U.S. patent application number 12/191613 was filed with the patent office on 2008-12-04 for substituted thiatriazaacenaphthylene-6-carbonitrile kinase inhibitors.
Invention is credited to Peter J. Connolly, Stuart L. Emanuel, Stuart Hayden, Sigmond G. Johnson, Bharat Lagu, Steven A. Middleton, Niranjan B. Pandey, Mark T. Powell.
Application Number | 20080300264 12/191613 |
Document ID | / |
Family ID | 38345934 |
Filed Date | 2008-12-04 |
United States Patent
Application |
20080300264 |
Kind Code |
A1 |
Connolly; Peter J. ; et
al. |
December 4, 2008 |
SUBSTITUTED THIATRIAZAACENAPHTHYLENE-6-CARBONITRILE KINASE
INHIBITORS
Abstract
The present invention is directed to substituted
thiatriazaacenaphthylene-6-carbonitrile compounds of formula (I):
##STR00001## and forms thereof, their synthesis and use for
treating, preventing or ameliorating a chronic or acute protein
kinase mediated disease, disorder or condition.
Inventors: |
Connolly; Peter J.; (New
Providence, NJ) ; Emanuel; Stuart L.; (Doylestown,
PA) ; Hayden; Stuart; (Point Pleasant, NJ) ;
Johnson; Sigmond G.; (Flemington, NJ) ; Lagu;
Bharat; (Hillsborough, NJ) ; Middleton; Steven
A.; (Flemington, NJ) ; Pandey; Niranjan B.;
(White Marsh, MD) ; Powell; Mark T.; (Newtown,
PA) |
Correspondence
Address: |
PHILIP S. JOHNSON;JOHNSON & JOHNSON
ONE JOHNSON & JOHNSON PLAZA
NEW BRUNSWICK
NJ
08933-7003
US
|
Family ID: |
38345934 |
Appl. No.: |
12/191613 |
Filed: |
August 14, 2008 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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11672177 |
Feb 7, 2007 |
7427625 |
|
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12191613 |
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60771256 |
Feb 8, 2006 |
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Current U.S.
Class: |
514/267 ;
544/250 |
Current CPC
Class: |
C07D 495/06
20130101 |
Class at
Publication: |
514/267 ;
544/250 |
International
Class: |
A61K 31/519 20060101
A61K031/519; C07D 495/16 20060101 C07D495/16 |
Claims
1.-19. (canceled)
20. A process for preparing a pharmaceutical composition comprising
the step of admixing a pharmaceutically acceptable carrier and a
compound of Formula 1: ##STR00041## and a form thereof, wherein
R.sub.1 is selected from the group consisting of --N(Ra,Rb),
--O(Ra) and --N(Ra)-(CH.sub.2).sub.p--Ar.sup.1; Ra is selected from
the group consisting of hydrogen and C.sub.1-8alkyl; Rb is selected
from the group consisting of hydrogen, C.sub.1-8alkyl and
hydroxy-(C.sub.1-8)alkyl; alternatively, when R.sub.1 is
--N(Ra,Rb), then Ra and Rb may be taken together with the nitrogen
of attachment to form a heterocyclyl ring having at least one said
nitrogen atom, wherein said ring is optionally substituted with
one, two or three substituents each selected from the group
consisting of C.sub.1-8alkyl, C.sub.2-8alkenyl, C.sub.2-8alkynyl,
C.sub.1-8-alkoxy, halogen, hydroxy, hydroxy-(C.sub.1-8)alkyl,
hydroxy-(C.sub.1-8)alkoxy, amino and C.sub.1-8alkyl-amino; p is 0,
1, 2, 3 or 4; Ar.sup.1 is selected from the group consisting of
aryl, heteroaryl, benzofused heteroaryl, heterocyclyl and
benzofused heterocyclyl each optionally substituted with one, two
or three substituents each selected from the group consisting of
R.sub.4--(C.sub.1-8)alkyl, R.sub.4--(C.sub.2-8)alkenyl,
R.sub.4--(C.sub.2-8)alkynyl, R.sub.4--(C.sub.1-8)alkoxy, cyano,
halogen, nitro, halo-(C.sub.1-8)alkyl, halo-(C.sub.1-8)alkoxy,
hydroxy, hydroxy-(C.sub.1-8)alkyl, hydroxy-(C.sub.1-8)alkoxy,
C.sub.1-8alkoxy-carbonyl, C.sub.1-8alkoxy-carbonyl-amino,
amino-(C.sub.1-8)alkyl, C.sub.1-8alkyl-amino-(C.sub.1-8)alkyl,
amino, C.sub.1-8alkyl-amino, C.sub.3-8cycloalkyl-amino,
heterocyclyl-amino (optionally substituted on heterocyclyl with one
or two C.sub.1-8alkyl substituents) and
heterocyclyl-(C.sub.1-8)alkyl-amino (optionally substituted on
heterocyclyl with one or two C.sub.1-8alkyl substituents); R.sub.4
is hydrogen or is one, two or three substituents each selected from
the group consisting of C.sub.3-8cycloalkyl, aryl, heteroaryl and
heterocyclyl each optionally substituted with one, two or three
substituents each selected from the group consisting of
C.sub.1-8alkyl, C.sub.1-8alkoxy, C.sub.1-8alkoxy-(C.sub.1-8)alkyl,
cyano, halogen, hydroxy, halo-(C.sub.1-8)alkyl,
halo-(C.sub.1-8)alkoxy, hydroxy-(C.sub.1-8)alkyl,
hydroxy-(C.sub.1-8)alkoxy, C.sub.1-8alkoxy-carbonyl,
amino-(C.sub.1-8)alkyl, C.sub.1-8alkyl-amino-(C.sub.1-8)alkyl,
amino, C.sub.1-8alkyl-amino, C.sub.1-8alkoxy-carbonyl-amino;
R.sub.2 is selected from the group consisting of aryl, heteroaryl,
benzofused heteroaryl, heterocyclyl and benzofused heterocyclyl
each optionally substituted with one, two or three substituents
each selected from the group consisting of C.sub.1-8alkyl,
C.sub.2-8alkenyl, C.sub.2-8alkynyl, C.sub.1-8alkoxy, cyano,
halogen, halo-(C.sub.1-8)alkyl, halo-(C.sub.1-8)alkoxy, hydroxy,
heteroaryl-oxy, aryl-oxy (optionally substituted on heteroaryl and
aryl with one, two or three substituents each selected from the
group consisting of C.sub.1-8alkyl, C.sub.1-8alkoxy and halogen);
and R.sub.3 is hydrogen or is selected from the group consisting of
R.sub.4--(C.sub.1-8)alkyl, R.sub.4--(C.sub.2-8)alkenyl,
R.sub.4--(C.sub.2-8)alkynyl, halogen, hydroxy, C.sub.3-cycloalkyl,
heteroaryl, aryl and heterocyclyl, optionally substituted on
heteroaryl and aryl with one, two or three substituents each
selected from the group consisting of C.sub.1-8alkyl,
C.sub.1-8alkoxy and halogen.
21.-29. (canceled)
30. A process for preparing a compound of Formula 1 comprising the
steps of: a. reacting a compound of Formula A1 with a compound of
Formula A2 in the presence of a base to provide a compound of
Formula A3: ##STR00042## b. reacting the compound of Formula A3
with a compound of Formula A4 (wherein Ra is as defined herein) in
the presence of a weak organic base to provide a compound of
Formula A5: ##STR00043## c. reacting the compound of Formula A5
with a compound of Formula A6 in the presence of a non-nucleophilic
base to provide a compound of Formula A7: ##STR00044## d. reacting
the compound of Formula A7 with a carbonyl source in the presence
of a non-nucleophilic base to provide a compound of Formula A8,
representative of a compound of any of claim 1 to 27, wherein
R.sub.1 is --O(Ra): ##STR00045## e. reacting the compound of
Formula A8 with a halogenating reagent to provide a compound of
Formula A9 (wherein X is a halogen atom), representative of a
compound of any of claim 1 to 27, wherein R.sub.1 is --O(Ra):
##STR00046## f. reacting the compound of Formula A9 under catalytic
conditions with a compound of Formula A10 (wherein Y and R.sub.3
are both hydrogen) in the optional presence of a base, to provide a
compound of Formula A11, representative of a compound of any of
claim 1 to 27, wherein R.sub.1 is --O(Ra): ##STR00047## wherein
said compound of Formula 1 is ##STR00048## and a form thereof,
wherein R.sub.1 is selected from the group consisting of
--N(Ra,Rb), --O(Ra) and --N(Ra)-(CH.sub.2).sub.p--Ar.sup.1; Ra is
selected from the group consisting of hydrogen and C.sub.1-8alkyl;
Rb is selected from the group consisting of hydrogen,
C.sub.1-8alkyl and hydroxy-(C.sub.1-8)alkyl; alternatively, when
R.sub.1 is --N(Ra,Rb), then Ra and Rb may be taken together with
the nitrogen of attachment to form a heterocyclyl ring having at
least one said nitrogen atom, wherein said ring is optionally
substituted with one, two or three substituents each selected from
the group consisting of C.sub.1-8alkyl, C.sub.2-8alkenyl,
C.sub.2-8alkynyl, C.sub.1-8alkoxy, halogen, hydroxy,
hydroxy-(C.sub.1-8allyl, hydroxy-(C.sub.1-8)alkoxy, amino and
C.sub.1-8alkyl-amino; p is 0, 1, 2, 3 or 4; Ar.sup.1 is selected
from the group consisting of aryl, heteroaryl, benzofused
heteroaryl, heterocyclyl and benzofused heterocyclyl each
optionally substituted with one, two or three substituents each
selected from the group consisting of R.sub.4--(C.sub.1-8)alkyl,
R.sub.4--(C.sub.2-8)alkenyl, R.sub.4--(C.sub.2-8)alkynyl,
R.sub.4--(C.sub.1-8)alkoxy, cyano, halogen, nitro,
halo-(C.sub.1-8)alkyl, halo-(C.sub.1-8)alkoxy, hydroxy,
hydroxy-(C.sub.1-8)alkyl, hydroxy-(C.sub.1-8)alkoxy,
C.sub.1-8alkoxy-carbonyl, C.sub.1-8alkoxy-carbonyl-amino,
amino-(C.sub.1-8)alkyl, C.sub.1-8alkyl-amino-(C.sub.1-8)alkyl,
amino, C.sub.1-8alkyl-amino, C.sub.3-8cycloalkyl-amino,
heterocyclyl-amino (optionally substituted on heterocyclyl with one
or two C.sub.1-8alkyl substituents) and
heterocycyl-(C.sub.1-8)alkyl-amino (optionally substituted on
heterocyclyl with one or two C.sub.1-8alkyl substituents); R.sub.4
is hydrogen or is one, two or three substituents each selected from
the group consisting of C.sub.3-8cycloalkyl, aryl, heteroaryl and
heterocyclyl each optionally substituted with one, two or three
substituents each selected from the group consisting of
C.sub.1-8alkyl, C.sub.1-8alkoxy, C.sub.1-8alkoxy-(C.sub.1-8)alkyl,
cyano, halogen, hydroxy, halo-(C.sub.1-8)alkyl,
halo-(C.sub.1-8)alkoxy, hydroxy-(C.sub.1-8)alkyl,
hydroxy-(C.sub.1-8)alkoxy, C.sub.1-8alkoxy-carbonyl,
amino-(C.sub.1-8)alkyl, C.sub.1-8alkyl-amino-(C.sub.1-8)alkyl,
amino, C.sub.1-8alkyl-amino, C.sub.1-8alkoxy-carbonyl-amino;
R.sub.2 is selected from the group consisting of aryl, heteroaryl,
benzofused heteroaryl, heterocyclyl and benzofused heterocyclyl
each optionally substituted with one, two or three substituents
each selected from the group consisting of C.sub.1-8alkyl,
C.sub.2-8alkenyl, C.sub.2-8alkynyl, C.sub.1-8alkoxy, cyano,
halogen, halo-(C.sub.1-8)alkyl, halo-(C.sub.1-8)alkoxy, hydroxy,
heteroaryl-oxy, aryl-oxy (optionally substituted on heteroaryl and
aryl with one, two or three substituents each selected from the
group consisting of C.sub.1-8alkyl, C.sub.1-8alkoxy and halogen);
and R.sub.3 is hydrogen or is selected from the group consisting of
R.sub.4--(C.sub.1-8)alkyl, R.sub.4--(C.sub.2-8)alkenyl,
R.sub.4--(C.sub.2-8)alkynyl, halogen, hydroxy, C.sub.3-cycloalkyl,
heteroaryl, aryl and heterocyclyl, optionally substituted on
heteroaryl and aryl with one, two or three substituents each
selected from the group consisting of C.sub.1-8alkyl,
C.sub.1-8alkoxy and halogen.
31. A process for preparing a compound of Formula 1 comprising the
steps of: a. reacting a compound of Formula B1 (representative of a
compound of any of claim 1 to 27, wherein R.sub.1 is --O(Ra) and Ra
is alkyl) with a basic solution to provide a compound of Formula
B2, representative of a compound of any of claim 1 to 27, wherein
R.sub.1 is --O(Ra) and Ra is hydrogen: ##STR00049## b. reacting the
compound of Formula B2 with a compound of Formula B3 (wherein Rx
represents NH(Ra,Rb), NH(Ra,Rb) wherein Ra and Rb are taken
together with the nitrogen of attachment to form a heterocyclyl
ring and NH(Ra)-(CH.sub.2).sub.p--Ar.sup.1) to provide a compound
of Formula B4, representative of a compound of any of claim 1 to
27, wherein R.sub.1 is selected from --N(Ra,Rb), --N(Ra,Rb) wherein
Ra and Rb are taken together with the nitrogen of attachment to
form a heterocyclyl ring and --N(Ra)-(CH.sub.2).sub.p--Ar.sup.1:
##STR00050## wherein said compound of Formula 1 is ##STR00051## and
a form thereof, wherein R.sub.1 is selected from the group
consisting of --N(Ra,Rb), --O(Ra) and
--N(Ra)-(CH.sub.2).sub.p--Ar.sup.1; Ra is selected from the group
consisting of hydrogen and C.sub.1-8alkyl; Rb is selected from the
group consisting of hydrogen, C.sub.1-8alkyl and
hydroxy-(C.sub.1-8)alkyl; alternatively, when R.sub.1 is
--N(Ra,Rb), then Ra and Rb may be taken together with the nitrogen
of attachment to form a heterocyclyl ring having at least one said
nitrogen atom, wherein said ring is optionally substituted with
one, two or three substituents each selected from the group
consisting of C.sub.1-8alkyl, C.sub.2-8alkenyl, C.sub.2-8alkynyl,
C.sub.1-8alkoxy, halogen, hydroxy, hydroxy-(C.sub.1-8)alkyl,
hydroxy-(C.sub.1-8)alkoxy, amino and C.sub.1-8alkyl-amino; p is 0,
1, 2, 3 or 4; Ar.sup.1 is selected from the group consisting of
aryl, heteroaryl, benzofused heteroaryl, heterocyclyl and
benzofused heterocyclyl each optionally substituted with one, two
or three substituents each selected from the group consisting of
R.sub.4--(C.sub.1-8)alkyl, R.sub.4--(C.sub.2-8)alkenyl,
R.sub.4--(C.sub.2-8)alkynyl, R.sub.4--(C.sub.1-8)alkoxy, cyano,
halogen, nitro, halo-(C.sub.1-8)alkyl, halo-(C.sub.1-8)alkoxy,
hydroxy, hydroxy-(C.sub.1-8)alkyl, hydroxy-(C.sub.1-8)alkoxy,
C.sub.1-8alkoxy-carbonyl, C.sub.1-8alkoxy-carbonyl-amino,
amino-(C.sub.1-8)alkyl, C.sub.1-8alkyl-amino-(C.sub.1-8)alkyl,
amino, C.sub.1-8alkyl-amino. C.sub.3-8cycloalkyl-amino,
heterocyclyl-amino (optionally substituted on heterocyclyl with one
or two C.sub.1-8alkyl substituents) and
heterocyclyl-(C.sub.1-8)alkyl-amino (optionally substituted on
heterocyclyl with one or two C.sub.1-8alkyl substituents); R.sub.4
is hydrogen or is one, two or three substituents each selected from
the group consisting of C.sub.3-8cycloalkyl, aryl, heteroaryl and
heterocyclyl each optionally substituted with one, two or three
substituents each selected from the group consisting of
C.sub.1-8alkyl, C.sub.1-8alkoxy, C.sub.1-8alkoxy-(C.sub.1-8)alkyl,
cyano, halogen, hydroxy, halo-(C.sub.1-8)alkyl,
halo-(C.sub.1-8)alkoxy, hydroxy-(C.sub.1-8)alkyl,
hydroxy-(C.sub.1-8)alkoxy, C.sub.1-8alkoxy-carbonyl,
amino-(C.sub.1-8)alkyl, C.sub.1-8-alkyl-amino-(C.sub.1-8)alkyl,
amino, C.sub.1-8alkyl-amino, C.sub.1-8alkoxy-carbonyl-amino;
R.sub.2 is selected from the group consisting of aryl, heteroaryl,
benzofused heteroaryl, heterocyclyl and benzofused heterocyclyl
each optionally substituted with one, two or three substituents
each selected from the group consisting of C.sub.1-8alkyl,
C.sub.2-8alkenyl, C.sub.2-8alkynyl, C.sub.1-8alkoxy, cyano,
halogen, halo-(C.sub.1-8)alkyl, halo-(C.sub.1-8)alkoxy, hydroxy,
heteroaryl-oxy, aryl-oxy (optionally substituted on heteroaryl and
aryl with one, two or three substituents each selected from the
group consisting of C.sub.1-8alkyl, C.sub.1-8alkoxy and halogen);
and R.sub.3 is hydrogen or is selected from the group consisting of
R.sub.4--(C.sub.1-8)alkyl, R.sub.4--(C.sub.2-8)alkenyl,
R.sub.4--(C.sub.2-8)alkynyl, halogen, hydroxy, C.sub.3-cycloalkyl,
heteroaryl, aryl and heterocyclyl, optionally substituted on
heteroaryl and aryl with one, two or three substituents each
selected from the group consisting of C.sub.1-8alkyl,
C.sub.1-8alkoxy and halogen.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This present application is a divisional application of U.S.
application Ser. No. 11/672,177 and claims priority of the benefits
benefit of U.S. application Ser. No. 11/672,177 filed Feb. 7, 2007,
and Provisional Patent Application Ser. No. 60/771,256, filed Feb.
8, 2006. The complete disclosures of the aforementioned related US
applications are which is incorporated herein by reference in its
their entirety and for all purposes.
FIELD OF THE INVENTION
[0002] The present invention is in the area of substituted
thiatriazaacenaphthylene-6-carbonitrile compounds or forms thereof,
their syntheses and their use as kinase inhibitors.
BACKGROUND OF THE INVENTION
[0003] In general, protein kinases are the largest set of
structurally related phosphoryl transferases, have highly conserved
structures and catalytic functions and may be categorized into
families by the substrates they phosphorylate (e.g.,
protein-tyrosine, protein-serine/threonine, histidine and the like)
and are responsible for the control of a wide variety of cellular
signal transduction processes.
[0004] Examples of protein-tyrosine kinases include, but are not
limited to, Irk, IGFR-1, Zap-70, Bmx, Btk, CHK (Csk homologous
kinase), CSK (C-terminal Src Kinase), Itk-1, Src (c-Src, Lyn, Fyn,
Lck, Syk, Hck, Yes, Blk, Fgr and Frk), Tec, Txk/Rlk, Abl, EGFR
(EGFR-1/ErbB-1, ErbB-2/NEU/HER-2, ErbB-3 and ErbB-4), FAK, FGF1R
(also FGFR1 or FGR-1), FGF2R (also FGR-2), MET (also Met-1 or
c-MET), PDGFR (.alpha. and .beta.), Tie-1, Tie-2 (also Tek-1 or
Tek), VEGFR1 (also FLT-1), VEGFR2 (also KDR), FLT-3, FLT-4, c-KIT,
JAK1, JAK2, JAK3, TYK2, LOK, RET, TRKA, PYK2, ALK (Anaplastic
Lymphoma Kinase), EPHA (1-8), EPHB (1-6), RON, Fes, Fer or EPHB4
(also EPHB4-1).
