U.S. patent application number 11/285516 was filed with the patent office on 2006-06-08 for bicyclic inhibitors of rho kinase.
Invention is credited to Upul Bandarage, Jingrong Cao, Cornelia Forster, Huai Gao, Jeremy Green.
Application Number | 20060122185 11/285516 |
Document ID | / |
Family ID | 35953926 |
Filed Date | 2006-06-08 |
United States Patent
Application |
20060122185 |
Kind Code |
A1 |
Green; Jeremy ; et
al. |
June 8, 2006 |
Bicyclic inhibitors of Rho kinase
Abstract
The present invention relates to compounds useful as inhibitors
of protein kinases, particularly of ROCK. The invention also
provides pharmaceutically acceptable compositions comprising said
compounds and methods of using the compositions in the treatment of
various disease, conditions, or disorders.
Inventors: |
Green; Jeremy; (Waltham,
MA) ; Cao; Jingrong; (Newton, MA) ; Bandarage;
Upul; (Lexington, MA) ; Gao; Huai; (Arlington,
MA) ; Forster; Cornelia; (Pelham, NH) |
Correspondence
Address: |
VERTEX PHARMACEUTICALS INC.
130 WAVERLY STREET
CAMBRIDGE
MA
02139-4242
US
|
Family ID: |
35953926 |
Appl. No.: |
11/285516 |
Filed: |
November 22, 2005 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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60630115 |
Nov 22, 2004 |
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Current U.S.
Class: |
514/248 ;
514/249; 514/262.1; 514/303; 544/236; 544/262; 544/350;
546/117 |
Current CPC
Class: |
A61P 37/04 20180101;
C07D 471/04 20130101; A61P 25/28 20180101; A61P 37/00 20180101;
A61P 25/00 20180101; A61P 35/02 20180101; A61P 27/06 20180101; A61P
11/06 20180101; A61P 15/10 20180101; A61P 35/00 20180101; A61P
19/08 20180101; A61P 37/08 20180101; C07D 487/04 20130101; A61P
25/18 20180101; A61P 31/12 20180101; A61P 37/06 20180101; A61P 1/16
20180101; A61P 43/00 20180101; A61P 31/00 20180101; A61P 3/10
20180101; A61P 19/10 20180101; A61P 9/12 20180101; A61P 9/00
20180101; A61P 19/00 20180101; A61P 29/00 20180101; A61P 9/10
20180101; A61P 19/02 20180101 |
Class at
Publication: |
514/248 ;
514/249; 514/262.1; 514/303; 544/236; 544/262; 544/350;
546/117 |
International
Class: |
A61K 31/519 20060101
A61K031/519; A61K 31/503 20060101 A61K031/503; A61K 31/498 20060101
A61K031/498; A61K 31/4745 20060101 A61K031/4745; C07D 471/02
20060101 C07D471/02; C07D 487/02 20060101 C07D487/02 |
Claims
1. A compound of formula I: ##STR57## or a pharmaceutically
acceptable salt thereof, wherein: wherein ##STR58## Z.sub.1,
Z.sub.2, Z.sub.3 and Z.sub.4 are each independently selected from N
or CR.sup.1, wherein at least one of Z.sub.1, Z.sub.2, or Z.sub.4
is N; each R.sup.1 is independently selected from H, halogen, --CN,
--NO.sub.2, or --V.sub.mR'; G is --NR.sup.2-- or --CO--; Q.sup.1 is
--CO--, --SO.sub.2--, --NR.sup.2, --NR.sup.2CO--, --CONR.sup.2--,
--SO.sub.2NR.sup.2--, or is a bond; R.sup.2 is --U.sub.nR'';
R.sup.3 is Q.sup.2-Ar.sup.1, or when G is --NR.sup.2, R.sup.2 and
Q.sup.1-R.sup.3, taken together with the nitrogen atom, may form
the cyclic group: ##STR59## where s is 1 or 2, Z is CH or N;
wherein each occurrence of Y is independently --CO--, --CS--,
--SO.sub.2--, --O--, --S--, --NR.sup.5--, or --C(R.sup.5).sub.2--,
and R.sup.5 is U.sub.nR'; X.sub.1 and X.sub.2 are each
independently selected from CR.sup.4 or N; each occurrence of
R.sup.4 is independently selected from halogen, CN, NO.sub.2, or
V.sub.mR; each occurrence of U or V is independently selected from
an optionally substituted C.sub.1-6 alkylidene chain, wherein up to
two methylene units of the chain are optionally and independently
replaced by --NR--, --S--, --O--, --CS--, --CO.sub.2--, --OCO--,
--CO--, --COCO--, --CONR--, --NRCO--, --NRCO.sub.2--,
--SO.sub.2NR--, --NRSO.sub.2--, --CONRNR--, --NRCONR--, --OCONR--,
--NRNR--, --NRSO.sub.2NR--, --SO--, --SO.sub.2--, --PO--,
--PO.sub.2--, or --POR--; m and n are each independently 0 or 1;
each occurrence of R is independently selected from hydrogen or a
C.sub.1-6 aliphatic group, wherein said aliphatic group is
optionally substituted with up to five occurrences of J.sup.R; each
occurrence of R' is independently selected from hydrogen, a
C.sub.1-6 aliphatic group, a 3-8-membered saturated, partially
unsaturated, or fully unsaturated monocyclic ring having 0-3
heteroatoms independently selected from nitrogen, oxygen, or
sulfur, or an 8-12 membered saturated, partially unsaturated, or
fully unsaturated bicyclic ring system having 0-5 heteroatoms
independently selected from nitrogen, oxygen, or sulfur, wherein
said aliphatic group, monocyclic ring or bicyclic ring is
optionally substituted with up to five occurrences of J.sup.R'; R''
is selected from hydrogen, a C.sub.1-6 aliphatic group, a
3-8-membered saturated, partially unsaturated, or fully unsaturated
monocyclic ring having 0-3 heteroatoms independently selected from
nitrogen, oxygen, or sulfur, or an 8-12 membered saturated,
partially unsaturated, or fully unsaturated bicyclic ring system
having 0-5 heteroatoms independently selected from nitrogen,
oxygen, or sulfur, wherein said aliphatic group, monocyclic ring or
bicyclic ring is optionally substituted with up to five occurrences
of J.sup.R''; or two occurrences of R, R' and R'', in any
combination thereof, are taken together with the atom(s) to which
they are bound to form a 3-12 membered saturated, partially
unsaturated, or fully unsaturated monocyclic or bicyclic ring
having 0-4 heteroatoms independently selected from nitrogen,
oxygen, or sulfur, wherein said monocyclic or bicyclic ring is
optionally substituted with J.sup.R; each occurrence of J.sup.R,
J.sup.R' and J.sup.R'' is independently selected from halogen, L,
-(L.sub.p)-R.sup.J, -(L.sub.p)-N(R.sup.J).sub.2,
-(L.sub.p)-SR.sup.J, -(L.sub.p)-OR.sup.J, -(L.sub.p)-(C.sub.3-10
cycloaliphatic), -(L.sub.p)-(C.sub.6-10 aryl), -(L.sub.p)-(5-10
membered heteroaryl), -(L.sub.p)-(5-10 membered heterocyclyl), oxo,
C.sub.1-4haloalkoxy, C.sub.1-4haloalkyl, -(L.sub.p)-NO.sub.2,
-(L.sub.p)-CN, -(L.sub.p)-OH, -(L.sub.p)-CF.sub.3,
--CO.sub.2R.sup.J, --CO.sub.2H, --COR.sup.J, --COH, --OC(O)R.sup.J
or --NC(O)R.sup.J; or any two J.sup.R, J.sup.R' or J.sup.R''
groups, on the same substituent or different substituents, together
with the atom(s) to which each J.sup.R, J.sup.R' or J.sup.R'' group
is bound, form a 5-7 membered saturated, unsaturated, or partially
saturated ring; R.sup.J is H or C.sub.1-6 aliphatic; or two R.sup.J
groups or an R.sup.J group and an R, R' or R'' group, together with
the atom to which they are attached, optionally form a 3-6 membered
cycloaliphatic or heterocyclyl, wherein said aliphatic,
cycloaliphatic or heterocyclyl is optionally substituted with R*,
--OR*, --SR*, --NO.sub.2, --CF.sub.3, --CN, --CO.sub.2R*, --COR*,
OCOR* or NHCOR*, wherein R* is H or an unsubstituted C.sub.1-6
aliphatic; L is a C.sub.1-6 aliphatic wherein up to three methylene
units are replaced by --NH--, --NR.sup.L--, --O--, --S--,
--CO.sub.2--, --OC(O)--, --C(O)CO--, --C(O)--, --C(O)NH--,
--C(O)NR.sup.6--, --C(.dbd.N--CN), --NHCO--, --NR.sup.LCO--,
--NHC(O)O--, --NR.sup.LC(O)O--, --SO.sub.2NH--,
--SO.sub.2NR.sup.L--, --NHSO.sub.2--, --NR.sup.LSO.sub.2--,
--NHC(O)NH--, --NR.sup.LC(O)NH--, --NHC(O)NR.sup.L--,
--NR.sup.LC(O)NR.sup.L, --OC(O)NH--, --OC(O)NR.sup.L--,
--NHSO.sub.2NH--, --NR.sup.LSO.sub.2NH--, --NHSO.sub.2NR.sup.L--,
--NR.sup.LSO.sub.2NR.sup.L--, --SO-- or --SO.sub.2--; R.sup.L is
selected from C.sub.1-6 aliphatic, C.sub.3-10 cycloaliphatic,
C.sub.6-10 aryl, 5-10 membered heteroaryl or 5-10 membered
heterocyclyl; or two R.sup.L groups, on the same substituent or
different substituents, together with the atom(s) to which each
R.sup.L group is bound, form a 3-8 membered heterocyclyl; each p is
independently 0 or 1; Q.sup.2 and Q.sup.3 are each independently
selected from a bond or a C.sub.1-6 alkylidene chain, wherein up to
two methylene units of the chain are each optionally and
independently replaced by --NR'--, --S--, --O--, --CS--,
--CO.sub.2--, --OCO--, --CO--, --COCO--, --CONR'--, --NR'CO--,
--NR'CO.sub.2--, --SO.sub.2NR'--, --NR'SO.sub.2--, --CONR'NR'--,
--NR'CONR'--, --OCONR'--, --NR'NR'--, --NR'SO.sub.2NR'--, --SO--,
--SO.sub.2--, --PO--, --PO.sub.2--, or --POR'--; and wherein any
carbon atom in the one or more methylene units is optionally
substituted with one or two occurrences of R.sup.6, wherein each
occurrence of R.sup.6 is independently halogen, --CN, --NO.sub.2,
or --U.sub.nR', or two occurrences of R.sup.6, or R' and R.sup.6,
taken together with the atoms to which they are bound, form an
optionally substituted 3-6-membered cycloalkyl, heterocyclyl, aryl
or heteroaryl ring; and Ar.sup.1 and Ar.sup.2 are each
independently selected from a C.sub.1-6 aliphatic, a 3-8 membered
saturated, partially unsaturated, or fully unsaturated monocyclic
ring having 0-3 heteroatoms independently selected from nitrogen,
oxygen, or sulfur, or an 8-12 membered saturated, partially
unsaturated, or fully unsaturated bicyclic ring system having 0-5
heteroatoms independently selected from nitrogen, oxygen, or
sulfur; wherein Ar.sup.1 and Ar.sup.2 are each optionally
substituted with 0-5 independent occurrences of TR.sup.7; wherein T
is a bond or is a C.sub.1-C.sub.6 alkylidene chain wherein up to
two methylene units of T are optionally and independently replaced
by --NR--, --S--, --O--, --CS--, --CO.sub.2--, --OCO--, --CO--,
--COCO--, --CONR--, --NRCO--, --NRCO.sub.2--, --SO.sub.2NR--,
--NRSO.sub.2--, --CONRNR--, --NRCONR--, --OCONR--, --NRNR--,
--NRSO.sub.2NR--, --SO--, --SO.sub.2--, --PO--, --PO.sub.2--, or
--POR--; and each occurrence of R.sup.7 is independently selected
from --R', halogen, --NO.sub.2, --CN or .dbd.O.
2. The compound according to claim 1, having one of formulae
II-VII: ##STR60## ##STR61## wherein R.sup.1Z1, R.sup.1Z2, R.sup.1Z3
and R.sup.1Z4 are each independently selected from H, halogen,
--CN, --NO.sub.2, or --V.sub.mR'.
3-4. (canceled)
5. The compound according to claim 1, wherein X.sub.1 is CR.sup.4
and X.sub.2 is N or CR.sup.4.
6. (canceled)
7. The compound according to claim 1, wherein each R.sup.1 is
independently selected from H, halogen or C.sub.1-3 aliphatic.
8-13. (canceled)
14. The compound according to claim 1, wherein Q.sup.1 is --CO--,
--SO.sub.2--, --NR.sup.2, --NR.sup.2CO--, --CONR.sup.2--,
--SO.sub.2NR.sup.2.
15. The compound according to claim 14, wherein G is --NR.sup.2 and
Q.sup.1 is --CO--, or G is --CO-- and Q.sup.1 is --NR.sup.2--.
16. The compound according to claim 15, wherein R.sup.2 is H,
--C.sub.1-4 aliphatic, -cyclopropyl, (CH.sub.2).sub.1-3OH or
##STR62##
17. (canceled)
18. The compound according to claim 1, wherein R.sup.3 is
Q.sup.2-Ar.sup.1.
19. The compound according to claim 18, wherein Q.sup.2 is
--(CHR.sup.6).sub.q--, --(CHR.sup.6).sub.qO--,
--(CHR.sup.6).sub.qS--, --(CHR.sup.6).sub.qS(O).sub.2--,
--(CHR.sup.6).sub.qS(O)--, --(CHR.sup.6).sub.qNR--, or
--(CHR.sup.6).sub.qC(O)--, wherein q is 0, 1, 2, or 3, and each
R.sup.6 is R', --N(R)(R'), --(CH.sub.2).sub.1-4N(R)(R'),
--(CH.sub.2).sub.1-4C(CH.sub.3).sub.2N(R)(R'),
--(CH.sub.2).sub.1-4CH(CH.sub.3)N(R)(R'), --OR',
--(CH.sub.2).sub.1-4OR', --NR(CH.sub.2).sub.1-4N(R)(R'),
--NR(CH.sub.2).sub.1-4SO.sub.2R', --NR(CH.sub.2).sub.1-4COOR', or
--NR(CH.sub.2).sub.1-4COR', or two occurrences of R.sup.6, taken
together with the atoms to which they are bound, form an optionally
substituted 3-6-membered saturated, partially unsaturated, or fully
unsaturated ring.
20-21. (canceled)
22. The compound according to claim 18, wherein Ar.sup.1 is a
C.sub.3-6 aliphatic, a 5-8 membered saturated, partially
unsaturated, or fully unsaturated monocyclic ring having 0-3
heteroatoms independently selected from nitrogen, oxygen, or
sulfur, or an 8-12 membered saturated, partially unsaturated, or
fully unsaturated bicyclic ring system having 0-5 heteroatoms
independently selected from nitrogen, oxygen, or sulfur; wherein
Ar.sup.1 is optionally substituted with 0-5 independent occurrences
of TR.sup.7.
23. The compound according to claim 22, wherein Ar.sup.1 is
##STR63## ##STR64## ##STR65## ##STR66## wherein t is 0, 1, 2, 3, 4,
or 5, and wherein any Ar.sup.1 is bonded to Q.sup.2 through any
substitutable nitrogen or carbon atom, and wherein one or more
hydrogen atoms on any substitutable nitrogen or, carbon atom is
substituted with one or more independent occurrences of
TR.sup.7.
24-26. (canceled)
27. The compound according to claim 22, wherein TR.sup.7 is
selected from --F, --Cl, --CN, --NH.sub.2, --CH.sub.3,
--CH.sub.2CH.sub.3, --CH(CH.sub.3).sub.2, --OR.sup.x, --OCF.sub.3,
--NR.sup.xSO.sub.2R.sup.x, --NR.sup.xSO.sub.2N(R.sup.x).sub.2,
--COOC(CH.sub.3).sub.3, --OSO.sub.2CH.sub.3, --OH,
--SO.sub.2N(R.sup.x).sub.2, --SO.sub.2N(R.sup.x).sub.2,
--SO.sub.2R.sup.x, -pyrollidinone, tetrahydrofuran or
-D-(CH.sub.2).sub.p--Y, wherein R.sup.x is a H or a C.sub.1-4
alkyl, D is --SO.sub.2--, --SO.sub.2NH--, --NHSO.sub.2-- or --O--,
p is 0-3, and Y is selected from: ##STR67## wherein R.sup.y is H or
C.sub.1-3 alkyl, and wherein one or more carbon atoms of Y is
optionally substituted with .dbd.O.
28-31. (canceled)
32. The compound according to claim 1, wherein said compound has a
structure depicted in Table 1.
33. A composition comprising an effective amount of compound
according to claim 1, and a pharmaceutically acceptable carrier,
adjuvant, or vehicle.
