U.S. patent application number 13/452100 was filed with the patent office on 2012-08-16 for pyrrolopyrimidine compounds and their uses.
This patent application is currently assigned to ASTEX THERAPEUTICS LIMITED. Invention is credited to Christopher Thomas BRAIN, Moo Je SUNG, Gebhard THOMA.
Application Number | 20120207763 13/452100 |
Document ID | / |
Family ID | 38779335 |
Filed Date | 2012-08-16 |
United States Patent
Application |
20120207763 |
Kind Code |
A1 |
BRAIN; Christopher Thomas ;
et al. |
August 16, 2012 |
PYRROLOPYRIMIDINE COMPOUNDS AND THEIR USES
Abstract
The present application describes organic compounds that are
useful for the treatment, prevention and/or amelioration of
diseases, particularly pyrrolopyrimidine compounds and derivatives
are described which inhibit protein kinases. The organic compounds
are useful in treating proliferative disease.
Inventors: |
BRAIN; Christopher Thomas;
(North Reading, MA) ; SUNG; Moo Je; (Belmont,
MA) ; THOMA; Gebhard; (Lorrach, DE) |
Assignee: |
ASTEX THERAPEUTICS LIMITED
Cambridge
GB
NOVARTIS AG
Basel
CH
|
Family ID: |
38779335 |
Appl. No.: |
13/452100 |
Filed: |
April 20, 2012 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12302223 |
Aug 24, 2009 |
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PCT/US2007/069595 |
May 24, 2007 |
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13452100 |
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60808605 |
May 26, 2006 |
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Current U.S.
Class: |
424/144.1 ;
424/130.1; 424/158.1; 424/649; 514/171; 514/19.3; 514/19.4;
514/19.5; 514/19.9; 514/234.2; 514/249; 514/252.16; 514/265.1;
514/27; 514/34 |
Current CPC
Class: |
A61P 35/02 20180101;
A61P 35/00 20180101; A61P 9/00 20180101; A61P 3/10 20180101; A61P
25/00 20180101; A61P 17/06 20180101; A61P 17/10 20180101; A61P
25/14 20180101; A61P 19/10 20180101; C07D 487/04 20130101; A61K
31/522 20130101; A61P 3/00 20180101; A61P 11/06 20180101; A61P
19/02 20180101; A61P 29/00 20180101; A61P 37/06 20180101; A61P
43/00 20180101; A61K 31/52 20130101; A61P 15/00 20180101; A61P
37/08 20180101; A61P 37/00 20180101; A61P 5/14 20180101; A61P 15/06
20180101; A61P 25/28 20180101; A61K 31/519 20130101; A61P 1/00
20180101; A61P 1/02 20180101; A61P 17/00 20180101; A61P 1/04
20180101 |
Class at
Publication: |
424/144.1 ;
514/265.1; 514/252.16; 514/234.2; 514/19.9; 424/130.1; 514/19.4;
514/171; 424/158.1; 424/649; 514/34; 514/249; 514/27; 514/19.5;
514/19.3 |
International
Class: |
A61K 31/519 20060101
A61K031/519; A61P 35/00 20060101 A61P035/00; A61K 38/13 20060101
A61K038/13; A61K 38/16 20060101 A61K038/16; A61K 31/56 20060101
A61K031/56; A61K 33/24 20060101 A61K033/24; A61K 31/704 20060101
A61K031/704; A61K 31/7048 20060101 A61K031/7048; A61K 31/5377
20060101 A61K031/5377; A61K 39/395 20060101 A61K039/395 |
Claims
1-43. (canceled)
44. A method for the treatment of cancer comprising administration
of an effective amount of a compound according to formula (I) or a
pharmaceutically acceptable salt thereof to a subject in need of
treatment thereof ##STR00653## wherein: the dashed line indicates a
double bond; A is N; R.sup.2 is hydrogen and R.sup.3 is selected
from the group consisting of hydrogen, hydroxyl,
C.sub.1-C.sub.3-alkyl, C.sub.3-C.sub.8-cycloalkyl, heterocyclyl,
aryl, heteroaryl, substituted C.sub.1-C.sub.3-alkyl, substituted
C.sub.3-C.sub.8-cycloalkyl, substituted heterocyclyl, substituted
aryl and substituted heteroaryl; R.sup.4 is selected from the group
consisting of hydrogen, branched C.sub.1-C.sub.5-alkyl, branched
C.sub.1-C.sub.5-alkyl substituted by phenyl and
C.sub.3-C.sub.6-cycloalkyl; X is CR.sup.11 and Y is CR.sup.12;
R.sup.11 is hydrogen or C.sub.1-C.sub.3-alkyl and R.sup.12 is
BC(O)NR.sup.13R.sup.14; wherein B is a bond, C.sub.1-C.sub.3-alkyl
or branched C.sub.1-C.sub.3-alkyl; wherein R.sup.13 and R.sup.14
are each, independently, selected from the group consisting of
hydrogen, C.sub.1-C.sub.3-alkyl, C.sub.3-C.sub.8-cycloalkyl,
heterocyclyl, aryl, heteroaryl, substituted alkyl, substituted
cycloalkyl, substituted heterocyclyl, substituted aryl, and
substituted heteroaryl.
45. The method of claim 44, wherein R.sup.4 is
C(H)(CH.sub.2CH.sub.3).sub.2, C(H)(CH.sub.2CH.sub.3)Ph,
CH.sub.2CH.sub.3, cyclopropyl, cyclopentyl or cyclohexyl.
46. The method of claim 44, wherein R.sup.3 is an aryl group, which
is further independently substituted one or more times by halogen,
C.sub.1-C.sub.4-alkoxy, R.sup.15-amine, R.sup.15-heterocycle, or
R.sup.15-heteroaryl, wherein R.sup.15 is a bond, C(O), N(H)C(O),
N(H)SO.sub.2, OC(O) or (CH.sub.2).sub.1-4, wherein the
(CH.sub.2).sub.14 group may be interrupted by O, N(CH.sub.3) or
N(H).
47. The method of claim 46, wherein the aryl group is phenyl.
48. The method of claim 44, wherein R.sup.3 is phenyl which is
further independently substituted one or more times with fluoro,
methoxy, diethylamine, R.sup.15-piperazinyl, R.sup.15-morpholinyl,
R.sup.15-piperidinyl, R.sup.15-triazolyl, R.sup.15-phenyl,
R.sup.15-pyridinyl, R.sup.15-piperazinyl, R.sup.15-indazolyl,
R.sup.15-pyrrolidinyl or R.sup.15-imidazolyl, wherein the
piperazinyl, morpholinyl, piperidinyl, triazolyl, phenyl,
pyridinyl, piperazinyl, indazolyl, pyrrolidinyl or imidazolyl
groups may be further substituted with C.sub.1-C.sub.4-alkyl,
C(O)C.sub.1-C.sub.4-alkyl, S(O).sub.2C.sub.1-C.sub.4-alkyl, OH,
C(O)(CH.sub.2).sub.1-3CN or N(H)C(O)C.sub.1-C.sub.4-alkyl and
wherein R.sup.15 is a bond, C(O), N(H)C(O), N(H)SO.sub.2, OC(O) or
(CH.sub.2).sub.14, wherein the (CH.sub.2).sub.14 group may be
interrupted by O, N(CH.sub.3) or N(H).
49. The method of claim 44, wherein R.sup.3 is phenyl which is
further substituted by N(H)C(O)aryl, C(O)N(H)C.sub.1-C.sub.4-alkyl,
C(O)N(C.sub.1-C.sub.4-alkyl).sub.2 or
C(O)N(H)C.sub.3-C.sub.6-cycloalkyl.
50. The method of claim 44, wherein the compound is selected from
the group consisting of ##STR00654## ##STR00655## ##STR00656##
##STR00657## ##STR00658## ##STR00659## ##STR00660## ##STR00661##
##STR00662##
51. The method according to claim 44 wherein the cancer is selected
from the group consisting of breast, stomach, ovary, colon, lung,
brain, larynx, lymphatic system, genitourinary tract, ovarian,
gastric, bone, and pancreatic cancer.
52. The method according to claim 44 wherein the cancer is selected
from the group consisting of bladder, head and neck, breast,
stomach, ovary, colon, lung, brain, larynx, lymphatic system,
genitourinary tract, gastrointestinal, ovarian, prostate, gastric,
bone, small-cell lung, glioma, colorectal and pancreatic
cancer.
53. The method according to claim 44 wherein the compound or
pharmaceutically acceptable salt thereof is administered
simultaneously or sequentially with an anti-inflammatory,
antiproliferative, chemotherapeutic, immunosuppressant,
anti-cancer, or cytotoxic agent.
54. The method according to claim 44 wherein the compound or
pharmaceutically acceptable salt thereof is administered
simultaneously or sequentially with a PTK inhibitor, cyclosporin A,
CTLA4-Ig, an antibody selected from the group consisting of
anti-ICAM-3, anti-IL-2 receptor, anti-CD45RB, anti-CD2, anti-CD3,
anti-CD4, anti-CD80, anti-CD86, and monoclonal antibody OKT3, an
agent blocking the interaction between CD40 and gp39, a fusion
protein constructed from CD40 and gp39, an inhibitor of NF-kappa B
function, a non-steroidal anti-inflammatory drug, a steroid, a gold
compound, an antiproliferative agent, FK506, mycophenolate mofetil,
a cytotoxic drug, a TNF-.alpha. inhibitor, an anti-TNF antibody,
soluble TNF receptor, rapamycin, leflunimide, cyclooxygenase-2
inhibitor, paclitaxel, cisplatin, carboplatin, doxorubicin,
caminomycin, daunorubicin, aminopterin, methotrexate, methopterin,
mitomycin C, ecteinascidin 743, porfiromycin, 5-fluorouracil,
6-mercaptopurine, gemcitabine, cytosine arabinoside,
podophyllotoxin, etoposide, etoposide phosphate, teniposide,
melphalan, vinblastine, vincristine, leurosidine, epothilone,
vindesine, or leurosine.
Description
BACKGROUND
[0001] The search for new therapeutic agents has been greatly aided
in recent years by a better understanding of the structure of
enzymes and other biomolecules associated with diseases. One
important class of enzymes that has been the subject of extensive
study is protein kinases.
[0002] Protein kinases constitute a large family of structurally
related enzymes that are responsible for the control of a variety
of signal transduction processes within the cell. (Hardie, G. and
Hanks, S. The Protein Kinase Facts Book, I and II, Academic Press,
San Diego, Calif.: 1995). Protein kinases are thought to have
evolved from a common ancestral gene due to the conservation of
their structure and catalytic function. Almost all kinases contain
a similar 250-300 amino acid catalytic domain. The kinases may be
categorized into families by the substrates they phosphorylate
(e.g., protein-tyrosine, protein-serine/threonine, lipids, etc.).
Sequence motifs have been identified that generally correspond to
each of these kinase families (See, for example, Hanks, S. K.,
Hunter, T., FASEB J. 1995, 9, 576-596; Knighton et al., Science
1991, 253, 407-414; Hiles et al., Cell 1992, 70, 419-429; Kunz et
al., Cell 1993, 73, 585-596; Garcia-Bustos et al., EMBO J. 1994,
13, 2352-2361).
[0003] In general, protein kinases mediate intracellular signaling
by affecting a phosphoryl transfer from a nucleoside triphosphate
to a protein acceptor that is involved in a signaling pathway.
These phosphorylation events act as molecular on/off switches that
can modulate or regulate the target protein biological function.
These phosphorylation events are ultimately triggered in response
to a variety of extracellular and other stimuli. Examples of such
stimuli include environmental and chemical stress signals (e.g.,
osmotic shock, heat shock, ultraviolet radiation, bacterial
endotoxin, and H.sub.2O.sub.2), cytokines (e.g., interleukin-1
(IL-1) and tumor necrosis factor-.alpha. (TNF-.alpha.)), and growth
factors (e.g., granulocyte macrophage-colony-stimulating factor
(GM-CSF), and fibroblast growth factor (FGF)). An extracellular
stimulus may affect one or more cellular responses related to cell
growth, migration, differentiation, secretion of hormones,
activation of transcription factors, muscle contraction, glucose
metabolism, control of protein synthesis, and regulation of the
cell cycle.
[0004] Many diseases are associated with abnormal cellular
responses triggered by protein kinase-mediated events as described
above. These diseases include, but are not limited to, autoimmune
diseases, inflammatory diseases, bone diseases, metabolic diseases,
neurological and neurodegenerative diseases, cancer, cardiovascular
diseases, allergies and asthma, Alzheimer's disease, and
hormone-related diseases. Accordingly, there has been a substantial
effort in medicinal chemistry to find protein kinase inhibitors
that are effective as therapeutic agents.
[0005] The Janus kinases (JAK) are a family of tyrosine kinases
consisting of JAK1, JAK2, JAK3 and TYK2. The JAKs play a critical
role in cytokine signaling. The down-stream substrates of the JAK
family of kinases include the signal transducer and activator of
transcription (STAT) proteins. JAK/STAT signaling has been
implicated in the mediation of many abnormal immune responses such
as allergies, asthma, autoimmune diseases such as transplant
rejection, rheumatoid arthritis, amyotrophic lateral sclerosis and
multiple sclerosis as well as in solid and hematologic malignancies
such as leukemias and lymphomas. The pharmaceutical intervention in
the JAK/STAT pathway has been reviewed [Frank Mol. Med. 5: 432-456
(1999) & Seidel, et al, Oncogene 19: 2645-2656 (2000)].
[0006] JAK1, JAK2, and TYK2 are ubiquitously expressed, while JAK3
is predominantly expressed in hematopoietic cells. JAK3 binds
exclusively to the common cytokine receptor gamma chain
(.gamma..sub.c) and is activated by IL-2, IL-4, IL-7, IL-9, and
IL-15. The proliferation and survival of murine mast cells induced
by IL-4 and IL-9 have, in fact, been shown to be dependent on JAK3-
and 65.sub.c-signaling [Suzuki et al, Blood 96: 2172-2180
(2000)].
[0007] Cross-linking of the high-affinity immunoglobulin (Ig) E
receptors of sensitized mast cells leads to a release of
proinflammatory mediators, including a number of vasoactive
cytokines resulting in acute allergic, or immediate (type I)
hypersensitivity reactions [Gordon et al, Nature 346: 274-276
(1990) & Galli, N. Engl. J. Med., 328: 257-265 (1993)]. A
crucial role for JAK3 in IgE receptor-mediated mast cell responses
in vitro and in vivo has been established [Malaviya, et al,
Biochem. Biophys. Res. Commun. 257: 807-813 (1999)]. In addition,
the prevention of type I hypersensitivity reactions, including
anaphylaxis, mediated by mast cell-activation through inhibition of
JAK3 has also been reported [Malaviya et al, J. Biol. Chem.
274:27028-27038 (1999)].
[0008] The JAK family of tyrosine kinases have also been shown to
play a role in immunosuppression and allograft acceptance [Kirken,
Transpl. Proc. 33: 3268-3270 (2001)], rheumatoid arthritis
[Muller-Ladner, et al., J. Immunol. 164: 3894-3901 (2000)],
Familial amyotrophic lateral sclerosis [Trieu, et al., Biochem.
Biophys. Res. Commun. 267: 22-25 (2000)], and leukemia [Sudbeck, et
al., Clin. Cancer Res. 5: 1569-1582 (1999)].
[0009] Initiation, progression, and completion of the mammalian
cell cycle are regulated by various cyclin-dependent kinase (CDK)
complexes, which are critical for cell growth. These complexes
comprise at least a catalytic (the CDK itself) and a regulatory
(cyclin) subunit. Some of the more important complexes for cell
cycle regulation include cyclin A (CDK1--also known as cdc2, and
CDK2), cyclin B1-B3 (CDK1) and cyclin D1-D3 (CDK2, CDK4, CDK5,
CDK6), cyclin E (CDK2). Each of these complexes is involved in a
particular phase of the cell cycle. Not all members of the CDK
family are involved exclusively in cell cycle control, however.
Thus CDKs 7, 8, and 9 are implicated in the regulation of
transcription, and CDK5 plays a role in neuronal and secretory cell
function.
[0010] The activity of CDKs is regulated post-translationally, by
transitory associations with other proteins, and by alterations of
their intracellular localization. Tumor development is closely
associated with genetic alteration and deregulation of CDKs and
their regulators, suggesting that inhibitors of CDKs may be useful
anti-cancer therapeutics. Indeed, early results suggest that
transformed and normal cells differ in their requirement for, e.g.,
cyclin A/CDK2 and that it may be possible to develop novel
antineoplastic agents devoid of the general host toxicity observed
with conventional cytotoxic and cytostatic drugs. While inhibition
of cell cycle-related CDKs is clearly relevant in, e.g., oncology
applications, this may not be the case for the inhibition of RNA
polymerase-regulating CDKs. On the other hand, inhibition of
CDK9/cyclin T function was recently linked to prevention of HIV
replication and the discovery of new CDK biology thus continues to
open up new therapeutic indications for CDK inhibitors (Sausville,
E. A. Trends Molec. Med. 2002, 8, S32-S37).
[0011] The function of CDKs is to phosphorylate and thus activate
or deactivate certain proteins, including e.g. retinoblastoma
proteins, lamins, histone H1, and components of the mitotic
spindle. The catalytic step mediated by CDKs involves a
phospho-transfer reaction from ATP to the macromolecular enzyme
substrate. Several groups of compounds (reviewed in e.g. Fischer,
P. M. Curr. Opin. Drug Discovery Dev. 2001, 4, 623-634) have been
found to possess anti-proliferative properties by virtue of
CDK-specific ATP antagonism.
[0012] Thus, there is a continued need to find new therapeutic
agents to treat human diseases. Accordingly, there is a great need
to develop inhibitors of protein kinases, such as Jak1, Jak2 and
Jak3, as well as CDK1, CDK2, CDK4, CDK5, CDK6, CDK7, CDK8 and
CDK9.
SUMMARY OF THE INVENTION
[0013] There remains a need for new treatments and therapies for
protein kinase-associated disorders. There is also a need for
compounds useful in the treatment or prevention or amelioration of
one or more symptoms of cancer, transplant rejections, and
autoimmune diseases. Furthermore, there is a need for methods for
modulating the activity of protein kinases, such as Jak1, Jak2 and
Jak3, as well as CDK1, CDK2, CDK4, CDK5, CDK6, CDK7, CDK8 and CDK9,
using the compounds provided herein. In one aspect, the invention
provides a compound of Formula I:
##STR00001##
[0014] In one aspect of the invention, the protein kinase is a
protein tyrosine kinase. In one embodiment, the protein kinase is
selected from the group consisting of abl, ATK, ber-abl, Blk, Brk,
Btk, c-fms, e-kit, c-met, c-src, CDK, cRaf1, CSFIR, CSK, EGFR,
ErbB2, ErbB3, ErbB4, ERK, Fak, fes, FGFRI, 25 FGFR2, FGFR3, FGFR4,
FGFR5, Fgr, FLK-4, flt-1, Fps, Frk, Fyn, GSK, Gst-Flkl, Hck, Her-2,
Her-4, IGF-1R, INS-R, Jak, JNK, KDR, Lck, Lyn, MEK, p38, PANHER,
PDGFR, PLK, PKC, PYK2, Raf, Rho, ros, SRC, t' eII t' e2, TRK, TYK2,
UL97, VEGFR, Yes, and Zap70. In another embodiment, the protein
kinase is selected from the group consisting of CDK1, CDK2, CDK4,
CDK5, CDK6, CDK7, CDK8 and CDK9. In yet another embodiment, the
protein kinase is selected from the group consisting of Jak1, Jak2
and Jak3. In still another embodiment, the protein kinase is
selected from the group consisting of Jak3 and CDK4.
[0015] In another aspect of the invention, the protein kinase is in
a cell culture. In still another aspect, the protein kinase is in a
mammal.
[0016] In another aspect, the invention provides a method of
treating a protein kinase-associated disorder, wherein the method
includes administering to a subject in need thereof a
pharmaceutically acceptable amount of a compound of the Formula I,
such that the protein kinase-associated disorder is treated. In one
embodiment, the protein kinase is selected from the group
consisting of CDK1, CDK2, CDK4, CDK5, CDK6, CDK7, CDK8, CDK9, Jak1,
Jak2 and Jak3. In a particular embodiment, the protein kinase is
selected from the group consisting of Jak3 and CDK4.
[0017] In another embodiment, the protein kinase-associated
disorder is selected from the group consisting of blood vessel
proliferative disorders, fibrotic disorders, mesangial cell
proliferative disorders, metabolic disorders, allergies, asthma,
thrombosis, nervous system diseases and cancer.
[0018] In another embodiment, the protein kinase-associated
disorder is cancer. In yet another embodiment, the cancer is
selected from the group consisting of breast, stomach, ovary,
colon, lung, brain, larynx, lymphatic system, genitourinary tract
(including bladder and prostate), ovarian, gastric, bone, and
pancreatic cancer.
[0019] In another embodiment, the protein kinase-associated
disorder is selected from the group consisting of organ transplant
rejection, xeno transplantation, lupus, multiple sclerosis,
rheumatoid arthritis, psoriasis, Type 1 diabetes and complications
from diabetes, cancer, asthma, atopic dermatitis, autoimmune
thyroid disorders, ulcerative colitis, Crohn's disease, Alzheimer's
disease and leukemia.
[0020] In still another embodiment, the disease is selected from an
immune response, an autoimmune disease, a neurodegenerative
disease, or a solid or hematologic malignancy. In yet another
embodiment, the disease is selected from an allergic or type I
hypersensitivity reaction, asthma, graft versus host disease,
rheumatoid arthritis, amyotrophic lateral sclerosis, multiple
sclerosis, Familial amyotrophic lateral sclerosis, leukemia, or
lymphoma
[0021] In another aspect, the invention provides a method of
treating an autoimmune disease, wherein the treatment includes
administering to a subject in need thereof a pharmaceutically
acceptable amount of a compound of the Formula I, such that the
autoimmune disease is treated. In one embodiment, the autoimmune
disease is selected from the group consisting of autoimmune
hemolytic anemia, autoimmune neonatal thrombocytopenia, idiopathic
thrombocytopenia purpura, autoimmunocytopenia, hemolytic anemia,
antiphospholipid syndrome, dermatitis, allergic encephalomyelitis,
myocarditis, relapsing polychondritis, rheumatic heart disease,
glomerulonephritis, multiple sclerosis, neuritis, uveitis
ophthalmia, polyendocrinopathies, purpura, Reiter's Disease,
Stiff-Man Syndrome, autoimmune pulmonary inflammation, autism,
Guillain-Barre Syndrome, insulin dependent diabetes mellitis,
autoimmune inflammatory eye, autoimmune thyroiditis,
hypothyroidism, systemic lupus erhythematosus, Goodpasture's
syndrome, Pemphigus, Receptor autoimmunities, autoimmune hemolytic
anemia, autoimmune thrombocytopenic purpura, rheumatoid arthritis,
mixed connective tissue disease, polymyositis/dermatomyositis,
pernicious anemia, idiopathic Addison's disease, infertility,
glomerulonephritis, bullous pemphigoid, Sjogren's syndrome,
diabetes millitus, adrenergic drug resistance, chronic active
hepatitis, primary biliary cirrhosis, vitiligo, vasculitis,
post-MI, cardiotomy syndrome, urticaria, atopic dermatitis, asthma,
inflammatory myopathies, chronic active hepatitis, primary biliary
cirrhosis and T-cell mediated hypersensitivity diseases.
[0022] In another aspect, the invention provides a method of
treating transplant rejection, wherein the treatment includes
administering to a subject in need thereof a pharmaceutically
acceptable amount of a compound of the Formula I such that the
transplant rejection is treated. In one embodiment, the transplant
rejection is selected from the group consisting of graft versus
host disease, rejection related to xeno transplantation, rejection
related to organ transplant, rejection related to acute transplant,
heterograft or homograft rejection and ischemic or reperfusion
injury incurred during organ transplantation.
[0023] In another aspect, the invention provides a method of
treating cancer, wherein the method includes administering to a
subject in need thereof a pharmaceutically acceptable amount of a
compound of the Formula I such that the cancer disease or disorder
is treated. In one embodiment, the cancer is selected from the
group consisting of bladder, head and neck, breast, stomach, ovary,
colon, lung, brain, larynx, lymphatic system, genitourinary tract,
gastrointestinal, ovarian, prostate, gastric, bone, small-cell
lung, glioma, colorectal and pancreatic cancer.
[0024] In another aspect of the invention, the Formula I or salt
thereof is administered, simultaneously or sequentially, with an
antiinflammatory, antiproliferative, chemotherapeutic agent,
immunosuppressant, anti-cancer, cytotoxic agent or kinase inhibitor
other than a compound of the Formula I or salt thereof. In one
embodiment, the compound of the Formula I or salt thereof is
administered, simultaneously or sequentially, with one or more of a
PTK inhibitor, cyclosporin A, CTLA4-Ig, antibodies selected from
anti-ICAM-3, anti-IL-2 receptor, anti-CD45RB, anti-CD2, anti-CD3,
anti-CD4, anti-CD80, anti-CD86, and monoclonal antibody OKT3,
agents blocking the interaction between CD40 and gp39, fusion
proteins constructed from CD40 and gp39, inhibitors of NF-kappa B
function, non-steroidal antiinflammatory drugs, steroids, gold
compounds, antiproliferative agents, FK506, mycophenolate mofetil,
cytotoxic drugs, TNF-.alpha. inhibitors, anti-TNF antibodies or
soluble TNF receptor, rapamycin, leflunimide, cyclooxygenase-2
inhibitors, paclitaxel, cisplatin, carboplatin, doxorubicin,
caminomycin, daunorubicin, aminopterin, methotrexate, methopterin,
mitomycin C, ecteinascidin 743, porfiromycin, 5-fluorouracil,
6-mercaptopurine, gemcitabine, cytosine arabinoside,
podophyllotoxin, etoposide, etoposide phosphate, teniposide,
melphalan, vinblastine, vincristine, leurosidine, epothilone,
vindesine, leurosine, or derivatives thereof.
[0025] In another aspect, the invention provides a packaged protein
kinase-associated disorder treatment, wherein the treatment
includes a protein kinase-modulating compound of the Formula I,
packaged with instructions for using an effective amount of the
protein kinase-modulating compound to treat a protein
kinase-associated disorder.
DETAILED DESCRIPTION OF THE INVENTION
[0026] This invention is directed to compounds, e.g.,
pyrrolopyrimidine compounds, and intermediates thereto, as well as
pharmaceutical compositions containing the compounds for use in
treatment of protein kinase-associated disorders. This invention is
also directed to the compounds of the invention or compositions
thereof as modulators of Jak 1, Jak2 and Jak3, as well as CDK1,
CDK2, CDK4, CDK5, CDK6, CDK7, CDK8 and CDK9. The present invention
is also directed to methods of combination therapy for inhibiting
protein kinase activity in cells, or for treating, preventing or
ameliorating of one or more symptoms of cancer, transplant
rejections, and autoimmune diseases in patients using the compounds
of the invention or pharmaceutical compositions, or kits
thereof.
[0027] In one aspect, the invention provides compounds of the
Formula I:
##STR00002##
or a pharmaceutically acceptable salt or solvate thereof,
wherein:
[0028] the dashed line indicates a single or double bond;
[0029] A is N or CR.sup.5, wherein R.sup.5 is hydrogen or
C.sub.1-C.sub.3-alkyl;
[0030] R.sup.2 and R.sup.3 are each, independently, selected from
the group consisting of hydrogen, hydroxyl, C.sub.1-C.sub.3-alkyl,
C.sub.3-C.sub.8-cycloalkyl, heterocyclyl, aryl, heteroaryl,
substituted C.sub.1-C.sub.3-alkyl, substituted
C.sub.3-C.sub.8-cycloalkyl, substituted heterocyclyl, substituted
aryl and substituted heteroaryl;
[0031] R.sup.4 is selected from the group consisting of hydrogen,
C.sub.1-C.sub.8-alkyl, substituted C.sub.1-C.sub.8-alkyl,
C.sub.3-C.sub.8-cycloalkyl, substituted C.sub.3-C.sub.8-cycloalkyl,
aryl, substituted aryl, heteroaryl and substituted heteroaryl;
[0032] when the bond between X and Y is a single bond, X is
CR.sup.6R.sup.7, NR.sup.8 or C.dbd.O, and Y is CR.sup.9R.sup.10 or
C.dbd.O;
[0033] when the bond between X and Y is a double bond, X is N or
CR.sup.11, and Y is CR.sup.12;
[0034] wherein R.sup.6 and R.sup.7 are each, independently selected
from the group consisting of aryl, substituted aryl, heteroaryl,
substituted heteroaryl, hydrogen, C.sub.1-C.sub.3-alkyl,
C.sub.3-C.sub.8-cycloalkyl, heterocyclyl, substituted alkyl,
substituted cycloalkyl, and substituted heterocyclyl;
[0035] R.sup.8 is hydrogen, C.sub.1-C.sub.3-alkyl, and
C.sub.3-C.sub.8-cycloalkyl;
[0036] R.sup.9 and R.sup.10 are each, independently, hydrogen,
C.sub.1-C.sub.3-alkyl, or C.sub.3-C.sub.8-cycloalkyl;
[0037] R.sup.11 and R.sup.12 are each, independently, selected from
the group consisting of halo, hydrogen, C.sub.1-C.sub.3-alkyl,
C.sub.1-C.sub.3-alkoxy, CN, C.dbd.NOH, C.dbd.NOCH.sub.3, C(O)H,
C(O)C.sub.1-C.sub.3-alkyl, C.sub.3-C.sub.8-cycloalkyl,
heterocyclyl, aryl, heteroaryl, substituted C.sub.1-C.sub.3-alkyl,
substituted C.sub.3-C.sub.8-cycloalkyl, substituted heterocyclyl,
substituted aryl, substituted heteroaryl, --BNR.sup.13R.sup.14,
--BOR.sup.13, --BC(O)R.sup.13, --BC(O)OR.sup.13,
--BC(O)NR.sup.13R.sup.14; wherein B is a bond,
C.sub.1-C.sub.3-alkyl or branched C.sub.1-C.sub.3-alkyl; wherein
R.sup.13 and R.sup.14 are each, independently, selected from the
group consisting of hydrogen, C.sub.1-C.sub.3-alkyl,
C.sub.3-C.sub.8-cycloalkyl, heterocyclyl, aryl, heteroaryl,
substituted alkyl, substituted cycloalkyl, substituted
heterocyclyl, substituted aryl, and substituted heteroaryl.