[0005] Examples of protein-serine/threonine kinases include, but
are not limited to, Ark, ATM (1-3), CamK (I-IV), CamKK, Chk1 and 2
(Checkpoint kinases), CK1, CK2, Erk, IKK-I (also IKK-ALPHA or
CHUK), IKK-2 (also IKK-BETA), Ilk, Jnk (1-3), LimK (1 and 2),
MLK3Raf (A, B, and C), CDK (1-10), PKC (including all PKC
subtypes), Plk (1-3), NIK, Pak (1-3), PDK1, PKR, RhoK, RIP, RIP-2,
GSK3 (.alpha. and .beta.), PKA, P38, Erk (1-3), PKB (including all
PKB subtypes) (also AKT-1, AKT-2, AKT-3 or AKT3-1), IRAK1, FRK,
SGK, TAK1 or Tpl-2 (also COT).
[0006] Protein kinases play very important roles in the normal
regulation of cell growth. However, as a result of dysregulation of
the tyrosine kinases (receptor or non-receptor) or the ligands of
the receptor tyrosine kinases, signaling can become deregulated,
resulting in uncontrolled cell proliferation leading to cancer or a
related disease, disorder or syndrome.
[0007] Protein kinases catalyze and regulate the process of
phosphorylation, whereby the kinases covalently attach phosphate
groups to proteins or lipid targets in response to a variety of
extracellular signals: hormones, neurotransmitters, growth and
differentiation factors, cell cycle events, environmental stresses,
nutritional stresses and the like.
[0008] Phosphorylation modulates or regulates a variety of cellular
processes such as proliferation, growth, differentiation,
metabolism, apoptosis, motility, transcription, translation and
other signaling processes. Defective control of protein
phosphorylation due to unregulated cellular mitosis, unregulated
cell proliferation and upregulated kinase activity has been
implicated in a number of diseases and disease conditions, such as
osteoarthritis, rheumatoid arthritis, synovial pannus invasion in
arthritis, multiple sclerosis, myasthenia gravis, diabetes
mellitus, diabetic angiopathy, diabetic retinopathy, retinal vessel
proliferation, inflammatory bowel disease, Crohns disease,
ulcerative colitis, bone diseases, transplant or bone marrow
transplant rejection, lupus, chronic pancreatitis, cachexia, septic
shock, fibroproliferative and differentiative skin diseases or
disorders, central nervous system diseases, neurodegenerative
diseases, disorders or conditions related to nerve damage and axon
degeneration subsequent to a brain or spinal cord injury, acute or
chronic cancer, occular diseases, viral infections, heart disease,
lung or pulmonary diseases or kidney or renal diseases. Therefore,
kinase inhibitors have potential use as therapeutic agents.
[0009] The term "myasthenia gravis" means a disease having the
characteristic feature of easy fatigue of certain voluntary muscle
groups on repeated use. Muscles of the face or upper trunk are
especially likely to be affected. In most and perhaps all cases,
the disease is due to the development of autoantibodies against the
acetylcholine receptor in neuromuscular junctions. Immunization of
animals with this receptor protein leads to a disease with the
features of myasthenia gravis.
[0010] In reference to "synovial pannus invasion in arthritis," the
term "pannus" means a disease whereby vascularised granulation
tissue rich in fibroblasts, lymphocytes and macrophages, derived
from synovial tissue, overgrows the bearing surface of the joint in
rheumatoid arthritis and is associated with the breakdown of the
articular surface.
[0011] The tyrosine kinases can further be categorized by whether
they are receptor tyrosine kinases or non-receptor tyrosine
kinases. The receptor tyrosine kinases span the cell membrane with
a ligand interacting domain protruding from the cell, with a
hydrophobic trans-membrane domain, and a cytoplasmic domain that
contains the catalytic kinase domain and other regulatory
sequences. Non-receptor tyrosine kinases are often myristylated or
modified by the addition of other hydrophobic moieties that allow
them to be anchored to the cell membrane.
[0012] The epidermal growth factor receptor (EGFR) tyrosine-kinase
family includes the receptors EGFR (also referred to as EGFR-1 or
Erb-B1), HER-2 (or neu), EGFR3 and EGFR4. Epidermal Growth Factor
(EGF), Transforming Growth Factor-.alpha. (TGF-.alpha.) and the
HER-2 ligand heregulin are three of the ligands that bind to the
EGFR receptors.
[0013] EGFR overexpression or mutation of one or more EGFR kinase
family members has been commonly involved in cancer and other
diseases characterized by uncontrolled or abnormal cell growth.
Deregulation of EGFR has also been associated with epidermoid
tumors, head and neck tumors, breast tumors and tumors involving
other major organs, such as the lungs and gastrointestinal tract.
The clinically prevalent cancers related to EGFR include lung,
gastric and head and neck cancer (Klijn J G, Berns P M, Schmitz P I
and Foekens J A; The clinical significance of epidermal growth
factor receptor (EGF-R) in human breast cancer: a review on 5232
patients, Endocr. Rev., 1992, 13, 3-17; Salomon D and Gullick W;
The erbB family of receptors and their ligands: Multiple targets
for therapy, Signal, 2001, 2, 4-11). Other diseases associated with
increased EGFR expression include proliferative glomerulonephritis,
diabetes-induced renal disease and chronic pancreatitis.
[0014] In treating cancers of the head such as brain cancers and
the like, the ability of small molecule EGFR inhibitors to
penetrate the blood brain barrier could have therapeutic advantages
since EGFR is often overexpressed in primary brain tumors and also
in breast and non-small cell lung carcinomas that frequently
metastasize to the brain (Eckstrand A J, Sugawa N, James C D and
Collins V P; Amplified and rearranged epidermal growth factor
receptor genes in human glioblastomas reveal deletions of sequences
encoding portions of the N- and/or C-terminal tails, Proc. Acad.
Natl. Sci. USA, 1992, 89, 4309-4313; and, Wickstrand C J, Hale L P,
Batra S K, Hill M L, Humphrey P A, Kurpad S N, McLendon R E,
Moscatello D, Pegram C N, Reist C J, Traweek S T, Wong A J,
Zalutsky M R and Bigner, D D; Monoclonal antibodies against
EGFRvIII are tumor specific and react with breast and lung
carcinomas and malignant gliomas, Cancer Res., 1995, 55,
3140-3148).
[0015] EGFR inhibitors tested in neurite outgrowth assays have
activity in promoting neurite outgrowth in both cerebellar granule
cells and dorsal root ganglion neurons, likely by acting directly
on neurons to block neuronal inhibitory responses to myelin
inhibitors, and thus an EGFR inhibitor may have potential use for
promoting axon regeneration after brain and spinal cord injury (V.
Koprivica, et al, EGFR activation mediates inhibition of axon
regeneration by myelin and chondroitin sulfate proteoglycans,
Science, 2005, 310, 106).
[0016] HER1 and HER2 overexpression has been implicated in a
variety of cancers, such as bladder, breast, colorectal,
endometrial, esophageal, gastric (stomach), glioma head and neck,
lung (non-small cell lung cancer), ovarian, pancreatic, renal and
prostate cancer.
[0017] Comparing the overexpression of HER1 and HER2 in tumors,
according to order of prevalence, HER1 overexpression is found in
breast, renal cell, lung, colorectal, head and neck, ovarian,
pancreatic, glioma, bladder, esophageal, gastric, endometrial and
cervical cancer tumors; in contrast, HER2 overexpression is found
in esophageal, head and neck, lung, gastric, renal cell, breast,
bladder, ovarian and colorectal, prostate and endometrial cancer
tumors (Horizons in Cancer Therapeutics: From Bench to Bedside,
Signal Transduction Inhibitors, 2001, 2(2), ISSN 1532-3048).
[0018] While the degree of HER2 overexpression in breast and
ovarian cancer is not as great as in some other cancers, HER2 has
been found to be responsible for these clinically prevalent cancers
(Slamon D J, Clark G M, Wong S G, Levin W J, Ullrich A and McGuire
W L; Human breast cancer: Correlation of relapse and survival with
amplification of HER-2/neu oncogene, Science, 1987, 235, 177-82;
Slamon D J, Godolphin W, Jones L A, Holt J A, Wong S G, Keith D E,
et al; Studies of the HER-2/neu proto-oncogene in human breast and
ovarian cancer, Science, 1989, 244, 707-712; Hetzel D J, Wilson T
O, Keeney G L, Roche P C, Cha S S and Podrantz K C; HER-2/neu
expression: A major prognostic factor in endometrial cancer,
Gynecol. Oncol., 1992, 47, 179-85).
[0019] Furthermore, patients with HER-2 overexpressing breast
cancer frequently experience metastases to the brain (Kirsch D G
and Hochberg F H; Targeting HER-2 in brain metastases from breast
cancer, Clin. Can. Res., 2003, 9, 5435-5436). These patients have
an extremely poor prognosis and intracerebral tumors are often the
cause of death. Autopsy revealed that 20-30% of patients who die of
breast cancer have brain metastases (Grossi P M, Ochiai H, Archer G
E, McLendon R E, Zalutsky M R, Friedman A H, Friedman H S, Bigner D
D and Sampson J H; Efficacy of intracerebral microinfusion of
trastuzumab in an athymic rat model of intracerebral metastatic
breast cancer, Clin. Can. Res., 2003, 9, 5514-5520).
[0020] Human cytomegalovirus (CMV) is a widespread opportunistic
human herpes virus that causes severe and fatal diseases in those
who are immune compromised and in transplant recipients. CMV is
also a leading cause of atherosclerosis and virally mediated birth
defects. The human CMV uses the EGFR receptor to enter cells during
infection, EGFR is autophosphorylated and the downstream signal
transduction pathway components are activated; however, the EGFR
specific inhibitor tyrphostin AG1478 has been shown to reduce the
viral load in cells that were infected in the presence of the
tyrphostin (Wang X, et al., Nature, 24 Jul. 2003, Vol 424,
456-461). Accordingly, potent EGFR selective inhibitors may be
useful in anti-CMV therapy.
[0021] The Src family of tyrosine-kinases includes the sub-family
proteins c-Src, Lyn, Fyn, Lck, Syk, Hck, Yes, Blk, Fgr and Frk.
While various members of the c-Src family are important for normal
cellular proliferation, their overexpression and overactivation can
promote development of cancer (Yeatman T J, Nature, June 2004, Vol.
4). For example, the Lyn kinase has been shown to be upregulated in
hormone resistant prostate cancer. Tumor xenografts of hormone
resistant prostate cancer cells showed delayed growth upon
treatment with peptides that specifically block Lyn kinase activity
(Goldenberg-Furmanov, et al., Cancer Research, 1 Feb. 2004, 64,
1058-1064).
[0022] The Lyn and Hck Src sub-family tyrosine-kinases have both
been implicated in chronic myeloid leukemia (CML). CML is caused by
the BCR-Abl fusion protein resulting from the t(9;22) chromosomal
translocation that juxtaposes the c-Abl non-receptor tyrosine
kinase gene on chromosome 9 with a breakpoint cluster region (bcr)
gene on chromosome 22. The BCR-Abl fusion protein is a
constitutively activated form of the Abl tyrosine kinase that
drives uncontrolled growth leading to CML and many cases of adult
acute lymphoblastic leukemia. Gleevec is an inhibitor of Abl that
has been successfully used to treat CML. However, Gleevec does not
help patients in blast crisis because they carry mutant forms of
BCR-Abl that no longer bind Gleevec. Such Gleevec resistant CML
cells are sensitive to a dual src/BCR-Abl inhibitor that binds and
inhibits the mutant BCR-Abl and members of the src family (Shah, et
al., Science, 16 Jul. 2004, Vol 305, 399-401). CML cells can also
become resistant to treatment with the tyrosine kinase Abl
inhibitor Gleevec in other ways. For example, CML K562 cells that
become resistant to Gleevec minimize reliance on the BCR-Abl
translocation for growth and instead upregulate the Lyn and Hck
kinases, as demonstrated by expressing antisense Lyn in these
cells, which reduced their rate of proliferation (Donato, et al.,
Blood, 15 Jan. 2003, 101(2)). c-Src and other Src family members
are also involved in cellular adhesion, invasion and motility of
tumor cells. Thus, small molecule inhibitors of the Src kinase
family could offer new therapeutic opportunities for both leukemias
and solid tumors.
[0023] Aurora kinases (Aurora-A, Aurora-B and Aurora-C) are highly
conserved tyrosine kinases found in all organisms where they
function to regulate microtubule dynamics during the M phase of the
cell cycle and are essential for mitotic progression. Aurora-A
kinase associates with the centrosome around the pericentriolar
material, as well as the microtubules at the bipolar
mitotic-spindle poles and the midbody microtubules and plays a role
in spindle formation and organization of the centrosome. Aurora-B
regulates chromosomal movement and cytokinesis and Aurora-C's
biological function is not yet understood. The Aurora-A kinase is
involved in centrosome separation, duplication and maturation as
well as in bipolar spindle assembly and stability. Aurora-A is
overexpressed in a number of different human cancers and tumor cell
lines. Overexpression of Aurora is sufficient to induce growth in
soft agar and transforms cells making them tumorigenic. Inhibition
of Aurora activity results in centrosome/chromosome segregation
defects leading to monopolar spindles and polyploidy which induces
cell apoptosis in a variety of cancer cell lines and has suppressed
tumor growth in vivo.
[0024] Angiogenesis plays a role in various processes including
development of the vasculature, wound healing and maintenance of
the female reproductive system. Pathological angiogenesis is
associated with disease states such as cancer, diabetic
retinopathy, rheumatoid arthritis, endometriosis and psoriasis.
Solid-tumor cancers, in particular, are dependent on angiogenesis
for their growth. The vascular endothelial growth factors (VEGFs)
are mediators of both normal and pathologic angiogenesis. VEGF
transmits signals into cells through their cognate receptors, which
belong to the receptor tyrosine kinase (RTK) family of
transmembrane receptors. These receptors are tripartite, consisting
of an extracellular ligand-binding domain, a transmembrane domain,
which anchors the receptor in the membrane of the cell, and an
intracellular tyrosine kinase domain.
[0025] One subfamily of RTKs comprises the receptors Flt1/VEGF-R1
and KDR/Flk1/VEGF-R2, which bind VEGFs. Binding of the VEGF ligand
to the receptor results in stimulation of the receptor tyrosine
kinase activity and transduction of biological signals into the
cell. The KDR/Flk1/VEGF-R2 receptor mediates the biological
activities of mitogenesis and proliferation of endothelial cells
while the Flt1/VEGF-R1 receptor mediates functions such as
endothelial cell adhesion. Inhibition of KDR/Flk1/VEGF-R2
signalling has been shown to inhibit the process of angiogenesis.
Inhibitors of this receptor are likely useful in controlling or
limiting angiogenesis.
[0026] There is a need for potent small-molecule kinase inhibitors
of one or more of the EGFR, HER-2, c-Src, Lyn, c-Abl, Aurora-A or
VEGF kinase proteins and the like possessing anti-tumor cell
proliferation activity, and as such are useful in treating or
ameliorating a EGFR, HER-2, c-Src, Lyn, c-Abl, Aurora-A or VEGF
kinase receptor mediated, angiogenesis-mediated or
hyperproliferative disorder.
SUMMARY OF THE INVENTION
[0027] The present invention is directed to a compound of Formula
(I):
##STR00002##
and forms thereof, wherein R.sub.1, R.sub.2 and R.sub.3 are as
defined herein.
[0028] An example of the present invention includes using a
compound of formula (I) as a protein kinase inhibitor.
[0029] An example of the present invention includes a method for
using a compound of formula (I) as an inhibitor of a protein kinase
such as EGFR, HER-2, c-Src, Lyn, c-Abl, Aurora-A or VEGF comprising
contacting the protein kinase domain or receptor with the
compound.
[0030] An example of the present invention includes a method for
using a compound of formula (I) and forms, pharmaceutical
compositions or medicaments thereof in treating, preventing or
ameliorating a kinase mediated disorder.
[0031] The present invention is further direct to a method for
treating, preventing or ameliorating a chronic or acute protein
kinase mediated disease, disorder or condition in a subject in need
thereof comprising administering to the subject an effective amount
of a compound of formula (I) or a form thereof.
[0032] These and other aspects and advantages of the invention,
which will become apparent in light of the detailed description
below, are achieved through use of the compounds of this
invention.
DETAILED DESCRIPTION OF THE INVENTION
[0033] The present invention provides
thiatriazaacenaphthylene-6-carbonitrile compounds of Formula
(I):
##STR00003##
and a form thereof, wherein [0034] R.sub.1 is selected from the
group consisting of --N(Ra,Rb), --O(Ra) and
--N(Ra)-(CH.sub.2).sub.p--Ar.sup.1; [0035] Ra is selected from the
group consisting of hydrogen and C.sub.1-8alkyl; [0036] Rb is
selected from the group consisting of hydrogen, C.sub.1-8alkyl and
hydroxy-(C.sub.1-8)alkyl; [0037] alternatively, when R.sub.1 is
--N(Ra,Rb), then Ra and Rb may be taken together with the nitrogen
of attachment to form a heterocyclyl ring having at least one said
nitrogen atom, wherein said ring is optionally substituted with
one, two or three substituents each selected from the group
consisting of C.sub.1-8alkyl, C.sub.2-8alkenyl, C.sub.2-8alkynyl,
C.sub.1-8alkoxy, halogen, hydroxy, hydroxy-(C.sub.1-8)alkyl,
hydroxy-(C.sub.1-8)alkoxy, amino and C.sub.1-8alkyl-amino; [0038] p
is 0, 1, 2, 3 or 4; [0039] Ar.sup.1 is selected from the group
consisting of aryl, heteroaryl, benzofused heteroaryl, heterocyclyl
and benzofused heterocyclyl each optionally substituted with one,
two or three substituents each selected from the group consisting
of R.sub.4--(C.sub.1-8)alkyl, R.sub.4--(C.sub.2-8)alkenyl,
R.sub.4--(C.sub.2-8)alkynyl, R.sub.4--(C.sub.1-8)alkoxy, cyano,
halogen, nitro, halo-(C.sub.1-8)alkyl, halo-(C.sub.1-8)alkoxy,
hydroxy, hydroxy-(C.sub.1-8)alkyl, hydroxy-(C.sub.1-8)alkoxy,
C.sub.1-8alkoxy-carbonyl, C.sub.1-8alkoxy-carbonyl-amino,
amino-(C.sub.1-8)alkyl, C.sub.1-8alkyl-amino-(C.sub.1-8)alkyl,
amino, C.sub.1-8alkyl-amino, C.sub.3-8cycloalkyl-amino,
heterocyclyl-amino (optionally substituted on heterocyclyl with one
or two C.sub.1-8alkyl substituents) and
heterocyclyl-(C.sub.1-8)alkyl-amino (optionally substituted on
heterocyclyl with one or two C.sub.1-8alkyl substituents); [0040]
R.sub.4 is hydrogen or is one, two or three substituents each
selected from the group consisting of C.sub.3-8cycloalkyl, aryl,
heteroaryl and heterocyclyl each optionally substituted with one,
two or three substituents each selected from the group consisting
of C.sub.1-8alkyl, C.sub.1-8alkoxy,
C.sub.1-8alkoxy-(C.sub.1-8)alkyl, cyano, halogen, hydroxy,
halo-(C.sub.1-8)alkyl, halo-(C.sub.1-8)alkoxy,
hydroxy-(C.sub.1-8)alkyl, hydroxy-(C.sub.1-8)alkoxy,
C.sub.1-8alkoxy-carbonyl, amino-(C.sub.1-8)alkyl,
C.sub.1-8alkyl-amino-(C.sub.1-8)alkyl, amino, C.sub.1-8alkyl-amino,
C.sub.1-8alkoxy-carbonyl-amino; [0041] R.sub.2 is selected from the
group consisting of aryl, heteroaryl, benzofused heteroaryl,
heterocyclyl and benzofused heterocyclyl each optionally
substituted with one, two or three substituents each selected from
the group consisting of C.sub.1-8alkyl, C.sub.2-8alkenyl,
C.sub.2-8alkynyl, C.sub.1-8alkoxy, cyano, halogen,
halo-(C.sub.1-8)alkyl, halo-(C.sub.1-8)alkoxy, hydroxy,
heteroaryl-oxy, aryl-oxy (optionally substituted on heteroaryl and
aryl with one, two or three substituents each selected from the
group consisting of C.sub.1-8alkyl, C.sub.1-8alkoxy and halogen);
and [0042] R.sub.3 is hydrogen or is selected from the group
consisting of R.sub.4--(C.sub.1-8)alkyl,
R.sub.4--(C.sub.2-8)alkenyl, R.sub.4--(C.sub.2-8)alkynyl, halogen,
hydroxy, C.sub.3-cycloalkyl, heteroaryl, aryl and heterocyclyl,
optionally substituted on heteroaryl and aryl with one, two or
three substituents each selected from the group consisting of
C.sub.1-8alkyl, C.sub.1-8alkoxy and halogen.