34. The composition of claim 33, additionally comprising a
therapeutic agent selected from a chemotherapeutic or
anti-proliferative agent, an anti-inflammatory agent, an
immunomodulatory or immunosuppressive agent, a neurotrophic factor,
an agent for treating cardiovascular disease, an agent for treating
destructive bone disorders, an agent for treating liver disease, an
anti-viral agent, an agent for treating blood disorders, an agent
for treating diabetes, or an agent for treating immunodeficiency
disorders.
35. A method of inhibiting ROCK kinase activity in a biological
sample; which method contacting said biological sample with a
compound of claim 1 or a composition comprising said compound.
36. A method of treating or lessening the severity of a disease
condition or disorder selected from a proliferative disorder, a
cardiac disorder, a neurodegenerative disorder, a psychotic
disorder, an autoimmune disorder, a condition associated with organ
transplant, an inflammatory disorder, an immunologically mediated
disorder, a viral disease, or a bone disorder, comprising the step
of administering to said patient a compound according to claim 1 or
a composition comprising said compound.
37. The method of claim 36, comprising the additional step of
administering to said patient an additional therapeutic agent
selected from a chemotherapeutic or anti-proliferative agent, an
anti-inflammatory agent, an immunomodulatory or immunosuppressive
agent, a neurotrophic factor, an anti-psychotic agent, an agent for
treating cardiovascular disease, an agent for treating destructive
bone disorders, an agent for treating liver disease, an anti-viral
agent, an agent for treating blood disorders, an agent for treating
diabetes, or an agent for treating immunodeficiency disorders,
wherein: said additional therapeutic agent is appropriate for the
disease being treated; and said additional therapeutic agent is
administered together with said composition as a single dosage form
or separately from said composition as part of a multiple dosage
form.
38-40. (canceled)
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional
Application No. 60/630,115, filed Nov. 22, 2005, which is herein
incorporated by reference.
TECHNICAL FIELD OF INVENTION
[0002] The present invention relates to compounds useful as
inhibitors of protein kinases. The invention also provides
pharmaceutically acceptable compositions comprising the compounds
of the invention and methods of using the compositions in the
treatment of various disorders.
BACKGROUND OF THE INVENTION
[0003] Protein kinases constitute a large family of structurally
related enzymes that are responsible for the control of a variety
of signal transduction processes within the cell. Many diseases are
associated with abnormal cellular responses triggered by protein
kinase-mediated events. Accordingly, there has been a substantial
effort in medicinal chemistry to find protein kinase inhibitors
that are effective as therapeutic agents.
[0004] One kinase family of interest is Rho-associated coiled-coil
forming protein serine/threonine kinase (ROCK), which is believed
to be an effector of Ras-related small GTPase Rho. The ROCK family
includes p160ROCK (ROCK-1) and ROK.alpha./Rho-kinase/ROCK-II,
protein kinase PKN, and citron and citron kinase. The ROCK family
of kinases have been shown to be involved in a variety of functions
including Rho-induced formation of actin stress fibers and focal
adhesions and in downregulation of myosin phosphatase, platelet
activation, aortic smooth muscle contraction by various stimuli,
thrombin-induced responses of aortic smooth muscle cells,
hypertrophy of cardiomyocytes, bronchial smooth muscle contraction,
smooth muscle contraction and cytoskeletal reorganization of
non-muscle cells, activation of volume-regulated anion channels,
neurite retraction, neutrophil chemotaxis, wound healing, tumor
invasion and cell transformation. More specifically, ROCK has been
implicated in various diseases and disorders including
hypertension, cerebral vasospasm, coronary vasospasm, bronchial
asthma, preterm labor, erectile dysfunction, glaucoma, vascular
smooth muscle cell proliferation, myocardial hypertrophy,
malignoma, ischemia/reperfusion-induced injury, endothelial
dysfunction, Crohn's Disease and colitis, neurite outgrowth,
Raynaud's Disease, angina, Alzheimer's disease, benign prostatic
hyperplasia and atherosclerosis. Accordingly, the development of
inhibitors of ROCK kinase would be useful as therapeutic agents for
the treatment of disorders implicated in the ROCK kinase
pathway.
[0005] Accordingly, there is a great need to develop inhibitors of
ROCK that would be useful in treating various diseases or
conditions associated with ROCK, particularly given the inadequate
treatments currently available for the majority of these
disorders.
SUMMARY OF THE INVENTION
[0006] It has now been found that compounds of this invention, and
pharmaceutically acceptable compositions thereof, are effective as
inhibitors of ROCK. These compounds have the general formula I:
##STR1##
[0007] or a pharmaceutically acceptable derivative thereof, wherein
ring B, Z.sup.1, Z.sup.2, Z.sup.3, Z.sup.4, R.sup.3, G and Q.sup.1
are as defined below.
[0008] These compounds, and pharmaceutically acceptable
compositions thereof, are useful for treating or lessening the
severity of a variety of disorders, including, without limitation,
hypertension, cerebral vasospasm, coronary vasospasm, bronchial
asthma, preterm labor, erectile dysfunction, glaucoma, vascular
smooth muscle cell proliferation, myocardial hypertrophy,
malignoma, ischemia/reperfusion-induced injury, endothelial
dysfunction, Crohn's Disease and colitis, neurite outgrowth,
Raynaud's Disease, angina, Alzheimer's disease, benign prostatic
hyperplasia and atherosclerosis.
DETAILED DESCRIPTION OF THE INVENTION
[0009] Definitions
[0010] As used herein, the following definitions shall apply unless
otherwise indicated. For purposes of this invention, the chemical
elements are identified in accordance with the Periodic Table of
the Elements, CAS version, and the Handbook of Chemistry and
Physics, 75.sup.th Ed. 1994. Additionally, general principles of
organic chemistry are described in "Organic Chemistry", Thomas
Sorrell, University Science Books, Sausalito: 1999, and "March's
Advanced Organic Chemistry", 5.sup.th Ed., Smith, M. B. and March,
J., eds. John Wiley & Sons, New York: 2001, the entire contents
of which are hereby incorporated by reference.
[0011] As described herein, compounds of the invention may
optionally be substituted with one or more substituents, such as
are illustrated generally above, or as exemplified by particular
classes, subclasses, and species of the invention. It will be
appreciated that the phrase "optionally substituted" is used
interchangeably with the phrase "substituted or unsubstituted." In
general, the term "substituted", whether preceded by the term
"optionally" or not, refers to the replacement of one or more
hydrogen radicals in a given structure with the radical of a
specified substituent. Unless otherwise indicated, an optionally
substituted group may have a substituent at each substitutable
position of the group. When more than one position in a given
structure can be substituted with more than one substituent
selected from a specified group, the substituent may be either the
same or different at each position.
[0012] As described herein, when the term "optionally substituted"
precedes a list, said term refers to all of the subsequent
substitutable groups in that list. If a substituent radical or
structure is not identified or defined as "optionally substituted",
the substituent radical or structure is unsubstituted. For example,
if X is halogen; optionally substituted C.sub.1-3alkyl or phenyl; X
may be either optionally substituted alkyl or optionally
substituted phenyl. Likewise, if the term "optionally substituted"
follows a list, said term also refers to all of the substitutable
groups in the prior list unless otherwise indicated. For example:
if X is halogen, C.sub.1-3alkyl or phenyl wherein X is optionally
substituted by J.sup.X, then both C.sub.1-3alkyl and phenyl may be
optionally substituted by J.sup.X. As is apparent to one having
ordinary skill in the art, groups such as H, halogen, NO.sub.2, CN,
NH.sub.2, OH, or OCF.sub.3 would not be included because they are
not substitutable groups.
[0013] Combinations of substituents envisioned by this invention
are preferably those that result in the formation of stable or
chemically feasible compounds. The term "stable", as used herein,
refers to compounds that are not substantially altered when
subjected to conditions to allow for their production, detection,
and, preferably, their recovery, purification, and use for one or
more of the purposes disclosed herein. In some embodiments, a
stable compound or chemically feasible compound is one that is not
substantially altered when kept at a temperature of 40.degree. C.
or less, in the absence of moisture or other chemically reactive
conditions, for at least a week.
[0014] The term "aliphatic" or "aliphatic group", as used herein,
means a straight-chain (i.e., unbranched) or branched, substituted
or unsubstituted hydrocarbon chain that is completely saturated or
that contains one or more units of unsaturation. Unless otherwise
specified, aliphatic groups contain 1-20 aliphatic carbon atoms. In
some embodiments, aliphatic groups contain 1-10 aliphatic carbon
atoms. In other embodiments, aliphatic groups contain 1-8 aliphatic
carbon atoms. In still other embodiments, aliphatic groups contain
1-6 aliphatic carbon atoms, and In yet other embodiments, aliphatic
groups contain 1-4 aliphatic carbon atoms. Suitable aliphatic
groups include, but are not limited to, linear or branched,
substituted or unsubstituted alkyl, alkenyl, or alkynyl groups.
Further examples of aliphatic groups include methyl, ethyl, propyl,
butyl, isopropyl, isobutyl, vinyl, and sec-butyl.
[0015] The term "cycloaliphatic" (or "carbocycle" or "cycloalkyl")
refers to a monocyclic C.sub.3-C.sub.8 hydrocarbon or bicyclic
C.sub.8-C.sub.12 hydrocarbon that is completely saturated or that
contains one or more units of unsaturation, but which is not
aromatic, that has a single point of attachment to the rest of the
molecule, and wherein any individual ring in said bicyclic ring
system has 3-7 members. Suitable cycloaliphatic groups include, but
are not limited to, cycloalkyl, cycloalkenyl, and cycloalkynyl.
Further examples of aliphatic groups include cyclopentyl,
cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and
cycloheptenyl.
[0016] The term "heterocycle", "heterocyclyl",
"heterocycloaliphatic", or "heterocyclic" as used herein refers to
a monocyclic, bicyclic, or tricyclic ring system in which one or
more ring members are an independently selected heteroatom and that
is completely saturated or that contains one or more units of
unsaturation, but which is not aromatic, that has a single point of
attachment to the rest of the molecule. In some embodiments, the
"heterocycle", "heterocyclyl", "heterocycloaliphatic", or
"heterocyclic" group has three to fourteen ring members in which
one or more ring members is a heteroatom independently selected
from oxygen, sulfur, nitrogen, or phosphorus, and each ring in the
system contains 3 to 7 ring members.
[0017] Examples of heterocyclic rings include, but are not limited
to, the following monocycles: 2-tetrahydrofuranyl,
3-tetrahydrofuranyl, 2-tetrahydrothiophenyl,
3-tetrahydrothiophenyl, 2-morpholino, 3-morpholino, 4-morpholino,
2-thiomorpholino, 3-thiomorpholino, 4-thiomorpholino,
1-pyrrolidinyl, 2-pyrrolidinyl, 3-pyrrolidinyl,
1-tetrahydropiperazinyl, 2-tetrahydropiperazinyl,
3-tetrahydropiperazinyl, 1-piperidinyl, 2-piperidinyl,
3-piperidinyl, 1-pyrazolinyl, 3-pyrazolinyl, 4-pyrazolinyl,
5-pyrazolinyl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl,
4-piperidinyl, 2-thiazolidinyl, 3-thiazolidinyl, 4-thiazolidinyl,
1-imidazolidinyl, 2-imidazolidinyl, 4-imidazolidinyl,
5-imidazolidinyl; and the following bicycles:
3-1H-benzimidazol-2-one, 3-(1-alkyl)-benzimidazol-2-one, indolinyl,
tetrahydroquinolinyl, tetrahydroisoquinolinyl, benzothiolane,
benzodithiane, and 1,3-dihydro-imidazol-2-one.
[0018] The term "heteroatom" means one or more of oxygen, sulfur,
nitrogen, phosphorus, or silicon, including any oxidized form of
nitrogen, sulfur, phosphorus, or silicon, the quaternized form of
any basic nitrogen, or a substitutable nitrogen of a heterocyclic
ring, for example N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in
pyrrolidinyl) or NR.sup.+ (as in N-substituted pyrrolidinyl).
[0019] The term "unsaturated", as used herein, means that a moiety
has one or more units of unsaturation.
[0020] The term "alkoxy", or "thioalkyl", as used herein, refers to
an alkyl group, as previously defined, attached to the principal
carbon chain through an oxygen ("alkoxy") or sulfur ("thioalkyl")
atom.
[0021] The terms "haloalkyl", "haloalkenyl" and "haloalkoxy" means
alkyl, alkenyl or alkoxy, as the case may be, substituted with one
or more halogen atoms. The term "halogen" means F, Cl, Br, or
I.
[0022] The term "aryl" used alone or as part of a larger moiety as
in "aralkyl", "aralkoxy", or "aryloxyalkyl", refers to monocyclic,
bicyclic, and tricyclic carbocyclic ring systems having a total of
six to fourteen ring members, wherein at least one ring in the
system is aromatic, wherein each ring in the system contains 3 to 7
ring members and that has a single point of attachment to the rest
of the molecule. The term "aryl" may be used interchangeably with
the term "aryl ring". Examples of aryl rings would include phenyl,
naphthyl, and anthracene.
[0023] The term "heteroaryl", used alone or as part of a larger
moiety as in "heteroaralkyl" or "heteroarylalkoxy", refers to
monocyclic, bicyclic, and tricyclic ring systems having a total of
five to fourteen ring members, wherein at least one ring in the
system is aromatic, at least one ring in the system contains one or
more heteroatoms, wherein each ring in the system contains 3 to 7
ring members and that has a single point of attachment to the rest
of the molecule. The term "heteroaryl" may be used interchangeably
with the term "heteroaryl ring" or the term "heteroaromatic".
[0024] Further examples of heteroaryl rings include the following
monocycles: 2-furanyl, 3-furanyl, N-imidazolyl, 2-imidazolyl,
4-imidazolyl, 5-imidazolyl, 3-isoxazolyl, 4-isoxazolyl,
5-isoxazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, N-pyrrolyl,
2-pyrrolyl, 3-pyrrolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl,
2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, pyridazinyl (e.g.,
3-pyridazinyl), 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, tetrazolyl
(e.g., 5-tetrazolyl), triazolyl (e.g., 2-triazolyl and
5-triazolyl), 2-thienyl, 3-thienyl, pyrazolyl (e.g., 2-pyrazolyl),
isothiazolyl, 1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl,
1,2,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,3-thiadiazolyl,
1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, pyrazinyl, 1,3,5-triazinyl,
and the following bicycles: benzimidazolyl, benzofuryl,
benzothiophenyl, indolyl (e.g., 2-indolyl), purinyl, quinolinyl
(e.g., 2-quinolinyl, 3-quinolinyl, 4-quinolinyl), and isoquinolinyl
(e.g., 1-isoquinolinyl, 3-isoquinolinyl, or 4-isoquinolinyl).
[0025] In some embodiments, an aryl (including aralkyl, aralkoxy,
aryloxyalkyl and the like) or heteroaryl (including heteroaralkyl
and heteroarylalkoxy and the like) group may contain one or more
substituents. Suitable substituents on the unsaturated carbon atom
of an aryl or heteroaryl group are selected from those listed in
the definition of J.sup.Q, J.sup.R, J.sup.V, J.sup.U and J.sup.X
below. Other suitable substituents include: halogen; --R.sup.o;
--OR.sup.o; --SR.sup.o; 1,2-methylenedioxy; 1,2-ethylenedioxy;
phenyl (Ph) optionally substituted with R.sup.o; --O(Ph) optionally
substituted with R.sup.o; --(CH.sub.2).sub.1-2(Ph), optionally
substituted with R.sup.o; --CH.dbd.CH(Ph), optionally substituted
with R.sup.o; --NO.sub.2; --CN; --N(R.sup.o).sub.2;
--NR.sup.oC(O)R.sup.o; --NR.sup.oC(S)R.sup.o;
--NR.sup.oC(O)N(R.sup.o).sub.2; --NR.sup.oC(S)N(R.sup.o).sub.2;
--NR.sup.oCO.sub.2R.sup.o; --NR.sup.oNR.sup.oC(O)R.sup.o;
--NR.sup.oNR.sup.oC(O)N(R.sup.o).sub.2;
--NR.sup.oNR.sup.oCO.sub.2R.sup.o; --C(O)C(O)R.sup.o;
--C(O)CH.sub.2C(O)R.sup.o; --CO.sub.2R.sup.o; --C(O)R.sup.o;
--C(S)R.sup.o; --C(O)N(R.sup.o).sub.2; --C(S)N(R.sup.o).sub.2;
--OC(O)N(R.sup.o).sub.2; --OC(O)R.sup.o; --C(O)N(OR.sup.o)R.sup.o;
--C(NOR.sup.o)R.sup.o; --S(O).sub.2R.sup.o; --S(O).sub.3R.sup.o;
--SO.sub.2N(R.sup.o).sub.2; --S(O)R.sup.o;
--NR.sup.oSO.sub.2N(R.sup.o).sub.2; --NR.sup.oSO.sub.2R.sup.o;
--N(OR.sup.o)R.sup.o; --C(.dbd.NH)--N(R.sup.o).sub.2; or
--(CH.sub.2).sub.0-2NHC(O)R.sup.o; wherein each independent
occurrence of R.sup.o is selected from hydrogen, optionally
substituted C.sub.1-6 aliphatic, an unsubstituted 5-6 membered
heteroaryl or heterocyclic ring, phenyl, --O(Ph), or
--CH.sub.2(Ph), or, two independent occurrences of R.sup.o, on the
same substituent or different substituents, taken together with the
atom(s) to which each R.sup.o group is bound, form a 5-8-membered
heterocyclyl, aryl, or heteroaryl ring or a 3-8-membered cycloalkyl
ring, wherein said heteroaryl or heterocyclyl ring has 1-3
heteroatoms independently selected from nitrogen, oxygen, or
sulfur. Optional substituents on the aliphatic group of R.sup.o are
selected from NH.sub.2, NH(C.sub.1-4aliphatic),
N(C.sub.1-4aliphatic).sub.2, halogen, C.sub.1-4aliphatic, OH,
O(C.sub.1-4aliphatic), NO.sub.2, CN, CO.sub.2H,
CO.sub.2(C.sub.1-4aliphatic), O(haloC.sub.1-4 aliphatic), or
haloC.sub.1-4aliphatic, wherein each of the foregoing
C.sub.1-4aliphatic groups of R.sup.o is unsubstituted.