[0038] In one embodiment, R.sup.4 is branched or linear
C.sub.1-C.sub.5-alkyl, wherein the branched C.sub.1-C.sub.5-alkyl
group may be interrupted by one or more heteroatoms, and/or
substituted with one or more heteroatoms, halogens, C.sub.3-C.sub.8
cycloalkyl groups, substituted C.sub.3-C.sub.8 cycloalkyl groups,
C.sub.3-C.sub.8 hetrocyclyl groups, aryl groups, heteroaryl groups,
substituted aryl groups, or substituted heteroaryl groups.
[0039] In another embodiment, R.sup.12 is not hydrogen, R.sup.4 is
selected from the group consisting of hydrogen,
C.sub.1-C.sub.8-alkyl, C.sub.3-C.sub.8-cycloalkyl,
C.sub.3-C.sub.8-substituted cycloalkyl, aryl, substituted aryl,
heteroaryl, and substituted heteroaryl.
[0040] In still another embodiment, R.sup.12 is not hydrogen,
R.sup.4 is branched or linear C.sub.1-C.sub.5-alkyl, wherein the
branched C.sub.1-C.sub.5-alkyl group may be interrupted by one or
more heteroatoms, and/or substituted with one or more heteroatoms,
halogens, C.sub.3-C.sub.8 cycloalkyl groups, substituted
C.sub.3-C.sub.8 cycloalkyl groups, C.sub.3-C.sub.8 hetrocyclyl
groups, aryl groups, heteroaryl groups, substituted aryl groups, or
substituted heteroaryl groups.
[0041] In yet another embodiment, A is N.
[0042] In another embodiment, R.sup.4 is selected from the group
consisting of hydrogen, branched C.sub.1-C.sub.5-alkyl, branched
C.sub.1-C.sub.5-alkyl substituted by phenyl and
C.sub.3-C.sub.6-cycloalkyl.
[0043] In yet another embodiment, R.sup.4 is
C(H)(CH.sub.2CH.sub.3).sub.2, C(H)(CH.sub.2CH.sub.3)Ph,
CH.sub.2CH.sub.3, cyclopropyl, cyclopentyl or cyclohexyl.
[0044] In still another embodiment, the dashed line is a single
bond, X is CH.sub.2, C(C.sub.1-C.sub.3-alkyl).sub.2 or
N(C.sub.1-C.sub.3-alkyl), and Y is C.dbd.O. In another embodiment,
X is CH.sub.2 or C(CH.sub.3).sub.2 and Y is C.dbd.O. In yet another
embodiment, the dashed line is a double bond, X is CH, N,
C--C(O)C.sub.1-C.sub.3-alkyl or C--(C.sub.1-C.sub.3-alkyl), and Y
is CH, C--CHO, C--C.sub.1-C.sub.3-alkyl, C--C.sub.1-C.sub.3-alkoxy,
C--C(O)C.sub.1-C.sub.3-alkyl, C--C.dbd.NOH or
C--C.dbd.NOCH.sub.3.
[0045] In another embodiment, R.sup.2 is H.
[0046] In yet another embodiment, R.sup.3 is an aryl group, which
is further independently substituted one or more times by halogen,
C.sub.1-C.sub.4-alkoxy, R.sup.15-amine, R.sup.15-heterocycle, or
R.sup.15-heteroaryl, wherein R.sup.15 is a bond, C(O), N(H)C(O),
N(H)SO.sub.2, OC(O) or (CH.sub.2).sub.1-4, wherein the
(CH.sub.2).sub.1-4 group may be interrupted by O, N(CH.sub.3) or
N(H).
[0047] In still another embodiment, the aryl group is phenyl.
[0048] In another embodiment, the phenyl group is independently
substituted one or more times with fluoro, methoxy, diethylamine,
R.sup.15-piperazinyl, R.sup.15-morpholinyl, R.sup.15-piperidinyl,
R.sup.15-triazolyl, R.sup.15-phenyl, R.sup.15-pyridinyl,
R.sup.15-piperazinyl, R.sup.15-indazolyl, R.sup.15-pyrrolidinyl or
R.sup.15-imidazolyl, wherein the piperazinyl, morpholinyl,
piperidinyl, triazolyl, phenyl, pyridinyl, piperazinyl, indazolyl,
pyrrolidinyl or imidazolyl groups may be further substituted with
C.sub.1-C.sub.4-alkyl, C(O)C.sub.1-C.sub.4-alkyl,
S(O).sub.2C.sub.1-C.sub.4-alkyl, OH, C(O)(CH.sub.2).sub.1-3CN or
N(H)C(O)C.sub.1-C.sub.4-alkyl.
[0049] In yet another embodiment, the phenyl group is substituted
by N(H)C(O)aryl, C(O)N(H)C.sub.1-C.sub.4-alkyl,
C(O)N(C.sub.1-C.sub.4-alkyl).sub.2 or
C(O)N(H)C.sub.3-C.sub.6-cycloalkyl.
[0050] Preferred embodiments of Formula I (including
pharmaceutically acceptable salts thereof, as well as enantiomers,
stereoisomers, rotamers, tautomers, diastereomers, atropisomers or
racemates thereof) are shown below in Table A, Table B, Table C and
Table D, and are also considered to be "compounds of the
invention." The compounds of the invention are also referred to
herein as "protein kinase inhibitors."
TABLE-US-00001 TABLE A Jak-3/ IC50(nM) ##STR00003## * ##STR00004##
* ##STR00005## ** ##STR00006## ** ##STR00007## ** ##STR00008## **
##STR00009## ** ##STR00010## * ##STR00011## ** ##STR00012## **
##STR00013## ** ##STR00014## ** ##STR00015## ** ##STR00016## **
##STR00017## ** ##STR00018## ** ##STR00019## ** ##STR00020## **
##STR00021## ** ##STR00022## ** ##STR00023## ** ##STR00024## **
##STR00025## * ##STR00026## ** ##STR00027## ** ##STR00028## **
##STR00029## ** ##STR00030## ** ##STR00031## ** ##STR00032## **
##STR00033## ** ##STR00034## ** ##STR00035## ** ##STR00036## **
##STR00037## ** ##STR00038## * ##STR00039## * ##STR00040## **
##STR00041## * ##STR00042## * ##STR00043## * ##STR00044## **
##STR00045## * ##STR00046## ** ##STR00047## * ##STR00048## *
##STR00049## ** ##STR00050## ** ##STR00051## * ##STR00052## *
##STR00053## ** ##STR00054## ** ##STR00055## ** ##STR00056## *
##STR00057## ** ##STR00058## ** ##STR00059## ** ##STR00060## **
##STR00061## ** ##STR00062## * ##STR00063## ** ##STR00064## **
##STR00065## * ##STR00066## * ##STR00067## * ##STR00068## **
##STR00069## ** ##STR00070## * ##STR00071## * ##STR00072## **
##STR00073## * ##STR00074## ** ##STR00075## ** ##STR00076## **
##STR00077## ** ##STR00078## ** ##STR00079## ** ##STR00080## *
##STR00081## ** ##STR00082## ** ##STR00083## * ##STR00084## *
##STR00085## ** ##STR00086## ** Table A Key * .ltoreq. 100 nM 100
nM .ltoreq. **
TABLE-US-00002 TABLE B Jak-3 Lance/ IC50 [nmol I-1] ##STR00087##
##STR00088## ##STR00089## ##STR00090## ##STR00091## ##STR00092##
##STR00093## ##STR00094## ##STR00095## ##STR00096## ##STR00097## **
##STR00098## ##STR00099## * ##STR00100## ** ##STR00101## **
##STR00102## ** ##STR00103## ** ##STR00104## ##STR00105## **
##STR00106## ##STR00107## ##STR00108## ##STR00109## ##STR00110##
##STR00111## ** ##STR00112## ** ##STR00113## ##STR00114##
##STR00115## ** ##STR00116## ** ##STR00117## ** ##STR00118## *
##STR00119## * ##STR00120## ** ##STR00121## ** ##STR00122## **
##STR00123## ** ##STR00124## ** ##STR00125## * ##STR00126##
##STR00127## ** ##STR00128## ##STR00129## ##STR00130## ##STR00131##
##STR00132## ##STR00133## ##STR00134## ##STR00135## ##STR00136##
##STR00137## ##STR00138## ##STR00139## ##STR00140## ##STR00141##
##STR00142## ##STR00143## ##STR00144## ##STR00145## ##STR00146##
Table B Key * .ltoreq. 100 nmol.sup.-1 100 nmol.sup.-1 .ltoreq.
**
TABLE-US-00003 TABLE C CDK4 CDK2 IC50, .mu.M IC50, .mu.M
##STR00147## ** * ##STR00148## * ##STR00149## * * ##STR00150## * *
##STR00151## * ** ##STR00152## ** ** ##STR00153## * ** ##STR00154##
* ** ##STR00155## * * ##STR00156## ** ##STR00157## ** ##STR00158##
** ##STR00159## ** ##STR00160## * ##STR00161## ** ##STR00162## *
##STR00163## ##STR00164## ##STR00165## * ##STR00166## *
##STR00167## * ##STR00168## * ##STR00169## * ##STR00170## * Table C
Key * .ltoreq. 10 .mu.M 10 .mu.M .ltoreq. **
TABLE-US-00004 TABLE D ##STR00171## ##STR00172## ##STR00173##
##STR00174## ##STR00175## ##STR00176## ##STR00177## ##STR00178##
##STR00179## ##STR00180##
[0051] In certain embodiments, the compound of the present
invention is further characterized as a modulator of a protein
kinase, including, but not limited to, protein kinases selected
from the group consisting of abl, ATK, ber-abl, Blk, Brk, Btk,
c-fms, e-kit, c-met, c-src, CDK, cRaf1, CSFIR, CSK, EGFR, ErbB2,
ErbB3, ErbB4, ERK, Fak, fes, FGFR1, 25 FGFR2, FGFR3, FGFR4, FGFR5,
Fgr, FLK-4, flt-1, Fps, Frk, Fyn, GSK, Gst-Flkl, Hck, Her-2, Her-4,
IGF-1R, INS-R, Jak, JNK, KDR, Lck, Lyn, MEK, p38, PANHER, PDGFR,
PLK, PKC, PYK2, Raf, Rho, ros, SRC, t' eII t' e2, TRK, TYK2, UL97,
VEGFR, Yes, and Zap70.
[0052] In a preferred embodiment, the protein kinase is selected
from the group consisting of CDK1, CDK2, CDK4, CDK5, CDK6, CDK7,
CDK8 and CDK9. In another preferred embodiment, the protein kinase
is selected from the group consisting of Jak1, Jak2 and Jak3. In a
particularly preferred embodiment, the protein kinase is selected
from the group consisting of Jak3 and CDK4.
[0053] In other embodiments, the compounds of the present invention
are used for the treatment of protein kinase-associated disorders.
As used herein, the term "protein kinase-associated disorder"
includes disorders and states (e.g., a disease state) that are
associated with the activity of a protein kinase, e.g., CDK4 and
Jak3. Non-limiting examples of a protein kinase-associated disorder
include blood vessel proliferative disorders, fibrotic disorders,
mesangial cell proliferative disorders, metabolic disorders,
allergies, asthma, thrombosis, nervous system diseases, organ
transplant rejection, autoimmune diseases, and cancer. In another
embodiment, the compound of the present invention is further
characterized as a modulator of a combination of protein kinases,
e.g., Jak3 and CDK4.
[0054] In certain embodiments, a compound of the present invention
is used for protein kinase-associated diseases, and use of the
compound of the present invention as an inhibitor of any one or
more protein kinases. It is envisioned that a use can be a
treatment of inhibiting one or more isoforms of protein
kinases.
[0055] The compounds of the invention are inhibitors of
cyclin-dependent kinase enzymes (CDKs). Without being bound by
theory, inhibition of the CDK4/cyclin D1 complex blocks
phosphorylation of the Rb/inactive E2F complex, thereby preventing
release of activated E2F and ultimately blocking E2F-dependent DNA
transcription. This has the effect of inducing G.sub.1 cell cycle
arrest. In particular, the CDK4 pathway has been shown to have
tumor-specific deregulation and cytotoxic effects.
[0056] Furthermore, the compounds of this invention have the
potential to block the expansion of auto- or alloreactive T cells,
and thus have beneficial effects on autoimmune diseases, as well as
transplant rejections.
[0057] The present invention includes treatment of one or more
symptoms of cancer, transplant rejections, and autoimmune diseases,
as well as protein kinase-associated disorders, as described above,
but the invention is not intended to be limited to the manner by
which the compound performs its intended function of treatment of a
disease. The present invention includes treatment of diseases
described herein in any manner that allows treatment to occur,
e.g., cancer, transplant rejections, and autoimmune diseases.
[0058] In certain embodiments, the invention provides a
pharmaceutical composition of any of the compounds of the present
invention. In a related embodiment, the invention provides a
pharmaceutical composition of any of the compounds of the present
invention and a pharmaceutically acceptable carrier or excipient of
any of these compounds. In certain embodiments, the invention
includes the compounds as novel chemical entities.
[0059] In one embodiment, the invention includes a packaged protein
kinase-associated disorder treatment. The packaged treatment
includes a compound of the invention packaged with instructions for
using an effective amount of the compound of the invention for an
intended use.
[0060] The compounds of the present invention are suitable as
active agents in pharmaceutical compositions that are efficacious
particularly for treating protein kinase-associated disorders,
e.g., cancer, transplant rejections, and autoimmune diseases. The
pharmaceutical composition in various embodiments has a
pharmaceutically effective amount of the present active agent along
with other pharmaceutically acceptable excipients, carriers,
fillers, diluents and the like. The phrase, "pharmaceutically
effective amount" as used herein indicates an amount necessary to
administer to a host, or to a cell, issue, or organ of a host, to
achieve a therapeutic result, especially the regulating,
modulating, or inhibiting protein kinase activity, e.g., inhibition
of the activity of a protein kinase, or treatment of cancer,
transplant rejections, or autoimmune diseases.
[0061] In other embodiments, the present invention provides a
method for inhibiting the activity of a protein kinase. The method
includes contacting a cell with any of the compounds of the present
invention. In a related embodiment, the method further provides
that the compound is present in an amount effective to selectively
inhibit the activity of a protein kinase.
[0062] In other embodiments, the present invention provides a use
of any of the compounds of the invention for manufacture of a
medicament to treat cancer, transplant rejections, or autoimmune
diseases in a subject.
[0063] In other embodiments, the invention provides a method of
manufacture of a medicament, including formulating any of the
compounds of the present invention for treatment of a subject.
DEFINITIONS
[0064] The term "treat," "treated," "treating" or "treatment"
includes the diminishment or alleviation of at least one symptom
associated or caused by the state, disorder or disease being
treated. In certain embodiments, the treatment comprises the
induction of a protein kinase-associated disorder, followed by the
activation of the compound of the invention, which would in turn
diminish or alleviate at least one symptom associated or caused by
the protein kinase-associated disorder being treated. For example,
treatment can be diminishment of one or several symptoms of a
disorder or complete eradication of a disorder.
[0065] The term "subject" is intended to include organisms, e.g.,
prokaryotes and eukaryotes, which are capable of suffering from or
afflicted with a disease, disorder or condition associated with the
activity of a protein kinase. Examples of subjects include mammals,
e.g., humans, dogs, cows, horses, pigs, sheep, goats, cats, mice,
rabbits, rats, and transgenic non-human animals. In certain
embodiments, the subject is a human, e.g., a human suffering from,
at risk of suffering from, or potentially capable of suffering from
cancer, transplant rejections, and autoimmune diseases, and for
other diseases or conditions described herein. In another
embodiment, the subject is a cell.
[0066] The language "protein kinase-modulating compound,"
"modulator of protein kinase" or "protein kinase inhibitor" refers
to compounds that modulate, e.g., inhibit, or otherwise alter, the
activity of a protein kinase. Examples of protein kinase-modulating
compounds include compounds of Formula I, as well as Table A, Table
B, Table C, Table D, Table E, and other examples as described
herein (including pharmaceutically acceptable salts thereof, as
well as enantiomers, stereoisomers, rotamers, tautomers,
diastereomers, atropisomers or racemates thereof).
[0067] Additionally, a method of the invention includes
administering to a subject an effective amount of a protein
kinase-modulating compound of the invention, e.g., protein
kinase-modulating compounds of Formula I, as well as Table A, Table
B, Table C, Table D, Table E, and other examples as described
herein (including pharmaceutically acceptable salts thereof, as
well as enantiomers, stereoisomers, rotamers, tautomers,
diastereomers, atropisomers or racemates thereof).
[0068] The term "alkyl" includes saturated aliphatic groups,
including straight-chain alkyl groups (e.g., methyl, ethyl, propyl,
butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc.),
branched-chain alkyl groups (isopropyl, tert-butyl, isobutyl,
etc.), cycloalkyl (alicyclic) groups (cyclopropyl, cyclopentyl,
cyclohexyl, cycloheptyl, cyclooctyl), alkyl substituted cycloalkyl
groups, and cycloalkyl substituted alkyl groups. The term "alkyl"
also includes alkenyl groups and alkynyl groups. Furthermore, the
expression "C.sub.x-C.sub.y-alkyl", wherein x is 1-5 and y is 2-10
indicates a particular alkyl group (straight- or branched-chain) of
a particular range of carbons. For example, the expression
C.sub.1-C.sub.4-alkyl includes, but is not limited to, methyl,
ethyl, propyl, butyl, isopropyl, tert-butyl and isobutyl. Moreover,
the term C.sub.3-6-cycloalkyl includes, but is not limited to,
cyclopropyl, cyclopentyl, and cyclohexyl. As discussed below, these
alkyl groups, as well as cycloalkyl groups, may be further
substituted.
[0069] The term "halo" as used herein means halogen, and includes
fluorine, chlorine, bromine, or iodine, especially fluorine and
chlorine.
[0070] The term alkyl further includes alkyl groups which can
further include oxygen, nitrogen, sulfur or phosphorous atoms
replacing one or more carbons of the hydrocarbon backbone. In an
embodiment, a straight chain or branched chain alkyl has 10 or
fewer carbon atoms in its backbone (e.g., C.sub.1-C.sub.10 for
straight chain, C.sub.3-C.sub.10 for branched chain), and more
preferably 6 or fewer carbons. Likewise, preferred cycloalkyls have
from 4-7 carbon atoms in their ring structure, and more preferably
have 5 or 6 carbons in the ring structure.
[0071] Moreover, alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl,
hexyl, etc.) include both "unsubstituted alkyl" and "substituted
alkyl", the latter of which refers to alkyl moieties having
substituents replacing a hydrogen on one or more carbons of the
hydrocarbon backbone, which allow the molecule to perform its
intended function.
[0072] The term "substituted" is intended to describe moieties
having substituents replacing a hydrogen on one or more atoms, e.g.
C, O or N, of a molecule. Such substituents can include, for
example, oxo, alkyl, alkoxy, alkenyl, alkynyl, halogen, hydroxyl,
alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,
phosphonato, phosphinato, amino (including alkyl amino,
dialkylamino, arylamino, diarylamino, and alkylarylamino),
acylamino (including alkylcarbonylamino, arylcarbonylamino,
carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio,
arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkylaryl, morpholino, phenol, benzyl, phenyl,
piperizine, cyclopentane, cyclohexane, pyridine, 5H-tetrazole,
triazole, piperidine, or an aromatic or heteroaromatic moiety, and
any combination thereof.
[0073] Further examples of substituents of the invention, which are
not intended to be limiting, include moieties selected from
straight or branched alkyl (preferably C.sub.1-C.sub.5), cycloalkyl
(preferably C.sub.3-C.sub.8), alkoxy (preferably C.sub.1-C.sub.6),
thioalkyl (preferably C.sub.1-C.sub.6), alkenyl (preferably
C.sub.2-C.sub.6), alkynyl (preferably C.sub.2-C.sub.6),
heterocyclic, carbocyclic, aryl (e.g., phenyl), aryloxy (e.g.,
phenoxy), aralkyl (e.g., benzyl), aryloxyalkyl (e.g.,
phenyloxyalkyl), arylacetamidoyl, alkylaryl, heteroaralkyl,
alkylcarbonyl and arylcarbonyl or other such acyl group,
heteroarylcarbonyl, or heteroaryl group, (CR'R'').sub.0-3NR'R''
(e.g., --NH.sub.2), (CR'R'').sub.0-3CN (e.g., --CN), --NO.sub.2,
halogen (e.g., --F, --Cl, --Br, or --I),
(CR'R'').sub.0-3C(halogen).sub.3 (e.g., --CF.sub.3),
(CR'R'').sub.0-3CH(halogen).sub.2,
(CR'R'').sub.0-3CH.sub.2(halogen), (CR'R'').sub.0-3CONR'R'',
(CR'R'').sub.0-3(CNH)NR'R'', (CR'R'').sub.0-3S(O).sub.1-2NR'R'',
(CR'R'').sub.0-3CHO, (CR'R'').sub.0-3O(CR'R'').sub.0-3H,
(CR'R'').sub.0-3S(O).sub.0-3R' (e.g., --SO.sub.3H, --OSO.sub.3H),
(CR'R'').sub.0-3O(CR'R'').sub.0-3H (e.g., --CH.sub.2OCH.sub.3 and
--OCH.sub.3), (CR'R'').sub.0-3S(CR'R'').sub.0-3H (e.g., --SH and
--SCH.sub.3), (CR'R'').sub.0-3OH (e.g., --OH),
(CR'R'').sub.0-3COR', (CR'R'').sub.0-3 (substituted or
unsubstituted phenyl), (CR'R'').sub.0-3(C.sub.3-C.sub.8
cycloalkyl), (CR'R'').sub.0-3CO.sub.2R' (e.g., --CO.sub.2H), or
(CR'R'').sub.0-3OR' group, or the side chain of any naturally
occurring amino acid; wherein R' and R'' are each independently
hydrogen, a C.sub.1-C.sub.5 alkyl, C.sub.2-C.sub.5 alkenyl,
C.sub.2-C.sub.5 alkynyl, or aryl group. Such substituents can
include, for example, halogen, hydroxyl, alkylcarbonyloxy,
arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy,
carboxylate, alkylcarbonyl, alkoxycarbonyl, aminocarbonyl,
alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato,
cyano, amino (including alkyl amino, dialkylamino, arylamino,
diarylamino, and alkylarylamino), acylamino (including
alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),
amidino, imino, oxime, sulfhydryl, alkylthio, arylthio,
thiocarboxylate, sulfates, sulfonato, sulfamoyl, sulfonamido,
nitro, trifluoromethyl, cyano, azido, heterocyclyl, or an aromatic
or heteroaromatic moiety, and any combination thereof. In certain
embodiments, a carbonyl moiety (C.dbd.O) may be further derivatized
with an oxime moiety, e.g., an aldehyde moiety may be derivatized
as its oxime (--C.dbd.N--OH) analog. It will be understood by those
skilled in the art that the moieties substituted on the hydrocarbon
chain can themselves be substituted, if appropriate. Cycloalkyls
can be further substituted, e.g., with the substituents described
above. An "aralkyl" moiety is an alkyl substituted with an aryl
(e.g., phenylmethyl (i.e., benzyl)).
[0074] The term "alkenyl" includes unsaturated aliphatic groups
analogous in length and possible substitution to the alkyls
described above, but which contain at least one double bond.
[0075] For example, the term "alkenyl" includes straight-chain
alkenyl groups (e.g., ethenyl, propenyl, butenyl, pentenyl,
hexenyl, heptenyl, octenyl, nonenyl, decenyl, etc.), branched-chain
alkenyl groups, cycloalkenyl (alicyclic) groups (cyclopropenyl,
cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl), alkyl or
alkenyl substituted cycloalkenyl groups, and cycloalkyl or
cycloalkenyl substituted alkenyl groups. The term alkenyl further
includes alkenyl groups that include oxygen, nitrogen, sulfur or
phosphorous atoms replacing one or more carbons of the hydrocarbon
backbone. In certain embodiments, a straight chain or branched
chain alkenyl group has 6 or fewer carbon atoms in its backbone
(e.g., C.sub.2-C.sub.6 for straight chain, C.sub.3-C.sub.6 for
branched chain). Likewise, cycloalkenyl groups may have from 3-8
carbon atoms in their ring structure, and more preferably have 5 or
6 carbons in the ring structure. The term C.sub.2-C.sub.6 includes
alkenyl groups containing 2 to 6 carbon atoms.
[0076] Moreover, the term alkenyl includes both "unsubstituted
alkenyls" and "substituted alkenyls", the latter of which refers to
alkenyl moieties having substituents replacing a hydrogen on one or
more carbons of the hydrocarbon backbone. Such substituents can
include, for example, alkyl groups, alkynyl groups, halogens,
hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,
phosphonato, phosphinato, cyano, amino (including alkyl amino,
dialkylamino, arylamino, diarylamino, and alkylarylamino),
acylamino (including alkylcarbonylamino, arylcarbonylamino,
carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio,
arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkylaryl, or an aromatic or heteroaromatic
moiety.
[0077] The term "alkynyl" includes unsaturated aliphatic groups
analogous in length and possible substitution to the alkyls
described above, but which contain at least one triple bond.
[0078] For example, the term "alkynyl" includes straight-chain
alkynyl groups (e.g., ethynyl, propynyl, butynyl, pentynyl,
hexynyl, heptynyl, octynyl, nonynyl, decynyl, etc.), branched-chain
alkynyl groups, and cycloalkyl or cycloalkenyl substituted alkynyl
groups. The term alkynyl further includes alkynyl groups that
include oxygen, nitrogen, sulfur or phosphorous atoms replacing one
or more carbons of the hydrocarbon backbone. In certain
embodiments, a straight chain or branched chain alkynyl group has 6
or fewer carbon atoms in its backbone (e.g., C.sub.2-C.sub.6 for
straight chain, C.sub.3-C.sub.6 for branched chain). The term
C.sub.2-C.sub.6 includes alkynyl groups containing 2 to 6 carbon
atoms.
[0079] Moreover, the term alkynyl includes both "unsubstituted
alkynyls" and "substituted alkynyls", the latter of which refers to
alkynyl moieties having substituents replacing a hydrogen on one or
more carbons of the hydrocarbon backbone. Such substituents can
include, for example, alkyl groups, alkynyl groups, halogens,
hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,
phosphonato, phosphinato, cyano, amino (including alkyl amino,
dialkylamino, arylamino, diarylamino, and alkylarylamino),
acylamino (including alkylcarbonylamino, arylcarbonylamino,
carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio,
arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkylaryl, or an aromatic or heteroaromatic
moiety.
[0080] The term "amine" or "amino" should be understood as being
broadly applied to both a molecule, or a moiety or functional
group, as generally understood in the art, and may be primary,
secondary, or tertiary. The term "amine" or "amino" includes
compounds where a nitrogen atom is covalently bonded to at least
one carbon, hydrogen or heteroatom. The terms include, for example,
but are not limited to, "alkylamino," "arylamino," "diarylamino,"
"alkylarylamino," "alkylaminoaryl," "arylaminoalkyl,"
"alkaminoalkyl," "amide," "amido," and "aminocarbonyl." The term
"alkyl amino" comprises groups and compounds wherein the nitrogen
is bound to at least one additional alkyl group. The term "dialkyl
amino" includes groups wherein the nitrogen atom is bound to at
least two additional alkyl groups. The term "arylamino" and
"diarylamino" include groups wherein the nitrogen is bound to at
least one or two aryl groups, respectively. The term
"alkylarylamino," "alkylaminoaryl" or "arylaminoalkyl" refers to an
amino group which is bound to at least one alkyl group and at least
one aryl group. The term "alkaminoalkyl" refers to an alkyl,
alkenyl, or alkynyl group bound to a nitrogen atom which is also
bound to an alkyl group.
[0081] The term "amide," "amido" or "aminocarbonyl" includes
compounds or moieties which contain a nitrogen atom which is bound
to the carbon of a carbonyl or a thiocarbonyl group. The term
includes "alkaminocarbonyl" or "alkylaminocarbonyl" groups which
include alkyl, alkenyl, aryl or alkynyl groups bound to an amino
group bound to a carbonyl group. It includes arylaminocarbonyl and
arylcarbonylamino groups which include aryl or heteroaryl moieties
bound to an amino group which is bound to the carbon of a carbonyl
or thiocarbonyl group. The terms "alkylaminocarbonyl,"
"alkenylaminocarbonyl," "alkynylaminocarbonyl,"
"arylaminocarbonyl," "alkylcarbonylamino," "alkenylcarbonylamino,"
"alkynylcarbonylamino," and "arylcarbonylamino" are included in
term "amide." Amides also include urea groups (aminocarbonylamino)
and carbamates (oxycarbonylamino).