[0043] An example of the present invention is a compound of Formula
(I) and a form thereof wherein R.sub.1 is --N(Ra,Rb).
[0044] An example of the present invention is a compound of Formula
(I) and a form thereof wherein R.sub.1 is --O(Ra).
[0045] An example of the present invention is a compound of Formula
(I) and a form thereof wherein R.sub.1 is
--N(Ra)-(CH.sub.2).sub.p--Ar.sup.1.
[0046] An example of the present invention is a compound of Formula
(I) and a form thereof wherein Ra is hydrogen.
[0047] An example of the present invention is a compound of Formula
(I) and a form thereof wherein Ra is C.sub.1-8alkyl.
[0048] An example of the present invention is a compound of Formula
(I) and a form thereof wherein Rb is selected from the group
consisting of hydrogen and hydroxy-(C.sub.1-8)alkyl.
[0049] An example of the present invention is a compound of Formula
(I) and a form thereof wherein, when R.sub.1 is --N(Ra,Rb), then Ra
and Rb may be taken together with the nitrogen of attachment to
form a heterocyclyl ring having at least one said nitrogen atom,
wherein said ring is optionally substituted with one substituent
selected from the group consisting of C.sub.1-8alkyl, halogen,
hydroxy, hydroxy-(C.sub.1-8)alkyl, amino and
C.sub.1-8alkyl-amino.
[0050] An example of the present invention is a compound of Formula
(I) and a form thereof wherein p is 0, 1 or 2.
[0051] An example of the present invention is a compound of Formula
(I) and a form thereof wherein Ar.sup.1 is selected from the group
consisting of aryl, heteroaryl and heterocyclyl each optionally
substituted with one or two substituents each selected from the
group consisting of R.sub.4--(C.sub.1-8)alkyl,
R.sub.4--(C.sub.1-8)alkoxy, cyano, halogen, nitro,
halo-(C.sub.1-8)alkyl, halo-(C.sub.1-8)alkoxy, hydroxy,
hydroxy-(C.sub.1-8)alkyl, hydroxy-(C.sub.1-8)alkoxy,
C.sub.1-8alkoxy-carbonyl, C.sub.1-8alkoxy-carbonyl-amino,
amino-(C.sub.1-8)alkyl, C.sub.1-8alkyl-amino-(C.sub.1-8)alkyl,
amino, C.sub.1-8alkyl-amino and
heterocyclyl-(C.sub.1-8)alkyl-amino.
[0052] An example of the present invention is a compound of Formula
(I) and a form thereof wherein Ar.sup.1 is selected from the group
consisting of aryl, heteroaryl and heterocyclyl each optionally
substituted with one or two substituents each selected from the
group consisting of R.sub.4--(C.sub.1-8)alkyl,
R.sub.4--(C.sub.1-8)alkoxy, C.sub.1-8alkyl-amino and
heterocyclyl-(C.sub.1-8)alkyl-amino.
[0053] An example of the present invention is a compound of Formula
(I) and a form thereof wherein R.sub.4 is hydrogen or is one, two
or three substituents each selected from the group consisting of
aryl, heteroaryl and heterocyclyl each optionally substituted with
one or two substituents each selected from the group consisting of
C.sub.1-8alkyl, C.sub.1-8alkoxy, halogen, hydroxy,
halo-(C.sub.1-8)alkyl, halo-(C.sub.1-8)alkoxy,
hydroxy-(C.sub.1-8)alkyl, hydroxy-(C.sub.1-8)alkoxy,
amino-(C.sub.1-8)alkyl, C.sub.1-8alkyl-amino-(C.sub.1-8)alkyl,
amino, C.sub.1-8alkyl-amino and C.sub.1-8alkoxy-carbonyl-amino.
[0054] An example of the present invention is a compound of Formula
(I) and a form thereof wherein R.sub.4 is hydrogen or is
heterocyclyl optionally substituted with one or two substituents
each selected from the group consisting of C.sub.1-8alkyl,
C.sub.1-8alkoxy, halogen, hydroxy, hydroxy-(C.sub.1-8)alkyl, amino,
C.sub.1-8alkyl-amino and C.sub.1-8alkoxy-carbonyl-amino.
[0055] An example of the present invention is a compound of Formula
(I) and a form thereof wherein [0056] Ar.sup.1 is selected from the
group consisting of aryl, heteroaryl and heterocyclyl each
optionally substituted with one or two substituents each selected
from the group consisting of R.sub.4--(C.sub.1-8)alkyl,
R.sub.4--(C.sub.1-8)alkoxy, cyano, halogen, nitro,
halo-(C.sub.1-8)alkyl, halo-(C.sub.1-8)alkoxy, hydroxy,
hydroxy-(C.sub.1-8)alkyl, hydroxy-(C.sub.1-8)alkoxy,
C.sub.1-8alkoxy-carbonyl, C.sub.1-8alkoxy-carbonyl-amino,
amino-(C.sub.1-8)alkyl, C.sub.1-8alkyl-amino-(C.sub.1-8)alkyl,
amino, C.sub.1-8alkyl-amino and
heterocyclyl-(C.sub.1-8)alkyl-amino; and [0057] R.sub.4 is hydrogen
or is one, two or three substituents each selected from the group
consisting of aryl, heteroaryl and heterocyclyl each optionally
substituted with one or two substituents each selected from the
group consisting of C.sub.1-8alkyl, C.sub.1-8alkoxy, halogen,
hydroxy, halo-(C.sub.1-8)alkyl, halo-(C.sub.1-8)alkoxy,
hydroxy-(C.sub.1-8)alkyl, hydroxy-(C.sub.1-8)alkoxy,
amino-(C.sub.1-8)alkyl, C.sub.1-8alkyl-amino-(C.sub.1-8)alkyl,
amino, C.sub.1-8alkyl-amino and C.sub.1-8alkoxy-carbonyl-amino.
[0058] An example of the present invention is a compound of Formula
(I) and a form thereof wherein [0059] Ar.sup.1 is selected from the
group consisting of aryl, heteroaryl and heterocyclyl each
optionally substituted with one or two substituents each selected
from the group consisting of R.sub.4--(C.sub.1-8)alkyl,
R.sub.4--(C.sub.1-8)alkoxy, C.sub.1-8alkyl-amino and
heterocyclyl-(C.sub.1-8)alkyl-amino; and [0060] R.sub.4 is hydrogen
or is heterocyclyl optionally substituted with one or two
substituents each selected from the group consisting of
C.sub.1-8alkyl, C.sub.1-8alkoxy, halogen, hydroxy,
hydroxy-(C.sub.1-8)alkyl, amino, C.sub.1-8alkyl-amino and
C.sub.1-8alkoxy-carbonyl-amino.
[0061] An example of the present invention is a compound of Formula
(I) and a form thereof wherein R.sub.2 is selected from the group
consisting of aryl, heteroaryl and benzofused heterocyclyl each
optionally substituted with one, two or three substituents each
selected from the group consisting of C.sub.1-8alkyl,
C.sub.1-8alkoxy, halogen, halo-(C.sub.1-8)alkyl,
halo-(C.sub.1-8)alkoxy, heteroaryl-oxy, aryl-oxy (optionally
substituted on heteroaryl and aryl with one, two or three
substituents each selected from the group consisting of
C.sub.1-8alkyl, C.sub.1-8alkoxy and halogen).
[0062] An example of the present invention is a compound of Formula
(I) and a form thereof wherein R.sub.2 is selected from the group
consisting of aryl, heteroaryl and benzofused heterocyclyl each
optionally substituted with one, two or three substituents each
selected from the group consisting of C.sub.1-8alkoxy, halogen and
aryl-oxy.
[0063] An example of the present invention is a compound of Formula
(I) and a form thereof wherein R.sub.3 is hydrogen or is selected
from the group consisting of R.sub.4--(C.sub.1-8)alkyl,
R.sub.4--(C.sub.2-8)alkenyl, R.sub.4--(C.sub.2-8)alkynyl, halogen,
hydroxy, C.sub.3-cycloalkyl, heteroaryl, aryl and heterocyclyl,
optionally substituted on heteroaryl and aryl with one, two or
three substituents each selected from the group consisting of
C.sub.1-8alkyl, C.sub.1-8alkoxy and halogen.
[0064] An example of the present invention is a compound of Formula
(I) and a form thereof wherein R.sub.3 is hydrogen or is selected
from the group consisting of C.sub.1-8alkyl, halogen, hydroxy, aryl
and heterocyclyl, optionally substituted on aryl with one or two
halogen substituents.
[0065] An example of the present invention is a compound of Formula
(I) and a form thereof wherein [0066] R.sub.1 is selected from the
group consisting of --N(Ra,Rb), --O(Ra) and
--N(Ra)-(CH.sub.2).sub.p--Ar.sup.1; [0067] Ra is selected from the
group consisting of hydrogen and C.sub.1-8alkyl; [0068] Rb is
selected from the group consisting of hydrogen and
hydroxy(C.sub.1-8)alkyl; [0069] alternatively, when R.sub.1 is
--N(Ra,Rb), then Ra and Rb may be taken together with the nitrogen
of attachment to form a heterocyclyl ring having at least one said
nitrogen atom, wherein said ring is optionally substituted with one
substituent selected from the group consisting of C.sub.1-8alkyl,
halogen, hydroxy, hydroxy-(C.sub.1-8)alkyl, amino and
C.sub.1-8alkyl-amino; [0070] p is 0, 1 or 2; [0071] Ar.sup.1 is
selected from the group consisting of aryl, heteroaryl and
heterocyclyl each optionally substituted with one or two
substituents each selected from the group consisting of
R.sub.4--(C.sub.1-8)alkyl, R.sub.4--(C.sub.1-8)alkoxy,
C.sub.1-8alkyl-amino and heterocyclyl-(C.sub.1-8)alkyl-amino;
[0072] R.sub.4 is hydrogen or is heterocyclyl optionally
substituted with one or two substituents each selected from the
group consisting of C.sub.1-8alkyl, C.sub.1-8alkoxy, halogen,
hydroxy, hydroxy-(C.sub.1-8)alkyl, amino, C.sub.1-8alkyl-amino and
C.sub.1-8alkoxy-carbonyl-amino; [0073] R.sub.2 is selected from the
group consisting of aryl, heteroaryl and benzofused heterocyclyl
each optionally substituted with one, two or three substituents
each selected from the group consisting of C.sub.1-8alkoxy, halogen
and aryl-oxy; and [0074] R.sub.3 is hydrogen or is selected from
the group consisting of C.sub.1-8alkyl, halogen, hydroxy, aryl and
heterocyclyl, optionally substituted on aryl with one or two
halogen substituents.
[0075] An example of the present invention is a compound of Formula
(I) and a form thereof, wherein R.sub.1, R.sub.2 and R.sub.3 is
selected from:
TABLE-US-00001 Cpd R.sub.1 R.sub.2 R.sub.3 1 OCH.sub.2CH.sub.3
3-Cl-phenyl OH 2 NH-(4-CH.sub.2-piperidin-1-yl)-phenyl 3-Cl-phenyl
H 3 NH-(4-CH.sub.2-morpholin-4-yl)- 3-Cl-phenyl H phenyl 4
NH-(4-CH.sub.2-morpholin-4-yl)- 5-Cl-benzo[1,3]dioxol-4-yl H phenyl
5 NH-(4-CH.sub.2-morpholin-4-yl)- 2,4-Cl.sub.2-5-OCH.sub.3-phenyl H
phenyl 6 NH-[4-(CH.sub.2).sub.2-morpholin-4-yl]-
2,4-Cl.sub.2-5-OCH.sub.3-phenyl H phenyl 7 OCH.sub.2CH.sub.3
5-Cl-benzo[1,3]dioxol-4-yl morpholin- 4-yl 8
NH-(4-CH.sub.2-morpholin-4-yl)- 5-Cl-benzo[1,3]dioxol-4-yl
morpholin- phenyl 4-yl 9 NH-(4-CH.sub.2-morpholin-4-yl)-
5-Cl-benzo[1,3]dioxol-4-yl CH.sub.3 phenyl 10
NH-(1-CH.sub.3)-piperidin-4-yl 5-Cl-benzo[1,3]dioxol-4-yl H 11
NH-[4-(CH.sub.2).sub.2-piperidin-1-yl]- 5-Cl-benzo[1,3]dioxol-4-yl
H phenyl 12 NH-[4-(CH.sub.2).sub.2-(4-CH.sub.3-piperazin-1-yl)]-
5-Cl-benzo[1,3]dioxol-4-yl H phenyl 13
NH-[4-(CH.sub.2).sub.2-morpholin-4-yl]- 5-Cl-benzo[1,3]dioxol-4-yl
H phenyl 14 OCH.sub.2CH.sub.3 pyridin-4-yl OH 15 OCH.sub.2CH.sub.3
pyridin-4-yl 2-Cl-5-F- phenyl 16
NH--(CH.sub.2).sub.2-pyrrolidin-1-yl pyridin-4-yl 2-Cl-5-F- phenyl
17 OCH.sub.2CH.sub.3 5-Cl-benzo[1,3]dioxol-4-yl H 18
NH-[4-CH.sub.2-(4-OCH.sub.3-piperidin- 5-Cl-benzo[1,3]dioxol-4-yl H
1-yl)]-phenyl 19 NH-[4-CH.sub.2-(4-CH.sub.3-piperidin-1-
5-Cl-benzo[1,3]dioxol-4-yl H yl)]-phenyl 20
NH-[4-CH.sub.2-(3-CH.sub.2OH- 5-Cl-benzo[1,3]dioxol-4-yl H
piperidin-1-yl)]-phenyl 21 3-CH.sub.2OH-piperidin-1-yl
5-Cl-benzo[1,3]dioxol-4-yl H 22 NH-[4-CH.sub.2-(4-CH.sub.2OH-
5-Cl-benzo[1,3]dioxol-4-yl H piperidin-1-yl)]-phenyl 23
NH-[4-CH.sub.2-(4-OH-piperidin-1- 5-Cl-benzo[1,3]dioxol-4-yl H
yl)]-phenyl 24 NH-{4-CH.sub.2-[3,5-(CH.sub.3).sub.2-
5-Cl-benzo[1,3]dioxol-4-yl H morpholin-4-yl]}-phenyl 25
NH-{4-CH.sub.2-[2,6-cis-(CH.sub.3).sub.2-
5-Cl-benzo[1,3]dioxol-4-yl H morpholin-4-yl]}-phenyl 26
NH-[4-CH.sub.2-(4-F-piperidin-1- 5-Cl-benzo[1,3]dioxol-4-yl H
yl)]-phenyl 27 NH-[3-N(CH.sub.3).sub.2-4-CH.sub.2-
5-Cl-benzo[1,3]dioxol-4-yl H morpholin-4-yl]-phenyl 28
4-OH-piperidin-1-yl 5-Cl-benzo[1,3]dioxol-4-yl H 29
N(CH.sub.3)-(4-CH.sub.2-morpholin-4-yl)- 5-Cl-benzo[1,3]dioxol-4-yl
H phenyl 30 NH-[6-NH(CH.sub.2).sub.3-morpholin-4-
5-Cl-benzo[1,3]dioxol-4-yl H yl]-pyridin-3-yl 31
NH-[3-OCH.sub.3-4-O(CH.sub.2).sub.2- 5-Cl-benzo[1,3]dioxol-4-yl H
piperidin-1-yl]-phenyl 32 NH-[4-OCH.sub.2-(1-CH.sub.3-piperidin-
5-Cl-benzo[1,3]dioxol-4-yl H 2-yl)]-phenyl 33 NH-pyrazol-3-yl
5-Cl-benzo[1,3]dioxol-4-yl H 34 NH-(5-CH.sub.3)-isoxazol-3-yl
5-Cl-benzo[1,3]dioxol-4-yl H 35 NH-pyrimidin-2-yl
5-Cl-benzo[1,3]dioxol-4-yl H 36 NH-pyridin-3-yl
5-Cl-benzo[1,3]dioxol-4-yl H 37 NH-pyridin-2-yl
5-Cl-benzo[1,3]dioxol-4-yl H 38 NH--C(CH.sub.3).sub.2CH.sub.2OH
5-Cl-benzo[1,3]dioxol-4-yl H 39 NH-(1-CH.sub.3)-pyrazol-3-yl
5-Cl-benzo[1,3]dioxol-4-yl H 40 NH-pyridin-4-yl
5-Cl-benzo[1,3]dioxol-4-yl H 41 4-N(CH.sub.3).sub.2-piperidin-1-yl
5-Cl-benzo[1,3]dioxol-4-yl H 42
NH-{4-CH.sub.2-[4-N(CH.sub.3).sub.2- 5-Cl-benzo[1,3]dioxol-4-yl H
piperidin-1-yl]}-phenyl 43 3-F-piperidin-1-yl
5-Cl-benzo[1,3]dioxol-4-yl H 44 NH-[4-CH.sub.2-(3-F-piperidin-1-
5-Cl-benzo[1,3]dioxol-4-yl H yl)]-phenyl 45
NH-[3-OCH.sub.3-4-CH.sub.2-morpholin- 4-phenoxy-phenyl H
4-yl]-phenyl 46 NH-{4-CH.sub.2-[4- 5-Cl-benzo[1,3]dioxol-4-yl H
NHC(O)OC(CH.sub.3).sub.3-piperidin-1- yl]}-phenyl 47
NH-[4-CH.sub.2-(4-NH.sub.2-piperidin-1- 5-Cl-benzo[1,3]dioxol-4-yl
H yl)]-phenyl 48 NH-[4-(CH.sub.2).sub.2-piperidin-1-yl]-
4-phenoxy-phenyl H phenyl 49 NH-(4-CH.sub.2-piperidin-1-yl)-
4-phenoxy-phenyl H phenyl 50 NH-(4-CH.sub.2-morpholin-4-yl)-
4-phenoxy-phenyl H phenyl 51 NH-[4-CH.sub.2-(4-OCH.sub.3-piperidin-
4-phenoxy-phenyl H 1-yl)]-phenyl 52 OCH.sub.2CH.sub.3
4-phenoxy-phenyl OH 53 OCH.sub.2CH.sub.3 4-phenoxy-phenyl Cl 54
NH-[4-CH.sub.2-(4,4-F.sub.2-piperidin-1- 5-Cl-benzo[1,3]dioxol-4-yl
H yl)]-phenyl
[0076] An example of the present invention is a compound of Formula
(I) and a form thereof, wherein [0077] R.sub.1 is selected from
OCH.sub.2CH.sub.3, NH-(4-CH.sub.2-piperidin-1-yl)-phenyl,
NH-(4-CH.sub.2-morpholin-4-yl)-phenyl,
NH-[4-(CH.sub.2).sub.2-morpholin-4-yl]-phenyl,
NH-(1-CH.sub.3)-piperidin-4-yl,
NH-[4-(CH.sub.2).sub.2-piperidin-1-yl]-phenyl,
NH-[4-(CH.sub.2).sub.2-(4-CH.sub.3-piperazin-1-yl)]-phenyl,
NH--(CH.sub.2).sub.2-pyrrolidin-1-yl,
NH-[4-CH.sub.2-(4-OCH.sub.3-piperidin-1-yl)]-phenyl,
NH-[4-CH.sub.2-(4-CH.sub.3-piperidin-1-yl)]-phenyl,
NH-[4-CH.sub.2-(3-CH.sub.2OH-piperidin-1-yl)]-phenyl,
3-CH.sub.2OH-piperidin-1-yl,
NH-[4-CH.sub.2-(4-CH.sub.2OH-piperidin-1-yl)]-phenyl,
NH-[4-CH.sub.2-(4-OH-piperidin-1-yl)]-phenyl,
NH-{4-CH.sub.2-[3,5-(CH.sub.3).sub.2-morpholin-4-yl]}-phenyl,
NH-{4-CH.sub.2-[2,6-cis-(CH.sub.3).sub.2-morpholin-4-yl]}-phenyl,
NH-[4-CH.sub.2-(4-F-piperidin-1-yl)]-phenyl,
NH-[3-N(CH.sub.3).sub.2-4-CH.sub.2-morpholin-4-yl]-phenyl,
4-OH-piperidin-1-yl,
N(CH.sub.3)-(4-CH.sub.2-morpholin-4-yl)-phenyl,
NH-[6-NH(CH.sub.2).sub.3-morpholin-4-yl]-pyridin-3-yl,
NH-[3-OCH.sub.3-4-O(CH.sub.2).sub.2-piperidin-1-yl]-phenyl,
NH-[4-OCH.sub.2-(1-CH.sub.3-piperidin-2-yl)]-phenyl,
NH-pyrazol-3-yl, NH-(5-CH.sub.3)-isoxazol-3-yl, NH-pyrimidin-2-yl,
NH-pyridin-3-yl, NH-pyridin-2-yl, NH--C(CH.sub.3).sub.2CH.sub.2OH,
NH-(1-CH.sub.3)-pyrazol-3-yl, NH-pyridin-4-yl,
4-N(CH.sub.3).sub.2-piperidin-1-yl,
NH-{4-CH.sub.2-[4-N(CH.sub.3).sub.2-piperidin-1-yl]}-phenyl,
3-F-piperidin-1-yl, NH-[4-CH.sub.2-(3-F-piperidin-1-yl)]-phenyl,
NH-[3-OCH.sub.3-4-CH.sub.2-morpholin-4-yl]-phenyl,
NH-{4-CH.sub.2-[4-NHC(O)OC(CH.sub.3).sub.3-piperidin-1-yl]}-phenyl,
NH-[4-CH.sub.2-(4-NH.sub.2-piperidin-1-yl)]-phenyl or
NH-[4-CH.sub.2-(4,4-F.sub.2-piperidin-1-yl)]-phenyl; [0078] R.sub.2
is selected from 3-Cl-phenyl, 2,4-Cl.sub.2-5-OCH.sub.3-phenyl,
pyridin-4-yl, 4-phenoxy-phenyl or 5-Cl-benzo[1,3]dioxol-4-yl; and
[0079] R.sub.3 is selected from hydrogen, OH, Cl, morpholin-4-yl,
CH.sub.3 or 2-Cl-5-F-phenyl.