[0026] In some embodiments, an aliphatic or heteroaliphatic group,
or a non-aromatic heterocyclic ring may contain one or more
substituents. Suitable substituents on the saturated carbon of an
aliphatic or heteroaliphatic group, or of a non-aromatic
heterocyclic ring are selected from those listed above for the
unsaturated carbon of an aryl or heteroaryl group and additionally
include the following: .dbd.O, .dbd.S, .dbd.NNHR*,
.dbd.NN(R*).sub.2, .dbd.NNHC(O)R*, .dbd.NNHCO.sub.2(alkyl),
.dbd.NNHSO.sub.2(alkyl), or .dbd.NR*, where each R* is
independently selected from hydrogen or an optionally substituted
C.sub.1-6 aliphatic. Optional substituents on the aliphatic group
of R* are selected from NH.sub.2, NH(C.sub.1-4 aliphatic),
N(C.sub.1-4 aliphatic).sub.2, halogen, C.sub.1-4 aliphatic, OH,
O(C.sub.1-4 aliphatic), NO.sub.2, CN, CO.sub.2H, CO.sub.2(C.sub.1-4
aliphatic), O(halo C.sub.1-4 aliphatic), or halo(C.sub.1-4
aliphatic), wherein each of the foregoing C.sub.1-4aliphatic groups
of R* is unsubstituted.
[0027] In some embodiments, optional substituents on the nitrogen
of a non-aromatic heterocyclic ring include --R.sup.+,
--N(R.sup.+).sub.2, --C(O)R.sup.+, --CO.sub.2R.sup.+,
--C(O)C(O)R.sup.+, --C(O)CH.sub.2C(O)R.sup.+, --SO.sub.2R.sup.+,
--SO.sub.2N(R.sup.+).sub.2, --C(.dbd.S)N(R.sup.+).sub.2,
--C(.dbd.NH)--N(R.sup.+).sub.2, or --NR.sup.+SO.sub.2R.sup.+;
wherein R.sup.+ is hydrogen, an optionally substituted C.sub.1-6
aliphatic, optionally substituted phenyl, optionally substituted
--O(Ph), optionally substituted --CH.sub.2(Ph), optionally
substituted --(CH.sub.2).sub.1-2(Ph); optionally substituted
--CH.dbd.CH(Ph); or an unsubstituted 5-6 membered heteroaryl or
heterocyclic ring having one to four heteroatoms independently
selected from oxygen, nitrogen, or sulfur, or, two independent
occurrences of R.sup.+, on the same substituent or different
substituents, taken together with the atom(s) to which each R.sup.+
group is bound, form a 5-8-membered heterocyclyl, aryl, or
heteroaryl ring or a 3-8-membered cycloalkyl ring, wherein said
heteroaryl or heterocyclyl ring has 1-3 heteroatoms independently
selected from nitrogen, oxygen, or sulfur. Optional substituents on
the aliphatic group or the phenyl ring of R.sup.+ are selected from
NH.sub.2, NH(C.sub.1-4 aliphatic), N(C.sub.1-4 aliphatic).sub.2,
halogen, C.sub.1-4 aliphatic, OH, O(C.sub.1-4 aliphatic), NO.sub.2,
CN, CO.sub.2H, CO.sub.2(C.sub.1-4 aliphatic), O(halo C.sub.1-4
aliphatic), or halo(C.sub.1-4 aliphatic), wherein each of the
foregoing C.sub.1-4aliphatic groups of R.sup.+ is
unsubstituted.
[0028] As detailed above, in some embodiments, two independent
occurrences of R.sup.o (or R.sup.+, or any other variable similarly
defined herein), may be taken together with the atom(s) to which
each variable is bound to form a 5-8-membered heterocyclyl, aryl,
or heteroaryl ring or a 3-8-membered cycloalkyl ring. Exemplary
rings that are formed when two independent occurrences of R.sup.o
(or R.sup.+, or any other variable similarly defined herein) are
taken together with the atom(s) to which each variable is bound
include, but are not limited to the following: a) two independent
occurrences of R.sup.o (or R.sup.+, or any other variable similarly
defined herein) that are bound to the same atom and are taken
together with that atom to form a ring, for example,
N(R.sup.o).sub.2, where both occurrences of R.sup.o are taken
together with the nitrogen atom to form a piperidin-1-yl,
piperazin-1-yl, or morpholin-4-yl group; and b) two independent
occurrences of R.sup.o (or R.sup.+, or any other variable similarly
defined herein) that are bound to different atoms and are taken
together with both of those atoms to form a ring, for example where
a phenyl group is substituted with two occurrences of OR.sup.o
##STR2## these two occurrences of R.sup.o are taken together with
the oxygen atoms to which they are bound to form a fused 6-membered
oxygen containing ring: ##STR3## It will be appreciated that a
variety of other rings can be formed when two independent
occurrences of R.sup.o (or R.sup.+, or any other variable similarly
defined herein) are taken together with the atom(s) to which each
variable is bound and that the examples detailed above are not
intended to be limiting.
[0029] In some embodiments, an alkyl or aliphatic chain can be
optionally interrupted with another atom or group. This means that
a methylene unit of the alkyl or aliphatic chain is optionally
replaced with said other atom or group. Examples of such atoms or
groups would include, but are not limited to, --NR--, --O--, --S--,
--CO.sub.2--, --OC(O)--, --C(O)CO--, --C(O)--, --C(O)NR--,
--C(.dbd.N--CN), --NRCO--, --NRC(O)O--, --SO.sub.2NR--,
--NRSO.sub.2--, --NRC(O)NR--, --OC(O)NR--, --NRSO.sub.2NR--,
--SO--, or --SO.sub.2--, wherein R is defined herein. Unless
otherwise specified, the optional replacements form a chemically
stable compound. Optional interruptions can occur both within the
chain and at either end of the chain; i.e. both at the point of
attachment and/or also at the terminal end. Two optional
replacements can also be adjacent to each other within a chain so
long as it results in a chemically stable compound. Unless
otherwise specified, if the replacement or interruption occurs at
the terminal end, the replacement atom is bound to an H on the
terminal end. For example, if --CH.sub.2CH.sub.2CH.sub.3 were
optionally interrupted with --O--, the resulting compound could be
--OCH.sub.2CH.sub.3, --CH.sub.2OCH.sub.3, or
--CH.sub.2CH.sub.2OH.
[0030] As described herein, a bond drawn from a substituent to the
center of one ring within a multiple-ring system (as shown below),
represents substitution of the substituent at any substitutable
position in any of the rings within the multiple ring system. For
example, Figure a represents possible substitution in any of the
positions shown in Figure b. ##STR4##
[0031] This also applies to multiple ring systems fused to optional
ring systems (which would be represented by dotted lines). For
example, in Figure c, X is an optional substituent both for ring A
and ring B. ##STR5##
[0032] If, however, two rings in a multiple ring system each have
different substituents drawn from the center of each ring, then,
unless otherwise specified, each substituent only represents
substitution on the ring to which it is attached. For example, in
Figure d, Y is an optionally substituent for ring A only, and X is
an optional substituent for ring B only. ##STR6##
[0033] Unless otherwise stated, structures depicted herein are also
meant to include all isomeric (e.g., enantiomeric, diastereomeric,
and geometric (or conformational)) forms of the structure; for
example, the R and S configurations for each asymmetric center, (Z)
and (E) double bond isomers, and (Z) and (E) conformational
isomers. Therefore, single stereochemical isomers as well as
enantiomeric, diastereomeric, and geometric (or conformational)
mixtures of the present compounds are within the scope of the
invention.
[0034] Unless otherwise stated, all tautomeric forms of the
compounds of the invention are within the scope of the invention.
Additionally, unless otherwise stated, structures depicted herein
are also meant to include compounds that differ only in the
presence of one or more isotopically enriched atoms. For example,
compounds having the present structures except for the replacement
of hydrogen by deuterium or tritium, or the replacement of a carbon
by a .sup.13C- or .sup.14C-enriched carbon are within the scope of
this invention. Such compounds are useful, for example, as
analytical tools or probes in biological assays.
[0035] As represented herein, the left-hand bond of Ring B is
attached to the bicyclic heteroaryl ring and the right-hand bond of
Ring B is attached to radical G.
[0036] Description of Compounds of the Invention:
[0037] The present invention relates to a compound of formula I:
##STR7## or a pharmaceutically acceptable salt thereof, wherein:
wherein ##STR8## [0038] Z.sub.1, Z.sub.2, Z.sub.3 and Z.sub.4 are
each independently selected from N or CR.sup.1, wherein at least
one of Z.sub.1, Z.sub.2, or Z.sub.4 is N; [0039] each R.sup.1 is
independently selected from H, halogen, --CN, --NO.sub.2, or
--V.sub.mR'; [0040] G is --NR.sup.2-- or --CO--; [0041] Q.sup.1 is
--CO--, --SO.sub.2--, --NR.sup.2, --NR.sup.2CO--, --CONR.sup.2--,
--SO.sub.2NR.sup.2--, or is a bond; [0042] R.sup.2 is --U.sub.nR'';
[0043] R.sup.3 is Q.sup.2-Ar.sup.1, or when G is --NR.sup.2,
R.sup.2 and Q.sup.1-R.sup.3, taken together with the nitrogen atom,
may form the cyclic group: ##STR9## where s is 1 or 2, Z is CH or
N; wherein each occurrence of Y is independently --CO--, --CS--,
--SO.sub.2--, --O--, --S--, --NR.sup.5--, or --C(R.sup.5).sub.2--,
and R.sup.5 is U.sub.nR'; [0044] X.sub.1 and X.sub.2 are each
independently selected from CR.sup.4 or N; [0045] each occurrence
of R.sup.4 is independently selected from halogen, CN, NO.sub.2, or
V.sub.mR; [0046] each occurrence of U or V is independently
selected from an optionally substituted C.sub.1-6 alkylidene chain,
wherein up to two methylene units of the chain are optionally and
independently replaced by --NR--, --S--, --O--, --CS--,
--CO.sub.2--, --OCO--, --CO--, --COCO--, --CONR--, --NRCO--,
--NRCO.sub.2--, --SO.sub.2NR--, --NRSO.sub.2--, --CONRNR--,
--NRCONR--, --OCONR--, --NRNR--, --NRSO.sub.2NR--, --SO--,
--SO.sub.2--, --PO--, --PO.sub.2--, or --POR--; [0047] m and n are
each independently 0 or 1; [0048] each occurrence of R is
independently selected from hydrogen or a C.sub.1-6 aliphatic
group, wherein said aliphatic group is optionally substituted with
up to five occurrences of J.sup.R; [0049] each occurrence of R' is
independently selected from hydrogen, a C.sub.1-6 aliphatic group,
a 3-8-membered saturated, partially unsaturated, or fully
unsaturated monocyclic ring having 0-3 heteroatoms independently
selected from nitrogen, oxygen, or sulfur, or an 8-12 membered
saturated, partially unsaturated, or fully unsaturated bicyclic
ring system having 0-5 heteroatoms independently selected from
nitrogen, oxygen, or sulfur, wherein said aliphatic group,
monocyclic ring or bicyclic ring is optionally substituted with up
to five occurrences of J.sup.R'; [0050] R'' is selected from
hydrogen, a C.sub.1-6 aliphatic group, a 3-8-membered saturated,
partially unsaturated, or fully unsaturated monocyclic ring having
0-3 heteroatoms independently selected from nitrogen, oxygen, or
sulfur, or an 8-12 membered saturated, partially unsaturated, or
fully unsaturated bicyclic ring system having 0-5 heteroatoms
independently selected from nitrogen, oxygen, or sulfur, wherein
said aliphatic group, monocyclic ring or bicyclic ring is
optionally substituted with up to five occurrences of J.sup.R'';
[0051] or two occurrences of R, R' and R'', in any combination
thereof, are taken together with the atom(s) to which they are
bound to form a 3-12 membered saturated, partially unsaturated, or
fully unsaturated monocyclic or bicyclic ring having 0-4
heteroatoms independently selected from nitrogen, oxygen, or
sulfur, wherein said monocyclic or bicyclic ring is optionally
substituted with J.sup.R; [0052] each occurrence of J.sup.R,
J.sup.R' and J.sup.R'' is independently selected from halogen, L,
-(L.sub.p)-R.sup.J, -(L.sub.p)-N(R.sup.J).sub.2,
-(L.sub.p)-SR.sup.J, -(L.sub.p)-OR.sup.J, -(L.sub.p)-(C.sub.3-10
cycloaliphatic), -(L.sub.p)-(C.sub.6-10 aryl), -(L.sub.p)-(5-10
membered heteroaryl), -(L.sub.p)-(5-10 membered heterocyclyl), oxo,
C.sub.1-4haloalkoxy, C.sub.1-4haloalkyl, -(L.sub.p)-NO.sub.2,
-(L.sub.p)-CN, -(L.sub.p)-OH, -(L.sub.p)-CF.sub.3,
--CO.sub.2R.sup.J, --CO.sub.2H, --COR.sup.J, --COH, --OC(O)R.sup.J
or --NC(O)R.sup.J; or any two J.sup.R, J.sup.R' or J.sup.R''
groups, on the same substituent or different substituents, together
with the atom(s) to which each J.sup.R, J.sup.R' or J.sup.R'' group
is bound, form a 5-7 membered saturated, unsaturated, or partially
saturated ring; [0053] R.sup.J is H or C.sub.1-6 aliphatic; or two
R.sup.J groups or an R.sup.J group and an R, R' or R'' group,
together with the atom to which they are attached, optionally form
a 3-6 membered cycloaliphatic or heterocyclyl, wherein said
aliphatic, cycloaliphatic or heterocyclyl is optionally substituted
with R*, --OR*, --SR*, --NO.sub.2, --CF.sub.3, --CN, --CO.sub.2R*,
--COR*, OCOR* or NHCOR*, wherein R* is H or an unsubstituted
C.sub.1-6 aliphatic; [0054] L is a C.sub.1-6 aliphatic wherein up
to three methylene units are replaced by --NH--, --NR.sup.L, --O--,
--S--, --CO.sub.2--, --OC(O)--, --C(O)CO--, --C(O)--, --C(O)NH--,
--C(O)NR.sup.6--, --C(.dbd.N--CN), --NHCO--, --NR.sup.LCO--,
--NHC(O)O--, --NR.sup.LC(O)O--, --SO.sub.2NH--,
--SO.sub.2NR.sup.L--, --NHSO.sub.2--, --NR.sup.LSO.sub.2--,
--NHC(O)NH--, --NR.sup.LC(O)NH--, --NHC(O)NR.sup.L--,
--NR.sup.LC(O)NR.sup.L, --OC(O)NH--, --OC(O)NR.sup.L--,
--NHSO.sub.2NH--, --NR.sup.LSO.sub.2NH--, --NHSO.sub.2NR.sup.L--,
--NR.sup.LSO.sub.2NR.sup.L, --SO-- or --SO.sub.2--; [0055] R.sup.L
is selected from C.sub.1-6 aliphatic, C.sub.3-10 cycloaliphatic,
C.sub.6-10 aryl, 5-10 membered heteroaryl or 5-10 membered
heterocyclyl; or two R.sup.L groups, on the same substituent or
different substituents, together with the atom(s) to which each
R.sup.L group is bound, form a 3-8 membered heterocyclyl; [0056]
each p is independently 0 or 1; [0057] Q.sup.2 and Q.sup.3 are each
independently selected from a bond or a C.sub.1-6 alkylidene chain,
wherein up to two methylene units of the chain are each optionally
and independently replaced by --NR'--, --S--, --O--, --CS--,
--CO.sub.2--, --OCO--, --CO--, --COCO--, --CONR'--, --NR'CO--,
--NR'CO.sub.2--, --SO.sub.2NR'--, --NR'SO.sub.2--, --CONR'NR'--,
--NR'CONR'--, --OCONR'--, --NR'NR'--, --NR'SO.sub.2NR'--, --SO--,
--SO.sub.2--, --PO--, --PO.sub.2--, or --POR'--; and wherein any
carbon atom in the one or more methylene units is optionally
substituted with one or two occurrences of R.sup.6, wherein each
occurrence of R.sup.6 is independently halogen, --CN, --NO.sub.2,
or --U.sub.nR', or two occurrences of R.sup.6, or R' and R.sup.6,
taken together with the atoms to which they are bound, form an
optionally substituted 3-6-membered cycloalkyl, heterocyclyl, aryl
or heteroaryl ring; and [0058] Ar.sup.1 and Ar.sup.2 are each
independently selected from a C.sub.1-6 aliphatic, a 3-8 membered
saturated, partially unsaturated, or fully unsaturated monocyclic
ring having 0-3 heteroatoms independently selected from nitrogen,
oxygen, or sulfur, or an 8-12 membered saturated, partially
unsaturated, or fully unsaturated bicyclic ring system having 0-5
heteroatoms independently selected from nitrogen, oxygen, or
sulfur; wherein Ar.sup.1 and Ar.sup.2 are each optionally
substituted with 0-5 independent occurrences of TR.sup.7; wherein T
is a bond or is a C.sub.1-C.sub.6 alkylidene chain wherein up to
two methylene units of T are optionally and independently replaced
by --NR--, --S--, --O--, --CS--, --CO.sub.2--, --OCO--, --CO--,
--COCO--, --CONR--, --NRCO--, --NRCO.sub.2--, --SO.sub.2NR--,
--NRSO.sub.2--, --CONRNR--, --NRCONR--, --OCONR--, --NRNR--,
--NRSO.sub.2NR--, --SO--, --SO.sub.2--, --PO--, --PO.sub.2--, or
--POR--; and each occurrence of R.sup.7 is independently selected
from --R', halogen, --NO.sub.2, --CN or .dbd.O.