[0082] The term "aryl" includes groups, including 5- and 6-membered
single-ring aromatic groups that may include from zero to four
heteroatoms, for example, phenyl, pyrrole, furan, thiophene,
thiazole, isothiaozole, imidazole, triazole, tetrazole, pyrazole,
oxazole, isoxazole, pyridine, pyrazine, pyridazine, and pyrimidine,
and the like. Furthermore, the term "aryl" includes multicyclic
aryl groups, e.g., tricyclic, bicyclic, e.g., naphthalene,
benzoxazole, benzodioxazole, benzothiazole, benzoimidazole,
benzothiophene, methylenedioxyphenyl, quinoline, isoquinoline,
anthryl, phenanthryl, napthridine, indole, benzofuran, purine,
benzofuran, deazapurine, or indolizine. Those aryl groups having
heteroatoms in the ring structure may also be referred to as "aryl
heterocycles", "heterocycles," "heteroaryls" or "heteroaromatics."
The aromatic ring can be substituted at one or more ring positions
with such substituents as described above, as for example, alkyl,
halogen, hydroxyl, alkoxy, alkylcarbonyloxy, arylcarbonyloxy,
alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,
alkylaminoacarbonyl, aralkylaminocarbonyl, alkenylaminocarbonyl,
alkylcarbonyl, arylcarbonyl, aralkylcarbonyl, alkenylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, phosphate,
phosphonato, phosphinato, cyano, amino (including alkyl amino,
dialkylamino, arylamino, diarylamino, and alkylarylamino),
acylamino (including alkylcarbonylamino, arylcarbonylamino,
carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio,
arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.
Aryl groups can also be fused or bridged with alicyclic or
heterocyclic rings which are not aromatic so as to form a polycycle
(e.g., tetralin).
[0083] The term heteroaryl, as used herein, represents a stable
monocyclic or bicyclic ring of up to 7 atoms in each ring, wherein
at least one ring is aromatic and contains from 1 to 4 heteroatoms
selected from the group consisting of O, N and S. Heteroaryl groups
within the scope of this definition include but are not limited to:
acridinyl, carbazolyl, cinnolinyl, quinoxalinyl, pyrrazolyl,
indolyl, benzotriazolyl, furanyl, thienyl, benzothienyl,
benzofuranyl, quinolinyl, isoquinolinyl, oxazolyl, isoxazolyl,
indolyl, pyrazinyl, pyridazinyl, pyridinyl, pyrimidinyl, pyrrolyl,
tetrahydroquinoline. As with the definition of heterocycle below,
"heteroaryl" is also understood to include the N-oxide derivative
of any nitrogen-containing heteroaryl. In cases where the
heteroaryl substituent is bicyclic and one ring is non-aromatic or
contains no heteroatoms, it is understood that attachment is via
the aromatic ring or via the heteroatom containing ring,
respectively.
[0084] The term "heterocycle" or "heterocyclyl" as used herein is
intended to mean a 5- to 10-membered aromatic or nonaromatic
heterocycle containing from 1 to 4 heteroatoms selected from the
group consisting of O, N and S, and includes bicyclic groups.
"Heterocyclyl" therefore includes the above mentioned heteroaryls,
as well as dihydro and tetrathydro analogs thereof. Further
examples of "heterocyclyl" include, but are not limited to the
following: benzoimidazolyl, benzofuranyl, benzofurazanyl,
benzopyrazolyl, benzotriazolyl, benzothiophenyl, benzoxazolyl,
carbazolyl, carbolinyl, cinnolinyl, furanyl, imidazolyl, indolinyl,
indolyl, indolazinyl, indazolyl, isobenzofuranyl, isoindolyl,
isoquinolyl, isothiazolyl, isoxazolyl, naphthpyridinyl,
oxadiazolyl, oxazolyl, oxazoline, isoxazoline, oxetanyl, pyranyl,
pyrazinyl, pyrazolyl, pyridazinyl, pyridopyridinyl, pyridazinyl,
pyridyl, pyrimidyl, pyrrolyl, quinazolinyl, quinolyl, quinoxalinyl,
tetrahydropyranyl, tetrazolyl, tetrazolopyridyl, thiadiazolyl,
thiazolyl, thienyl, triazolyl, azetidinyl, 1,4-dioxanyl,
hexahydroazepinyl, piperazinyl, piperidinyl, pyridin-2-onyl,
pyrrolidinyl, morpholinyl, thiomorpholinyl, dihydrobenzoimidazolyl,
dihydrobenzofuranyl, dihydrobenzothiophenyl, dihydrobenzoxazolyl,
dihydrofuranyl, dihydroimidazolyl, dihydroindolyl,
dihydroisooxazolyl, dihydroisothiazolyl, dihydrooxadiazolyl,
dihydrooxazolyl, dihydropyrazinyl, dihydropyrazolyl,
dihydropyridinyl, dihydropyrimidinyl, dihydropyrrolyl,
dihydroquinolinyl, dihydrotetrazolyl, dihydrothiadiazolyl,
dihydrothiazolyl, dihydrothienyl, dihydrotriazolyl,
dihydroazetidinyl, methylenedioxybenzoyl, tetrahydrofuranyl, and
tetrahydrothienyl, and N-oxides thereof. Attachment of a
heterocyclyl substituent can occur via a carbon atom or via a
heteroatom.
[0085] The term "acyl" includes compounds and moieties which
contain the acyl radical (CH.sub.3CO--) or a carbonyl group. The
term "substituted acyl" includes acyl groups where one or more of
the hydrogen atoms are replaced by for example, alkyl groups,
alkynyl groups, halogens, hydroxyl, alkylcarbonyloxy,
arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy,
carboxylate, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl,
aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl,
alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato,
cyano, amino (including alkyl amino, dialkylamino, arylamino,
diarylamino, and alkylarylamino), acylamino (including
alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),
amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,
sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,
trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an
aromatic or heteroaromatic moiety.
[0086] The term "acylamino" includes moieties wherein an acyl
moiety is bonded to an amino group. For example, the term includes
alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido
groups.
[0087] The term "alkoxy" includes substituted and unsubstituted
alkyl, alkenyl, and alkynyl groups covalently linked to an oxygen
atom. Examples of alkoxy groups include methoxy, ethoxy,
isopropyloxy, propoxy, butoxy, and pentoxy groups and may include
cyclic groups such as cyclopentoxy. Examples of substituted alkoxy
groups include halogenated alkoxy groups. The alkoxy groups can be
substituted with groups such as alkenyl, alkynyl, halogen,
hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,
phosphonato, phosphinato, cyano, amino (including alkyl amino,
dialkylamino, arylamino, diarylamino, and alkylarylamino),
acylamino (including alkylcarbonylamino, arylcarbonylamino,
carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio,
arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moieties.
Examples of halogen substituted alkoxy groups include, but are not
limited to, fluoromethoxy, difluoromethoxy, trifluoromethoxy,
chloromethoxy, dichloromethoxy, trichloromethoxy, etc.
[0088] The term "carbonyl" or "carboxy" includes compounds and
moieties which contain a carbon connected with a double bond to an
oxygen atom, and tautomeric forms thereof. Examples of moieties
that contain a carbonyl include aldehydes, ketones, carboxylic
acids, amides, esters, anhydrides, etc. The term "carboxy moiety"
or "carbonyl moiety" refers to groups such as "alkylcarbonyl"
groups wherein an alkyl group is covalently bound to a carbonyl
group, "alkenylcarbonyl" groups wherein an alkenyl group is
covalently bound to a carbonyl group, "alkynylcarbonyl" groups
wherein an alkynyl group is covalently bound to a carbonyl group,
"arylcarbonyl" groups wherein an aryl group is covalently attached
to the carbonyl group. Furthermore, the term also refers to groups
wherein one or more heteroatoms are covalently bonded to the
carbonyl moiety. For example, the term includes moieties such as,
for example, aminocarbonyl moieties, (wherein a nitrogen atom is
bound to the carbon of the carbonyl group, e.g., an amide),
aminocarbonyloxy moieties, wherein an oxygen and a nitrogen atom
are both bond to the carbon of the carbonyl group (e.g., also
referred to as a "carbamate"). Furthermore, aminocarbonylamino
groups (e.g., ureas) are also include as well as other combinations
of carbonyl groups bound to heteroatoms (e.g., nitrogen, oxygen,
sulfur, etc. as well as carbon atoms). Furthermore, the heteroatom
can be further substituted with one or more alkyl, alkenyl,
alkynyl, aryl, aralkyl, acyl, etc. moieties.
[0089] The term "thiocarbonyl" or "thiocarboxy" includes compounds
and moieties which contain a carbon connected with a double bond to
a sulfur atom. The term "thiocarbonyl moiety" includes moieties
that are analogous to carbonyl moieties. For example,
"thiocarbonyl" moieties include aminothiocarbonyl, wherein an amino
group is bound to the carbon atom of the thiocarbonyl group,
furthermore other thiocarbonyl moieties include, oxythiocarbonyls
(oxygen bound to the carbon atom), aminothiocarbonylamino groups,
etc.
[0090] The term "ether" includes compounds or moieties that contain
an oxygen bonded to two different carbon atoms or heteroatoms. For
example, the term includes "alkoxyalkyl" which refers to an alkyl,
alkenyl, or alkynyl group covalently bonded to an oxygen atom that
is covalently bonded to another alkyl group.
[0091] The term "ester" includes compounds and moieties that
contain a carbon or a heteroatom bound to an oxygen atom that is
bonded to the carbon of a carbonyl group. The term "ester" includes
alkoxycarboxy groups such as methoxycarbonyl, ethoxycarbonyl,
propoxycarbonyl, butoxycarbonyl, pentoxycarbonyl, etc. The alkyl,
alkenyl, or alkynyl groups are as defined above.
[0092] The term "thioether" includes compounds and moieties which
contain a sulfur atom bonded to two different carbon or hetero
atoms. Examples of thioethers include, but are not limited to
alkthioalkyls, alkthioalkenyls, and alkthioalkynyls. The term
"alkthioalkyls" include compounds with an alkyl, alkenyl, or
alkynyl group bonded to a sulfur atom that is bonded to an alkyl
group. Similarly, the term "alkthioalkenyls" and alkthioalkynyls"
refer to compounds or moieties wherein an alkyl, alkenyl, or
alkynyl group is bonded to a sulfur atom which is covalently bonded
to an alkynyl group.
[0093] The term "hydroxy" or "hydroxyl" includes groups with an
--OH or --O.sup.-.
[0094] The term "halogen" includes fluorine, bromine, chlorine,
iodine, etc. The term "perhalogenated" generally refers to a moiety
wherein all hydrogens are replaced by halogen atoms.
[0095] The terms "polycyclyl" or "polycyclic radical" include
moieties with two or more rings (e.g., cycloalkyls, cycloalkenyls,
cycloalkynyls, aryls and/or heterocyclyls) in which two or more
carbons are common to two adjoining rings, e.g., the rings are
"fused rings". Rings that are joined through non-adjacent atoms are
termed "bridged" rings. Each of the rings of the polycycle can be
substituted with such substituents as described above, as for
example, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy,
alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,
alkoxycarbonyl, alkylaminoacarbonyl, aralkylaminocarbonyl,
alkenylaminocarbonyl, alkylcarbonyl, arylcarbonyl, aralkylcarbonyl,
alkenylcarbonyl, aminocarbonyl, alkylthiocarbonyl, alkoxyl,
phosphate, phosphonato, phosphinato, cyano, amino (including alkyl
amino, dialkylamino, arylamino, diarylamino, and alkylarylamino),
acylamino (including alkylcarbonylamino, arylcarbonylamino,
carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio,
arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkyl, alkylaryl, or an aromatic or heteroaromatic
moiety.
[0096] The term "heteroatom" includes atoms of any element other
than carbon or hydrogen. Preferred heteroatoms are nitrogen,
oxygen, sulfur and phosphorus.
[0097] Additionally, the phrase "any combination thereof" implies
that any number of the listed functional groups and molecules may
be combined to create a larger molecular architecture. For example,
the terms "phenyl," "carbonyl" (or ".dbd.O"), "--O--," "--OH," and
C.sub.1-6 (i.e., --CH.sub.3 and --CH.sub.2CH.sub.2CH.sub.2--) can
be combined to form a 3-methoxy-4-propoxybenzoic acid substituent.
It is to be understood that when combining functional groups and
molecules to create a larger molecular architecture, hydrogens can
be removed or added, as required to satisfy the valence of each
atom.
[0098] It is to be understood that all of the compounds of the
invention described above will further include bonds between
adjacent atoms and/or hydrogens as required to satisfy the valence
of each atom. That is, bonds and/or hydrogen atoms are added to
provide the following number of total bonds to each of the
following types of atoms: carbon: four bonds; nitrogen: three
bonds; oxygen: two bonds; and sulfur: two-six bonds.
[0099] It will be noted that the structures of some of the
compounds of this invention include asymmetric carbon atoms. It is
to be understood accordingly that the isomers arising from such
asymmetry (e.g., all enantiomers, stereoisomers, rotamers,
tautomers, diastereomers, or racemates) are included within the
scope of this invention. Such isomers can be obtained in
substantially pure form by classical separation techniques and by
stereochemically controlled synthesis. Furthermore, the structures
and other compounds and moieties discussed in this application also
include all tautomers thereof. Compounds described herein may be
obtained through art recognized synthesis strategies.
[0100] It will also be noted that the substituents of some of the
compounds of this invention include isomeric cyclic structures. It
is to be understood accordingly that constitutional isomers of
particular substituents are included within the scope of this
invention, unless indicated otherwise. For example, the term
"tetrazole" includes tetrazole, 2H-tetrazole, 3H-tetrazole,
4H-tetrazole and 5H-tetrazole.
Use in Cancer, Transplant Rejections, and Autoimmune Diseases
[0101] The compounds of the present invention have valuable
pharmacological properties and are useful in the treatment of
diseases. In certain embodiments, compounds of the invention are
useful in the treatment of a proliferative disease, or cancer.
[0102] A proliferative disease is mainly a tumor disease (or
cancer) (and/or any metastases). The inventive compounds are
particularly useful for treating a tumor which is a breast cancer,
genitourinary cancer, lung cancer, gastrointestinal cancer,
epidermoid cancer, melanoma, ovarian cancer, pancreas cancer,
neuroblastoma, head and/or neck cancer or bladder cancer, or in a
broader sense renal, brain or gastric cancer; in particular (i) a
breast tumor; an epidermoid tumor, such as an epidermoid head
and/or neck tumor or a mouth tumor; a lung tumor, for example a
small cell or non-small cell lung tumor; a gastrointestinal tumor,
for example, a colorectal tumor; or a genitourinary tumor, for
example, a prostate tumor (especially a hormone-refractory prostate
tumor); or (ii) a proliferative disease that is refractory to the
treatment with other chemotherapeutics; or (iii) a tumor that is
refractory to treatment with other chemotherapeutics due to
multidrug resistance.
[0103] In a broader sense of the invention, a proliferative disease
may furthermore be a hyperproliferative condition such as
leukemias, hyperplasias, fibrosis (especially pulmonary, but also
other types of fibrosis, such as renal fibrosis), angiogenesis,
psoriasis, atherosclerosis and smooth muscle proliferation in the
blood vessels, such as stenosis or restenosis following
angioplasty.
[0104] Where a tumor, a tumor disease, a carcinoma or a cancer are
mentioned, also metastasis in the original organ or tissue and/or
in any other location are implied alternatively or in addition,
whatever the location of the tumor and/or metastasis.
[0105] The inventive compound is selectively toxic or more toxic to
rapidly proliferating cells than to normal cells, particularly in
human cancer cells, e.g., cancerous tumors, the compound has
significant antiproliferative effects and promotes differentiation,
e.g., cell cycle arrest and apoptosis.
[0106] In other certain embodiments, compounds of the invention are
useful in the treatment of transplant rejections. Examples of
transplant rejections that may be treated by the compounds of the
invention include, but are not limited to, graft versus host
disease, rejection related to xeno transplantation, rejection
related to organ transplant, rejection related to acute transplant,
heterograft or homograft rejection and ischemic or reperfusion
injury incurred during organ transplantation.
[0107] In still other certain embodiments, compounds of the
invention are useful in the treatment of autoimmune diseases.
Examples of autoimmune diseases to be treated by the compounds of
the invention include, but are not limited to, autoimmune hemolytic
anemia, autoimmune neonatal thrombocytopenia, idiopathic
thrombocytopenia purpura, autoimmunocytopenia, hemolytic anemia,
antiphospholipid syndrome, dermatitis, allergic encephalomyelitis,
myocarditis, relapsing polychondritis, rheumatic heart disease,
glomerulonephritis, multiple sclerosis, neuritis, uveitis
ophthalmia, polyendocrinopathies, purpura, Reiter's Disease,
Stiff-Man Syndrome, autoimmune pulmonary inflammation, autism,
Guillain-Barre Syndrome, insulin dependent diabetes mellitis,
autoimmune inflammatory eye, autoimmune thyroiditis,
hypothyroidism, systemic lupus erhythematosus, Goodpasture's
syndrome, Pemphigus, Receptor autoimmunities, autoimmune hemolytic
anemia, autoimmune thrombocytopenic purpura, rheumatoid arthritis,
mixed connective tissue disease, polymyositis/dermatomyositis,
pernicious anemia, idiopathic Addison's disease, infertility,
glomerulonephritis, bullous pemphigoid, Sjogren's syndrome,
diabetes millitus, adrenergic drug resistance, chronic active
hepatitis, primary biliary cirrhosis, vitiligo, vasculitis,
post-MI, cardiotomy syndrome, urticaria, atopic dermatitis, asthma,
inflammatory myopathies, chronic active hepatitis, primary biliary
cirrhosis and T-cell mediated hypersensitivity diseases.
[0108] The term "use" includes any one or more of the following
embodiments of the invention, respectively: the use in the
treatment of protein kinase-associated disorders; the use for the
manufacture of pharmaceutical compositions for use in the treatment
of these diseases, e.g., in the manufacture of a medicament;
methods of use of compounds of the invention in the treatment of
these diseases; pharmaceutical preparations having compounds of the
invention for the treatment of these diseases; and compounds of the
invention for use in the treatment of these diseases; as
appropriate and expedient, if not stated otherwise. In particular,
diseases to be treated and are thus preferred for use of a compound
of the present invention are selected from cancer, transplant
rejections, or autoimmune diseases, as well as those diseases that
depend on the activity of protein kinases. The term "use" further
includes embodiments of compositions herein which bind to a protein
kinase sufficiently to serve as tracers or labels, so that when
coupled to a fluor or tag, or made radioactive, can be used as a
research reagent or as a diagnostic or an imaging agent.
Assays
[0109] The inhibition of protein kinase activity by the compounds
of the invention may be measured using a number of assays available
in the art. Examples of such assays are described in the
Exemplification section below.
Pharmaceutical Compositions
[0110] The language "effective amount" of the compound is that
amount necessary or sufficient to treat or prevent a protein
kinase-associated disorder, e.g. prevent the various morphological
and somatic symptoms of a protein kinase-associated disorder,
and/or a disease or condition described herein. In an example, an
effective amount of the compound of the invention is the amount
sufficient to treat a protein kinase-associated disorder in a
subject. The effective amount can vary depending on such factors as
the size and weight of the subject, the type of illness, or the
particular compound of the invention. For example, the choice of
the compound of the invention can affect what constitutes an
"effective amount." One of ordinary skill in the art would be able
to study the factors contained herein and make the determination
regarding the effective amount of the compounds of the invention
without undue experimentation.
[0111] The regimen of administration can affect what constitutes an
effective amount. The compound of the invention can be administered
to the subject either prior to or after the onset of a protein
kinase-associated disorder. Further, several divided dosages, as
well as staggered dosages, can be administered daily or
sequentially, or the dose can be continuously infused, or can be a
bolus injection. Further, the dosages of the compound(s) of the
invention can be proportionally increased or decreased as indicated
by the exigencies of the therapeutic or prophylactic situation.
[0112] Compounds of the invention may be used in the treatment of
states, disorders or diseases as described herein, or for the
manufacture of pharmaceutical compositions for use in the treatment
of these diseases. Methods of use of compounds of the present
invention in the treatment of these diseases, or pharmaceutical
preparations having compounds of the present invention for the
treatment of these diseases.
[0113] The language "pharmaceutical composition" includes
preparations suitable for administration to mammals, e.g., humans.
When the compounds of the present invention are administered as
pharmaceuticals to mammals, e.g., humans, they can be given per se
or as a pharmaceutical composition containing, for example, 0.1 to
99.5% (more preferably, 0.5 to 90%) of active ingredient in
combination with a pharmaceutically acceptable carrier.
[0114] The phrase "pharmaceutically acceptable carrier" is art
recognized and includes a pharmaceutically acceptable material,
composition or vehicle, suitable for administering compounds of the
present invention to mammals. The carriers include liquid or solid
filler, diluent, excipient, solvent or encapsulating material,
involved in carrying or transporting the subject agent from one
organ, or portion of the body, to another organ, or portion of the
body. Each carrier must be "acceptable" in the sense of being
compatible with the other ingredients of the formulation and not
injurious to the patient. Some examples of materials which can
serve as pharmaceutically acceptable carriers include: sugars, such
as lactose, glucose and sucrose; starches, such as corn starch and
potato starch; cellulose, and its derivatives, such as sodium
carboxymethyl cellulose, ethyl cellulose and cellulose acetate;
powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa
butter and suppository waxes; oils, such as peanut oil, cottonseed
oil, safflower oil, sesame oil, olive oil, corn oil and soybean
oil; glycols, such as propylene glycol; polyols, such as glycerin,
sorbitol, mannitol and polyethylene glycol; esters, such as ethyl
oleate and ethyl laurate; agar; buffering agents, such as magnesium
hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water;
isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer
solutions; and other non-toxic compatible substances employed in
pharmaceutical formulations.
[0115] Wetting agents, emulsifiers and lubricants, such as sodium
lauryl sulfate and magnesium stearate, as well as coloring agents,
release agents, coating agents, sweetening, flavoring and perfuming
agents, preservatives and antioxidants can also be present in the
compositions.
[0116] Examples of pharmaceutically acceptable antioxidants
include: water soluble antioxidants, such as ascorbic acid,
cysteine hydrochloride, sodium bisulfate, sodium metabisulfite,
sodium sulfite and the like; oil-soluble antioxidants, such as
ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated
hydroxytoluene (BHT), lecithin, propyl gallate, .alpha.-tocopherol,
and the like; and metal chelating agents, such as citric acid,
ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid,
phosphoric acid, and the like.
[0117] Formulations of the present invention include those suitable
for oral, nasal, topical, buccal, sublingual, rectal, vaginal
and/or parenteral administration. The formulations may conveniently
be presented in unit dosage form and may be prepared by any methods
well known in the art of pharmacy. The amount of active ingredient
that can be combined with a carrier material to produce a single
dosage form will generally be that amount of the compound that
produces a therapeutic effect. Generally, out of one hundred
percent, this amount will range from about 1 percent to about
ninety-nine percent of active ingredient, preferably from about 5
percent to about 70 percent, most preferably from about 10 percent
to about 30 percent.
[0118] Methods of preparing these formulations or compositions
include the step of bringing into association a compound of the
present invention with the carrier and, optionally, one or more
accessory ingredients. In general, the formulations are prepared by
uniformly and intimately bringing into association a compound of
the present invention with liquid carriers, or finely divided solid
carriers, or both, and then, if necessary, shaping the product.
[0119] Formulations of the invention suitable for oral
administration may be in the form of capsules, cachets, pills,
tablets, lozenges (using a flavored basis, usually sucrose and
acacia or tragacanth), powders, granules, or as a solution or a
suspension in an aqueous or non-aqueous liquid, or as an
oil-in-water or water-in-oil liquid emulsion, or as an elixir or
syrup, or as pastilles (using an inert base, such as gelatin and
glycerin, or sucrose and acacia) and/or as mouth washes and the
like, each containing a predetermined amount of a compound of the
present invention as an active ingredient. A compound of the
present invention may also be administered as a bolus, electuary or
paste.
[0120] In solid dosage forms of the invention for oral
administration (capsules, tablets, pills, dragees, powders,
granules and the like), the active ingredient is mixed with one or
more pharmaceutically acceptable carriers, such as sodium citrate
or dicalcium phosphate, and/or any of the following: fillers or
extenders, such as starches, lactose, sucrose, glucose, mannitol,
and/or silicic acid; binders, such as, for example,
carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone,
sucrose and/or acacia; humectants, such as glycerol; disintegrating
agents, such as agar-agar, calcium carbonate, potato or tapioca
starch, alginic acid, certain silicates, and sodium carbonate;
solution retarding agents, such as paraffin; absorption
accelerators, such as quaternary ammonium compounds; wetting
agents, such as, for example, cetyl alcohol and glycerol
monostearate; absorbents, such as kaolin and bentonite clay;
lubricants, such a talc, calcium stearate, magnesium stearate,
solid polyethylene glycols, sodium lauryl sulfate, and mixtures
thereof; and coloring agents. In the case of capsules, tablets and
pills, the pharmaceutical compositions may also comprise buffering
agents. Solid compositions of a similar type may also be employed
as fillers in soft and hard-filled gelatin capsules using such
excipients as lactose or milk sugars, as well as high molecular
weight polyethylene glycols and the like.
[0121] A tablet may be made by compression or molding, optionally
with one or more accessory ingredients. Compressed tablets may be
prepared using binder (for example, gelatin or hydroxypropylmethyl
cellulose), lubricant, inert diluent, preservative, disintegrant
(for example, sodium starch glycolate or cross-linked sodium
carboxymethyl cellulose), surface-active or dispersing agent.
Molded tablets may be made by molding in a suitable machine a
mixture of the powdered compound moistened with an inert liquid
diluent.
[0122] The tablets, and other solid dosage forms of the
pharmaceutical compositions of the present invention, such as
dragees, capsules, pills and granules, may optionally be scored or
prepared with coatings and shells, such as enteric coatings and
other coatings well known in the pharmaceutical-formulating art.
They may also be formulated so as to provide slow or controlled
release of the active ingredient therein using, for example,
hydroxypropylmethyl cellulose in varying proportions to provide the
desired release profile, other polymer matrices, liposomes and/or
microspheres. They may be sterilized by, for example, filtration
through a bacteria-retaining filter, or by incorporating
sterilizing agents in the form of sterile solid compositions that
can be dissolved in sterile water, or some other sterile injectable
medium immediately before use. These compositions may also
optionally contain opacifying agents and may be of a composition
that they release the active ingredient(s) only, or preferentially,
in a certain portion of the gastrointestinal tract, optionally, in
a delayed manner. Examples of embedding compositions that can be
used include polymeric substances and waxes. The active ingredient
can also be in micro-encapsulated form, if appropriate, with one or
more of the above-described excipients.
[0123] Liquid dosage forms for oral administration of the compounds
of the invention include pharmaceutically acceptable emulsions,
microemulsions, solutions, suspensions, syrups and elixirs. In
addition to the active ingredient, the liquid dosage forms may
contain inert diluent commonly used in the art, such as, for
example, water or other solvents, solubilizing agents and
emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl
carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate,
propylene glycol, 1,3-butylene glycol, oils (in particular,
cottonseed, groundnut, corn, germ, olive, castor and sesame oils),
glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty
acid esters of sorbitan, and mixtures thereof.
[0124] Besides inert diluents, the oral compositions can also
include adjuvants such as wetting agents, emulsifying and
suspending agents, sweetening, flavoring, coloring, perfuming and
preservative agents.
[0125] Suspensions, in addition to the active compounds, may
contain suspending agents as, for example, ethoxylated isostearyl
alcohols, polyoxyethylene sorbitol and sorbitan esters,
microcrystalline cellulose, aluminum metahydroxide, bentonite,
agar-agar and tragacanth, and mixtures thereof.
[0126] Formulations of the pharmaceutical compositions of the
invention for rectal or vaginal administration may be presented as
a suppository, which may be prepared by mixing one or more
compounds of the invention with one or more suitable nonirritating
excipients or carriers comprising, for example, cocoa butter,
polyethylene glycol, a suppository wax or a salicylate, and which
is solid at room temperature, but liquid at body temperature and,
therefore, will melt in the rectum or vaginal cavity and release
the active compound.
[0127] Formulations of the present invention which are suitable for
vaginal administration also include pessaries, tampons, creams,
gels, pastes, foams or spray formulations containing such carriers
as are known in the art to be appropriate.
[0128] Dosage forms for the topical or transdermal administration
of a compound of this invention include powders, sprays, ointments,
pastes, creams, lotions, gels, solutions, patches and inhalants.
The active compound may be mixed under sterile conditions with a
pharmaceutically acceptable carrier, and with any preservatives,
buffers, or propellants that may be required.
[0129] The ointments, pastes, creams and gels may contain, in
addition to an active compound of this invention, excipients, such
as animal and vegetable fats, oils, waxes, paraffins, starch,
tragacanth, cellulose derivatives, polyethylene glycols, silicones,
bentonites, silicic acid, talc and zinc oxide, or mixtures
thereof.