[0080] Compounds representative of a compound of Formula (I) or a
form thereof include compounds and forms thereof selected from:
##STR00004## ##STR00005## ##STR00006## ##STR00007## ##STR00008##
##STR00009## ##STR00010## ##STR00011## ##STR00012## ##STR00013##
##STR00014## ##STR00015## ##STR00016## ##STR00017##
##STR00018##
CHEMICAL DEFINITIONS & NOMENCLATURE
[0081] Bond lines drawn into a ring system from a substituent
variable indicate that the substituent may be attached to any of
the substitutable ring atoms.
[0082] As used herein, the following terms are intended to have the
following definitions. The definitions herein may specify that a
chemical term has an indicated formula. The particular formula
provided is not intended to limit the scope of the invention, but
is provided as an illustration of the term. The scope of the per se
definition of the term is intended to include the plurality of
variations expected to be included by one of ordinary skill in the
art.
[0083] The term "C.sub.1-8alkyl" means a saturated aliphatic
branched or straight-chain hydrocarbon radical or linking group
having from 1 up to 8 carbon atoms in a linear or branched
arrangement, wherein the radical is derived by the removal of one
hydrogen atom from a carbon atom and the linking group is derived
by the removal of one hydrogen atom from each of two carbon atoms
in the chain. The term "C.sub.1-8alkyl" also includes a
"C.sub.1-6alkyl" and "C.sub.1-4alkyl" radical or linking group
having from 1 up to 6 carbon atoms and 1 up to 4 carbon atoms
respectively, such as methyl, ethyl, 1-propyl, 2-propyl, 1-butyl,
2-butyl, tert-butyl, 1-pentyl, 2-pentyl, 3-pentyl, 1-hexyl,
2-hexyl, 3-hexyl, 1-heptyl, 2-heptyl, 3-heptyl, 1-octyl, 2-octyl,
3-octyl and the like. Alkyl radicals may be attached to a core
molecule and further substituted on any atom when allowed by
available valences.
[0084] The term "C.sub.1-8alkoxy" means an alkyl radical or linking
group having from 1 up to 8 carbon atoms in a linear or branched
arrangement, wherein the radical or linking group is attached
through an oxygen linking atom, as in the formula:
--O--C.sub.1-8alkyl. The term "C.sub.1-8alkoxy" also includes a
"C.sub.1-6alkoxy" and "C.sub.1-4alkoxy" radical or linking group
having from 1 up to 6 carbon atoms and from 1 up to 4 carbon atoms
respectively, such as methoxy, ethoxy, propoxy, butoxy and the
like. An alkoxy radical may be attached to a core molecule and
further substituted on any atom when allowed by available
valences.
[0085] The term "C.sub.3-12cycloalkyl" means a saturated or
partially unsaturated cyclic hydrocarbon ring system radical. The
term "C.sub.3-12cycloalkyl" also includes a C.sub.3-8cycloalkyl,
C.sub.3-10cycloalkyl, C.sub.5-6cycloalkyl, C.sub.5-8cycloalkyl,
C.sub.5-12cycloalkyl, C.sub.9-13cycloalkyl or
benzofused-C.sub.3-12cycloalkyl ring system radical and the like,
such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,
cycloheptyl, cyclooctyl, 1H-indenyl, indanyl, 9H-fluorenyl,
1,2,3,4-tetrahydro-naphthalenyl, acenaphthenyl, adamantanyl and the
like. C.sub.3-12cycloalkyl radicals may be attached to a core
molecule and further substituted on any atom when allowed by
available valences.
[0086] The term "aryl" means an unsaturated aromatic hydrocarbon
ring system radical. Aryl ring systems include phenyl,
naphthalenyl, azulenyl, anthracenyl and the like. Examples of aryl
in compounds representative of the present invention include phenyl
or naphthalenyl. Aryl radicals may be attached to a core molecule
and further substituted on any atom when allowed by available
valences.
[0087] The term "hetero", when used as a prefix for a ring system,
refers to the replacement of at least one carbon atom member in the
ring system with a heteroatom selected from N, O, S, S(O), or
SO.sub.2. A hetero ring may have 1, 2, 3 or 4 carbon atom members
replaced by a nitrogen atom. Alternatively, a ring may have 1, 2 or
3 nitrogen atom members and 1 oxygen or sulfur atom member.
Alternatively, a ring may have 1 oxygen or sulfur atom member.
Alternatively, up to two adjacent ring members may be heteroatoms,
wherein one heteroatom is nitrogen and the other heteroatom is
selected from N, S or O.
[0088] The term "heterocyclyl" means a saturated or partially
unsaturated "hetero" ring system radical. Heterocyclyl ring systems
include azetidinyl, 2H-pyrrole, 2-pyrrolinyl, 3-pyrrolinyl,
pyrrolidinyl, 1,3-dioxolanyl, 2-imidazolinyl (also referred to as
4,5-dihydro-1H-imidazolyl), imidazolidinyl, 2-pyrazolinyl,
pyrazolidinyl, tetrazolyl, tetrazolidinyl, piperidinyl,
1,4-dioxanyl, morpholinyl, 1,4-dithianyl, thiomorpholinyl,
piperazinyl, azepanyl, hexahydro-1,4-diazepinyl,
hexahydro-1,4-oxazepanyl, tetrahydro-furanyl, tetrahydro-thienyl,
tetrahydro-pyranyl, tetrahydro-pyridazinyl and the like.
Heterocyclyl radicals may be attached to a core molecule and
further substituted on any atom when allowed by available
valences.
[0089] The term "benzofused heterocyclyl" also includes a
heterocyclyl ring system radical and the like, such as indolinyl
(also referred to as 2,3-dihydro-indolyl), benzo[1,3]dioxolyl,
2,3-dihydro-1,4-benzodioxinyl, 2,3-dihydro-benzofuranyl,
1,2-dihydro-phthalazinyl and the like. Benzofused heterocyclyl
radicals may be attached to a core molecule and further substituted
on any atom when allowed by available valences.
[0090] The term "heteroaryl" means an unsaturated aromatic "hetero"
ring system radical. Heteroaryl ring systems include furyl,
thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl,
isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl,
pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl and the like.
Heteroaryl radicals may be attached to a core molecule and further
substituted on any atom when allowed by available valences.
[0091] The term "benzofused heteroaryl" also includes a heteroaryl
ring system radical and the like, such as indolizinyl, indolyl,
azaindolyl, isoindolyl, benzofuranyl, benzothienyl, indazolyl,
azaindazolyl, benzoimidazolyl, benzothiazolyl, benzoxazolyl,
benzoisoxazolyl, benzothiadiazolyl, benzotriazolyl, purinyl,
4H-quinolizinyl, quinolinyl, isoquinolinyl, cinnolinyl,
phthalazinyl, quinazolinyl, quinoxalinyl, 1,8-naphthyridinyl,
pteridinyl and the like. Benzofused-heteroaryl radicals may be
attached to a core molecule and further substituted on any atom
when allowed by available valences.
[0092] The term "C.sub.1-8alkoxy-(C.sub.1-8)alkyl" means a radical
of the formula: --C.sub.1-8alkyl-O--C.sub.1-8alkyl.
[0093] The term "C.sub.1-8alkoxy-carbonyl" means a radical of the
formula: --C(O)--O--C.sub.1-8alkyl.
[0094] The term "C.sub.1-8alkoxy-carbonyl-amino" means a radical of
the formula: --NH--C(O)--O--C.sub.1-8alkyl or
--N[C(O)--O--C.sub.1-8alkyl].sub.2.
[0095] The term "C.sub.1-8alkyl-amino" means a radical of the
formula: --NH--C.sub.1-8alkyl or --N(C.sub.1-8alkyl).sub.2.
[0096] The term "C.sub.1-8alkyl-amino-(C.sub.1-8)alkyl" means a
radical of the formula: --C.sub.1-8alkyl-NH--C.sub.1-8alkyl,
--C.sub.1-8alkyl-N(C.sub.1-8alkyl).sub.2,
--C.sub.1-8alkyl-NH-(C.sub.1-8alkyl-terminal group),
--C.sub.1-8alkyl-N(C.sub.1-8alkyl-terminal group).sub.2 or
--C.sub.1 s-alkyl-N(C.sub.1-8alkyl)-C.sub.1-8alkyl-terminal
group.
[0097] The term "amino" means a radical of the formula:
--NH.sub.2.
[0098] The term "amino-(C.sub.1-8)alkyl" means a radical of the
formula: --C.sub.1-8alkyl-NH.sub.2.
[0099] The term "aryl-oxy" means a radical of the formula:
--O-aryl.
[0100] The term "C.sub.3-8cycloalkyl-amino" means a radical of the
formula: --NH--C.sub.3-8cycloalkyl or
--N(C.sub.3-8cycloalkyl).sub.2.
[0101] The term "halogen" or "halo" means the group chloro, bromo,
fluoro or iodo.
[0102] The term "halo-(C.sub.1-8)alkoxy" means a radical of the
formula: --O--C.sub.1-8alkyl-(halo).sub.n, wherein one or more
halogen atoms may be substituted on C.sub.1-8alkyl when allowed by
available valences (wherein n represents that amount of available
valences based on the number of carbon atoms in the chain), and
includes monofluoromethoxy, difluoromethoxy, trifluoromethoxy,
trifluoroethoxy and the like.
[0103] The term "halo-(C.sub.1-8)alkyl" means a radical of the
formula: --C.sub.1-8alkyl-(halo).sub.n, wherein one or more halogen
atoms may be substituted on C.sub.1-8alkyl when allowed by
available valences (wherein n represents that amount of available
valences based on the number of carbon atoms in the chain), and
includes monofluoromethyl, difluoromethyl, trifluoromethyl,
trifluoroethyl and the like.
[0104] The term "heteroaryl-oxy" means a radical of the formula:
--O-heteroaryl.
[0105] The term "heterocyclyl-amino" means a radical of the
formula: --NH-heterocyclyl or --N(heterocyclyl).sub.2.
[0106] The term "heterocyclyl-(C.sub.1-8)alkyl-amino" means a
radical of the formula: --NH--C.sub.1-8alkyl-heterocyclyl or
--N(C.sub.1-8alkyl-heterocyclyl).sub.2.
[0107] The term "hydroxy-(C.sub.1-8)alkoxy" means a radical of the
formula: --O--C.sub.1-8alkyl-hydroxy, wherein C.sub.1-8alkyl is
substituted on one or more available carbon chain atoms with one or
more hydroxy radicals when allowed by available valences.
[0108] The term "hydroxy-(C.sub.1-8)alkyl" means a radical of the
formula: --C.sub.1-8alkyl-hydroxy, wherein C.sub.1-8alkyl is
substituted on one or more available carbon chain atoms with one or
more hydroxy radicals when allowed by available valences.
[0109] The term "substituted" means the independent replacement of
one or more hydrogen atoms within a radical with that amount of
substitutents allowed by available valences.
[0110] The term "dependently selected" means that the structure
variables are specified in an indicated combination.
[0111] The term "terminal group" means a moiety attached on a
radical substitutent chain that serves as the last group of atoms
on the chain.
[0112] In general, IUPAC nomenclature rules are used herein.
Compound Forms
[0113] The term "form" means, in reference to compounds of the
present invention, such may exist as, without limitation, a salt,
stereoisomer, tautomer, crystalline, polymorph, amorphous, solvate,
hydrate, ester, prodrug or metabolite form. The present invention
encompasses all such compound forms and mixtures thereof. In an
embodiment the term "form" means salts, in particular
pharmaceutically acceptable salts, stereoisomers, and solvates of
the compounds.
[0114] The term "isolated form" means, in reference to compounds of
the present invention, such may exist in an essentially pure state
such as, without limitation, an enantiomer, a racemic mixture, a
geometric isomer (such as a cis or trans stereoisomer), a mixture
of geometric isomers, and the like. The present invention
encompasses all such compound forms and mixtures thereof.
[0115] The compounds of the invention may be present in the form of
pharmaceutically acceptable salts. For use in medicines, the
"pharmaceutically acceptable salts" of the compounds of this
invention refer to non-toxic acidic/anionic or basic/cationic salt
forms.
[0116] Suitable salt forms include acid addition salts which may,
for example, be formed by mixing a solution of the compound
according to the invention with a solution of an acid such as
acetic acid, adipic acid, benzoic acid, carbonic acid, citric acid,
fumaric acid, glycolic acid, hydrochloric acid, maleic acid,
malonic acid, phosphoric acid, saccharinic acid, succinic acid,
sulphuric acid, tartaric acid, trifluoroacetic acid and the
like.
[0117] Furthermore when the compounds of the present invention
carry an acidic moiety, suitable salts thereof may include alkali
metal salts, e.g. sodium or potassium salts; alkaline earth metal
salts, e.g. calcium or magnesium salts; and salts formed with
suitable organic ligands, e.g. quaternary ammonium salts.
[0118] Thus, representative salts include the following: acetate,
adipate, benzenesulfonate, benzoate, bicarbonate, bisulfate,
bitartrate, borate, bromide, calcium, camsylate (or
camphorsulphonate), carbonate, chloride, clavulanate, citrate,
dihydrochloride, edetate, fumarate, gluconate, glutamate,
glyconate, hydrabamine, hydrobromine, hydrochloride, iodide,
isothionate, lactate, malate, maleate, malonate, mandelate,
mesylate, nitrate, oleate, pamoate, palmitate,
phosphate/diphosphate, saccharinate, salicylate, stearate, sulfate,
succinate, tartrate, tosylate, trichloroacetate, trifluoroacetate
and the like.
[0119] Examples of salt forms of compounds representative of the
present invention include the monohydrochloride salt.
[0120] During any of the processes for preparation of the compounds
of the present invention, it may be necessary and/or desirable to
protect sensitive or reactive groups on any of the molecules
concerned. This may be achieved by means of conventional protecting
groups, such as those described in Protective Groups in Organic
Chemistry, ed. J. F. W. McOmie, Plenum Press, 1973; and T. W.
Greene & P. G. M. Wuts, Protective Groups in Organic Synthesis
3.sup.rd Edition, John Wiley & Sons, 1999. The protecting
groups may be removed at a convenient subsequent stage using
methods known in the art. The scope of the present invention
encompasses all such protected compound forms and mixtures
thereof.
[0121] The invention includes compounds of various isomers and
mixtures thereof. The term "isomer" refers to compounds that have
the same composition and molecular weight but differ in physical
and/or chemical properties. Such substances have the same number
and kind of atoms but differ in structure. The structural
difference may be in constitution (geometric isomers) or in an
ability to rotate the plane of polarized light (optical
isomers).
[0122] The term "optical isomer" means isomers of identical
constitution that differ only in the spatial arrangement of their
groups. Optical isomers rotate the plane of polarized light in
different directions. The term "optical activity" means the degree
to which an optical isomer rotates the plane of polarized
light.
[0123] The term "racemate" or "racemic mixture" means an equimolar
mixture of two enantiomeric species, wherein each isolated specie
rotates the plane of polarized light in the opposite direction such
that the mixture is devoid of optical activity.
[0124] The term "enantiomer" means an isomer having a
nonsuperimposable mirror image. The term "diastereomer" means
stereoisomers that are not enantiomers.
[0125] The term "chiral" means a molecule which, in a given
configuration, cannot be superimposed on its mirror image. This is
in contrast to achiral molecules which can be superimposed on their
mirror images.
[0126] The two distinct mirror image versions of the chiral
molecule are also known as levo (left-handed), abbreviated L, or
dextro (right handed), abbreviated D, depending on which way they
rotate polarized light. The symbols "R" and "S" represent the
configuration of groups around a stereogenic carbon atom(s).
[0127] An example of an enantiomerically enriched form isolated
from a racemic mixture includes a dextrorotatory enantiomer,
wherein the mixture is substantially free of the levorotatory
isomer. In this context, substantially free means the levorotatory
isomer may, in a range, comprise less than 25% of the mixture, less
than 10%, less than 5%, less than 2% or less than 1% of the mixture
according to the formula:
% levorotatory = ( mass levorotatory ) ( mass dextrorotatory ) + (
mass levorotatory ) .times. 100 ##EQU00001##
[0128] Similarly, an example of an enantiomerically enriched form
isolated from a racemic mixture includes a levorotatory enantiomer,
wherein the mixture is substantially free of the dextrorotatory
isomer. In this context, substantially free means the
dextrorotatory isomer may, in a range, comprise less than 25% of
the mixture, less than 10%, less than 5%, less than 2% or less than
1% of the mixture according to the formula:
% dextrotatory = ( mass dextrotatory ) ( mass dextrorotatory ) + (
mass levorotatory ) .times. 100 ##EQU00002##
[0129] The term "geometric isomer" means isomers that differ in the
orientation of substituent atoms in relationship to a carbon-carbon
double bond, to a cycloalkyl ring, or to a bridged bicyclic system.