[0059] In another embodiment, a compound of the invention has one
of formulae II-VII: ##STR10## ##STR11##
[0060] wherein R.sup.1Z1, R.sup.1Z2, R.sup.1Z3 and R.sup.1Z4 are
each independently selected from H, halogen, --CN, --NO.sub.2, or
--V.sub.mR'. In a further embodiment, the compound has one of
formulae II, III or VI.
[0061] In one embodiment, the compound is any one formuale I-VII
and ##STR12## In a further embodiment, the compound is either of
formulae II, III or VI. In another embodiment, X.sub.1 is CR.sup.4
and X.sub.2 is N or CR.sup.4. In a further embodiment, R.sup.4 is
H.
[0062] In one embodiment, the compound is any one of formuale I-VII
and R.sup.1 is independently selected from H, halogen or C.sub.1-3
aliphatic. In a further embodiment, the compound is either of
formulae II, III or VI and ##STR13## In yet a further embodiment,
R.sup.1 is independently selected from H or halogen.
[0063] In one embodiment of formulae II-VII, R.sup.1Z1, R.sup.1Z2,
R.sup.1Z3 and R.sup.1Z4, if present, are each independently
selected from H, halogen or C.sub.1-3 aliphatic. In a further
embodiment, R.sup.1Z1 and R.sup.1Z2, if present, are H. In yet
another embodiment, R.sup.1Z3 and R.sup.1Z4, if present, are H or
halogen. In a further embodiment, R.sup.1Z3 and R.sup.1Z4, if
present, are H or F and at least one of R.sup.1Z3 and R.sup.1Z4 is
H. In yet a further embodiment, R.sup.1Z3 and R.sup.1Z4, if
present, are H. In another embodiment, R.sup.1Z1, R.sup.1Z2,
R.sup.1Z3 and R.sup.1Z4, if present, are H.
[0064] In one embodiment, the compound is any one of formulae I-VII
and Q.sup.1 is --CO--, --SO.sub.2--, --NR.sup.2, --NR.sup.2CO--,
--CONR.sup.2--, --SO.sub.2NR.sup.2. In a further embodiment, the
compound is either of formulae II, III or VI and ##STR14## In a
further embodiment, G is --NR.sup.2 and Q.sup.1 is --CO--, or G is
--CO-- and Q.sup.1 is --NR.sup.2--. In yet a further embodiment,
R.sup.2 is H, --C.sub.1-4 aliphatic, -cyclopropyl,
(CH.sub.2).sub.1-3OH or ##STR15## In a still further embodiment,
R.sup.2 is H.
[0065] In another embodiment of the invention, the compound is any
one of formulae I-VII and R.sup.3 is Q.sup.2-Ar.sup.1. In a further
embodiment, the compound is either of formulae II, III or VI and
##STR16## In a further embodiment, Q.sup.2 is
--(CHR.sup.6).sub.q--, --(CHR.sup.6).sub.qO--,
--(CHR.sup.6).sub.qS--, --(CHR.sup.6).sub.qS(O).sub.2--,
--(CHR.sup.6).sub.qS(O)--, --(CHR.sup.6).sub.qNR--, or
--(CHR.sup.6).sub.qC(O)--, wherein q is 0, 1, 2, or 3, and each
R.sup.6 is R', --N(R)(R'), --(CH.sub.2).sub.1-4N(R)(R'),
--(CH.sub.2).sub.1-4C(CH.sub.3).sub.2N(R)(R'),
--(CH.sub.2).sub.1-4CH(CH.sub.3)N(R)(R'), --OR',
--(CH.sub.2).sub.1-4OR', --NR(CH.sub.2).sub.1-4N(R)(R'),
--NR(CH.sub.2).sub.1-4SO.sub.2R', --NR(CH.sub.2).sub.1-4COOR', or
--NR(CH.sub.2).sub.1-4COR', or two occurrences of R.sup.6, taken
together with the atoms to which they are bound, form an optionally
substituted 3-6-membered saturated, partially unsaturated, or fully
unsaturated ring. In yet a further embodiment, Q.sup.2 is
--(CHR.sup.6).sub.q--, q is 1 or 2, and R.sup.6 is R', --N(R)(R'),
--(CH.sub.2).sub.1-4N(R)(R'), --OR', --(CH.sub.2).sub.1-4OR' or
--NR(CH.sub.2).sub.1-4SO.sub.2R'. In a still further embodiment,
Q.sup.2 is --(CHR.sup.6).sub.q--, q is 1 or 2, and each R.sup.6 is
H.
[0066] In another embodiment, Ar.sup.1 is a C.sub.3-6 aliphatic, a
5-8 membered saturated, partially unsaturated, or fully unsaturated
monocyclic ring having 0-3 heteroatoms independently selected from
nitrogen, oxygen, or sulfur, or an 8-12 membered saturated,
partially unsaturated, or fully unsaturated bicyclic ring system
having 0-5 heteroatoms independently selected from nitrogen,
oxygen, or sulfur; wherein Ar.sup.1 is optionally substituted with
0-5 independent occurrences of TR.sup.7.
[0067] In a further embodiment, Ar.sup.1 is selected from one of
##STR17## ##STR18## ##STR19## ##STR20##
[0068] wherein t is 0, 1, 2, 3, 4, or 5, and wherein any Ar.sup.1
is bonded to Q through any substitutable nitrogen or carbon atom,
and wherein one or more hydrogen atoms on any substitutable
nitrogen or carbon atom is substituted with one or more independent
occurrences of TR.sup.7. In a further embodiment, Ar.sup.1 is
##STR21## In yet a further embodiment, Ar.sup.1 is ##STR22##
[0069] In some embodiments, t is 0, 1 or 2, and each TR.sup.7 is
independently selected from halogen, --CN, --R', --OR', --NRR',
--OSO.sub.2R', --NRSO.sub.2R', --NRSO.sub.2NRR', --SO.sub.2NRR',
--CONRR', --COR', --COOR', --NRCOR' or --SO.sub.2R'. In a further
embodiment, TR.sup.7 is selected from --F, --Cl, --CN, --NH.sub.2,
--CH.sub.3, --CH.sub.2CH.sub.3, --CH(CH.sub.3).sub.2, --OR.sup.x,
--OCF.sub.3, --NR.sup.xSO.sub.2R.sup.x,
--NR.sup.xSO.sub.2N(R.sup.x).sub.2, --COOC(CH.sub.3).sub.3,
--OSO.sub.2CH.sub.3, --OH, --SO.sub.2N(R.sup.x).sub.2,
--SO.sub.2N(R.sup.x).sub.2, --SO.sub.2R.sup.x, -pyrollidinone,
tetrahydrofuran or -D-(CH.sub.2).sub.p--Y, wherein R.sup.x is a H
or a C.sub.1-4 alkyl, D is --SO.sub.2--, --SO.sub.2NH--,
--NHSO.sub.2-- or --O--, p is 0-3, and Y is selected from:
##STR23## wherein R.sup.y is H or C.sub.1-3 alkyl, and wherein one
or more carbon atoms of Y is optionally substituted with
.dbd.O.
[0070] In one embodiment, t is 1 or 2 and one TR.sup.7 is
-D-(CH.sub.2).sub.p--Y, D is --O--, p is 2 or 3, Y is ##STR24## and
R.sup.y is H or CH.sub.3, and wherein one or more carbon atoms of Y
is optionally substituted with .dbd.O. In another embodiment, t is
1 or 2 and one TR.sup.7 is --SO.sub.2N(R.sup.x).sub.2,
--NR.sup.xSO.sub.2R.sup.x, --NHSO.sub.2R.sup.x, --OCF.sub.3, or
--OR.sup.x. In yet another embodiment, t is 1 or 2, and one or both
TR.sup.7 are F or Cl. In yet another embodiment, Ar.sup.1 is
##STR25## and t is 0 or 1.
[0071] Representative examples of compounds of formula I are set
forth below in Table 1 below. TABLE-US-00001 TABLE 1 Examples of
Compounds of Formula I: 1 ##STR26## 2 ##STR27## 3 ##STR28## 4
##STR29## 5 ##STR30## 6 ##STR31## 7 ##STR32## 8 ##STR33## 9
##STR34## 10 ##STR35## 11 ##STR36## 12 ##STR37## 13 ##STR38## 14
##STR39## 15 ##STR40## 16 ##STR41## 17 ##STR42## 18 ##STR43## 19
##STR44## 20 ##STR45##
[0072] General Synthetic Methodology
[0073] The compounds of this invention may be prepared in general
by methods known to those skilled in the art for analogous
compounds, as illustrated by the general schemes below, and the
preparative examples that follow.
[0074] Although certain exemplary embodiments are depicted and
described above and herein, it will be appreciated that compounds
of the invention can be prepared according to the methods described
generally above using appropriate starting materials by methods
generally available to one of ordinary skill in the art.
[0075] Uses, Formulation and Administration
[0076] As discussed above, the present invention provides compounds
that are inhibitors of protein kinases, and thus the present
compounds are useful for the treatment of diseases, disorders, and
conditions including, but not limited to a proliferative disorder,
a cardiac disorder, a neurodegenerative disorder, psychotic
disorders, an autoimmune disorder, a condition associated with
organ transplant, an inflammatory disorder, an immunologically
mediated disorder, a viral disease, or a bone disorder. In
preferred embodiments, the compounds are useful for the treatment
of hypertension, angina, angina pectoris, cerebrovascular
contraction, asthma, peripheral circulation disorder, premature
birth, cancer, erectile dysfunction, arteriosclerosis, spasm
(cerebral vasospasm and coronary vasospasm), retinopathy (e.g.,
glaucoma), inflammatory disorders, autoimmune disorders, AIDS,
osteoporosis, myocardial hypertrophy, ischemia/reperfusion-induced
injury, endothelial dysfunction, Alzheimer's disease, or benign
prostatic hyperplasia. In other embodiments, such conditions in
which ROCK is known to play a role include, without limitation,
hypertension, cerebral vasospasm, coronary vasospasm, bronchial
asthma, preterm labor, erectile dysfunction, glaucoma, vascular
smooth muscle cell proliferation, myocardial hypertrophy,
malignoma, ischemia/reperfusion-induced injury, endothelial
dysfunction, Crohn's Disease and colitis, neurite outgrowth,
Raynaud's Disease, angina, Alzheimer's disease, benign prostatic
hyperplasia, or atherosclerosis.
[0077] Accordingly, in another aspect of the present invention,
pharmaceutically acceptable compositions are provided, wherein
these compositions comprise any of the compounds as described
herein, and optionally comprise a pharmaceutically acceptable
carrier, adjuvant or vehicle. In certain embodiments, these
compositions optionally further comprise one or more additional
therapeutic agents.
[0078] It will also be appreciated that certain of the compounds of
present invention can exist in free form for treatment, or where
appropriate, as a pharmaceutically acceptable derivative thereof.
According to the present invention, a pharmaceutically acceptable
derivative includes, but is not limited to, pharmaceutically
acceptable prodrugs, salts, esters, salts of such esters, or any
other adduct or derivative which upon administration to a patient
in need is capable of providing, directly or indirectly, a compound
as otherwise described herein, or a metabolite or residue
thereof.
[0079] As used herein, the term "pharmaceutically acceptable salt"
refers to those salts which are, within the scope of sound medical
judgement, suitable for use in contact with the tissues of humans
and lower animals without undue toxicity, irritation, allergic
response and the like, and are commensurate with a reasonable
benefit/risk ratio. A "pharmaceutically acceptable salt" means any
non-toxic salt or salt of an ester of a compound of this invention
that, upon administration to a recipient, is capable of providing,
either directly or indirectly, a compound of this invention or an
inhibitorily active metabolite or residue thereof. As used herein,
the term "inhibitorily active metabolite or residue thereof" means
that a metabolite or residue thereof is also an inhibitor of a ROCK
kinase.
[0080] Pharmaceutically acceptable salts are well known in the art.
For example, S. M. Berge et al., describe pharmaceutically
acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66,
1-19, incorporated herein by reference. Pharmaceutically acceptable
salts of the compounds of this invention include those derived from
suitable inorganic and organic acids and bases. Examples of
pharmaceutically acceptable, nontoxic acid addition salts are salts
of an amino group formed with inorganic acids such as hydrochloric
acid, hydrobromic acid, phosphoric acid, sulfuric acid and
perchloric acid or with organic acids such as acetic acid, oxalic
acid, maleic acid, tartaric acid, citric acid, succinic acid or
malonic acid or by using other methods used in the art such as ion
exchange. Other pharmaceutically acceptable salts include adipate,
alginate, ascorbate, aspartate, benzenesulfonate, benzoate,
bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate,
cyclopentanepropionate, digluconate, dodecylsulfate,
ethanesulfonate, formate, fumarate, glucoheptonate,
glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate,
hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate,
laurate, lauryl sulfate, malate, maleate, malonate,
methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate,
oleate, oxalate, palmitate, pamoate, pectinate, persulfate,
3-phenylpropionate, phosphate, picrate, pivalate, propionate,
stearate, succinate, sulfate, tartrate, thiocyanate,
p-toluenesulfonate, undecanoate, valerate salts, and the like.
Salts derived from appropriate bases include alkali metal, alkaline
earth metal, ammonium and N.sup.+(C.sub.1-4alkyl).sub.4 salts. This
invention also envisions the quaternization of any basic
nitrogen-containing groups of the compounds disclosed herein. Water
or oil-soluble or dispersable products may be obtained by such
quaternization. Representative alkali or alkaline earth metal salts
include sodium, lithium, potassium, calcium, magnesium, and the
like. Further pharmaceutically acceptable salts include, when
appropriate, nontoxic ammonium, quaternary ammonium, and amine
cations formed using counterions such as halide, hydroxide,
carboxylate, sulfate, phosphate, nitrate, loweralkyl sulfonate and
aryl sulfonate.
[0081] As described above, the pharmaceutically acceptable
compositions of the present invention additionally comprise a
pharmaceutically acceptable carrier, adjuvant, or vehicle, which,
as used herein, includes any and all solvents, diluents, or other
liquid vehicle, dispersion or suspension aids, surface active
agents, isotonic agents, thickening or emulsifying agents,
preservatives, solid binders, lubricants and the like, as suited to
the particular dosage form desired. Remington's Pharmaceutical
Sciences, Sixteenth Edition, E. W. Martin (Mack Publishing Co.,
Easton, Pa., 1980) discloses various carriers used in formulating
pharmaceutically acceptable compositions and known techniques for
the preparation thereof. Except insofar as any conventional carrier
medium is incompatible with the compounds of the invention, such as
by producing any undesirable biological effect or otherwise
interacting in a deleterious manner with any other component(s) of
the pharmaceutically acceptable composition, its use is
contemplated to be within the scope of this invention. Some
examples of materials which can serve as pharmaceutically
acceptable carriers include, but are not limited to, ion
exchangers, alumina, aluminum stearate, lecithin, serum proteins,
such as human serum albumin, buffer substances such as phosphates,
glycine, sorbic acid, or potassium sorbate, partial glyceride
mixtures of saturated vegetable fatty acids, water, salts or
electrolytes, such as protamine sulfate, disodium hydrogen
phosphate, potassium hydrogen phosphate, sodium chloride, zinc
salts, colloidal silica, magnesium trisilicate, polyvinyl
pyrrolidone, polyacrylates, waxes,
polyethylene-polyoxypropylene-block polymers, wool fat, sugars such
as lactose, glucose and sucrose; starches such as corn starch and
potato starch; cellulose and its derivatives such as sodium
carboxymethyl cellulose, ethyl cellulose and cellulose acetate;
powdered tragacanth; malt; gelatin; talc; excipients such as cocoa
butter and suppository waxes; oils such as peanut oil, cottonseed
oil; safflower oil; sesame oil; olive oil; corn oil and soybean
oil; glycols; such a propylene glycol or polyethylene glycol;
esters such as ethyl oleate and ethyl laurate; agar; buffering
agents such as magnesium hydroxide and aluminum hydroxide; alginic
acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl
alcohol, and phosphate buffer solutions, as well as other non-toxic
compatible lubricants such as sodium lauryl sulfate and magnesium
stearate, as well as coloring agents, releasing agents, coating
agents, sweetening, flavoring and perfuming agents, preservatives
and antioxidants can also be present in the composition, according
to the judgment of the formulator.