[0130] Powders and sprays can contain, in addition to a compound of
this invention, excipients such as lactose, talc, silicic acid,
aluminum hydroxide, calcium silicates and polyamide powder, or
mixtures of these substances. Sprays can additionally contain
customary propellants, such as chlorofluorohydrocarbons and
volatile unsubstituted hydrocarbons, such as butane and
propane.
[0131] Transdermal patches have the added advantage of providing
controlled delivery of a compound of the present invention to the
body. Such dosage forms can be made by dissolving or dispersing the
compound in the proper medium. Absorption enhancers can also be
used to increase the flux of the compound across the skin. The rate
of such flux can be controlled by either providing a rate
controlling membrane or dispersing the active compound in a polymer
matrix or gel.
[0132] Ophthalmic formulations, eye ointments, powders, solutions
and the like, are also contemplated as being within the scope of
this invention.
[0133] Pharmaceutical compositions of this invention suitable for
parenteral administration comprise one or more compounds of the
invention in combination with one or more pharmaceutically
acceptable sterile isotonic aqueous or nonaqueous solutions,
dispersions, suspensions or emulsions, or sterile powders which may
be reconstituted into sterile injectable solutions or dispersions
just prior to use, which may contain antioxidants, buffers,
bacteriostats, solutes which render the formulation isotonic with
the blood of the intended recipient or suspending or thickening
agents.
[0134] Examples of suitable aqueous and nonaqueous carriers that
may be employed in the pharmaceutical compositions of the invention
include water, ethanol, polyols (such as glycerol, propylene
glycol, polyethylene glycol, and the like), and suitable mixtures
thereof, vegetable oils, such as olive oil, and injectable organic
esters, such as ethyl oleate. Proper fluidity can be maintained,
for example, by the use of coating materials, such as lecithin, by
the maintenance of the required particle size in the case of
dispersions, and by the use of surfactants.
[0135] These compositions may also contain adjuvants such as
preservatives, wetting agents, emulsifying agents and dispersing
agents. Prevention of the action of microorganisms may be ensured
by the inclusion of various antibacterial and antifungal agents,
for example, paraben, chlorobutanol, phenol sorbic acid, and the
like. It may also be desirable to include isotonic agents, such as
sugars, sodium chloride, and the like into the compositions. In
addition, prolonged absorption of the injectable pharmaceutical
form may be brought about by the inclusion of agents that delay
absorption such as aluminum monostearate and gelatin.
[0136] In some cases, in order to prolong the effect of a drug, it
is desirable to slow the absorption of the drug from subcutaneous
or intramuscular injection. This may be accomplished by the use of
a liquid suspension of crystalline or amorphous material having
poor water solubility. The rate of absorption of the drug then
depends upon its rate of dissolution which, in turn, may depend
upon crystal size and crystalline form. Alternatively, delayed
absorption of a parenterally-administered drug form is accomplished
by dissolving or suspending the drug in an oil vehicle.
[0137] Injectable depot forms are made by forming microencapsule
matrices of the subject compounds in biodegradable polymers such as
polylactide-polyglycolide. Depending on the ratio of drug to
polymer, and the nature of the particular polymer employed, the
rate of drug release can be controlled. Examples of other
biodegradable polymers include poly(orthoesters) and
poly(anhydrides). Depot injectable formulations are also prepared
by entrapping the drug in liposomes or microemulsions that are
compatible with body tissue.
[0138] The preparations of the present invention may be given
orally, parenterally, topically, or rectally. They are of course
given by forms suitable for each administration route. For example,
they are administered in tablets or capsule form, by injection,
inhalation, eye lotion, ointment, suppository, etc., administration
by injection, infusion or inhalation; topical by lotion or
ointment; and rectal by suppositories. Oral and/or IV
administration is preferred.
[0139] The phrases "parenteral administration" and "administered
parenterally" as used herein means modes of administration other
than enteral and topical administration, usually by injection, and
includes, without limitation, intravenous, intramuscular,
intraarterial, intrathecal, intracapsular, intraorbital,
intracardiac, intradermal, intraperitoneal, transtracheal,
subcutaneous, subcuticular, intraarticular, subcapsular,
subarachnoid, intraspinal and intrasternal injection and
infusion.
[0140] The phrases "systemic administration," "administered
systemically," "peripheral administration" and "administered
peripherally" as used herein mean the administration of a compound,
drug or other material other than directly into the central nervous
system, such that it enters the patient's system and, thus, is
subject to metabolism and other like processes, for example,
subcutaneous administration.
[0141] These compounds may be administered to humans and other
animals for therapy by any suitable route of administration,
including orally, nasally, as by, for example, a spray, rectally,
intravaginally, parenterally, intracistemally and topically, as by
powders, ointments or drops, including buccally and
sublingually.
[0142] Regardless of the route of administration selected, the
compounds of the present invention, which may be used in a suitable
hydrated form, and/or the pharmaceutical compositions of the
present invention, are formulated into pharmaceutically acceptable
dosage forms by conventional methods known to those of skill in the
art.
[0143] Actual dosage levels of the active ingredients in the
pharmaceutical compositions of this invention may be varied so as
to obtain an amount of the active ingredient which is effective to
achieve the desired therapeutic response for a particular patient,
composition, and mode of administration, without being toxic to the
patient.
[0144] The selected dosage level will depend upon a variety of
factors including the activity of the particular compound of the
present invention employed, or the ester, salt or amide thereof,
the route of administration, the time of administration, the rate
of excretion of the particular compound being employed, the
duration of the treatment, other drugs, compounds and/or materials
used in combination with the particular compound employed, the age,
sex, weight, condition, general health and prior medical history of
the patient being treated, and like factors well known in the
medical arts.
[0145] A physician or veterinarian having ordinary skill in the art
can readily determine and prescribe the effective amount of the
pharmaceutical composition required. For example, the physician or
veterinarian could start doses of the compounds of the invention
employed in the pharmaceutical composition at levels lower than
that required in order to achieve the desired therapeutic effect
and gradually increase the dosage until the desired effect is
achieved.
[0146] In general, a suitable daily dose of a compound of the
invention will be that amount of the compound that is the lowest
dose effective to produce a therapeutic effect. Such an effective
dose will generally depend upon the factors described above.
Generally, intravenous and subcutaneous doses of the compounds of
this invention for a patient, when used for the indicated analgesic
effects, will range from about 0.0001 to about 100 mg per kilogram
of body weight per day, more preferably from about 0.01 to about 50
mg per kg per day, and still more preferably from about 1.0 to
about 100 mg per kg per day. An effective amount is that amount
treats a protein kinase-associated disorder.
[0147] If desired, the effective daily dose of the active compound
may be administered as two, three, four, five, six or more
sub-doses administered separately at appropriate intervals
throughout the day, optionally, in unit dosage forms.
[0148] While it is possible for a compound of the present invention
to be administered alone, it is preferable to administer the
compound as a pharmaceutical composition.
Synthetic Procedure
[0149] Compounds of the present invention are prepared from
commonly available compounds using procedures known to those
skilled in the art, including any one or more of the following
conditions without limitation:
[0150] Within the scope of this text, only a readily removable
group that is not a constituent of the particular desired end
product of the compounds of the present invention is designated a
"protecting group," unless the context indicates otherwise. The
protection of functional groups by such protecting groups, the
protecting groups themselves, and their cleavage reactions are
described for example in standard reference works, such as e.g.,
Science of Synthesis: Houben-Weyl Methods of Molecular
Transformation. Georg Thieme Verlag, Stuttgart, Germany. 2005.
41627 pp. (URL: http://www.science-of-synthesis.com (Electronic
Version, 48 Volumes)); J. F. W. McOmie, "Protective Groups in
Organic Chemistry", Plenum Press, London and New York 1973, in T.
W. Greene and P. G. M. Wuts, "Protective Groups in Organic
Synthesis", Third edition, Wiley, New York 1999, in "The Peptides";
Volume 3 (editors: E. Gross and J. Meienhofer), Academic Press,
London and New York 1981, in "Methoden der organischen Chemie"
(Methods of Organic Chemistry), Houben Weyl, 4th edition, Volume
15/I, Georg Thieme Verlag, Stuttgart 1974, in H.-D. Jakubke and H.
Jeschkeit, "Aminosauren, Peptide, Proteine" (Amino acids, Peptides,
Proteins), Verlag Chemie, Weinheim, Deerfield Beach, and Basel
1982, and in Jochen Lehmann, "Chemie der Kohlenhydrate:
Monosaccharide and Derivate" (Chemistry of Carbohydrates:
Monosaccharides and Derivatives), Georg Thieme Verlag, Stuttgart
1974. A characteristic of protecting groups is that they can be
removed readily (i.e., without the occurrence of undesired
secondary reactions) for example by solvolysis, reduction,
photolysis or alternatively under physiological conditions (e.g.,
by enzymatic cleavage).
[0151] Salts of compounds of the present invention having at least
one salt-forming group may be prepared in a manner known per se.
For example, salts of compounds of the present invention having
acid groups may be formed, for example, by treating the compounds
with metal compounds, such as alkali metal salts of suitable
organic carboxylic acids, e.g., the sodium salt of 2-ethylhexanoic
acid, with organic alkali metal or alkaline earth metal compounds,
such as the corresponding hydroxides, carbonates or hydrogen
carbonates, such as sodium or potassium hydroxide, carbonate or
hydrogen carbonate, with corresponding calcium compounds or with
ammonia or a suitable organic amine, stoichiometric amounts or only
a small excess of the salt-forming agent preferably being used.
Acid addition salts of compounds of the present invention are
obtained in customary manner, e.g., by treating the compounds with
an acid or a suitable anion exchange reagent. Internal salts of
compounds of the present invention containing acid and basic
salt-forming groups, e.g., a free carboxy group and a free amino
group, may be formed, e.g., by the neutralisation of salts, such as
acid addition salts, to the isoelectric point, e.g., with weak
bases, or by treatment with ion exchangers.
[0152] Salts can be converted in customary manner into the free
compounds; metal and ammonium salts can be converted, for example,
by treatment with suitable acids, and acid addition salts, for
example, by treatment with a suitable basic agent.
[0153] Mixtures of isomers obtainable according to the invention
can be separated in a manner known per se into the individual
isomers; diastereoisomers can be separated, for example, by
partitioning between polyphasic solvent mixtures, recrystallisation
and/or chromatographic separation, for example over silica gel or
by, e.g., medium pressure liquid chromatography over a reversed
phase column, and racemates can be separated, for example, by the
formation of salts with optically pure salt-forming reagents and
separation of the mixture of diastereoisomers so obtainable, for
example by means of fractional crystallisation, or by
chromatography over optically active column materials.
[0154] Intermediates and final products can be worked up and/or
purified according to standard methods, e.g., using chromatographic
methods, distribution methods, (re-) crystallization, and the
like.
General Process Conditions
[0155] The following applies in general to all processes mentioned
throughout this disclosure.
[0156] The process steps to synthesize the compounds of the
invention can be carried out under reaction conditions that are
known per se, including those mentioned specifically, in the
absence or, customarily, in the presence of solvents or diluents,
including, for example, solvents or diluents that are inert towards
the reagents used and dissolve them, in the absence or presence of
catalysts, condensation or neutralizing agents, for example ion
exchangers, such as cation exchangers, e.g., in the H.sup.+ form,
depending on the nature of the reaction and/or of the reactants at
reduced, normal or elevated temperature, for example in a
temperature range of from about -100.degree. C. to about
190.degree. C., including, for example, from approximately
-80.degree. C. to approximately 150.degree. C., for example at from
-80 to -60.degree. C., at room temperature, at from -20 to
40.degree. C. or at reflux temperature, under atmospheric pressure
or in a closed vessel, where appropriate under pressure, and/or in
an inert atmosphere, for example under an argon or nitrogen
atmosphere.
[0157] At all stages of the reactions, mixtures of isomers that are
formed can be separated into the individual isomers, for example
diastereoisomers or enantiomers, or into any desired mixtures of
isomers, for example racemates or mixtures of diastereoisomers, for
example analogously to the methods described in Science of
Synthesis: Houben-Weyl Methods of Molecular Transformation. Georg
Thieme Verlag, Stuttgart, Germany. 2005.
[0158] The solvents from which those solvents that are suitable for
any particular reaction may be selected include those mentioned
specifically or, for example, water, esters, such as lower
alkyl-lower alkanoates, for example ethyl acetate, ethers, such as
aliphatic ethers, for example diethyl ether, or cyclic ethers, for
example tetrahydrofuran or dioxane, liquid aromatic hydrocarbons,
such as benzene or toluene, alcohols, such as methanol, ethanol or
f- or 2-propanol, nitriles, such as acetonitrile, halogenated
hydrocarbons, such as methylene chloride or chloroform, acid
amides, such as dimethylformamide or dimethyl acetamide, bases,
such as heterocyclic nitrogen bases, for example pyridine or
N-methylpyrrolidin-2-one, carboxylic acid anhydrides, such as lower
alkanoic acid anhydrides, for example acetic anhydride, cyclic,
linear or branched hydrocarbons, such as cyclohexane, hexane or
isopentane, or mixtures of those solvents, for example aqueous
solutions, unless otherwise indicated in the description of the
processes. Such solvent mixtures may also be used in working up,
for example by chromatography or partitioning.
[0159] The compounds, including their salts, may also be obtained
in the form of hydrates, or their crystals may, for example,
include the solvent used for crystallization. Different crystalline
forms may be present.
[0160] The invention relates also to those forms of the process in
which a compound obtainable as an intermediate at any stage of the
process is used as starting material and the remaining process
steps are carried out, or in which a starting material is formed
under the reaction conditions or is used in the form of a
derivative, for example in a protected form or in the form of a
salt, or a compound obtainable by the process according to the
invention is produced under the process conditions and processed
further in situ.
Prodrugs
[0161] This invention also encompasses pharmaceutical compositions
containing, and methods of treating protein kinase-associated
disorders through administering, pharmaceutically acceptable
prodrugs of compounds of the compounds of the invention. For
example, compounds of the invention having free amino, amido,
hydroxy or carboxylic groups can be converted into prodrugs.
Prodrugs include compounds wherein an amino acid residue, or a
polypeptide chain of two or more (e.g., two, three or four) amino
acid residues is covalently joined through an amide or ester bond
to a free amino, hydroxy or carboxylic acid group of compounds of
the invention. The amino acid residues include but are not limited
to the 20 naturally occurring amino acids commonly designated by
three letter symbols and also includes 4-hydroxyproline,
hydroxylysine, demosine, isodemosine, 3-methylhistidine, norvalin,
beta-alanine, gamma-aminobutyric acid, citrulline homocysteine,
homoserine, ornithine and methionine sulfone. Additional types of
prodrugs are also encompassed. For instance, free carboxyl groups
can be derivatized as amides or alkyl esters. Free hydroxy groups
may be derivatized using groups including but not limited to
hemisuccinates, phosphate esters, dimethylaminoacetates, and
phosphoryloxymethyloxycarbonyls, as outlined in Advanced Drug
Delivery Reviews, 1996, 19, 115. Carbamate prodrugs of hydroxy and
amino groups are also included, as are carbonate prodrugs,
sulfonate esters and sulfate esters of hydroxy groups.
Derivatization of hydroxy groups as (acyloxy)methyl and
(acyloxy)ethyl ethers wherein the acyl group may be an alkyl ester,
optionally substituted with groups including but not limited to
ether, amine and carboxylic acid functionalities, or where the acyl
group is an amino acid ester as described above, are also
encompassed. Prodrugs of this type are described in J. Med. Chem.
1996, 39, 10. Free amines can also be derivatized as amides,
sulfonamides or phosphonamides. All of these prodrug moieties may
incorporate groups including but not limited to ether, amine and
carboxylic acid functionalities.
[0162] Any reference to a compound of the present invention is
therefore to be understood as referring also to the corresponding
pro-drugs of the compound of the present invention, as appropriate
and expedient.
Combinations
[0163] A compound of the present invention may also be used in
combination with other agents, e.g., an additional protein kinase
inhibitor that is or is not a compound of the invention, for
treatment of a protein kinase-associated disorder in a subject.
[0164] By the term "combination" is meant either a fixed
combination in one dosage unit form, or a kit of parts for the
combined administration where a compound of the present invention
and a combination partner may be administered independently at the
same time or separately within time intervals that especially allow
that the combination partners show a cooperative, e.g.,
synergistic, effect, or any combination thereof.
[0165] The compounds of the invention may be administered,
simultaneously or sequentially, with an antiinflammatory,
antiproliferative, chemotherapeutic agent, immunosuppressant,
anti-cancer, cytotoxic agent or kinase inhibitor other than a
compound of the Formula I or salt thereof. Further examples of
agents that may be administered in combination with the compounds
of the invention include, but are not limited to, a PTK inhibitor,
cyclosporin A, CTLA4-Ig, antibodies selected from anti-ICAM-3,
anti-IL-2 receptor, anti-CD45RB, anti-CD2, anti-CD3, anti-CD4,
anti-CD80, anti-CD86, and monoclonal antibody OKT3, agents blocking
the interaction between CD40 and gp39, fusion proteins constructed
from CD40 and gp39, inhibitors of NF-kappa B function,
non-steroidal antiinflammatory drugs, steroids, gold compounds,
antiproliferative agents, FK506, mycophenolate mofetil, cytotoxic
drugs, TNF-.alpha. inhibitors, anti-TNF antibodies or soluble TNF
receptor, rapamycin, leflunimide, cyclooxygenase-2 inhibitors,
paclitaxel, cisplatin, carboplatin, doxorubicin, caminomycin,
daunorubicin, aminopterin, methotrexate, methopterin, mitomycin C,
ecteinascidin 743, porfiromycin, 5-fluorouracil, 6-mercaptopurine,
gemcitabine, cytosine arabinoside, podophyllotoxin, etoposide,
etoposide phosphate, teniposide, melphalan, vinblastine,
vincristine, leurosidine, epothilone, vindesine, leurosine, or
derivatives thereof.
[0166] The compound of the invention and any additional agent may
be formulated in separate dosage forms. Alternatively, to decrease
the number of dosage forms administered to a patient, the compound
of the invention and any additional agent may be formulated
together in any combination. For example, the compound of the
invention inhibitor may be formulated in one dosage form and the
additional agent may be formulated together in another dosage form.
Any separate dosage forms may be administered at the same time or
different times.
[0167] Alternatively, a composition of this invention comprises an
additional agent as described herein. Each component may be present
in individual compositions, combination compositions, or in a
single composition.
EXEMPLIFICATION OF THE INVENTION
[0168] The invention is further illustrated by the following
examples, which should not be construed as further limiting. The
practice of the present invention will employ, unless otherwise
indicated, conventional techniques of cell biology, cell culture,
molecular biology, transgenic biology, microbiology and immunology,
which are within the skill of the art.
General Synthesis Methods
[0169] All starting materials, building blocks, reagents, acids,
bases, dehydrating agents, solvents, and catalysts utilized to
synthesis the compounds of the present invention are either
commercially available or can be produced by organic synthesis
methods known to one of ordinary skill in the art (Houben-Weyl 4th
Ed. 1952, Methods of Organic Synthesis, Thieme, Volume 21).
Further, the compounds of the present invention can be produced by
organic synthesis methods known to one of ordinary skill in the art
as shown in the following examples.
LIST OF ABBREVIATIONS
[0170] BINAP (.+-.)-(1,1'-binaphthalene-2-2'
diyl)bis(diphenylphosphine)
DIEA Diethylamine
DIPEA Diisoproylethylamine
DMF Dimethylformamide
[0171] HPLC High pressure liquid chromatography HRMS High
resolution mass spectrometry HBTU
O-Benzotriazol-1-yl-N,N,N',N'-tetramethyluronium
hexafluorophosphate
HOBt 1-Hydroxy-1H-benzotriazol
[0172] LC/MS Liquid chromatography/mass spectrometry
NMM N-methylmorpholine
NMP N-methylpyrrolidine
[0173] RT room temperature
THF Tetrahydrofuran
Et Ethyl
NBS N-Bromosuccinimide
[0174] DIAD Diisopropyl azo dicarboxylate
Ts Tosyl
[0175] TBAF Tetra-n-butylammonium fluoride
Example 1
(5-Bromo-2-chloro-pyrimidin-4-yl)-(1-ethyl-propyl)-amine
##STR00181##
[0177] To a solution of 5-Bromo-2,4-dichloropyrimidine (4.56 g, 20
mmol) in Ethanol (9 mL) is added 1-Ethylpropylamine (2.6 mL, 22
mmol) and DIEA (7 mL, 40 mmol) at ambient temperature. The reaction
mixture is stirred at ambient temperature for 16 hrs, then is
concentrated in vacuo and the residue is purified by flash
chromatography (silica gel, ethyl acetate:hexane=3:97 to 30:70) to
give (5-Bromo-2-chloro-pyrimidin-4-yl)-(1-ethyl-propyl)-amine. MS
(ESI)m/z 280 (M+H).sup.+. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta.
8.1 (s, 1H), 5.24 (d, 1H), 4.1 (m, 1H), 1.58 (m, 4H), 0.93 (t,
6H).
Example 2
Tributyl-((Z)-2-ethoxy-vinyl)-stannane
##STR00182##
[0179] To a solution of Ethyl ethynyl ether (2.26 mL, 50% in
hexane, 15 mmol) in toluene (40 mL) is added Tri-n-butyl hydride
(2.7 mL, 10 mmol) and AIBN (81 mg, 0.5 mmol) at ambient
temperature. The reaction mixture is heated at 100.degree. C. for
16 hrs. After cooling down, the mixture is concentrated in vacuo to
give tributyl-((Z)-2-ethoxy-vinyl)-stannane. The crude product is
used as is.
Example 3
[2-Chloro-5-((Z)-2-ethoxy-vinyl)-pyrimidin-4-yl]-(1-ethyl-propyl)-amine
##STR00183##
[0181] To a solution of crude compound from example 2 (4.25 g,
.about.75%, 8.8 mmol) in CH-3CN (10 mL) is added
(5-Bromo-2-chloro-pyrimidin-4-yl)-(1-ethyl-propyl)-amine (2.25 g, 8
mmol), Et.sub.4NCl (1.33 g, 8 mmol) and Pd(PPh.sub.3).sub.2Cl.sub.2
(280 mg, 0.4 mmol) at ambient temperature. The reaction mixture is
purged with N.sub.2, sealed in a microwave reactor and heated at
100.degree. C. for 17 mins. After cooling down the mixture is
concentrated in vacuo and the residue is purified by flash
chromatography (silica gel, ethyl acetate:hexane=5:95 to 40:60) to
give
[2-Chloro-5-((Z)-2-ethoxy-vinyl)-pyrimidin-4-yl]-(1-ethyl-propyl)-amine.
MS (ESI)m/z 270 (M+H).sup.+. .sup.1H NMR (CDCl.sub.3, 400 MHz).
.delta. 8.02 (s, 1H), 6.26 (d, 1H), 5.46 (d, 1H), 4.91 (d, 1H),
4.16 (m, 1H), 3.99 (q, 2H), 1.60-1.69 (m, 2H), 1.43-1.52 (m, 2H),
1.32 (t, 3H), 0.92 (t, 6H).
Example 7
2-Chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine
##STR00184##
[0183] To a solution of
[2-Chloro-5-((Z)-2-ethoxy-vinyl)-pyrimidin-4-yl]-(1-ethylpropyl)-amine
(1.1 g, 4.07 mmol) in EtOH (8 mL) is added concentrated HCl (0.1
mL) at ambient temperature. The reaction mixture is sealed in a
microwave reactor and heated at 100.degree. C. for 10 mins. After
cooling down the mixture is concentrated in vacuo to provide
2-Chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine. The crude
product is used as it is. The material can be purified by flash
chromatography (SiO2, EtOAc:Hexane=1:5).
[0184] MS (ESI)m/z 224 (M+H).sup.+. .sup.1H NMR (CDCl.sub.3, 400
MHz). .delta. 8.87 (s, 1H), 7.30 (d, 1H), 6.69 (d, 1H), 4.69 (m,
1H), 1.77-1.99 (m, 4H), 0.77 (t, 6H)
Example 8
5,5-Dibromo-2-chloro-7-(1-ethyl-propyl)-5,7-dihydro-pyrrolo[2,3-d]pyrimidi-
n-6-one
##STR00185##
[0186] To a mixture of
2-Chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine (crude,
4.07 mmol) in t-BuOH (7 mL) is added 2 mL of H.sub.2O at ambient
temperature, then NBS (2.28 g, 12.8 mmol) is added to the orange
color solution. The mixture is stirred at 28-30.degree. C. for 2.5
hrs, then is concentrated and taken up in ethyl acetate, washed
with NaHCO.sub.3 aqueous solution, and brine. The organics are
dried with Na.sub.2SO.sub.4, filtered and concentrated to provide
5,5-Dibromo-2-chloro-7-(1-ethyl-propyl)-5,7-dihydro-pyrrolo[2,3-d]pyrimid-
in-6-one. The crude product is used as it is.
[0187] MS (ESI)m/z 398 (M+H).sup.+.
Example 9
2-Chloro-7-(1-ethyl-propyl)-5,7-dihydro-pyrrolo[2,3-d]pyrimidin-6-one
##STR00186##
[0189] To a solution of
5,5-Dibromo-2-chloro-7-(1-ethyl-propyl)-5,7-dihydro-pyrrolo[2,3-d]pyrimid-
in-6-one (crude, .about.5.3 mmol) in acetic acid (6 mL) and THF (4
mL) is added Zn dust (1.37 g, 21 mmol) at 0.degree. C. The mixture
is stirred at 0.degree. C. for 2 mins then heated to room
temperature, stirring for 30 mins. The mixture is filtered through
celite, rinsed with ethyl acetate. The filtrate is concentrated in
vacuo and the residue is purified by flash chromatography (ethyl
acetate:hexane=5:95 to 40:60) to give
2-Chloro-7-(1-ethyl-propyl)-5,7-dihydro-pyrrolo[2,3-d]pyrimidin-6-one.
[0190] MS (ESI)m/z 240 (M+H).sup.+. .sup.1H NMR (CDCl.sub.3, 400
MHz). .delta. 8.17 (s, 1H), 4.20 (m, 1H), 3.58 (s, 2H), 2.10 (m,
2H), 1.84 (m, 2H), 0.84 (t, 6H).
Examples 10-13
[0191] By repeating the procedures described in example 6-9, using
appropriate starting materials, the following compounds are
obtained.
TABLE-US-00005 Structure MS (m/z) (M + 1) ##STR00187## 210
##STR00188## 252 ##STR00189## 238 ##STR00190## 288
Example 14
(3-Amino-phenyl)-(4-methyl-piperazin-1-yl)-methanone
##STR00191##
[0193] A solution of 3-aminobenzoic acid (1.51 g, 11 mmol),
1-methylpiperazine (1.1 mL, 10 mmol), EDCI HCl (2.87 g, 15 mmol)
and Et.sub.3N (2.8 mL, 20 mmol) in CH.sub.2Cl.sub.2(10 mL) is
stirred at room temperature for 20 hours. Then saturated
NaHCO.sub.3 aqueous solution is added. The aqueous layer was
extracted with CH.sub.2Cl.sub.2, and the organic extracts were
dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure. The crude product is purified by column
chromatography (SiO.sub.2, MeOH: CH.sub.2Cl.sub.2=0.7:99.3 to 6:93)
to give 1.75 g of the title compound as a yellow solid.
[0194] MS (ESI)m/z 220 (M+H).sup.+
Examples 15-20
[0195] By repeating the procedures described in example 14, using
appropriate starting materials, the following compounds are
obtained.
TABLE-US-00006 Structure MS (m/z) (M + 1) ##STR00192## 237
##STR00193## 250 ##STR00194## 225 ##STR00195## 238 ##STR00196## 207
##STR00197## 220
Example 21
N-(4-Methoxy-3-nitro-phenyl)-isonicotinamide
##STR00198##
[0197] A mixture of 4-methoxy-3-nitroaniline (168 mg, 1 mmol) and
Isonicotinoyl chloride hydrogen chloride (267 mg, 0.2 M in 1.5
mmol) in pyridine (1 mL) is sealed in a microwave reactor and
heated at 100.degree. C. under microwave radiation for 5 mins. Then
1N NaOH aqueous solution is added to the reaction mixture. After
stirring at room temperature for several minutes, the mixture is
filtered. The solid is washed with H.sub.2O and air dried to give
263 mg of the title compound as a yellow solid.
[0198] MS (ESI)m/z 274 (M+H).sup.+
Example 22
N-(4-Floro-3-nitro-phenyl)-isonicotinamide
##STR00199##
[0200] The same procedure is repeated as described in example 21 to
give the title compound as a pink solid.
[0201] MS (ESI)m/z 262 (M+H).sup.+.
Example 23
N-(3-Amino-4-floro-phenyl)-isonicotinamide
##STR00200##
[0203] A mixture of 4-floro-3-nitroaniline (100 mg, 0.38 mmol) and
Tin chloride (180 mg, 0.95 mmol) in EtOH (1 mL) with 4 drops of
concentrated HCl is heated at 80.degree. C. for 4 hour. Then
saturated NaHCO.sub.3 aqueous solution is added. The aqueous layer
was extracted with EtOAc, and the organic extracts are washed with
brine, dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure. The crude product is purified by column
chromatography (SiO2, MeOH:CH.sub.2Cl.sub.2=1:99 to 10:90) to give
55.5 mg of the title compound as yellow solid.
[0204] MS (ESI)m/z 232(M+H).sup.+
Example 24
N-(3-Amino-4-methoxy-phenyl)-isonicotinamide
##STR00201##
[0206] The same procedure can be repeated as described in example
23 to give the title compound as a pink solid.
