .00 SEM.N S HN S.
IN I IN
N~ NQ
To a stirring solution of amine from example 10 (38.5 mg, 1.00 equiv) and
triethylamine (14 mg, 2.00 equiv) in dichloromethane (0.75 mL) was added 2,3-
10 difluorobenzoylchloride (13 mg, 1.10 equiv) dropwise. HPLC analysis after 4
hours
showed no starting material so reaction was quenched with saturated aqueous
sodium bicarbonate (2 mL) and then extracted with dichloromethane (3 x 1 mL).
The
combined organics were concentrated and the resulting residue was dissolved in
1,4-
dioxane (1 mL) and a solution on HCI in dioxane (1 mL, 4M in dioxane) was
added
15 and the mixture was sonicated for 1.5 hours at which time HPLC indicated no
starting
material remained. The mixture was concentrated under reduced pressure,
purified
by prep-HPLC, and conversion to the HCI salt afforded the desired compound as
a
yellow solid 5 mg (11 %). 'H NMR (300 MHz, CD30D) d 8.34 (s, 1 H), 8.21 (s, 1
H),
8.06 (s, 1 H), 7.92 (s, 1 H), 7.44 (m, 2H), 7.36 (2, 1 H), 7.20 (m, 1 H), 4.56
(m, 2H), 4.41
20 (s, 2H), 3.92 (m, 2H), 3.59 (m, 2H), 3.08 (m, 2H), 2.59 (s, 3H), 1.78 (m,
6H). HPLC tR
= 4.61 min (UV 254nm). ass calculated for formula C28H29F2N9OS 577.22;
observed
MH+ (ESI MS) 578.8 (m/z).
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86
EXAMPLE 13
H `
~(N
N`N \\ \ i N
0
F
H3CIN H3CN
N'/ N --T NN =2HC1
SEM'N TS HN S
iN
CHO IN N RI
R,
Sodium triacetoxyborohydride (1.50 equiv) was added to a stirring mixture of
aldehyde
(1.00 equiv), amine (1.20 equiv), and acetic acid (1.00 equiv) in 1,2-
dichloroethane at
room temperature. The mixture was stirred until no starting material remained
as
judged by TLC. The reaction was then quenched with 1 N NaOH and extracted
three
times with chloroform. The combined organics were dried over sodium sulfate,
filtered, and concentrated. The residue was then added as a solution in
dioxane (1
equiv) to a mixture of boronate (1.50 equiv), PdCI2(dppf) (0.10 equiv), and
potassium
phosphate (2.00 equiv) under nitrogen. Water was added and the mixture was
stirred
at 90 C for 17 hours at which time HPLC indicated no starting material
remained.
The reaction was allowed to cool to room temperature the diluted with ethyl
acetate,
washed with water, dried over sodium sulfate, filtered, concentrated under
reduced
pressure, and purification by silica gel chromatography afforded the coupled
product.
This material was dissolved in 1,4-dioxane, HCI (4N in dioxane) was added and
the
mixture was sonicated until such time that HPLC indicated no starting material
remained. The mixture was concentrated under reduced pressure, purified by
prep-
HPLC, and conversion to the hydrochloride salt afforded the title compounds as
white
solids in Table 4.
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Table 4
MS
Example Column 2 MW MH+ HPLC
m/z tR
H
N`N'' N
I N
0 F
'H3C -rN
13-1 N \ - 560.6 561
N .4 3.75
HN S
N ;CH3
No
H
N
N-NI"f
0 F 1
H3C HIV
13-2 N 560.6 561.3 3.78
HN S
L /N CH3
N
H
N' NN
N~ I ~N
0 F%
H3C N
13-3 N TN 574.6 575.6 3.97
HN .
/ N H3C CH3
N
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H
N Q
N~. I /N
''011 F
H3C
N
13-4 N N 560.6 561.4 3.76
HN S,
tNO_____
H
N, Iq I
0 F
H3C\ N
13-5 N 1" N 576.6 578.2 3.63
HN S, OCH3
L /N
N
H
N, N
N^/
i n N
0 F
~N
13-6 3 N ~N 576.6 578.2 3.62
Y'L HN S
I /,N /OCH3
No
H
~(
N-N 11
O ~F?
F
H3C\^N
13-7 N N ,2HCI 593.7 594.7 5.13
HN S ~,N /OCH3
N I
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H
N
N`N 1 /
O
F
F
H;CN
13-8 N` ~N =2110 593.7 594.7 5.15
HN S. OCHJ
i N
N
EXAMPLE 14
F F F
F 5 NHZ Part A F N CI Part B F I NCI
NH Z H N
SEM
HJC CH3
HJC O
H3C C=B F
Part C -C\.
,' N/ F
~N
H
Part A: Chloroacetic acid (5.11 g, 1.3 equiv) was added to a stirring solution
of 2-
amino-3,4-difluoroaniline (6.00 g, 1 equiv) in 6 N hydrochloric acid (28 mL).
After
stirring for 18 hours at 95 C the reaction was cooled to room temperature,
made
basic with 10% aqueous potassium carbonate and extracted with ethyl acetate
(850
mL). The organic layer was separated, dried over sodium sulfate, filtered,
concentrated under reduced pressure, and purification by silica gel
chromatography
(120 g Si02, hexanes to 60% ethyl acetate in hexanes) afforded the desired
benzimidazole as a pink solid 6.24 g (74%).
Part B: A mixture of the benzimidazole from example 14 part A (4.18 g, 1.00
equiv),
potassium carbonate (8.53 g, 3.00 equiv) in DMF (50 ml-) was stirred at room
temperature for 5 minutes at which time 2-(trimethylsilyl)ethoxymethyl
chloride (4.0
mL, 1.1 equiv) was added. After stirring at room temperature for 18 hours the
reaction
was quenched with a saturated aqueous solution of sodium bicarbonate (40 ml-)
and
concentrated under reduced pressure to a residue. The residue was diluted with
ethyl
acetate (500 mL) and washed with water (150 mL), dried over sodium sulfate,
filtered,
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concentrated under reduced pressure, and purification by silica gel
chromatography
(120 g Si02, hexanes to 50% of ethyl acetate in hexanes) afforded the desired
benzimidazole as a brown oil 3.11 g (45%).