[0082] In yet another aspect, a method for the treatment or
lessening the severity of a proliferative disorder, a cardiac
disorder, a neurodegenerative disorder, a psychotic disorder, an
autoimmune disorder, a condition associated with organ transplant,
an inflammatory disorder, an immunologically mediated disorder, a
viral disease, or a bone disorder is provided comprising
administering an effective amount of a compound, or a
pharmaceutically acceptable composition comprising a compound to a
subject in need thereof. In certain embodiments of the present
invention an "effective amount" of the compound or pharmaceutically
acceptable composition is that amount effective for treating or
lessening the severity of a proliferative disorder, a cardiac
disorder, a neurodegenerative disorder, a psychotic disorder, an
autoimmune disorder, a condition associated with organ transplant,
an inflammatory disorder, an immunologically mediated disorder, a
viral disease, or a bone disorder. The compounds and compositions,
according to the method of the present invention, may be
administered using any amount and any route of administration
effective for treating or lessening the severity of a proliferative
disorder, a cardiac disorder, a neurodegenerative disorder, an
autoimmune disorder, a condition associated with organ transplant,
an inflammatory disorder, an immunologically mediated disorder, a
viral disease, or a bone disorder. The exact amount required will
vary from subject to subject, depending on the species, age, and
general condition of the subject, the severity of the infection,
the particular agent, its mode of administration, and the like. The
compounds of the invention are preferably formulated in dosage unit
form for ease of administration and uniformity of dosage. The
expression "dosage unit form" as used herein refers to a physically
discrete unit of agent appropriate for the patient to be treated.
It will be understood, however, that the total daily usage of the
compounds and compositions of the present invention will be decided
by the attending physician within the scope of sound medical
judgment. The specific effective dose level for any particular
patient or organism will depend upon a variety of factors including
the disorder being treated and the severity of the disorder; the
activity of the specific compound employed; the specific
composition employed; the age, body weight, general health, sex and
diet of the patient; the time of administration, route of
administration, and rate of excretion of the specific compound
employed; the duration of the treatment; drugs used in combination
or coincidental with the specific compound employed, and like
factors well known in the medical arts. The term "patient", as used
herein, means an animal, preferably a mammal, and most preferably a
human.
[0083] The pharmaceutically acceptable compositions of this
invention can be administered to humans and other animals orally,
rectally, parenterally, intracisternally, intravaginally,
intraperitoneally, topically (as by powders, ointments, or drops),
bucally, as an oral or nasal spray, or the like, depending on the
severity of the infection being treated. In certain embodiments,
the compounds of the invention may be administered orally or
parenterally at dosage levels of about 0.01 mg/kg to about 50 mg/kg
and preferably from about 1 mg/kg to about 25 mg/kg, of subject
body weight per day, one or more times a day, to obtain the desired
therapeutic effect.
[0084] Liquid dosage forms for oral administration include, but are
not limited to, pharmaceutically acceptable emulsions,
microemulsions, solutions, suspensions, syrups and elixirs. In
addition to the active compounds, the liquid dosage forms may
contain inert diluents commonly used in the art such as, for
example, water or other solvents, solubilizing agents and
emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl
carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate,
propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in
particular, cottonseed, groundnut, corn, germ, olive, castor, and
sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene
glycols and fatty acid esters of sorbitan, and mixtures thereof.
Besides inert diluents, the oral compositions can also include
adjuvants such as wetting agents, emulsifying and suspending
agents, sweetening, flavoring, and perfuming agents.
[0085] Injectable preparations, for example, sterile injectable
aqueous or oleaginous suspensions may be formulated according to
the known art using suitable dispersing or wetting agents and
suspending agents. The sterile injectable preparation may also be a
sterile injectable solution, suspension or emulsion in a nontoxic
parenterally acceptable diluent or solvent, for example, as a
solution in 1,3-butanediol. Among the acceptable vehicles and
solvents that may be employed are water, Ringer's solution, U.S.P.
and isotonic sodium chloride solution. In addition, sterile, fixed
oils are conventionally employed as a solvent or suspending medium.
For this purpose any bland fixed oil can be employed including
synthetic mono- or diglycerides. In addition, fatty acids such as
oleic acid are used in the preparation of injectables.
[0086] The injectable formulations can be sterilized, for example,
by filtration through a bacterial-retaining filter, or by
incorporating sterilizing agents in the form of sterile solid
compositions which can be dissolved or dispersed in sterile water
or other sterile injectable medium prior to use.
[0087] In order to prolong the effect of a compound of the present
invention, it is often desirable to slow the absorption of the
compound from subcutaneous or intramuscular injection. This may be
accomplished by the use of a liquid suspension of crystalline or
amorphous material with poor water solubility. The rate of
absorption of the compound then depends upon its rate of
dissolution that, in turn, may depend upon crystal size and
crystalline form. Alternatively, delayed absorption of a
parenterally administered compound form is accomplished by
dissolving or suspending the compound in an oil vehicle. Injectable
depot forms are made by forming microencapsule matrices of the
compound in biodegradable polymers such as
polylactide-polyglycolide. Depending upon the ratio of compound to
polymer and the nature of the particular polymer employed, the rate
of compound release can be controlled. Examples of other
biodegradable polymers include poly(orthoesters) and
poly(anhydrides). Depot injectable formulations are also prepared
by entrapping the compound in liposomes or microemulsions that are
compatible with body tissues.
[0088] Compositions for rectal or vaginal administration are
preferably suppositories which can be prepared by mixing the
compounds of this invention with suitable non-irritating excipients
or carriers such as cocoa butter, polyethylene glycol or a
suppository wax which are solid at ambient temperature but liquid
at body temperature and therefore melt in the rectum or vaginal
cavity and release the active compound.
[0089] Solid dosage forms for oral administration include capsules,
tablets, pills, powders, and granules. In such solid dosage forms,
the active compound is mixed with at least one inert,
pharmaceutically acceptable excipient or carrier such as sodium
citrate or dicalcium phosphate and/or a) fillers or extenders such
as starches, lactose, sucrose, glucose, mannitol, and silicic acid,
b) binders such as, for example, carboxymethylcellulose, alginates,
gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants
such as glycerol, d) disintegrating agents such as agar--agar,
calcium carbonate, potato or tapioca starch, alginic acid, certain
silicates, and sodium carbonate, e) solution retarding agents such
as paraffin, f) absorption accelerators such as quaternary ammonium
compounds, g) wetting agents such as, for example, cetyl alcohol
and glycerol monostearate, h) absorbents such as kaolin and
bentonite clay, and i) lubricants such as talc, calcium stearate,
magnesium stearate, solid polyethylene glycols, sodium lauryl
sulfate, and mixtures thereof. In the case of capsules, tablets and
pills, the dosage form may also comprise buffering agents.
[0090] Solid compositions of a similar type may also be employed as
fillers in soft and hard-filled gelatin capsules using such
excipients as lactose or milk sugar as well as high molecular
weight polyethylene glycols and the like. The solid dosage forms of
tablets, dragees, capsules, pills, and granules can be prepared
with coatings and shells such as enteric coatings and other
coatings well known in the pharmaceutical formulating art. They may
optionally contain opacifying agents and can also be of a
composition that they release the active ingredient(s) only, or
preferentially, in a certain part of the intestinal tract,
optionally, in a delayed manner. Examples of embedding compositions
that can be used include polymeric substances and waxes. Solid
compositions of a similar type may also be employed as fillers in
soft and hard-filled gelatin capsules using such excipients as
lactose or milk sugar as well as high molecular weight polethylene
glycols and the like.
[0091] The active compounds can also be in micro-encapsulated form
with one or more excipients as noted above. The solid dosage forms
of tablets, dragees, capsules, pills, and granules can be prepared
with coatings and shells such as enteric coatings, release
controlling coatings and other coatings well known in the
pharmaceutical formulating art. In such solid dosage forms the
active compound may be admixed with at least one inert diluent such
as sucrose, lactose or starch. Such dosage forms may also comprise,
as is normal practice, additional substances other than inert
diluents, e.g., tableting lubricants and other tableting aids such
a magnesium stearate and microcrystalline cellulose. In the case of
capsules, tablets and pills, the dosage forms may also comprise
buffering agents. They may optionally contain opacifying agents and
can also be of a composition that they release the active
ingredient(s) only, or preferentially, in a certain part of the
intestinal tract, optionally, in a delayed manner. Examples of
embedding compositions that can be used include polymeric
substances and waxes.
[0092] Dosage forms for topical or transdermal administration of a
compound of this invention include ointments, pastes, creams,
lotions, gels, powders, solutions, sprays, inhalants or patches.
The active component is admixed under sterile conditions with a
pharmaceutically acceptable carrier and any needed preservatives or
buffers as may be required. Ophthalmic formulation, ear drops, and
eye drops are also contemplated as being within the scope of this
invention. Additionally, the present invention contemplates the use
of transdermal patches, which have the added advantage of providing
controlled delivery of a compound to the body. Such dosage forms
can be made by dissolving or dispensing the compound in the proper
medium. Absorption enhancers can also be used to increase the flux
of the compound across the skin. The rate can be controlled by
either providing a rate controlling membrane or by dispersing the
compound in a polymer matrix or gel.
[0093] As described generally above, the compounds of the invention
are useful as inhibitors of protein kinases. In one embodiment, the
compounds and compositions of the invention are inhibitors of ROCK,
and thus, without wishing to be bound by any particular theory, the
compounds and compositions are particularly useful for treating or
lessening the severity of a disease, condition, or disorder where
activation of ROCK is implicated in the disease, condition, or
disorder. When activation of ROCK is implicated in a particular
disease, condition, or disorder, the disease, condition, or
disorder may also be referred to as "ROCK-mediated disease" or
disease symptom. Accordingly, in another aspect, the present
invention provides a method for treating or lessening the severity
of a disease, condition, or disorder where activation of ROCK is
implicated in the disease state.
[0094] The activity of a compound utilized in this invention as an
inhibitor of ROCK, may be assayed in vitro, in vivo or in a cell
line. In vitro assays include assays that determine inhibition of
either the phosphorylation activity or ATPase activity of activated
ROCK. Alternate in vitro assays quantitate the ability of the
inhibitor to bind to ROCK. Inhibitor binding may be measured by
radiolabelling the inhibitor prior to binding, isolating the
inhibitor/ROCK complex and determining the amount of radiolabel
bound. Alternatively, inhibitor binding may be determined by
running a competition experiment where new inhibitors are incubated
with ROCK bound to known radioligands.
[0095] The term "measurably inhibit", as used herein means a
measurable change in ROCK activity between a sample comprising said
composition and ROCK and an equivalent sample comprising ROCK in
the absence of said composition.
[0096] The term "ROCK-mediated condition" or "disease", as used
herein, means any disease or other deleterious condition in which
ROCK is known to play a role. The term "ROCK-mediated condition" or
"disease" also means those diseases or conditions that are
alleviated by treatment with a ROCK inhibitor. Such conditions
include, without limitation, hypertension, angina, angina pectoris,
cerebrovascular contraction, asthma, peripheral circulation
disorder, premature birth, cancer, erectile dysfunction,
arteriosclerosis, spasm (cerebral vasospasm and coronary
vasospasm), retinopathy (e.g., glaucoma), inflammatory disorders,
autoimmune disorders, AIDS, osteoporosis, myocardial hypertrophy,
ischemia/reperfusion-induced injury, endothelial dysfunction,
Alzheimer's disease, or benign prostatic hyperplasia. In other
embodiments, such conditions in which ROCK is known to play a role
include, without limitation, hypertension, cerebral vasospasm,
coronary vasospasm, bronchial asthma, preterm labor, erectile
dysfunction, glaucoma, vascular smooth muscle cell proliferation,
myocardial hypertrophy, malignoma, ischemia/reperfusion-induced
injury, endothelial dysfunction, Crohn's Disease and colitis,
neurite outgrowth, Raynaud's Disease, angina, Alzheimer's disease,
benign prostatic hyperplasia, or atherosclerosis.
[0097] It will also be appreciated that the compounds and
pharmaceutically acceptable compositions of the present invention
can be employed in combination therapies, that is, the compounds
and pharmaceutically acceptable compositions can be administered
concurrently with, prior to, or subsequent to, one or more other
desired therapeutics or medical procedures. The particular
combination of therapies (therapeutics or procedures) to employ in
a combination regimen will take into account compatibility of the
desired therapeutics and/or procedures and the desired therapeutic
effect to be achieved. It will also be appreciated that the
therapies employed may achieve a desired effect for the same
disorder (for example, an inventive compound may be administered
concurrently with another agent used to treat the same disorder),
or they may achieve different effects (e.g., control of any adverse
effects). As used herein, additional therapeutic agents that are
normally administered to treat or prevent a particular disease, or
condition, are known as "appropriate for the disease, or condition,
being treated".
[0098] For example, chemotherapeutic agents or other
anti-proliferative agents may be combined with the compounds of
this invention to treat proliferative diseases and cancer. Examples
of known chemotherapeutic agents include, but are not limited to,
For example, other therapies or anticancer agents that may be used
in combination with the inventive anticancer agents of the present
invention include surgery, radiotherapy (in but a few examples,
gamma.-radiation, neutron beam radiotherapy, electron beam
radiotherapy, proton therapy, brachytherapy, and systemic
radioactive isotopes, to name a few), endocrine therapy, biologic
response modifiers (interferons, interleukins, and tumor necrosis
factor (TNF) to name a few), hyperthermia and cryotherapy, agents
to attenuate any adverse effects (e.g., antiemetics), and other
approved chemotherapeutic drugs, including, but not limited to,
alkylating drugs (mechlorethamine, chlorambucil, Cyclophosphamide,
Melphalan, Ifosfamide), antimetabolites (Methotrexate), purine
antagonists and pyrimidine antagonists (6-Mercaptopurine,
5-Fluorouracil, Cytarabile, Gemcitabine), spindle poisons
(Vinblastine, Vincristine, Vinorelbine, Paclitaxel),
podophyllotoxins (Etoposide, Irinotecan, Topotecan), antibiotics
(Doxorubicin, Bleomycin, Mitomycin), nitrosoureas (Carmustine,
Lomustine), inorganic ions (Cisplatin, Carboplatin), enzymes
(Asparaginase), and hormones (Tamoxifen, Leuprolide, Flutamide, and
Megestrol), Gleevec.TM., adriamycin, dexamethasone, and
cyclophosphamide. For a more comprehensive discussion of updated
cancer therapies see, http://www.nci.nih.gov/, a list of the FDA
approved oncology drugs at
http://www.fda.gov/cder/cancer/druglistframe.htm, and The Merck
Manual, Seventeenth Ed. 1999, the entire contents of which are
hereby incorporated by reference.
[0099] Other examples of agents the inhibitors of this invention
may also be combined with include, without limitation: treatments
for Alzheimer's Disease such as Aricept.RTM. and Excelon.RTM.;
treatments for Parkinson's Disease such as L-DOPA/carbidopa,
entacapone, ropinrole, pramipexole, bromocriptine, pergolide,
trihexephendyl, and amantadine; agents for treating Multiple
Sclerosis (MS) such as beta interferon (e.g., Avonex.RTM. and
Rebif.RTM.), Copaxone.RTM., and mitoxantrone; treatments for asthma
such as albuterol and Singulair.RTM.; agents for treating
schizophrenia such as zyprexa, risperdal, seroquel, and
haloperidol; anti-inflammatory agents such as corticosteroids, TNF
blockers, IL-1 RA, azathioprine, cyclophosphamide, and
sulfasalazine; immunomodulatory and immunosuppressive agents such
as cyclosporin, tacrolimus, rapamycin, mycophenolate mofetil,
interferons, corticosteroids, cyclophosphamide, azathioprine, and
sulfasalazine; neurotrophic factors such as acetylcholinesterase
inhibitors, MAO inhibitors, interferons, anti-convulsants, ion
channel blockers, riluzole, and anti-Parkinsonian agents; agents
for treating cardiovascular disease such as beta-blockers, ACE
inhibitors, diuretics, nitrates, calcium channel blockers, and
statins; agents for treating liver disease such as corticosteroids,
cholestyramine, interferons, and anti-viral agents; agents for
treating blood disorders such as corticosteroids, anti-leukemic
agents, and growth factors; and agents for treating
immunodeficiency disorders such as gamma globulin.