[0207] MS (ESI)m/z 244 (M+H).sup.+.
Example 25
1-(4-Nitro-phenyl)-piperidin-4-one
##STR00202##
[0209] To a solution of 1-(4-Nitro-phenyl)-piperidin-4-ol (100 mg,
0.45 mmol) in CH.sub.2Cl.sub.2(2 mL) is added Dess-Martin
periodinane (286 mg, 0.675 mmol) at for 2.5 hours. The reaction is
quenched with 1N NaOH aqueous solution. The aqueous layer is
extracted with CH.sub.2Cl.sub.2, and the organic extracts are dried
over Na.sub.2SO.sub.4, filtered and concentrated under reduced
pressure. The crude product is purified by column chromatography
(SiO2, EtOAc:Hexane=12:88 to 100:0) to give 84 mg of the title
compound as a white solid.
[0210] MS (ESI)m/z 221 (M+H).sup.+
Example 26
1-Methyl-4-[1-(4-nitro-phenyl)-piperidin-4-yl]piperazine
##STR00203##
[0212] A mixture of 1-(4-Nitro-phenyl)-piperidin-4-one (84 mg, 0.38
mmol) and 1-methylpiperazine (0.085 mL, 0.76 mmol) in MeOH (2 mL)
is stirred at room temperature for 5 hours. Then to the reaction
mixture is added 0.2 mL of HOAc, followed by NaCNBH.sub.3 (72 mg,
1.14 mmol). The mixture is stirred at room temperature for 0.5
hour, then concentrated. The residue is taken up in EtOAc, washed
with saturated NaHCO.sub.3 aqueous solution and brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The crude product is purified by column chromatography (SiO2, 2N
NH.sub.3 in MeOH:CH.sub.2Cl.sub.2=1:99 to 10:90) to give 46 mg of
the title compound as a yellow solid.
[0213] MS (ESI)m/z 305 (M+H).sup.+
Example 27
4-[4-(4-Methyl-piperazin-1-yl)-piperidin-1-yl]-phenylamine
##STR00204##
[0215] A suspension of
1-Methyl-4-[1-(4-nitro-phenyl)-piperidin-4-yl]-piperazine (46 mg,
0.15 mmol) and Pd/C (10%, 8 mg) in MeOH (2 mL) is stirred at room
temperature under H-.sub.2(balloon pressure) for 16 hours, then
filtered through celite, washed with EtOAc, concentrated under
reduced pressure to give 42 mg of the title compound as a light
grey solid.
[0216] MS (ESI)m/z 275 (M+H).sup.+
Example 28
Benzoic acid 1-(4-amino-phenyl)-piperidin-4-yl ester
##STR00205##
[0218] The same procedure is repeated as described in example 23 to
give the title compound as a pink solid.
[0219] MS (ESI)m/z 297(M+H).sup.+.
Example 29
3-(2-Pyrrolidin-1-yl-ethoxy)-phenylamine
##STR00206##
[0221] To a mixture of PPh.sub.3 (866 mg, 3.3 mmol) in THF (6 mL)
is added DIAD (0.65 mL, 3.3 mmol) at 0.degree. C. The suspension
was stirred for 10 minutes, then heated to room temperature. To the
mixture, 4-nitrophenol (460 mg, 3.3 mmol) and
1-(2-hydroxyethyl)-pyrrolidine (0.26 mL, 2.2 mmol) is added, and
the mixture is stirred at room temperature for 16 hours, then
concentrated. The residue is taken up in EtOAc, washed with 1N NaOH
aqueous solution and brine, dried over Na.sub.2SO.sub.4, filtered
and concentrated under reduced pressure. The crude product is
purified by column chromatography (SiO2, MeOH:CH.sub.2Cl.sub.2=1:99
to 10:90) to give 277 mg of
1-[2-(4-Nitro-phenoxy)-ethyl]-pyrrolidine as a white solid.
[0222] MS (ESI)m/z 237 (M+H).sup.+
[0223] The same procedure is repeated as described in example 27 by
using 1-[2-(4-Nitro-phenoxy)-ethyl]-pyrrolidine as a starting
material to give the title compound as a yellow oil.
[0224] MS (ESI)m/z 207(M+H).sup.+.
Example 30-33
[0225] By repeating the procedures described in example 29, using
appropriate starting materials, the following compounds are
obtained.
TABLE-US-00007 Structure MS (m/z) (M + 1) ##STR00207## 207
##STR00208## 223 ##STR00209## 236 ##STR00210## 209
Example 34
(3-Nitro-phenyl)-(2-pyrrolidin-1-yl-ethyl)-amine
##STR00211##
[0227] The mixture of 1-fluoro-3-nitrobenzene (420 mg, 3 mmol) in
DMF (1.5 mL), N-(2-aminoethyl)-pyrrolidine (514 mg, 4.5 mmol) and
Cs.sub.2CO.sub.3(977 mg, 3 mmol) is heated at 1-00.degree. C. under
microwave radiation for 2.5 hours, then concentrated. The mixture
is diluted with EtOAc, washed with saturated NaHCO.sub.3 aqueous
solution and brine, dried over Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. The crude product is purified
by column chromatography (SiO2, MeOH:CH.sub.2Cl.sub.2=1:99 to
10:90) to give 130 mg of the title compound as a light brown
oil.
[0228] MS (ESI)m/z 236 (M+H).sup.+
Example 35
[2-(4-Methyl-piperazin-1-yl)-ethyl]-(3-nitro-phenyl)-amine
##STR00212##
[0230] The same procedure is repeated as described in example 34 to
give the title compound as a yellow oil.
[0231] MS (ESI)m/z 265(M+H).sup.+.
Example 36
N-(2-Pyrrolidin-1-yl-ethyl)-benzene-1,3-diamine
##STR00213##
[0233] The same procedure is repeated as described in example 27 to
give the title compound as a light brown oil.
[0234] MS (ESI)m/z 206(M+H).sup.+.
Example 37
N-[2-(4-Methyl-piperazin-1-yl)-ethyl]-benzene-1,3-diamine
##STR00214##
[0236] The same procedure is repeated as described in example 27 to
give the title compound as a light brown oil.
[0237] MS (ESI)m/z 235(M+H).sup.+.
Example 38
1-Methyl-4-(6-nitro-pyridin-3-yl)-piperazine
##STR00215##
[0239] A mixture of 5-bromo-2-nitropyridine (500 mg, 2.46 mmol) and
1-methylpiperazine (1 mL) is heated at 80.degree. C. for 2 hour.
Then water is added. The aqueous layer is extracted with EtOAc, and
the organic extracts were washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The crude product is purified by column chromatography (SiO2,
MeOH:CH.sub.2Cl.sub.2=0.7:99.3 to 6:93) to give 520 mg of the title
compound as yellow solid.
[0240] MS (ESI)m/z 223 (M+H).sup.+
Example 39
1-[4-(6-Nitro-pyridin-3-yl)-piperazin-1-yl]-ethanone
##STR00216##
[0242] To a mixture of 5-bromo-2-nitropyridine (406 mg, 2 mmol) and
1-acetylpiperazine (256 mg, 2 mmol) in toluene (5 mL) is added
Cs.sub.2CO.sub.3, then Pd2(dba)3(74 mg, 0.08 mmol) and BINAP (100
mg, 0.16 mmol) are added. The mixture is degassed, and heated at
100.degree. C. for 16 hours. Then the mixture is cooled down to
room temperature, diluted with EtOAc, and filtered through celite.
The filtrate is concentrated under reduced pressure. The crude
product is purified by column chromatography (SiO2,
MeOH:CH.sub.2Cl.sub.2=0.7:99.3 to 6:93) to give 270 mg of the title
compound as yellow solid.
[0243] MS (ESI)m/z 251 (M+H).sup.+
Example 40
5-(4-Methyl-piperazin-1-yl)-pyridin-2-ylamine
##STR00217##
[0245] The same procedure is repeated as described in example 27 to
give the title compound as a light brown solid.
[0246] MS (ESI)m/z 193(M+H).sup.+.
Example 41
1-[4-(6-Amino-pyridin-3-yl)-piperazin-1-yl]-ethanone
##STR00218##
[0248] The same procedure is repeated as described in example 27 to
give the title compound as a brown solid.
[0249] MS (ESI)m/z 221(M+H).sup.+.
Example 42
1-[4-(4-Nitro-phenyl)-piperidin-1-yl]-ethanone
##STR00219##
[0251] To a solution of 4-(4-Nitro-phenyl)-piperidin (206 mg, 1
mmol) in CH.sub.2Cl.sub.2(3 mL) is added AcCl (0.106 mL, 1.5 mmol)
at 0.degree. C. Then Et.sub.3N (0.253 mL, 1.8 mmol) is added
slowly. The mixture is stirred at 0.degree. C. for 10 minutes. Then
saturated NaHCO.sub.3 aqueous solution is added. The aqueous layer
is extracted with CH.sub.2Cl.sub.2, and the organic extracts are
dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure. The crude product is purified by column
chromatography (SiO.sub.2, MeOH:CH.sub.2Cl.sub.2=0.7:99.3 to 6:93)
to give 273 mg of the title compound as yellow solid.
[0252] MS (ESI)m/z 249 (M+H).sup.+
Example 43
1-[4-(4-Amino-phenyl)-piperidin-1-yl]-ethanone
##STR00220##
[0254] The same procedure is repeated as described in example 27 to
give the title compound as a yellow solid.
[0255] MS (ESI)m/z 219 (M+H).sup.+.
Example 44
7-(1-Ethyl-propyl)-2-[3-fluoro-4-(4-methyl-piperazin-1-yl)-phenylamino]-5,-
7-dihydro-pyrrolo[2,3-d]pyrimidin-6-one
##STR00221##
[0257] To a mixture of
2-Chloro-7-(1-ethyl-propyl)-5,7-dihydro-pyrrolo[2,3-d]pyrimidin-6-one
(18 mg, 0.075 mmol) and TsOH (1.12 ml, 0.2 M in 1,4 dioxane) is
added 3-Fluoro-4-(4-methylpiperazinn)aniline (23.5 mg, 0.1125
mmol), and DMF (0.25 mL) at ambient temperature. The reaction
mixture is sealed in a microwave reactor and heated at 140.degree.
C. for 30 mins. The mixture is diluted with EtOAc, washed with
NaHCO.sub.3 aqueous solution and brine, dried (Na.sub.2SO.sub.4),
filtered and concentrated. The crude product is purified by
prep-HPLC to give 27 mg of the title compound as a brown solid.
[0258] MS (ESI)m/z 413 (M+H).sup.+.
[0259] .sup.1H NMR (DMSO, 400 MHz). .delta. 9.48 (s, 1H), 8.09 (s,
1H), 7.72 (d, 1H), 7.34 (d, 1H), 6.96 (t, 1H), 4.08 (m, 1H), 3.60
(s, 2H), 2.96 (s, 4H), 2.51 (s, 2H), 2.10 (m, 2H), 1.78 (m, 2H),
0.79 (t, 6H).
Examples 45-90
[0260] By repeating the procedures described in example 44, using
appropriate starting materials, the following compounds are
obtained.
TABLE-US-00008 ##STR00222## MS Found Ar (M + H) ##STR00223## 395
##STR00224## 423 ##STR00225## 382 ##STR00226## 380 ##STR00227## 423
##STR00228## 453 ##STR00229## 441 ##STR00230## 440 ##STR00231## 410
##STR00232## 428 ##STR00233## 410 ##STR00234## 423 ##STR00235## 396
##STR00236## 354 ##STR00237## 447 ##STR00238## 435 ##STR00239## 417
##STR00240## 466 ##STR00241## 459 ##STR00242## 459 ##STR00243## 382
##STR00244## 364 ##STR00245## 378 ##STR00246## 337 ##STR00247## 500
##STR00248## 396 ##STR00249## 478 ##STR00250## 410 ##STR00251## 410
##STR00252## 407 ##STR00253## 426 ##STR00254## 439 ##STR00255## 412
##STR00256## 409 ##STR00257## 438 ##STR00258## 354 ##STR00259## 368
##STR00260## 382 ##STR00261## 408 ##STR00262## 368 ##STR00263## 396
##STR00264## 436 ##STR00265## 466 ##STR00266## 451 ##STR00267## 396
##STR00268## 424
Examples 91-93
[0261] By repeating the procedures described in example 44, using
appropriate starting materials, the following compounds are
obtained.
TABLE-US-00009 ##STR00269## MS Found Ar (M + H) ##STR00270## 455
##STR00271## 458 ##STR00272## 407
Examples 94-97
[0262] By repeating the procedures described in example 44, using
appropriate starting materials, the following compounds are
obtained.
TABLE-US-00010 ##STR00273## MS Found Ar (M + H) ##STR00274## 380
##STR00275## 352 ##STR00276## 365 ##STR00277## 393
Example 98-99
[0263] By repeating the procedures described in example 44, using
appropriate starting materials, the following compounds are
obtained.
TABLE-US-00011 ##STR00278## MS Found Ar (M + H) ##STR00279## 380
##STR00280## 393
Examples 100-101
[0264] By repeating the procedures described in example 44, using
appropriate starting materials, the following compounds are
obtained.
TABLE-US-00012 ##STR00281## MS Found Ar (M + H) ##STR00282## 407
##STR00283## 435
Example 102
1-(1-{4-[7-(1-Ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino]-phenyl}--
piperidin-4-yl)-ethanone
##STR00284##
[0266] To a mixture of
1-[4-(4-Amino-phenyl)-piperazin-1-yl]-ethanone (70.5 mg, 0.32 mmol)
and NaOtBu (38.4 mg, 0.4 mmol) in 1,4-dioxane (0.3 mL) is added a
solution of 2-Chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine
(60 mg, 0.26 mmol) in 1,4-dioxane (0.6 mL) and a suspension of
Pd.sub.2(dba).sub.3(12.2 mg, 0.013 mmol) and BINAP (16.6 mg, 0.026
mmol). The mixture is degassed, and heated at 100.degree. C. for 3
hours. Then the mixture is cooled down to room temperature, diluted
with EtOAc, and filtered through celite. The filtrate is
concentrated under reduced pressure. The crude product is purified
prep-HPLC to give 84.9 mg of the title compound as pale white
solid.
[0267] MS (ESI)m/z 407 (M+H).sup.+
Example 103-117
[0268] By repeating the procedures described in example 102, using
appropriate starting materials, the following compounds are
obtained.
TABLE-US-00013 ##STR00285## MS Found Ar (M + H) ##STR00286## 395
##STR00287## 417 ##STR00288## 406 ##STR00289## 431 ##STR00290## 364
##STR00291## 406 ##STR00292## 397 ##STR00293## 380 ##STR00294## 408
##STR00295## 394 ##STR00296## 392 ##STR00297## 394 ##STR00298## 391
##STR00299## 273 ##STR00300## 245
Example 118
(2-Chloro-5-nitro-pyrimidin-4-yl)-(1-ethyl-propyl)-amine
##STR00301##
[0270] To a solution of 2,4-dichloro-5-nitro-pyrimidine (2 g, 10.31
mmol) in anhydrous EtOH (20 ml) is added 1-ethylpropylamine (1.322
ml, 11.341 mmol) at 0 C (ice bath) under inert atmosphere. To this
is added neat DIPEA (2.694 ml, 15.465 mmol). The reaction is
stirred at r.t. for 8 hrs. The reaction mixture was concentrated in
vacuo and the residue is dissolved with EtOAc. The organic layer is
washed with sat. NaHCO.sub.3 and brine, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. Purification with
column chromatography (SiO.sub.2, 1:3 EtOAC/Hexane) gives the
desired product.
[0271] MS (ESI)m/z 245.1
Example 119
(2-Chloro-5-amino-pyrimidin-4-yl)-(1-ethyl-propyl)-amine
##STR00302##
[0273] To a solution of
(2-Chloro-5-nitro-pyrimidin-4-yl)-(1-ethyl-propyl)-amine (1 g,
4.087 mmol) in anhydrous EtOH (50 ml) is added Tin(II) chloride
(2.324 g, 12.2607 mmol) and concentrated HCl (1 ml) at ambient
temperature. The reaction is heated to 80.degree. C. for 1 h and
quenched with 1N NaOH at 0.degree. C. The mixture is extracted with
EtOAc, washed with brine, dried over Na2SO4, and concentrated in
vacuo to give the crude product. The crude is used as is.
[0274] MS (ESI)m/z 215.2
Example 120
2-Chloro-9-(1-ethyl-propyl)-7,9-dihydro-purin-8-one
##STR00303##
[0276] To a microwave vial is added the crude
(2-Chloro-5-amino-pyrimidin-4-yl)-(1-ethyl-propyl)-amine (0.5 g,
2.329 mmol) and anhydrous DMF (15 ml) followed by
1,1'-carbonyldiimidazole (1.133 g, 6.987 mmol). Sealed vial and
microwave heated at 100.degree. C. for 10 min. The reaction mixture
is diluted with EtOAc, washed with water, dried over Na2SO4, and
concentrated in vacuo. Purification with column chromatography
(SiO2, 1:1 EtOAC/Hexane) gives the desired product.
[0277] MS (ESI) m/z 241.1
Example 121
2-[4-(4-Acetyl-piperazin-1-yl)-phenylamino]-9-(1-ethyl-propyl)-7,9-dihydro-
-purin-8-one
##STR00304##
[0279] By repeating the procedures described in example 44, using
2-Chloro-9-(1-ethyl-propyl)-7,9-dihydro-purin-8-one as a starting
material, the desired product is obtained.
[0280] MS (ESI) 424.2
Example 122
2-Chloro-9-(1-ethyl-propyl)-7-methyl-7,9-dihydro-purin-8-one
##STR00305##
[0282] To a solution of
2-Chloro-9-(1-ethyl-propyl)-7,9-dihydro-purin-8-one (100 mg, 0.41
mmol) in anhydrous DMF (2 ml) is added methyl iodide (21 ul, 0.41
mmol) followed by NaH (50%, 22 mg, 0.4571 mmol). The reaction is
stirred under nitrogen for 1.5 h. The reaction mixture is quenched
with ice water and extracted with EtOAc. The extracts are dried
over Na.sub.2SO.sub.4 and concentrated in vacuo to give the crude
2-Chloro-9-(1-ethyl-propyl)-7-methyl-7,9-dihydro-purin-8-one. The
crude product is used as is.
[0283] MS (ESI)m/z 255.1
Example 123
2-[4-(4-Acetyl-piperazin-1-yl)-phenylamino]-9-(1-ethyl-propyl)-7-methyl-7,-
9-dihydro-purin-8-one
##STR00306##
[0285] By repeating the procedures described in example 44, using
2-Chloro-9-(1-ethyl-propyl)-7-methyl-7,9-dihydro-purin-8-one as a
starting material, the desired product is obtained.
[0286] MS (ESI)m/z 438.2
Example 124
Allyl-(1-ethyl-propyl)-amine
##STR00307##
[0288] To a solution of 3-pentanone (1 g, 11.61 mmol) in anhydrous
1,2-dichloroethane (45 ml) is added allylamine (0.872 ml, 11.61
mmol) followed by NaBH(OAc).sub.3 (3.44 g, 16.254 mmol) at ambient
temperature and under nitrogen. The reaction is stirred at room
temperature overnight. The reaction mixture is quenched with 1N
NaOH and extracted with dichloromethane. The extract is dried over
Na.sub.2SO.sub.4 and concentrated in vacuo to
allyl-(1-ethyl-propyl)-amine
Example 125
Allyl-(5-bromo-2-chloro-pyrimidin-4-yl)-(1-ethyl-propyl)-amine
##STR00308##
[0290] To a solution of allyl-(1-ethyl-propyl)-amine (10 mmol) is
added anhydrous isopropanol (50 ml) and
5-bromo-2,4-dichloropyrimidine (2.979 g, 5 mmol) followed by
diisopropylethylamine (2.61 ml, 15 mmol) at ambient temperature.
The reaction is stirred overnight and concentrated in vacuo. The
residue is purified with column chromatography (SiO2, 1:5
EtOAC/Hexane) to give the desired product.
[0291] MS (ESI)m/z 320.0
Example 126
2-Chloro-7-(1-ethyl-propyl)-5-methyl-7H-pyrrolo[2,3-d]pyrimidine
##STR00309##
[0293] To a solution of
allyl-(5-bromo-2-chloro-pyrimidin-4-yl)-(1-ethyl-propyl)-amine
(2.76 g, 8.7 mmol) in anhydrous DMF (15 ml) was added 8 mol % of
Pd(OAc).sub.2 (156 mg, 0.69 mmol) and 8 mol % of PPh3 (182 mg, 0.69
mmol) and triethylamine (2.4 ml, 17.3 mmol) a The reaction is
stirred at 100.degree. C. overnight. The reaction mixture is
diluted with EtOAc, washed with water, dried over Na2SO4, and
concentrated in vacuo. Purification with column chromatography
(SiO2, 1:2 EtOAC/Hexane) gives the desired product.
[0294] MS (ESI) m/z 238.2
Example 127
1-(4-{4-[7-(1-Ethyl-propyl)-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino]-
-phenyl}-piperazin-1-yl)-ethanone
##STR00310##
[0296] By repeating the procedures described in example 102, using
2-Chloro-7-(1-ethyl-propyl)-5-methyl-7H-pyrrolo[2,3-d]pyrimidine as
a starting material, the desired product is obtained.
[0297] MS (ESI)m/z 421.2
Example 128
(2-Chloro-5-prop-1-ynyl-pyrimidin-4-yl)-(1-ethyl-propyl)-amine
##STR00311##
[0299] To a microwave vial is added a solution of
(5-Bromo-2-chloro-pyrimidin-4-yl)-(1-ethyl-propyl)-amine (0.5 g,
1.80 mmol) in anhydrous toluene (10 ml), tributyl(1-propynyl)-tin
(1.1 ml, 3.6 mmol) and 2 mol % of Pd(PPh3).sub.4 (41.5 mg, 0.036
mmol). The reaction is heated at 120.degree. C. for 1 hr by
employing microwave. The reaction mixture is diluted with EtOAc,
washed with sat. NaHCO.sub.3 aqueous solution and water, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. Purification with
column chromatography (SiO2, 1:5 EtOAC/Hexane) gives 0.32 g of the
desired product.
[0300] MS (ESI)m/z 238.2
Example 129
2-Chloro-7-(1-ethyl-propyl)-6-methyl-7H-pyrrolo[2,3-d]pyrimidine
##STR00312##
[0302] In a microwave vial is added
(2-Chloro-5-prop-1-ynyl-pyrimidin-4-yl)-(1-ethyl-propyl)-amine
(0.22 g, 0.92 mmol), anhydrous DMF (3 ml), and CuI (53 mg, 0.27
mmol). The reaction is heated at 160.degree. C. for 1 hr by
employing microwave. The reaction mixture is diluted with EtOAc,
washed with sat. NaHCO3 aqueous solution and water, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. Purification with
column chromatography (SiO.sub.2, 1:4 EtOAC/Hexane) gives 43 mg of
the desired product.
[0303] MS (ESI)m/z 238.2.
Example 130
1-(4-{4-[7-(1-Ethyl-propyl)-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino]-
-phenyl}-piperazin-1-yl)-ethanone
##STR00313##
[0305] By repeating the procedures described in example 102, using
2-Chloro-7-(1-ethyl-propyl)-6-methyl-7H-pyrrolo[2,3-d]pyrimidine as
a starting material, the desired product is obtained.
[0306] MS (ESI)m/z 421.4
Example 131
[7-(1-Ethyl-propyl)-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-(4-piperazin-
-1-yl-phenyl)-amine
##STR00314##
[0308] By repeating the procedures described in example 102, using
2-Chloro-7-(1-ethyl-propyl)-6-methyl-7H-pyrrolo[2,3-d]pyrimidine as
a starting material, the desired product is obtained.
[0309] MS (ESI)m/z 379.1
Example 132
[2-Chloro-5-(3,3-diethoxy-prop-1-ynyl)-pyrimidin-4-yl]-(1-ethyl-propyl)-am-
ine
##STR00315##
[0311] To a mixture of
(5-Bromo-2-chloro-pyrimidin-4-yl)-(1-ethyl-propyl)-amine (420 mg,
1.5 mmol) and propiolaldehyde diethyl acetal (0.32 mL, 2.25 mmol)
in DMF (6 mL) is added PdCl.sub.2(PPh.sub.3).sub.2(105 mg, 0.15
mmol) and CuI (28 mg, 0.15 mmol), followed by Et.sub.3N (0.42 mL, 3
mmol). The mixture is degassed and heated at 55.degree. C. for 16
h. Then the mixture is cooled down to room temperature, diluted
with EtOAc, washed with water and brine. The organic layer is dried
(Na.sub.2SO.sub.4), filtered and concentrated under reduced
pressure. The crude product is purified by column chromatography
(SiO2, EtOAc:Heptane=5:95 to 40:60) to give 182 mg of the title
compound as a light brown oil.
[0312] MS (ESI)m/z 326 (M+H).sup.+
Example 133
2-Chloro-6-diethoxymethyl-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine
##STR00316##
[0314] To a solution of
[2-Chloro-5-(3,3-diethoxy-prop-1-ynyl)-pyrimidin-4-yl]-(1-ethyl-propyl)-a-
mine (326 mg, 1 mmol) in THF (2 mL) is added a solution of 1M TBAF
in THF (5 mL, 5 mmol) at ambient temperature. The reaction mixture
is heated at 68.degree. C. for 2 hours. After cooling down, the
mixture is concentrated in vacuo. The crude product is purified by
column chromatography (SiO2, EtOAc:Heptane=5:95 to 40:60) to give
307 mg of the title compound as a colorless oil.
[0315] MS (ESI)m/z 326 (M+H).sup.+.
Example 134
1-(4-{4-[6-Diethoxymethyl-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidin-2--
ylamino]-phenyl}-piperazin-1-yl)-ethanone
##STR00317##
[0317] By repeating the procedures described in example 102, using
-Chloro-6-diethoxymethyl-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine
as a starting material, the desired product is obtained.
[0318] MS (ESI)m/z 509 (M+H).sup.+
Example 135
2-[4-(4-Acetyl-piperazin-1-yl)-phenylamino]-7-(1-ethyl-propyl)-7H-pyrrolo[-
2,3-d]pyrimidine-6-carbaldehyde
##STR00318##
[0320] To a solution
1-(4-{4-[6-Diethoxymethyl-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidin-2-
-ylamino]-phenyl}-piperazin-1-yl)-ethanone (178 mg, 0.35 mmol) in
1,4 dioxane (2.8 mL) is added 0.8 mL of concentrated HCl at ambient
temperature. The reaction mixture is stirred at ambient temperature
for 30 mins. The mixture is neutralized with 1 N NaOH aqueous
solution and saturated NaHCO.sub.3 aqueous solution, extracted with
EtOAc. The organic layer is washed with brine, dried
Na.sub.2SO.sub.4, and concentrated under reduced pressure to give
160 mg of the title compound as a yellow solid.
[0321] MS (ESI)m/z 435 (M+H).sup.+.
Example 136
##STR00319##
[0323] A mixture of
2-[4-(4-Acetyl-piperazin-1-yl)-phenylamino]-7-(1-ethyl-propyl)-7H-pyrrolo-
[2,3-d]pyrimidine-6-carbaldehyde (25 mg, 0.057 mmol), methoxyl
amine hydrochloride (20 mg, 0.22 mmol) and 6N HCl (0.03 mL) in EtOH
(1 mL) is stirred at ambient temperature for 6 h. The mixture is
quenched with saturated NaHCO.sub.3 aqueous solution, extracted
with CH.sub.2Cl.sub.2. The organic layer is washed with brine,
dried over Na.sub.2SO.sub.4, and concentrated under reduced
pressure to give the crude product. The crude product is purified
by prep-HPLC to give 12 mg of the title compound as a bright yellow
solid.
[0324] MS (ESI)m/z 464 (M+H).sup.+.
Example 137
7-(1-Ethyl-propyl)-2-[3-fluoro-4-(4-methyl-piperazin-1-yl)-phenylamino]-5,-
7-dihydro-pyrrolo[2,3-d]pyrimidin-6-one
##STR00320##
[0326] To a solution of 5-bromo-2,4-dichloropyrimidine (4.56 g, 20
mmol) in ethanol (9 mL) is added 1-ethylpropylamine (2.6 mL, 22
mmol) and N,N-diisopropylethylamine (7 mL, 40 mmol) at ambient
temperature. The reaction mixture is stirred at ambient temperature
for 16 h and concentrated in vacuo. The residue is purified by
flash chromatography (SiO.sub.2, EtOAc/hexane 3:97 to 30:70) to
give (5-bromo-2-chloro-pyrimidin-4-yl)-(1-ethyl-propyl)-amine.
LCMS: 280 (M+H).sup.+
[0327] .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.1 (s, 1H), 5.24
(d, 1H), 4.1 (m, 1H), 1.58 (m, 4H), 0.93 (t, 6H).
[0328] To a solution of tributyl-((Z)-2-ethoxy-vinyl)-stannane
(4.25 g, 8.8 mmol) in CH.sub.3CN (10 mL) is added
(5-bromo-2-chloro-pyrimidin-4-yl)-(1-ethyl-propyl)-amine (2.25 g, 8
mmol), Et.sub.4NCl (1.33 g, 8 mmol) and Pd(PPh.sub.3).sub.2Cl.sub.2
(280 mg, 0.4 mmol) at ambient temperature. The reaction mixture is
purged with N.sub.2, sealed in a microwave reactor and heated at
100.degree. C. for 20 min. After cooling to room temperature, the
mixture is concentrated in vacuo and the residue is purified by
flash chromatography (SiO.sub.2, EtOAc/hexane 5:95 to 40:60) to
give
[2-Chloro-5-((Z)-2-ethoxy-vinyl)-pyrimidin-4-yl]-(1-ethyl-propyl)-amine.