Part C: To a solution of 4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)-1 H-
pyrazole
5 (1.65 g, 1 equiv), benzimidazole from Example 14 part B (3.11 g, 1.1 equiv)
in DMA
(57 ml-) was added potassium carbonate (3.51 g, 3 equiv). The mixture was
heated at
50 C for 18 hours. The reaction mixture was poured into water (250 mL),
extracted
with ethyl acetate (500 mL), the organic layer washed with brine (250 mL),
dried over
sodium sulfate, filtered, concentrated under reduced pressure, and
purification by
10 silica gel chromatography (80 g Si02, hexanes to 50% of ethyl acetate in
hexanes)
afforded the desired boronate as a off-white solid 2.67 g (64%).
EXAMPLE 15
F
~
N- N ~ \ F
' HNN
H3C~N~ HjC` N
N N - NY N -2HC1
SEM N S HN S
rN I rN
Rt ~ Rt
N N
R, RZ
15 To a mixture of aryl iodide (100 mg, 1.00 equiv), PdCI2(dppf) (12 mg, 0.10
equiv), and
potassium phosphate (71 mg, 2.00 equiv) under nitrogen was added a solution of
the
boronate from Example 14 part C (123 mg, 1.20 equiv) in 1,4-dioxane (2.0 mL),
followed by water (0.2 mL). The mixture was stirred at 90 C for 18 hours at
which
time TLC indicated no starting material remained. The reaction was allowed to
cool to
20 room temperature, then diluted with ethyl acetate (100 mL) and washed with
water
(30mL), dried over sodium sulfate, filtered, concentrated under reduced
pressure, and
purification by silica gel chromatography (12 g Si02, dichloromethane to 10%
methanol in dichloromethane) afforded the desired coupled product. This
material
was dissolved in 1,4-dioxane, HCI (4N in dioxane) was added and the mixture
was
25 sonicated until such time that HPLC indicated no starting material
remained. The
mixture was concentrated under reduced pressure, purified by prep-HPLC, and
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conversion to the hydrochloride salt afforded the title compounds as off-white
solids in
Table 5.
Table 5
MS
Example Column 2 MW MH+ HPLC
m/z tR
N,N~ N F
F
ICI
H3C N \ -
NY '
15-1 N 560.2 561.2 4.74
HN s,
IN
V
F
N N
HN F
H3CNN
INY N
15-2 HN s 574.2 575.7 4.99
TIN
0
EXAMPLE 16
F
N N F
N
N
F13CN~ H3C~N SEM
NN N
N
SEM'N I S/N SEM I
N S
N
CO:CH3 CO CH
2 3
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To a mixture of aryl iodide (2.15 g, 1.00 equiv), PdCI2(dppf) (288 mg, 0.10
equiv), and
potassium phosphate (1.67 mg, 2.00 equiv) under nitrogen was added a solution
of
the boronate from Example 14 part C (2.51 g, 1.20 equiv) in 1,4-dioxane (47
mL),
followed by water (4.7mL). The mixture was stirred at 90 C for 3 hours at
which time
TLC indicated no starting material remained. The reaction was allowed to cool
to
room then diluted with ethyl acetate (700 mL) and washed with water (250 mL),
dried
over sodium sulfate, filtered, concentrated under reduced pressure, and
purification by
silica gel chromatography (120 g Si02, hexanes to 100% ethyl acetate) afforded
the
desired coupled product as a brown foam 2.04 g (66%).
EXAMPLE 17
F F
N-N~ / \ F N`N \ F
N ~~ N
SEM N SEM
H3CN Part A H3CN
N\ lj=N N\l N
SEM N S` N S
iN SEM q/N
CO2CH3 F CHO
j 'N
N,N ~ ~ \ F
HN J
Part B H3CN
NN =2HC1
HN
I IN
Rt
N
RZ
Part A: To a stirring solution of the coupled product from Example 16 (2.04 g,
1 equiv)
in tetrahydrofuran (63 mL) at -78 C was added DIBAL-H (1 M in
dichioromethane, 6.5
mL, 2.5 equiv) dropwise. The mixture was stirred at -78 C for 5 hours at
which time
thin layer chromatography (30% ethyl acetate/hexanes) indicated the reaction
was
complete. The mixture was quickly poured into stirring saturated aqueous
sodium
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potassium tartrate and stirred at room temperature for 14 hours. The mixture
was
extracted with ethyl acetate (500 mL), the organic layer separated, dried over
sodium
sulfate, filtered, and concentrated under reduced pressure affording the
aldehyde as a
brown foam 1.96 g (100%).
Part B: Sodium triacetoxyborohydride (1.50 equiv) was added to a stirring
mixture of
aldehyde (1.00 equiv), amine (1.20 equiv), and acetic acid (1.00 equiv) in 1,2-
dichloroethane at room temperature. The mixture was stirred until no starting
material
remained as judged by TLC. The reaction was then quenched with 1 N NaOH and
extracted three times with chloroform. The combined organics were dried over
sodium sulfate, filtered, and concentrated. This material was dissolved in 1,4-
dioxane, HCI (4N in dioxane) was added and the mixture was sonicated until
such
time that HPLC indicated no starting material remained. The mixture was
concentrated under reduced pressure, purified by prep-HPLC, and conversion to
the
hydrochloride salt afforded the title compounds as off-white solids in Table
6.