[0100] The amount of additional therapeutic agent present in the
compositions of this invention will be no more than the amount that
would normally be administered in a composition comprising that
therapeutic agent as the only active agent. Preferably the amount
of additional therapeutic agent in the presently disclosed
compositions will range from about 50% to 100% of the amount
normally present in a composition comprising that agent as the only
therapeutically active agent.
[0101] The compounds of this invention or pharmaceutically
acceptable compositions thereof may also be incorporated into
compositions for coating implantable medical devices, such as
prostheses, artificial valves, vascular grafts, stents and
catheters. Accordingly, the present invention, includes a
composition for coating an implantable device comprising a compound
of the present invention as described generally above, and in
classes and subclasses herein, and a carrier suitable for coating
said implantable device. In still another aspect, the present
invention includes an implantable device coated with a composition
comprising a compound of the present invention as described
generally above and a carrier suitable for coating said implantable
device.
[0102] Vascular stents, for example, have been used to overcome
restenosis. However, patients using stents or other implantable
devices risk clot formation or platelet activation. These unwanted
effects may be prevented or mitigated by pre-coating the device
with a pharmaceutically acceptable composition comprising a kinase
inhibitor. Suitable coatings and the general preparation of coated
implantable devices are described in U.S. Pat. Nos. 6,099,562;
5,886,026; and 5,304,121. The coatings are typically biocompatible
polymeric materials such as a hydrogel polymer,
polymethyldisiloxane, polycaprolactone, polyethylene glycol,
polylactic acid, ethylene vinyl acetate, and mixtures thereof. The
coatings may optionally be further covered by a suitable topcoat of
fluorosilicone, polysaccarides, polyethylene glycol, phospholipids
or combinations thereof to impart controlled release
characteristics in the composition.
[0103] Another aspect of the invention relates to inhibiting ROCK
activity in a biological sample or a patient, which method
comprises administering to the patient, or contacting said
biological sample with a compound of formula I or a composition
comprising said compound. The term "biological sample", as used
herein, includes, without limitation, cell cultures or extracts
thereof; biopsied material obtained from a mammal or extracts
thereof; and blood, saliva, urine, feces, semen, tears, or other
body fluids or extracts thereof. Inhibition of ROCK kinase activity
in a biological sample is useful for a variety of purposes that are
known to one of skill in the art. Examples of such purposes
include, but are not limited to, blood transfusion,
organ-transplantation, biological specimen storage, and biological
assays.
[0104] All references provided in the Examples are herein
incorporated by reference. As used herein, all abbreviations,
symbols and conventions are consistent with those used in the
contemporary scientific literature. See, e.g., Janet S. Dodd, ed.,
The ACS Style Guide: A Manual for Authors and Editors, 2nd Ed.,
Washington, D.C.: American Chemical Society, 1997, herein
incorporated in its entirety by reference.
EXAMPLES
Example 1
Preparation of Compound 9
[0105] ##STR46## 4-Bromothiophene-2-carboxylic acid (A)
[0106] A solution of sodium chlorite (47.5 g, 525 mmol) and sodium
hydrogen phosphate (63.0 g, 525 mmol) in water (500 mL) was added
slowly to a stirred solution of 4-bromo-2-thiophenecarboxaldehyde
(50 g, 261 mmol) in tertiary butanol (600 mL) and 2-methylbutene (8
mL) in 30 min at 0.degree. C. The cooling bath was removed and the
resulting solution was stirred at room temperature for 16 h. The
mixture was transferred to a separator funnel and the aqueous layer
was separated. Organic layer (tertiary butanol) was concentrated
under reduced pressure to give white residue, which was added to
the aqueous aliquot. The total aqueous layer was acidified with 6N
HCl (100 mL). The precipitated product was extracted with ethyl
acetate (3.times.250 mL). The organic layer was dried
(Na.sub.2SO.sub.4) and concentrated under reduced pressure to
afford the title 4-bromothiophene-2-carboxylic acid (A) as a white
solid (54 g, 100%). FIA MS 207 (M+1).
tert-Butyl-4-bromothiophen-2-yl-carbamate (B)
[0107] A mixture of 4-bromothiophene-2-carboxylic acid (A) (53 g,
255 mmol), diphenylphosphoryl azide (Aldrich, 70 mL, 323 mmol),
triethylamine (45 mmol) in tertiary butanol (675 mL) was heated at
100.degree. C. for 5 h and then cooled to room temperature. The
solvent was evaporated to give brown gum, which was dissolved in
EtOAc (500 mL). The organic solution was washed with saturated
NaHCO.sub.3 (500 mL) and water (500 mL) respectively, then dried
over Na.sub.2SO.sub.4 and concentrated. The crude product was
dissolved in CH.sub.2Cl.sub.2 (75 mL) and purified by flash column
chromatography on silica gel (10%-15% EtOAc/hexanes) to afford
title compound tert-butyl-4-bromothiophen-2-yl-carbamate (B) (47 g,
66%) as a white solid. FIA MS 278 (M+1).
5-BOC.sup.taminothiophen-3-yl-3-boronic acid (C)
[0108] 2.5M nBuLi in hexane (7.2 mL, 18 mmol) was added dropwise to
a stirred solution of tert-butyl-4-bromothiophen-2-yl-carbamate (B)
(1 g, 3.6 mmol) in THF (3 mL) and toluene (11 mL) at -78.degree. C.
under nitrogen and the solution was stirred 1 h at -78.degree.
C.
[0109] Triisopropylborate (2 mL, 9 mmol) was added and the
resulting thick brown solution was stirred at -78.degree. C. for 30
min and room temperature for 30 min. 2N HCl (15 ml) and ethyl
acetate (50 mL) were added and the solution was stirred 25 min. The
organic layer was separated and aqueous layer was extracted with
EtOAc (2.times.25 mL). The combined organic layers were dried
(Na.sub.2SO.sub.4) and concentrated to give title compound
5-aminothiophen-3-yl-3-boronic acid (C) as an oil (0.8 g, 91%). The
product was used for next step without purification. FIA MS 242
(M-1).
5-Aminothiophen-3-yl-3-boronic acid (D)
[0110] The crude 5-aminothiophen-3-yl-3-boronic acid (C) (0.8 g)
was dissolved in 4N HCl (10 mL) and methanol (2 mL) and the
solution was stirred at room temperature for 2 h. The solvent was
evaporated under reduced pressure to give title compound
5-BOC.sup.taminothiophen-3-yl-3-boronic acid (D) (0.58 g) as a
brown gum. The product was used for next step without purification.
FIA MS 143 (M+1).
5-(2-(3-Methsulfonamidophenyl)acetamido)thiophen-3-yl-3-boronic
acid (E)
[0111] A mixture of 5-aminothiophen-3-yl-3-boronic acid (D) (0.48
g, 3.35 mmol), 3-methylsulfonamidephenylacetic acid (0.77 g, 3.35
mmol), BtSO.sub.2CH.sub.3 (0.66 g, 3.35 mmol), Et.sub.3N (3 mL) in
THF (25 mL) and DMF (2 mL) was heated at 85.degree. C. for 16 h and
then cooled to room temperature. The solvent was evaporated and the
crude material was dissolved in EtOAc (50 mL). The organic solution
was washed with saturated NaHCO.sub.3 (20 mL), brine (20 mL), dried
(Na.sub.2SO.sub.4) and concentrated. The crude material was
purified by chromatography. The impurities were first eluted with
40-70% EtOAc/hexanes and the product was eluted with 5-30%
MeOH/CH.sub.2Cl.sub.2. Yield 0.54 g, 46%, FIA MS 355 (M+1).
5-(2-(5-Methsulfonamido-2-fluorophenyl)acetamido)thiophen-3-yl-3-boronic
acid (F)
[0112] The boronic acid F was synthesized and purified in a manner
similar to E using a mixture of D (0.48 g, 3.35 mmol),
3-methylsulfonamide-6-flurophenylacetic acid (0.82 g, 3.35 mmol),
BtSO.sub.2CH.sub.3 (0.66 g, 3.35 mmol), Et.sub.3N (3 mL). Yield
0.43 g, 34%, FIA MS 373 (M+1).
5-(2-(3-N,N-dimethylaminosulfonylbenzene)acetamido)thiophen-3-yl-3-boron-
ic acid (G)
[0113] The boronic acid F was synthesized and purified in a manner
similar to E using a mixture of D (0.48 g, 3.35 mmol),
3-N,N-dimethylsulfone phenylacetic acid (0.81 g, 3.35 mmol),
BtSO.sub.2CH.sub.3 (0.66 g, 3.35 mmol), Et.sub.3N (3 mL). Yield
0.64 g, 52%, FIA MS 369 (M+1).
5-(2-(3-(3-(tert-butoxycarbonyl-piperidin-4-yl)propoxy)phenyl)acetamido)-
thiophene-3-yl-3-boronic acid (H)
[0114] The boronic acid H was synthesized and purified in a manner
similar to E using a mixture of D (0.48 g, 3.35 mmol),
2-(3-(3-(BOC.sup.tpiperidin-4-yl)propoxy)phenylacetic acid (1.26 g,
3.35 mmol), BtSO.sub.2CH.sub.3 (0.66 g, 3.35 mmol), Et.sub.3N (3
mL). Yield 0.57 g, 43.6%, FIA MS 503 (M+1). ##STR47##
N-(4-(1H-pyrazolo[3,4-b]pyrazin-3-yl)thiophen-2-yl)-2-(5-methsulfonamido--
2-fluorophenyl)acetamide (Compound 9)
[0115] A mixture of J (0.050 g, 0.134 mmol), M (see Monatshefte fur
Chemie 113:731, 1992; 0.026 g, 0.134 mmol), Pd.sub.2(dba).sub.3
(0.017 g, 0.02 mmol), P(Bu.sup.t).sub.3 (0.01 g, 0.04 mmol),
KF.H.sub.2O (0.1 g, 1.3 mmol) in 1,4-dioxane (2 mL) and water (0.5
mL) was heated at 185.degree. C. in a microwave for 25 min. Solid
was filtered and washed with EtOAc (1 mL). The filtrate was
concentrated and the crude product was purified by preparative HPLC
(5-75% CH.sub.3CN/water, 15 min) to give title compound 9 (0.005 g)
as an oil.
Example 2
Preparation of Compounds 6, 7, 8 and 10
[0116] ##STR48## 1 Fluropyridine-3-carbaldehyde (N)
[0117] Under nitrogen, 2.5M nBuLi in hexane (25.6 mL, 64.37 mmol)
was added dropwise to a stirred solution of isopropylamine (9 mL,
64 mmol) in THF (50 mL) and at -78.degree. C. for 10 min. and the
solution was stirred 30 min at -78.degree. C. 2-Fluropyridine (4.4
mL, 51.2 mmol) was added and the solution was stirred at
-78.degree. C. for 2 h. A cold solution of
N-methyl-N-(pyridin-2-yl)formamide (7 g, 51.2 mmol) in THF (30 mL)
were added in 15 min. The resulting solution was warmed to room
temperature and stirred for 1 h. 2N HCl (15 mL) and ethyl acetate
(50 mL) ware added and the solution was stirred 25 min. The organic
layer was separated and aqueous layer was extracted with EtOAc
(2.times.25 mL). The combined organic layers were dried
(Na.sub.2SO.sub.4) and concentrated. The crude product was purified
by chromatography (10-60% EtOAc/hexanes) to give title compound N
(1.3 g, 20%) as a yellow oil. .sup.1H NMR (DMSO-d6, 500 MHz)
.delta. 10.16(s, 1H), 8.53(dd, 1H), 8.38(m, 1H), 7.58(m, 1H).
(E)-N'-((2-Fluoropyridin-3-yl)methylene)acetohydrazide (O)
[0118] A solution of N (1.3 g, 10.4 mmol), acylhydrazine (1.5 g,
20.8 mmol) in ethanol (10 mL) was stirred at room temperature for
18 h. The precipitate was filtered and washed with ethanol
(2.times.10 mL) and dried to give title compound O (0.82 g, 43%) as
a white solid FIA MS 180 (M-1).
1H-Pyrazolo[3,4-b]pyridine (P)
[0119] A suspension of O (0.82 g, 4.5 mmol) in hydrazine
monohydrate (5 mL) was heated at 90.degree. C. for 15 min and
poured into ice/water and extracted with ethyl acetate (3.times.25
mL). The organic layers were dried (Na.sub.2SO.sub.4) and
concentrated to give title compound P (0.41 g, 77%) as a white
solid. .sup.1H NMR (DMSO-d.sub.6, 500 MHz) .delta. 13.60(brs, 1H),
8.55(d, 1H), 8.51(dd, 1H), 8.14(s, 1H), 7.18(dd, 1H); FIA MS 120
(M+1).
3-Bromo-1H-pyrazolo[3,4-b]pyridine (Q)
[0120] Bromine (0.3 mL, 5.16 mmol) was added via a micro syringe to
a stirred solution of P (0.41 g, 3.45 mmol) in chloroform (10 mL).
The resulting suspension was stirred at room temperature for 2 h.
The solvent was evaporated and ethyl acetate (50 mL) was added. The
organic layer was washed with saturated K.sub.2CO.sub.3 and the
organic layer was concentrated without drying at 70.degree. C. to
produce title compound Q (0.63 g, 92%). .sup.1H NMR (DMSO-d.sub.6,
500 MHz) .delta. 8.56(t, 1H), 8.05(d, 1H), 7.24(m, 1H); FIA MS
198(M-1).
3-Bromo-1-tosyl-1H-pyrazolo[3,4-b]pyridine (R)
[0121] Sodium hydride (0.05 g, 2 mmol) was added to a stirred
solution of Q (0.1 g, 0.5 mmol) in THF (5 mL). The solution was
stirred at room temperature for 30 min and P-TsCl (0.1 g, 0.5 mmol)
was added. The resulting suspension was heated at 60.degree. C. for
45 min. and poured into water (25 mL). The solution was extracted
with ethyl acetate (2.times.25 mL), dried and concentrated to give
title compound R (0.14, 80%). The product was unstable and
therefore used for next step immediately. FIA-MS 353(M-1).
2-(2-Fluoro-5-methanesulfonylamino-phenyl)-N-[4-(1H-pyrazolo[3,4-b]pyrid-
in-3-yl)-thiophen-2-yl]-acetamide (Compound 6).
[0122] A mixture of R (0.050 g, 0.142 mmol), F (0.053 g, 0.142
mmol), Pd.sub.2(dba).sub.3 (0.017 g, 0.02 mmol), P(Bu.sup.t).sub.3
(0.01 g, 0.04 mmol), KF.H.sub.2O (0.1 g, 1.3 mmol) in 1,4-dioxane
(2 mL) and water (0.5 mL) was heated at 185.degree. C. in a
microwave for 25 min. Solid was filtered and washed with EtOAC (1
mL). The filtrate was concentrated and the crude product was
purified by preparative HPLC (5-75% CH.sub.3CN/water, 15 min) to
give title compound 6 (0.027 g) as an oil.
2-(3-Methanesulfonylamino-phenyl)-N-[4-(1H-pyrazolo[3,4-b]pyridin-3-yl)--
thiophen-2-yl]-acetamide (Compound 8).
[0123] The title compound 8 was synthesized and purified in a
manner similar to 6 using a mixture of R (0.050 g, 0.142 mmol), E
(0.050 g, 0.142 mmol), Pd.sub.2(dba).sub.3 (0.017 g, 0.02 mmol),
P(Bu.sup.t).sub.3 (0.01 g, 0.04 mmol), KF.H.sub.2O (0.1 g, 1.3
mmol) in 1,4-dioxane (2 mL) and water (0.5 mL). Yield 0.03 g.
2-(3-Dimethylsulfamoyl-phenyl)-N-[4-(1H-pyrazolo[3,4-b]pyridin-3-yl)-thi-
ophen-2-yl]-acetamide (Compound 10)
[0124] The title compound 10 was synthesized and purified in a
manner similar to 6 using a mixture of R (0.050 g, 0.142 mmol), G
(0.052 g, 0.142 mmol), Pd.sub.2(dba).sub.3 (0.017 g, 0.02 mmol),
P(Bu.sup.t).sub.3 (0.01 g, 0.04 mmol), KF.H.sub.2O (0.1 g, 1.3
mmol) in 1,4-dioxane (2 mL) and water (0.5 mL). Yield 0.026 g.