[0329] LCMS: 270 (M+H).sup.+
[0330] .sup.1H NMR (CDCl.sub.3, 400 MHz). .delta. 8.02 (s, 1H),
6.26 (d, 1H), 5.46 (d, 1H), 4.91 (d, 1H), 4.16 (m, 1H), 3.99 (q,
2H), 1.60-1.69 (m, 2H), 1.43-1.52 (m, 2H), 1.32 (t, 3H), 0.92 (t,
6H)
[0331] To a solution of
[2-chloro-5-((Z)-2-ethoxy-vinyl)-pyrimidin-4-yl]-(1-ethylpropyl)-amine
(1.1 g, 4.07 mmol) in EtOH (8 mL) is added concentrated HCl (0.1
mL) at ambient temperature. The reaction mixture is sealed in a
microwave reactor and heated at 100.degree. C. for 10 min. After
cooling to room temperature, the mixture is concentrated in vacuo
to provide 2-chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine.
The crude product is used as it is. The crude product can be
purified by flash chromatography (SiO.sub.2, EtOAc/Hexane 1:5).
[0332] LCMS: 224 (M+H).sup.+
[0333] .sup.1H NMR (CDCl.sub.3, 400 MHz). .delta. 8.87 (s, 1H),
7.30 (d, 1H), 6.69 (d, 1H), 4.69 (m, 1H), 1.77-1.99 (m, 4H), 0.77
(t, 6H)
[0334] To a mixture of
2-chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine (crude,
4.07 mmol) in t-BuOH (7 mL) was added 2 mL of H.sub.2O at ambient
temperature, then NBS (2.28 g, 12.8 mmol) was added to the orange
color solution. The mixture is stirred at 30.degree. C. for 2.5 h,
then is concentrated and taken up in ethyl acetate, washed with
NaHCO.sub.3 aqueous solution, and brine. The organic portion is
dried with Na.sub.2SO.sub.4, filtered and concentrated to provide
5,5-dibromo-2-chloro-7-(1-ethyl-propyl)-5,7-dihydro-pyrrolo[2,3-d]pyrimid-
in-6-one. The crude product is used as it is.
[0335] LCMS: 398 (M+H).sup.+.
[0336] To a solution of
5,5-dibromo-2-chloro-7-(1-ethyl-propyl)-5,7-dihydro-pyrrolo[2,3-d]pyrimid-
in-6-one (crude, .about.5.3 mmol) in acetic acid (6 mL) and THF (4
mL) is added Zn dust (1.37 g, 21 mmol) at 0.degree. C. The mixture
is stirred at 0.degree. C. for 2 min then warmed up to room
temperature, stirred for 30 min. The mixture is filtered through a
pad of Celite, rinsed with ethyl acetate. The filtrate is
concentrated in vacuo and the residue is purified by flash
chromatography (SiO.sub.2, EtOAc/hexane 5:95 to 40:60) to give
2-chloro-7-(1-ethyl-propyl)-5,7-dihydro-pyrrolo[2,3-d]pyrimidin-6-
-one.
[0337] LCMS: 240 (M+H).sup.+
[0338] .sup.1H NMR (CDCl.sub.3, 400 MHz). .delta. 8.17 (s, 1H),
4.20 (m, 1H), 3.58 (s, 2H), 2.10 (m, 2H), 1.84 (m, 2H), 0.84 (t,
6H).
[0339] To a mixture of
2-chloro-7-(1-ethyl-propyl)-5,7-dihydro-pyrrolo[2,3-d]pyrimidin-6-one
(18 mg, 0.075 mmol) and TsOH (1.12 ml, 0.2 M in 1,4 dioxane) is
added 3-fluoro-4-(4-methylpiperazine)aniline (23.5 mg, 0.1125
mmol), and DMF (0.25 mL) at ambient temperature. The reaction
mixture is sealed in a microwave reactor and heated at 140.degree.
C. for 30 min. The mixture is diluted with EtOAc, washed with
NaHCO.sub.3 aqueous solution and brine. The organics is dried over
Na.sub.2SO.sub.4, filtered and concentrated. The crude product is
purified by preparative HPLC to give 27 mg of
7-(1-ethyl-propyl)-2-[3-fluoro-4-(4-methyl-piperazin-1-yl)-phenylamino]-5-
,7-dihydro-pyrrolo[2,3-d]pyrimidin-6-one as a brown solid.
[0340] LCMS: 413 (M+H).sup.+.
[0341] .sup.1H NMR (DMSO, 400 MHz). .delta. 9.48 (s, 1H), 8.09 (s,
1H), 7.73 (d, 1H), 7.36 (d, 1H), 6.96 (t, 1H), 4.08 (m, 1H), 3.60
(s, 2H), 3.32 (m, 4H), 2.96 (m, 4H), 2.25 (s, 3H), 2.11 (m, 2H),
1.78 (m, 2H), 0.79 (t, 6H).
Examples 138-199
[0342] By repeating the procedures described in example 137, using
appropriate starting materials, the following compounds are
obtained.
TABLE-US-00014 MS found Example Structure (M + 1) 138 ##STR00321##
395 139 ##STR00322## 382 140 ##STR00323## 380 141 ##STR00324## 423
142 ##STR00325## 410 143 ##STR00326## 354 144 ##STR00327## 353 145
##STR00328## 423 146 ##STR00329## 440 147 ##STR00330## 453 148
##STR00331## 441 149 ##STR00332## 365 150 ##STR00333## 393 151
##STR00334## 428 152 ##STR00335## 410 153 ##STR00336## 352 154
##STR00337## 380 155 ##STR00338## 407 156 ##STR00339## 435 157
##STR00340## 393 158 ##STR00341## 380 159 ##STR00342## 396 160
##STR00343## 423 161 ##STR00344## 447 162 ##STR00345## 435 163
##STR00346## 466 164 ##STR00347## 364 165 ##STR00348## 378 166
##STR00349## 500 167 ##STR00350## 478 168 ##STR00351## 382 169
##STR00352## 417 170 ##STR00353## 396 171 ##STR00354## 337 172
##STR00355## 410 173 ##STR00356## 459 174 ##STR00357## 410 175
##STR00358## 459 176 ##STR00359## 426 177 ##STR00360## 417 178
##STR00361## 455 179 ##STR00362## 407 180 ##STR00363## 439 181
##STR00364## 412 182 ##STR00365## 407 183 ##STR00366## 458 184
##STR00367## 409 185 ##STR00368## 438 186 ##STR00369## 417 187
##STR00370## 382 188 ##STR00371## 408 189 ##STR00372## 368 190
##STR00373## 396 191 ##STR00374## 354 192 ##STR00375## 368 193
##STR00376## 436 194 ##STR00377## 451 195 ##STR00378## 466 196
##STR00379## 423 197 ##STR00380## 396 198 ##STR00381## 424 199
##STR00382## 422
Example 200
2-[4-(4-Acetyl-piperazin-1-yl)-phenylamino]-7-(1-ethyl-propyl)-5,5-dimethy-
l-5,7-dihydro-pyrrolo[2,3-d]pyrimidin-6-one
##STR00383##
[0344] To a solution of
2-chloro-7-(1-ethyl-propyl)-5,7-dihydro-pyrrolo[2,3-d]pyrimidin-6-one
(40 mg, 0.17 mmol) in THF (1.5 mL) is added NaH (60% dispersion in
mineral oil, 20 mg, 0.42 mmol) at 0.degree. C. The reaction mixture
is stirred for 30 min and then cooled to 0.degree. C. After the
addition of iodomethane (0.023 mL, 0.37 mmol) at 0.degree. C., the
mixture is stirred for 3 hr. The reaction mixture is quenched with
aqueous ammonium chloride solution and extracted with ethyl
acetate. The organics is washed with aqueous sodium carbonate
solution and brine, dried over anhydrous sodium sulfate, evaporated
in vacuo. The residue is purified by flash chromatography
(SiO.sub.2, EtOAc/Hexane 1:10) to give 20 mg of
2-chloro-7-(1-ethyl-propyl)-5,5-dimethyl-5,7-dihydro-pyrrolo[2,3-d]pyrimi-
din-6-one.
[0345] .sup.1H NMR (CDCl.sub.3, 400 MHz). .delta. 8.11 (s, 1H),
4.18 (m, 1H), 2.14 (m, 2H), 1.80 (m, 2H), 1.42 (s, 6H), 0.82 (t,
6H).
[0346] To a solution of
2-chloro-7-(1-ethyl-propyl)-5,5-dimethyl-5,7-dihydro-pyrrolo[2,3-d]pyrimi-
din-6-one (20 mg, 0.075 mmol) in 1,4-dioxane (1 mL) and DMF (0.2
mL) are added 1-[4-(4-amino-phenyl)-piperazin-1-yl]ethanone (24.5
mg, 0.11 mmol) and p-toluenesulfonic acid (17 mg, 0.089 mmol). The
reaction mixture is sealed in a microwave reactor and heated at
140.degree. C. for 30 min. The mixture is diluted with EtOAc and
washed with 1N NaOH solution. The organics is dried over
Na.sub.2SO.sub.4, filtered, and concentrated. The residue is
purified by prep-HPLC to give 30 mg of
2-[4-(4-acetyl-piperazin-1-yl)-phenylamino]-7-(1-ethyl-propyl)-5,5-dimeth-
yl-5,7-dihydro-pyrrolo[2,3-d]pyrimidin-6-one as a pale yellow
solid.
[0347] LCMS: 451 (M+H).sup.+
[0348] .sup.1H NMR (CDCl.sub.3, 400 MHz). .delta. 7.95 (s, 1H),
7.49 (d, 2H), 6.93 (d, 2H), 6.91 (br s, 1H), 4.15 (m, 1H), 3.78 (t,
2H), 3.63 (t, 2H), 3.13 (m, 4H), 2.19 (m, 2H), 2.14 (s, 3H), 1.77
(m, 2H), 1.38 (s, 6H), 0.83 (t, 3H).
Example 201
1-(1-{4-[7-(1-Ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino]-phenyl}--
piperidin-4-yl)-ethanone
##STR00384##
[0350] To a mixture of
1-[4-(4-amino-phenyl)-piperazin-1-yl]ethanone (70.5 mg, 0.32 mmol)
and sodium tert-butoxide (38.4 mg, 0.4 mmol) in 1,4-dioxane (0.3
mL) are added a solution of
2-chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine (60 mg,
0.26 mmol) in 1,4-dioxane (0.6 mL), Pd.sub.2(dba).sub.3 (12.2 mg,
0.013 mmol) and BINAP (16.6 mg, 0.026 mmol). The mixture is
degassed, and heated at 100.degree. C. for 3 h. The mixture is
cooled to room temperature, diluted with EtOAc, and filtered
through a pad of Celite. The filtrate is concentrated under reduced
pressure. The crude product is purified by preparative HPLC to give
84.9 mg of
1-(1-{4-[7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino]-phenyl}-
-piperidin-4-yl)-ethanone as a pale white solid.
[0351] LCMS: 407.3 (M+H).sup.+
[0352] .sup.1H NMR (CDCl.sub.3, 400 MHz). .delta. 8.59 (s, 1H),
7.66 (d, 2H), 7.25 (br s, 1H), 6.97 (d, 2H), 6.96 (d, 1H), 6.44 (d,
1H), 4.50 (m, 1H), 3.81 (t, 2H), 3.65 (t, 2H), 3.14 (m, 4H), 2.17
(s, 3H), 1.90 (m, 4H), 0.82 (t, 6H).
Example 202
[7-(1-Ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-(4-piperazin-1-yl-phe-
nyl)-amine
##STR00385##
[0354] To a mixture of 4-(4-amino-phenyl)-piperazine-1-carboxylic
acid tert-butyl ester (133 mg, 0.48 mmol) and sodium tert-butoxide
(57.6 mg, 0.6 mmol) in 1,4-dioxane (0.5 mL) is added a solution of
2-chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine (90 mg, 0.4
mmol) in 1,4-dioxane (1.0 mL), Pd.sub.2(dba).sub.3 (18.3 mg, 0.02
mmol) and BINAP (25 mg, 0.04 mmol). The mixture is degassed, and
heated at 100.degree. C. for 3 h. The mixture is cooled to room
temperature, diluted with EtOAc, and filtered through celite. The
filtrate is concentrated under reduced pressure. The residue is
purified by flash chromatography (SiO.sub.2, EtOAc:Hexane=1:1) to
give 167 mg of
4-{4-[7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino]-phenyl}-pi-
perazine-1-carboxylic acid tert-butyl ester as a pale yellow
solid.
[0355] LCMS: 465.5 (M+H).sup.+
[0356] To a solution of
4-{4-[7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino]-phenyl}-pi-
perazine-1-carboxylic acid tert-butyl ester (167 mg, 0.36 mmol) in
dichloromethane (3 mL) is added trifluoroacetic acid (1 mL). The
reaction mixture is stirred for 1 h and concentrated in vacuo. The
residue is diluted with dichloromethane, washed with NaHCO.sub.3
solution, dried over Na.sub.2SO.sub.4, and concentrated in vacuo.
Purification by preparative HPLC afforded 130 mg of
[7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-(4-piperazin-1-yl-ph-
enyl)-amine as a yellowish solid.
[0357] LCMS: 365.2 (M+H).sup.+
Examples 203-262
[0358] By repeating the procedures described in example 201 and
202, using appropriate starting materials, the following compounds
are obtained.
TABLE-US-00015 MS found Example Structure (M + 1) 203 ##STR00386##
455.2 204 ##STR00387## 394 205 ##STR00388## 395 206 ##STR00389##
408 207 ##STR00390## 380 208 ##STR00391## 397 209 ##STR00392## 417
210 ##STR00393## 273 211 ##STR00394## 364 212 ##STR00395## 394 213
##STR00396## 391 214 ##STR00397## 406 215 ##STR00398## 431 216
##STR00399## 406 217 ##STR00400## 392 218 ##STR00401## 245 219
##STR00402## 437.4 220 ##STR00403## 423.4 221 ##STR00404## 395.3
222 ##STR00405## 393.2 223 ##STR00406## 465.3 224 ##STR00407##
465.4 225 ##STR00408## 433.3 226 ##STR00409## 433.2 227
##STR00410## 422.4 228 ##STR00411## 421.4 229 ##STR00412## 422.4
230 ##STR00413## 351.2 231 ##STR00414## 423.2 232 ##STR00415##
391.2 233 ##STR00416## 380.3 234 ##STR00417## 380.3 235
##STR00418## 379.2 236 ##STR00419## 433.3 237 ##STR00420## 421.3
238 ##STR00421## 419.4 239 ##STR00422## 377.4 240 ##STR00423##
391.3 241 ##STR00424## 433.4 242 ##STR00425## 391.3 243
##STR00426## 392.3 244 ##STR00427## 393.3 245 ##STR00428## 429.2
246 ##STR00429## 377.2 247 ##STR00430## 379.3 248 ##STR00431##
407.3 249 ##STR00432## 420.5 250 ##STR00433## 393.2 251
##STR00434## 364 252 ##STR00435## 406 253 ##STR00436## 377 254
##STR00437## 405 255 ##STR00438## 378 256 ##STR00439## 363.23 257
##STR00440## 319.16 258 ##STR00441## 346.17 259 ##STR00442## 294.17
260 ##STR00443## 203.17 261 ##STR00444## 350.23 262 ##STR00445##
322.20
Example 263
1-(4-{4-[7-(1-Ethyl-propyl)-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino]-
-phenyl}-piperazin-1-yl)-ethanone
##STR00446##
[0359] To a solution of 3-pentanone (1 g, 11.6 mmol) in anhydrous
1,2-dichloroethane (45 mL) is added allylamine (0.872 mL, 11.6
mmol) followed by NaBH(OAc).sub.3 (3.44 g, 16.3 mmol) at ambient
temperature. The reaction mixture is stirred at room temperature
overnight. The reaction mixture is quenched with 1N NaOH and
extracted with dichloromethane. The extract is dried over
Na.sub.2SO.sub.4 and concentrated in vacuo to give 1.27 g of
allyl-(1-ethyl-propyl)-amine. The crude product is used as it
is.
[0360] To a solution of allyl-(1-ethyl-propyl)-amine (1.27 g, 10
mmol) is added anhydrous isopropanol (50 mL) and
5-bromo-2,4-dichloropyrimidine (3.0 g, 5 mmol) and
diisopropylethylamine (2.61 mL, 15 mmol) at ambient temperature.
The reaction mixture is stirred overnight and concentrated in
vacuo. The residue is purified with column chromatography
(SiO.sub.2, EtOAC/hexane 1:5) to give 2.76 g of
allyl-(5-bromo-2-chloro-pyrimidin-4-yl)-(1-ethyl-propyl)-amine.
[0361] LCMS: 320.0 (M+H).sup.+
[0362] To a solution of
allyl-(5-bromo-2-chloro-pyrimidin-4-yl)-(1-ethyl-propyl)-amine
(2.76 g, 8.7 mmol) in anhydrous DMF (15 mL) is added Pd(OAc).sub.2
(156 mg, 0.69 mmol) and PPh.sub.3 (182 mg, 0.69 mmol) and
triethylamine (2.4 mL, 17.3 mmol). The reaction mixture is stirred
at 100.degree. C. overnight. The reaction mixture is diluted with
EtOAc, washed with water, dried over Na.sub.2SO.sub.4, and
concentrated in vacuo. Purification with column chromatography
(SiO.sub.2, EtOAC/hexane 1:2) gives 0.95 g of
2-chloro-7-(1-ethyl-propyl)-5-methyl-7H-pyrrolo[2,3-d]pyrimidine.
[0363] LCMS: 238.2 (M+H).sup.+
[0364] By repeating the procedures described in example 65, using
2-chloro-7-(1-ethyl-propyl)-5-methyl-7H-pyrrolo[2,3-d]pyrimidine as
a starting material,
1-(4-{-4-[7-(1-ethyl-propyl)-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamin-
o]-phenyl}-piperazin-1-yl)-ethanone is obtained.
[0365] LCMS: 421.2 (M+H).sup.+
Examples 264-319
[0366] By repeating the procedures described in example 115, using
appropriate starting materials, the following compounds are
obtained.
TABLE-US-00016 MS found Example Structure (M + 1) 264 ##STR00447##
393.3 265 ##STR00448## 419.26 266 ##STR00449## 377.24 267
##STR00450## 377.24 268 ##STR00451## 391.26 269 ##STR00452## 405.28
270 ##STR00453## 363.23 271 ##STR00454## 350.21 272 ##STR00455##
392.26 273 ##STR00456## 385 274 ##STR00457## 433.28 275
##STR00458## 391.22 276 ##STR00459## 349.21 277 ##STR00460## 293.2
278 ##STR00461## 376.6 279 ##STR00462## 349.2 280 ##STR00463##
347.20 281 ##STR00464## 333.2 282 ##STR00465## 333.2 283
##STR00466## 378.24 284 ##STR00467## 377.24 285 ##STR00468## 395
286 ##STR00469## 455.17 287 ##STR00470## 456.1 288 ##STR00471##
372.07 289 ##STR00472## 441.21 290 ##STR00473## 405.24 291
##STR00474## 406.26 292 ##STR00475## 406.22 293 ##STR00476## 323.2
294 ##STR00477## 323.2 295 ##STR00478## 338.1 296 ##STR00479##
295.2 297 ##STR00480## 308.2 298 ##STR00481## 294.2 299
##STR00482## 327.1 300 ##STR00483## 294.2 301 ##STR00484## 318.2
302 ##STR00485## 379.22 303 ##STR00486## 344.2 304 ##STR00487##
295.2 305 ##STR00488## 378.2 306 ##STR00489## 315.2 307
##STR00490## 288.28 308 ##STR00491## 286.31 309 ##STR00492## 308.03
310 ##STR00493## 364.2 311 ##STR00494## 336.2 312 ##STR00495##
359.2 313 ##STR00496## 392.25 314 ##STR00497## 391.26 315
##STR00498## 408.24 316 ##STR00499## 407.26 317 ##STR00500## 408.25
318 ##STR00501## 409.23 319 ##STR00502## 406.3
Example 320
(5-Methyl-7-pyridin-2-yl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-(5-piperazin-1-y-
l-pyridin-2-yl)-amine
##STR00503##
[0368] A solution of 5-bromo-2,4-dichloropyrimidine (5.0 g, 22
mmol), allylamine (1.98 mL, 26.4 mmol), and diisopropylethylamine
(5.6 mL, 33.0 mmol) are stirred in ethanol (100 mL) at 50.degree.
C. overnight. The solvent is removed in vacuo and the residue is
partitioned between ethyl acetate and saturated aqueous ammonium
chloride solution. The organic layer is washed with brine, dried
over anhydrous sodium sulfate and evaporated to provide
Allyl-(5-bromo-2-chloro-pyrimidin-4-yl)-amine as a white
crystalline solid (89%), which is used without further
purification.
[0369] A mixture of Allyl-(5-bromo-2-chloro-pyrimidin-4-yl)-amine
(1 g, 4 mmol), palladium(H) acetate, (90 mg, 0.40 mmol),
triphenylphosphine (211 mg, 0.80 mmol) and triethylamine (1.1 mL,
8.0 mmol) in DMF (10 mL) is heated at 100.degree. C. overnight.
After cooling to room temperature the reaction mixture is diluted
with ethyl acetate and washed with brine. The organic phase is
dried (anhydrous Na.sub.2SO.sub.4) and the solvent is evaporated.
The crude product is purified by flash chromatography (gradient
elution EtOAC:Heptanes 0:0 to 1:1) on silicagel to afford
2-Chloro-5-methyl-7H-pyrrolo[2,3-d]pyrimidine as a white solid
(40%).
[0370] A mixture of 2-Chloro-5-methyl-7H-pyrrolo[2,3-d]pyrimidine
(80 mg, 0.48 mmol), 2-bromopyridine (113 mg, 0.72 mmol), copper(I)
iodide (9.1 mg, 0.48 mmol), K.sub.3PO.sub.4(2.02 g, 23.84 mmol),
and trans-1,2-diaminocyclohexane (5.44 mg, 0.48 mmol) in
1,4-dioxane (7 mL) is stirred at 90.degree. C. for 1.5 hrs. After
cooling to room temperature the reaction mixture is diluted with
ethyl acetate and washed with brine. The organic phase is dried
(anhydrous Na.sub.2SO.sub.4) and the solvent is evaporated. The
crude product is purified by flash chromatography (gradient elution
ethyl acetate:hetanes 0:1 to 1:4) on silicagel to afford
2-Chloro-5-methyl-7-pyridin-2-yl-7H-pyrrolo[2,3-d]pyrimidine as a
white solid (55%).
[0371] A mixture of
2-Chloro-5-methyl-7-pyridin-2-yl-7H-pyrrolo[2,3-d]pyrimidine (15
mg, 0.06 mmol), 4-(6-Amino-pyridin-3-yl)-piperazine-1-carboxylic
acid tert-butyl ester (20.5 mg, 0.075 mmol), Pd.sub.2(dba).sub.3
(2.8 mg, 0.0031 mmol), BINAP (3.82 mg, 0.0061 mmol), sodium
tert-butoxide (8.84 mg, 0.092 mmol), and 1,4-dioxane (4 mL) under
nitrogen is heated in a sealed tube apparatus at 100.degree. C. for
2.5 h. After cooling to room temperature the reaction mixture is
diluted with ethyl acetate and washed with brine. The organic phase
is dried (anhydrous Na.sub.2SO.sub.4) and the solvent is
evaporated. The crude product is dissolved in DCM (2 mL) and TFA
(0.5 ml was added). The solution is stirred at room temperature.
The reaction mixture is diluted with ethyl acetate and washed with
brine. The organic phase is dried (anhydrous Na.sub.2SO.sub.4) and
the solvent is evaporated. The crude product is purified by hplc to
afford
(5-Methyl-7-pyridin-2-yl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-(5-piperazin-1--
yl-pyridin-2-yl)-amine as a pale yellow solid (27%, two steps).
.sup.1HNMR (400 MHz, DMSO-d6) 8.80-8.81 (m, 2H), 8.51 (s, 1H), 8.12
(d, J=5.0 Hz, 1H), 8.0-8.11 (m, 1H), 8.0 (d, J=2.9 Hz, 1H), 7.90
(s, 1H), 7.49 (d, J=2.4 Hz, 1H), 7.32 (d, J=2.4 Hz, 1H), 3.04-3.06
(m, 4H), (2.86-2.89 (m, 4H), 2.32 (s, 3H). MS (ESI) m/z 387.09
[M+H].sup.+.
Examples 321-325
[0372] By repeating the procedure described in example 320, using
2-Chloro-5-methyl-7H-pyrrolo[2,3-d]pyrimidine and appropriate
starting materials, the following compounds are prepared.
TABLE-US-00017 MS found Example Structure (M + 1) 321 ##STR00504##
393.16 322 ##STR00505## 386.11 323 ##STR00506## 393.03 324
##STR00507## 376.04 325 ##STR00508## 366
Example 326
1-[7-cyclopentyl-2-(4-piperazin-1-yl-phenylamino)-7H-pyrrolo[2,3-d]pyrimid-
in-5-yl]-ethanone
##STR00509##
[0373] 2-chloro-7-(cyclopentyl)-7H-pyrrolo[2,3-d]pyrimidine is
prepared from cyclopentyl amine and 5-bromo-2,4-dichloropyrimidine
using a method similar to that for the preparation of
2-chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine given in
example 1.
[0374] To a solution of aluminium chloride (400 mg, 2.99 mmol) and
acetyl chloride (711 uL, 10 mmol) in dichloromethane (2 mL) is
added 2-chloro-7-(cyclopentyl)-7H-pyrrolo[2,3-d]pyrimidine (221 mg,
1.0 mmol) in dichloromethane (5 mL), dropwise. After 20 minutes
saturated aqueous sodium bicarbonate is added to pH9-10 and the
solution is extracted with dichloromethane. The organic phase is
dried, anhydrous Na.sub.2SO.sub.4 and concentrated to obtain
1-(2-Chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-ethanone
(255 mg, 97%) as an off-white amorphous solid (97%). .sup.1H-NMR
and LCMS.
[0375]
1-[7-cyclopentyl-2-(4-piperazin-1-yl-phenylamino)-7H-pyrrolo[2,3-d]-
pyrimidin-5-yl]-ethanone is prepared from
1-(2-Chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-ethanone
and 4-(4-Aminophenyl)-piperazine-1-carboxylic acid tert-butyl ester
using a method similar to that for the preparation of Example 202.
[M+H] 405.2.
Example 327
(7-Cyclopentyl-5-isopropyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-(5-piperazin-1-
-yl-pyridin-2-yl)-amine
##STR00510##
[0377]
2-Chloro-7-cyclopentyl-5-isopropyl-7H-pyrrolo[2,3-d]pyrimidine is
prepared from 2-chloro-7-(cyclopentyl)-7H-pyrrolo[2,3-d]pyrimidine
and 2-chloropropane using a method similar to that for the
preparation of
1-(2-Chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-ethanone
given in Example 325.
[0378]
(7-Cyclopentyl-5-isopropyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-(5-pipe-
razin-1-yl-pyridin-2-yl)-amine is prepared from
2-Chloro-7-cyclopentyl-5-isopropyl-7H-pyrrolo[2,3-d]pyrimidine and
4-(6-Amino-pyridin-3-yl)-piperazine-1-carboxylic acid tert-butyl
ester using a method similar to that for the preparation of Example
202. [MH+] 406.21
TABLE-US-00018 MS found Example Structure (M + 1) 328 ##STR00511##
391.26
Example 329
[7-(1-Ethyl-propyl)-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-(4-piperazin-
-1-yl-phenyl)-amine
##STR00512##
[0380] To a microwave vial is added a solution of
(5-bromo-2-chloro-pyrimidin-4-yl)-(1-ethyl-propyl)-amine (0.5 g,
1.80 mmol) in anhydrous toluene (10 mL), tributyl(1-propynyl)-tin
(1.1 mL, 3.6 mmol) and Pd(PPh.sub.3).sub.4 (41.5 mg, 0.036 mmol).
The reaction mixture is heated at 120.degree. C. for 1 h by
employing microwave. The reaction mixture is diluted with EtOAc,
washed with NaHCO.sub.3 aqueous solution and water, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. Purification with
column chromatography (SiO.sub.2, EtOAC/hexane 1:5) gave 0.32 g of
(2-chloro-5-prop-1-ynyl-pyrimidin-4-yl)-(1-ethyl-propyl)-amine
[0381] LCMS: 238.2 (M+H).sup.+
[0382] To a microwave vial are added
(2-chloro-5-prop-1-ynyl-pyrimidin-4-yl)-(1-ethyl-propyl)-amine
(0.22 g, 0.92 mmol), anhydrous DMF (3 mL), and CuI (53 mg, 0.27
mmol). The reaction is heated at 160.degree. C. for 1 h by
employing microwave. The reaction mixture is diluted with EtOAc,
washed with NaHCO.sub.3 aqueous solution and water, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. Purification with
column chromatography (SiO.sub.2, EtOAC/hexane 1:4) gives 43 mg of
2-chloro-7-(1-ethyl-propyl)-6-methyl-7H-pyrrolo[2,3-d]pyrimidine.