Table 6
MS
Example Column 2 MW MH` HPLC
m/z tR
N,N^/N
HN
H3CN
NY _N
17-1 HN 538.6 539.8 4.08
IN
N
CH3
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F
N`N^ /N
~' F
HN
H3C N
NY
17-2 HN 588.23 589.1 4,84
N
N
CH3
CH3
F
N,N^/N
HN F
H3C N
NY IN
17-3 HN IN 604.2 605.9 4.35
N
HO
F
N,N^ N
F
HN
H3C ~
NN
HN S
17-4 I 'N 604.2 605.2 4.38
N
,O
H3C
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EXAMPLE 18
F
N F
N
N N \ F N, ~ F
N
SEM SEM
H3CN Part A H'C~N
Nom/ 'N NYN
.N S.
SEM I /N SEM'N S.
/N
CHO
F OH F
N N
N-N~~ F N N \ F
N HN i
SE 14
Part B H3Cy.N ( Part C H3CN
IN"- N IN 'N =2HCl
SEM.N S HN S
IN IN
RI
OMs N
R2
Part A: Sodium borohydride (2.0 equiv) was added to a stirring mixture of
aldehyde
from Example 17 part A (1.00 equiv) in acetic acid (5.7 ml-) in 1,2-
dichloromethane
5 (17 ml-) at room temperature. The mixture was stirred until no starting
material
remained as judged by TLC. The reaction was then quenched with 2N NaOH (11 ml-
)
and a saturated solution of aqueous sodium bicarbonate (35 mL). After stirring
at
room temperature for 15 minutes, the phases were separated and the organic
layer
was dried over sodium sulfate, filtered and concentrated under reduced
pressure to
10 afford the alcohol as a brown foam 1.01 g (100%).
Part B: Methane sulfonyl chloride (2 equiv) was added to a stirring solution
of alcohol
from Example 18 part A (1 equiv) and tnethylamine (4 equiv) in THE (40 mL).
After
stirring at room temperature for 30 minutes the reaction was quenched with a
saturated aqueous solution of ammonium chloride (14 ml-) and water (14 mL),
15 extracted with dichloromethane (2x80 mL), dried over sodium sulfate,
filtered and
concentrated under reduced pressure affording the mesylate as a brown foam
1.04 g
(90%).
Part C: To a stirring solution of mesylate from Example 18 part B (1 equiv),
amine (3
equiv), sodium iodide (0.5 equiv) in THE (1.0 ml-) was added
diisopropylethylamine (3
20 equiv) and the reaction heated at 60 C for 18 hours. The reaction was
cooled to
room temperature, diluted with dichloromethane (50 mL) and the organic layer
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96
washed with water (30 mL), brine (30 mL), dried over sodium sulfate, filtered
and
concentrated under reduced pressure. This material was dissolved in 1,4-
dioxane,
HCl (4N in dioxane) was added and the mixture was sonicated until such time
that
HPLC indicated no starting material remained. The mixture was concentrated
under
reduced pressure, purified by prep-HPLC, and conversion to the hydrochloride
salt
afforded the title compounds as off-white solids in Table 7.
Table 7
MS
Example Column 2 MW MH+ HPLC
m/z tR
N-N^
~ HN
H3CN
NY'N
18-1 HIV I S,N 606.2 607.1 4.95
N
~O
N`N^
~ HN
H3C N
TN/~
18-2 618.2 619.9 4.67
HN gIN `
OH
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N,N^ N
F
HN
H3C ~N
18-3 N N 562.2 563.5 4.48
HN S,
C/N
N
jJN'Th'H3CN
18-4 HN 592.2 593.5 4.37
/,N
N
OH
EXAMPLE 19
The following compounds (Table 8) can be prepared by a method similar to the
method described in Example 18.
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Table 8
Example Column 2
H
n ,N
N 60
I
H3CN F
19-1 NyN
11N
NQ
H
IN
~JN'O
F
H3C ~N
19-2 N
HN
No
H p
N~
N,N/~
NH
H3CcN ~;i F
19-3 N l/ ' N F
HN
No
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H
N,N/--~i
NH
H3CNI_ \ CF
19-4 N~N
HN s,
IN
No
H
N
N,N
O
H3CN F
19-5 N N F
HN
N
tNQ
H
N
N,N^i
O
H3C N F
19-6 Nj `N N
HN
N
No
N,1V N
HN
H3C\ N \
19-7 N N
HN g
N
No
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100
N-N^
H3CN \
19-8 N
1-IN s,
/N
No
N`N,---N,N
S
H3C~N \ F ~ \
19-9 NN N
HN S
/N
No
N-Nl'~'y N N
O
H3C N F
19-10 N N N
HN
It
No
N'NO,
N
N
H3C TN F
19-11 Nom/ N
HN s.
I /N
No
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H
N
N N 0
O F
H3C\ Nl` F
19-12 NN
xN
I ~N
No
H
NN~
- O F
H3C\~N
19-13 Nl N
HN
No
H
NN
N~ IO O I iN
F
H3CT~N
19-14 NY 'N
HN
NQ
H
N]o
S OF SIN
H3C~N
19-15 N T\ N
HN N
No
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0
S I N ~^~
F
H3C . N
T/ \
19-17 N
t N
HN
/N
No
H
S N)
0 N
F
H3C N
1I_~\
19-16 Nom/ N
H IN ,
L/N
No
H
N
S
N O F N
H3C N
19-17 N ,J N
FtN .