2-(3-(3-Piperidin-4-yl-propoxy)-phenyl)-N-[4-(1H-pyrazolo[3,4-b]pyridin--
3-yl)-thiophen-2-yl]-acetamide (Compound 7)
[0125] A mixture of R (0.050 g, 0.142 mmol), H (0.064 g, 0.142
mmol), Pd.sub.2(dba).sub.3 (0.017 g, 0.02 mmol), P(Bu.sup.t).sub.3
(0.01 g, 0.04 mmol), KF.H.sub.2O (0.1 g, 1.3 mmol) in 1,4-dioxane
(2 mL) and water (0.5 mL) was heated at 185.degree. C. in a
microwave for 25 min. Solid was filtered and washed with EtOAC (1
mL). The filtrate was concentrated. The crude product was dissolved
in CH.sub.2Cl.sub.2 (1 mL) and TFA (1 mL). The solution was stirred
at room temperature for 1 h. and concentrated. The crude product
was purified by preparative HPLC (5-75% CH.sub.3CN/water, 15 min)
to give title compound 7 (0.032 g) as an oil.
Example 3
Preparation of Compounds 19 and 20
[0126] ##STR49##
5-Bromo-3-trimethylsilanylethynyl-pyrazin-2-ylamine (U)
[0127] Trimethylsilylacetylene (3.4 mL) was added to a stirred
suspension of T (see J. Heterocyclic. Chem. 1982, 19,673; 6.3 g,
25.12 mmol), Pd(PPh.sub.3).sub.2Cl.sub.2 (1.7 g), CuI (0.95 g) in
triethyamine (20 mL) and THF (50 mL) at room temperature and the
solution was heated at 45.degree. C. for 2 h. The solid was
filtered off and the filtration was concentrated to give dark brown
liquid. The crude product was purified by Biotage Horizon.TM.
eluting with 10%-50% EtOAc/hexane to afford title compound U (4.1
g, 60%) as a yellow solid. .sup.1H NMR (DMSO-d.sub.6, 500 MHz)
.delta. 8.11(s, 1H), 6.77(brs, 2H), 7.24(m, 1H); FIA MS
270(M+1).
2-Bromo-5H-pyrrolo[2,3-b]pyrazine (V)
[0128] A solution of U (4.0 g, 14.81 mmol) in THF (25 mL) was added
to a stirred suspension of potassium tert butoxide (2.5 g, 22.2
mmol) in THF (50 mL). The solution was refluxed for 3 h and then
stirred at room temperature for 16 h. The solvent was evaporated
and the residue was dissolved in water (100 mL) and aqueous layer
was extracted with ethyl acetate (5.times.50 mL). Organic layer was
dried and concentrated to give title compound V (1.62 g) as a
yellow solid. .sup.1H NMR (DMSO-d6, 500 MHz) .delta. 12.33(s, 1H),
8.35(s, 1H), 7.96(t, 1H), 6.63(dd, 1H); FIA MS 198(M-1).
5H-Pyrrolo[2,3-b]pyrazine (W)
[0129] 10% Palladium on carbon (1.4 g) was added to a stirred,
nitrogen flushed solution of V (1.4 g, 7.14 mmol) and ammonium
formate (4.2 g) in ethanol (50 mL). The solution was refluxed for 1
h and cooled to room temperature. The filtrate was concentrated to
give a solid, which was washed with ethyl acetate (3.times.20 mL).
The solvent was evaporated and the crude product was purified by
Biotage Horizon eluting with 40%-80% EtOAC/hexane to afford title
compound W (0.313 g, 37%) as a white solid. .sup.1H NMR (DMSO-d6,
500 MHz) .delta. 12.05(s, 1H), 8.37(d, 1H), 8.22(d, 1H), 7.85(d,
1H), 6.62(d, 1H); FIA MS 120(M+1).
7-Iodo-5H-pyrrolo[2,3-b]pyrazine (X)
[0130] 1M Solution iodine monochloride (5.26 mL, 5.26 mmol) was
added to a stirred solution of W (0.313 g, 2.63 mmol) in
CH.sub.2Cl.sub.2 (25 mL) and the solution was stirred at room
temperature for 16 h. The precipitated yellow solid was filtered
and suspended in ethyl acetate (50 mL) and washed with saturated
NaHCO.sub.3 (25 mL). The organic layer was dried and concentrated
to give title compound 22 (0.64 g, 100%). .sup.1H NMR (DMSO-d6, 500
MHz) .delta. 12.48(s, 1H), 8.45(d, 1H), 8.28(d, 1H), 8.10(d, 1H),
7.85(d, 1H), FIA MS 246(M+1).
7-Iodo-5-(toluene-4-sulfonyl)-5H-pyrrolo[2,3-b]pyrazine (Y)
[0131] Sodium hydride (0.04 g, 1.68 mmol) was added to a stirred
solution of X (0.103 g, 0.42 mmol) in THF (10 mL). The solution was
stirred at room temperature for 30 min and P-TsCl (0.81 g, 0.62
mmol) was added. The resulting suspension was heated a 60.degree.
C. for 45 min. and poured into water (10 mL). The solution was
extracted with ethyl acetate (2.times.25 mL), dried and
concentrated to give title compound Y (0.168, 85%). FIA MS
400(M+1).
2-(3-(3-Piperidin-4-yl-propoxy)-phenyl)-N-[4-(5H-pyrralo[2,3-b]pyrazin-3-
-yl)-thiophen-2-yl]-acetamide (Compound 19).
[0132] A mixture of Y (0.044 g, 0.11 mmol), H (0.055 g, 0.11 mmol),
Pd.sub.2(dba).sub.3 (0.017 g, 0.02 mmol), P(Bu.sup.t).sub.3 (0.01
g, 0.04 mmol), KF.H.sub.2O (0.1 g, 1.3 mmol) in 1,4-dioxane (2 mL)
and water (0.5 mL) was heated at 185.degree. C. in a microwave oven
for 25 min. Solid was filtered and washed with EtOAc (1 mL). The
filtrate was concentrated. The crude product was dissolved in
CH.sub.2Cl.sub.2 (1 mL) and TFA (1 mL). The solution was stirred at
room temperature for 1 h and concentrated. The crude product was
purified by preparative HPLC (5-75% CH.sub.3CN/water, 15 min) to
give title compound 19 (0.0062 g) as an oil.
2-(3-Methanesulfonylamino-phenyl)-N-[5H-pyrralo[2,3-b]pyrazin-3-yl)-thio-
phen-2-yl]-acetamide (Compound 20).
[0133] The title compound 20 was synthesized and purified in a
manner similar to 6 using a mixture of Y (0.050 g, 0.125 mmol), E
(0.044 g, 0.125 mmol), Pd.sub.2(dba).sub.3 (0.017 g, 0.02 mmol),
P(Bu.sup.t).sub.3 (0.01 g, 0.04 mmol), KF.H.sub.2O (0.1 g, 1.3
mmol) in 1,4-dioxane (2 mL) and water (0.5 mL).
Example 4
Preparation of
N-(4-(1H-pyrazolo[3,4-b]pyridin-3-yl)thiazol-2-yl)-2-phenylacetamides
[0134] ##STR50## ##STR51## t-Butyl
2-(2-fluoropyridin-3-yl)-2-oxoacetate (BB)
[0135] To a solution of 2-fluoropyridine (5.82 g, 0.0599 mol) in
150 mL THF at -78.degree. C., LDA (33.0 ml of 2M, 0.066 mol) was
added dropwise and the reaction mixture was stirred at -78.degree.
C. for 2 h. This solution was transferred slowly into chilled to
-78.degree. C. THF (500 ml) solution of t-butyl .alpha.-oxo-1
H-imidazole-1-acetate (AA) (see J. Org. Chem. 46:211, 1981; 14.69
g, 0.0749 mol) by means of cannula under nitrogen pressure. The
combined solution was continuously stirred at -78.degree. C. for 30
min. The reaction mixture was poured into sat. NH.sub.4Cl (500 mL).
The organic phase was separated and dried (MgSO.sub.4). After
removing solvent, the product was purified by chromatography to
give title compound BB (6.7 g, 49.6% yield). H NMR (CDCl.sub.3):
1.6(s, 9H), 7.4(dd, 1H), 8.36(dd, 1H), 8.5(dd, 1H).
tert-Butyl 2-(2-fluoropyridin-3-yl)-2-hydrazonoacetate (CC)
[0136] To a solution of t-butyl
2-(2-fluoropyridin-3-yl)-2-oxoacetate (BB) in methylene dichloride
at room temperature, titanium(IV) isopropoxide was added, followed
by hydrazine hydrate dropwise. The reaction mixture was stirred at
room temperature for 10 h, then 6 mL water was added. The reaction
mixture was continuously stirred at room temperature for overnight.
The reaction mixture was filtered and the filtration cake was
washed with methylene chloride 3 times. After removal of solvent,
the filtrate was purified by chromatography 95%/5% methylene
dichloride/methanol to give the title compound CC (5.0 g, 70%
yield). H NMR (CDCl.sub.3): 1.5(s, 9H), 6.18(s, br, 2H), 7.3(dd,
1H), 7.74(dd, 1H), 8.3(dd, 1H).
tert-Butyl 1H-pyrazolo[3,4]pyridine-3-carboxylate (DD)
[0137] To a solution of t-butyl
2-(2-fluoropyridin-3-yl)-2-hydrazonoacetate (CC) (0.58 g, 2.42
mmol) in THF, sodium hydride (0.116 g, 60% mineral oil dispersion,
2.9 mmol) was added in portions. The reaction mixture was stirred
at room temperature until no further gas was released, then warmed
to 50.degree. C. for 2 h. Ethyl acetate and brine was added to the
reaction mixture and the organic layer was separated and dried over
MgSO.sub.4. After removal of the solvent, the product was purified
by chromatography with 50%/50% ethyl acetate/hexanes to give title
compound DD (0.4 g, 75% yield). H NMR (CDCl.sub.3): 1.67(s, 9H),
7.28(dd, 1H), 8.52(dd, 1H), 8.63(dd, 1H).
1H-Pyrazolo[3,4-b]pyridine-3-carboxylic acid (EE)
[0138] A THF/CH.sub.2Cl.sub.2 solution of tert-butyl
1H-pyrazolo[3,4]pyridine-3-carboxylate (DD) (3.0 g, 13.7mmol) was
stirred at room temperature for 3 h. The solvent was removed to
give title compound EE (2.2 g, 99% yield). H NMR (DMSO): 7.38(dd,
1H), 8.45(dd, 1H), 8.63(dd, 1H).
1H-Pyrazolo[3,4-b]pyridine-3-carbonyl chloride (FF)
[0139] Thionyl chloride, chloroform and a catalytic amount of DMF
was added to a solution of 1H-pyrazolo[3,4-b]pyridine-3-carboxylic
acid (EE) (5.6 g, 34.3 mmol) and the mixture was refluxed
overnight. The reaction mixture was cooled to room temperature and
filtered. The filtration cake was washed with ether to give the
title compound FF.
[0140] 2-(1H-Pyrazolo[3,4-b]pyridine-3-carbonyl)-malonic acid
dimethyl ester (GG)
[0141] Sodium hydride was added in portions to a dimethylmelanate
(0.33 g, 2.5 mmol)/THF solution at 0.degree. C., then the reaction
mixture was refluxed for 1 h. A THF suspension of
1H-Pyrazolo[3,4-b]pyridine-3-carbonyl chloride (FF) was added. The
reaction mixture was refluxed overnight. 1N HCl and ethyl acetate
were added to the reaction mixture, the organic phase was separated
and dried (MgSO.sub.4) and the solvent was removed to give the
title compound GG.
1-(1H-Pyrazolo[3,4-b]pyridin-3-yl)-ethanone (HH)
[0142] H.sub.2SO.sub.4 (1 mL, 98%), H.sub.2O (5 ml) and acetic acid
(7.5 ml) was added to a solution of a crude reaction product
mixture of 2-(1H-pyrazolo[3,4-b]pyridine-3-carbonyl)-malonic acid
dimethyl ester (GG). The mixture was heated at 120.degree. C. for 3
h. The reaction mixture was adjusted to pH 6.5 and ethyl acetate
was used to extract the product. The product was purified by
chromatography with 50%/50% hexanes/ethyl acetate to give the title
compound (HH). H NMR (CDCl.sub.3): 2.80(s, 3H), 7.38(dd, 1H),
8.8.71(dd, 1H), 8.76(dd, 1H).
2-Bromo-1-(1H-pyrazolo[3,4-b]pyridin-3-yl)-ethanone (II)
[0143] To a solution of 1-(1H-pyrazolo[3,4-b]pyridin-3-yl)-ethanone
(HH) (0.66 g, 4.09 mol) in HBr (8 ml, 48%), bromine (0.654 g, 4.09
mmol) was added and the reaction mixture was stirred at 65.degree.
C. for 2 h. A precipitate was formed and filtered to give the HBr
salt of the title compound (II) (0.94 g, 71% yield). H NMR (DMSO):
4.93(s, 2H), 7.45(dd, 1H), 8.56(dd, 1H), 8.68(dd, 1H).
4-(1 H-Pyrazolo[3,4-b]pyridin-3-yl)-thiazol-2-ylamine (JJ)
[0144] An ethanol solution of
2-Bromo-1-(1H-pyrazolo[3,4-b]pyridin-3-yl)-ethanone HBr salt (II)
(0.94 g, 2.92 mol) and thiourea (0.446 g, 5.84 mol) was stirred at
room temperature overnight. A precipitate was formed and filtered
to give the title compound (JJ). H NMR (DMSO): 7.03(s, bd 2H),
7.37(dd, 1H), 7.52(s, 1H), 8.6(m, 2H).
Preparation of
N-(4-(1H-pyrazolo[3,4-b]pyridin-3-yl)thiazol-2-yl)-2-phenylacetamides
[0145] A suspension of
4-(1-H-pyrazolo[3,4-b]pyridin-3-yl)thiazol-2-ylamine (JJ) (0.065 g,
0.3 mmol), a phenylacetic acid optionally substituted with R (0.45
mmol), and Et.sub.3N (0.091 g, 0.9 mmol) in 2 mL THF is heated at
160.degree. C. for 750 sec in microwave. The final product KK is
purified by preparative HPLC.
Example 5
Preparation of
N-(4-(7H-pyrrolo[2,3-d]pyrimidin-5-yl)thiophen-2-yl)-2-(3-methsulfonamido-
phenyl)acetamide
[0146] ##STR52## 5-(2-(trimethylsilyl)ethynyl)pyrimidin-4-amine
(MM)
[0147] Trimethylsilylacetylene (3.4 mL) is added to a stirred
suspension of LL (J. Heterocyclic Chem. 19: 1285, 1982; J. Org.
Chem. 48: 1064, 1983; 25.12 mmol), Pd(PPh.sub.3).sub.2Cl.sub.2 (1.7
g), CuI (0.95 g) in triethyamine (20 mL) and THF (50 mL) at room
temperature and the solution is heated at 45.degree. C. for 2 h.
The solid is filtered off and the filtrate is concentrated to give
a dark brown liquid. The crude product is purified by Biotage
Horizon.TM. to afford title compound MM.
7H-pyrrolo[2,3-d]pyrimidine (NN)
[0148] A solution of NM (14.81 mmol) in THF (25 mL) is added to a
stirred suspension of potassium tert butoxide (2.5 g, 22.2 mmol) in
THF (50 mL). The solution is refluxed for 3 h and then stirred at
room temperature for 16 h. The solvent is evaporated and the
residue is dissolved in water (100 mL) and aqueous layer is
extracted with ethyl acetate (5.times.50 mL). Organic layer is
dried and concentrated to give title compound NN.
5-iodo-7H-pyrrolo[2,3-d]pyrimidine (OO)
[0149] A 1M solution of iodine monochloride (5.26 mL, 5.26 mmol) is
added to a stirred solution of NN (2.63 mmol) in CH.sub.2Cl.sub.2
(25 mL) and the solution is stirred at room temperature for 16 h.
The precipitated yellow solid is filtered and suspended in ethyl
acetate (50 mL) and washed with saturated NaHCO.sub.3 (25 mL). The
organic layer is dried and concentrated to give title compound
OO.
N-(4-(7H-pyrrolo[2,3-d]pyrimidin-5-yl)thiophen-2-yl)-2-(3-methsulfonamid-
ophenyl)acetamide (PP)
[0150] A mixture of 5-iodo-7H-pyrrolo[2,3-d]pyrimidine (OO) (1
mmol) and
5-(2-(3-methsulfonamidophenyl)acetamido)thiophen-3-yl-3-boronic
acid (E) (1.1 mmol), K.sub.2CO.sub.3 (3 mmol) and Pd(PPh.sub.3)4
(5%) in the 3 mL dioxane and 1 mL H.sub.2O is heated in microwave
to 150.degree. C. for ten minutes. The organic layer is separated
and concentrated and the title compound PP is purified by
preparative HPLC.