Substituent atoms (other than hydrogen) on each side of a
carbon-carbon double bond may be in an E or Z configuration. In the
"E" configuration, the substituents are on opposite sides in
relationship to the carbon-carbon double bond. In the "Z"
configuration, the substituents are oriented on the same side in
relationship to the carbon-carbon double bond.
[0130] Substituent atoms (other than hydrogen) attached to a ring
system may be in a cis or trans configuration. In the "cis"
configuration, the substituents are on the same side in
relationship to the plane of the ring; in the "trans"
configuration, the substituents are on opposite sides in
relationship to the plane of the ring. Compounds having a mixture
of "cis" and "trans" species are designated "cis/trans".
[0131] The isomeric descriptors ("R," "S," "E," and "Z") indicate
atom configurations and are intended to be used as defined in the
literature.
[0132] The compounds of the invention may be prepared as individual
isomers by either isomer-specific synthesis or resolved from an
isomeric mixture. Conventional resolution techniques include
combining the free base (or free acid) of each isomer of an
isomeric pair using an optically active acid (or base) to form an
optically active salt (followed by fractional crystallization and
regeneration of the free base), forming an ester or amide of each
of the isomers of an isomeric pair by reaction with an appropriate
chiral auxiliary (followed by fractional crystallization or
chromatographic separation and removal of the chiral auxiliary), or
separating an isomeric mixture of either an intermediate or a final
product using various well known chromatographic methods.
[0133] Furthermore, compounds of the present invention may have at
least one crystalline, polymorph or amorphous form. The plurality
of such forms are intended to be included in the scope of the
invention. In addition, some of the compounds may form solvates
with water (i.e., hydrates) or common organic solvents (e.g.,
organic esters such as ethanolate and the like). The plurality of
such solvates are also intended to be encompassed within the scope
of this invention.
Methods of Use
[0134] The compounds of formula (I) are inhibitors of a protein
kinase such as EGFR, HER-2, c-Src, Lyn, c-Abl, Aurora-A or VEGF,
having an IC.sub.50 (50% inhibition concentration) or an EC.sub.50
(50% effective concentration) in a range of about 50 .mu.M or less,
of about 25 .mu.M or less, of about 15 .mu.M or less, of about 10
.mu.M or less, of about 5 .mu.M or less, of about 1 .mu.M or less,
of about 0.5 .mu.M or less, of about 0.25 .mu.M or less or of about
0.1 .mu.M or less.
[0135] The present invention includes a compound of formula (I) and
forms thereof as a protein kinase inhibitor, wherein the protein
kinase is selected from EGFR, HER-2, c-Src, Lyn, c-Abl, Aurora-A or
VEGF.
[0136] The present invention includes a prodrug form of a compound
of formula (I) and forms thereof as a protein kinase inhibitor.
[0137] The present invention includes a metabolite form of a
compound of formula (I) and forms thereof as a protein kinase
inhibitor.
[0138] The present invention includes an isolated form of a
compound of formula (I) and forms thereof as a protein kinase
inhibitor.
[0139] The present invention includes a compound of formula (I) or
a form thereof, wherein the compound is labeled with a ligand for
use as a marker, and wherein the ligand is a radioligand selected
from deuterium, tritium and the like.
[0140] The present invention includes use of a compound of formula
(I) and forms thereof as an inhibitor of a protein kinase such as
EGFR, HER-2, c-Src, Lyn, c-Abl, Aurora-A or VEGF comprising
contacting the protein kinase domain or receptor with the
compound.
[0141] The present invention includes the use of a compound of
formula (I) and forms thereof as a pharmaceutical composition,
medicine or medicament for treating, preventing or ameliorating a
kinase mediated disease, disorder or condition.
[0142] The present invention includes the use of a compound of
formula (I) and forms thereof as a medicament.
[0143] The present invention includes the use of a compound of
formula (I) and forms thereof in the manufacture of a medicament
for treating, preventing or ameliorating a kinase mediated disease,
disorder or condition.
[0144] The present invention includes the use of a prodrug of a
compound of formula (I) and forms thereof as a pharmaceutical
composition, medicine or medicament for treating, preventing or
ameliorating a kinase mediated disease, disorder or condition.
[0145] The present invention includes the use of a prodrug of a
compound of formula (I) and forms thereof as a medicament.
[0146] The present invention is directed to a method for treating,
preventing or ameliorating a chronic or acute protein kinase
mediated disease, disorder or condition in a subject in need
thereof comprising administering to the subject an effective amount
of a compound of formula (I) and forms thereof.
[0147] The method of the present invention further comprises
administering to the subject an effective amount of a prodrug of a
compound of formula (I) and forms thereof.
[0148] The method of the present invention further comprises
treating, preventing or ameliorating a chronic or acute EGFR,
HER-2, c-Src, Lyn, c-Abl, Aurora-A or VEGF mediated disease,
disorder or condition.
[0149] The method of the present invention wherein the disease,
disorder or condition is associated with increased or unregulated
protein kinase activity, expression or signaling and the like in
the subject.
[0150] The method of the present invention further comprises
administering to the subject an effective amount of a compound of
formula (I) as a pharmaceutical composition, medicine or medicament
thereof.
[0151] The method of the present invention wherein the disease,
disorder or condition is an EGFR kinase mediated head or brain
cancer in the subject, and wherein the compound penetrates the
blood brain barrier.
[0152] The method of the present invention further comprises
treating or ameliorating nerve damage and promoting axon
regeneration subsequent to a brain or spinal cord injury in the
subject, wherein the compound is an EGFR inhibitor.
[0153] The method of the present invention further comprises
treating, preventing or ameliorating viral infection by an EGFR
kinase mediated cytomegalovirus in the subject.
[0154] The term "chronic or acute protein kinase mediated disease,
disorder or condition" as used herein, includes, and is not limited
to diseases, disorders or conditions associated with unregulated
kinase activity and conditions that accompany such activity.
[0155] The term "unregulated protein kinase activity, expression or
signaling" refers to 1) increased or unregulated kinase expression
or signaling, 2) increased kinase expression leading to unregulated
cell proliferation, 3) increased kinase signalling leading to
unregulated cell proliferation, or 4) mutations leading to
constitutive kinase activation. The existence of unregulated kinase
activity may be determined by procedures well known in the art.
[0156] The term "unregulated cell proliferation" refers to cell
proliferation of one or more subset of cells in a multicellular
organism resulting in harm (such as discomfort or decreased life
expectancy) to the multicellular organism.
[0157] Tumor cells which result from unregulated cell proliferation
use many mechanisms to enhance their survival and spread and often
have high rates of proliferation because growth control signals
that keep normal cells in check are defective. Many tumor cells
secrete autocrine growth factors that increase proliferation rates
or they induce other cells to secrete growth factors that they
utilize.
[0158] Tumor cells grow and spread by dislodging from a primary
tumor site, using proteases to digest the extracellular matrix,
spreading in response to migration cues, allowing them to migrate
to certain tissues preferentially where overexpressed adhesion
molecules allow attachment and growth at the new site. The totality
of these and other biological processes are responsible for the
lethal effects of a tumor. A kinase inhibitor may affect one or
more aspects of tumor survival mechanisms and thus be
therapeutically useful. Alternatively, a kinase inhibitor may not
affect one particular tumor survival mechanism but may still be
therapeutically useful by affecting tumor survival by an unknown or
as yet unelucidated mechanism of action.
[0159] The foregoing methods contemplate that a compound of formula
(I) or a form thereof is useful for treating, preventing or
ameliorating diseases, disorders or conditions such as, without
limitation, osteoarthritis, rheumatoid arthritis, synovial pannus
invasion in arthritis, multiple sclerosis, myasthenia gravis,
diabetes mellitus, diabetic angiopathy, diabetic retinopathy,
retinal vessel proliferation, inflammatory bowel disease, Crohns
disease, ulcerative colitis, bone diseases, transplant or bone
marrow transplant rejection, lupus, chronic pancreatitis, cachexia,
septic shock, fibroproliferative and differentiative skin diseases
or disorders, central nervous system diseases, neurodegenerative
diseases, disorders or conditions related to nerve damage and axon
degeneration subsequent to a brain or spinal cord injury, acute or
chronic cancer, occular diseases, viral infections, heart disease,
lung or pulmonary diseases or kidney or renal diseases.
[0160] Certain diseases, disorders or conditions further include,
without limitation, acute or chronic cancer selected from bladder
cancer, brain, head or neck cancer, breast cancer, colorectal
cancer, endometrial cancer, epidermoid cancer, esophageal cancer,
gastric cancer, glioma cancer, lung cancer, ovarian cancer,
pancreatic cancer, prostate cancer, renal cell cancer, Kaposi's
sarcoma, leukemia, lymphoma or papillocarcinoma; and,
cancer-associated pathologies selected from abnormal cell
proliferation, unregulated cell proliferation, tumor growth, tumor
angiopathy, tumor angiogenesis, tumor vascularization or metastatic
cancer cell invasion and migration.
[0161] Certain diseases, disorders or conditions further include,
without limitation, fibroproliferative and differentiative skin
diseases or disorders selected from papilloma formation, psoriasis,
dermatitis, eczema, seborrhea or chemotherapy-induced alopecia;
central nervous system diseases selected from Alzheimer's disease,
Parkinson's disease or depression; occular diseases selected from
macular degeneration, diseases of the cornea or glaucoma; viral
infections selected from mycotic infection, autoimmune disease or
cytomegalovirus; heart disease selected from atherosclerosis,
neointima formation or transplantation-induced vasculopathies such
as arterial restenosis; lung or pulmonary diseases selected from
allergic-asthma, lung fibrosis, pulmonary fibrosis or chronic
obstructive pulmonary disorder; and, kidney or renal diseases
selected from acute, subacute or chronic forms of
glomerulonephritis or membranoproliferative glomerulonephritis,
glomerulosclerosis, congenital multicystic renal dysplasia or
kidney fibrosis.
[0162] Certain HER1 kinase mediated cancer includes, without
limitation, bladder cancer, brain, head or neck cancer, breast
cancer, cervical cancer, colorectal cancer, gastric cancer, glioma
cancer, endometrial cancer, esophageal cancer, lung cancer, ovarian
cancer, pancreatic cancer or renal cell cancer.
[0163] Certain HER2 kinase mediated cancer includes, without
limitation, bladder cancer, brain, head or neck cancer, breast
cancer, colorectal cancer, gastric cancer, endometrial cancer,
esophageal cancer, lung cancer, ovarian cancer, prostate cancer or
renal cell cancer.
[0164] The term "administering," with respect to the methods of the
present invention, refers to a means for treating, ameliorating or
preventing a disease, disorder or syndrome as described herein with
a compound of formula (I) or a form thereof, which would obviously
be included within the scope of the invention albeit not
specifically disclosed for certain of said compounds.
[0165] Such methods include therapeutically or prophylactically
administering an effective amount of compound of formula (I) or a
form thereof at different times during the course of a therapy or
concurrently in a combination form. Such methods further include
administering an effective amount of said compound with one or more
agents at different times during the course of a therapy or
concurrently in a combination form.
[0166] The term "prodrug" means a compound of formula (I) or a form
thereof that is converted in vivo into a functional derivative form
that may contribute to therapeutic biological activity, wherein the
converted form may be: 1) a relatively active form; 2) a relatively
inactive form; 3) a relatively less active form; or, 4) any form
which results, directly or indirectly, from such in vivo
conversions.
[0167] Prodrugs are useful when said compound may be either too
toxic to administer systemically, absorbed poorly by the digestive
tract or broken down by the body before it reaches its target.
Conventional procedures for the selection and preparation of
suitable prodrug derivatives are described in, for example, "Design
of Prodrugs", ed. H. Bundgaard, Elsevier, 1985.
[0168] The term "metabolite" means a prodrug form of a compound of
formula (I) or a form thereof converted by in vivo metabolism or a
metabolic process to a relatively less active functional derivative
of said compound.
[0169] The term "subject" as used herein, refers to a patient, such
as an animal, a mammal or a human, who has been the object of
treatment, observation or experiment and is at risk of (or
susceptible to) developing a disease or disorder or having a
disease or disorder related to unregulated kinase activity.
[0170] The term "effective amount" refers to that amount of a
compound of formula (I) or a form, pharmaceutical composition,
medicine or medicament thereof that elicits the biological or
medicinal response (such as inhibiting activation of unregulated
kinase activity) in a tissue system, animal or human, that is being
sought by a researcher, veterinarian, medical doctor, or other
clinician, which includes alleviation of the symptoms of the
disease or disorder being treated. The effective amount of a
certain compound can be assessed by the skilled person in a
art-known manner.
[0171] The effective amount of said compound is from about 0.001
mg/kg/day to about 300 mg/kg/day.
[0172] The term "pharmaceutical composition" refers to a product
containing a compound of formula (I) or a form thereof, such as a
product comprising the specified ingredients in the specified
amounts, as well as any product which results, directly or
indirectly, from such combinations of the specified ingredients in
the specified amounts.
[0173] The term "medicament" or "medicine" refers to a product
containing a compound of formula (I) or a form thereof. The present
invention includes use of such a medicament for treating,
preventing or ameliorating a chronic or acute kinase mediated
disease, disorder or condition.
[0174] The term "pharmaceutically acceptable" refers to molecular
entities and compositions that are of sufficient purity and quality
for use in the formulation of a pharmaceutical composition,
medicine or medicament of the present invention and that, when
appropriately administered to an animal or a human, do not produce
an adverse, allergic or other untoward reaction. Since both human
use (clinical and over-the-counter) and veterinary use are equally
included within the scope of the present invention, a
pharmaceutically acceptable formulation would include a
pharmaceutical composition, medicine or medicament for either human
or veterinary use.
[0175] The term "combination form" refers to the use of a
combination product comprising a compound of formula (I) or a form,
pharmaceutical composition, medicine or medicament thereof and at
least one therapeutic agent for treating, preventing or
ameliorating a chronic or acute protein kinase mediated disease,
disorder or condition.
[0176] Advantageously, the effective amount of a combination
product for treating, preventing or ameliorating a chronic or acute
protein kinase mediated disease, disorder or condition may be a
reduced amount of either or both the compound or therapeutic agent
compared to the effective amount of the compound or therapeutic
agent otherwise recommended for treating, preventing or
ameliorating the disease, disorder or condition. Therefore, it is
contemplated that the compound is administered to the subject
before, during or after the time the agent is administered.
[0177] The term "therapeutic agent" refers to chemotherapeutic
agents used to treat a kinase mediated cancer or antiviral agents
used to treat cytomegalovirus. Chemotherapeutic agents include and
are not limited to anti-angiogenic agents, anti-tumor agents,
cytotoxic agents, inhibitors of cell proliferation, radiation
therapy and the like or a combination thereof.
[0178] The term "treating, preventing or ameliorating" refers,
without limitation, to facilitating the eradication of, inhibiting
the progression of or promoting stasis of a chronic or acute kinase
mediated disease, disorder or condition.
[0179] The term "radiation therapy" refers to a therapy that
comprises exposing the subject in need thereof to radiation. The
present invention includes a method for administering a compound of
formula (I) or a form, pharmaceutical composition, medicine or
medicament thereof in combination with radiation therapy.
Procedures for administering such therapy are known to those
skilled in the art. The appropriate scheme of radiation therapy
will be similar to those already employed in clinical therapies
wherein the radiation therapy is used alone or in combination with
other chemotherapeutic agents.
[0180] The present invention includes a pharmaceutical composition
comprising an admixture of a compound of formula (I) or a form
thereof and one or more pharmaceutically acceptable excipients.
[0181] The present invention includes a process for making a
pharmaceutical composition, medicine or medicament comprising
mixing a compound of formula (I) or a form thereof and an optional
pharmaceutically acceptable carrier. The present invention includes
a pharmaceutical composition, medicine or medicament resulting from
the process of mixing a compound of formula (I) or a form thereof
and an optional pharmaceutically acceptable carrier. Contemplated
processes include both conventional and unconventional
pharmaceutical techniques.
[0182] Said pharmaceutical composition, medicine or medicament may
take a wide variety of forms to effectuate mode of administration,
wherein the mode includes, and is not limited to, intravenous (both
bolus and infusion), oral, nasal, transdermal, topical with or
without occlusion, and via injection intraperitoneally,
subcutaneously, intramuscularly, intratumorally, intracerebrally or
intracranially. The composition, medicine or medicament may be in a
dosage unit such as a tablet, pill, capsule, powder, granule,
sterile parenteral solution or suspension, metered aerosol or
liquid spray, drop, ampoule, auto-injector device or suppository
for such administration modes.
[0183] Pharmaceutical compositions, medicines or medicaments
suitable for oral administration include solid forms such as pills,
tablets, caplets, capsules (each including immediate release, timed
release and sustained release formulations), granules and powders;
and, liquid forms such as solutions, syrups, elixirs, emulsions and
suspensions. Forms useful for parenteral administration include
sterile solutions, emulsions and suspensions. Alternatively, the
pharmaceutical composition, medicine or medicament may be presented
in a form suitable for once-weekly or once-monthly administration;
for example, an insoluble salt of the active compound, such as the
decanoate salt, may be adapted to provide a depot preparation for
intramuscular injection.
[0184] The dosage form (tablet, capsule, powder, injection,
suppository, teaspoonful and the like) containing the
pharmaceutical composition, medicine or medicament contains an
effective amount of the active ingredient necessary to be
therapeutically or prophylactically effective as described above.
The pharmaceutical composition, medicine or medicament may contain
from about 0.001 mg to about 5000 mg (preferably, from about 0.001
to about 500 mg) of a compound of formula (I) or a form thereof and
may be constituted into any form suitable for the mode of
administration selected for a subject in need.
[0185] An example of a contemplated effective amount for a
pharmaceutical composition, medicine or medicament of the present
invention may range from about 0.001 mg to about 300 mg/kg of body
weight per day. In another example, the range is from about 0.003
to about 100 mg/kg of body weight per day. In another example, the
range is from about 0.005 to about 15 mg/kg of body weight per day.
The pharmaceutical composition, medicine or medicament may be
administered according to a dosage regimen of from about 1 to about
5 times per day.
[0186] For oral administration, the pharmaceutical composition,
medicine or medicament is preferably in the form of a tablet
containing, e.g., 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0,
25.0, 50.0, 100, 150, 200, 250 and 500 milligrams of a compound of
formula (I) or a form thereof for the symptomatic adjustment of the
dosage to the patient to be treated. Optimal dosages will vary
depending on factors associated with the particular patient being
treated (e.g., age, weight, diet and time of administration), the
severity of the condition being treated, the particular compound
being used, the mode of administration and the strength of the
preparation. The use of either daily administration or
post-periodic dosing may be employed.