N
_N
ID
N
NN S 0
F
H3CI-r N
lI_~\
19-18 NY N
HN
/N
No
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H
N 0 F ~N
H3C N
19-19 NN
HN s,
N~
N H
0
H3CN F Jn/\
-,N
19-20 N L N
1-IN S,
/N
No
N N
H3C7N \ F ,-N
19-21 N
HNI S,
N
No
H
N
N\ S 0 F N
H3C~N
19-22 N
N
HN S
I /N
No
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104
N 0
HN N~NH
H3C ~ F
T'
19-23 N N
HN S/N
N0
N 0
HN`CN'-~NH
H3C\--N F
19-24 N
HN IN
No
N
HN'CN
H3C~N
N _ N ~ N N H
19-25 ~, I O-F
" /N
F
No
N
HN'CN
H3C\
N N~N ~ N/ NH
19-26 IY_
HN \
` /N
N
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EXAMPLE 20
H3CN H3C:r" N
INN N)--I-- N
SEM' N S SEM, N I S,
i CH3 /N H}
N
A solution of trimethylsilyl acetylene (46 mg, 0.46 mmol) in triethylamine
(0.5 mL) was
added to a nitrogen flushed mixture of iodide (136 mg, 0.23 mmol),
palladium(0)
triphenylphosphine (26 mg, 0.02 mmol) and copper(l) iodide (8.6 mg, 0.4 mmol).
The
reaction was stirred at room temperature for 12 hours then diluted with ethyl
acetate
(25 mL). The reaction contents were passed through Celite, concentrated, and
placed
onto a column (Si02; 12g; 10% to 50% ethyl acetate in hexanes) which afforded
the
desired coupled intermediate. The intermediate was dissolved in methanol (8
mL)
then treated with potassium carbonate (790 mg). The reaction was stirred at
room
temperature for 72 hours then diluted with methylene chloride (50 mL). The
organic
layer was washed with saturated aqueous sodium bicarbonate (50 mL), dried
(sodium
sulfate), filtered and concentrated to dryness. The resultant residue was
placed onto
a flash column (Si02; 4 g; 10% to 50% ethyl acetate in hexanes) to afford the
title
compound as a white solid 19 mg (17%). 'H NMR (300 MHz, CDCI3) 6 7.91 (s, 1
H),
7.82 (s, 1 H), 7.31 (s, 1 H), 6.67 (s, 2H), 3.91 (s, 1 H), 3.81 (t, J = 6.4
Hz, 2H), 3.71 (s,
2H), 3.03-2.87 (m, 2H), 2.66 (s, 3H), 2.19-1.99 (m, 2H), 1.85-1.63 (m, 5H),
1.01 (t, J =
6.4 Hz, 2H), 0.93 (d, J = 5.6 Hz, 3H), 0.00 (s, 9H).
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EXAMPLE 21
H F H F
Br N ,6F N ~ F
Sodium azide (116 mg,.1.79 mmol) was added to a solution of bromide (408 mg,
1.63
mmol) in dimethylformamide (30 mL). The reaction mixture was then heated to 60
C
and stirred for 12 hours. Upon completion, the reaction was cooled to room
temperature and the solvent was removed in vacuo. The resultant residue was
taken
into ethyl acetate (75 mL) then washed with sodium bicarbonate (50 mL), water
(50
mL) and brine (50 mL). The organic layer was then dried (sodium sulfate),
filtered,
and concentrated to dryness. The resultant solid was placed onto a flash
column
(Si02; 12 g; 10% to 50% ethyl acetate in hexanes) to give the title compound
as a
white solid 240 mg (69%). 'H NMR (300 MHz, CDCI3) 6 8.28 (br s, 1H), 8.12-7.96
(m, 1 H), 7.16-7.02 (m, 1 H), 7.01-6.87 (m, 1 H), 4.20 (s, 2H).
EXAMPLE 22
F
H F
N-N N 1
H'c N
II'' H;C
N N i -rN
N. ~N =2HCI
S E M N S` T
I ~N CH
3 ~ S
/~ I IN CH;
NI
N
Copper powder (5 mg, 0.08 mmol) was added to a solution of alkyne (19 mg, 0.04
mmol) from Example 21 and azide (16 mg, 0.08 mmol) from Example 22 in t-butyl
alcohol (0.3 mL) and water (0.6 mL). The reaction was stirred at room
temperature for
72 hours then diluted with ethyl acetate (100 mL). The organic layer was
washed with
water (100 mL) and brine (100 mL) then dried (sodium sulfate), filtered and
concentrated. The resultant residue was placed onto a flash column (Si02; 4 g;
0% to
10% methanol in methylene chloride) to afford the coupled intermediate. The
desired
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107
intermediate was then dissolved in dioxane (2 mL) and treated with 4 N HCI in
dioxane (2 mL). The reaction was sonicated at room temperature for 1 hour. The
solvent was removed and the residue was purified by prep-HPLC (95:5 to 5:95
water/acetonitrile with 0.1 % trifluoroacetic acid). The fractions were
collected and
dried and the residue treated with 0.2 N HCI and freeze-dried to afford the
title
compound as a white solid 6.9 mg (28%). 'H NMR (300 MHz, CD30D) d 8.65 (s, 1
H),
8.58 (s, 1 H), 8.13 (s, 1 H), 7.77-7.70 (m, 1 H), 7.21 (s, 1 H), 7.17-7.04 (m,
2H), 5.59 (s,
2H), 4.38 (s, 2H), 3.71-3.46 (m, 2H), 3.13-3.09 (m, 1 H), 2.79-2.63 (m, 1 H),
2.58 (s,
3H), 2.06-1.74 (m, 4H), 1.32-1.11 (m, 1 H), 1.00 (d, J = 6.4 Hz, 3H). HPLC tR
= 5.45
min (UV 254nm). Mass calculated for formula C27H28F2N10OS 578.2; observed MH+
(ESI MS) 579.8 (m/z).
EXAMPLE 23
"'N-'~ o
0
F13C~N~
H;C\/~N
N\\~N l
TT N+~N =2HCI
SEM N SkN
T H3
N
Example 24 was prepared in a similar manner to Example 15. 'H NMR (300 MHz,
CD30D) J 8.30 (s, 1 H), 8.21 (s, 1 H), 8.03 (s, 1 H), 8.00 (s, 1 H), 7.35 (s,
1 H), 5.31-5.22
(m, 1 H), 4.49-4.37 (m, 4H), 3.91 (s, 4H), 3.69-3.46 (m, 2H), 3.10-2.91 (m, 1
H), 2.80-
2.66 (m, 1 H), 2.63 (s, 3H), 2.07-1.74 (m. 4H), 1.39-1.12 (m, I H), 1.00 (d, J
= 6.5 Hz,
3H). HPLC tR = 7.36 min (UV 254nm). Mass calculated for formula C24H30N802S
494.2;
observed MH+ (ESI MS) 495.8 (m/z).