[0383] LCMS: 238.2 (M+H).sup.+
[0384] By repeating the procedures described in example 202, using
2-chloro-7-(1-ethyl-propyl)-6-methyl-7H-pyrrolo[2,3-d]pyrimidine as
a starting material,
[7-(1-ethyl-propyl)-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-(4-piperazi-
n-1-yl-phenyl)-amine is obtained.
[0385] LCMS: 379.1 (M+H).sup.+
Examples 330-332
[0386] By repeating the procedures described in example 329, using
appropriate starting materials, the following compounds are
obtained.
TABLE-US-00019 MS found Example Structure (M + 1) 330 ##STR00513##
421.4 331 ##STR00514## 406.3 332 ##STR00515## 377.1
Example 333
1-[7-Cyclopentyl-6-methyl-2-(4-piperazin-1-yl-phenylamino)-7H-pyrrolo[2,3--
d]pyrimidin-5-yl]-ethanone
##STR00516##
[0387] 2-Chloro-7-cyclopentyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidine
is prepared from cyclopentyl amine and
5-bromo-2,4-dichloropyrimidine using a method similar to that for
the preparation of
2-chloro-7-(1-ethyl-propyl)-6-methyl-7H-pyrrolo[2,3-d]pyrimidine
given in example 328.
[0388]
51-(2-Chloro-7-cyclopentyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl-
)-ethanone is prepared from
2-Chloro-7-cyclopentyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidine and
acetyl chloride using a method similar to that for the preparation
of
1-(2-Chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-ethanone
given in Example 325.
[0389]
1-[7-Cyclopentyl-6-methyl-2-(4-piperazin-1-yl-phenylamino)-7H-pyrro-
lo[2,3-d]pyrimidin-5-yl]-ethanone is prepared from
1-(2-Chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-ethanone
and 4-(4-aminophenyl)piperazine-1-carboxylic acid tert-butyl ester
using a method similar to that given for the preparation of Example
202. (MH+) 419.2
Example 334
(5-chloro-7-cyclopentyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-(4-piper-
azin-1-yl-phenyl)-amine
##STR00517##
[0390] To a solution of
2-Chloro-7-cyclopentyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidine (164
mg, 0.70 mmol) in dichloromethane (3 mL) is added
N-chlorosuccinimide (0.4M in DCM, 1.1 eq) over 1 h. The reaction
mixture is allowed to stir for 3 days at room temperature. The
reaction mixture is diluted with dichloromethane and washed with
saturated aqueous sodium bicarbonate followed by brine. The organic
phase is concentrated and the crude product is purified by normal
phase chromatography (SiO.sub.2, EtOAc/heptane) to obtain
2,5-Dichloro-7-cyclopentyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidine
(158 mg, 84%).
[0391]
(5-chloro-7-cyclopentyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-(-
4-piperazin-1-yl-phenyl)-amine is prepared from
2,5-Dichloro-7-cyclopentyl-6-methyl-7H-pyrrolo[2,3-d] and
4-(6-Amino-pyridin-3-yl)-piperazine-1-carboxylic acid tert-butyl
ester using a method similar to that for the preparation of Example
202. (MH+) 412.2
Example 335
7-(1-ethyl-propyl)-2-(4-piperazin-1-yl-phenylamino)-7H-pyrrolo[2,3-d]pyrim-
idine-6-carboxylic acid dimethyl amide
##STR00518##
[0392] To a mixture of
(5-bromo-2-chloro-pyrimidin-4-yl)-(1-ethyl-propyl)-amine (420 mg,
1.5 mmol) and propargylaldehyde diethyl acetal (0.32 mL, 2.25 mmol)
in DMF (6 mL) is added PdCl.sub.2(PPh.sub.3).sub.2 (105 mg, 0.15
mmol) and CuI (28 mg, 0.15 mmol), followed by Et.sub.3N (0.42 mL, 3
mmol). The mixture is degassed, and heated at 55.degree. C. for 16
h. After cooling to room temperature, the reaction mixture is
diluted with EtOAc, washed with water and brine. The organic layer
is dried over Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure. The crude product is purified by column
chromatography (SiO.sub.2, EtOAc/heptane 5:95 to 40:60) to give 182
mg of
[2-chloro-5-(3,3-diethoxy-prop-1-ynyl)-pyrimidin-4-yl]-(1-ethyl-propyl)-a-
mine as a light brown oil.
[0393] LCMS: 326 (M+H).sup.+
[0394] To a solution of
[2-chloro-5-(3,3-diethoxy-prop-1-ynyl)-pyrimidin-4-yl]-(1-ethyl-propyl)-a-
mine (326 mg, 1 mmol) in THF (2 mL) is added 1N TBAF in THF (5 mL,
5 mmol) at ambient temperature. The reaction mixture is heated at
70.degree. C. for 2 h. After cooling down, the mixture is
concentrated in vacuo and purified by BIOTAGE column (EtOAc/heptane
5:5 to 40:60) to give 307 mg of
2-chloro-6-diethoxymethyl-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine
as a light yellow oil.
[0395] LCMS: 326 (M+H).sup.+.
[0396] To a solution of
2-chloro-6-diethoxymethyl-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine
(67 mg, 0.2 mmol) in 1,4-dioxane (0.7 mL) is added conc. HCl (0.2
mL) at ambient temperature. The reaction mixture is stirred for 30
min, then neutralized with 2N NaOH aqueous solution and saturated
NaHCO.sub.3 aqueous solution. The mixture is extracted with EtOAc.
The extracts are washed with brine, dried over Na.sub.2SO.sub.4,
filtered and concentrated in vacuo to give 54 mg of
2-chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine-6-carbaldehyde
as a yellow solid. The crude product is used as it is.
[0397] LCMS: 252 (M+H).sup.+.
[0398] To a mixture of
2-chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine-6-carbaldehyde
(283 mg, 1.11 mmol) in DMF (3 mL) is added oxone (820 mg, 1.33
mmol) at room temperature. The mixture is stirred at room
temperature for 5 h and is quenched with 20% Na.sub.2S.sub.2O.sub.3
aqueous solution. After stirring for 10 min, the reaction mixture
is acidified with 1N HCl aqueous solution (pH=5). The mixture is
extracted with dichloromethane, dried over Na.sub.2SO.sub.4 and
concentrated in vacuo. The solid is filtered, washed with
acetonitrile, and dried under vacuum to give 130 mg of
2-chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic
acid as a pale brown solid.
[0399] LCMS: 268 (M+H).sup.+.
[0400] To a solution of
2-chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic
acid (80 mg, 0.30 mmol), BOP (159 mg, 0.36 mmol) and
N,N-diisopropylethylamine (0.078 mL, 0.45 mmol) in DMF (3 mL) is
added 0.164 mL of 2 N dimethylamine in THF solution at room
temperature. The mixture is stirred at room temperature for 3 h,
quenched with 1N NaOH aqueous solution, and extracted with EtOAc.
The organic extracts are washed with brine, dried over
Na.sub.2SO.sub.4, filtered, and concentrated under reduced
pressure. The crude product is purified by column chromatography
(SiO.sub.2, 5% MeOH in CH.sub.2Cl.sub.2) to give 64 mg of
2-chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic
acid dimethylamide.
[0401] LCMS: 295.1 (M+H).sup.+.
[0402] By repeating the procedures described in example 202, using
2-chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic
acid dimethylamide as a starting material,
7-(1-ethyl-propyl)-2-(4-piperazin-1-yl-phenylamino)-7H-pyrrolo[2,3-d]pyri-
midine-6-carboxylic acid dimethyl amide is obtained.
[0403] LCMS: 436.3 (M+H).sup.+.
Examples 336-359
[0404] By repeating the procedures described in example 335, using
appropriate starting materials, the following compounds are
obtained.
TABLE-US-00020 MS found Example Structure (M + 1) 336 ##STR00519##
464.3 337 ##STR00520## 462.3 338 ##STR00521## 434.27 339
##STR00522## 448.28 340 ##STR00523## 449.26 341 ##STR00524## 433.27
342 ##STR00525## 450.30 343 ##STR00526## 478.3 344 ##STR00527##
476.3 345 ##STR00528## 484.3 346 ##STR00529## 422.2 347
##STR00530## 514.3 348 ##STR00531## 498.3 349 ##STR00532## 540.3
350 ##STR00533## 552.3 351 ##STR00534## 485.3 352 ##STR00535##
485.3 353 ##STR00536## 485.3 354 ##STR00537## 596.4 355
##STR00538## 502.4 356 ##STR00539## 474.2 357 ##STR00540## 502.3
358 ##STR00541## 540.3 359 ##STR00542## 552.3
Example 360
1-(4-{4-[6-Diethoxymethyl-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidin-2--
ylamino]-phenyl}-piperazin-1-yl)-ethanone
##STR00543##
[0405] By repeating the procedures described in example 201, using
2-chloro-6-diethoxymethyl-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine
as a starting material,
1-(4-{4-[6-diethoxymethyl-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidin-2-
-ylamino]-phenyl}-piperazin-1-yl)-ethanone is obtained.
[0406] LCMS: 509 (M+H).sup.+
Example 361
2-[4-(4-Acetyl-piperazin-1-yl)-phenylamino]-7-(1-ethyl-propyl)-7H-pyrrolo[-
2,3-d]pyrimidine-6-carbaldehyde
##STR00544##
[0407] To a solution
1-(4-{4-[6-diethoxymethyl-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidin-2-
-ylamino]-phenyl}-piperazin-1-yl)-ethanone (0.178 g, 0.35 mmol) in
1,4 dioxane (2.8 mL) is added 0.8 mL of concentrated HCl at ambient
temperature. The reaction mixture is stirred at ambient temperature
for 30 min. The mixture is neutralized with 1 N NaOH aqueous
solution and saturated NaHCO.sub.3 aqueous solution, extracted with
CH.sub.2Cl.sub.2. The extracts are washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated under reduced pressure to give
160 mg of
2-[4-(4-acetyl-piperazin-1-yl)-phenylamino]-7-(1-ethyl-propyl)-7H-pyrrolo-
[2,3-d]pyrimidine-6-carbaldehyde as a yellow solid.
[0408] LCMS: 435 (M+H).sup.+.
Example 362
1-(4-{4-[7-(1-Ethyl-propyl)-6-hydroxymethyl-7H-pyrrolo[2,3-d]pyrimidin-2-y-
lamino]-phenyl}-piperazin-1-yl)-ethanone
##STR00545##
[0409] To a solution of
2-[4-(4-acetyl-piperazin-1-yl)-phenylamino]-7-(1-ethyl-propyl)-7H-pyrrolo-
[2,3-d]pyrimidine-6-carbaldehyde (20 mg, 0.046 mmol) in MeOH (1 mL)
is added NaBH.sub.4 (3.5 mg, 0.092 mmol). The reaction mixture is
stirred for 1 h and concentrated in vacuo. The residue is purified
by preparative HPLC to give 15 mg of
1-(4-{4-[7-(1-ethyl-propyl)-6-hydroxymethyl-7H-pyrrolo[2,3-d]pyrimidin-2--
ylamino]-phenyl}-piperazin-1-yl)-ethanone.
[0410] LCMS: 437.3 (M+H).sup.+.
Example 363
1-(4-{4-[7-(1-Ethyl-propyl)-6-oxazol-5-yl-7H-pyrrolo[2,3-d]pyrimidin-2-yla-
mino]-phenyl}-piperazin-1-yl)-ethanone
##STR00546##
[0411] To a solution of
2-[4-(4-acetyl-piperazin-1-yl)-phenylamino]-7-(1-ethyl-propyl)-7H-pyrrolo-
[2,3-d]pyrimidine-6-carbaldehyde (30 mg, 0.07 mmol) in MeOH (1 mL)
are added p-toluenesulfonyl isocyanide (16 mg, 0.08 mmol) and
K.sub.2CO.sub.3 (29 mg, 0.21 mmol). The reaction mixture is heated
at reflux for 1.5 h and concentrated in vacuo. The residue is
purified by preparative HPLC to give 21 mg of
1-(4-{4-[7-(1-ethyl-propyl)-6-oxazol-5-yl-7H-pyrrolo[2,3-d]pyrimidin-2-yl-
amino]-phenyl}-piperazin-1-yl)-ethanone as a pale brown solid.
[0412] LCMS: 474.2 (M+H).sup.+.
Example 364
1-{4-[4-(7-(1-Ethyl-propyl)-6-[1-methoxyimino-ethyl]-7H-pyrrolo[2,3-d]pyri-
midin-2-ylamino)-phenyl]-piperazin-1-yl}-ethanone
##STR00547##
[0413] A mixture of
2-[4-(4-acetyl-piperazin-1-yl)-phenylamino]-7-(1-ethyl-propyl)-7H-pyrrolo-
[2,3-d]pyrimidine-6-carbaldehyde (25 mg, 0.057 mmol), methoxylamine
hydrochloride (20 mg, 0.22 mmol) and 6N HCl (0.03 mL) in EtOH (1
mL) is stirred at ambient temperature for 6 h. The mixture was
quenched with saturated NaHCO.sub.3 aqueous solution, extracted
with CH.sub.2Cl.sub.2. The extracts were washed with brine, dried
over Na.sub.2SO.sub.4, and concentrated under reduced pressure to
give the crude product. The crude product is purified by
preparative HPLC to give 12 mg of
1-{4-[4-(7-(1-ethyl-propyl)-6-{1-methoxyimino-ethyl}-7H-pyrrolo[2,3-d]pyr-
imidin-2-ylamino)-phenyl]-piperazin-1-yl}-ethanone as a bright
yellow solid.
[0414] LCMS: 464 (M+H).sup.+.
Example 365
1-{4-[4-(7-(1-Ethyl-propyl)-6-{[(furan-2-ylmethyl)-amino]-methyl}-7H-pyrro-
lo[2,3-d]pyrimidin-2-ylamino)-phenyl]-piperazin-1-yl}-ethanone
##STR00548##
[0415] To a solution of
2-[4-(4-acetyl-piperazin-1-yl)-phenylamino]-7-(1-ethyl-propyl)-7H-pyrrolo-
[2,3-d]pyrimidine-6-carbaldehyde (30 mg, 0.07 mmol) in THF (1 mL)
is added furfurylamine (0.03 mL, 0.35 mmol), NaBH(OAc).sub.3 (45
mg, 0.21 mmol), and acetic acid (1 mL). The reaction mixture is
stirred for 16 h and concentrated in vacuo. The residue is purified
by preparative HPLC to give 20 mg of
1-{4-[4-(7-(1-ethyl-propyl)-6-{[(furan-2-ylmethyl)-amino]-methyl}-7H-pyrr-
olo[2,3-d]pyrimidin-2-ylamino)-phenyl]-piperazin-1-yl}-ethanone as
a pale yellow solid.
[0416] LCMS: 516.3 (M+H).sup.+.
Examples 366-372
[0417] By repeating the procedures described in example 365, using
appropriate starting materials, the following compounds are
obtained.
TABLE-US-00021 MS found Example Structure (M + 1) 366 ##STR00549##
478.6 367 ##STR00550## 436.4 368 ##STR00551## 579.4 369
##STR00552## 513.3 370 ##STR00553## 504.3 371 ##STR00554## 503.3
372 ##STR00555## 516.6
Example 373
2-[4-(4-Acetyl-piperazin-1-yl)-phenylamino]-7-(1-ethyl-propyl)-7H-pyrrolo[-
2,3-d]pyrimidine-6-carboxylic acid methyl ester
##STR00556##
[0418] To a solution of
2-chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic
acid (13 mg, 0.049 mmol) in MeOH (0.5 mL) is added
(trimethylsilyl)diazomethane (0.07 mL of a 2.0 M in hexanes). The
reaction mixture is stirred for 2 h and concentrated in vacuo to
give 13 mg of
2-chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxyli-
c acid methyl ester. The crude product is used as it is.
[0419] LCMS: 282.2 (M+H).sup.+.
To a solution of 1-[4-(4-amino-phenyl)-piperazin-1-yl]-ethanone
(12.1 mg, 0.055 mmol) in 1,4-dioxane (0.5 mL) is added a solution
of
2-chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic
acid methyl ester (13 mg, 0.046 mmol) in 1,4-dioxane (0.6 mL),
Pd.sub.2(dba).sub.3 (2.2 mg, 0.0023 mmol), Xantphos (2.7 mg, 0.046
mmol) and Cs.sub.2CO.sub.3 (22.5 mg, 0.069 mmol). The mixture is
degassed, and heated at 100.degree. C. for 3 h. The mixture is
cooled to room temperature, diluted with EtOAc, and filtered
through a pad of Celite. The filtrate is concentrated under reduced
pressure. The crude product is purified by preparative HPLC to give
8.6 mg of
2-[4-(4-acetyl-piperazin-1-yl)-phenylamino]-7-(1-ethyl-propyl)-7H-pyrrolo-
[2,3-d]pyrimidine-6-carboxylic acid methyl ester as a pale white
solid.
[0420] LCMS: 465.4 (M+H).sup.+.
Example 374
2-[4-(4-Acetyl-piperazin-1-yl)-phenylamino]-7-(1-ethyl-propyl)-7H-pyrrolo[-
2,3-d]pyrimidine-6-carboxylic acid
##STR00557##
[0421] To a solution of
2-[4-(4-acetyl-piperazin-1-yl)-phenylamino]-7-(1-ethyl-propyl)-7H-pyrrolo-
[2,3-d]pyrimidine-6-carboxylic acid methyl ester (19 mg, 0.041
mmol) in MeOH (1.5 mL) is added 2 N LiOH aqueous solution (0.5 mL).
The reaction mixture is stirred overnight and concentrated in
vacuo. The residue is purified by preparative HPLC to give 13.6 mg
of
2-[4-(4-Acetyl-piperazin-1-yl)-phenylamino]-7-(1-ethyl-propyl)-7H-pyrrolo-
[2,3-d]pyrimidine-6-carboxylic acid.
[0422] LCMS: 451.4 (M+H).sup.+.
Example 375
7-(1-Ethyl-propyl)-2-(4-piperazin-1-yl-phenylamino)-7H-pyrrolo[2,3-d]pyrim-
idine-6-carboxylic acid
##STR00558##
[0423] To a solution of
2-[4-(4-acetyl-piperazin-1-yl)-phenylamino]-7-(1-ethyl-propyl)-7H-pyrrolo-
[2,3-d]pyrimidine-6-carboxylic acid methyl ester (16 mg, 0.034
mmol) in THF (1.5 mL) is added 2 N LiOH aqueous solution (1 mL).
The reaction mixture is stirred for 36 h and concentrated in vacuo.
The residue is purified by preparative HPLC to give 9.4 mg of
741-ethyl-propyl)-2-(4-piperazin-1-yl-phenylamino)-7H-pyrrolo[2,3-d]pyrim-
idine-6-carboxylic acid.
[0424] LCMS: 409.4 (M+H).sup.+.
Example 376
1-(4-{4-[7-(1-Ethyl-propyl)-6-(1-hydroxy-ethyl)-7H-pyrrolo[2,3-d]pyrimidin-
-2-ylamino]-phenyl}-piperazin-1-yl)-ethanone
##STR00559##
[0425] To a mixture of
(5-bromo-2-chloro-pyrimidin-4-yl)-(1-ethyl-propyl)-amine (0.44 g,
1.58 mmol), Pd(Ph.sub.3P).sub.2Cl.sub.2 (0.11 g, 0.16 mmol), and
CuI (0.03 g, 0.16 mmol) in DMF (14 mL) is added 3-butyn-2-ol (0.19
mL, 2.37 mmol) and triethylamine (0.44 mL, 3.16 mmol). The reaction
mixture is stirred at 55.degree. C. for 16 h, diluted with
CH.sub.2Cl.sub.2, filtered through a pad of Celite, and
concentrated in vacuo. Purification by flash chromatography
(SiO.sub.2, EtOAc/Hexane 1:3) gave 0.23 g of
4-[2-chloro-4-(1-ethyl-propylamino)-pyrimidin-5-yl]-but-3-yn-2-ol
as yellowish oil.
[0426] LCMS: 268 (M+H).sup.+.
[0427] To a solution of
4-[2-chloro-4-(1-ethyl-propylamino)-pyrimidin-5-yl]-but-3-yn-2-ol
(0.23 g, 0.85 mmol) in THF (0.5 mL) is added 1M TBAF (4.3 mL). The
reaction mixture is heated at reflux for 16 h, diluted with water,
extracted with EtOAc. The extracts are dried over Na.sub.2SO.sub.4
and concentrated in vacuo. The residue is purified by flash
chromatography (SiO.sub.2, EtOAc/Hexane 1:3) to provide 0.12 g of
1-[2-chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidin-6-yl]ethanol.
[0428] LCMS: 268 (M+H).sup.+.
[0429] By repeating the procedures described in example 65, using
1-[2-chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidin-6-yl]-ethanol
as a starting material,
1-(4-{4-[7-(1-ethyl-propyl)-6-(1-hydroxy-ethyl)-7H-pyrrolo[2,3-d]pyrimidi-
n-2-ylamino]-phenyl}-piperazin-1-yl)-ethanone is obtained.
[0430] LCMS: 451.4 (M+H).sup.+.
Example 377
1-[2-[4-(4-Acetyl-piperazin-1-yl)-phenylamino]-7-(1-ethyl-propyl)-7H-pyrro-
lo[2,3-d]pyrimidin-6-yl]-ethanone
##STR00560##
[0431] To a solution of
1-[2-chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidin-6-yl]ethanol
(61 mg, 0.2 mmol) in CH.sub.2Cl.sub.2 (2 mL) is added Dess-Martin
periodinane (242 mg, 0.5 mmol). The reaction mixture is stirred for
1 h, quenched with 10% NaS.sub.2O.sub.3: saturated NaHCO.sub.3
(1:1) aqueous solution, and extracted with CH.sub.2Cl.sub.2. The
extracts are washed with water and brine, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The residue is
purified by flash chromatography (SiO.sub.2, EtOAc/Hexane 1:3) to
afford 58 mg of
1-[2-chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidin-6-yl]-ethanone.
[0432] LCMS: 266 (M+H).sup.+.
[0433] By repeating the procedures described in example 201, using
1-[2-chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidin-6-yl]-ethanone
as a starting material,
1-[2-[4-(4-acetyl-piperazin-1-yl)-phenylamino]-7-(1-ethyl-propyl)-7H-pyrr-
olo[2,3-d]pyrimidin-6-yl]-ethanone is obtained.
[0434] LCMS: 449.4 (M+H).sup.+.
Example 378
[7-(1-Ethyl-propyl)-6-(1-methoxy-ethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl]-(-
4-piperazin-1-yl-phenyl)-amine
##STR00561##
[0435] To a solution of
1-(4-{4-[7-(1-ethyl-propyl)-6-(1-hydroxy-ethyl)-7H-pyrrolo[2,3-d]pyrimidi-
n-2-ylamino]phenyl}-piperazin-1-yl)-ethanone (19 mg, 0.042 mmol) in
MeOH (1 mL) is added 4N HCl in dioxane (1 mL). The reaction mixture
is stirred at 60.degree. C. for 2 h. The mixture is loaded on Solid
Phase Extraction column (Sorbent: benzensulfonic acid), washed with
MeOH, eluted with EtOAc:MeOH:Et.sub.3N (1:1:0.05), concentrated in
vacuo. The residue is purified by preparative HPLC to give 10 mg of
[7-(1-ethyl-propyl)-6-(1-methoxy-ethyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl]--
(4-piperazin-1-yl-phenyl)-amine.
[0436] LCMS: 423.4 (M+H).sup.+.
Examples 379-382
[0437] By repeating the procedures described in example 378, using
appropriate starting materials, the following compounds are
obtained.
TABLE-US-00022 MS found Example Structure (M + 1) 379 ##STR00562##
451.3 380 ##STR00563## 436.3 381 ##STR00564## 509.4 382
##STR00565## 409.3
Example 383
(7-Cyclopentyl-6-isopropenyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-(4--
piperazin-1-yl-phenyl)-amine
##STR00566##
[0438] (5-Bromo-2-chloro-pyrimidin-4-yl)cyclopentylamine is
prepared from cyclopentyl amine and 5-bromo-2,4-dichloropyrimidine
using a method similar to that for the preparation of
(5-Bromo-2-chloro-pyrimidin-4-yl)-(1-ethylpropyl)amine given in
Example 137.
[0439] To a solution of
(5-Bromo-2-chloro-pyrimidin-4-yl)cyclopentylamine (1 g, 3.616 mmol)
is added lithium chloride (153.7 mg, 3.616 mmol) and potassium
acetate (887.12 mg, 9.03 mmol) in DMF (50 mL). The solution is
degassed and back-filled with N.sub.2. Palladium(II) acetate (40.6
mg, 0.18 mmol) is added and the solution is degassed and
back-filled with nitrogen three times. 3-pentyn-2-ol (1.0 mL, 10.8
mmol) is added and the reaction solution is heated at 120.degree.
C. for 5 hours. LC-MS analysis indicated the absence of starting
material and the formation of a pair of regio-isomeric products.
After cooling to room temperature, the mixture is filtered through
Celite, diluted with water, and extracted with ethyl acetate EtOAc
three times. The organic layers are combined, washed with brine and
dried over anhydrous sodium sulfate. The solvent is evaporated and
the crude material is purified using silica gel chromatography (30%
ethyl acetate/70% hexanes to give
1-(2-Chloro-7-cyclopentyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-6-yl)-ethan-
ol as a pale powder (150 mg, 14.8%). [M+H].sup.+=280.07.
[0440]
1-(2-Chloro-7-cyclopentyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-6-yl)-
-ethanone is prepared by Dess-Martin periodinane oxidation of
1-(2-Chloro-7-cyclopentyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-6-yl)-ethan-
ol using a procedure similar to that described for
1-[2-chloro-7-(1-ethyl-propyl)-7H-pyrrolo[2,3-d]pyrimidin-6-yl]-ethanone
in the synthesis of Example 376. [M+H].sup.+=278.03.
[0441] A solution of
1-(2-Chloro-7-cyclopentyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-6-yl)-ethan-
one (40 mg, 0.144 mmol),
1-[4-(4-Amino-phenyl)-piperazin-1-yl]-ethanone (37.9 mg, 0.172
mmol), Pd.sub.2(dba).sub.3 (6.7 mg, 0.007 mmol), BINAP (9.15 mg,
0.014 mmol) and NaOtBu (20.7 mg, 0.216 mmol) in 1,4-dioxane (4 mL)
is degassed and back-filled with nitrogen three times. The reaction
mixture is heated to 80.degree. C. for 2 hours. After cooling to
room temperature water is added and the reaction mixture is
extracted with ethyl acetate three times. The organic layers are
combined, washed with brine and dried over anhydrous sodium
sulfate. The solvent is evaporated and the crude material is
purified by preparative HPLC to provide
1-{2-[4-(4-Acetyl-piperazin-1-yl)-phenylamino]-7-cyclopentyl-5-methyl-7H--
pyrrolo[2,3-d]pyrimidin-6-yl}-ethanone (16 mg, 24%).
[M+H].sup.+=461.13.
[0442] To a solution of
1-{2-[4-(4-Acetyl-piperazin-1-yl)-phenylamino]-7-cyclopentyl-5-methyl-7H--
pyrrolo[2,3-d]pyrimidin-6-yl}-ethanone (12 mg, 0.026 mmol) in
methanol (3 mL) is added HCl (2 mL, 2M in dioxane), dropwise. The
solution is heated to reflux for 2 h. The solvent is evaporated and
the crude product is purified on HPLC to give a TFA salt of
(7-Cyclopentyl-6-isopropenyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-(4-
-piperazin-1-yl-phenyl)-amine as a yellow solid (7 mg, 42%).
[M+H].sup.+=419.17.
Example 384
7-Cyclopentyl-5-methyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2-
,3-d]pyrimidine-6-carboxylic acid methyl ester
##STR00567##
[0443] A solution of
(5-Bromo-2-chloro-pyrimidin-4-yl)cyclopentylamine (8 g, 28.93
mmol), lithium chloride (1.23 g, 28.9 mmol), potassium carbonate
(10 g, 72 mmol) and palladium acetate (324.68 mg, 1.45 mmol) in DMF
(300 mL) is degassed and back-filled filled with nitrogen three
times. Methyl-2-butynoate (8.5 mL, 87 mmol) is added and the
reaction solution is heated at 120.degree. C. for 5 h. LC-MS
indicated the formation of two regioisomers and no starting
material remaining. After cooling to room temperature the solution
is filtered through Celite, diluted with water and extracted with
ethyl acetate three times. The organic layers are combined, washed
with brine and dried over anhydrous sodium sulfate. The solvent is
evaporated and the crude product is purified using silica gel
chromatography (.about.20% ethyl acetate/80% hexane) to give
2-chloro-7-cyclopentyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic
acid methyl ester as a yellow solid (2.11 g, 25%).
[M+H].sup.+=294.04.
[0444] A mixture of
2-chloro-7-cyclopentyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic
acid methyl ester (110 mg, 0.374 mmol),
4-(6-Amino-pyridin-3-yl)-piperazine-1-carboxylic acid tert-butyl
ester (114.66 mg, 0.412 mmol), Pd.sub.2(dba).sub.3(17.144 mg, 0.02
mmol), Xantphos (21.67 mg, 0.037 mmol) and cesium carbonate (183
mg, 0.562 mmol) in dioxane (5 mL) is degassed and back-filled with
nitrogen three times. The reaction mixture is heated to 100.degree.