[0187] A representative compound of formula (I) or a form thereof
includes a compound selected from:
TABLE-US-00002 Cpd Name 1
5-(2-chloro-phenyl)-6-cyano-7-hydroxy-4-oxo-4,5-dihydro-3H-1-thia-3,5,8--
triaza- acenaphthylene-2-carboxylic acid ethyl ester, 2
5-(2-chloro-phenyl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-3,5,8-triaza-
acenaphthylene-2-carboxylic acid
(4-piperidin-1-ylmethyl-phenyl)-amide, 3
5-(2-chloro-phenyl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-3,5,8-triaza-
acenaphthylene-2-carboxylic acid
(4-morpholin-4-ylmethyl-phenyl)-amide, 4
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
(4-morpholin-4-ylmethyl- phenyl)-amide, 5
6-cyano-5-(2,4-dichloro-5-methoxy-phenyl)-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
(4-morpholin-4-ylmethyl- phenyl)-amide, 6
6-cyano-5-(2,4-dichloro-5-methoxy-phenyl)-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(2-morpholin-4-yl-ethyl)- phenyl]-amide, 7
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-7-morpholin-4-yl-4-oxo-4,5-
dihydro-3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid
ethyl ester, 8
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-7-morpholin-4-yl-4-oxo-4,5-
dihydro-3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid (4-
morpholin-4-ylmethyl-phenyl)-amide, 9
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-7-methyl-4-oxo-4,5-dihydro-3H-
- 1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid
(4-morpholin-4- ylmethyl-phenyl)-amide, 10
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
(1-methyl-piperidin-4-yl)- amide, 11
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(2-piperidin-1-yl-ethyl)- phenyl]-amide, 12
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
{4-[2-(4-methyl-piperazin-1- yl)-ethyl]-phenyl}-amide, 13
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(2-morpholin-4-yl-ethyl)- phenyl]-amide, 14
6-cyano-7-hydroxy-4-oxo-5-pyridin-4-yl-4,5-dihydro-3H-1-thia-3,5,8-tria-
za- acenaphthylene-2-carboxylic acid ethyl ester, 15
7-(2-chloro-5-fluoro-phenyl)-6-cyano-4-oxo-5-pyridin-4-yl-4,5-dihydro-3-
H- 1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid ethyl
ester, 16
7-(2-chloro-5-fluoro-phenyl)-6-cyano-4-oxo-5-pyridin-4-yl-4,5-dihydro-3-
H- 1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid
(2-pyrrolidin-1-yl- ethyl)-amide, 17
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid ethyl ester, 18
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(4-methoxy-piperidin-1- ylmethyl)-phenyl]-amide, 19
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(4-methyl-piperidin-1- ylmethyl)-phenyl]-amide, 20
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid [4-(3-hydroxymethyl-
piperidin-1-ylmethyl)-phenyl]-amide, 21
5-(5-chloro-benzo[1,3]dioxol-4-yl)-2-(3-hydroxymethyl-piperidine-1-
carbonyl)-4-oxo-4,5-dihydro-3H-1-thia-3,5,8-triaza-acenaphthylene-6-
carbonitrile, 22
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid [4-(4-hydroxymethyl-
piperidin-1-ylmethyl)-phenyl]-amide, 23
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(4-hydroxy-piperidin-1- ylmethyl)-phenyl]-amide, 24
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(3,5-dimethyl-morpholin-4- ylmethyl)-phenyl]-amide, 25
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid [4-(2,6-cis-dimethyl-
morpholin-4-ylmethyl)-phenyl]-amide, 26
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(4-fluoro-piperidin-1- ylmethyl)-phenyl]-amide, 27
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid (3-dimethylamino-4-
morpholin-4-ylmethyl-phenyl)-amide, 28
5-(5-chloro-benzo[1,3]dioxol-4-yl)-2-(4-hydroxy-piperidine-1-carbonyl)--
4-
oxo-4,5-dihydro-3H-1-thia-3,5,8-triaza-acenaphthylene-6-carbonitrile,
29
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
methyl-(4-morpholin-4- ylmethyl-phenyl)-amide, 30
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid [6-(3-morpholin-4-yl-
propylamino)-pyridin-3-yl]-amide, 31
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
[3-methoxy-4-(2-piperidin-1- yl-ethoxy)-phenyl]-amide, 32
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(1-methyl-piperidin-2- ylmethoxy)-phenyl]-amide, 33
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
(1H-pyrazol-3-yl)-amide, 34
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
(5-methyl-isoxazol-3-yl)- amide, 35
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid pyrimidin-2-ylamide,
36
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid pyridin-3-ylamide, 37
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid pyridin-2-ylamide, 38
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
(2-hydroxy-1,1-dimethyl- ethyl)-amide, 39
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
(1-methyl-1H-pyrazol-3-yl)- amide, 40
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid pyridin-4-ylamide, 41
5-(5-chloro-benzo[1,3]dioxol-4-yl)-2-(4-dimethylamino-piperidine-1-
carbonyl)-4-oxo-4,5-dihydro-3H-1-thia-3,5,8-triaza-acenaphthylene-6-
carbonitrile, 42
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(4-dimethylamino-piperidin- 1-ylmethyl)-phenyl]-amide, 43
5-(5-chloro-benzo[1,3]dioxol-4-yl)-2-(3-fluoro-piperidine-1-carbonyl)-4-
-oxo-
4,5-dihydro-3H-1-thia-3,5,8-triaza-acenaphthylene-6-carbonitrile,
44
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(3-fluoro-piperidin-1- ylmethyl)-phenyl]-amide, 45
6-cyano-4-oxo-5-(4-phenoxy-phenyl)-4,5-dihydro-3H-1-thia-3,5,8-triaza-
acenaphthylene-2-carboxylic acid (3-methoxy-4-morpholin-4-ylmethyl-
phenyl)-amide, 46
[1-(4-{[5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-
-1-
thia-3,5,8-triaza-acenaphthylene-2-carbonyl]-amino}-benzyl)-piperidin-4-y-
l]- carbamic acid tert-butyl ester, 47
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(4-amino-piperidin-1- ylmethyl)-phenyl]-amide, 48
6-cyano-4-oxo-5-(4-phenoxy-phenyl)-4,5-dihydro-3H-1-thia-3,5,8-triaza-
acenaphthylene-2-carboxylic acid
[4-(2-piperidin-1-yl-ethyl)-phenyl]-amide, 49
6-cyano-4-oxo-5-(4-phenoxy-phenyl)-4,5-dihydro-3H-1-thia-3,5,8-triaza-
acenaphthylene-2-carboxylic acid
(4-piperidin-1-ylmethyl-phenyl)-amide, 50
6-cyano-4-oxo-5-(4-phenoxy-phenyl)-4,5-dihydro-3H-1-thia-3,5,8-triaza-
acenaphthylene-2-carboxylic acid
(4-morpholin-4-ylmethyl-phenyl)-amide, 51
6-cyano-4-oxo-5-(4-phenoxy-phenyl)-4,5-dihydro-3H-1-thia-3,5,8-triaza-
acenaphthylene-2-carboxylic acid
[4-(4-methoxy-piperidin-1-ylmethyl)- phenyl]-amide, 52
6-cyano-7-hydroxy-4-oxo-5-(4-phenoxy-phenyl)-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid ethyl ester, 53
7-chloro-6-cyano-4-oxo-5-(4-phenoxy-phenyl)-4,5-dihydro-3H-1-thia-3,5,8-
- triaza-acenaphthylene-2-carboxylic acid ethyl ester, or 54
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(4,4-difluoro-piperidin-1- ylmethyl)-phenyl]-amide.
[0188] A representative compound of formula (I) or a form thereof
includes a compound selected from:
TABLE-US-00003 Cpd Name 3
5-(2-chloro-phenyl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-3,5,8-triaza-
acenaphthylene-2-carboxylic acid
(4-morpholin-4-ylmethyl-phenyl)-amide, 4
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
(4-morpholin-4-ylmethyl-phenyl)- amide, 5
6-cyano-5-(2,4-dichloro-5-methoxy-phenyl)-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
(4-morpholin-4-ylmethyl- phenyl)-amide, 6
6-cyano-5-(2,4-dichloro-5-methoxy-phenyl)-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(2-morpholin-4-yl-ethyl)- phenyl]-amide, 7
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-7-morpholin-4-yl-4-oxo-4,5-
dihydro-3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid
ethyl ester, 8
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-7-morpholin-4-yl-4-oxo-4,5-
dihydro-3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid (4-
morpholin-4-ylmethyl-phenyl)-amide, 9
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-7-methyl-4-oxo-4,5-dihydro-3H-
- 1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid
(4-morpholin-4- ylmethyl-phenyl)-amide, 10
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
(1-methyl-piperidin-4-yl)- amide, 11
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(2-piperidin-1-yl-ethyl)- phenyl]-amide, 12
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
{4-[2-(4-methyl-piperazin-1- yl)-ethyl]-phenyl}-amide, 18
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(4-methoxy-piperidin-1- ylmethyl)-phenyl]-amide, 19
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(4-methyl-piperidin-1- ylmethyl)-phenyl]-amide, 20
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid [4-(3-hydroxymethyl-
piperidin-1-ylmethyl)-phenyl]-amide, 21
5-(5-chloro-benzo[1,3]dioxol-4-yl)-2-(3-hydroxymethyl-piperidine-1-
carbonyl)-4-oxo-4,5-dihydro-3H-1-thia-3,5,8-triaza-acenaphthylene-6-
carbonitrile, 22
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid [4-(4-hydroxymethyl-
piperidin-1-ylmethyl)-phenyl]-amide, 23
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(4-hydroxy-piperidin-1- ylmethyl)-phenyl]-amide, 27
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid (3-dimethylamino-4-
morpholin-4-ylmethyl-phenyl)-amide, 28
5-(5-chloro-benzo[1,3]dioxol-4-yl)-2-(4-hydroxy-piperidine-1-carbonyl)--
4-
oxo-4,5-dihydro-3H-1-thia-3,5,8-triaza-acenaphthylene-6-carbonitrile,
30
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid [6-(3-morpholin-4-yl-
propylamino)-pyridin-3-yl]-amide, 31
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
[3-methoxy-4-(2-piperidin-1- yl-ethoxy)-phenyl]-amide, 32
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(1-methyl-piperidin-2- ylmethoxy)-phenyl]-amide, 33
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
(1H-pyrazol-3-yl)-amide, 34
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
(5-methyl-isoxazol-3-yl)- amide, 35
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid pyrimidin-2-ylamide,
36
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid pyridin-3-ylamide, 40
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid pyridin-4-ylamide, 42
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(4-dimethylamino-piperidin- 1-ylmethyl)-phenyl]-amide, 45
6-cyano-4-oxo-5-(4-phenoxy-phenyl)-4,5-dihydro-3H-1-thia-3,5,8-triaza-
acenaphthylene-2-carboxylic acid (3-methoxy-4-morpholin-4-ylmethyl-
phenyl)-amide, 46
[1-(4-{[5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-
-1-
thia-3,5,8-triaza-acenaphthylene-2-carbonyl]-amino}-benzyl)-piperidin-4-y-
l]- carbamic acid tert-butyl ester, 47
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(4-amino-piperidin-1- ylmethyl)-phenyl]-amide, 48
6-cyano-4-oxo-5-(4-phenoxy-phenyl)-4,5-dihydro-3H-1-thia-3,5,8-triaza-
acenaphthylene-2-carboxylic acid
[4-(2-piperidin-1-yl-ethyl)-phenyl]-amide, 49
6-cyano-4-oxo-5-(4-phenoxy-phenyl)-4,5-dihydro-3H-1-thia-3,5,8-triaza-
acenaphthylene-2-carboxylic acid
(4-piperidin-1-ylmethyl-phenyl)-amide, 50
6-cyano-4-oxo-5-(4-phenoxy-phenyl)-4,5-dihydro-3H-1-thia-3,5,8-triaza-
acenaphthylene-2-carboxylic acid
(4-morpholin-4-ylmethyl-phenyl)-amide, 51
6-cyano-4-oxo-5-(4-phenoxy-phenyl)-4,5-dihydro-3H-1-thia-3,5,8-triaza-
acenaphthylene-2-carboxylic acid
[4-(4-methoxy-piperidin-1-ylmethyl)- phenyl]-amide, or 52
6-cyano-7-hydroxy-4-oxo-5-(4-phenoxy-phenyl)-4,5-dihydro-3H-1-thia-
3,5,8-triaza-acenaphthylene-2-carboxylic acid ethyl ester.
Synthetic Methods
[0189] Representative compounds of the present invention can be
synthesized in accordance with the general synthetic schemes
described below and are illustrated more particularly in the
specific synthetic examples that follow. The general schemes and
specific examples are offered by way of illustration; the invention
should not be construed as being limited by the chemical reactions
and conditions expressed. The methods for preparing the various
starting materials used in the schemes and examples are well within
the skill of persons versed in the art. No attempt has been made to
optimize the yields obtained in any of the example reactions. One
skilled in the art would know how to increase such yields through
routine variations in reaction times, temperatures, solvents and/or
reagents.
[0190] The terms used in describing the invention are commonly used
and known to those skilled in the art. When used herein, the
following abbreviations or formulas have the indicated
meanings:
TABLE-US-00004 Abbreviation Meaning CH.sub.3CN acetonitrile CDI
carbodiimide Cpd compound DIPEA diisopropylethylamine DMF
N,N-dimethyl formamide DMSO dimethyl sulfoxide DME ethylene glycol
dimethyl ether DCM dichloromethane EtOAc ethyl acetate HATU
O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate KOtBu potassium t-butoxide min(s)/hr(s)
minute(s)/hour(s) PyBOP
(benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate
RT/rt/r.t. room temperature TEA or Et.sub.3N triethylamine TFA
trifluoroacetic acid THF tetrahydrofuran
##STR00019##
[0191] 2-cyano-3,3-bis-methylsulfanyl-acrylic acid methyl ester
Compound A1 is reacted with a solution of 2-cyano-thioacetamide
Compound A2 (in a polar solvent such as dimethyl sulfoxide or
ethanol and the like) in the presence of a base (such as potassium
carbonate, sodium bicarbonate, piperidine and the like) to give the
product Compound A3.
[0192] A variation of this method has been described by Elgemeie,
G. H.; Elghandour, A.hmed H.; Elzanate, A. M.; Ahmed, S. A.;
Journal of the Chemical Society, Perkin Transactions 1: Organic and
Bio-Organic Chemistry (1997), (21), 3285-3290.
##STR00020##
[0193] A solution of Compound A3 (in a solvent such as dimethyl
sulfoxide and the like) in the presence of a weak organic base
(such as triethylamine, diisopropylethylamine, N-methylmorpholine
and the like) is reacted with a Compound A4 (wherein Ra is as
previously defined herein), then treated with a solution of
potassium tert-butoxide (in a solvent such as tetrahydrofuran or
tert-butanol and the like) to give a Compound A5.
##STR00021##
[0194] A solution of Compound A5 and an amine Compound A6 (combined
in a polar aprotic solvent such as DMSO and the like) is reacted in
the presence of a non-nucleophilic base such as potassium
tert-butoxide or sodium hydride (in a solvent such as THF,
tert-butanol, DMF and the like) to provide a Compound A7.
##STR00022##
[0195] A solution of Compound A7 (in a solvent such as DMSO, THF
and the like or a mixture thereof) in the presence of a
non-nucleophilic base (such as potassium tert-butoxide and the
like) is reacted with a carbonyl source (such as CDI, phosgene,
triphosgene and the like) to provide a Compound A8, representative
of a compound of Formula (I), wherein R.sub.1 is --O(Ra).
##STR00023##
[0196] Compound A8 is treated with a halogenating reagent (such as
phosphorus oxychloride) to provide a Compound A9 (wherein X is a
halogen atom), representative of a compound of Formula (I), wherein
R.sub.1 is --O(Ra).
##STR00024##
[0197] A solution of Compound A9 (in a solvent such as EtOAc,
ethanol and the like) is reacted under catalytic conditions (with a
catalyst such as palladium on carbon or the like) with a Compound
A10 (wherein Y and R.sub.3 are both hydrogen) in the optional
presence of a base (such as Et.sub.3N, pyridine and the like) to
provide a Compound A11, representative of a compound of Formula
(I), wherein R.sub.1 is --O(Ra).
[0198] Alternatively, Compound A9 may be reacted under transition
metal catalyzed conditions (with a catalyst such as palladium
tetrakis(triphenylphosphine), palladium acetate,
PdCl.sub.2(PPh.sub.3).sub.2 and the like) in the presence of a base
(such as K.sub.2CO.sub.3, NaOtBu, NaOH, pyridine and the like) with
Compound A10 (wherein the Y group for this alternative reaction
sequence represents a group such as a stannane, hydrogen, boronic
acid, boronic ester and the like and R.sub.3 is as previously
defined) to provide the Compound A11, representative of a compound
of Formula (I), wherein R.sub.1 is --O(Ra).
[0199] These reactions employ solvents such as toluene, DME,
1,4-dioxane, water and the like and may require additives such as
phosphine ligands and copper salts.
##STR00025##
[0200] A solution of Compound B1 (in a solvent such as methanol,
ethanol and the like, representative of a compound of Formula (I),
wherein R.sub.1 is --O(Ra) and Ra is alkyl) is reacted with a basic
solution (such as 1N NaOH and the like in a solvent such as
methanol or ethanol and the like) to give Compound B2,
representative of a compound of Formula (I), wherein R.sub.1 is
--O(Ra) and Ra is hydrogen.
##STR00026##
[0201] Compound B2 is reacted with a Compound B3 (using a coupling
reagent such as HATU, PyBOP and the like; wherein Rx represents
NH(Ra,Rb), NH(Ra,Rb) wherein Ra and Rb are taken together with the
nitrogen of attachment to form a heterocyclyl ring and
NH(Ra)-(CH.sub.2).sub.p--Ar.sup.1) to give a Compound B4,
representative of a compound of Formula (I), wherein R.sub.1 is
selected from --N(Ra,Rb), --N(Ra,Rb) wherein Ra and Rb are taken
together with the nitrogen of attachment to form a heterocyclyl
ring and --N(Ra)-(CH.sub.2).sub.p--Ar.sup.1.
[0202] Alternatively, Compound B2 is treated with a reagent (such
as thionyl chloride, oxalyl chloride and the like), then reacted
with a solution of Compound B3 (in a solvent such as THF, DCM and
the like) in the presence of a base (such as DIPEA and the like) to
give a Compound B4, representative of a compound of Formula (I),
wherein R.sub.1 is selected from --N(Ra,Rb), --N(Ra,Rb) wherein Ra
and Rb are taken together with the nitrogen of attachment to form a
heterocyclyl ring and --N(Ra)-(CH.sub.2).sub.p--Ar.sup.1.
Example 1
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-3,5-
,8-triaza-acenaphthylene-2-carboxylic acid
{4-[2-(4-methyl-piperazin-1-yl)-ethyl]-phenyl}-amide (Cpd 12)
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-3,5-
,8-triaza-acenaphthylene-2-carboxylic acid ethyl ester (Cpd 17)
##STR00027##
[0204] 2-cyano-3,3-bis-methylsulfanyl-acrylic acid methyl ester
Compound 1a (25.0 g, 123 mmol) was combined with 97%
2-cyano-thioacetamide (12.7 g, 123 mmol) and K.sub.2CO.sub.3 (51.0
g, 369 mmol) in DMSO (300 mL) at room temperature. The reaction was
stirred for 18 hrs. The cooled reaction mixture (0.degree. C.) was
slowly acidified with 1N HCl. The resultant yellow precipitate was
collected by filtration and sequentially washed with 1N HCl, water,
MeOH and hexane to give
6-hydroxy-4-methylsulfanyl-2-thioxo-1,2-dihydro-pyridine-3,5-dicarbonitri-
le Compound 1b (24.7 g, 90% yield). .sup.1H NMR (DMSOd.sub.6)
.delta. 2.66 (s, 3H). .sup.13C NMR (DMSOd.sub.6) .delta. 118.1
(CN), 117.3 (CN), 17.4 (SMe). MS 222 (M.sup.-).
##STR00028##
[0205] Bromo-acetic acid ethyl ester (also referred to as ethyl
bromoacetate) (4.91 mL, 44.4 mmol) was added dropwise to a mixture
of Compound 1b (9.90 g, 44.4 mmol) and triethylamine (6.19 mL, 44.4
mmol) in DMSO (100 mL). After 30 mins, the reaction was cooled to
10.degree. C. and 1M KOtBu in THF (60 mL) was added dropwise. After
45 mins, the reaction was diluted with 1N HCl and filtered. The
collected yellow solid was washed with 1N HCl, water, MeOH and
hexane to give
3-amino-5-cyano-6-hydroxy-4-methylsulfanyl-thieno[2,3-b]pyridine-2-carbox-
ylic acid ethyl ester Compound 1c (11.1 g). .sup.1H NMR
(DMSOd.sub.6) .delta. 7.28 (br s, 2H), 4.23 (q, 2H), 2.73 (s, 3H),
1.28 (t, 3H). MS 308 (M.sup.-).