Example 6
Preparation of
N-(4-(1H-pyrazolo[3,4-d]pyrimidin-3-yl)thiophen-2-yl)-2-(3-methsulfonamid-
ophenyl)acetamide
[0151] ##STR53## 1H-Pyrazolo[3,4-d]pyrimidin-3-amine (RR)
[0152] 4-aminopyrimidine-5-carbonitrile (QQ) (1 mmol) in 10 mL
conc. HCl is cooled at 0.degree. C., to which 1.2 equiv of
NaNO.sub.2 in 1 mL H.sub.2O is added dropwise and reaction mixture
is stirred at 0.degree. C. for 10 min. To this reaction mixture, 5
equivalent of SnCl.sub.2 in 5 mL conc. HCl is added and reaction
mixture is stirred at room temperature overnight. 30 mL of EtOAc is
added to the reaction mixture and then washed with 1N NaOH. The
organic layer is separated and dried. The solvent is removed to
give title compound 1H-pyrazolo[3,4-d]pyrimidin-3-amine (RR).
3-iodo-1H-pyrazolo[3,4-d]pyrimidine (SS)
[0153] 1H-pyrazolo[3,4-d]pyrimidin-3-amine (RR) (1 mmol) in 3 mL of
sulfuric acid (16N) is cooled to 0.degree. C., then 1.2 mmol of
NaNO.sub.2 in 1 mL of H.sub.2O of water is added slowly at
0.degree. C. The reaction mixture is stirred at 0.degree. C. for 1
h. 4 mmol of potassium iodide in 3 mL H.sub.2O is poured all at
once. The mixture is warmed to room temperature over 1 h then
brought to pH of 7.8 using solid sodium carbonate and extracted
with dichloromethane. The organic phase is washed with saturated
solution of sodium thiosulfate, dried on MgSO.sub.4, and finally
evaporated under reduced pressure. The crude product is purified on
silica gel column chromatography to obtain the title compound
(SS).
N-(4-(1H-pyrazolo[3,4-d]pyrimidin-3-yl)thiophen-2-yl)-2-(3-methsulfonami-
dophenyl)acetamide (TT)
[0154] A mixture of 3-iodo-1H-pyrazolo[3,4-d]pyrimidine (SS) (1
mmol) and
5-(2-(3-methsulfonamidophenyl)acetamido)thiophen-3-yl-3-boronic
acid (E) (1.1 mmol), K.sub.2CO.sub.3 (3 mmol) and
Pd(PPh.sub.3).sub.4 (5%) in the 3 mL dioxane and 1 mL H.sub.2O is
heated in microwave to 150.degree. C. for ten minutes. The organic
layer is separated and the title compound
N-(4-(1H-pyrazolo[3,4-d]pyrimidin-3-yl)thiophen-2-yl)-2-(3-methsulfonamid-
ophenyl) acetamide (TT) is purified by preparative HPLC.
Example 7
Preparation of
N-(5-(7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1,3,4-thiadiazol-2-yl)-2-(3-methox-
yphenyl)acetamide
[0155] ##STR54##
5-(7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1,3,4-thiadiazol-2-amine
(VV)
[0156] A mixture of 7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (UU)
(See Nucleosides, Nucleotides & Nucleic Acids 20: 1823, 2001; 7
mmol) and thiosemicarbazide (2.34 g, 21 mmol) in trifluoroacetic
acid (25 mL) is heated in a sealed tube at 100.degree. C. for 2 hr.
The brown solution is cooled to room temperature and poured into
ice. The mixture is then basified with concentrated NH.sub.4OH and
the pale brown precipitate formed is filtered on a sintered glass
filter. The solid is washed thoroughly with water (3.times.50 mL)
and ethyl acetate (3.times.50 mL) and dried under vacuum to afford
5-(7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1,3,4-thiadiazol-2-amine
(VV).
N-(5-(7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1,3,4-thiadiazol-2-yl)-2-(3-metho-
xyphenyl)acetamide (WW)
[0157] A suspension of
5-(7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1,3,4-thiadiazol-2-amine (VV)
(0.23 mmol), 3-methoxyphenylacetic acid (0.038 g, 0.23 mmol),
triethylamine (0.1 ML) and BtSO.sub.2CH.sub.3 (0.055 g, 0.28 mmol)
in THF (3 mL) and DMF (0.3 mL) is heated in a microwave oven for 20
min. The brown solution is added to water (50 mL) and ethyl acetate
(50 mL). Brine (10 ml) is added to separate the layers. The organic
phase is separated and washed with water (2.times.50 mL). The
organic layer is concentrated to give a solid, which is placed in a
small Buchner funnel and washed with methanol (2.times.5 mL) and
ethyl acetate (2.times.5 mL). The brown solid collected is dried
under vacuum to afford the desired product
N-(5-(7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1,3,4-thiadiazol-2-yl)-2-(3-methox-
yphenyl)acetamide (WW).
Example 8
Preparation of
N-(5-(1H-pyrazolo[3,4-d]pyrimidin-3-yl)-1,3,4-thiadiazol-2-yl)-2-(3-metho-
xyphenyl)acetamide
[0158] ##STR55##
5-(1H-pyrazolo[3,4-d]pyrimidin-3-yl)-1,3,4-thiadiazol-2-amine
(YY)
[0159] A mixture of 1H-pyrazolo[3,4-d]pyrimidine-3-carbonitrile
(XX) (See Nucleosides, Nucleotides & Nucleic Acids 20: 1823,
2001; 7 mmol) and thiosemicarbazide (2.34 g, 21 mmol) in
trifluoroacetic acid (25 mL) is heated in a sealed tube at
100.degree. C. for 2 hr. The brown solution is cooled to room
temperature and poured into ice. The mixture is then basified with
concentrated NH.sub.4OH and the pale brown precipitate formed is
filtered on a sintered glass filter. The solid is washed thoroughly
with water (3.times.50 mL) and ethyl acetate (3.times.50 mL) and
dried under vacuum to afford
5-(1H-pyrazolo[3,4-d]pyrimidin-3-yl)-1,3,4-thiadiazol-2-amine
(YY).
N-(5-(1H-pyrazolo[3,4-d]pyrimidin-3-yl)-1,3,4-thiadiazol-2-yl)-2-(3-meth-
oxyphenyl)acetamide (ZZ)
[0160] Compound ZZ is synthesized in a manner similar to compound
WW using amine YY instead to give the desired compound ZZ.
Example 9
Preparation of
4-(1H-pyrazolo[3,4-b]pyridin-3-yl)-1H-pyrrole-2-carboxamides
[0161] ##STR56##
(2-chloropyridin-3-yl)1-(5-(2,2,2-trichloroacetyl)-1H-pyrrol-3-yl)methano-
ne (B')
[0162] Aluminum(III) chloride (20 mmol) and compound A' (Aldrich)
are dissolved in 10 mL dry CH.sub.2Cl.sub.2 at 0.degree. C.
2,2,2-trichloro-1-(1H-pyrrol-2-yl)ethanone (10 mmol) is added
dropwise and the reaction mixture is warmed to room temperature and
stirred for 2 hours. The reaction mixture is poured into ice water
and extracted with EtOAc. The organic layer is subsequently washed
with saturated NaHCO.sub.3 and brine. After drying and evaporation
of the organic solvent, the mixture is passed through flash column
chromatography to give title compound B' (540 mg, 15% yield).
[0163] Preparation of Compound C'
[0164] Compound B' (60 mg) is dissolved into CH.sub.3CN (2 mL), to
which RNH.sub.2 (1.2 equivalent) is added, followed by triethyl
amine (1.5 equivalent). The reaction mixture is stirred at room
temperature overnight. After the solvent is evaporated, the
reaction mixture is carried on to the next step without further
purification.
[0165] Preparation of Compound D'
[0166] Compound C' (reaction mixture) is dissolved in 2 mL EtOH, to
which 3 equivalent of hydrazine is added. The reaction mixture is
heated in the microwave at 170.degree. C. for 10 minutes. The
reaction mixture is directly applied on HPLC for purification to
give title compound D'.
[0167] It will be appreciated that a variety of compounds can be
prepared according to the general methods described above.
Example 10
Analytical Results
[0168] Table 2 below depicts exemplary LC mass spectral data
(LC/MS), retention time (RT) and .sup.1H-NMR data (NMR) for certain
compounds of the present invention, wherein compound numbers in
Table 2 correspond to the compounds depicted in Table 1 (empty
cells indicate that the test was not performed): TABLE-US-00002
TABLE 2 Cmpd # LC/MS RT NMR 1 2 336.00 3.20 DMSO d-6: 3.85(s, 2H),
7.28(m, 2H), 7.39(m, 4H), 7.68(s, 1H), 8,57(d, 1H), 8.72(dd, 1H),
12.52(s, br, 1H), 13.83(s, br,1H) 3 366.10 3.23 DMSO d-6: 3.77(s,
3H0, 3.80(s, 2H), 6.82(m, 1H), 6.93(m 2H), 7.3(m, 2H), 7.68(s, 1H),
8.56(d, 1H), 8.7(dd, 1H)12.47(s, br, 1H), 13.75(s, br, 1H) 4 471.10
3.50 DMSO d-6: 1.03(t, 6H), 3.15(q, 4H, 3.65(s, 2H), 7.20(m, 2H),
7.5(m, 1H), 7.6(m, 2H), 7.78(s, 1H), 8.48(dd, 1H), 8.72(dd, 1H) 5
429.10 2.79 DMSO d-6: 2.93(s, 3H), 3.76(s, 2H), 6.99(d, 1H),
7.04(dd, 1H), 7.15(s, 1H), 7.25(dd, 1H), 7.29(dd 1H), 7.64(s, 1H),
8.55(dd, 1H), 8.7(dd, 1H) 6 446.00 2.30 DMSO-d6: 11.4(brS, 1H),
8.52(m, 2H), 7.57(s, 1H), 7.27(s, 1H), 7.22(m, 1H), 6.72(m, 3H),
3.57(s, 2H), 3.17(s, 3H) 7 476.00 2.36 CD3OD: 8.54(m, 2H), 7.51(d,
1H), 7.33(d, 1H), 7.29(dd, 1H), 7.23(t, 1H), 6.91(m, 2H), 6.82(m,
1H), 4.01(t, 2H), 3.70(s, 1H), 3.34(m, 2H), 2.95(m, 2H), 1.96(m,
2H), 1.81(m, 2H), 1.65(m, 1H), 1.49(m, 2H), 1.34(m, 2H) 8 428.00
2.91 CD3OD: 8.52(m, 2H), 7.51(s, 1H), 7.14- 7.33(m, 6H), 3.74(s,
2H), 2.96(s, 3H). 9 447.00 2.79 DMSO-d6: 13.98(s, 1H), 11.58(s,
1H), 9.67(s, 1H), 8.68(d, 1H), 8.62(d, 1H), 7.83(s, 1H), 7.53(d,
1H), 7.15-7.20(m, 3H), 3.77(s, 1H), 2.79(s, 3H). 10 442.00 3.02
CD3OD: 8.57(d, 1H), 8.55(d, 1H), 7.80(s, 1H), 7.68(m, 2H), 7.60(t,
1H), 7.53(d, 1H), 7.34(d, 1H), 7.30(dd, 1H), 3.88(s, 1H), 2.69(s,
6H) 11 411.00 2.32 DMSO: 11.80 s (1H), 9.72 s (1H), 8.71 t (1H),
8.50 m (2H), 7.53 m (1H), 7.49 m (1H), 7.30 t (1H), 7.20 m (2H),
7.10 m (2H), 4.45 d (2H), 2.97 s (3H) 12 286.00 1.71 MeOD: 8.72 d
(1H), 8.60 d (1H), 7.60 s (1H), 7.42 s (1H), 7.34 m (1H), 4.18 m
(1H), 3.59 m (2H), 1.26 d (3H) 13 348.00 2.64 MeOD: 8.58 m (2H),
7.60 s (1H), 7.40 s (1H), 7.24 m (2H), 6.94 m (2H), 6.80 d
(1H),4.55 (2H), 3.78 s (3H) 14 402.10 2.60 MeOD: 8.54 m (2H), 7.60
s (1H), 7.40 m (3H), 7.28 m (2H), 7.15 d (1H), 4.64 s (2H) 15
362.10 2.59 MeOD: 8.60 m (2H), 7.60 s (1H), 7.38 s (1H), 7.29 dd
(1H), 6.87 m (2H), 6.76 m (1H), 5.91 s (2H), 4.47 s (2H) 16 335.90
2.71 MeOD: 8.56 m (2H), 7.60 s (1H), 7.39 m (3H), 7.30 m (1H), 7.05
t (2H), 4.55 s (2H) 17 324.00 2.95 MeOD: 8.55 m (2H), 7.56 s (1H),
7.36 s (1H), 7.27 m (1H), 3.20 d (2H), 1.78 m (4H), 1.67 m (2H),
1.26 m (3H), 1.0 m (2H) 18 336.00 2.70 MeOD: 8.55 m (2H), 7.60 s
(1H), 7.40 s (1H), 7.34 m (1H), 7.29 m (1H), 7.18 d (1H), 7.11 d
(1H), 6.97 m (1H), 4.58 s (2H) 19 476.00 1.70 CD3OD: 8.44(d, 1H),
8.25 (d, 1H), 7.97(s, 1H), 7.52(s, 1H), 7.33(s, 1H), 7.23(t, 1H),
6.91(m, 2H), 6.81(dd,1H), 4.01(t, 2H), 3.68(s, 2H), 3.36(m, 2H),
2.94(m, 2H), 1.93(m, 2H), 1.80(m, 2H), 1.68(brm, 1H), 1.49(m, 2H),
1.36(m, 2H). 20 428.00 2.78 CD3OD: 8.44(d, 1H), 8.29(d, 1H),
8.01(s, 1H), 7.31(m, 2H), 7.26(s, 1H), 7.17(d, 1H), 7.14(d, 1H),
3.72(s, 2H), 2.96(s, 3H)
Example 11
ROCK Inhibition Assay
[0169] Compounds are screened for their ability to inhibit ROCK I
(AA 6-553) activity using a standard coupled enzyme system (Fox et
al. Protein Sci. 7: 2249, 1998). Reactions are carried out in a
solution containing 100 mM HEPES (pH 7.5), 10 mM MgCl.sub.2, 25 mM
NaCl, 2 mM DTT and 1.5% DMSO. Final substrate concentrations in the
assay are 45 .mu.M ATP (Sigma Chemicals, St Louis, Mo.) and 200
.mu.M peptide (American Peptide, Sunnyvale, Calif.). Reactions are
carried out at 30.degree. C. and 45 nM ROCK I. Final concentrations
of the components of the coupled enzyme system are 2.5 mM
phosphoenolpyruvate, 350 .mu.M NADH, 30 .mu.g/ml pyruvate kinase
and 10 .mu.g/ml lactate dehydrogenase.
[0170] Compounds are screened for their ability to inhibit ROCK
using a standard radioactive enzyme assay. Assays are carried out
in a solution containing 100 mM HEPES (pH 7.5), 10 mM MgCl.sub.2,
25 mM NaCl, 2 mM DTT and 1.5% DMSO. Final substrate concentrations
in the assay are 13 .mu.M [.gamma.-.sup.33P] ATP (25 mCi .sup.33P
ATP/mmol ATP, Perkin Elmer, Cambridge, Mass./Sigma Chemicals, St
Louis, Mo.) and 27 .mu.M Myelin Basic Protein (MBP). Final enzyme
concentration in the assay is 5 nM ROCK. Assays are carried out at
room temperature. 1.5 .mu.l of DMSO stock containing serial
dilutions of the compound of the present invention (concentrations
ranging from 10 .mu.M to 2.6 nM) is placed in a 96 well plate. 50
.mu.l of Solution 1 (100 mM HEPES (pH 7.5), 10 mM MgCl.sub.2, 26 mM
[.gamma.-.sup.33P] ATP) is added to the plate. The reaction is
initiated by addition of 50 .mu.l of Solution 2 (100 mM HEPES (pH
7.5), 10 mM MgCl.sub.2, 4 mM DTT, 54 mM MBP and 10 nM ROCK). After
2 hours the reaction is quenched with 50 .mu.L of 30%
trichloroacetic acid (TCA, Fisher) containing 9 mM ATP. Transfer of
140 .mu.L of the quenched reaction to a glass fiber filter plate
(Corning, Cat. No. 3511) is followed by washing 3 times with 5%
TCA. 50 .mu.L of Optima Gold scintillation fluid (Perkin Elmer) is
added and the plates are counted on a Top Count (Perkin Elmer).
After removing mean background values for all of the data points
the data is fit using Prism software to obtain a K.sub.i(app).
[0171] Table 3 depicts enzyme inhibition data (K.sub.i) for certain
exemplary compounds. Compound numbers in Table 3 correspond to
those compounds depicted in Table 1.
[0172] In Table 3, "A" represents a K.sub.i of less than 0.5 .mu.M
and "B" represents a K.sub.i of between 0.5 and 5.0 .mu.M. the term
"Enzyme" indicates that an enzyme-linked assay was used; the term
".sup.33P" indicates that a radioactive assay was used.
TABLE-US-00003 TABLE 3 Cmpd # ROCK .sup.33P ROCK Enzyme 1 A 2 A 3 A
4 A 5 A 6 A 7 A 8 A 9 A 10 A 11 A 12 B 13 A 14 A 15 A 16 A 17 A 18
A 19 A 20 A
* * * * *
References