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108
EXAMPLE 24
N OH H
~N N
''011 N-N~ ~1NBoc
H3CN 0
NY IN H3CN
HN S
N CH3 HN S
N ) I N CH3
N
Example 25 was prepared in a similar manner to Example 6. 'H NMR (300 MHz,
CD30D) 6 8.20 (s, 1 H), 7.94 (s, 1 H), 7.85 (s, 1 H), 7.77 (s, 1 H), 7.18 (s,
1 H), 4.98 (s,
2H), 4.37 (br s, 2H), 4.17-3.81 (m, 2H), 3.71-3.40 (m, 2H), 3.20-3.05 (m, 1
H), 3.03-
2.84 (m, 2H), 2.79-2.62 (m, 1 H), 2.52 (s, 3H), 2.06-1.97 (m, 4H), 1.97-1.69
(m, 5H),
1.46 (s, 9H), 1.32-1.14 (m, 1 H), 1.00 (d, J = 6.4 Hz, 3H).
EXAMPLE 25
N-NN-C NBoc N,N~NNH
O , O
H3C-1~ N H3CN
N\l LN N` N =2HCI
FIN S HN S.
I A N CH3 /~CH3
N
Trifluoroacetic acid (2 mL) was added to a solution of amide (20 mg, 0.02
mmol) in
methylene chloride (2 mL). The reaction was allowed to stir at room
temperature for 2
hours. The solvent was removed and the resultant residue placed onto a prep-
HPLC
(95:5 to 40:60 water/acetonitrile with 0.1% trifluoroacetic acid). The
collected fractions
were concentrated then treated with 0.2 N HCI and freeze-dried to afford the
title
compound as a white solid 3.2 mg (23%). 1H NMR (300 MHz, CD30D) 6 8.23 (s, 1
H),
7.96 (s, 1 H), 7.91 (s, 1 H), 7.85 (s, 1 H), 7.21 (s, 1 H), 5.00 (s, 2H), 4.39
(br s, 2H), 4.09-
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3.93 (m, 1 H), 3.68-3.39 (m, 4H), 3.20-3.04 (m, 2H), 3.04-2.89 (m, 1 H), 2.79-
2.63 (m,
1 H), 2.51 (s, 3H), 2.24-2.09 (m, 2H), 2.05-1.67 (m, 6H), 1.32-1.12 (m, 1 H),
1.01 (d, J =
6.4 Hz, 3H). HPLC tR = 3.47 min (UV 254nm). Mass calculated for formula
C271"136N,o0S 548.3; observed MH+ (ESI MS) 549.9 (m/z).
EXAMPLE 26
I t
N_~ .LN NaBH4 N_ N Pd(PPh3)4
N'SEM AcOH/DCM \ N K3PO4, DMF
OHC S Part A / \" SEM Part B
N HO NS
~l D /I I o
F N~
F F H N,N F H F H N,N
N_
F Iv Part D
Part C
N NN
N N_`~N
7
\ N SEM N NH
' ---r
HO N'S OHC SEM N N_N'S
Part A: To a solution of iodide (390 mg, 0.757 mmol) in 10 mL of CH2CI2 was
added 8
mL of AcOH. NaBH4 (57 mg, 1.51 mmol) was then added in one portion. The
reaction was stirred at room temperature for 15 min. It was diluted with 100
mL of
CH2CI2, and neutralized by 5 N NaOH (aq.). To the mixture was added 100 ml of
saturated aqueous NaHCO3. The resulting mixture was stirred at room
temperature
for 30 min. The organic layer was isolated. It was dried over anhydrous
Na2SO4, and
then concentrated. The residue was purified by flash chromatography eluting
with
60% EtOAc/ CH2CI2 to give 390 mg of the title compound. 1H NMR (400 MHz,
CDCI3)
d 7.72 (s, 1 H), 7.60 (s, 1 H), 7.13 (s, 1 H), 6.58 (brs, 2H), 4.78 (s, 2H),
3.72 (t, 2H), 2.60
(s, 3H), 0.95 (t, 2H), -0.10 (s, 9H).
Part B: To a mixture of alcohol from Part A (80 mg, 0.15 mmol), boronate from
Example 7, Part A (84 mg, 0.23 mmol) and Pd(PPh3)4 (17.8 mg, 0.015 mmol) was
added 2 mL of DMF followed by 3 M aqueous K3PO4 solution (0.21 mL, 0.63 mmol).
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The reaction mixture was heated at 65 C for 18 h. It was diluted with 30 mL
of EtOAc
and washed with 1 N aqueous NH4CI solution (20 mL x 2). The organic layer was
concentrated under vacuum. The residue was purified by flash chromatography
eluting with 4% MeOH/CH2CI2 to give 68 m g of S. 'H NMR (400 MHz, CDCI3) 3
8.90
(brs, 1 H), 8.02-8.12 (m, 1 H), 8.00 (s, 1 H), 7,87 (s, 1 H), 7.68 (s, 1 H),
7.57 (s, 1 H), 7.15
(s, 1 H), 6.90-7.13 (m, 2H), 6.65 (brs, 2H), 5,09 (s, 2H), 4.75-4.81 (m, 2H),
3.77 (t, 2H),
2.80 (brs, 1 H), 2.60 (s, 3H), 0.95 (t, 2H), -0.10 (s, 9H).
Part C: To a solution of alcohol from Part B (31 mg, 0.049 mmol) in 1.5 mL of
THE,
was added 3 ^L of water followed by Dess-Martin periodinane (64 mg, 0.15
mmol).