##STR00029##
[0206] 1M KOtBu (92.1 mL) was added dropwise to
5-chloro-benzo[1,3]dioxol-4-ylamine hydrochloride salt Compound 1d
(6.36 g, 30.7 mmol) in DMSO (100 mL). After 30 mins, a solution of
Compound 1c (8.80 g, 28.5 mmol) in DMSO (90 mL) was added to the
reaction mixture. The reaction was stirred for 18 hrs and 1N HCl
was added to provide a precipitate. The solid was collected and
washed with 0.5 N NaH.sub.2PO.sub.4, MeOH and hexane to give
3-amino-4-(5-chloro-benzo[1,3]dioxol-4-ylamino)-5-cyano-6-hydroxy-thieno[-
2,3-b]pyridine-2-carboxylic acid ethyl ester Compound 1e (8.81 g).
.sup.1H NMR (DMSOd.sub.6) .delta. 7.25 (br s, 2H), 7.03 (d, 1H),
6.95 (d, 1H), 6.14 (s, 2H), 4.23 (q, 2H), 1.28 (t, 3H). MS 433, 435
(MH.sup.+).
##STR00030##
[0207] 1M KOtBu (41.7 mL) was added dropwise to Compound 1e (6.02
g, 13.9 mmol) in DMSO (150 mL) and stirred for 1 hr. CDI (10.0 g,
61.6 mmol) was then added in one portion. Reaction progress was
monitored by HPLC. After 18 hrs, the reaction was diluted with
EtOAc and 1N HCl and filtered. The filtrate cake was rinsed and the
mother liquor was extracted with EtOAc. The organic layer was dried
(MgSO.sub.4), then filtered and evaporated to give
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-7-hydroxy-4-oxo-4,5-dihyd-
ro-3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid ethyl
ester Compound 1f (5.94 g). .sup.1H NMR (DMSOd.sub.6) .delta. 7.18
(d, 1H), 7.13 (d, 1H), 6.20 (s, 2H, rotomers), 4.31 (q, 2H), 1.30
(t, 3H). LC/MS 459, 461 (MH.sup.+).
##STR00031##
[0208] Compound 1f (2.6 g, 5.68 mmol) was combined with phosphorus
oxychloride (40 mL) in sealed microwave reactor vials and heated to
200.degree. C. for 1 hr. The reaction mixture was slowly added to
ice water and then extracted with EtOAc. The organic layer was
washed with sat'd NaHCO.sub.3, then dried over MgSO.sub.4 and
evaporated. The resultant solid was recrystallized from
EtOAc/hexane to give
7-chloro-5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H--
1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid ethyl ester
Compound 1g (2.19 g). .sup.1H NMR (DMSOd.sub.6) .delta. 7.21 (d,
1H), 7.18 (d, 1H), 6.23 (s, 2H, rotomers), 4.39 (q, 2H), 1.32 (t,
3H). LC/MS 477, 479, 481 (MH.sup.+).
##STR00032##
[0209] A mixture of Compound 1g (1.70 g, 3.57 mmol) in pyridine (5
mL) and EtOAc (50 mL) was combined with 10% Pd/C (6.8 g) in a Parr
shaker and reacted under 40 PSI hydrogen for 2 hrs. The product was
filtered through Celite 545 and evaporated to give Compound 17
(1.56 g) as a yellow solid. .sup.1H NMR (DMSOd.sub.6) .delta. 8.82
(s, 1H) 7.22 (d, 1H), 7.19 (d, 1H), 6.22 (s, 2H, rotomers), 4.39
(q, 2H), 1.33 (t, 3H). LC/MS 443, 445 (MH.sup.+).
##STR00033##
[0210] Compound 17 (1.24 g, 2.81 mmol) was refluxed for 2 hrs in 1N
NaOH (10 mL) and EtOH (50 mL). The reaction was cooled to RT and
partitioned between 1N HCl and EtOAc. The aqueous layer was
extracted with EtOAc, dried (MgSO.sub.4) and evaporated, then
precipitated from EtOAc/MeOH to provide
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro-3H-1-
-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid Compound 1h
(732 mg). LC/MS 415, 417 (MH.sup.+).
##STR00034##
[0211] Compound 1h (152 mg, 0.367 mmol) was refluxed in thionyl
chloride (5 mL) for 1 hr. The excess thionyl chloride was
evaporated in vacuo and then further evaporated from DCM to provide
a solid. 4-[2-(4-methyl-piperazin-1-yl)-ethyl]-phenylamine Compound
1i (80 mg, 0.367 mmol) was added to a solution of the solid in THF
(5 mL) and DIPEA (0.26 mL, 1.47 mmol). After 4 hrs, the reaction
mixture was diluted with sat'd NaHCO.sub.3 and extracted with
EtOAc. The product was dried (MgSO.sub.4) and evaporated, then
purified by reverse phase chromatography (C18,
CH.sub.3CN/water/0.05% TFA gradient) to give Compound 12 (61 mg) as
a white solid. .sup.1H NMR (DMSOd.sub.6) .delta. 8.79 (s, 1H) 7.64
(d, 2H), 7.28 (d, 2H), 7.22 (d, 1H), 7.18 (d, 1H), 6.22 (s, 2H,
rotomers), 4.00-2.79 (m, 12H), 2.76 (s, 3H). LC/MS 616, 618
(MH.sup.+).
[0212] Using the procedure of Example 1, other representative
compounds of the present invention and forms thereof may be
prepared including, but not limited to:
TABLE-US-00005 MS** Cpd Name (MH.sup.+) 1
5-(2-chloro-phenyl)-6-cyano-7-hydroxy-4-oxo-4,5-dihydro-3H-1- 415,
417 thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid ethyl ester,
2 5-(2-chloro-phenyl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-3,5,8-
543, 545 triaza-acenaphthylene-2-carboxylic acid
(4-piperidin-1-ylmethyl-phenyl)- amide, 3
5-(2-chloro-phenyl)-6-cyano-4-oxo-4,5-dihydro-3H-1-thia-3,5,8- 545,
547 triaza-acenaphthylene-2-carboxylic acid (4-morpholin-4-
ylmethyl-phenyl)-amide, 4
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 589,
591 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid (4-
morpholin-4-ylmethyl-phenyl)-amide, 5
6-cyano-5-(2,4-dichloro-5-methoxy-phenyl)-4-oxo-4,5-dihydro- 609,
611 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid (4-
morpholin-4-ylmethyl-phenyl)-amide, 6
6-cyano-5-(2,4-dichloro-5-methoxy-phenyl)-4-oxo-4,5-dihydro- 623,
625 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid [4-(2-
morpholin-4-yl-ethyl)-phenyl]-amide, 10
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 511,
513 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid (1-
methyl-piperidin-4-yl)-amide, 11
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 601,
603 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid [4-(2-
piperidin-1-yl-ethyl)-phenyl]-amide, 13
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 603,
605 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid [4-(2-
morpholin-4-yl-ethyl)-phenyl]-amide, 14
6-cyano-7-hydroxy-4-oxo-5-pyridin-4-yl-4,5-dihydro-3H-1-thia- 382,
384 3,5,8-triaza-acenaphthylene-2-carboxylic acid ethyl ester, 18
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 617,
619 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid [4-(4-
methoxy-piperidin-1-ylmethyl)-phenyl]-amide, 19
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 601,
603 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid [4-(4-
methyl-piperidin-1-ylmethyl)-phenyl]-amide, 20
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 617,
619 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid [4-(3-
hydroxymethyl-piperidin-1-ylmethyl)-phenyl]-amide, 21
5-(5-chloro-benzo[1,3]dioxol-4-yl)-2-(3-hydroxymethyl- 512, 514
piperidine-1-carbonyl)-4-oxo-4,5-dihydro-3H-1-thia-3,5,8-triaza-
acenaphthylene-6-carbonitrile, 22
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 617,
619 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid [4-(4-
hydroxymethyl-piperidin-1-ylmethyl)-phenyl]-amide, 23
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 603,
605 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid [4-(4-
hydroxy-piperidin-1-ylmethyl)-phenyl]-amide, 24
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 617,
619 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(3,5- dimethyl-morpholin-4-ylmethyl)-phenyl]-amide, 25
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 617,
619 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(2,6- cis-dimethyl-morpholin-4-ylmethyl)-phenyl]-amide, 26
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 605,
607 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid [4-(4-
fluoro-piperidin-1-ylmethyl)-phenyl]-amide, 27
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 632,
634 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid (3-
dimethylamino-4-morpholin-4-ylmethyl-phenyl)-amide, 28
5-(5-chloro-benzo[1,3]dioxol-4-yl)-2-(4-hydroxy-piperidine-1- 498,
500 carbonyl)-4-oxo-4,5-dihydro-3H-1-thia-3,5,8-triaza-
acenaphthylene-6-carbonitrile, 29
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 603,
605 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid methyl-
(4-morpholin-4-ylmethyl-phenyl)-amide, 30
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 633,
635 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid [6-(3-
morpholin-4-yl-propylamino)-pyridin-3-yl]-amide, 31
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 647,
649 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid [3-
methoxy-4-(2-piperidin-1-yl-ethoxy)-phenyl]-amide, 32
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 617,
619 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid [4-(1-
methyl-piperidin-2-ylmethoxy)-phenyl]-amide, 33
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 480,
482 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid (1H-
pyrazol-3-yl)-amide, 34
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 495,
497 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid (5-
methyl-isoxazol-3-yl)-amide, 35
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 492,
494 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid
pyrimidin-2-ylamide, 36
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 491,
493 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid
pyridin- 3-ylamide, 37
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 491,
493 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid
pyridin- 2-ylamide, 38
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 486,
488 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid (2-
hydroxy-1,1-dimethyl-ethyl)-amide, 39
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 494,
496 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid (1-
methyl-1H-pyrazol-3-yl)-amide, 40
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 491,
493 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid
pyridin- 4-ylamide, 41
5-(5-chloro-benzo[1,3]dioxol-4-yl)-2-(4-dimethylamino- 525, 527
piperidine-1-carbonyl)-4-oxo-4,5-dihydro-3H-1-thia-3,5,8-triaza-
acenaphthylene-6-carbonitrile, 42
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 630,
632 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid [4-(4-
dimethylamino-piperidin-1-ylmethyl)-phenyl]-amide, 43
5-(5-chloro-benzo[1,3]dioxol-4-yl)-2-(3-fluoro-piperidine-1- 500,
502 carbonyl)-4-oxo-4,5-dihydro-3H-1-thia-3,5,8-triaza-
acenaphthylene-6-carbonitrile, 44
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 605,
607 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid [4-(3-
fluoro-piperidin-1-ylmethyl)-phenyl]-amide, 45
6-cyano-4-oxo-5-(4-phenoxy-phenyl)-4,5-dihydro-3H-1-thia- 633, 635
3,5,8-triaza-acenaphthylene-2-carboxylic acid (3-methoxy-4-
morpholin-4-ylmethyl-phenyl)-amide, 46
[1-(4-{[5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5- 702,
704 dihydro-3H-1-thia-3,5,8-triaza-acenaphthylene-2-carbonyl]-
amino}-benzyl)-piperidin-4-yl]-carbamic acid tert-butyl ester, 47
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 602,
604 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid [4-(4-
amino-piperidin-1-ylmethyl)-phenyl]-amide, 48
6-cyano-4-oxo-5-(4-phenoxy-phenyl)-4,5-dihydro-3H-1-thia- 615, 617
3,5,8-triaza-acenaphthylene-2-carboxylic acid [4-(2-piperidin-1-
yl-ethyl)-phenyl]-amide, 49
6-cyano-4-oxo-5-(4-phenoxy-phenyl)-4,5-dihydro-3H-1-thia- 601, 603
3,5,8-triaza-acenaphthylene-2-carboxylic acid (4-piperidin-1-
ylmethyl-phenyl)-amide, 50
6-cyano-4-oxo-5-(4-phenoxy-phenyl)-4,5-dihydro-3H-1-thia- 603, 605
3,5,8-triaza-acenaphthylene-2-carboxylic acid (4-morpholin-4-
ylmethyl-phenyl)-amide, 51
6-cyano-4-oxo-5-(4-phenoxy-phenyl)-4,5-dihydro-3H-1-thia- 631, 633
3,5,8-triaza-acenaphthylene-2-carboxylic acid [4-(4-methoxy-
piperidin-1-ylmethyl)-phenyl]-amide, 52
6-cyano-7-hydroxy-4-oxo-5-(4-phenoxy-phenyl)-4,5-dihydro-3H- 473,
475 1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid ethyl
ester, 53
7-chloro-6-cyano-4-oxo-5-(4-phenoxy-phenyl)-4,5-dihydro-3H- 491,
493 1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid ethyl
ester, or 54
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihydro- 623,
625 3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid
[4-(4,4- difluoro-piperidin-1-ylmethyl)-phenyl]-amide. **The two
numbers for the MH.sup.+ is due to the fact that the molecules
contain chlorine and the chlorine has two isotopes with substantial
abundance, i.e. .sup.35Cl and .sup.37Cl.
Example 2
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-7-methyl-4-oxo-4,5-dihydro-3H-1-
-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid
(4-morpholin-4-ylmethyl-phenyl)-amide (Cpd 9)
##STR00035##
[0214]
7-chloro-5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-4-oxo-4,5-dihyd-
ro-3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid ethyl
ester Compound 1g (115 mg, 0.242 mmol) was combined with 95%
tetramethyltin (0.039 ml, 0.266 mmol), PdCl.sub.2(Ph.sub.3P).sub.2
(17 mg, 0.024 mmol) and DMF (3 mL) in a sealed microwave vessel.
The mixture was heated to 150.degree. C. for 15 mins. The reaction
mixture was diluted with 0.5 M NaH.sub.2PO.sub.4 and extracted with
EtOAc, then dried (MgSO.sub.4) and evaporated. The resulting
residue was purified by flash chromatography (30 to 50%
EtOAc/hexane/1% Et.sub.3N) to give
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-7-methyl-4-oxo-4,5-dihydro-3H--
1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid ethyl ester
Compound 2a (57 mg) as a white solid. LC/MS 457 (MH.sup.+).
##STR00036##
[0215] Using the procedure of Example 1, Compound 2a was used in
place of Compound 17 and carried forward, then
4-morpholin-4-ylmethyl-phenylamine Compound 2b was used in place of
4-[2-(4-methyl-piperazin-1-yl)-ethyl]-phenylamine Compound 1i to
give Compound 9 as an off-white solid. .sup.1H NMR (DMSOd.sub.6)
.delta. 10.38 (s, 1H), 10.0 (br s, 1H) 7.79 (d, 2H), 7.49 (d, 2H),
7.20 (d, 1H), 7.15 (d, 1H) 6.21 (s, 2H, rotomers), 4.32 (s, 2H),
4.09-3.00 (m, 8H), 2.65 (s, 3H). LC/MS 603, 605 (MH.sup.+).
Example 3
7-(2-chloro-5-fluoro-phenyl)-6-cyano-4-oxo-5-pyridin-4-yl-4,5-dihydro-3H-1-
-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid ethyl ester
(Cpd 15)
7-(2-chloro-5-fluoro-phenyl)-6-cyano-4-oxo-5-pyridin-4-yl-4,5-dihydro-3H-1-
-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid
(2-pyrrolidin-1-yl-ethyl)-amide (Cpd 16)
##STR00037##
[0217]
7-chloro-6-cyano-4-oxo-5-pyridin-4-yl-4,5-dihydro-3H-1-thia-3,5,8-t-
riaza-acenaphthylene-2-carboxylic acid ethyl ester Compound 3a (537
mg, 1.35 mmol), prepared using the procedure of Example 1, was
combined with 2-chloro-5-fluorophenyl-boronic acid (468 mg, 2.69
mmol), Pd(Ph.sub.3P).sub.4 (156 mg, 0.135 mmol), sat'd NaHCO.sub.3
(15 mL) and 1,4-dioxane (40 mL). The reaction mixture was refluxed
for 2 hrs then diluted with sat'd NaHCO.sub.3 and extracted with
EtOAc. The organic layer was dried (MgSO.sub.4) and evaporated. The
solid was recrystallized from EtOAc to give
7-(2-chloro-5-fluoro-phenyl)-6-cyano-4-oxo-5-pyridin-4-yl-4,5-dihydro-3H--
1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid ethyl ester
Compound 15 (440 mg) as a pale yellow solid. LC/MS 494, 496
(MH.sup.+).
##STR00038##
[0218] Compound 15 (394 mg, 0.80 mmol) was refluxed for 2 hrs in 1N
NaOH (5 mL), THF (5 mL) and EtOH (5 mL). The reaction mixture was
cooled to RT and diluted with 0.5 M NaH.sub.2PO.sub.4 to provide a
precipitate, which was collected and used without further
purification. LC/MS 466, 468 (MH.sup.+). The crude solid (67 mg,
0.144 mmol) was combined with HATU (66 mg, 0.173 mmol) in THF (3
mL) and stirred for 30 mins.
[0219] DIPEA (0.03 ml, 0.173 mmol) and 2-pyrrolidin-1-yl-ethylamine
Compound 3c (20 mg, 0.173 mmol) were added to the reaction mixture.
After 4 hrs, the reaction was diluted with 1M NaOH and extracted
with EtOAc, then dried (MgSO.sub.4) and evaporated. The crude
product was purified by reverse phase chromatography (C18,
CH.sub.3CN/water/0.05% TFA gradient) to give Compound 16 (3 mg) as
a white solid. LC/MS 562, 564 (MH.sup.+).
Example 4
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-7-morpholin-4-yl-4-oxo-4,5-dihy-
dro-3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid ethyl
ester (Cpd 7)
5-(5-chloro-benzo[1,3]dioxol-4-yl)-6-cyano-7-morpholin-4-yl-4-oxo-4,5-dihy-
dro-3H-1-thia-3,5,8-triaza-acenaphthylene-2-carboxylic acid
(4-morpholin-4-ylmethyl-phenyl)-amide (Cpd 8)
##STR00039##
[0221] Compound 1g (108 mg, 0.227 mmol) was combined with
morpholine (0.040 ml, 0.454 mmol) and EtOH (3 mL) in a sealed
microwave vial. The mixture was heated in a microwave at
120.degree. C. for 15 mins. The reaction mixture was diluted with
0.5 M NaH.sub.2PO.sub.4 and the precipitate collected by
filtration. The crude solid was washed with MeOH and hexane to give
Compound 7 (89 mg). .sup.1H NMR (DMSOd.sub.6) .delta. 11.28 (s,
1H), 7.16 (d, 1H), 7.14 (d, 1H), 6.19 (s, 2H, rotomers), 4.33 (q,
2H), 3.70-3.63 (m, 4H), 3.48-3.43 (m, 4H), 1.30 (t, 3H). LC/MS 528,
530 (MH.sup.+).
##STR00040##
[0222] Using the procedure of Example 1, Compound 7 was used in
place of Compound 17 and carried forward, then
4-morpholin-4-ylmethyl-phenylamine Compound 2b was used in place of
4-[2-(4-methyl-piperazin-1-yl)-ethyl]-phenylamine Compound 1i to
give Compound 8 as an off-white TFA salt. .sup.1H NMR (DMSOd.sub.6)
.delta. 10.23 (s, 1H), 9.92 (br s, 1H) 7.78 (d, 2H), 7.48 (d, 2H),
7.17 (d, 1H), 7.14 (d, 1H) 6.20 (s, 2H, rotomers), 4.30 (br s, 2H),
4.08-3.88 (m, 2H), 3.73-3.02 (m, 14H). LC/MS 674, 676
(MH.sup.+).
BIOLOGICAL EXAMPLES
[0223] The usefulness of the compounds of the present invention for
treating, preventing or ameliorating a chronic or acute kinase
mediated disease, disorder or condition was determined using the
following procedures.
[0224] Examples 6 and 8 are intended as prophetic examples and are
expected to demonstrate that said compounds are useful in treating,
preventing or ameliorating a chronic or acute kinase mediated
disease, disorder or condition as an inhibitor of the indicated
kinase.