C. for 4 h. Water is added and the solution is extracted with ethyl
acetate three times. The organic layers are combined, washed with
brine and dried over anhydrous sodium sulfate. The solvent is
evaporated and the crude product is dissolved in a small amount of
ethyl acetate. White solid precipitates out and filters to give
2-[5-(4-tert-Butoxycarbonyl-piperazin-1-yl)-pyridin-2-ylamino]-7--
cyclopentyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acid
methyl ester as a white solid (35 mg, 17%) which was carried onto
the next step without further purification. [M+H].sup.+=536.35.
[0445] To a solution of
2-[5-(4-tert-Butoxycarbonyl-piperazin-1-yl)-pyridin-2-ylamino]-7-cyclopen-
tyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acid methyl
ester (35 mg, 0.065 mmol) in DCM (8 mL) is added TFA (2 mL)
dropwise. The solution is stirred at room temperature for 2 hours.
Solvent is evaporated and the crude material is purified by
preparative HPLC to give the TFA salt of
7-Cyclopentyl-5-methyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[-
2,3-d]pyrimidine-6-carboxylic acid methyl ester as yellow solid (32
mg, 74%). [M+H].sup.+=436.2458.
Example 385
7-Cyclopentyl-6-methyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2-
,3-d]pyrimidine-5-carboxylic acid methyl ester
##STR00568##
[0446]
2-chloro-7-cyclopentyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidine-5-carbo-
xylic acid methyl ester is prepared using the procedure shown for
the preparation of its regio-isomer
2-chloro-7-cyclopentyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic
acid methyl ester given in the procedure for the synthesis of
Example 384.
7-Cyclopentyl-6-methyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2-
,3-d]pyrimidine-5-carboxylic acid methyl ester is prepared from
2-chloro-7-cyclopentyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic
acid methyl ester using a procedure similar to that given for the
preparation of
7-Cyclopentyl-5-methyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[-
2,3-d]pyrimidine-6-carboxylic acid methyl ester, Example 384.
[0447] [M+H].sup.+=436.25.
Example 386
[7-Cyclopentyl-5-methyl-6-(5-methyl-[1,3,4]oxadiazol-2-yl)-7H-pyrrolo[2,3d-
]pyrimidin-2-yl]-(5-piperazin-1-yl-pyridin-2-yl)-amine
##STR00569##
[0449] To a suspension of
2-[5-(4-tert-Butoxycarbonyl-piperazin-1-yl)-pyridin-2-ylamino]-7-cyclopen-
tyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acid methyl
ester (250 mg, 0.467 mmol) (prepared as described in the procedure
for the synthesis of Example 384), in MeOH/H.sub.2O/DCM (70 mL) is
added a solution of lithium hydroxide (39.2 mg, 0.93 mmol) in water
(15 mL). The reaction mixture is heated to reflux for 4 hours. The
reaction mixture is allowed to cool to room temperature and
concentrated in vacuo. The resulting solution is acidified to
pH.about.3 using saturated aqueous citric acid solution. The
solution is evaporated and the residue is purified by preparative
HPLC to give
2-[5-(4-tert-Butoxycarbonyl-piperazin-1-yl)-pyridin-2-ylamino]-7-cyclopen-
tyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acid as a
yellow solid (105 mg, 53%). [M+H].sup.+=522.3
[0450] To a solution of 12
2-[5-(4-tert-Butoxycarbonyl-piperazin-1-yl)-pyridin-2-ylamino]-7-cyclopen-
tyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acid (120 mg,
0.23 mmol), HBTU (130.9 mg, 0.345 mmol) and HOAt (46.97 mg, 0.345
mmol) in dry DMF (15 mL) is added a solution of acetic hydrazide
(34.09 mg, 0.46 mmol) and diisopropylethylamine (121 ul, 0.693
mmol) in dry DMF (5 mL). The reaction mixture is stirred at room
temperature overnight. The reaction mixture is diluted with water
and extracted with ethyl acetate three times. The organic layers
are combined, washed with brine and dried over anhydrous sodium
sulfate. The solvent is evaporated and the crude product is
purified by preparative HPLC to give
4-{6-[6-(N'-Acetyl-hydrazinocarbonyl)-7-cyclopentyl-5-methyl-7H-pyrrolo[2-
,3-d]pyrimidin-2-ylamino]-pyridin-3-yl}-piperazine-1-carboxylic
acid tert-butyl ester as a yellow solid (120 mg, 75.4%).
[M+H].sup.+=578.32.
[0451] A mixture of
4-{6-[6-(N'-Acetyl-hydrazinocarbonyl)-7-cyclopentyl-5-methyl-7H-pyrrolo[2-
,3-d]pyrimidin-2-ylamino]-pyridin-3-yl}-piperazine-1-carboxylic
acid tert-butyl ester (80 mg, 0.139 mmol) and polyphosphoric acid
(20 mL) os heated to 120.degree. C. for 1 h. The reaction mixture
is diluted with cold water in an ice-water bath and neutralized to
pH.about.8 with 6 N sodium hydroxide solution. The aqueous solution
is extracted with ethyl acetate three times. The organic layers are
combined, washed with brine and dried over anhydrous sodium
sulfate. The solvent is evaporated and the crude product is washed
with MeOH to give
[7-Cyclopentyl-5-methyl-6-(5-methyl-[1,3,4]oxadiazol-2-yl)-7H-pyrrolo[2,3-
d]pyrimidin-2-yl]-(5-piperazin-1-yl-pyridin-2-yl)-amine as a yellow
solid (28 mg). The methanolic solution is purified by preparative
HPLC to give a TFA salt of
[7-Cyclopentyl-5-methyl-6-(5-methyl-[1,3,4]oxadiazol-2-yl)-7H-pyrrolo[2,3-
d]pyrimidin-2-yl]-(5-piperazin-1-yl-pyridin-2-yl)-amine as a yellow
solid (20 mg). [M+H].sup.+=460.2572.
[0452] Examples 387-408 are prepared using methods similar to those
described in the syntheses of Examples 383-386 and standard
synthetic methodology used in the synthesis of azole heterocycles
with appropriate choice of starting materials.
TABLE-US-00023 MS found Example Structure (M + 1) 387 ##STR00570##
420.25 388 ##STR00571## 436 389 ##STR00572## 448.3 390 ##STR00573##
422.23 391 ##STR00574## 421.25 392 ##STR00575## 449.28 393
##STR00576## 403.24 394 ##STR00577## awaited 395 ##STR00578##
Awaited 396 ##STR00579## Awaited 397 ##STR00580## 333.2 398
##STR00581## 444.25 399 ##STR00582## 445.25 400 ##STR00583## 473.3
401 ##STR00584## 459.25 402 ##STR00585## 458.26 403 ##STR00586##
459.26 404 ##STR00587## 458.23 405 ##STR00588## 459.26 406
##STR00589## 459.27 407 ##STR00590## 460.26 408 ##STR00591##
435.25
Example 409
2-[4-(4-Acetyl-piperazin-1-yl)-phenylamino]-9-(1-ethyl-propyl)-7,9-dihydro-
-purin-8-one
##STR00592##
[0453] To a solution of 2,4-dichloro-5-nitro-pyrimidine (2 g, 10.3
mmol) in anhydrous EtOH (20 mL) is added 1-ethylpropylamine (1.3
mL, 11.3 mmol) and N,N-diisopropylethylamine (2.7 mL, 15.5 mmol) at
0.degree. C. The reaction mixture is stirred for 8 h and
concentrated in vacuo. The residue is dissolved with EtOAc, washed
with saturated NaHCO.sub.3 and brine, dried over Na.sub.2SO.sub.4,
and concentrated in vacuo. Purification with column chromatography
(SiO.sub.2, EtOAC/Hexane 1:3) gives 1.5 g of
(2-chloro-5-nitro-pyrimidin-4-yl)-(1-ethyl-propyl)-amine.
[0454] LCMS: 245.1 (M+H).sup.+
[0455] To a solution of
(2-chloro-5-nitro-pyrimidin-4-yl)-(1-ethyl-propyl)-amine (1 g, 4.1
mmol) in anhydrous EtOH (50 mL) is added Tin(II) chloride (2.3 g,
12.3 mmol) and concentrated HCl (1 mL) at ambient temperature. The
reaction is heated to 80.degree. C. for 1 h and quenched with 1N
NaOH at 0.degree. C. The mixture is extracted with EtOAc, washed
with brine, dried over Na.sub.2SO.sub.4, and concentrated in vacuo
to give 0.5 g of
(2-chloro-5-amino-pyrimidin-4-yl)-(1-ethyl-propyl)-amine. The crude
is used as is.
[0456] LCMS: 215.2 (M+H).sup.+
[0457] To a microwave vial is added the crude
(2-chloro-5-amino-pyrimidin-4-yl)-(1-ethyl-propyl)-amine (0.5 g,
2.3 mmol) and anhydrous DMF (15 mL) followed by
1,1'-carbonyldiimidazole (1.1 g, 7.0 mmol). Sealed vial and
microwave are heated at 100.degree. C. for 10 min. The reaction
mixture is diluted with EtOAc, washed with water, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. Purification with
column chromatography (SiO.sub.2, EtOAC/Hexane 1:1) gives 0.3 g of
2-chloro-9-(1-ethyl-propyl)-7,9-dihydro-purin-8-one. LCMS: 241.1
(M+H).sup.+
[0458] To a mixture of
2-chloro-9-(1-ethyl-propyl)-7,9-dihydro-purin-8-one (95 mg, 0.4
mmol) and TsOH (1.6 mL, 0.2 M in 1,4-dioxane) in DMF (0.25 mL) is
added 1-[4-(4-amino-phenyl)-piperazin-1-yl]-ethanone (105 mg, 0.5
mmol). The reaction mixture is sealed in a microwave reactor and
heated at 140.degree. C. for 30 min. The mixture is diluted with
EtOAc and washed with NaHCO.sub.3 aqueous solution and brine. The
organic layer is dried over Na.sub.2SO.sub.4, filtered and
concentrated. The crude product is purified by preparative HPLC to
give 52 mg of
2-[4-(4-acetyl-piperazin-1-yl)-phenylamino]-9-(1-ethyl-propyl)-7,9-dihydr-
o-purin-8-one as a brown solid.
[0459] LCMS: 424.2 (M+H).sup.+
Examples 410-418
[0460] By repeating the procedures described in example x1, using
appropriate starting materials, the following compounds are
obtained.
TABLE-US-00024 MS found Example Structure (M + 1) 410 ##STR00593##
395.5 411 ##STR00594## 382.2 412 ##STR00595## 438.2 413
##STR00596## 383.2 414 ##STR00597## 383.2 415 ##STR00598## 460.2
416 ##STR00599## 411.2 417 ##STR00600## 424.2 418 ##STR00601##
409.2
Example 419
2-[4-(4-Acetyl-piperazin-1-yl)-phenylamino]-9-(1-ethyl-propyl)-7-methyl-7,-
9-dihydro-purin-8-one
##STR00602##
[0461] To a solution of
2-chloro-9-(1-ethyl-propyl)-7,9-dihydro-purin-8-one (100 mg, 0.41
mmol) in anhydrous DMF (2 mL) is added methyl iodide (21 uL, 0.41
mmol) and NaH (50% dispersion in mineral oil, 22 mg, 0.46 mmol).
The reaction is stirred for 1.5 h. The reaction mixture is quenched
with ice water and extracted with EtOAc. The extracts are dried
over Na.sub.2SO.sub.4 and concentrated in vacuo to give 104 mg of
2-chloro-9-(1-ethyl-propyl)-7-methyl-7,9-dihydro-purin-8-one. The
crude product is used as is.
[0462] LCMS: 255.1 (M+H).sup.+
[0463] To a mixture of
2-chloro-9-(1-ethyl-propyl)-7-methyl-7,9-dihydro-purin-8-one (102
mg, 0.4 mmol) and TsOH (1.6 mL, 0.2 M in 1,4-dioxane) in DMF (0.25
mL) is added 1-[4-(4-amino-phenyl)-piperazin-1-yl]-ethanone (105
mg, 0.5 mmol). The reaction mixture is sealed in a microwave
reactor and heated at 140.degree. C. for 30 min. The mixture is
diluted with EtOAc and washed with NaHCO.sub.3 aqueous solution and
brine. The organics are dried over Na.sub.2SO.sub.4, filtered and
concentrated. The crude product is purified by preparative HPLC to
give 50 mg of
2-[4-(4-acetyl-piperazin-1-yl)-phenylamino]-9-(1-ethyl-propyl)-7,9-dihydr-
o-purin-8-one.
[0464] LCMS: 437.6 (M+H).sup.+
Example 420
9-(1-Ethyl-propyl)-7-methyl-2-[4-(4-methyl-piperazin-1-yl)-phenylamino]-7,-
9-dihydro-purin-8-one
##STR00603##
[0465] By repeating the procedures described in example x11, using
appropriate starting materials,
9-(1-ethyl-propyl)-7-methyl-2-[4-(4-methyl-piperazin-1-yl)-phenylamino]-7-
,9-dihydro-purin-8-one is obtained.
[0466] LCMS: 409.5 (M+H).sup.+
Example 421
1-(4-{4-[9-(1-Ethyl-propyl)-9H-purin-2-ylamino]-phenyl}-piperazin-1-yl)-et-
hanone
##STR00604##
[0467] To a solution of
2-chloro-9-(1-ethyl-propyl)-7,9-dihydro-purin-8-one (0.5 g, 2.3
mmol) in DMF (5 mL) is added triethyl orthoformate (3.8 mL, 23
mmol) and TsOH (0.88 g, 2.6 mmol). The reaction is stirred
overnight. The reaction mixture is quenched with ice water and
extracted with EtOAc. The extracts are dried over Na.sub.2SO.sub.4
and concentrated in vacuo. Purification by flash chromatography
(SiO.sub.2, EtOAC/Hexane 1:3) gives 0.39 g of
2-chloro-9-(1-ethyl-propyl)-9H-purine.
[0468] LCMS: 225.1 (M+H).sup.+
[0469] To a mixture of 2-chloro-9-(1-ethyl-propyl)-9H-purine (90
mg, 0.4 mmol) and TsOH (1.6 mL, 0.2 M in 1,4-dioxane) in DMF (0.25
mL) is added 1-[4-(4-amino-phenyl)-piperazin-1-yl]-ethanone (105
mg, 0.5 mmol). The reaction mixture is sealed in a microwave
reactor and heated at 140.degree. C. for 30 min. The mixture is
diluted with EtOAc and washed with NaHCO.sub.3 aqueous solution and
brine. The organics are dried over Na.sub.2SO.sub.4, filtered and
concentrated. The crude product is purified by preparative HPLC to
give 40 mg of
1-(4-{4-[9-(1-ethyl-propyl)-9H-purin-2-ylamino]-phenyl}-piperazin-1-yl)-e-
thanone.
[0470] LCMS: 407.5 (M+H).sup.+
Example 422
[9-(1-Ethyl-propyl)-9H-purin-2-yl]-[4-(4-methyl-piperazin-1-yl)-phenyl]-am-
ine
##STR00605##
[0471] By repeating the procedures described in example x13, using
appropriate starting materials,
[9-(1-ethyl-propyl)-9H-purin-2-yl]-[4-(4-methyl-piperazin-1-yl)-phenyl]-a-
mine is obtained.
[0472] LCMS: 379.5 (M+H).sup.+
[0473] The following examples of compounds were also produced using
the materials and methods as described above.
TABLE-US-00025 Example Structure Number MS ##STR00606## 423 464
##STR00607## 424 364 ##STR00608## 425 492.3075 ##STR00609## 426
392.2561 ##STR00610## 427 463.2818 ##STR00611## 428 ##STR00612##
429 ##STR00613## 430 377.1 ##STR00614## 431 478.2925 ##STR00615##
432 442.1345 ##STR00616## 433 398.1864 ##STR00617## 436 486.22
##STR00618## 437 385.2 ##STR00619## 438 391.2605 ##STR00620## 439
491.3114 ##STR00621## 440 406.21 ##STR00622## 442 ##STR00623## 443
##STR00624## 444 422.2306 ##STR00625## 445 522.283 ##STR00626## 446
471 ##STR00627## 447 387.04 ##STR00628## 448 521.298 ##STR00629##
449 ##STR00630## 450 578.3204 ##STR00631## 451 549.3301
##STR00632## 452 ##STR00633## 453 ##STR00634## 454 ##STR00635## 455
##STR00636## 456 ##STR00637## 457 ##STR00638## 458 ##STR00639## 459
##STR00640## 460 ##STR00641## 461 ##STR00642## 462 491.3124
##STR00643## 463 534.32 ##STR00644## 464 548.34 ##STR00645## 465
550.3506 ##STR00646## 466 323.19 ##STR00647## 467 448.28
##STR00648## 468 548.34 ##STR00649## 469 541.2626 ##STR00650## 434
294.1718 ##STR00651## 435 293.1757 ##STR00652## 441
Biological Activity
[0474] Binding of cytokines and certain growth factors to their
respective receptors trigger activation of Janus kinases, which
phosphorylate members of the STAT family. Phosphorylated STAT
molecules dimerize and migrate to the nucleus where they bind to
DNA and initiate transcription of responsive genes. Inhibitors of
pathways downstream of cytokine/growth factor receptors have
therapeutic potential for several indications. An enzymatic assay
for JAK-3 and JAK-2 has been developed to identify T-cell selective
inhibitors. GST fusion constructs of the kinase domains of both
enzymes are used and a biotinylated, tyrosine containing peptide
serves as substrate. Phosphorylation of this peptide by the
respective kinase is quantified with an europium-labeled
anti-phosphotyrosine antibody (Eu-PT66) as energy donor and a
streptavidin-allophycocyanine conjugate (SA-APC) as energy
acceptor. The assay has been established in a 384 well format.
[0475] In the JAK LANCE.TM. assay, a biotinylated peptide is
incubated together with compounds and ATP in buffer. The
phosphorylation reaction starts after addition of JAK kinase. After
incubation at RT the reaction is stopped with EDTA and the product
detected by addition Streptavidin-Allophycocyanin and
Europium-labeled antiphosphotyrosine antibody. The signal is
measured using an EnVision Reader. Exc: 320 nm, Em, Donor: 615 nm
and Em, Acceptor: 665 nm in time resolved fashion with a delay of
60 s and a window of 100 s.
[0476] Data acquired for the compounds of the invention using these
assays are shown in Tables A and B.
[0477] In order to test the CDK4 activity of the compounds of the
invention, an ELISA based assay can be utilized, where the enzyme
is a purified active CDK4/Cyclin-D1 kinase complex and the
substrate is a purified Retinoblastoma (Rb) protein. The active
CDK4/Cyclin-D1 kinase complex phosphorylates the Rb substrate at
Serine-780 residue, and then the phosphorylated Rb/S780 is detected
via an antibody specific to the phosphorylated site. The compounds
that inhibit the CDK4 kinase activity would inhibit the signal
output of the assay. Data acquired for the compounds of the
invention using this assay are shown in Table C.
[0478] In order to test the CDK2 activity of the compounds of the
invention, the CDK2 assay is a flourescence polarization assay,
where the enzyme is a purified active CDK2/Cyclin-A kinase complex
and the substrate is a synthesized peptide derived from Histone H1.
This assay utilizes the IMAP technology from Molecular Devices. The
active CDK2/Cyclin-A complex phosphorylates the peptide substrate,
which is conjugated with the TAMRA tag. The phosphorylated site is
then recognized by a metal containing molecule that interacts with
the TAMRA tag to induce a high flourescence polarization. The
compounds that inhibit the CDK2 kinase activity would inhibit the
flourescence output of the assay. Data acquired for the compounds
of the invention using this assay are shown in Table C.
p-pRb/S780 ELISA Cellular Assay
[0479] Maxisorp plates (Nunc 442404) are coated with 50 ul of 1
ug/mL total phospholated Retinoblast Protein (pRb) antibody (4H1
Cell Signaling 9309L) diluted in DPBS (Phosphate Buffered Saline)
overnight at 4.degree. C. The next day plates are blocked with
Superblock in TBST (Pierce 37535) for one hour to
overnight--changing block once during that time. Cells are plated
at 50-60% confluency in a 96 well plate (Corning 3585) in 100 uL
complete media (media containing fetal bovine serum (Gibco
1600-044), 2 mM L-Glutamine (Gibco 25030), and 1%
Penicillin/Streptomycin (Gibco 15140-122) and grown overnight in a
humidified chamber at 37.degree. C. and 5% CO.sub.2. Compounds (in
DMSO) are diluted in media to create a 7 point dilution series of
compound with concentrations ranging from 110 uM to 0.027 uM. 10 ul
of the diluted compounds are added to the cells, with final
concentrations on cells ranging from 10 uM to 0.002 uM. Cells are
treated for 24 hrs in a humidified chamber at 37.degree. C. and 5%
CO.sub.2. Following compound incubation, cells are lysed with 40
uL/well lysis buffer (50 mM Tris-HCL pH 7.5 (Invitrogen 15567-027),
120 mM NaCl (Promega V4221), 1 mM EDTA (Gibco 15575-038), 6 mM EGTA
(Fisher 02783-100), 1% Nonidet P40 (Fluka R02771). Plates are
placed on Titerplate shaker (Labline model 4625) for 5 minutes at
4.degree. C. to lyse cells. After lysis, 10 ul of cell lysate and
50 ul xPBS/10% Superblock (Gibco 10010 and Pierce 37535) is added
to each well of the precoated and blocked Maxisorp plate and
allowed to bind at room temperature for 2 hours on Oribtron Rotator
II (Boekel Industries Model 260250). Plates are then washed
3.times. with 1.times.TBST (Teknova T9201) using Biotek platewasher
equipped with a Biostack. The final wash is not aspirated. The
final wash is removed by flicking off and tapping plate on paper
towels. ppRbS780 antibody (Cell Signaling 9307L) is diluted 1:1000
in 1.times.PBS/10% Superblock (Gibco 10010 and Pierce 37535) and 50
ul is added to each well. Plates are then incubated 1 hour on
Oribtron Rotator II (Boekel Industries Model 260250). Plates are
then washed as previously described. Goat anti-rabbit HRP (Promega
W401B) is diluted 1:2500 1.times.PBS/10% Superblock (Gibco 10010
and Pierce 37535) and 50 ul is added to each well. Plates are then
incubated 30 minutes on Oribtron Rotator II. Plates are then washed
as previously described. 50 uL Ultra TMB ELISA (Pierce 34028) is
then added to each well. Plates are incubated 5-20 minutes until
blue color develops. 50 ul 2M Sulfuric acid (Mallinckrodt 2468-46)
is then added to each well to stop the reaction. Absorbance at 450
nm for each plate is read on Spectramax Plus (Molecular Devices).
The results of this assay are summarized in Table E.
BrdU Assay
[0480] Cell Proliferation ELISA BrdU (colorimetric) kit from Roche
Diagnostic (Cat. #: 11647229001, 9115 Hague Road, Indianapolis,
Ind. 50414) is used for this assay. Briefly, cells are plated in 96
well plates at 50-60% confluency in RMPI 1640 media. The next day,
cells are treated with compounds at a desired concentration range
and then incubated for 24 hrs in a humidified chamber at 37.degree.
C. and 5% CO.sub.2. Following the protocol provided by the kit,
cells are labeled with BrdU labeling agent for 2 hrs, and then
fixed with 200 uL of FixDenat for 30 min at room temperature. 100
uL of anti-BrdU antibody is added to the cells and incubated for 2
hrs at room temperature. The cells are then washed three times with
200 uL/well of PBS, and then 100 uL of color developing solution is
added per well. After 5-10 min incubation, the absorbance is read
at 370 nM using Spectramax Plus (Molecular Devices). The results of
this assay are summarized in Table E.
TABLE-US-00026 TABLE E CDK4 ELISA assay CDK4 CDK2cyA hCDK1/ Example
IC50 HTRF/IC50 IMAP/IC50 B/IC50 Number [umol l-] [umol l - 1] [umol
l - 1] [umol l - 1] 201 <10 201A <10 <1 205 <10 <10
206 >10 >10 207 <10 >10 208 <10 <10 209 <10
<10 210 >10 211 <10 <10 212 <10 <1 213 <10
<1 214 <10 >10 215 >10 216 <10 <10 265 <10
<10 266 <1 <10 <10 266A <1 <1 <10 <10 267
<10 <10 >10 >10 217 >10 218 >10 263 >10 >10
269 <10 >10 264 >10 >10 423 >10 424 >10 <1
>10 252 >10 >10 253 <10 <10 254 >10 <1 330
>10 <1 255 >10 <10 >10 269 >10 319 >10 360
>10 >10 361 <1 <1 <1 362 >10 <1 274 >10 270
>10 271 <10 >10 331 >10 <1 329 >10 <1 426
<10 <1 >10 275 8 >10 >10 379 >10 <10 >10
380 >10 <10 381 >10 >10 382 >10 <1 <10 427
>10 >10 364 >10 <1 <10 256 <10 <1 <10 276
<10 >10 >10 373 <10 <10 <10 280 <10 >10
>10 375 <1 <1 <1 374 <1 <1 <1 428 >10 383
<1 <1 <1 429 >10 257 <10 <10 387 <1 <1 10
430 <1 <1 <1 326 >10 10 333 <10 10 10 219 <10
<10 <10 220 >10 >10 221 <10 <10 <10 222 <10
<10 <10 223 <10 <10 >10 224 >10 >10 225 <1
<1 <10 226 <10 <10 >10 227 <10 <10 228 <10
<10 229 <10 230 <1 <10 231 <1 >10 232 <1 <1
376 <1 <10 377 <1 <1 398 <1 <1 <1 234 <10
>10 235 <10 >10 399 <1 <10 399A <1 <10 <10
399C <1 >15 <10 432 >10 >10 433 <1 >10 283
<10 >10 285 <10 <10 258 >10 <10 202 <1 <10
434 >10 >10 435 >10 <10 261 >10 <10 262 >10
<10 385 >10 >10 408 >10 >10 273 >10 >10 437
<10 >10 236 <10 <1 237 <10 <10 438 <10 <10
238 <1 <10 <10 384 <1 >10 239 <1 <10 <1 440
<10 >10 320 <10 >10 240 <1 <10 241 <10 <10
242 <1 <10 388 <10 >10 246 <1 <1 321 <10
>10 287 10 >10 404 <1 >10 405 <10 >10 243 <1
<1 244 <1 <1 245 <1 <1 378 <1 <10 441 10
>10 336 <10 >10 337 <10 >10 442 10 <10 443 10
>10 363 <1 <1 247 <1 <1 <1 335 <1 <10 343
<10 >10 344 <10 >10 444 <1 >10 446 10 <10 286
<10 <10 447 10 >10 345 <1 <10 345A <1 <10
<1 288 <10 <1 322 <10 >10 293 10 >10 334 <1
>10 248 <1 <10 249 <10 <10 366 <10 <10 367
<10 <10 294 10 <10 295 10 >10 296 10 >10 297 10
>10 299 10 <10 300 <10 <1 301 10 >10 298 <10
<1 346 <1 <10 <10 250 <1 <1 <1 303 >10
>10 >10 304 >10 >10 >10 391 <1 >10 >10 305
>10 <10 <10 406 <1 >10 >10 368 <1 <1 <10
449 <10 <10 <10 369 <10 <10 <10 370 <10 <10
<10 371 <10 <10 <1 372 <10 <1 <10 232 >10
>10 >10 306 >10 >10 >10 324 <1 >10 >10 325
<10 >10 >10 389 <1 <10 >10 400 <1 <1 <1
386 <1 <10 <10 386A <1 >10 <10 277 >10 >10
>10 278 <10 >10 >10 312 <10 <10 <10 392 <10
<10 >10 279 <1 <10 <10 393 <1 <10 <10 407
<1 <10 <10 302 <10 >10 >10 457 >10 >10
>10 311 <10 >10 >10 313 <1 <10 <10 347 <1
>10 >10 348 <1 >15 >15 349 <10 >15 >15 350
<1 >15 >10 782 <1 >15 <10 351 <1 <10 <10
352 <1 >10 <10 353 <1 >15 >15 282 >10 >15
>15 284 >10 >15 >15 462 <10 >15 >15 354 <10
>15 >15 314 <1 <10 <10 356 <1 <10 <10 357
<1 <10 <10 358 <1 >15 >15 359 <1 <10 <10
397 <1 >15 >15 281 <10 <10 <10 401 <1 <1
<1 402 <1 <1 <1 315 <1 <1 <10 315A <1
<10 <10 316 <1 <1 <10 463 <1 <10 <10 338
<1 <10 <10 339 <1 >10 >10 340 <1 <10 <10
290 <1 <10 <10 465 <1 <10 <10 291 <1 >10
<10 341 <1 >10 <10 342 <1 <10 <10 292 <10
<10 <10 403 <1 <10 <10 466 <10 <10 <10 467
<10 >15 >15 468 >10 >15 >15 394 <1 >10
>15 395 <1 <1 <1 396 <1 <1 <1
EQUIVALENTS
[0481] Those skilled in the art will recognize, or be able to
ascertain using no more than routine experimentation, many
equivalents to the specific embodiments and methods described
herein. Such equivalents are intended to be encompassed by the
scope of the following claims.
INCORPORATION BY REFERENCE
[0482] The entire contents of all patents, published patent
applications and other references cited herein are hereby expressly
incorporated herein in their entireties by reference.
* * * * *
References