The reaction mixture was stirred at room temperature for 30 min. It was
diluted with 5
mL of THF. The mixture was filtered. The filtrate was diluted with 20 mL of
CH2CIZ
and washed with 10 mL of saturated aqueous NaHCO3 solution. It was dried over
anhydrous Na2SO4 and then concentrated to give.30 mg of the title compound
which
was used in the subsequent reactions without further purification.
Part D: A solution of aldehyde (12 mg, 0.019 mmol), 3,3-dimethylpiperidine (22
mg,
0.19 mmol) in 1 mL of CH2CI2 was stirred at room temperature for 30 min. To
the
solution was added NaBH4 (3.6 mg, 0.096 mmol) followed by 0.3 mL of MeOH. The
reaction was stirred at room temperature for 1 h. It was diluted with 10 mL of
CH2CI2
and 10 mL of saturated aqueous NaHCO3 solution. The resulting mixture was
stirred
for 1 h. The organic was separated and concentrated under vacuum. The residue
was purified by flash chromatography eluting with NH4OH (aq.)/MeOH/ CH2CIZ
(1:10:190) to give 10 mg of the SEM-protected title compound. To a solution of
SEM-
protected material (10 mg, 0.014 mmol) in 2 mL of THF heated at 80 C was
added
0.2 mL of 4 N HCI in dioxane. The reaction was stirred at 80 C for 1.5 h. It
was
cooled to room temperature and then added 8 mL of ether. The solid was
collected by
filtration and washed with ether to give 8.7 mg of the title compound. 'H NMR
(400
MHz, CD30D) d 8.38 (s, 1 H), 8.15 (s, 1 H), 8.08 (s, 1 H), 8.02 (s, 1 H), 7.72-
7.81 (m,
1 H), 7.27 (s, 1 H), 7.03-7.20 (m, 2H), 5.30 (s, 2H), 4.35-4.60 (m, 2H), 3.55-
3.65 (d,
1 H), 3.00-3.10 (m, 1 H), 2.80-2.90 (d, 1 H), 2.60 (s, 3H), 1.81-2.10 (m, 2H),
1.40-1.69
(m, 2H), 1.20 (s, 3H), 1.00 (s, 3H). HPLC-MS tR = 2.96 min (UV 254nm)= Mass
calculated for formula C29H31F2N9OS 591.2; observed MH+ (LCMS) 592.3 (m/z).
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EXAMPLE 27
By essentially the same procedure set forth in Example 26, only replacing 3,3-
dimethylpiperidine with other respective amines in Part A, compounds shown in
column 2 of Table 9 were prepared.
TABLE 9
LCMS
Example Column 2 MW MH+ HPLC
m/z MS tR
J~
F NH l
F N,N
9-1 N \ N N 577.2 578.3 2.87
NH
N-S
.0
p
F NH'---)
F N'N
9-2 N \ N 577.2 578.3 2.87
N
NH
N N'S
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EXAMPLE 28
/ o
F\ I N' I 0 / I 0
F N~ F \ N
F H N` I N F H N-N F H N-N
Part A I Part B I
';'~"N \ N
I N\ y Y
NY)--- N N\- N N~ N
N'SEM NsSEM NH
~ S
HO N MsO N-S /-IN N'S
Part A: To a solution of alcohol from Example 26, Part B (30 mg, 0.047 mmol)
in 1.5
mL of THF, was added triethylamine (9.5 mg, 0.094 mmol) followed by
methanesulfonylchloride (7.3 L, 0.094 mmol). The reaction was stirred at room
temperature for 20 min. It was diluted with 10 mL of CH2CI2, washed with 5 mL
of 1 N
aqueous HCI. The organic was dried over anhydrous Na2SO4. The solvent was
removed to give 31 mg of the title compound as a crude material which was used
in
the subsequent reaction without further purification.
Part B: A mixture of mesylate from Part A (9.6 mg, 0.014 mmol), N,N-
diethylisopropylamine (6.0 mg, 0.068 mmol) and Nal (4.1 mg, 0.027 mmol) in 1
mL of
THE was stirred at 60 C for 3 h. It was diluted with 10 mL of CH2CI2 and
washed with
water. The organic was concentrated under vacuum. The residue was purified by
flash chromatography eluting with NH4OH (aq.)/MeOH/CH2CI2 (1:10:190) to give 8
mg
of N-(2,3-difluoro-phenyl)-2-(4-{8-[{3-[(ethyl-isopropyl-amino)-methyl]-
isothiazol-5-yl}-
(2-trimethylsilanyl-ethoxymethyl)-amino)-6-methyl-imidazo[1,2-a]pyrazin-3-yl)-
pyrazol-
1-yl)-acetamide. To a solution of N-(2,3-difluoro-phenyl)-2-(4-{8-[{3-[(ethyl-
isopropyl-
amino)-methyl]-isothiazol-5-yl}-(2-trimethylsilanyl-ethoxymethyl)-amino]-6-
methyl-
imidazo[1,2-a]pyrazin-3-yl}-pyrazol-1-yl)-acetamide (8.0 mg, 0.011 mmol) in 2
mL of
THF heated at 80 C was added 0.2 mL of 4 N HCl in dioxane. The reaction was
stirred at 80 C for 1.5 h. It was cooled to room temperature and then added 8
mL of
ether. The solid was collected by filtration and washed with ether to give 5.8
mg of the
title compound. 1H NMR (400 MHz, CD30D) 6 8.35 (s, 1 H), 7.95-8.10 (m, 3H),
7.70-
7.82 (m, 1 H), 7.25 (s, 1 H), 7.00-7.20 (m, 2H), 5.30 (s, 2H), 4.30-4.60 (m,
2H), 3.75-
3.85 (m, 1 H), 2.60 (s, 3H), 1.30-1.50 (m, 9H). HPLC-MS tR = 2.80 min (UV
254nm).