Example 1
EGFR Kinase Assay
[0225] The EGFR kinase used was a fusion of
Glutathione-S-Transferase (GST) and a PCR amplified intracellular
portion of EGFR (NM.sub.--005228). The intracellular portion of
EGFR started at nucleotide 2189 (corresponding to amino acid 667)
and ended at the termination codon. The portion was PCR amplified
with primers that added the lambda attB sequences to each end,
recombined into an entry vector, then into a GST destination vector
(as described in Gateway Technologies Manual by Invitrogen
Corporation, Carlsbad, Calif.).
[0226] The destination vector was recombined in the DH10BAC strain
of bacteria to produce a bacmid. The bacmid was transfected into Sf
9 cells and the supernatant containing the baculovirus was
collected. The GSTEGFR protein was purified using large cultures of
Sf 9 cells infected with stock virus. After an appropriate period
of time, the cells were collected and lysed. The GSTEGFR was then
purified from the lysate on Glutathione-Sepharose columns (as
described by Amersham Biosciences, Buckinghamshire, United
Kingdom).
[0227] The EGFR substrate was prepared by biotinylating polyGluTyr
(128 mg) (Sigma, St. Louis, Mo.) in a 1.times.PBS buffer incubated
together with a 12-fold molar excess of Sulfo-NHS-LC-Biotin on ice
for at least 2 hrs. The free biotin was separated from the
biotinylated polyGluTyr on a gel filtration column.
[0228] A mixture of a 10.times. kinase buffer (500 mM Tris at pH
8.0, 100 mM Magnesium Chloride and 1 mM Sodium Vanadate), DTT (1 mM
final from 500 mM stock), ATP (5 .mu.M final from 10 mM stock),
biotinylated polyGluTyr (10 .mu.g/.mu.L stock), .gamma.-.sup.33P
ATP (10 .mu.Ci/.mu.L stock) and water was added to each well (90
.mu.L/well) of a Streptavidin Flashplate (Perkin Elmer, Wellesley,
Mass.).
[0229] Test compound in 100% DMSO (2 .mu.L) was added to the
appropriate wells. Diluted GSTEGFR (1:300 dilution in 50 mM Tris at
pH 8.0 and 0.1% bovine serum albumin) (10 .mu.L) was added to the
wells to initiate the reactions.
[0230] The plates were incubated at 30.degree. C. for 1 hr with
shaking. The reacted contents were removed and the plates were
sequentially washed three times with a 1.times.PBS stop buffer (300
.mu.L without Magnesium and Calcium) and 100 mM EDTA. After the
final wash, the same stop buffer (200 .mu.L) was added to the
wells. The plates were then sealed and read on the TopCount
scintillation counter.
[0231] Test compounds were assayed in triplicate at 16
concentrations at half-log dilutions starting at 200 uM. A maximum
and minimum signal for the assay was determined on each plate. The
percent inhibition of a test compound was calculated according to
the formula
[ ( max signal - test compound ) ( max signal - min signal ) ] (
100 ) = % inhibition ##EQU00003##
[0232] For a series of test concentrations, the IC.sub.50 was
derived by graphing percent inhibition against the log of the
concentrations tested for a given compound. The IC.sub.50 results
are shown in Table 1. For those compounds without an IC.sub.50, the
percent inhibition results are shown at a test concentration of 2
.mu.M.
TABLE-US-00006 TABLE 1 EGFR IC.sub.50 (.mu.M) Cpd IC.sub.50 (avg) 2
1.67 3 2.21 4 36% 5 1% 6 7% 7 -5% 8 -6% 9 14% 10 3.96 11 1.60 12
2.70 13 23% 14 -9% 15 6% 18 38% 19 0.624 20 0.910 21 32% 22 0.567
23 0.598 24 24% 25 24% 26 2.02 27 34% 28 9% 29 2% 30 11% 31 0.513
32 35% 33 1.22 34 30% 35 42% 36 35% 37 10% 38 13% 39 26% 40 30% 41
7% 42 0.850 45 0.093 46 0.152 47 0.514 48 39% 49 0.224 50 0.124 51
48% 52 0.0169 53 2% 54 11%
Example 2
VEGF-R.sub.2 and Aurora-A Screening Assays
[0233] A kinase reaction mixture was prepared containing 50 mM
Tris-HCl at pH 8, 10 mM MgCl.sub.2, 0.1 mM Na.sub.3PO.sub.4, 1 mM
DTT, 10 .mu.M ATP, 0.025 .mu.M biotinylated histone-H1 peptide
substrate and 0.2 .mu.Curies per well .sup.33P-.gamma.-ATP
(2000-3000 Ci/mmol). 70 .mu.L of the kinase reaction mixture was
dispensed into the well of a Streptavidin FlashPlate.
[0234] Test compound stock in 100% DMSO (1 .mu.L) was added to the
wells resulting in a final concentration of 1% DMSO in the reaction
with a 100 .mu.L final reaction volume. Each enzyme was diluted in
50 mM Tris-HCl pH=8.0, 0.1% BSA and 30 .mu.L was added to each well
to initiate the reaction. The reaction was incubated for one hour
at 30.degree. C. At the end of the 1 hr incubation, the reaction
was terminated by aspirating the mixture from the plate and washing
the wells twice with PBS containing 100 mM EDTA. The biotinylated
peptide substrate became immobilized on the Flashplate.TM.u and the
incorporation of .sup.33P-.gamma.-ATP was measured by reading the
plate on a scintillation counter. Inhibition of the enzymatic
activity was measured by observing a reduced amount of
.sup.33P-.gamma.-ATP incorporated into the immobilized peptide.
[0235] The VEGF-R.sub.2 enzyme is a fusion protein containing a
polyhistidine tag at the N terminus followed by amino acids 786 to
1343 of the rat VEGF-R.sub.2 kinase domain (Accession number
U93306). The assay used 150 ng of the N-terminal biotinylated
peptide biotin-KHKKLAEGSAYEEV-amide (VEGF-R.sub.2) per well.
[0236] Aurora-A is a fusion protein containing a polyhistidine tag
at the N terminus followed by the full length protein encoding the
murine Aurora-A (Accession number GB BC014711) expressed and
purified from sf9 insect cells. The assay used 400 ng of the
N-terminal biotinylated peptide biotin-GRTGRRNSI-amide (Aurora-A)
per well.
[0237] The IC.sub.50 was derived according to the procedure
described in Example 2. The IC.sub.50 results for VEGF-R.sub.2 are
shown in Table 2A and the IC.sub.50 results for Aurora-A are shown
in Table 2B. For those compounds without an IC.sub.50, the percent
inhibition results are shown at a test concentration of 1
.mu.M.
TABLE-US-00007 TABLE 2A VEGF-R2 IC.sub.50 (.mu.M) Cpd IC.sub.50
(avg) 2 >100 3 >100 4 >10 5 >100 6 >100 7 >100 8
14.1 9 >10 10 ~100 11 >100 12 >100 13 >10 14 >100 15
>100 18 >10 19 >100 20 24% 21 >100 22 >100 23 28% 24
14% 25 0% 26 11% 27 7% 28 15% 29 10% 30 12% 31 17% 32 1% 33 18% 34
16% 35 22% 36 15% 37 -22% 38 2% 39 9% 40 9% 41 >100 42 >100
43 ~100 44 >10 45 >100 46 >100 47 >100 48 >100 49
>100 50 >10 51 >100 52 >100 53 >100 54 12.3
TABLE-US-00008 TABLE 2B Aurora-A IC.sub.50 (.mu.M) Cpd IC.sub.50
(avg) 2 10.7 3 >10 4 10.6 5 >10 6 >10 7 >10 8 6.47 9
>1 10 13.3 11 >1 12 ~10 13 5.93 14 >100 15 >100 18
>1 19 9% 20 21% 21 3% 22 15% 23 16% 24 7% 25 18% 26 23% 27 16%
28 25% 29 13% 30 22% 31 22% 32 13% 33 29% 34 29% 35 16% 36 21% 37
3% 38 4% 39 15% 40 15% 41 >10 42 >10 43 >10 44 1.1 45
>100 46 >1 47 >100 48 >100 49 >100 50 >100 51
>100 52 >100 53 >100 54 15.3
Example 3
HER-2 Kinase Assay
[0238] HER-2 kinase was purified at Proqinase (Freiburg, Germany)
from a construct that consisted of a fusion of GST
(Glutathione-S-Transferase), HIS6-Thrombin and the nucleotides
encoding amino acids 679 to 1255 of HER-2.
[0239] A mixture of a 10.times. kinase reaction buffer (600 mM
Hepes at pH 7.5, 30 mM Magnesium Chloride, 0.03 mM Sodium Vanadate
and 500 .mu.g/mL PEG 20,000), DTT (1.2 mM final from a 10 mM
stock), ATP (1 .mu.M from a 10 mM stock), biotinylated polyGluTyr
(1.5 ng/.mu.L final from stock of 1 .mu.g/.mu.L prepared by Upstate
Biotechnologies, Lake Placid, N.Y.), Manganese Chloride (3 mM final
from a 1 M stock), .gamma.-.sup.33P-ATP (10 .mu.Ci/.mu.L stock) and
water (70 .mu.L/well) was added to each well of a Streptavidin
Flashplate (Cat. #SMP103, NEN, Boston, Mass.).
[0240] Test compound stock (1 .mu.L) was added to the appropriate
wells. Diluted GSTHER2 kinase (6.7 ng/.mu.L diluted into 50 mM
Tris-HCl at pH 8.0 and 0.1% bovine serum albumin) (30 .mu.L) was
added (total volume of 200 ng/well) to initiate the reactions.
[0241] The reaction plates were incubated at 30.degree. C. for 1
hr. The reaction was terminated by aspirating the reaction mixture
from the plate wells and washing the wells three times with a
1.times.PBS stop buffer (300 .mu.L) and 100 mM EDTA. After the
final wash, the same stop buffer (200 .mu.L) was again added to the
wells. The plates were then sealed and read on the TopCount
scintillation counter.
[0242] The IC.sub.50 was derived according to the procedure
described in Example 2. The IC.sub.50 results for HER-2 are shown
in Table 3. For those compounds without an IC.sub.50, the percent
inhibition results are shown at a test concentration of 1
.mu.M.
TABLE-US-00009 TABLE 3 HER-2 IC.sub.50 (.mu.M) Cpd IC.sub.50 (avg)
2 3.46 3 4.10 4 5.35 5 >10 6 ~100 7 >100 8 ~100 9 >10 10
7.16 11 2.91 12 5.55 13 >1 14 >100 15 >100 18 1.23 19 49%
20 41% 21 13% 22 48% 23 44% 24 12% 25 3% 26 >1 27 0% 28 35% 29
0% 30 35% 31 35% 32 22% 33 37% 34 >1 35 >1 36 29% 37 11% 38
0% 39 0% 40 0% 41 13.5 42 >1 43 10.6 44 1.1 45 0.0836 46 0.134
47 1.87 48 >10 49 0.836 50 2.20 51 >10 52 >10 53 >100
54 3.37
Example 4
c-Src Kinase Assay
[0243] A mixture of a 10.times. kinase buffer (80 mM MOPS at pH
7.0, 2 mM EDTA and 100 mM Magnesium Chloride), ATP (5 .mu.M final
from a 10 mM stock), a Cdc2 peptide KVEKIGEGTYGVVYK (100 .mu.M
final from a 2.5 mM stock), .gamma.-.sup.33P ATP (10 .mu.Ci/.mu.L
stock) and water (20 .mu.L/well) was added to each well of a
Streptavidin Flashplate.
[0244] Test compound in 100% DMSO (0.5 .mu.L) was added to the
appropriate wells. Diluted c-Src kinase (human) (Upstate
Biotechnology, Lake Placid, N.Y.) (diluted in a buffer consisting
of 20 mM MOPS at pH 7.0, 1 mM EDTA, .beta.-mercaptoethanol (0.1%),
Brij-35 (0.01%), glycerol (5%) and 1 mg/mL bovine serum albumin)
(2.5 .mu.L) was added to the wells to initiate the reactions. The
reaction plates were incubated at 30.degree. C. for 40 min. The
reaction was terminated by the addition of a 3% phosphoric acid
solution (5 .mu.L). The reaction product (10 .mu.L) was spotted
onto a P30 filtermat and washed for 5 minutes in phosphoric acid
(75 mM). The wash sequence was repeated two more times, followed
with one final wash in methanol. The plates were then dried, sealed
and read on the TopCount scintillation counter after adding 30
.mu.L scintillation fluid.
[0245] Percent inhibition was derived according to the procedure
described in Example 2. The IC.sub.50 results for c-Src are shown
in Table 4. For those compounds without an IC.sub.50, the percent
inhibition results are shown at a test concentration of 2
.mu.M.
TABLE-US-00010 TABLE 4 c-Src IC.sub.50 (.mu.M) Cpd IC.sub.50 (avg)
2 3.12 3 30% 4 0.0574 5 0.355 6 0.368 7 -1% 8 -21% 9 11% 10 0.0870
11 0.0712 12 91% 13 78% 14 11% 15 -0.6% 19 90%
Example 5
Lyn Kinase Assay
[0246] A mixture of a 10.times. kinase buffer (500 mM MOPS at pH
7.5, 1 mM EGTA, 1 mM Sodium Vanadate, 1% .beta.-mercaptoethanol and
100 mM Magnesium Acetate), ATP (5 .mu.M final from a 10 mM stock),
polyGluTyr (0.1 mg/mL final from a 1 mg/mL stock), .gamma.-.sup.33P
ATP (10 .mu.Ci/mL stock) and water (20 .mu.L/well) was added to
each well of a Streptavidin Flashplate.
[0247] Test compound in 100% DMSO (0.5 .mu.L) was added to the
appropriate wells. Diluted Lyn kinase (human) (Upstate
Biotechnology, Lake Placid, N.Y.) (diluted in a buffer consisting
of 50 mM Tris at pH 7.5, 0.1 mM EGTA, Sodium Vanadate (0.1 mM),
.beta.-mercaptoethanol (0.1%) and 1 mg/mL bovine serum albumin)
(2.5 .mu.L) was added to the wells to initiate the reactions.
[0248] The reaction plates were incubated at 30.degree. C. for 40
min. The reaction was terminated by the addition of a 3% phosphoric
acid solution (5 .mu.L). The reaction product (10 .mu.L) was
spotted onto a P30 filtermat and washed for 5 minutes in phosphoric
acid (75 mM). The wash sequence was repeated two more times,
followed with one final wash in methanol. The plates were then
dried, sealed and read on the TopCount scintillation counter after
adding 30 .mu.L scintillation fluid.
[0249] Percent inhibition was derived according to the procedure
described in Example 2. The IC.sub.50 results for Lyn are shown in
Table 5. For those compounds without an IC.sub.50, the percent
inhibition results are shown at a test concentration of 2
.mu.M.
TABLE-US-00011 TABLE 5 Lyn IC.sub.50 (.mu.M) Cpd IC.sub.50 (avg) 2
2.99 3 2% 4 0.477 5 83% 6 75% 7 43% 8 40% 9 58% 10 0.268 11 0.113
12 93% 13 76% 18 86%
Example 6
c-Abl Kinase Assay
[0250] A mixture of a 10.times. kinase buffer (80 mM MOPS at pH
7.0, 2 mM EDTA and 100 mM Magnesium Acetate), ATP (5 .mu.M final
from a 10 mM stock), a peptide EAIYAAPFAKKK (50 .mu.M final from a
0.5 mM stock), .gamma.-.sup.33P ATP (10 .mu.Ci/.mu.L stock) and
water is added to each well (20 .mu.L/well) of a Streptavidin
Flashplate.
[0251] Test compound in 100% DMSO (0.5 .mu.L) is added to the
appropriate wells. Diluted c-Abl kinase (human) (Upstate
Biotechnology, Lake Placid, N.Y.) (diluted in a buffer consisting
of 20 mM MOPS at pH 7.0, 1 mM EDTA, .beta.-mercaptoethanol (0.1%),
Brij-35 (0.01%), glycerol (5%) and 1 mg/ml bovine serum albumin)
(2.5 .mu.L) is added to the wells to initiate the reactions.
[0252] The reaction plates are incubated at 30.degree. C. for 40
min. The reaction is terminated by the addition of a 3% phosphoric
acid solution (5 .mu.L). The reaction product (10 .mu.L) is spotted
onto a P30 filtermat and is washed for 5 minutes in phosphoric acid
(75 mM). The wash sequence is repeated two more times and is
followed with one final wash in methanol. The plates are then
dried, sealed and read on the TopCount scintillation counter after
30 .mu.L scintillation fluid is added. The IC.sub.50 is derived
according to the procedure described in Example 2.
Example 7
Cell Proliferation Inhibition Assay
[0253] The ability of a test compound to inhibit unregulated cell
proliferation was determined by measuring incorporation of
.sup.14C-labelled thymidine into newly synthesized DNA within cell
lines derived from carcinomas originating from several tissues.
Accordingly, the anti-proliferative effect of a compound on cells
with a variety of phenotypes may be determined.
[0254] Carcinoma cell lines include those such as HeLa cervical
adenocarcinoma (American Type Culture Collection (ATCC), Virginia,
Cat. #CCL-2), A375 malignant melanoma (ATCC CRL-1619), SK-OV-3
ovarian adenocarcinoma (ATCC HTB-77), HCT-116 colon carcinoma
(CCL-247), PC-3 prostate adenocarcinoma (ATCC CRL-1435), and
MDA-MB-231 (Xenogen Corp.)
[0255] The carcinoma cells were trypsinized and counted. The cells
(3000-8000 count) were added to each well of a 96-well CytoStar
tissue culture treated scintillating microplate (Amersham
#RPNQ0160) in complete medium (100 .mu.L) and the plate was then
incubated in complete medium for 24 hrs at 37.degree. C. in an
inert atmosphere containing 5% CO.sub.2. Test compound (1 .mu.L) in
100% DMSO was added to the plate test-wells with DMSO only added to
control-wells. The plate was incubated in complete medium for a
second 24 hr period at 37.degree. C. in an atmosphere containing 5%
CO.sub.2.
[0256] An aliquot of a solution of Methyl .sup.14C-thymidine (56
mC/mmol) (NEN #NEC568 or Amersham #CFA532) and complete medium (20
uL to provide 0.2 .mu.Ci/well) was then added to each well and the
plate was incubated for a third 24 hr period at 37.degree. C. in an
atmosphere containing 5% CO.sub.2. The plate contents were then
discarded, the plate was washed twice with PBS (200 .mu.L) and then
PBS (200 .mu.L) was added to each well. The plate was sealed and
the degree of methyl .sup.14C-thymidine incorporation was
quantified on a Packard Top Count.
TABLE-US-00012 TABLE 6 Cell Proliferation IC.sub.50 (avg, .mu.M)
Cpd A375 HCT116 HeLa 41 >100 >10 >100 42 >100 >100
>100 45 >10 >10 >100 46 >10 >10 >10 48 8.29
15.8 >10 49 1.38 0.985 2.19
Example 8
In Vivo Models
Inhibition of Tumor Growth
[0257] The ability of a test compound to inhibit unregulated growth
of human tumor cells in vivo may be evaluated by implanting human
tumor cells into the hindflank of athymic mice, administering a
test compound and then quantifying any change in tumor size.
[0258] Human epidermoid A431 carcinoma cells (10.sup.6 count) are
implanted subcutaneously into the hindflank of female athymic mice
(Charles River) and allowed to grow for 6-10 days. After a
measurable tumor is established (as determined by baseline caliper
measurement), the animal is administered an oral dose of the test
compound (in 10% solutol) daily for a period of 30 days. Tumor size
is measured every five days and the degree of inhibition is
determined by comparing drug-treated animals to vehicle-treated
animals.
[0259] Variations of this method are intended to include
intraperitoneal injection or intravenous infusion as the route of
administration and administration of the test compound either alone
or in a combination therapy.
[0260] While the foregoing specification teaches the principles of
the present invention, with examples provided for the purpose of
illustration, it will be understood that the practice of the
invention encompasses all of the usual variations, adaptations and
modifications as come within the scope of the following claims and
their equivalents.
[0261] Throughout this application, various publications are cited.
The disclosure of these publications is hereby incorporated by
reference into this application to describe more fully the state of
the art to which this invention pertains.
* * * * *