Mass calculated for formula C27H29F2N9OS 565:2; observed MH+ (LCMS) 566.3
(m/z).
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EXAMPLE 29
N'l o
N~ Part A N' l JO Part B H
l NH2 N" - cl F NN-N
F H
F
11-~
Part A: To a solution of 4-amino-3-fluoro pyridine (560 mg, 5.0 mmol) and Et3N
(760
mg, 7.5 mmol) in 20 mL of THF, was added chloroacetyl chloride (622 mg, 5.5
mmol).
The reaction was stirred at room temperature and monitored by thin layer
chromatography. More chloroacetyl chloride was added until 4-amino-3-fluoro
pyridine was consumed. It was quenched by adding 20 mL of saturated aqueous
NaHCO3. The mixture was diluted with 150 mL of CH2CI2. The organic layer was
concentrated and purified by flash chromatography eluting with 35%
EtOAc/CH2CI2 to
give 850 mg of the title compound. NMR (400 MHz, CDCI3) d 8.62 (brs, 1H), 8.41
(d,
1 H), 8.33 (d, 1 H), 8.26 (t, 1 H), 4.20 (s, 2H).
Part B: A mixture of amide from Part A (106 mg, 0.55 mmol) and Cs2CO3 (326 mg,
1.0
mmol) in 2 mL of DMSO was heated at 100 C for 5 min. To the mixture was added
4-
pyrazoleboronic acid pinacol ester (94 mg, 0.50 mmol). The reaction was
stirred at
100 C for 20 min. It was cooled to room temperature and diluted with 30 mL of
CH2CI2. The mixture was washed with water. The organic was concentrated and
purified by running a quick column eluting with 2% MeOH/EtOAc to give 52 mg of
the
title compound. NMR (400 MHz, CDCI3) d 9.30 (brs, 1H), 8.33 (d, 1H), 8.20-8.30
(m,
2H), 7.93 (s, 1 H), 7.75 (s, 1 H), 4.90 (s, 2H), 1.24 (s, 12H).
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EXAMPLE 30
N`
N
H `
N Pd(PPh3)4 F N
Part B
N.SEM K3PO4, DMF i= N \ -
Part A ~ ~~
HO NS NY~1'N
\ N,SEM
HO N-S
N' O
N~ I o N-'---~
F H N`N
F IN
Part C
N
~N \ N
N
N
NNH
N'SEM N N S
MsO N-S
Part A: To a mixture of iodide (43 mg, 0.083 mmol), boronate from Example 29,
Part B
(43 mg, 0.124 mmol) and Pd(PPh3) 4 (14 mg, 0.012 mmol) in a vial, was added
1.1 mL
of DMF, followed by adding 0.11 mL of 3 M aqueous K3P04 solution (0.33 mmol).
The
vial was sealed and stirred at 65 C overnight. It was diluted with 30 mL of
EtOAc and
washed with water. It was concentrated and purified by flash chromatography
eluting
with 7% MeOH/DCM to give 24 mg of the title compound. NMR (400 MHz, CDCI3) d
9.25 (brs, 1 H), 8.25-8.43 (m, 3H), 7.95 (s, 1 H), 7.80 (s, 1 H), 7.61 (s, 1
H), 7.50 (s, 1.H),
7.09 (s, 1 H), 6.60 (s, 2H), 5.05 (s, 2H), 4.72 (s, 2H), 3.70 (t, 2H), 2.75
(brs, 1 H), 2.48
(s, 3H), 0.90 (t, 2H), -0.14 (s, 9H).
Part B: To a solution of alcohol from Part A (102 mg, 0.17 mmol) in 5 mL of
THF, was
added NEt3 (0.094 mL, 0.67 mmol), followed by methanesulfonylchloride (0.029
mL,
0.37 mmol). The reaction was stirred at room temperature for 15 min. It was
monitored by thin layer chromatography and found starting alcohol was not
totally
consumed. Additional methanesulfonylchloride (0.0035 mL, 0.039 mmol) was
added.
The stirring was continued for 5 min. It was quenched by adding 2 mL of
saturated
NH4CI (aq.) and 2 mL of water. The organic layer was collected. The aqueous
layer
was extracted with CH2CI2 (10 mL X 3) until no desired product remains in
aqueous
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layer. The combined organics were further purified by flash chromatography
eluting
with MeOH/CH2CI2 (1:10) to give 63.3 mg of the title compound as a light
yellow solid.
1H NMR (400 MHz, DMSO-d6) 310.75 (s, 1H), 8.60 (d, 1H), 8.51 (s, 1H), 8.36 (d,
1H),
8.20 (t, 1 H), 8.10 (s, 2H), 7.96 (s, 1 H), 7.38 (s, 1 H), 6.70 (brs, 2H),
5.34 (s, 2H), 5..28
(s, 2H), 3.68 (t, 2H), 3.28 (s, 3H), 2.54 (s, 3H), 0.85 (t, 2H), -0.10 (s,
9H).
Part C: A mixture of mesylate from Part B (24.7 mg, 0.036 mmol), N,N-
diethylisopropylamine (7.8 mg, 0.089 mmol) and Nal (1 mg, 0.007 mmol) in 1.5
mL of
THE was stirred at 80 C for 4 h. It was diluted with 10 mL of CH2CI2 and
washed with
water and brine. It was dried over anhydrous Na2SO4. The organic was
concentrated
under vacuum. The residue was purified by flash chromatography eluting with 7N
NH3 in MeOH /CH2CI2 (1:30) to give 11.5 mg of 2-(4-{8-[{3-[(ethyl-isopropyl-
amino)-
methyl]-isothiazol-5-yl)-(2-trimethylsilanyl-ethoxymethyl)-amino]-6-methyl-
imidazo[1,2-
a]pyrazin-3-yl}-pyrazol-1-yl)-N-(3-fluoro-pyridin-4-yl)-acetamide. NMR (400
MHz,
CDCI3) 6 9.36 (brs, 1 H), 8.30-8.48 (m, 3H), 8.02 (s, 1 H), 7.85 (s, 1 H),
7.66 (s, 1 H),
7.55 (s, 1 H), 7.30 (s, 1 H), 6.62 (s, 2H), 3.60-3.85 (m, 4H), 2.96-3.15 (brs,
3H), 2.40-
2.68 (m, 5H), 0.90-1.20 (m, 12H), 0.00 (s, 9H). To a solution of 2-(4-{8-[{3-
[(ethyl-
isopropyl-amino)-methyl]-isothiazol-5-yi}-(2-trimethylsila nyl-ethoxymethyl )-
amino]-6-
methyl-imidazo[1,2-a]pyrazin-3-yl)-pyrazol-1-yl)-N-(3-fluoro-pyridin-4-yl)-
acetamide
(11.5 mg, 0.0169 mmol) in 0.4 mL of THE heated at 80 C was added 0.4 mL of 4
N
HCI in dioxane. The reaction was stirred at 80 C for 1 h. It was cooled to
room
temperature and then added 8 mL of ether. The solid was collected by
filtration and
washed with ether to give 10 mg of the title compound. HPLC-MS tR = 2.18 min
(UV
254 m). Mass calculated for formula C26H29FNioOS 548.2; observed MH+ (LCMS)
549.3
(m/z).
EXAMPLE 31
By essentially the same procedure set forth in Example 30, only replacing
ethylisopropylamine with other respective amines in Part C, compounds shown in
column 2 of Table 10 were prepared.
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TABLE 10
LCMS
Example Column 2 MW MH' HPLC
m/z MStR
N' I O
NHIE')
F N`N
\ I
N
17-1 N 578.7 579.3 2.17
NH
/-N N-S
OH
N~ O
N Hlfl--A
F N-N
I
17-2 N 578.7 579.3 2.28
N Y `N
NH
/-N N-S
N' 0
NH'--~
F N`N
17-3 N \ 592.7 593.3 2.23
N
NH
N N-S
O-\
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1, O
NHIL-~
F N,N
17-4 N 578.7 579.3 2.28
N Il-- N
NH
/N~N-S
1 OH
N' I O
\ N HL-\
F N'N
17-5 N 604.7 605.3 2.40
N
NH
N N-S
~OH
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ASSAYS:
Aurora Enzyme Assay
An in vitro assay was developed that utilizes recombinant Aurora A or Aurora B
as an enzyme source and a peptide based on PKA as the substrate.
Aurora A Assay:
Aurora A kinase assays were performed in low protein binding 384-well plates
(Corning Inc). All reagents were thawed on ice. Compounds were diluted in 100%
DMSO to desirable concentrations. Each reaction consisted of 8 nM enzyme
(Aurora
A, Upstate cat#14-511), 100 nM Tamra-PKAtide (Molecular Devices, 5TAMRA-
GRTGRRNSICOOH ), 25 pM ATP (Roche), 1 mM DTT (Pierce), and kinase buffer (10
mM Tris, 10 mM MgCI2, 0.01 % Tween 20). For each reaction, 14 l containing
TAMRA-PKAtide, ATP, DTT and kinase buffer were combined with 1 I diluted
compound. The kinase reaction was started by the addition of 5 l diluted
enzyme.
The reaction was allowed to run for 2 hours at room temperature. The reaction
was
stopped by adding 60 l IMAP beads (1:400 beads in progressive (94.7% buffer
A:
5.3% buffer B) 1X buffer, 24 mM NaCI). After an additional 2 hours,
fluorescent
polarization was measured using an Analyst AD (Molecular devices).
Aurora B Assay:
Aurora B kinase assays were performed in low protein binding 384-well plates
(Corning Inc). All reagents were thawed on ice. Compounds were diluted in 100%
DMSO to desirable concentrations. Each reaction consisted of 26 nM enzyme
(Aurora B, Invitrogen cat#pv3970), 100 nM Tamra-PKAtide (Molecular Devices,
5TAMRA-GRTGRRNSICOOH ), 50 M ATP (Roche), 1 mM DTT (Pierce), and kinase
buffer (10 mM Tris, 10 mM MgCl2, 0.01% Tween 20). For each reaction, 14 l
containing TAMRA-PKAtide, ATP, DTT and kinase buffer were combined with 1 l
diluted compound. The kinase reaction was started by the addition of 5 l
diluted
enzyme. The reaction was allowed to run for 2 hours at room temperature. The
reaction was stopped by adding 60 l IMAP beads (1:400 beads in progressive
(94.7% buffer A: 5.3% buffer B) 1 X buffer, 24 mM NaCI). After an additional 2
hours,
fluorescent polarization was measured using an Analyst AD (Molecular devices).
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IC0 Determinations:
Dose-response curves were plotted from inhibition data generated each in
duplicate, from 8 point serial dilutions of inhibitory compounds.
Concentration of
compound was plotted against kinase activity, calculated by degree of
fluorescent
polarization. To generate IC50 values, the dose-response curves were then
fitted to a
standard sigmoidal curve and IC50 values were derived by nonlinear regression
analysis.
Several compounds of the present invention exhibit Aurora A IC50 values of
about 0.0001 nm to about 4 nm, Aurora B IC50 values of about 0.0001 nm to
about 13
nM, and p-HH3 IC50 values of about 1 nM to about 10,000 nM. Additional
compounds
exhibit Aurora A IC50 values of about 0.0001 nm to about 3000 nm, Aurora B
IC50
values of about 0.0001 nm to about 3000 nM, and p-HH3 IC50 values of about 1
nM to
about 10,000 nM.
While the present invention has been described in conjunction with the
specific
embodiments set forth above, many alternatives, modifications and other
variations
thereof will be apparent. to those of ordinary skill in the art. All such
alternatives,
modifications and variations are intended to fall within the spirit and scope
of the
present invention.
